Usp 32

USP 32
Acarbose Acebutolol Hydrochloride Acebutolol Hydrochloride Capsules Acepromazine Maleate Acepromazine Maleate Injection Acepromazine Maleate Tablets Acetaminophen Acetaminophen Capsules Acetaminophen Oral Solution Acetaminophen for Effervescent Oral Solution Acetaminophen Suppositories Acetaminophen Oral Suspension Acetaminophen Tablets Acetaminophen Extended-Release Tablets Acetaminophen and Aspirin Tablets Acetaminophen, Aspirin, and Caffeine Tablets Acetaminophen and Caffeine Tablets Capsules Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine Oral Solution Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine Tablets Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine Capsules Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine Oral Powder Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine Oral Solution Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine Tablets Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine Acetaminophen, Chlorpheniramine Maleate, and Dextromethorphan Hydrobromide Tablets Acetaminophen and Codeine Phosphate Capsules Acetaminophen and Codeine Phosphate Oral Solution Acetaminophen and Codeine Phosphate Oral Suspension Acetaminophen and Codeine Phosphate Tablets Acetaminophen, Dextromethorphan Hydrobromide, Doxylamine Succinate, and Pseudoephedrine Hydrochloride Oral Solution Acetaminophen and Diphenhydramine Citrate Tablets Acetaminophen, Diphenhydramine Hydrochloride, and Pseudoephedrine Hydrochloride Tablets
MONOGRAPH LIST /
Acetaminophen and Pseudoephedrine Hydrochloride Tablets Acetazolamide Acetazolamide for Injection Acetazolamide Oral Suspension Acetazolamide Tablets Glacial Acetic Acid Acetic Acid Irrigation Acetic Acid Otic Solution Acetohexamide Acetohexamide Tablets Acetohydroxamic Acid Acetohydroxamic Acid Tablets Acetylcholine Chloride Acetylcholine Chloride for Ophthalmic Solution Acetylcysteine Acetylcysteine Solution Acetylcysteine and Isoproterenol Hydrochloride Inhalation Solution Acitretin Acitretin Capsules Acyclovir Acyclovir Capsules Acyclovir for Injection Acyclovir Ointment Acyclovir Oral Suspension Acyclovir Tablets Adenine Adenosine Adenosine Injection Medical Air Alanine Albendazole Albendazole Oral Suspension Albendazole Tablets Albumin Human Albuterol Albuterol Sulfate Albuterol Tablets Alclometasone Dipropionate Alclometasone Dipropionate Cream Alclometasone Dipropionate Ointment Alcohol Dehydrated Alcohol
Copyright 2008 The United States Pharmacopeial Convention. All Rights Reserved. For Discussion Purposes Only Not for Dissemination
/ MONOGRAPH LIST
Aluminum Chloride Aluminum Chlorohydrate Aluminum Chlorohydrate Solution Aluminum Chlorohydrex Polyethylene Glycol Aluminum Chlorohydrex Propylene Glycol Aluminum Dichlorohydrate Aluminum Dichlorohydrate Solution Aluminum Dichlorohydrex Polyethylene Glycol Aluminum Dichlorohydrex Propylene Glycol Aluminum Hydroxide Gel Dried Aluminum Hydroxide Gel Dried Aluminum Hydroxide Gel Capsules Dried Aluminum Hydroxide Gel Tablets Aluminum Phosphate Gel Aluminum Sesquichlorohydrate Aluminum Sesquichlorohydrate Solution Aluminum Sesquichlorohydrex Polyethylene Glycol Aluminum Sesquichlorohydrex Propylene Glycol Aluminum Subacetate Topical Solution Aluminum Sulfate
USP 32
Dehydrated Alcohol Injection Rubbing Alcohol Alcohol in Dextrose Injection Alendronate Sodium Alendronate Sodium Tablets Alfentanil Hydrochloride Alfentanil Injection Alfuzosin Hydrochloride Allantoin Allopurinol Allopurinol Oral Suspension Allopurinol Tablets Allyl Isothiocyanate Aloe Alprazolam Alprazolam Oral Suspension Alprazolam Tablets Alprostadil Alprostadil Injection Alteplase Alteplase for Injection Altretamine Altretamine Capsules Ammonium Alum Potassium Alum Alumina and Magnesia Oral Suspension Alumina and Magnesia Tablets Alumina, Magnesia, and Calcium Carbonate Oral Suspension Alumina, Magnesia, and Calcium Carbonate Tablets Alumina, Magnesia, and Calcium Carbonate Chewable Tablets Alumina, Magnesia, Calcium Carbonate, and Simethicone Tablets Alumina, Magnesia, Calcium Carbonate, and Simethicone Chewable Tablets Alumina, Magnesia, and Simethicone Oral Suspension Alumina, Magnesia, and Simethicone Tablets Alumina, Magnesia, and Simethicone Chewable Tablets Alumina and Magnesium Carbonate Oral Suspension Alumina and Magnesium Carbonate Tablets Alumina, Magnesium Carbonate, and Magnesium Oxide Tablets Alumina and Magnesium Trisilicate Oral Suspension Alumina and Magnesium Trisilicate Tablets Aluminum Acetate Topical Solution
Aluminum Sulfate and Calcium Acetate for Topical Solution Aluminum Sulfate and Calcium Acetate Tablets for Topical Solution Aluminum Zirconium Octachlorohydrate Aluminum Zirconium Octachlorohydrate Solution Aluminum Zirconium Octachlorohydrex Gly Aluminum Zirconium Octachlorohydrex Gly Solution Aluminum Zirconium Pentachlorohydrate Aluminum Zirconium Pentachlorohydrate Solution Aluminum Zirconium Pentachlorohydrex Gly Aluminum Zirconium Pentachlorohydrex Gly Solution Aluminum Zirconium Tetrachlorohydrate Aluminum Zirconium Tetrachlorohydrate Solution Aluminum Zirconium Tetrachlorohydrex Gly Aluminum Zirconium Tetrachlorohydrex Gly Solution Aluminum Zirconium Trichlorohydrate Aluminum Zirconium Trichlorohydrate Solution Aluminum Zirconium Trichlorohydrex Gly Aluminum Zirconium Trichlorohydrex Gly Solution Amantadine Hydrochloride Amantadine Hydrochloride Capsules Amantadine Hydrochloride Oral Solution Amcinonide
USP 32
Amcinonide Cream Amcinonide Ointment Amifostine Amifostine for Injection Amikacin Amikacin Sulfate Amikacin Sulfate Injection Amiloride Hydrochloride Amiloride Hydrochloride Tablets Amiloride Hydrochloride and Hydrochlorothiazide Tablets Amiloxate Aminobenzoate Potassium Aminobenzoate Potassium Capsules Aminobenzoate Potassium for Oral Solution Aminobenzoate Potassium Tablets Aminobenzoate Sodium Aminobenzoic Acid Aminobenzoic Acid Gel Aminobenzoic Acid Topical Solution Aminocaproic Acid Aminocaproic Acid Injection Aminocaproic Acid Oral Solution Aminocaproic Acid Tablets Aminoglutethimide Aminoglutethimide Tablets Aminohippurate Sodium Injection Aminohippuric Acid Aminopentamide Sulfate Aminopentamide Sulfate Injection Aminopentamide Sulfate Tablets Aminophylline Aminophylline Injection Aminophylline Oral Solution Aminophylline Rectal Solution Aminophylline Suppositories Aminophylline Tablets Aminophylline Delayed-Release Tablets Aminosalicylate Sodium Aminosalicylate Sodium Tablets Aminosalicylic Acid Aminosalicylic Acid Tablets Amitraz Amitraz Concentrate for Dip Amitriptyline Hydrochloride
MONOGRAPH LIST /
Amitriptyline Hydrochloride Injection Amitriptyline Hydrochloride Tablets Amlodipine Besylate Aromatic Ammonia Spirit Ammonium Chloride Ammonium Chloride Injection Ammonium Chloride Delayed-Release Tablets Ferric Ammonium Citrate Ferric Ammonium Citrate for Oral Solution Ammonium Molybdate Ammonium Molybdate Injection Amobarbital Sodium Amobarbital Sodium for Injection Amodiaquine Amodiaquine Hydrochloride Amodiaquine Hydrochloride Tablets Amoxapine Amoxapine Tablets Amoxicillin Amoxicillin Boluses Amoxicillin Capsules Amoxicillin Intramammary Infusion Amoxicillin for Injectable Suspension Amoxicillin Oral Suspension Amoxicillin for Oral Suspension Amoxicillin Tablets Amoxicillin Tablets for Oral Suspension Amoxicillin and Clavulanate Potassium for Oral Suspension Amoxicillin and Clavulanate Potassium Tablets Amphetamine Sulfate Amphetamine Sulfate Tablets Amphotericin B Amphotericin B Cream Amphotericin B for Injection Amphotericin B Lotion Amphotericin B Ointment Ampicillin Ampicillin Boluses Ampicillin Capsules Ampicillin for Injection Ampicillin Soluble Powder Ampicillin for Injectable Suspension
/ MONOGRAPH LIST
Arginine Hydrochloride Arginine Hydrochloride Injection Arsanilic Acid Ascorbic Acid Ascorbic Acid Injection Ascorbic Acid Oral Solution Ascorbic Acid Tablets Aspartic Acid Aspirin Aspirin Boluses Aspirin Capsules Aspirin Delayed-Release Capsules Aspirin Suppositories Aspirin Tablets Buffered Aspirin Tablets Aspirin Delayed-Release Tablets Aspirin Effervescent Tablets for Oral Solution Aspirin Extended-Release Tablets Aspirin, Alumina, and Magnesia Tablets Aspirin, Alumina, and Magnesium Oxide Tablets
USP 32
Ampicillin for Oral Suspension Ampicillin Tablets Ampicillin and Probenecid for Oral Suspension Ampicillin Sodium Ampicillin and Sulbactam for Injection Amprolium Amprolium Soluble Powder Amprolium Oral Solution Amyl Nitrite Amyl Nitrite Inhalant Anileridine Anileridine Injection Anileridine Hydrochloride Anileridine Hydrochloride Tablets Antazoline Phosphate Anthralin Anthralin Cream Anthralin Ointment Anthrax Vaccine Adsorbed Anticoagulant Citrate Dextrose Solution Anticoagulant Citrate Phosphate Dextrose Solution Anticoagulant Citrate Phosphate Dextrose Adenine Solution Anticoagulant Heparin Solution Anticoagulant Sodium Citrate Solution Antihemophilic Factor Cryoprecipitated Antihemophilic Factor Antimony Potassium Tartrate Antimony Sodium Tartrate Antipyrine Antipyrine and Benzocaine Otic Solution Antipyrine, Benzocaine, and Phenylephrine Hydrochloride Otic Solution Antithrombin III Human Antivenin (Crotalidae) Polyvalent Antivenin (Latrodectus mactans) Antivenin (Micrurus fulvius) Apomorphine Hydrochloride Apomorphine Hydrochloride Tablets Apraclonidine Hydrochloride Apraclonidine Ophthalmic Solution Aprotinin Aprotinin Injection Arginine
Aspirin, Caffeine, and Dihydrocodeine Bitartrate Capsules Aspirin and Codeine Phosphate Tablets Aspirin, Codeine Phosphate, Alumina, and Magnesia Tablets Astemizole Astemizole Tablets Atenolol Atenolol Injection Atenolol Oral Solution Atenolol Tablets Atenolol and Chlorthalidone Tablets Atovaquone Atovaquone Oral Suspension Atracurium Besylate Atracurium Besylate Injection Atropine Atropine Sulfate Atropine Sulfate Injection Atropine Sulfate Ophthalmic Ointment Atropine Sulfate Ophthalmic Solution Atropine Sulfate Tablets Activated Attapulgite Colloidal Activated Attapulgite Aurothioglucose
USP 32
Aurothioglucose Injectable Suspension Avobenzone Azaperone Azaperone Injection Azatadine Maleate Azatadine Maleate Tablets Azathioprine Azathioprine Oral Suspension Azathioprine Tablets
MONOGRAPH LIST /
Azathioprine Sodium for Injection Azithromycin Azithromycin Capsules Azithromycin for Oral Suspension Aztreonam Aztreonam Injection Aztreonam for Injection
USP 32
Official Monographs / Acarbose 1

Analysis Samples: Standard solution and Sample solution Calculate the percentage of C25H43NO18 in the portion of Acarbose taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acarbose RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 95.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2%, determined on 1.0 g HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Sample stock solution: Use the Sample solution, as directed in the Assay. Sample solution: Sample stock solution and water (1:99) Analysis Samples: Sample stock solution and Sample solution Calculate the percentage of each impurity in the portion of Acarbose taken: Result = (ri/ra) RRF 100 = peak response for each impurity = response of the main acarbose peak in the chromatogram of the Sample solution RRF = relative response factor for each impurity, as listed in the Impurity Table Acceptance criteria Individual impurities: See Impurity Table. Total impurities: NMT 3.0% ri ra
Impurity Table Relative Retention Time 0.9 0.8 1.2 0.5 1.7 1.9 2.2 0.6 Relative Response Factor 1 1.6 1 1.33 0.8 0.8 0.8 1 Acceptance Criteria, NMT (%) 0.6 0.5 1.5 1.0 0.2 0.3 0.3 0.2
Acarbose
132270
(Comment on this Monograph)id=m115=Acarbose=AMonos.pdf)
rU rS CS
C25H43NO18 645.60 D-Glucose, O-4,6-dideoxy-4-[[[1S-(1,4,5,6)]-4,5,6trihydroxy-3-(hydroxymethyl)-2-cyclohexen-1-yl]amino]--Dglucopyranosyl-(14)-O--D-glucopyranosyl-(14)-; O-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3(hydroxymethyl)-2-cyclohexen-1-yl]amino]--Dglucopyranosyl-(14)-O--D-glucopyranosyl-(14)-Dglucose [56180-94-0]. DEFINITION Acarbose is produced by certain strains of Actinoplanes utahensis. It contains NLT 95.0% and NMT 102.0% of C25H43NO18, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.6 mg/mL of monobasic potassium phosphate and 0.35 mg/mL of dibasic sodium phosphate Mobile phase: Acetonitrile and Solution A (3:1) System suitability solution: Reconstitute a vial of USP Acarbose System Suitability Mixture RS in 1 mL of water. Standard solution: Reconstitute a vial of USP Acarbose RS in 5.0 mL of water. Sample solution: 20 mg/mL of Acarbose in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4-mm 25-cm; packing L8 Temperature: 35 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEIdentify the acarbose peak and the peaks due to the impurities listed in the Impurity Table.] Suitability requirements Height to valley ratio: NLT 1.2 for the impurity A peak, and the valley between the impurity A peak and the acarbose peak [NOTEThe chromatogram obtained is similar to the chromatogram supplied with USP Acarbose System Suitability Mixture RS.]
Name Impurity Aa Impurity Bb Impurity Cc Impurity Dd Impurity Ee Impurity Ff Impurity Gg Impurity Hh
Acarbose / Official Monographs

Impurity Table (continued) Relative Retention Time Relative Response Factor Acceptance Criteria, NMT (%) 0.2
USP 32
USP Acarbose System Suitability Mixture RS
Name Any individual unknown impurity
Acebutolol Hydrochloride
(Comment on this Monograph)id=m125=Acebutolol Hydrochloride=A-Monos.pdf)
aO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-
(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--Dglucopyranosyl-(14)-D-arabino-hex-2-ulopyranose. b(1R,4R,5S,6R)-4,5,6-Trihydroxy-2-(hydroxymethyl)cyclohex-2-enyl 4-O[4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl]--Dglucopyranoside. c-D-Glucopyranosyl 4-O-[4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl]-D-glucopyranoside. d4-O-[4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl]-Dglucopyranose. eO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--Dglucopyranosyl-(14)-O--D-glucopyranosyl-(14)-D-arabino-hex-2ulopyranose (4-O--acarbosyl-D-fructopyranose). fO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--Dglucopyranosyl-(14)-O--D-glucopyranosyl-(14)-D-glucopyranose (4-O-acarbosyl-D-glucopyranose)O-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl(14)-O--D-glucopyranosyl-(14)-O--D-glucopyranosyl-(14)-Dglucopyranose (4-O--acarbosyl-D-glucopyranose). g-D-Glucopyranosyl O-4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O--Dglucopyranosyl-(14)-O--D-glucopyranoside (-D-glucopyranosyl acarboside)-D-Glucopyranosyl O-4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl(14)-O--D-glucopyranosyl-(14)-O--D-glucopyranoside (-Dglucopyranosyl -acarboside). hO-4,6-Dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3(hydroxymethyl)cyclohex-2-enyl]amino]--D-glucopyranosyl-(14)-O-6deoxy--D-glucopyranosyl-(14)-D-glucopyranoseO-4,6-Dideoxy-4-[ [(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl]amino]-D-glucopyranosyl-(14)-O-6-deoxy--D-glucopyranosyl-(14)-Dglucopyranose.
C18H28N2O4 HCl 372.89 Butanamide, N-[3-acetyl-4-[2-hydroxy-3-[(1methylethyl)amino]propoxy]phenyl]-, monohydrochloride, ()-; ()-3-Acetyl-4-[2-hydroxy-3-(isopropylamino)propoxy]butyranilide monohydrochloride [34381-68-5]. DEFINITION Acebutolol Hydrochloride contains NLT 98.0% and NMT 102.0% of C18H28N2O4 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solution: Prepare a mixture of the Standard solution and the Sample solution (1:1) from the Assay. Analysis: Chromatograph the Sample solution as directed in the Assay. Acceptance criteria: The chromatogram thus obtained exhibits a single major peak due to acebutolol. C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements, when tested as directed for alkaloidal hydrochlorides ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and 0.3% aqueous solution of sodium dodecyl sulfate (675:20:325) [NOTEMake adjustments if necessary to achieve a retention time for acebutolol of between 4 and 7 min.] Standard solution: 0.14 mg/mL of USP Acebutolol Hydrochloride RS Sample solution: 0.14 mg/mL of Acebutolol Hydrochloride Chromatographic system (See Chromatography 621, System Suitability.)
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +168 to +183 Sample solution: 10 mg/mL PH 791: 5.57.5, in a solution containing 50 mg/mL WATER DETERMINATION, Method Ic 921: NMT 4.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Acarbose RS
USP 32
Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H28N2O4 HCl in the Acebutolol Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acebutolol Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Standard solution: 1.0 mg/mL of USP Acebutolol Hydrochloride RS in methanol Reference solution A: Dilute 3.0 mL of the Standard solution with methanol to 100 mL. Reference solution B: Dilute 5.0 mL of the Standard solution with methanol to 100 mL. Sample solution A: 10 mg/mL of Acebutolol Hydrochloride in methanol Sample solution B: Sample solution A and methanol (1:9) Developing solvent system: Butyl alcohol, glacial acetic acid, and water (4:1:5) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Analysis Samples: Standard solution, Reference solution A, Reference solution B, Sample solution A, and Sample solution B Proceed as directed in the chapter. Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths the length of the plate. Examine the plate under short-wavelength UV light. Acceptance criteria: The chromatograms from Sample solution B and the Standard solution show principal spots at the same RF value. No secondary spot from Sample solution A, excluding the area at the point of application, is more intense than the principal spot of Reference solution A (0.3%); NMT two secondary spots from Sample solution A are more intense than the principal spot of Reference solution B (0.1%); and the total of all impurities detected in the chromatogram of Sample solution A is NMT 0.5%. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 140144 PH 791: 4.57.0, in a solution (1 in 100) LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight
Official Monographs / Acebutolol 3

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acebutolol Hydrochloride RS
Acebutolol Hydrochloride Capsules

(Comment on this Monograph)id=m127=Acebutolol Hydrochloride Capsules=A-Monos.pdf) DEFINITION Acebutolol Hydrochloride Capsules contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of acebutolol (C18H28N2O4). IDENTIFICATION The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Dissolve 2.4 g of sodium 1-decanesulfonate in 1000 mL of water. Adjust with glacial acetic acid to a pH of 3.5. Mobile phase: Methanol and Solution A (3:2) Standard solution: 0.22 mg/mL of USP Acebutolol Hydrochloride RS in methanol [NOTEThis is equivalent to 0.2 mg/mL of acebutolol.] Sample stock solution: Weigh and mix, as completely as possible, the contents of NLT 20 Capsules. Transfer a portion of the powder, equivalent to 200 mg of acebutolol, to a 200-mL volumetric flask. Add 180 mL of methanol, and stir by mechanical means for 30 min. Dilute with methanol to volume. Sample solution: Dilute 5.0 mL of the Sample stock solution with methanol to 25 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H28N2O4 in the Capsules taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 Mr2 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acebutolol Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL) = molecular weight of acebutolol, 336.44 = molecular weight of acebutolol hydrochloride, 372.89
Acebutolol / Official Monographs

Acceptance criteria: 90.0%110.0% rS CS CU Mr1 Mr2
USP 32
= peak response from the Standard solution = concentration of USP Acebutolol Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of the Sample solution (g/mL) = molecular weight of acebutolol, 336.44 = molecular weight of acebutolol hydrochloride, 372.89
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Standard solution: USP Acebutolol Hydrochloride RS in Medium Spectrometric conditions Mode: UV Analytical wavelength: 232 nm Analysis: Determine the amount of C18H28N2O4 dissolved by employing the UV absorption on filtered portions of the Sample solution in comparison with a Standard solution in the same Medium. Tolerances: NLT 80% (Q) of the labeled amount of C18H28N2O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Test 1 Solution A: Prepare as directed in the Assay. Mobile phase: Methanol and Solution A (44:56) Diluent: Methanol and Solution A (1:1) Standard solution: Transfer 30 mg of USP Acebutolol Hydrochloride RS to a 50-mL volumetric flask. Add 12 mL of methanol, swirl to dissolve, and dilute with Diluent to volume. Dilute a volume of this solution with Diluent to obtain a concentration of 1.4 g/mL of USP Acebutolol Hydrochloride RS. Sample solution: Transfer a portion of the contents of 20 opened Capsules, equivalent to 250 mg of acebutolol, to a 100-mL volumetric flask, add 25 mL of methanol, and shake by mechanical means for 15 min. Dilute with Diluent to volume. Centrifuge a portion of this solution, and transfer 10.0 mL of the clear supernatant to a 100-mL volumetric flask. Dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 3.9-mm 15-cm; 4-m packing L1 Flow rate: 1 mL/min Injection size: 35 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 6.0% Analysis Samples: Diluent, Standard solution, and Sample solution [NOTERecord the chromatograms for two times the retention time of acebutolol, and measure the responses for all the peaks, disregarding any peaks corresponding to those obtained from the Diluent.] Calculate the percentage of each impurity eluting prior to the acebutolol peak in the portion of Capsules taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU = peak response for any individual impurity from the Sample solution
Test 2 Solution A: Prepare as directed in the Assay. Mobile phase: Methanol and Solution A (1:1) Standard solution: Transfer 30 mg of USP Acebutolol Hydrochloride RS to a 50-mL volumetric flask. Add 12 mL of methanol, swirl to dissolve, and dilute with Mobile phase to volume. Dilute a volume of this stock solution with Mobile phase to obtain a concentration of 1.4 g/mL of USP Acebutolol Hydrochloride RS. Sample solution: Transfer a portion of the contents of 20 opened Capsules, equivalent to 250 mg of acebutolol, to a 100-mL volumetric flask, add 25 mL of methanol, and shake by mechanical means for 15 min. Dilute with Mobile phase to volume. Centrifuge a portion of this solution, and transfer 10.0 mL of the clear supernatant to a 100-mL volumetric flask. Dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 3.9 mm 15 cm; 4-m packing L1 Flow rate: 1 mL/min Injection size: 70 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 6.0% Analysis Samples: Mobile phase, Standard solution, and Sample solution Calculate the percentage of each impurity eluting after the acebutolol peak in the portion of Capsules taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response for any individual impurity from the Sample solution = peak response from the Standard solution rS = concentration of USP Acebutolol Hydrochloride CS RS in the Standard solution (g/mL) = nominal concentration of the Sample solution CU (g/mL) = molecular weight of acebutolol, 336.44 Mr1 = molecular weight of acebutolol hydrochloride, Mr2 372.89 Acceptance criteria Test 1: NMT 0.5% of any individual impurity Test 2: NMT 0.5% of any individual impurity Sum of impurities found in Test 1 and Test 2: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acebutolol Hydrochloride RS rU
USP 32
Official Monographs / Acepromazine 5

Acceptance criteria: 98.0%101.0%
Acepromazine Maleate
(Comment on this Monograph)id=m134=Acepromazine Maleate=A-Monos.pdf)
C19H22N2OS C4H4O4 442.53 Ethanone, 1-[10-[3-(dimethylamino)propyl]-10H-phenothiazin-2yl]-, (Z)-2-butenedioate (1:1); 10-[3-(Dimethylamino)propyl]phenothiazin-2-yl methyl ketone maleate (1:1) [3598-37-6]. DEFINITION Acepromazine Maleate contains NLT 98.0% and NMT 101.0% of C19H22N2OS C4H4O4, calculated on the anhydrous basis. [NOTEThroughout the following procedures, protect samples, the USP Reference Standard, and solutions containing them, by conducting the procedures without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the peak from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Add 6 mL of triethylamine to 700 mL of water, and adjust with phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile and Solution A (300:700) Standard stock solution: 1 mg/mL of USP Acepromazine Maleate RS in 0.05 N hydrochloric acid Standard solution: 0.1 mg/mL of USP Acepromazine Maleate RS in water, from Standard stock solution Sample stock solution: 1 mg/mL of Acepromazine Maleate in 0.05 N hydrochloric acid Sample solution: 0.1 mg/mL of Acepromazine Maleate in water, from Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4 mm 15 cm; 5-m packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H22N2OS C4H4O4 in the Acepromazine Maleate taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acepromazine Maleate RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL)
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE [NOTEConduct this test without exposure to daylight, and with the minimum necessary exposure to artificial light.] Diluent: Methanol and diethylamine (19:1) Standard solution: Dilute 1 volume of the Sample solution with 200 volumes of Diluent (0.1 mg/mL). Sample solution: 20.0 mg/mL in Diluent Developing solvent system: n-Heptane, isobutyl alcohol, and diethylamine (75:17:8) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Analysis Samples: Standard solution and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths the length of the plate. Examine the plate under shortwavelength UV light. Acceptance criteria: No spot, other than the principal acepromazine spot and any at the origin, observed in the chromatogram of the Sample solution is more intense than the principal spot observed in the chromatogram of the Standard solution (0.5%). SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 136139 PH 791: 4.05.5 in a solution (1 in 100) WATER DETERMINATION, Method I 921: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, protected from light. Store at room temperature. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Acepromazine Maleate RS
Acepromazine Maleate Injection

(Comment on this Monograph)id=m137=Acepromazine Maleate Injection=A-Monos.pdf) DEFINITION Acepromazine Maleate Injection is a sterile solution of Acepromazine Maleate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of acepromazine maleate (C19H22N2OS C4H4O4). [NOTEThroughout the following procedures, protect samples, the Reference Standard, and solutions containing them, by conducting the procedures without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: To a volume of Injection equivalent to 20 mg of acepromazine maleate, add 2 mL of water and 3 mL of 2 N sodium hydroxide, and extract with two 5-mL portions of cyclohexane. Combine the cyclohexane extracts, and
Acepromazine / Official Monographs
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evaporate to dryness under vacuum, using gentle heat, if necessary. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Add 6 mL of triethylamine to 700 mL of water and adjust with phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile and Solution A (300:700) Standard stock solution: 1 mg/mL of USP Acepromazine Maleate RS in 0.05 N hydrochloric acid Standard solution: 0.1 mg/mL of USP Acepromazine Maleate RS in water, from Standard stock solution Sample stock solution: 1 mg/mL of acepromazine maleate in 0.05 N hydrochloric acid, from an appropriately diluted volume of Injection Sample solution: 0.1 mg/mL of acepromazine maleate in water, from Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4 mm 15 cm; 5-m packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H22N2OS C4H4O4 in the volume of Injection taken: Result = (rU/rS) (CS/CU) 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Acepromazine Maleate RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 4.55.8 OTHER REQUIREMENTS: It meets the requirements under Injections 1. BACTERIAL ENDOTOXINS TEST 85: NMT 4.5 USP Endotoxin Units/mg of acepromazine maleate STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant, single-dose or multiple-dose Containers for Injections as described under Injections 1, preferably of Type I glass, and store at controlled room temperature. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Acepromazine Maleate RS USP Endotoxin RS
Acepromazine Maleate Tablets

(Comment on this Monograph)id=m140=Acepromazine Maleate Tablets=A-Monos.pdf) DEFINITION Acepromazine Maleate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of acepromazine maleate (C19H22N2OS C4H4O4). [NOTEThroughout the following procedures, protect samples, the Reference Standard, and solutions containing them, by conducting the procedures without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: To a quantity of powdered Tablets equivalent to 20 mg of acepromazine maleate, add 2 mL of water and 3 mL of 2 N sodium hydroxide, and extract with two 5-mL portions of cyclohexane. Combine the cyclohexane extracts, and evaporate to dryness under vacuum, using gentle heat if necessary. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Add 6 mL of triethylamine to 700 mL of water, and adjust with phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile and Solution A (300:700) Standard stock solution: 1 mg/mL of USP Acepromazine Maleate RS in 0.05 N hydrochloric acid Standard solution: 0.1 mg/mL of USP Acepromazine Maleate RS in water, from Standard stock solution Sample stock solution: Transfer NLT 10 Tablets to a 200mL volumetric flask, add 100 mL of 0.05 N hydrochloric acid, and sonicate for 10 min. Shake by mechanical means for 30 min, and dilute with 0.05 N hydrochloric acid to volume. Sample solution: 0.1 mg/mL of acepromazine maleate in water, from Sample stock solution. Pass a portion of this solution through a filter having a 0.5-m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4 mm 15 cm; 5-m packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H22N2OS C4H4O4 in the Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Acepromazine Maleate RS in the Standard solution (mg/mL)
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= nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. LABELING: Label the Tablets to indicate that they are for veterinary use only. USP REFERENCE STANDARDS 11 USP Acepromazine Maleate RS CU
Official Monographs / Acetaminophen 7

Acetaminophen
(Comment on this Monograph)id=m150=Acetaminophen=AMonos.pdf)
C8H9NO2 Acetamide, N-(4-hydroxyphenyl)-; 4-Hydroxyacetanilide [103-90-2].
151.16
DEFINITION Acetaminophen contains NLT 98.0% and NMT 101.0% of C8H9NO2, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 5 g/mL in a mixture of 0.1 N hydrochloric acid in methanol (1 in 100) C. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 1 mg/mL in methanol Developing solvent system: Methylene chloride and methanol (4:1) Acceptance criteria: Meets the requirements of the test ASSAY PROCEDURE Sample solution: Dissolve 120 mg of Acetaminophen in 10 mL of methanol in a 500-mL volumetric flask, and dilute with water to volume. Dilute 5.0 mL of the resulting solution with water to 100 mL. Standard solution: 12 g/mL of USP Acetaminophen RS in the same medium as Sample solution Spectrometric conditions Mode: UV Analytical wavelength: 244 nm Cell: 1 cm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the portion of Acetaminophen taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (g/mL) = concentration of the Sample solution (g/mL)
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221 Sample solution: Shake 1.0 g of Acetaminophen with 25 mL of water, filter, and add 1 mL of 2 N nitric acid and 1 mL of silver nitrate TS. Acceptance criteria: The filtrate shows no more chloride than corresponds to 0.20 mL of 0.020 N hydrochloric acid (0.014%). CHLORIDE AND SULFATE, Sulfate 221 Sample solution: Shake 1.0 g of Acetaminophen with 25 mL of water, filter, and add 2 mL of 1 N acetic acid, then add 2 mL of barium chloride TS. Acceptance criteria: The mixture shows no more sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid (0.02%). SULFIDE Sample: 2.5 g Analysis: Place the Sample in a 50-mL beaker. Add 5 mL of alcohol and 1 mL of 3 N hydrochloric acid. Moisten a piece of lead acetate test paper with water, and fix to the underside of a watch glass. Cover the beaker with the watch glass so that part of the lead acetate paper hangs down near the pouring spout of the beaker. Heat the contents of the beaker on a hot plate just to boiling. Acceptance criteria: No coloration or spotting of the test paper occurs. HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE 1 Alkaline nitroferricyanide solution: Dissolve1 g of sodium nitroferricyanide and 1 g of anhydrous sodium carbonate in 100 mL of water. Sample solution: Transfer 5.0 g of Acetaminophen to a 100-mL volumetric flask, and dissolve in 75 mL of a mixture of equal volumes of methanol and water. Add 5.0 mL of Alkaline nitroferricyanide solution, dilute with a mixture of equal volumes of methanol and water to volume, and allow to stand for 30 min. Standard solution: 2.5 g/mL of p-aminophenol, similarly prepared as Sample solution, using the same quantities of the same reagents Blank: 5.0 mL of Alkaline nitroferricyanide solution, diluted in a mixture of equal volumes of methanol and water to 100 mL Spectrometric conditions Mode: UVVis Analytical wavelength: 710 nm Cell: 1 cm Analysis Samples: Sample solution, Standard solution, and Blank Acceptance criteria: The absorbance of the Sample solution does not exceed that of the Standard solution, corresponding to NMT 50 ppm of p-aminophenol. PROCEDURE 2 Standard solution: 10 g/mL of p-chloroacetanilide in ether Sample solution: Transfer 1.0 g of Acetaminophen to a glass-stoppered, 15-mL centrifuge tube, add 5.0 mL of ether, shake by mechanical means for 30 min, and centrifuge at 1000 rpm for 15 min or until a clean separation is obtained. The supernatant is used as the Sample solution.
Acetaminophen / Official Monographs

Chromatographic system (see Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: Apply 200 L of the Sample solution, in 40-L portions, to obtain a single spot NMT 10 mm in diameter. Developing solvent system: Solvent hexane and acetone (3:1) Analysis Samples: Standard solution and Sample solution Develop the chromatogram in an unsaturated chamber with the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Examine the plate under short-wavelength UV light. Acceptance criteria: Any spot in the chromatogram of the Sample solution, at an RF value corresponding to the principal spot from the Standard solution, is not greater in size or intensity than the principal spot of the Standard solution (NMT 10 ppm).
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having a 0.5-m or finer porosity, discarding the first 10 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 243 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Detector: UV 249 nm Sample solution: Sample per Dissolution 711. Standard solution: USP Acetaminophen RS in Medium Analysis: Determine the amount of C8H9NO2 dissolved by using UV absorption on filtered portions of the Sample solution suitably diluted with Medium, if necessary, in comparison with the Standard solution having a known concentration of USP Acetaminophen RS. Tolerances: NLT 75% (Q) of the labeled amount of C8H9NO2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS rU rS CS
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 168172 WATER DETERMINATION, Method I 921: NMT 0.5% READILY CARBONIZABLE SUBSTANCES TEST 271: Dissolve 0.50 g in 5 mL of sulfuric acid TS: the solution has no more color than Matching Fluid A. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at room temperature. Protect from moisture and heat. USP REFERENCE STANDARDS 11 USP Acetaminophen RS
Acetaminophen Capsules
(Comment on this Monograph)id=m160=Acetaminophen Capsules=A-Monos.pdf) DEFINITION Acetaminophen Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of acetaminophen (C8H9NO2). IDENTIFICATION A. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Equivalent to 1 mg/mL of acetaminophen from contents of the Capsules in methanol. Filter and use filtrate. Developing solvent system: Methylene chloride and methanol (4:1) Acceptance criteria: Meet the requirements ASSAY PROCEDURE Mobile phase: Methanol and water (1:3) Standard solution: 0.01 mg/mL of USP Acetaminophen RS in Mobile phase Sample solution: Weigh the contents of NLT 20 Capsules, and calculate the average weight of the contents of each Capsule. Mix the combined contents of the Capsules, and transfer a portion, equivalent to 100 mg of acetaminophen, to a 200-mL volumetric flask. Add 100 mL of Mobile phase, shake by mechanical means for 10 min, and dilute with Mobile phase to volume. Transfer 5.0 mL of this solution to a 250-mL volumetric flask and dilute with Mobile phase to volume. Pass a portion of this solution through a filter
Acetaminophen Oral Solution

(Comment on this Monograph)id=m180=Acetaminophen Oral Solution=A-Monos.pdf) DEFINITION Acetaminophen Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of acetaminophen (C8H9NO2). IDENTIFICATION A. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Equivalent to 1 mg/mL of acetaminophen in methanol Developing solvent system: Methylene chloride and methanol (4:1)
USP 32
Acceptance criteria: Meets the requirements
ASSAY PROCEDURE Mobile phase: Methanol and water (1:3) Standard solution: 0.01 mg/mL of USP Acetaminophen RS in Mobile phase Sample solution: Transfer a measured volume of Oral Solution, equivalent to 500 mg of acetaminophen, to a 250mL volumetric flask, and dilute with Mobile phase to volume. Transfer 5.0 mL of this solution to a second 250-mL volumetric flask, and dilute with Mobile phase to volume. Transfer 25.0 mL of this solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity, discarding the first 10 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 243 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) CU = nominal concentration of acetaminophen in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements for oral solution packaged in multiple-unit containers UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for oral solution packaged in single-unit containers SPECIFIC TESTS PH 791: 3.86.1 ALCOHOL DETERMINATION, Method II 611: Between 90.0% and 115.0% of the labeled amount of C2H5OH, determined by the gasliquid chromatographic procedure, with acetone being used as the internal standard ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS
Acetaminophen for Effervescent Oral Solution

(Comment on this Monograph)id=m183=Acetaminophen for Effervescent Oral Solution=A-Monos.pdf) DEFINITION Acetaminophen for Effervescent Oral Solution contains, in each 100 g, NLT 5.63 g and NMT 6.88 g of acetaminophen (C8H9NO2). IDENTIFICATION A. A 10-g portion dissolves, with effervescence, in water when prepared as directed for the Sample solution in the Assay. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Triturate 0.4 g of the powder with 25 mL of methanol, and filter. Developing solvent system: Methylene chloride and methanol (4:1) Acceptance criteria: Meets the requirements of the test ASSAY PROCEDURE Mobile phase: Methanol and water (1:3) Standard solution: 0.01 mg/mL of USP Acetaminophen RS in Mobile phase Sample solution: Dissolve 10 g of Acetaminophen for Effervescent Oral Solution in 200 mL of water in a 1000-mL volumetric flask, using gentle heat if necessary, until effervescence subsides, then dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 8.0 mL of this solution to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity, discarding the first 10 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 243 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g/100 g, of C8H9NO2 in the portion of Acetaminophen for Effervescent Oral Solution taken: Result = (rU/rS) (CS/CU) F rU rS = peak response from the Sample solution = peak response from the Standard solution
10

= concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg CU Acetaminophen for Effervescent Oral Solution/mL) F = conversion factor (100) Acceptance criteria: 5.63 g6.88 g/100 g of C8H9NO2 CS
USP 32
Mode: LC Detector: UV 243 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in each Suppository taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature or in a cool place. USP REFERENCE STANDARDS 11 USP Acetaminophen RS rU rS CS
PERFORMANCE TESTS MINIMUM FILL 755 For solid packaged in multiple-unit containers: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905 For solid packaged in single-unit containers: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in air tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS
Acetaminophen Suppositories
(Comment on this Monograph)id=m197=Acetaminophen Suppositories=A-Monos.pdf) DEFINITION Acetaminophen Suppositories contain NLT 90.0% and NMT 110.0% of the labeled amount of acetaminophen (C8H9NO2). IDENTIFICATION A. The retention time of the chromatogram of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Transfer a portion of Suppositories, equivalent to 20 mg of acetaminophen, to a beaker, add 20 mL of methanol, and heat on a steam bath until melted. Remove the beaker from the steam bath, allow to cool with occasional stirring, and filter. Developing solvent system: Methylene chloride and methanol (4:1) Acceptance criteria: Meet the requirements ASSAY PROCEDURE Mobile phase: Methanol and water (1:3) Standard solution: 0.01 mg/mL of USP Acetaminophen RS in Mobile phase Sample stock solution: Tare a small dish and a glass rod, place in the dish NLT 5 Suppositories, heat gently on a steam bath until melted, then stir, cool while stirring, and weigh. Transfer a weighed portion of the mass, equivalent to 100 mg of acetaminophen, to a separator, add 30 mL of solvent hexane, and dissolve. Add 30 mL of water, shake gently, and allow the phases to separate. [NOTEIf an emulsion forms, allow sufficient time for it to separate.] Transfer the aqueous layer to a 200-mL volumetric flask, wash the solvent hexane in the separator with three 30-mL portions of water, adding the washings to the volumetric flask, and dilute with Mobile phase to volume. Sample solution: Transfer 5.0 mL of this Sample stock solution to a 250-mL volumetric flask, and dilute with Mobile phase to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity, discarding the first 10 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.)
Acetaminophen Oral Suspension

(Comment on this Monograph)id=m190=Acetaminophen Oral Suspension=A-Monos.pdf) DEFINITION Acetaminophen Oral Suspension is a suspension of Acetaminophen in a suitable aqueous vehicle. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C8H9NO2. IDENTIFICATION INFRARED ABSORPTION 197K Sample: Transfer a volume of Oral Suspension, equivalent to 240 mg of acetaminophen, to a separator, add 50 mL of ethyl acetate, and shake. Filter the ethyl acetate extract through a funnel containing glass wool and 10 g of anhydrous sodium sulfate. Collect the filtrate in a beaker, and evaporate on a steam bath to dryness. Dry the residue in a vacuum over silica gel. ASSAY PROCEDURE Mobile phase: Methanol and water (1:3) Standard solution: 0.01 mg/mL of USP Acetaminophen RS in Mobile phase Sample solution: Transfer a volume of Oral Suspension, previously well-shaken, equivalent to 100 mg of acetaminophen, to a 200-mL volumetric flask. Add 100 mL of Mobile phase, and shake by mechanical means for 10 min. Dilute with Mobile phase to volume. Transfer 5.0 mL of this solution to a 250-mL volumetric flask, and dilute with Mobile phase to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity, discarding the first 10 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 243 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in each mL of the Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for oral suspension packaged in single-unit containers DELIVERABLE VOLUME 698 Meets the requirements for oral suspension packaged in multiple-unit containers IMPURITIES Organic Impurities PROCEDURE: LIMIT OF 4-AMINOPHENOL Diluent: Methanol, formic acid, and water (75:2:425) Mobile phase: 0.01 M sodium butanesulfonate in Diluent Standard solution: 24 g/mL of USP 4-Aminophenol RS in Mobile phase Sample solution: Equivalent to 4.8 mg/mL of acetaminophen in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 272 nm Column: 4.6-mm 20-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L Analysis Samples: Standard solution and Sample solution Acceptance criteria: The peak area for 4-aminophenol from the Sample solution is not greater than the corresponding peak area from the Standard solution. SPECIFIC TESTS PH 791: 4.06.9 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP 4-Aminophenol RS rU rS CS

IDENTIFICATION A. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Equivalent to 1 mg/mL of acetaminophen from powdered Tablets in methanol; filtered Developing solvent system: Methylene chloride and methanol (4:1) Acceptance criteria: Meet the requirements ASSAY PROCEDURE Mobile phase: Methanol and water (1:3) Standard solution: 0.01 mg/mL of USP Acetaminophen RS in Mobile phase Sample solution: Weigh and finely powder NLT 20 Tablets. Dissolve a portion of the powder in Mobile phase to prepare a solution containing 0.01 mg/mL of acetaminophen. To facilitate dissolution, shake powder in Mobile phase by mechanical means for 10 min, and sonicate for 5 min before makeup with Mobile phase to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity, discarding the first 10 mL of the filtrate. Use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 243 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 243 nm Standard solution: USP Acetaminophen RS in Medium Sample solution: Sample per Dissolution 711. Analysis: Determine the amount of C8H9NO2 dissolved by using UV absorption on filtered portions of the Sample solution suitably diluted with Medium, if necessary, in comparison with a Standard solution having a known concentration of USP Acetaminophen RS. Acceptance criteria: NLT 80% (Q) of the labeled amount of C8H9NO2 For Tablets labeled as chewable Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL rU rS CS
Acetaminophen Tablets
(Comment on this Monograph)id=m200=Acetaminophen Tablets=A-Monos.pdf) DEFINITION Acetaminophen Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of acetaminophen (C8H9NO2).
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rS CS
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= peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1 Medium: Simulated gastric fluid TS (without enzyme); 900 mL Apparatus 2: 50 rpm Time: 15 min, 1 h, and 3 h Sample solution: Sample per Dissolution 711. Standard solution: USP Acetaminophen RS in Medium Spectrometric conditions Mode: UV Analytical wavelength: 280 nm Analysis: Determine the amount of C8H9NO2 dissolved from UV absorbances, using a filtered portion of the Sample solution in comparison with a Standard solution. Tolerances: The percentages of the labeled amount of C8H9NO2 dissolved at the times specified conform to Acceptance Table 2.
Time 15 min 1h 3h Amount Dissolved 45%65% 60%85% NLT 85%
Apparatus 2: 75 rpm Time: 45 min Detecor, Sample solution, Standard solution, and Analysis: Proceed as directed in Dissolution. Tolerances: NLT 75% (Q) of the labeled amount of C8H9NO2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label Tablets that must be chewed to indicate that they are to be chewed before swallowing. USP REFERENCE STANDARDS 11 USP Acetaminophen RS
Acetaminophen Extended-Release Tablets

(Comment on this Monograph)id=m205=Acetaminophen Extended-Release Tablets=A-Monos.pdf) DEFINITION Acetaminophen Extended-Release Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of acetaminophen (C8H9NO2). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Use a portion of powdered Tablets. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Phosphoric acid and water (1:9) Mobile phase: Methanol, Solution A, and water (300:1:700) Standard solution: Dissolve a quantity of USP Acetaminophen RS in methanol, and dilute with Mobile phase to 0.65 mg/mL. Sample stock solution: Transfer 10 Tablets into a 250-mL volumetric flask containing 50 mL of water and a magnetic stir bar. Stir at least 30 min or until the coating has dissolved. Add 150 mL of methanol, and stir for 45 min. Tablet cores should be disintegrated at least 15 min prior to ending the stirring. Remove the magnetic stir bar and rinse into the flask with methanol. Dilute with methanol to volume, and centrifuge, using the clear supernatant as the Sample stock solution. Sample solution: Dilute 5 mL of the Sample stock solution with Mobile phase to 200 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 295 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 3.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU = peak response from the Sample solution
For gelatin-coated Tablets Medium, Apparatus, Sample solution, Standard solution, Spectrometric conditions, and Analysis: Proceed as directed in Test 1. Times: 30 min, 90 min, and 4 h Tolerances: The percentage of the labeled amount of C8H9NO2 dissolved at the times specified conform to Acceptance Table 2.
Time 30 min 90 min 4h Amount Dissolved 40%60% 55%85% NLT 80%
Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Medium, Apparatus, Sample solution, Standard solution, Spectrometric conditions, and Analysis: Proceed as directed in Test 1. Times: 15 min, 1 h, and 3 h Tolerances: The percentages of the labeled amount of C8H9NO2 dissolved at the times specified conform to Acceptance Table 2.
Time 15 min 1h 3h Amount Dissolved 40%60% 55%75% NLT 80%
UNIFORMITY OF DOSAGE UNITS 905:
Meet the requirements
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where the Tablets are gelatin-coated, the label so states. When more than one Dissolution Test is given, the
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labeling states the Dissolution Test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Acetaminophen RS CS

= concentration of the corresponding USP Reference Standard in the Standard solution (mg/mL) = nominal concentration of the corresponding CU analyte in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amounts of C8H9NO2 and C9H8O4 PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Mobile phase, Solution A, and Chromatographic system: Prepare as directed in the Assay. Internal standard solution: 1 mg/mL of benzoic acid in methanol Sample stock solution: Sample per Dissolution 711. Sample solution: Combine 4.0 mL of the Sample stock solution and 1.0 mL of the Internal standard solution. Standard stock solution A: 70 g/mL USP Salicylic Acid RS in Solution A Standard solution A: Combine 4.0 mL of the Standard stock solution A and 1.0 mL of the Internal standard solution. Standard stock solution B: 360 g/mL each of USP Acetaminophen RS and USP Aspirin RS in Solution A Standard solution B: Combine 4.0 mL of the Standard stock solution B and 1.0 mL of the Internal standard solution. Analysis Samples: Sample solution, Standard solution A, and Standard solution B [NOTEThe relative retention times for acetaminophen, salicylic acid, aspirin, and benzoic acid are about 0.3, 0.4, 0.6, and 1.0, respectively.] Analyze using a 20-L injection size. Determine the amount of C8H9NO2 dissolved: Result = 90 (C/W) (RU/RS) = concentration of USP Acetaminophen RS in Standard solution B (g/mL) W = labeled amount of acetaminophen (mg) = relative peak response ratio from the Sample RU solution = relative peak response ratio from the Standard RS solution B Determine the amount of C9H8O4 dissolved: Result = {[90C1(RU1/RS1)] + [90C2(RU2/RS2)(1.3044)]}/W = concentration of USP Aspirin RS in Standard solution B (g/mL) RU1 = ratio of the relative peak response for the aspirin peak and the internal standard peak from the Sample solution RS1 = ratio of the relative peak response for the aspirin peak and the internal standard peak from Standard solution B C2 = concentration of USP Salicylic Acid RS in Standard solution A (g/mL) RU2 = ratio of the relative peak response for the salicylic acid peak and the internal standard peak from the Sample solution RS2 = ratio of the relative peak response for the salicylic acid peak and the internal standard peak from Standard solution A W = labeled amount of aspirin (mg) Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2 and C9H8O4 is dissolved. C1 C
Acetaminophen and Aspirin Tablets

(Comment on this Monograph)id=m230=Acetaminophen and Aspirin Tablets=A-Monos.pdf) DEFINITION Acetaminophen and Aspirin Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2) and aspirin (C9H8O4). IDENTIFICATION The retention times of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEInject the Standard solution and the Sample solution promptly after preparation.] Solution A: Chloroform, methanol, and glacial acetic acid (78:20:2) Mobile phase: Transfer 225 mg of tetramethylammonium hydroxide pentahydrate to a 1000-mL flask, and add 750 mL of water, 125 mL of methanol, 125 mL of acetonitrile, and 1.0 mL of glacial acetic acid. Stir for 3 min, and pass through a membrane filter having a 0.5-m or finer porosity. Internal standard solution: 20 mg/mL of benzoic acid in Solution A Standard solution: Transfer 325 mg of USP Acetaminophen RS and 325 mg of USP Aspirin RS to a 100-mL volumetric flask, add 10.0 mL of Internal standard solution, and dilute with Solution A to volume. Sample solution: Transfer an equivalent to 325 mg of acetaminophen, from finely powdered Tablets (NLT 20), to a 100-mL volumetric flask, add 10.0 mL of Internal standard solution and 50 mL of Solution A, and sonicate for 3 min. Dilute with Solution A to volume. Pass a portion of this solution through a filter having a 2.5-m or finer porosity and use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Sample: Standard solution [NOTEThe retention times for acetaminophen, salicylic acid (if present), aspirin, and benzoic acid are about 2, 3, 5, and 8 min, respectively.] Suitability requirements Relative standard deviation: NMT 3.0% for either analyte Analysis Samples: Standard solution and Sample solution Calculate, individually, the percentages of C8H9NO2 and C9H8O4 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 RU RS = ratio of the peak responses of the analyte and benzoic acid from the Sample solution = ratios of the peak responses of the analyte and benzoic acid from the Standard solution
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the labeled amount, in mg, of acetaminophen per Tablet; and J being the ratio of the labeled amount, in mg, of caffeine to the labeled amount, in mg, of acetaminophen per Tablet. Standard solution: Transfer 20.0 mL of the Standard stock solution and 3.0 mL of the Internal standard solution to a 50mL volumetric flask, and dilute with Solution A to volume. This solution contains 0.1 mg/mL of USP Acetaminophen RS, 0.1J mg/mL of USP Aspirin RS, and 0.1J mg/mL of USP Caffeine RS. Sample stock solution: Transfer an equivalent to 250 mg, from finely powdered Tablets (NLT 20), of acetaminophen to a 100-mL volumetric flask. Add 75 mL of Solution A and shake by mechanical means for 30 min. Dilute with Solution A to volume. Sample solution: Transfer 2.0 mL of the Sample stock solution to a 50-mL volumetric flask. Add 3.0 mL of Internal standard solution, and dilute with Solution A to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 10-cm; 5-m packing L1 Column temperature: 45 1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen, caffeine, aspirin, benzoic acid, and salicylic acid are about 0.3, 0.5, 0.8, 1.0, and 1.2, respectively.] Suitability requirements Tailing factor: NMT 1.2 for each analyte peak Resolution: NLT 1.4 between any of the analyte and internal standard peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate, individually, the percentages of C8H9NO2, C9H8O4, and C8H10N4O2 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = ratio of the peak responses of the analyte and internal standard peak from the Sample solution RS = ratio of the peak responses of the analyte and internal standard peak from the Standard solution CS = concentration of the corresponding USP Reference Standard in the Standard solution (mg/mL) = nominal concentration of the analyte in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 100 rpm Time: 60 min Mobile phase, Internal standard solution, Solution A, Standard stock solution, and Chromatographic system: Proceed as directed in the Assay. Sample stock solution: Sample per Dissolution 711. Sample solution: Transfer 20.0 mL of a filtered portion of the Sample stock solution to a 50-mL volumetric flask. Add 3.0 mL of Internal standard solution and 20 mL of Solution A. Mix, and allow to stand for 30 s. Dilute with Solution A to volume. Standard solution: Transfer 20.0 mL of Standard stock solution, 3.0 mL of Internal standard solution, and 20 mL of RU
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to acetaminophen and to aspirin IMPURITIES Organic Impurities PROCEDURE: LIMIT OF SALICYLIC ACID Solution A, Mobile phase, Internal standard solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay. Standard stock solution: 1.0 mg/mL of USP Salicylic Acid RS in Solution A Standard solutions: Transfer 1.0-mL, 5.0-mL, and 10.0-mL portions of Standard stock solution to separate 100-mL volumetric flasks, add 10.0 mL of Internal standard solution to each flask, and dilute with Solution A to volume. Analysis Samples: Sample solution and Standard solutions Plot the ratios of the peak responses for salicylic acid and benzoic acid for each of the Standard solutions versus concentrations, in mg/mL, of salicylic acid, and draw the straight line best fitting the three plotted points. From the graph so obtained, and from the ratio of the peak responses for salicylic acid and benzoic acid in the chromatogram of the Sample solution as obtained in the Assay, determine the concentration, in mg/mL, of salicylic acid (C7H6O3) in the Sample solution, and calculate the percentage of salicylic acid in relation to the concentration of aspirin in the Sample solution, as determined in the Assay. Acceptance criteria: NMT 3.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Aspirin RS USP Salicylic Acid RS
Acetaminophen, Aspirin, and Caffeine Tablets

(Comment on this Monograph)id=m240=Acetaminophen, Aspirin, and Caffeine Tablets=A-Monos.pdf) DEFINITION Acetaminophen, Aspirin, and Caffeine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), aspirin (C9H8O4), and caffeine (C8H10N4O2). IDENTIFICATION The retention times of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEInject the Standard solution and the Sample solution promptly after preparation.] Solution A: Methanol and glacial acetic acid (95:5) Mobile phase: Methanol, glacial acetic acid, and water (28:3:69) Internal standard solution: 6 mg/mL of benzoic acid in methanol Standard stock solution: Dissolve quantities of USP Acetaminophen RS, USP Aspirin RS, and USP Caffeine RS in Solution A to obtain a solution having known concentrations of 0.25 mg/mL of USP Acetaminophen RS, 0.25J mg/mL of USP Aspirin RS, and 0.25J mg/mL of USP Caffeine RS, J being the ratio of the labeled amount, in mg, of aspirin to
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water to a 50-mL volumetric flask. Mix, and allow to stand for about 30 s. Dilute with Solution A to volume. Use within 8 h. Analysis: Proceed as directed for Analysis in the Assay. Calculate, individually, the percentages of C8H9NO2, C9H8O4, and C8H10N4O2 dissolved: Result = (RU/RS) (CS/CU) 100 = ratio of the peak responses of the analyte and internal standard peak from the Sample solution RS = ratio of the peak responses of the analyte and internal standard peak from the Standard solution = concentration of the corresponding USP CS Reference Standard in the Standard solution (mg/mL) = nominal concentration of the analyte in the CU Sample solution (mg/mL) Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2, C9H8O4, and C8H10N4O2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to acetaminophen, aspirin, and caffeine RU IMPURITIES Organic Impurities PROCEDURE: LIMIT OF SALICYLIC ACID Mobile phase and Solution A: Prepare as directed in the Assay. Standard solution: 0.02 mg/mL of USP Salicylic Acid RS in Solution A Sample solution: Transfer an equivalent to 250 mg of aspirin, from finely powdered Tablets (NLT 20), to a 100mL volumetric flask. Add 75 mL of Solution A, and shake by mechanical means for 30 min. Dilute with Solution A to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 302 nm Column: 4.6-mm 10-cm; 5-m packing L1 Temperature: 45 1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.6 Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of salicylic acid in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) CU = nominal concentration of aspirin in the Sample solution (mg/mL) Acceptance criteria: NMT 3.0% rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Aspirin RS USP Caffeine RS

USP Salicylic Acid RS
Acetaminophen and Caffeine Tablets

(Comment on this Monograph)id=m246=Acetaminophen and Caffeine Tablets=A-Monos.pdf) DEFINITION Acetaminophen and Caffeine Tablets contain NLT 90.0% and NMT 110.0% of the labeled quantities of acetaminophen (C8H9NO2) and caffeine (C8H10N4O2). IDENTIFICATION The retention times of the Sample solution correspond to those of the Standard solution, relative to the internal standard, obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (28:3:69) Internal standard solution: 6 mg/mL of benzoic acid in methanol Solution A: Methanol and glacial acetic acid (95:5) Standard stock solution: 0.25 mg/mL of USP Acetaminophen RS and 0.25J mg/mL USP Caffeine RS in Solution A; J being the ratio of the labeled amount, in mg, of caffeine to the labeled amount, in mg, of acetaminophen per Tablet. Standard solution: Transfer 20.0 mL of Standard stock solution and 3.0 mL of Internal standard solution to a 50-mL volumetric flask, and dilute to volume with Solution A (0.1 mg/mL of USP Acetaminophen RS and 0.1J mg/mL of USP Caffeine RS). Sample stock solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 250 mg of acetaminophen, to a 100-mL volumetric flask. Add 75 mL of Solution A, and shake by mechanical means for 30 min. Dilute with Solution A to volume. Sample solution: Transfer 2.0 mL of the Sample stock solution and 3.0 mL of Internal standard solution to a 50-mL volumetric flask, and dilute with Solution A to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 10-cm; 5-m packing L1 Column temperature: 45 1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen, caffeine, and benzoic acid are about 0.3, 0.5, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.4 between any of the analyte and internal standard peaks Tailing factor: NMT 1.2 for each analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate, individually, the percentages of C8H9NO2 and C8H10N4O2 in the Tablets: Result = (RU/RS) (CS/CU) 100 RU = ratio of the peak responses of the analyte and internal standard peaks from the Sample solution
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RS Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine=A-Monos.pdf)
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= ratio of the peak responses of the analyte and internal standard peaks from the Standard solution = concentration of the corresponding USP CS Reference Standard in the Standard solution (mg/mL) = nominal concentration of the corresponding CU analyte in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of C8H9NO2 and C8H10N4O2 PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 100 rpm Time: 60 min Mobile phase, Internal standard solution, Solution A, Standard stock solution, and Chromatographic system: Proceed as directed in the Assay. Sample stock solution: Sample per Dissolution 711. Sample solution: Transfer an aliquot of a filtered portion of Sample stock solution to a 50-mL volumetric flask in order to obtain an expected concentration of 0.1 mg/mL of acetaminophen and 0.1J mg/mL of caffeine. Add 3.0 mL of Internal standard solution and 20 mL of Solution A, and allow to stand for 30 s. Dilute with Solution A to volume. [NOTEJ is defined for the Standard stock solution.] Standard solution: Transfer 20.0 mL of the Standard stock solution, 3.0 mL of Internal standard solution, and 20 mL of water to a 50-mL volumetric flask and allow to stand for 30 s. Dilute with Solution A to volume. Use within 8 h. Analysis: Proceed as directed for Analysis in the Assay, using the Sample solution and Standard solution prepared within Dissolution. Calculate the percentages of C8H9NO2 and C8H10N4O2 dissolved: Result = (RU/RS) (CS/CU) 100 = ratio of the peak responses of the analyte and internal standard peaks from the Sample solution = ratio of the peak responses of the analyte and RS internal standard peaks from the Standard solution = concentration of the corresponding USP CS Reference Standard in the Standard solution (mg/mL) CU = nominal concentration of the corresponding analyte in the Sample solution (mg/mL) Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2 and C8H10N4O2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Caffeine RS RU
DEFINITION Capsules Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO HBr H2O), and phenylpropanolamine hydrochloride (C9H13NO HCl). [NOTEThe heading of this monograph does not constitute the official title. It is not intended that the name described herein be recognized as the official title or the common or usual name. The name for each article encompassed by this monograph shall be composed of the names of the active ingredients contained therein, as well as the quantitative amount of each active ingredient, and a statement of the function (or purpose) of the ingredient in the article.] IDENTIFICATION A. If phenylpropanolamine hydrochloride is claimed in the labeling to be present, the chromatogram of the Sample solution, obtained as directed in the Assay for Phenylpropanolamine hydrochloride, exhibits a major peak for phenylpropanolamine, the retention time of which corresponds to that exhibited by the Standard solution. B. If acetaminophen is claimed in the labeling to be present, the chromatogram of the Sample solution, obtained as directed in the Assay for Acetaminophen, exhibits a major peak for acetaminophen, the retention time of which corresponds to that exhibited by the Standard solution. C. If chlorpheniramine maleate is claimed in the labeling to be present, the chromatogram of the Sample solution, obtained as directed in the Assay for Chlorpheniramine maleate, exhibits a major peak for chlorpheniramine, the retention time of which corresponds to that exhibited by the Standard solution. D. If dextromethorphan hydrobromide is claimed in the labeling to be present, the chromatogram of the Sample solution, obtained as directed in the Assay for Dextromethorphan hydrobromide, exhibits a major peak for dextromethorphan, the retention time of which corresponds to that exhibited by the Standard solution. ASSAY PHENYLPROPANOLAMINE HYDROCHLORIDE (if present) Mobile phase: Methanol and water (60:40) containing 0.34 g of monobasic potassium phosphate, 0.15 g of triethylamine hydrochloride, 0.25 g of sodium lauryl sulfate, and 0.1 mL of phosphoric acid in each 100 mL of solution Standard stock solution: 0.5 mg/mL USP Phenylpropanolamine Hydrochloride RS in Mobile phase Standard solution: 1 mL of Standard stock solution and 8 mL of Mobile phase. Dilute with water to 10 mL. Chlorpheniramine standard solution: Prepare as directed for Standard solution in the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Prepare as directed for Standard solution in the Assay for Dextromethorphan hydrobromide. System suitability solution 1 (for Capsules that contain chlorpheniramine salt): Standard solution and Chlorpheniramine standard solution (1:1) System suitability solution 2 (for Capsules that contain no chlorpheniramine salt): Standard solution and Dextromethorphan standard solution (1:1) Sample stock solution: Transfer NLT 10 Capsules to a 500mL volumetric flask. Add 100 mL of water and 10 mL of 5%
Capsules Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine
(Comment on this Monograph)id=m247=Capsules Containing at Least Three of the Following-Acetaminophen and Salts of
USP 32
phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, dilute with water to volume, and filter. Sample solution: Equivalent of 0.05 mg/mL of phenylpropanolamine hydrochloride from Sample stock solution diluted with water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution and System suitability solution 1 or System suitability solution 2 Suitability requirements Resolution: NLT 2.0 between phenylpropanolamine and chlorpheniramine or between phenylpropanolamine and dextromethorphan for the appropriate System suitability solution Tailing factor: NMT 2.0 for the phenylpropanolamine peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H13NO HCl: Result = (rU/rS) (CS/CU) 100 = phenylpropanolamine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Phenylpropanolamine CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of the CU phenylpropanolamine hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% ACETAMINOPHEN (if present) Mobile phase: Methanol, water, and glacial acetic acid (20:79:1) Standard solution: Transfer 25 mg of USP Acetaminophen RS to a 100-mL volumetric flask. Dissolve in 4 mL of methanol. Add 0.2 mL of phosphoric acid, and dilute with water to volume. Sample stock solution: Transfer NLT 10 Capsules to a 500mL volumetric flask. Add 100 mL of water and 10 mL of 5% phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, and dilute with water to volume. Sample solution: Equivalent of 0.25 mg/mL acetaminophen from Sample stock solution diluted with 0.1% phosphoric acid Chromatographic system (See Chromatography 621, System Suitability.) rU

Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2: Result = (rU/rS) (CS/CU) 100 = acetaminophen peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Acetaminophen RS in the CS Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Phenylpropanolamine hydrochloride. Standard stock solution: 0.8 mg/mL of USP Chlorpheniramine Maleate RS Standard solution: 8 g/mL of USP Chlorpheniramine Maleate RS from the Standard stock solution diluted with 0.1% phosphoric acid Sample stock solution: Transfer NLT 10 Capsules to a 500mL volumetric flask. Add 100 mL of water and 10 mL of 5% phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, dilute with water to volume, and filter. Sample solution: Equivalent of 8 g/mL of chlorpheniramine maleate from Sample stock solution diluted with 0.1% phosphoric solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4: rU Result = (rU/rs) (CS/CU) 100 rU rS CS CU = chlorpheniramine maleate peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlorpheniramine Maleate RS in the Standard solution (mg/mL) = nominal concentration of chlorpheniramine maleate in the Sample solution (mg/mL)
18
USP 32
Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Phenylpropanolamine hydrochloride. Standard stock solution: 0.6 mg/mL of USP Dextromethorphan Hydrobromide RS Standard solution: 0.06 mg/mL of USP Dextromethorphan Hydrobromide RS from Standard stock solution diluted with 0.1% phosphoric acid Sample stock solution: Transfer NLT 10 Capsules to a 500mL volumetric flask. Add 100 mL of water and 10 mL of 5% phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, dilute with water to volume, and filter. Sample solution: Equivalent of 0.06 mg/mL of dextromethorphan hydrobromide from Sample stock solution diluted with 0.1% phosphoric acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O: Result = (rU/rS) (CS/Cu) (Mr1/Mr2) 100 = dextromethorphan hydrobromide peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Dextromethorphan CS Hydrobromide RS in the Standard solution (mg/mL) CU = nominal concentration of dextromethorphan hydrobromide in the Sample solution (mg/mL) Mr1 = molecular weight of dextromethorphan hydrobromide monohydrate, 370.33 Mr2 = molecular weight of dextromethorphan hydrobromide anhydrous, 352.32 Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 1: 50 rpm Time: 45 min Sample solution: Mix 9.0 mL of a filtered portion of the solution under test with 1.0 mL of 1% phosphoric acid solution. Analysis: Determine the amounts of phenylpropanolamine hydrochloride, acetaminophen, chlorpheniramine maleate, and dextromethorphan hydrobromide dissolved, employing the procedures set forth in the Assay for Phenylpropanolamine hydrochloride, Assay for Acetaminophen, Assay for Chlorpheniramine maleate, and Assay for Dextromethorphan hydrobromide, respectively, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amounts of C9H13NO HCl, C8H9NO2, C16H19ClN2 C4H4O4, and C18H25NO HBr H2O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label for each article encompassed by this monograph bears a name composed of the active ingredients. The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS USP Dextromethorphan Hydrobromide RS USP Phenylpropanolamine Hydrochloride RS
Oral Solution Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine
(Comment on this Monograph)id=m248=Oral Solution Containing at Least Three of the Following-Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine=A-Monos.pdf) DEFINITION Oral Solution Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine contains NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8 H9NO2), chlorpheniramine maleate (C16H19 ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO HBr H2O), and phenylpropanolamine hydrochloride (C9H13NO HCl). [NOTEThe heading of this monograph does not constitute the official title. It is not intended that the name described herein be recognized as the official title or the common or usual name. The name for each article encompassed by this monograph shall be composed of the names of the active ingredients contained therein, as well as the quantitative amount of each active ingredient, and a statement of the function (or purpose) of the ingredient in the article.] IDENTIFICATION A. If phenylpropanolamine hydrochloride is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Phenylpropanolamine hydrochloride. B. If acetaminophen is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. C. If chlorpheniramine maleate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. D. If dextromethorphan hydrobromide is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan hydrobromide. ASSAY PHENYLPROPANOLAMINE HYDROCHLORIDE (if present) Mobile phase: 3.4 mg/mL of monobasic potassium phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric acid in methanol and water (60:40) Standard stock solution: 0.5 mg/mL of USP Phenylpropanolamine Hydrochloride RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:8:1) Chlorpheniramine standard solution: Use Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use Standard solution from the Assay for Dextromethorphan hydrobromide. System suitability solution A (for Oral Solution that contains either all the four ingredients or a combination of three containing chlorpheniramine salt): Standard solution and the Chlorpheniramine standard solution (1:1) System suitability solution B (for Oral Solution that contains no chlorpheniramine salt): Standard solution and Dextromethorphan standard solution (1:1)
USP 32
Sample solution: Equivalent to 0.05 mg/mL of phenylpropanolamine hydrochloride in Mobile phase and water (4:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: Standard solution, System suitability solution A (10 L), or System suitability solution B (10 L) Suitability requirements Resolution: NLT 2.0 between phenylpropanolamine and chlorpheniramine or between phenylpropanolamine and dextromethorphan, System suitability solution A or B Tailing factor: NMT 2.0 for the phenylpropanolamine peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H13NO HCl in the volume of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = phenylpropanolamine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Phenylpropanolamine CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of phenylpropanolamine CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% ACETAMINOPHEN (if present) Mobile phase: Methanol, glacial acetic acid, and water (20:1:79). Standard solution: Dissolve 16.5 mg of USP Acetaminophen RS in 2.5 mL of methanol, then dilute to 100 mL with water (0.165 mg/L). Sample solution: Equivalent to 0.165 mg/mL of acetaminophen in methanol and water (1:39) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the volume of Oral Solution taken: rU Result = (rU/rS) (CS/CU) 100 rU rS CS CU = acetaminophen peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of acetaminophen in the Sample solution (mg/mL)

Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Phenylpropanolamine hydrochloride. Standard stock solution: 0.08 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:8:1) Sample solution: Equivalent to 8 g/mL of acetaminophen maleate in Mobile phase and water (4:1) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in the volume of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = chlorpheniramine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Chlorpheniramine Maleate CS RS in the Standard solution (mg/mL) = nominal concentration of chlorpheniramine CU maleate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Phenylpropanolamine hydrochloride. Standard stock solution: 0.4 mg/mL of USP Dextromethorphan Hydrobromide RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:8:1) Sample solution: Equivalent to 0.04 mg/mL of dextromethorphan hydrobromide in Mobile phase and water (4:1) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in the volume of Oral Solution taken: rU Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = dextromethorphan peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Dextromethorphan CS Hydrobromide RS in the Standard solution (mg/mL) = nominal concentration of dextromethorphan CU hydrobromide in the Sample solution (mg/mL) = molecular weight of dextromethorphan Mr1 hydrobromide monohydrate, 370.33 = molecular weight of anhydrous Mr2 dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% rU OTHER COMPONENTS ALCOHOL DETERMINATION (if present), Method II 611: 90.0%110.0% of the labeled amount of C2 H5 OH SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Salmonella species and Escherichia coli. The total aerobic microbial count does not exceed 100 cfu/g, and the total combined molds and yeast count does not exceed 10 cfu/g.
20

2.67.5
USP 32
Standard stock solution: 2.5 mg/mL of USP Phenylpropanolamine Hydrochloride RS in water Standard solution: Standard stock solution, methanol, and 0.1% phosphoric acid (1:5:44) Chlorpheniramine standard solution: Use Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use Standard solution from the Assay for Dextromethorphan hydrobromide. System suitability solution A (for Tablets that contain either all the four ingredients or a combination of three containing chlorpheniramine salt): Standard solution and the Chlorpheniramine standard solution (1:1) System suitability solution B (for Tablets that contain no chlorpheniramine salt): Standard solution and Dextromethorphan standard solution (1:1) Sample solution: Transfer an equivalent to 2.5 mg of phenylpropanolamine hydrochloride, from finely powdered Tablets (NLT 20), to a 50-mL volumetric flask. Add 5 mL of methanol, and sonicate to disperse the powder. Dilute with 0.1% phosphoric acid to volume and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: Standard solution, System suitability solution A (10 L), or System suitability solution B (10 L) Suitability requirements Resolution: NLT 2.0 between phenylpropanolamine and chlorpheniramine or between phenylpropanolamine and dextromethorphan, System suitability solution A or B Tailing factor: NMT 2.0 for the phenylpropanolamine peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H13NO HCl in portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = phenylpropanolamine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Phenylpropanolamine CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of phenylpropanolamine CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% ACETAMINOPHEN (if present) Mobile phase: Methanol, glacial acetic acid, and water (20:1:79). Standard solution: Dissolve 50 mg of USP Acetaminophen RS in 4 mL of methanol, then dilute with 0.1% phosphoric acid to 100 mL. Sample solution: Transfer an equivalent to 100 mg of acetaminophen, from finely powdered Tablets (NLT 20), to a 50-mL volumetric flask. Add 7.5 mL of methanol, and sonicate to disperse the powder. Add 0.5 mL of phosphoric acid, dilute with water to volume, and filter. Transfer 25.0 mL of the filtered solution to a 100-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) rU
PH 791:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label for each article encompassed by this monograph bears a name composed of the active ingredients. The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS USP Dextromethorphan Hydrobromide RS USP Phenylpropanolamine Hydrochloride RS
Tablets Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine
(Comment on this Monograph)id=m255=Tablets Containing at Least Three of the Following-Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine=A-Monos.pdf) DEFINITION Tablets Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Phenylpropanolamine contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO HBr H2O), and phenylpropanolamine hydrochloride (C9H13NO HCl). [NOTEThe heading of this monograph does not constitute the official title. It is not intended that the name described herein be recognized as the official title or the common or usual name. The name for each article encompassed by this monograph shall be composed of the names of the active ingredients contained therein as well as the quantitative amount of each active ingredient, and a statement of the function (or purpose) of the ingredient in the article.] IDENTIFICATION A. If phenylpropanolamine hydrochloride is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Phenylpropanolamine hydrochloride. B. If acetaminophen is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. C. If chlorpheniramine maleate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. D. If dextromethorphan hydrobromide is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan hydrobromide. ASSAY PHENYLPROPANOLAMINE HYDROCHLORIDE Mobile phase: 3.4 mg/mL of monobasic potassium phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric acid in methanol and water (60:40)
USP 32
Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = acetaminophen peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Acetaminophen RS in the CS Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 0.8 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: 8 g/mL of USP Chlorpheniramine Maleate RS from Standard stock solution in 0.1% phosphoric acid Sample solution: Transfer an equivalent to 0.4 mg of chlorpheniramine maleate, from finely powdered Tablets (NLT 20), to a 50-mL volumetric flask. Add 5 mL of methanol, and sonicate to disperse the powder. Add 0.2 mL of phosphoric acid, dilute with water to volume, and filter. Injection size: 20 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in the portion of Tablets taken: rU Result = (rU/rS) (CS/CU) 100 = chlorpheniramine peak response from the Sample solution rS = peak response from the Standard solution = concentration of USP Chlorpheniramine Maleate CS RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Phenylpropanolamine hydrochloride. Standard stock solution: 0.6 mg/mL of USP Dextromethorphan Hydrobromide RS in water Standard solution: 60 g/mL of USP Dextromethorphan Hydrobromide RS from Standard stock solution in 0.1% phosphoric acid Sample solution: Transfer an equivalent to 3 mg of dextromethorphan hydrobromide, from finely powdered Tablets (NLT 20), to a 50-mL volumetric flask. Add 5 mL of methanol, and sonicate to disperse the powder. Add 0.2 mL of phosphoric acid, dilute with water to volume, and filter. rU

Injection size: 20 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = dextromethorphan peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Dextromethorphan CS Hydrobromide RS in the Standard solution (mg/mL) = nominal concentration of dextromethorphan CU hydrobromide in the Sample solution (mg/mg) Mr1 = molecular weight of dextromethorphan hydrobromide monohydrate, 370.33 = molecular weight of anhydrous Mr2 dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: 0.1 M hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 45 min Sample solution: Mix 9.0 mL of a filtered portion of the solution under test with 1.0 mL of 1% phosphoric acid solution. Analysis: Determine the amounts of phenylpropanolamine hydrochloride, acetaminophen, chlorpheniramine maleate, and dextromethorphan hydrobromide dissolved, employing the Analyses set forth in the Assays for Phenylpropanolamine hydrochloride, Acetaminophen, Chlorpheniramine maleate, and Dextromethorphan hydrobromide, respectively, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amount of C9H13NO HCl, C8H9NO2, C16H19ClN2 C4H4O4, and C18H25NO HBr H2O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label for each article encompassed by this monograph bears a name composed of the active ingredients. The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS USP Dextromethorphan Hydrobromide RS USP Phenylpropanolamine Hydrochloride RS
Capsules Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine
(Comment on this Monograph)id=m249=Capsules Containing at Least Three of the Following-Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine=A-Monos.pdf) DEFINITION Capsules Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine,
22
USP 32
Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: Standard solution and System suitability solution A or System suitability solution B Suitability requirements Resolution: NLT 2.0 between pseudoephedrine and chlorpheniramine or between pseudoephedrine and dextromethorphan, System suitability solution A or B Tailing factor: NMT 2.5 for pseudoephedrine peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H15NO HCl in the Capsules taken: Result = (rU/rS) (CS/CU) 100 = pseudoephedrine peak response from the Sample solution rS = peak response from the Standard solution = concentration of USP Pseudoephedrine CS Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is the salt form used, if present in the formulation) Mobile phase, System suitability solutions A and B, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Chlorpheniramine standard solution: Use Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use Standard solution from the Assay for Dextromethorphan hydrobromide. Standard stock solution: 3.0 mg/mL of USP Pseudoephedrine Sulfate RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:4:5) Sample solution: Proceed as directed for the Sample solution in the Assay for Pseudoephedrine hydrochloride to obtain a solution having a concentration of 0.24 mg/mL of pseudoephedrine sulfate. Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C10H15NO)2 H2SO4 in the Capsules taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = pseudoephedrine peak response from the Sample solution = peak response from the Standard solution = concentration of USP Pseudoephedrine Sulfate RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL) rU
Dextromethorphan, and Pseudoephedrine contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO HBr H2O), and pseudoephedrine hydrochloride (C10H15NO HCl) or pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. [NOTEThe heading of this monograph does not constitute the official title. It is not intended that the name described herein be recognized as the official title or the common or usual name. The name for each article encompassed by this monograph shall be composed of the names of the active ingredients contained therein, as well as the quantitative amount of each active ingredient, and a statement of the function (or purpose) of the ingredient in the article.] IDENTIFICATION A. If pseudoephedrine hydrochloride or pseudoephedrine sulfate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Pseudoephedrine sulfate. B. If acetaminophen is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. C. If chlorpheniramine maleate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. D. If dextromethorphan hydrobromide is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan hydrobromide. ASSAY PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine hydrochloride is the salt form used, if present in the formulation) Mobile phase: 3.4 mg/mL of monobasic potassium phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric acid in methanol and water (60:40). Standard stock solution: 1.5 mg/mL of USP Pseudoephedrine Hydrochloride RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:8:1) Chlorpheniramine standard solution: Use Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use Standard solution from the Assay for Dextromethorphan hydrobromide. System suitability solution A (for Oral Solution that contains either all the four ingredients or a combination of three containing chlorpheniramine salt): Standard solution and Chlorpheniramine standard solution (1:1) System suitability solution B (for Oral Solution that contains no chlorpheniramine salt): Standard solution and Dextromethorphan standard solution (1:1) Sample stock solution: Transfer NLT 10 Capsules to a 500mL volumetric flask. Add 100 mL of water and 10 mL of 5% phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, dilute with water to volume, and filter. Sample solution: 0.12 mg/mL of pseudoephedrine hydrochloride, from Sample stock solution in water Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Acceptance criteria: 90.0%110.0% ACETAMINOPHEN (if present) Mobile phase: Methanol, glacial acetic acid, and water (20:1:79) Standard solution: Dissolve 25 mg of USP Acetaminophen RS in 4 ml of methanol, then add 0.2 mL of phosphoric acid and dilute to 100 mL with water (0.25 mg/mL). Sample solution: Transfer NLT 10 Capsules to a 500-mL volumetric flask. Add 100 mL of water and 10 mL of 5% phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, and dilute with water to volume. Quantitatively dilute a portion of this solution, if necessary, with 0.1% phosphoric acid to obtain a solution having a concentration of 0.25 mg/mL of acetaminophen. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the Capsules taken: Result = (rU/rS) (CS/CU) 100 = acetaminophen peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Acetaminophen RS in the CS Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 1 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:80:19) Sample solution: Transfer NLT 10 Capsules to a 500-mL volumetric flask. Add 100 mL of water and 10 mL of 5% phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, dilute with water to volume, and filter. Quantitatively dilute a portion of this solution, if necessary, with 0.1% phosphoric acid to obtain a solution having a concentration of 8 g/mL of chlorpheniramine maleate. Injection size: 10 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in the Capsules taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = chlorpheniramine peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlorpheniramine Maleate RS in the Standard solution (mg/mL) rU CU

= nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 1.5 mg/mL of USP Dextromethorphan Hydrobromide RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:16:3) Sample solution: Transfer NLT 10 Capsules to a 500-mL volumetric flask. Add 100 mL of water and 10 mL of 5% phosphoric acid, and gently heat until the Capsules are fully dispersed. Cool the solution to room temperature, dilute with water to volume, and filter. Quantitatively dilute a portion of this solution, if necessary, with 0.1% phosphoric acid to obtain a solution having a concentration of 0.04 mg/mL of dextromethorphan hydrobromide. Injection size: 10 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in the Capsules taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = dextromethorphan peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Dextromethorphan CS Hydrobromide RS in the Standard solution (mg/mL) CU = nominal concentration of dextromethorphan hydrobromide in the Sample solution (mg/mL) Mr1 = molecular weight of dextromethorphan hydrobromide monohydrate, 370.33 = molecular weight of anhydrous Mr2 dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Sample solution: Mix 9.0 mL of a filtered portion of the solution under test with 1.0 mL of 1% phosphoric acid solution. Analysis: Determine the amounts of pseudoephedrine hydrochloride or pseudoephedrine sulfate (as appropriate), acetaminophen, chlorpheniramine maleate, and dextromethorphan hydrobromide dissolved, employing the Analyses set forth in the Assays for Pseudoephedrine hydrochloride or Pseudoephedrine sulfate, Acetaminophen, Chlorpheniramine maleate, and Dextromethorphan hydrobromide, respectively, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amounts of C10H15NO HCl or (C10H15 NO)2 H2SO4, C8H9NO2, C16H19ClN2 C4H4O4, and C18H25NO HBr H2O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: The label for each article encompassed by this monograph bears a name composed of the active ingredients contained in the article. The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article.
24
USP 32
Standard solution: 0.24 mg/mL by diluting Standard stock solution with 0.1% phosphoric acid Chlorpheniramine standard solution: Prepare as directed for Standard solution in the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Prepare as directed for Standard solution in the Assay for Dextromethorphan hydrobromide. System suitability solution A (for Oral Powder that contains either all of the four ingredients or a combination of three that includes chlorpheniramine salt): Standard solution and the Chlorpheniramine standard solution (1:1) System suitability solution B (for Oral Powder that contains no chlorpheniramine salt): Standard solution and the Dextromethorphan standard solution (1:1) Sample stock solution: Transfer the contents of 10 unitdose containers of the Oral Powder to a 2000-mL volumetric flask. Add 1000 mL of water and 2.0 mL of phosphoric acid. Gently heat to 60 until the powder is fully dispersed. Cool the flask to room temperature, add 40 mL of methanol, and dilute with water to volume. Sample solution: Equivalent of 0.24 mg/mL of pseudoephedrine hydrochloride from Sample stock solution in 0.1% phosphoric acid Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection volume: 10 L System suitability Samples: Standard solution and System suitability solution A or System suitability solution B Suitability requirements Resolution: NLT 2.0 between pseudoephedrine and chlorpheniramine or between pseudoephedrine and dextromethorphan, System suitability solution A or System suitability solution B Tailing factor: NMT 2.5 for the pseudoephedrine peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H15NO HCl in each unitdose container of Oral Powder taken: Result = (rU/rS) (CS/CU) 100 = peak response for pseudoephedrine hydrochlorde from the Sample solution rS = peak response from the Standard solution CS = concentration of USP Pseudoephedrine Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of pseudoephedrine hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is the salt form used, if present in the formulation) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Chlorpheniramine standard solution: Prepare as directed for Standard solution in the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Prepare as directed for Standard solution in the Assay for Dextromethorphan hydrobromide. rU
USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS USP Dextromethorphan Hydrobromide RS USP Pseudoephedrine Hydrochloride RS USP Pseudoephedrine Sulfate RS
Oral Powder Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine
(Comment on this Monograph)id=m251=Oral Powder Containing at Least Three of the Following-Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine=A-Monos.pdf) DEFINITION Oral Powder Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine contains NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO HBr H2O), and pseudoephedrine hydrochloride (C10H15NO HCl) or pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. [NOTEThe heading of this monograph does not constitute the official title. It is not intended that the name described herein be recognized as the official title or the common or usual name. The name for each article encompassed by this monograph shall be composed of the names of the active ingredients contained therein, as well as the quantitative amount of each active ingredient, and a statement of the function (or purpose) of the ingredient in the article.] IDENTIFICATION A. If pseudoephedrine hydrochloride or pseudoephedrine sulfate is claimed in the labeling to be present, the retention time of the major peak for pseudoephedrine of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Pseudoephedrine hydrochloride or the Assay for Pseudoephedrine sulfate. B. If acetaminophen is claimed in the labeling to be present, the retention time of the major peak for acetaminophen of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. C. If chlorpheniramine maleate is claimed in the labeling to be present, the retention time of the major peak for chlorpheniramine of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. D. If dextromethorphan hydrobromide is claimed in the labeling to be present, the retention time of the major peak for dextromethorphan of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan hydrobromide. ASSAY PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine hydrochloride is the salt form used, if present in the formulation) Mobile phase: Mixture of methanol and water (60:40) containing 0.34 g of monobasic potassium phosphate, 0.3 g of triethylamine hydrochloride, 0.15 g of sodium lauryl sulfate, and 0.1 mL of phosphoric acid in each 100 mL of solution Standard stock solution: 3.0 mg/mL of USP Pseudoephedrine Hydrochloride RS
USP 32
Standard stock solution: 6.0 mg/mL of USP Pseudoephedrine Sulfate RS Standard solution: 0.48 mg/mL by diluting Standard stock solution with 0.1% phosphoric acid Sample solution: Proceed as directed for the Sample solution in the Assay for Pseudoephedrine hydrochloride to obtain a solution having a nominal concentration of 0.48 mg/mL of pseudoephedrine sulfate. Analysis: Proceed as directed for Analysis in the Assay for Pseudoephedrine hydrochloride. Calculate the percentage of (C10H15NO)2 H2SO4 in each unit-dose container of Oral Powder taken: Result = (rU/rS) (CS/CU) 100 = peak response for pseudoephedrine hydrochloride from the Sample solution = peak response from the Standard solution rS = concentration of USP Pseudoephedrine Sulfate RS CS in the Standard solution (mg/mL) = nominal concentration of pseudoephedrine CU sulfate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% ACETAMINOPHEN (if present) Mobile phase, Standard solution, and Chromatographic system: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Sample stock solution: Transfer the contents of 10 unitdose containers of the Oral Powder to a 2000-mL volumetric flask. Add 1000 mL of water and 2 mL of phosphoric acid. Gently heat to 60 until the powder is fully dispersed. Cool the flask to room temperature, add 40 mL of methanol, and dilute with water to volume. Sample solution: Nominally 0.50 mg/mL of acetaminophen from Sample stock solution and 0.1% phosphoric acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in each unit-dose container of Oral Powder taken: rU Result = (rU/rS) (CS/CU) x100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE (if present) Mobile phase and Chromatographic system: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 0.8 mg/mL of USP Chlorpheniramine Maleate RS Standard solution: 8 g/mL of chlorpheniramine maleate from Standard stock solution and 0.1% phosphoric acid Sample stock solution: Transfer the contents of 10 unitdose containers of Oral Powder to a 2000-mL volumetric flask. Add 1000 mL of water and 2 mL of phosphoric acid. Gently heat to 60 until the powder is fully dispersed. Cool the flask to room temperature, add 40 mL of methanol, dilute with water to volume, and mix. Sample solution: Nominally 8 g/mL solution of chlorpheniramine maleate from Sample stock solution and 0.1% phosphoric acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in each unit-dose container of Oral Powder taken: rU rS CS Result = (rU/rS) (CS/CU) 100 rU rS CS

= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlorpheniramine Maleate RS in the Standard solution (g/mL) = nominal concentration of chlorpheniramine CU maleate in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine Hydrochloride. Standard stock solution: 0.8 mg/mL of USP Dextromethorphan Hydrobromide RS Standard solution: 0.08 mg/mL solution from Standard stock solution and 0.1% phosphoric acid Sample stock solution: Transfer the contents of 10 unitdose containers of Oral Powder to a 2000-mL volumetric flask. Add 1000 mL of water and 2 mL of phosphoric acid. Gently heat to 60 until the powder is fully dispersed. Cool the flask to room temperature, add 40 mL of methanol, and dilute with water to volume. Sample solution: Nominally 0.08 mg/mL of dextromethorphan hydrobromide from Sample stock solution and 0.1% phosphoric acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in each unit-dose container of Oral Powder taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response for dextromethorphan hydrobromide from the Sample solution = peak response from the Standard solution rS = concentration of USP Dextromethorphan CS Hydrobromide RS in the Standard solution (mg/mL) = nominal concentration of dextromethorphan CU hydrobromide in the Sample solution (mg/mL) = molecular weight of dextromethorphan Mr1 hydrobromide monohydrate, 370.33 Mr2 = molecular weight of anhydrous dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 For oral powder packaged in single-unit containers: Meets the requirements MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: The label for each article encompassed by this monograph bears a name composed of the active ingredients. The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS USP Dextromethorphan Hydrobromide RS USP Pseudoephedrine Hydrochloride RS USP Pseudoephedrine Sulfate RS rU
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USP 32
Sample solution: Equivalent to 0.15 mg/mL of pseudoephedrine hydrochloride from Oral solution in Mobile phase and water (4:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: Standard solution and System suitability solution A or System suitability solution B Suitability requirements Resolution: NLT 2.0 between pseudoephedrine and chlorpheniramine or between pseudoephedrine and dextromethorphan, System suitability solution A or B Tailing factor: NMT 2.5 for pseudoephedrine peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H15NO HCl in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = pseudoephedrine peak response from the Standard solution CS = concentration of USP Pseudoephedrine Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of pseudoephedrine hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is the salt form used, if present in the formulation) Mobile phase, System suitability solutions A and B, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Chlorpheniramine standard solution: Use Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use Standard solution from the Assay for Dextromethorphan hydrobromide. Standard stock solution: 3.0 mg/mL of USP Pseudoephedrine Sulfate RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:4:5) Sample solution: Equivalent to 0.3 mg/mL of pseudoephedrine sulfate in Mobile phase and water (4:1) Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C10H15NO)2 H2SO4 in each mL of Oral Solution taken: rU rS Result = (rU/rS) (CS/CU) 100 = pseudoephedrine peak response from the Sample solution rS = peak response from the Standard solution CS = concentration of USP Pseudoephedrine Sulfate RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% ACETAMINOPHEN (if present) Mobile phase: Methanol, glacial acetic acid, and water (20:1:79) rU
Oral Solution Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine
(Comment on this Monograph)id=m254=Oral Solution Containing at Least Three of the Following-Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine=A-Monos.pdf) DEFINITION Oral Solution Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine contains NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO HBr H2O), and pseudoephedrine hydrochloride (C10H15NO HCl) or pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. [NOTEThe heading of this monograph does not constitute the official title. It is not intended that the name described herein be recognized as the official title or the common or usual name. The name for each article encompassed by this monograph shall be composed of the names of the active ingredients contained therein, as well as the quantitative amount of each active ingredient, and a statement of the function (or purpose) of the ingredient in the article.] IDENTIFICATION A. If pseudoephedrine hydrochloride or pseudoephedrine sulfate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Pseudoephedrine sulfate. B. If acetaminophen is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. C. If chlorpheniramine maleate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. D. If dextromethorphan hydrobromide is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan hydrobromide. ASSAY PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine hydrochloride is the salt form used, if present in the formulation) Mobile phase: 3.4 mg/mL of monobasic potassium phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric acid in methanol and water (60:40) Standard stock solution: 1.5 mg/mL of USP Pseudoephedrine Hydrochloride RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:8:1) Chlorpheniramine standard solution: Use Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use Standard solution from the Assay for Dextromethorphan hydrobromide. System suitability solution A (for Oral Solution that contains either all the four ingredients or a combination of three containing chlorpheniramine salt): Standard solution and Chlorpheniramine standard solution (1:1) System suitability solution B (for Oral Solution that contains no chlorpheniramine salt): Standard solution and Dextromethorphan standard solution (1:1)
USP 32
Standard solution: 0.165 mg/mL of USP Acetaminophen RS, in methanol and water (1:39) [NOTEFirst dissolve in methanol, and mix until dissolved and then dilute with water to volume.] Sample solution: Equivalent to 0.165 mg/mL of acetaminophen, from Oral Solution in methanol and water (1:39) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in each mL of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = acetaminophen peak response from the Sample solution rS = peak response from the Standard solution = concentration of USP Acetaminophen RS in the CS Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 1 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:80:19) Sample solution: Equivalent to 0.01 mg/mL of chlorpheniramine maleate from Oral Solution in Mobile phase and water (4:1) Injection size: 10 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = chlorpheniramine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Chlorpheniramine Maleate CS RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 1.5 mg/mL of USP Dextromethorphan Hydrobromide RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:16:3) rU rU

Sample solution: Equivalent to 0.075 mg/mL of dextromethorphan hydrobromide, from Oral Solution in Mobile phase and water (4:1) Injection size: 10 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in each mL of Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = dextromethorphan peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Dextromethorphan CS Hydrobromide RS in the Standard solution (mg/mL) = nominal concentration of dextromethorphan CU hydrobromide in the Sample solution (mg/mL) = molecular weight of dextromethorphan Mr1 hydrobromide monohydrate, 370.33 = molecular weight of anhydrous Mr2 dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% rU OTHER COMPONENTS ALCOHOL DETERMINATION (if present), Method II 611: 90.0%110.0% of the labeled amount of C2H5OH PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 For Oral Solution packaged in single-unit containers: Meets the requirements DELIVERABLE VOLUME 698 For Oral Solution packaged in multiple-unit containers: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: The total bacterial count does not exceed 100 cfu/g, the total combined molds and yeasts count does not exceed 10 cfu/g, and it meets the requirements of the tests for absence of Salmonella species and Escherichia coli. PH 791: 3.77.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: The label for each article encompassed by this monograph bears a name composed of the active ingredients. The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS USP Dextromethorphan Hydrobromide RS USP Pseudoephedrine Hydrochloride RS USP Pseudoephedrine Sulfate RS
Tablets Containing at Least Three of the FollowingAcetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine
(Comment on this Monograph)id=m256=Tablets Containing at Least Three of the Following-Acetaminophen and Salts of
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USP 32
Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 10 l System suitability Samples: Standard solution and System suitability solution A or System suitability solution B Suitability requirements Resolution: NLT 2.0 between pseudoephedrine and chlorpheniramine or between pseudoephedrine and dextromethorphan Tailing factor: NMT 2.5 for pseudoephedrine peak, Standard solution Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H15NO HCl in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = pseudoephedrine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Pseudoephedrine CS Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE SULFATE (where pseudoephedrine sulfate is the salt form used, if present in the formulation) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Chlorpheniramine standard solution: Use the Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use the Standard solution from the Assay for Dextromethorphan hydrobromide. Standard stock solution: 3.0 mg/mL of USP Pseudoephedrine Sulfate RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:4:5) Sample solution: Equivalent to 0.24 mg/mL of pseudoephedrine sulfate, from powdered Tablets, in methanol and 0.1% phosphoric acid (1:9) [NOTEFinely powder NLT 20 Tablets. First add methanol, and sonicate to disperse the powder, and then make up to volume with 0.1% phosphoric acid.] Analysis: Calculate the percentage of (C10H15NO)2 H2SO4 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = pseudoephedrine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Pseudoephedrine Sulfate RS CS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% ACETAMINOPHEN (if present) Mobile phase: Methanol, glacial acetic acid, and water (20:1:79) rU rU
Chlorpheniramine, Dextromethorphan, and Pseudoephedrine=A-Monos.pdf) DEFINITION Tablets Containing at Least Three of the Following Acetaminophen and Salts of Chlorpheniramine, Dextromethorphan, and Pseudoephedrine contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), dextromethorphan hydrobromide (C18H25NO HBr H2O), and pseudoephedrine hydrochloride (C10H15NO HCl) or pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. [NOTEThe heading of this monograph does not constitute the official title. It is not intended that the name described herein be recognized as the official title or the common or usual name. The name for each article encompassed by this monograph shall be composed of the names of the active ingredients contained therein, as well as the quantitative amount of each active ingredient, and a statement of the function (or purpose) of the ingredient in the article.] IDENTIFICATION A. If pseudoephedrine hydrochloride or pseudoephedrine sulfate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Pseudoephedrine sulfate. B. If acetaminophen is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. C. If chlorpheniramine maleate is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. D. If dextromethorphan hydrobromide is claimed in the labeling to be present, the retention time of the peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan hydrobromide. ASSAY PSEUDOEPHEDRINE HYDROCHLORIDE (where pseudoephedrine hydrochloride is the salt form used, if present in the formulation) Mobile phase: 3.4 mg/mL of monobasic potassium phosphate, 1.5 mg/mL of triethylamine hydrochloride, 2.5 mg/mL of sodium lauryl sulfate, and 1L/mL of phosphoric acid in methanol and water (60:40) Standard stock solution: 1.5 mg/mL of USP Pseudoephedrine Hydrochloride RS in water Standard solution: Mobile phase, Standard stock solution, and water (8:1:1) Chlorpheniramine standard solution: Use the Standard solution from the Assay for Chlorpheniramine maleate. Dextromethorphan standard solution: Use the Standard solution from the Assay for Dextromethorphan hydrobromide. System suitability solution A (for Oral Solution that contains either all the four ingredients or a combination of three containing chlorpheniramine salt): Standard solution and Chlorpheniramine standard solution (1:1) System suitability solution B (for Oral Solution that contains no chlorpheniramine salt): Standard solution and Dextromethorphan standard solution (1:1) Sample solution: Equivalent to 0.12 mg/mL of pseudoephedrine hydrochloride, from powdered Tablets, in methanol and 0.1% phosphoric acid (1:9) [NOTEFinely powder NLT 20 Tablets. First add methanol, and sonicate to disperse the powder, and then make up to volume with 0.1% phosphoric acid.] Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Standard solution: 0.165 mg/mL of USP Acetaminophen RS in methanol and water (1:39) [NOTEFirst dissolve in methanol, and mix until solution is complete, and then dilute with water to volume.] Sample solution: Equivalent to 0.165 mg/mL of acetaminophen, from Oral Solution in methanol and water (1:39) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = acetaminophen peak response from the Sample solution rS = peak response from the Standard solution = concentration of USP Acetaminophen RS in the CS Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE (if present) Mobile phase, System suitability solutions, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 1 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:80:19) Sample solution: Equivalent to 0.01 mg/mL of chlorpheniramine maleate from Oral Solution in Mobile phase and water (4:1) Injection size: 10 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in each mL of the Oral Solution: Result = (rU/rS) (CS/CU) 100 = chlorpheniramine peak response from the Sample solution = peak response from the Standard solution rS = concentration of USP Chlorpheniramine Maleate CS RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE (if present) Mobile phase, Chromatographic system, and System suitability: Proceed as directed in the Assay for Pseudoephedrine hydrochloride. Standard stock solution: 1.5 mg/mL of USP Dextromethorphan Hydrobromide RS in water Standard solution: Standard stock solution, Mobile phase, and water (1:16:3) rU rU

Sample solution: Equivalent to 0.075 mg/mL of dextromethorphan hydrobromide, from Oral solution in Mobile phase and water (4:1) Injection size: 10 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = dextromethorphan peak response from the Sample solution = dextromethorphan peak response from the rS Standard solution = concentration of USP Dextromethorphan CS Hydrobromide RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of dextromethorphan Mr1 hydrobromide monohydrate, 370.33 = molecular weight of anhydrous Mr2 dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION 711 Test 1 Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions ); 900 mL Apparatus 2: 50 rpm Time: 45 min Sample solution: Mix 9.0 mL of a filtered portion of the solution under test with 1.0 mL of 1% phosphoric acid solution. Analysis: Determine the amounts of pseudoephedrine hydrochloride or pseudoephedrine sulfate (as appropriate), acetaminophen, chlorpheniramine maleate, and dextromethorphan hydrobromide dissolved, using the Analyses set forth in the Assay for Pseudoephedrine hydrochloride or Assay for Pseudoephedrine sulfate, Assay for Acetaminophen, Assay for Chlorpheniramine maleate, and Assay for Dextromethorphan hydrobromide, respectively, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amounts of pseudoephedrine hydrochloride or pseudoephedrine sulfate, acetaminophen, chlorpheniramine maleate, and dextromethorphan hydrobromide Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Medium: Water; 900 mL Apparatus, Time, Sample solution, Analysis, and Tolerances: Proceed as directed for Test 1. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: The label for each article encompassed by this monograph bears a name composed of the active ingredients. The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article. When more than one Dissolution Test is given, the labeling states the Dissolution Test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS
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USP Dextromethorphan Hydrobromide RS USP Pseudoephedrine Hydrochloride RS USP Pseudoephedrine Sulfate RS CS
USP 32
= concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% CHLORPHENIRAMINE MALEATE Mobile phase: Methanol and water (3:2) containing 0.34 g of monobasic potassium phosphate, 0.3 g of triethylamine hydrochloride, 0.15 g of sodium lauryl sulfate, and 0.1 mL of phosphoric acid in each 100 mL of solution Pseudoephedrine hydrochloride standard stock solution: 3.0 mg/mL of USP Pseudoephedrine Hydrochloride RS Pseudoephedrine hydrochloride standard solution: Transfer 1.0 mL of Pseudoephedrine hydrochloride standard stock solution to a 25-mL volumetric flask. Add 2.5 mL of methanol, and dilute with 0.1% phosphoric acid to volume. Standard stock solution: 0.8 mg/mL of USP Chlorpheniramine Maleate RS Standard solution: 8 g/mL of USP Chlorpheniramine Maleate RS in 0.1% phosphoric acid, from Standard stock solution System suitability solution: Pseudoephedrine hydrochloride stock solution and Standard solution (1:1) Sample solution: Transfer an equivalent to 2 mg of chlorpheniramine maleate, from powdered Tablets (NLT 20), to a 250-mL volumetric flask. Add 25 mL of methanol, and sonicate to disperse the powder. Add 1 mL of phosphoric acid, dilute with water to volume, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 2.0 between pseudoephedrine and chlorpheniramine, System suitability solution Tailing factor: NMT 2.5 for the pseudoephedrine peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = chlorpheniramine peak response from the Sample solution rS = peak response from the Standard solution CS = concentration of USP Chlorpheniramine Maleate RS in the Standard solution (mg/mL) CU = nominal concentration of chlorpheniramine maleate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE Mobile phase, Pseudoephedrine hydrochloride stock solution, Pseudoephedrine hydrochloride solution, System suitability solution, and Chromatographic system: Proceed as directed in the Assay for Chlorpheniramine maleate. Standard stock solution: 0.6 mg/mL of USP Dextromethorphan Hydrobromide RS Standard solution 0.06 mg/mL of USP Dextromethorphan Hydrobromide RS in 0.1% phosphoric acid, from Standard stock solution rU
Acetaminophen, Chlorpheniramine Maleate, and Dextromethorphan Hydrobromide Tablets

(Comment on this Monograph)id=m2310=Acetaminophen, Chlorpheniramine Maleate, and Dextromethorphan Hydrobromide Tablets=A-Monos.pdf) DEFINITION Acetaminophen, Chlorpheniramine Maleate, and Detromethorphan Hydrobromide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), chlorpheniramine maleate (C16H19ClN2 C4H4O4), and dextromethorphan hydrobromide (C18H25NO HBr H2O). IDENTIFICATION A. The retention time of the major peak for acetaminophen from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. B. The retention time of the major peak for chlorpheniramine from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. C. The retention time of the major peak for dextromethorphan from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan hydrobromide. ASSAY ACETAMINOPHEN Mobile phase: Methanol, water, and glacial acetic acid (20:79:1) Standard solution: Transfer 50 mg of USP Acetaminophen RS to a 100-mL volumetric flask. Add 4 mL of methanol, and mix until solution is complete. Dilute with 0.1% phosphoric acid to volume. Sample stock solution: Transfer an equivalent to 100 mg of acetaminophen, from powdered Tablets (NLT 20), to a 50mL volumetric flask. Add 7.5 mL of methanol, and sonicate to disperse the powder. Add 0.5 mL of phosphoric acid, dilute with water to volume, and filter. Sample solution: Transfer 25.0 mL of the filtered Sample stock solution to a 100-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection volume: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 for the acetaminophen peak Relative standard deviation: NMT 2.0% from replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS = acetaminophen peak response from the Sample solution = peak response from the Standard solution
USP 32
Sample solution: Transfer an equivalent to 6 mg of dextromethorphan hydrobromide, from powdered Tablets (NLT 20), to a 100-mL volumetric flask. Add 10 mL of methanol, and sonicate to disperse the powder. Add 0.4 mL of phosphoric acid, and dilute with water to volume. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = dextromethorphan peak response from the Sample solution rS = peak response from the Standard solution CS = concentration of USP Dextromethorphan Hydrobromide RS in the Standard solution (mg/mL) CU = nominal concentration of dextromethorphan hydrobromide in the Sample solution (mg/mL) Mr1 = molecular weight of dextromethorphan hydrobromide monohydrate, 370.33 Mr2 = molecular weight of anhydrous dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Test 1 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Sample solution: Mix 9.0 mL of a filtered portion of the solution with 1.0 mL of 1% phosphoric acid solution Analysis: Determine the amounts of acetaminophen, chlorpheniramine maleate, and dextromethorphan hydrobromide dissolved, using the Analyses set forth in the Assay for Acetaminophen, Assay for Chlorpheniramine maleate, and Assay for Dextromethorphan hydrobromide, respectively, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2, C16H19ClN2 C4H4O4, and C18H25NO HBr H2O are dissolved. Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Medium: 0.1 M hydrochloric acid; 900 mL Apparatus, Time, Sample solution, Analysis, and Tolerances: Proceed as directed for Test 1. Test 3: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 3. Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL Apparatus, Time, Sample solution, Analysis, and Tolerances: Proceed as directed for Test 1. UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: The label states the name and quantity of each active ingredient and indicates its function (or purpose) in the article. When more than one Dissolution Test is given, the labeling states the Dissolution Test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Chlorpheniramine Maleate RS USP Dextromethorphan Hydrobromide RS USP Pseudoephedrine Hydrochloride RS
Acetaminophen and Codeine Phosphate Capsules

(Comment on this Monograph)id=m258=Acetaminophen and Codeine Phosphate Capsules=A-Monos.pdf) DEFINITION Acetaminophen and Codeine Phosphate Capsules contain NLT 90.0% and NMT 110.0% of the labeled quantities of acetaminophen (C8H9NO2) and codeine phosphate (C18H21NO3 H3PO4 1/2H2O). IDENTIFICATION A. The retention times of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 12 mg/mL each of USP Acetaminophen RS and USP Codeine Phosphate RS in methanol Sample solution: Transfer a portion of Capsule contents equivalent to 12 mg of codeine phosphate to a separator, and add 5 mL of water, 1 mL of ammonium hydroxide, and 5 mL of methylene chloride. Shake for 1 min, and allow the layers to separate. Use the clear, lower layer as the Sample solution. Developing solvent system: Methanol and ammonium hydroxide (49:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Analysis Samples: Standard solution and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF values of the two principal spots of the Sample solution correspond to those of the Standard solution. ASSAY PROCEDURE Solution A: Dissolve 2.04 g of monobasic potassium phosphate in 950 mL of water. Add 2 mL of triethylamine, adjust with phosphoric acid to a pH of 2.35, and dilute with water to 1000 mL. Mobile phase: Methanol and Solution A (8:92) Codeine phosphate standard stock solution: 0.3 mg/mL of USP Codeine Phosphate RS in Mobile phase Standard solution: Transfer 30 mg of USP Acetaminophen RS and 100J mL of Codeine phosphate standard stock solution (J being the ratio of the labeled amount, in mg, of codeine phosphate hemihydrate to that of acetaminophen) to a 100mL volumetric flask, dilute with Mobile phase to volume, and mix. This solution contains 0.3 mg/mL of acetaminophen and 0.3J mg/mL of codeine phosphate hemihydrate. Sample stock solution: Remove as completely as possible the contents of NLT 20 Capsules, weigh, and mix. Transfer a portion of the combined contents equivalent to 300 mg of acetaminophen to a 100-mL volumetric flask, add 75 mL of Mobile phase, and sonicate for 10 min. Dilute with Mobile phase to volume. Sample solution: Dilute 5.0 mL of the Sample stock solution with Mobile phase to 50 mL and pass a portion of the solution through a 1-m filter. Chromatographic system (See Chromatography 621, System Suitability.)
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rU rS CS
USP 32
= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) F = dilution factor, 1000 Calculate the quantity, in mg/mL, of C18H21NO3 H3PO4 1 /2H2O in the Capsule taken: Result = (rU/rS) CS (Mr1/Mr2) = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight of anhydrous codeine, 397.37 Mr2 Acceptance criteria: Meet the requirements rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Codeine Phosphate RS
Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 30 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between acetaminophen and codeine phosphate Relative standard deviation: NMT 2.0% and 3.0%, respectively, for acetaminophen and codeine phosphate Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the Capsules: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) CU = nominal concentration of acetaminophen in the Sample solution (mg/mL) Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in the Capsules: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of codeine phosphate CU hemihydrate in the Sample solution (mg/mL) Mr1 = molecular weight of codeine phosphate hemihydrate, 406.37 Mr2 = molecular weight of anhydrous codeine phosphate, 397.37 Acceptance criteria: 90.0%110.0% of the labeled amounts of C8H9NO2 and C18H21NO3 H3PO4 1/2H2O PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Analysis: Determine the amounts of C8H9NO2 and C18H21NO3 H3PO4 1/2H2O dissolved by using the method set forth in the Assay, except use 0.01 N hydrochloric acid to prepare the Codeine phosphate standard stock solution, and make any other necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled quantities of C8H9NO2 and C18H21NO3 H3PO4 1/2H2O is dissolved. UNIFORMITY OF DOSAGE UNITS 905 Procedure for content uniformity Solution A, Mobile phase, Codeine phosphate standard stock solution, Standard solution, and Chromatographic system: Prepare as directed in the Assay. Sample stock solution: Transfer the contents of 1 Capsule to a 100-mL volumetric flask, add 75 mL of Mobile phase, and sonicate for 10 min. Dilute with Mobile phase to volume. Sample solution: Dilute 5.0 mL of the Sample stock solution with Mobile phase to 50 mL and pass a portion of the solution through a 1-m filter. Analysis Samples: Standard solution and Sample solution Calculate the quantity, in mg, of C8H9NO2 in the Capsule taken: Result = (rU/rS) CS F rU rS CS rU rS CS
Acetaminophen and Codeine Phosphate Oral Solution

(Comment on this Monograph)id=m260=Acetaminophen and Codeine Phosphate Oral Solution=A-Monos.pdf) DEFINITION Acetaminophen and Codeine Phosphate Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled quantities of acetaminophen (C8H9NO2) and codeine phosphate hemihydrate (C18H21NO3 H3PO4 1/2H2O). IDENTIFICATION A. The retention times of the Sample solutions correspond to those of the Standard solutions, as obtained in the Assay for Acetaminophen and the Assay for Codeine phosphate. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 12 mg/mL each of USP Acetaminophen RS and USP Codeine Phosphate RS in methanol Sample solution: Transfer a volume of Oral Solution, equivalent to 12 mg of codeine phosphate, to a separator, add 1 mL of ammonium hydroxide, and 5 mL of methylene chloride, shake for 1 min, and allow the layers to separate. Use the clear, lower layer as the Sample solution. Developing solvent system: Methanol and ammonium hydroxide (49:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Analysis Samples: Standard solution and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF values of the two principal spots of the Sample solution correspond to those of the Standard solution.
USP 32
ASSAY ACETAMINOPHEN Mobile phase: Methanol and water (3:7) Standard solution: 0.48 mg/mL of USP Acetaminophen RS in Mobile phase Sample solution: Equivalent of 0.48 mg/mL of acetaminophen in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2.0 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates determined from the analyte peak Tailing factor: NMT 1.5 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = acetaminophen peak response from the Sample solution = peak response from the Standard solution rS CS = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) CU = nominal concentration of acetaminophen in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% CODEINE PHOSPHATE Mobile phase: Dissolve 4.44 g of docusate sodium in 1000 mL of a mixture of methanol, tetrahydrofuran, phosphoric acid, and water (600:40:1:360) with stirring, and pass through a membrane filter having a 0.45-m or finer porosity. Solution A: Methanol and water (3:7) Standard solution: 0.12 mg/mL of USP Codeine Phosphate RS in Solution A Sample solution: Equivalent of 0.12 mg/mL of acetaminophen in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates determined from the analyte peak Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in the Oral Solution taken: rU Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS = codeine phosphate peak response from the Sample solution = peak response from the Standard solution CS

= concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of codeine phosphate in CU the Sample solution (mg/mL) = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight of anhydrous codeine Mr2 phosphate, 397.37 Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 (for Oral Solution packaged in single-unit containers): Meets the requirements DELIVERABLE VOLUME 698 (for Oral Solution packaged in multiple-unit containers): Meets the requirements SPECIFIC TESTS PH 791: 4.06.1 ALCOHOL DETERMINATION, Method II 611 (test if alcohol is present): 90.0%120.0% of the labeled quantity of C2H5OH, acetone being used as the internal standard ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Codeine Phosphate RS
Acetaminophen and Codeine Phosphate Oral Suspension

(Comment on this Monograph)id=m265=Acetaminophen and Codeine Phosphate Oral Suspension=A-Monos.pdf) DEFINITION Acetaminophen and Codeine Phosphate Oral Suspension is a suspension of Acetaminophen and Codeine Phosphate in a suitable aqueous vehicle. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2) and codeine phosphate hemihydrate (C18H21NO3 H3PO4 1 /2H2O). IDENTIFICATION A. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 12 mg/mL each of USP Acetaminophen RS and USP Codeine Phosphate RS in methanol Sample solution: Transfer a volume of Oral Suspension, equivalent to 12 mg of codeine phosphate, to a separator, add 1 mL of ammonium hydroxide, and 5 mL of methylene chloride, shake for 1 min, and allow the layers to separate. Use the clear, lower layer as the Sample solution. Developing solvent system: Methanol and ammonium hydroxide (49:1) Chromatographic system (See Chromatography 621, Thin Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Analysis Samples: Standard solution and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Locate the spots on the plate by examination under short-wavelength UV light.
34

CU
USP 32
= nominal concentration of acetaminophen in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in the Oral Suspension taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of codeine phosphate CU hemihydrate in the Sample solution (mg/mL) = molecular weight ratio of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight ratio of anhydrous codeine Mr2 phosphate, 397.37 Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for Oral Suspension packaged in single-unit containers DELIVERABLE VOLUME 698: Meets the requirements for Oral Suspension packaged in multiple-unit containers SPECIFIC TESTS PH 791: 4.06.1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Codeine Phosphate RS rU rS CS
Acceptance criteria: The RF values of the two principal spots from the Sample solution correspond to those from the Standard solution. ASSAY PROCEDURE Mobile phase: Dissolve 4.9 g of monobasic potassium phosphate in 900 mL of water, adjust with phosphoric acid to a pH of 3.9, and add 216 mg of sodium 1octanesulfonate. Add 100 mL of acetonitrile. Solution A: Methanol and 0.01 N sodium hydroxide (30:70) Codeine phosphate standard stock solution: 0.5 mg/mL of USP Codeine Phosphate RS in Mobile phase Standard stock solution: Transfer a quantity of 5J mg of USP Acetaminophen RS, J being the ratio of the labeled amount, in mg, of acetaminophen to the labeled amount, in mg, of codeine phosphate hemihydrate, and 10.0 mL of Codeine phosphate standard stock solution to a 100-mL volumetric flask, and dissolve in and dilute with Solution A to volume. Standard solution: Dilute 10.0 mL of the Standard stock solution to 50 mL with Mobile phase (0.01 mg/mL of USP Codeine Phosphate RS and 0.01J mg/mL of USP Acetaminophen RS). Sample stock solution: Transfer a measured volume of well-mixed Oral Suspension, equivalent to 50 mg of acetaminophen, to a 100-mL volumetric flask. Add 50 mL of Solution A, and mix by mechanical means for 30 min. Dilute with Solution A to volume. [NOTEFoaming may be minimized by adding a few drops of acetonitrile before diluting with Solution A to volume.] Centrifuge a portion of this mixture. Sample solution: Dilute 10.0 mL of the clear supernatant from the Sample stock solution to 50 mL with Mobile phase. System suitability stock solution: 0.02 mg/mL of sodium benzoate and 0.03 mg/mL of methylparaben in Solution A System suitability solution: To 10.0 mL of the System suitability stock solution add 10.0 mL of Standard stock solution, and dilute to 50 mL with Mobile phase. Chromatographic system (See Chromatography 621,System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; 5-m packing L11 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: System suiltability solution and Standard solution [NOTEThe relative retention times for acetaminophen, benzoate, codeine, and methylparaben are about 0.25, 0.5, 1.0, and 1.3, respectively.] Suitability requirements Resolution: NLT 2 between each pair of adjacent peaks, System suitability solution Tailing factor: NMT 2 for each analyte peak, Standard solution Column efficiency: NLT 500 theoretical plates, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C8H9NO2 in the Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL)
Acetaminophen and Codeine Phosphate Tablets

(Comment on this Monograph)id=m270=Acetaminophen and Codeine Phosphate Tablets=A-Monos.pdf) DEFINITION Acetaminophen and Codeine Phosphate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9 NO2) and codeine phosphate hemihydrate (C18H21NO3 H3PO4 1/2H2O). IDENTIFICATION A. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 12 mg/mL each of USP Acetaminophen RS and USP Codeine Phosphate RS in methanol Sample solution: Transfer a quantity of finely powdered Tablets, equivalent to 12 mg of codeine phosphate, to a separator, add 5 mL of water, 1 mL of ammonium hydroxide, and 5 mL of methylene chloride, shake for 1 min, and allow the layers to separate. Use the clear, lower layer as the Sample solution. Developing solvent system: Methanol and ammonium hydroxide (49:1) Chromatographic system (See Chromatography 621, Thin Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture
USP 32
Application volume: 10 L Analysis Samples: Standard solution and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF values of the two principal spots from the Sample solution correspond to those from the Standard solution. ASSAY PROCEDURE Solution A: Dissolve 2.04 g of monobasic potassium phosphate in about 950 mL of water. Add 2 mL of triethylamine, adjust with phosphoric acid to a pH of 2.35, and dilute with water to 1000 mL. Mobile phase: Methanol and Solution A (8:92) Codeine phosphate standard stock solution: 0.3 mg/mL of USP Codeine Phosphate RS in Mobile phase Standard solution: Transfer 30 mg of USP Acetaminophen RS and 100 J mL of Codeine phosphate standard stock solution, J being the ratio of the labeled amount, in mg, of codeine phosphate hemihydrate to that of acetaminophen, to a 100-mL volumetric flask, and dilute with Mobile phase to volume. This solution contains 0.3 mg/mL of acetaminophen and 0.3J mg/mL of codeine phosphate hemihydrate. Sample stock solution: Transfer an equivalent to 300 mg of acetaminophen, from a portion of finely powdered Tablets (NLT 20), to a 100-mL volumetric flask, add 75 mL of Mobile phase, and sonicate for 10 min. Dilute with Mobile phase to volume. Sample solution: Dilute 5.0 mL of the Sample stock solution with Mobile phase to 50 mL, and pass a portion of the solution through a suitable 1-m filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 30 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between acetaminophen and codeine phosphate Relative standard deviation: NMT 2.0% and 3.0%, respectively, for acetaminophen and codeine phosphate Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C8H9NO2 in the Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) CU = nominal concentration of acetaminophen in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in the Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU = peak response from the Sample solution rU rS CS rS CS

= peak response from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of codeine phosphate CU hemihydrate in the Sample solution (mg/mL) = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 Mr2 = molecular weight of anhydrous codeine phosphate, 397.37 Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Analysis: Determine the amounts of C8H9NO2 and C18H21NO3 H3PO4 1/2H2O dissolved by employing the method set forth in the Assay, except to use 0.01 N hydrochloric acid to prepare the Codeine phosphate standard stock solution and to make any other necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2 and C18H21NO3 H3PO4 1/2H2O is dissolved. UNIFORMITY OF DOSAGE UNITS 905 Procedure for content uniformity Solution A, Mobile phase, Codeine phosphate standard stock solution, Standard solution, and Chromatographic system: Prepare as directed in the Assay. Sample stock solution: Transfer 1 Tablet to a 100-mL volumetric flask, add 75 mL of Mobile phase, and sonicate for 10 min. Dilute with Mobile phase to volume. Sample solution: Dilute 5.0 mL of the Sample stock solution with Mobile phase to 50 mL and pass a portion of the solution through a suitable 1-m filter. Analysis Samples: Standard solution and Sample solution Calculate the quantity, in mg, of C8H9NO2 in the Tablet taken: Result = (rU/rS) CS F = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) F = dilution factor for the Tablet taken, 1000 Calculate the quantity, in mg/mL, of C18H21NO3 H3PO4 1 /2H2O in the Tablet taken: rU rS CS Result = (rU/rS) CU (Mr1/Mr2) F = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) F = dilution factor for the Tablet taken, 1000 = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight of anhydrous codeine Mr2 phosphate, 397.37 Acceptance criteria: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Codeine Phosphate RS rU rS CU
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Acceptance criteria: 90.0%110.0% DEXTROMETHORPHAN HYDROBROMIDE Solution A: 6.8 mg/mL of monobasic potassium phosphate in water Mobile phase: Acetonitrile and Solution A (9:11) Standard solution: 0.1 mg/mL of USP Dextromethorphan Hydrobromide RS, 0.04 mg/mL USP Doxylamine Succinate RS, and 0.2 mg/mL USP Pseudoephedrine Hydrochloride RS in Mobile phase Sample solution: Equivalent to 0.1 mg/mL of dextromethorphan hydrobromide from a volume of Oral Solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L9 Flow rate: 2.5 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for pseudoephedrine, dextromethorphan, and doxylamine are 0.38, 0.65, and 1.0, respectively.] Suitability requirements Column efficiency: NLT 500 theoretical plates for the dextromethorphan, doxylamine, and pseudoephedrine peaks Tailing factor: NMT 2.5 for the dextromethorphan, doxylamine, and pseudoephedrine peaks Relative standard deviation: NMT 2.0% for dextromethorphan, doxylamine, and pseudoephedrine Analysis Samples: Standard solution and Sample solution Calculate the quantity, in percentage, of C18H25NO HBr H2O in the portion of Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dextromethorphan Hydrobromide RS in the Standard solution (mg/mL) = concentration of dextromethorphan CU hydrobromide in the Sample solution (mg/mL) = molecular weight of dextromethorphan Mr1 hydrobromide monohydrate, 370.33 = molecular weight of anhydrous Mr2 dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% DOXYLAMINE SUCCINATE [NOTESolution A, Mobile phase, Standard solution, and Chromatographic system: proceed as directed in the Assay for Dextromethorphan Hydrobromide.] Sample solution: Equivalent to 0.04 mg/mL of doxylamine succinate from a volume of Oral Solution in Mobile phase Analysis: Proceed as directed for Procedure in the Assay for Dextromethorphan Hydrobromide Calculate the quantity, in percentage, of C17H22N2O C4H6O4 in the portion of Oral Solution taken: rU rS CS Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Doxylamine Succinate RS in the Standard solution (mg/mL)
Acetaminophen, Dextromethorphan Hydrobromide, Doxylamine Succinate, and Pseudoephedrine Hydrochloride Oral Solution
(Comment on this Monograph)id=m275=Acetaminophen, Dextromethorphan Hydrobromide, Doxylamine Succinate, and Pseudoephedrine Hydrochloride Oral Solution=A-Monos.pdf) DEFINITION Acetaminophen, Dextromethorphan Hydrobromide, Doxylamine Succinate, and Pseudoephedrine Hydrochloride Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amounts of Acetaminophen (C8H9NO2), Dextromethorphan Hydrobromide (C18H25NO HBr H2O), Doxylamine Succinate (C17H22N2O C4H6O4), and Pseudoephedrine Hydrochloride (C10H15NO HCl). IDENTIFICATION A. The retention time of the major peak for acetaminophen from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Acetaminophen. B. The retention time of the major peak for dextromethorphan from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Dextromethorphan Hydrobromide. C. The retention time of the major peak for doxylamine from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Doxylamine Succinate. D. The retention time of the major peak for pseudoephedrine from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Pseudoephedrine Hydrochloride. ASSAY ACETAMINOPHEN Mobile phase: Methanol and water (11:9) Standard solution: 0.2 mg/mL of USP Acetaminophen RS in Mobile phase Sample solution: Equivalent to 0.2 mg/mL of Acetaminophen from a volume of Oral Solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 500 theoretical plates determined from the analyte peak Tailing factor: NMT 2.0 for the acetaminophen peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample Solution Calculate the quantity, in percentage, of C8H9NO2 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (mg/mL) = concentration of acetaminophen in the Sample solution (mg/mL)
USP 32
= concentration of doxylamine succinate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE HYDROCHLORIDE [NOTESolution A, Mobile phase, Standard solution, and Chromatographic system: proceed as directed in the Assay for Dextromethorphan Hydrobromide.] Sample solution: Equivalent to 0.2 mg/mL of pseudoephedrine hydrochloride from a volume of Oral Solution in Mobile phase Analysis: Proceed as directed for Procedure in the Assay for Dextromethorphan Hydrobromide. Calculate the quantity, in percentage, of C10H15NO HCl in the portion of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Pseudoephedrine Hydrochloride RS in the Standard solution (mg/mL) = concentration of pseudoephedrine hydrochloride CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For oral solution packaged in single-unit containers, it meets the requirements. DELIVERABLE VOLUME 698: For oral solution packaged in multiple-unit containers, it meets the requirements. SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: The total bacterial count does not exceed 100 cfu/g, the total combined molds and yeasts count does not exceed 10 cfu/g, and it meets the requirements of the tests for absence of Salmonella species and Escherichia coli. PH 791: 4.56.3 ALCOHOL DETERMINATION, Method II (if present) 611: 90.0%110.0% of the labeled amount of C2H5OH ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Dextromethorphan Hydrobromide RS USP Doxylamine Succinate RS USP Pseudoephedrine Hydrochloride RS CU

ASSAY ACETAMINOPHEN Mobile phase: Methanol and water (2:3) Internal standard solution: 8.0 mg/mL of guaifenesin in a mixture of water and methanol (4:1) Standard solution: Transfer 50 mg of USP Acetaminophen RS to a 100-mL volumetric flask. Dissolve in 2.5 mL of methanol, and dilute with water to volume. Transfer 2.0 mL of this solution to a 50-mL volumetric flask, add 5.0 mL of Internal standard solution, and dilute with Mobile phase to volume. Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 500 mg of acetaminophen, to a 100-mL volumetric flask, add 25 mL of methanol, and shake by mechanical means for 10 min. Dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 2.0 mL of this solution to a 50-mL volumetric flask, add 5.0 mL of Internal standard solution, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Column temperature: 35 0.5 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen and guaifenesin are 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 6.0 between the analyte and internal standard peaks Column efficiency: NLT 1000 theoretical plates determined from the analyte peak Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2.5% of the peak response ratios for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of acetaminophen to the internal standard peak from the Sample solution = peak response ratio of acetaminophen to the RS internal standard peak from the Standard solution = concentration of USP Acetaminophen RS in the CS Standard solution (mg/mL) = nominal concentration of acetaminophen in the CU Sample solution (mg/mL) DIPHENHYDRAMINE CITRATE Mobile phase: Methanol, water, and glacial acetic acid (61:38:1) containing 1.0813 g of sodium 1-octanesulfonate in each 1000 mL of solution Solution A: Methanol and water (1:1) Internal standard solution: 8.0 mg/mL of xylometazoline hydrochloride Standard solution: Transfer 38 mg of USP Diphenhydramine Citrate RS to a 100-mL volumetric flask containing 500 mg of acetaminophen. Add 5.0 mL of Internal standard solution and 50 mL of Solution A, and mix until solution is complete. Dilute with Solution A to volume. Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 38 mg of diphenhydramine citrate, to a 100-mL volumetric flask, add RU
Acetaminophen and Diphenhydramine Citrate Tablets

(Comment on this Monograph)id=m280=Acetaminophen and Diphenhydramine Citrate Tablets=A-Monos.pdf) DEFINITION Acetaminophen and Diphenhydramine Citrate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2) and diphenhydramine citrate (C17H21NO C6H8O7). IDENTIFICATION The retention times of the major peaks of the Sample solutions, obtained in the Assay for Acetaminophen and in the Assay for Diphenhydramine Citrate, relative to the retention times of the respective internal standards, correspond to those of the respective Standard solutions.
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DEFINITION Acetaminophen, Diphenhydramine Hydrochloride, and Pseudoephedrine Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2), diphenhydramine hydrochloride (C17H21NO HCl), and pseudoephedrine hydrochloride (C10H15NO HCl). IDENTIFICATION A. The retention time of the major peak for acetaminophen in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. B. The retention time of the major peak for diphenhydramine in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. C. The retention time of the major peak for pseudoephedrine in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. ASSAY ACETAMINOPHEN Solution A: Transfer 6.8 g of monobasic potassium phosphate to a 1000-mL volumetric flask, and add water to dissolve. Add 2.0 mL of triethylamine, and dilute with water to volume. Adjust with phosphoric acid to a pH of 4.0. Diluent: Acetonitrile and Solution A (11:89) Mobile phase: Acetonitrile and Solution A (3:47) Standard solution: 25 g/mL of USP Acetaminophen RS, 12.5 g/mL of USP Diphenhydramine Hydrochloride RS, and 30 g/mL of USP Pseudoephedrine Hydrochloride RS in Diluent Sample stock solution: Weigh and finely powder NLT 20 Tablets, and transfer a portion of the powder, equivalent to 500 mg of acetaminophen, to a 100-mL volumetric flask. Add 75 mL of Diluent, shake, and sonicate for 15 min. Cool to room temperature, dilute with Diluent to volume, and mix. Sample solution: Dilute an accurately measured volume of the Sample stock solution, stepwise if necessary, with Diluent to obtain a solution having a concentration of 25 g/mL of acetaminophen. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L10 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3000 theoretical plates determined from the acetaminophen, diphenhydramine, and pseudoephedrine peaks Tailing factor: NMT 2.0 for the acetaminophen, diphenhydramine, and pseudoephedrine peaks Relative standard deviation: NMT 2.0% determined from the acetaminophen, diphenhydramine hydrochloride, and pseudoephedrine hydrochloride responses for replicate injections Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C8H9NO2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetaminophen RS in the Standard solution (g/mL)
65 mL of Solution A, and shake by mechanical means for 15 min. Add 5.0 mL of Internal standard solution, and dilute with Solution A to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 3.9-mm 30-cm; packing L1 Column temperature: 35 0.5 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen, diphenhydramine citrate, and xylometazoline hydrochloride are 0.3, 0.7, and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between the analyte and internal standard peaks Column efficiency: NLT 1000 theoretical plates determined from the analyte peak Tailing factor: NMT 1.7 for the analyte peak Relative standard deviation: NMT 2.0% of the peak response ratios for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H21NO C6H8O7 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Diphenhydramine Citrate RS in the Standard solution (mg/mL) = nominal concentration of diphenhydramine CU citrate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711, Procedure for a Pooled Sample Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Analysis: Determine the amount of C8H9NO2 and of C17H21NO C6H8O7 dissolved, using the Analysis set forth in the Assay for Acetaminophen and the Assay for Diphenhydramine Citrate, respectively, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2 and C17H21NO C6H8O7 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to acetaminophen and to diphenhydramine citrate ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Diphenhydramine Citrate RS RU RS CS
Acetaminophen, Diphenhydramine Hydrochloride, and Pseudoephedrine Hydrochloride Tablets

(Comment on this Monograph)id=m285=Acetaminophen, Diphenhydramine Hydrochloride, and Pseudoephedrine Hydrochloride Tablets=A-Monos.pdf)
USP 32
= nominal concentration of acetaminophen in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% DIPHENHYDRAMINE HYDROCHLORIDE Solution A, Diluent, Mobile phase, Standard solution, and Chromatographic system: Proceed as directed in the Assay for Acetaminophen. Sample stock solution: Weigh and finely powder NLT 20 Tablets, and transfer a portion of the powder, equivalent to 12.5 mg of diphenhydramine hydrochloride, to a 100-mL volumetric flask. Add 75 mL of Diluent, shake, and sonicate for 15 min. Cool to room temperature, dilute with Diluent to volume, and mix. Sample solution: Dilute a volume of Sample stock solution, stepwise if necessary, with Diluent to obtain a solution having a concentration of 12.5 g/mL of diphenhydramine. Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C17H21NO HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Diphenhydramine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of diphenhydramine CU hydrochloride in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE HYDROCHLORIDE Solution A, Diluent, Mobile phase, Standard solution, and Chromatographic system: Proceed as directed in the Assay for Acetaminophen. Sample stock solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 30 mg of pseudoephedrine hydrochloride, to a 100-mL volumetric flask, add about 75 mL of Diluent, shake, and sonicate for 15 min. Cool to room temperature, and dilute with Diluent to volume. Sample solution: Dilute a volume of Sample stock solution, stepwise if necessary, with Diluent to obtain a solution having a concentration of 30 g/mL of pseudoephedrine hydrochloride. Calculate the percentage of C10H15NO HCl in the portion of Tablets taken: rU rS CS Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Pseudoephedrine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of pseudoephedrine CU hydrochloride in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL Apparatus 2: 50 rpm Time: 45 min Analysis: Determine the amounts of C8H9NO2, C17H21NO HCl, and C10H15NO HCl dissolved by employing the following method. Solution A, Diluent, Standard solution, Mobile phase, and Chromatographic system: Proceed as directed in the Assay for Acetaminophen. CU

Sample solution A: Combine equal volumes of the filtered solutions, and use the pooled sample. Sample solution B: 5.0 mL of Sample solution A to a 100mL volumetric flask Dilute with Mobile phase to volume. Analysis: Using Sample solution A and the Standard solution, and making any necessary volumetric adjustments, proceed as directed in the Assay for Diphenhydramine hydrochloride and the Assay for Pseudoephedrine Hydrochloride, and determine the amounts of C17H21NO HCl and C10H15NO HCl dissolved. Using Sample solution B and the Standard solution, and making any necessary volumetric adjustments, proceed as directed in the Assay for Acetaminophen, and determine the amount of C8H9NO2 dissolved. Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2, C17H21NO HCl, and C10H15NO HCl is dissolved. For tablets labeled as chewable: Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL Apparatus 2: 75 rpm Time: 45 min Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2, C17H21NO HCl and C10H15NO HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Diphenhydramine Hydrochloride RS USP Pseudoephedrine Hydrochloride RS
Acetaminophen and Pseudoephedrine Hydrochloride Tablets

(Comment on this Monograph)id=m290=Acetaminophen and Pseudoephedrine Hydrochloride Tablets=A-Monos.pdf) DEFINITION Acetaminophen and Pseudoephedrine Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of acetaminophen (C8H9NO2) and pseudoephedrine hydrochloride (C10H15NO HCl). IDENTIFICATION The retention times of the acetaminophen and pseudoephedrine peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Acetonitrile and water (1:9) Solution A: 0.005 M ethanesulfonic acid and 0.05 M monobasic potassium phosphate Mobile phase: Acetonitrile and Solution A (1:9) Adjust with 5 N sodium hydroxide or 1 N hydrochloric acid to a pH of 4.6. Pseudoephedrine hydrochloride standard stock solution: USP Pseudoephedrine Hydrochloride RS in Diluent (0.6 mg/mL) Standard solution: Transfer 6J mg of USP Acetaminophen RS to a 100-mL volumetric flask, J being the ratio of the labeled quantity (mg) of acetaminophen to the labeled quantity (mg) of pseudoephedrine hydrochloride in each Tablet. Add 2.0 mL of 1 N hydrochloric acid and 20 mL of Diluent, and mix to dissolve. Add 10.0 mL of Pseudoephedrine hydrochloride stock standard solution, and dilute with Diluent to volume. This solution contains 0.06J
40
USP 32
Calculate the percentage of C8H9NO2 and C10H15NO HCl dissolved: Result = (rU/rS) V (CS/L) 100 = peak response of the corresponding analyte of the Sample solution = peak response of the corresponding analyte of rS the Standard solution V = volume of Medium, 900 mL = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) L = label amount of the corresponding analyte in a Tablet (mg) Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2 and C10H15NO HCl is dissolved. For tablets labeled as chewable Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL Apparatus 2: 75 rpm Time: 45 min Standard solution, Sample solution, Chromatographic system, and Analysis: Proceed as directed above in Procedure for a Pooled Sample. Tolerances: NLT 75% (Q) of the labeled amounts of C8H9NO2 and C10H15NO HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Pseudoephedrine Hydrochloride RS
mg/mL of USP Acetaminophen RS and 0.06 mg/mL of USP Pseudoephedrine Hydrochloride RS. Sample solution: Transfer an equivalent to 30 mg of pseudoephedrine hydrochloride, from finely powdered tablets (NLT 20), to a 500-mL volumetric flask, add 10.0 mL of 1 N hydrochloric acid and 100 mL of Diluent, and sonicate for 30 min, with occasional shaking. Allow to cool, and dilute with Diluent to volume. Pass a portion of this solution through a glass fiber filter, and use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; base-deactivated or endcapped packing L1 Flow rate: 3 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen and pseudoephedrine are about 0.55 and 1.0, respectively. The retention time for acetaminophen is NLT 2 min.] Suitability requirements Resolution: NLT 3.5 between acetaminophen and pseudoephedrine Tailing factor: NMT 2.0 for the pseudoephedrine peak Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO2 and C10H15NO HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response for the corresponding analyte of the Sample solution rS = peak response for the corresponding analyte of the Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of the appropriate analyte CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: pH 5.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL Apparatus 2: 50 rpm Time: 45 min Determine the amount of C8H9NO2 and C10H15NO HCl dissolved by employing the following method. Mobile phase: Proceed as directed in the Assay. Standard solution: L/900 mg/mL of USP Pseudoephedrine Hydrochloride RS and LJ/900 mg/mL of USP Acetaminophen RS in Medium [NOTEL is the labeled quantity, in mg, of pseudoephedrine hydrochloride in each Tablet; and J is the ratio of the labeled quantity, in mg, of acetaminophen to the labeled quantity, in mg, of pseudoephedrine hydrochloride in each Tablet.] Sample solution: Sample per Dissolution 711; filter. Chromatographic system and System suitability: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution [NOTEInject 20 L of the Samples, and measure the responses for the acetaminophen and pseudoephedrine peaks.] rU
Acetazolamide
(Comment on this Monograph)id=m320=Acetazolamide=AMonos.pdf)
222.25 C4H6N4O3S2 Acetamide, N-[5-(aminosulfonyl)-1,3,4-thiadiazol-2-yl]-; N-(5-Sulfamoyl-1,3,4-thiadiazol-2-yl)acetamide [59-66-5]. DEFINITION Acetazolamide contains NLT 98.0% and NMT 102.0% of C4H6N4O3S2, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solution: 20 mg/mL in 1 N sodium hydroxide Analysis: To 5 mL of Sample solution, add 5 mL of a solution made by dissolving 100 mg of hydroxylamine hydrochloride and 80 mg of cupric sulfate in 10 mL of water. Heat the resulting pale yellow solution on a steam bath for 5 min. Acceptance criteria: A clear, bright yellow solution is produced. No heavy precipitate or dark brown color results after the mixing or heating.
USP 32
ASSAY PROCEDURE Standard solution: 20 mg/mL of USP Acetazolamide RS in pyridine Sample solution: 20 mg/mL of Acetazolamide in pyridine Blank: Pyridine Spectrometric conditions Mode: IR Analytical wavelength: 7.38 m (1350 cm 1) Cell: 0.1 mm Analysis Samples: Standard solution and Sample Solution Calculate the quantity, as a percentage, of C4H6N4O3S2 in the portion taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Acetazolamide RS in the Standard solution (mg/mL) = concentration of Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% AU AS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221: Digest 1.5 g with 75 mL of water at about 70 for 5 min. Cool to room temperature, and filter. Acceptance criteria: A 25-mL portion of the filtrate shows no more chloride than corresponds to 0.10 mL of 0.020 N hydrochloric acid (0.014%) CHLORIDE AND SULFATE, Sulfate 221: A 25-mL portion of the filtrate prepared in the test for Chloride shows no more sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid (0.04%). SELENIUM 291: 30 ppm, a 200-mg specimen being used HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Standard solution and Sample solution: Acetone and methanol (1:1) Eluant: n-Propyl alcohol and 1 N ammonium hydroxide (22:3) Visualization: 1 SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5% SILVER-REDUCING SUBSTANCES Sample: 5 g Analysis: Thoroughly wet the Sample with alcohol. Add 125 mL of water, 10 mL of nitric acid, and 5.0 mL of 0.1 N silver nitrate VS. Stir with a mechanical stirrer for 30 min. Filter, add 5 mL of ferric ammonium sulfate TS to the filtrate, and titrate with 0.1 N ammonium thiocyanate VS to a reddishbrown endpoint. Acceptance criteria: NLT 4.8 mL of 0.1 N ammonium thiocyanate is required, calculated on the anydrous basis. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at room temperature. USP REFERENCE STANDARDS 11 USP Acetazolamide RS
Official Monographs / Acetazolamide 41
Acetazolamide for Injection

(Comment on this Monograph)id=m350=Acetazolamide for Injection=A-Monos.pdf) DEFINITION Acetazolamide for Injection is prepared from Acetazolamide with the aid of Sodium Hydroxide. It is suitable for parenteral use. The contents of each container, when constituted as directed in the labeling, yield a solution containing NLT 95.0% and NMT 110.0% of the labeled amount of acetazolamide (C4H6N4O3S2). IDENTIFICATION A. PROCEDURE Sample: Dissolve 500 mg in 5 mL of water, add 2 drops of hydrochloric acid, and allow the mixture to stand for about 15 min. Filter through a fine sintered-glass funnel, wash with several small portions of water, and dry in vacuum over silica gel for 3 h: the crystals meet the requirements of the following tests. Analysis 1: Infrared Absorption 197K Analysis 2 Sample solution: 100 mg/mL of the Sample in 1N sodium hydroxide To 5 mL of the Sample solution, add 5 mL of a solution made by dissolving 100 mg of hydroxylamine hydrochloride and 80 mg of cupric sulfate in 10 mL of water. Mix, and heat the resulting pale yellow solution on a steam bath for 5 min. Acceptance criteria: A clear, bright yellow solution is produced. No heavy precipitate or dark brown color results after the mixing or heating. B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements ASSAY PROCEDURE Standard stock solution: 100 g/mL of USP Acetazolamide RS in sodium hydroxide solution (1 in 100) Standard solution: Standard stock solution and 0.1 N hydrochloric acid (1:9) Sample stock solution: 500 g/mL of acetazolamide from Acetazolamide for Injection, in water [NOTEDissolve the contents of 1 container of Acetazolamide for Injection in a measured volume of water corresponding to the volume of solvent specified in the labeling. Dilute a portion of this solution to obtain the Sample stock solution.] Sample solution: Sample stock solution, 1 N hydrochloric acid, and water (1:5:44) Blank: 0.1 N hydrochloric acid Spectrometric conditions Mode: UV Analytical wavelength: 265 nm Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C4H6N4O3S2 in the portion of Acetazolamide for Injection taken: Result = (AU/AS) (CS/CU) 100 AU AS = absorbance of the Sample solution = absorbance of the Standard solution
42
Acetazolamide / Official Monographs

CS
USP 32
Sample solution: 250 g/mL of acetazolamide from Acetazolamide Oral Suspension in Mobile phase [NOTEAgitate the container of Oral Suspension for 30 min on a rotating mixer, remove a 5-mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the Sample solution from the freezer, allow to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 1.0 mL of the Sample solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention time for the acetazolamide peak is about 3 min.] Suitability requirements Relative standard deviation: Acetazolamide, NMT 1.1% for replicate injections of the Standard solution Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C4H6N4O3S2 in the portion of Acetazolamide Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acetazolamide RS in the Standard solution (g/mL) CU = nominal concentration of acetazolamide in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.05.0 (Vehicle for Oral Solution and Vehicle for Oral Suspension), and 3.13.9 (Cherry Syrup) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at controlled room temperature, or in a cold place. LABELING: Label it to state that it is to be well shaken before use, and to state the beyond-use date. BEYOND-USE DATE: 60 days after the day on which it was compounded USP REFERENCE STANDARDS 11 USP Acetazolamide RS rU rS CS
= concentration of USP Acetazolamide RS in the Standard solution (g/mL) = nominal concentration of acetazolamide in the CU Sample solution (g/mL) Acceptance criteria: 95.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
SPECIFIC TESTS PH 791: 9.010.0, in a freshly prepared solution (1 in 10) BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.5 USP Endotoxin Unit/mg of acetazolamide. INJECTIONS 1, Constituted Solutions: At the time of use, it meets the requirements. STERILITY TESTS 71: It meets the requirements. COMPLETENESS OF SOLUTION 641: 100 mg/mL in carbon dioxidefree water dissolves to yield a clear solution. INJECTIONS 1, Labeling: It meets the requirements. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids, preferably of Type III glass, and store at room temperature. USP REFERENCE STANDARDS 11 USP Acetazolamide RS USP Endotoxin RS
Acetazolamide Oral Suspension

(Comment on this Monograph)id=m1376=Acetazolamide Oral Suspension=A-Monos.pdf) DEFINITION Acetazolamide Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of acetazolamide (C4H6N4O3S2). Prepare Acetazolamide Oral Suspension, 25 mg/mL, as follows. (See Pharmaceutical CompoundingNonsterile Preparations 795.) (See also Acetazolamide Oral Solution.)
Acetazolamide Vehicle: a mixture of Vehicle for Oral Solution, NF (regular or sugar-free), and Vehicle for Oral Suspension, NF (1:1), or Cherry Syrup, NF To make 2.5 g A sufficient quantity
100 mL
preparation becomes a suspension and should be labeled as such.] If using Tablets, place the Tablets in a mortar and comminute the Tablets to a fine powder, or add Acetazolamide powder. Add about 20 mL of the Vehicle, and mix to a uniform paste. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add enough liquid Vehicle to bring to final volume, and mix well. ASSAY PROCEDURE Mobile phase: Dissolve 4.1 g of anhydrous sodium acetate in 950 mL of water, and add 20 mL of methanol and 30 mL of acetonitrile. Adjust with glacial acetic acid to a pH of 4.0. Standard stock solution: 0.5 mg/mL of USP Acetazolamide RS in 0.5 N sodium hydroxide solution and water (1.0:9.0) Standard solution: 250 g/mL from Standard stock solution in water
[NOTEIf Tablets are used instead of bulk powder, the
Acetazolamide Tablets
(Comment on this Monograph)id=m360=Acetazolamide Tablets=A-Monos.pdf) DEFINITION Acetazolamide Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of acetazolamide (C4H6N4O3S2). IDENTIFICATION Sample stock solution: Equivalent to 10 mg/mL of acetazolamide in acetone Sample: Filter the Sample stock solution, and add sufficient solvent hexane to the filtrate to cause formation of a heavy,
USP 32
white precipitate. Collect the precipitate on a mediumporosity, sintered-glass funnel, and dry with suction. A. INFRARED ABSORPTION 197K: Use the Sample. B. PROCEDURE: Dissolve 100 mg of Sample in 5 mL of 1 N sodium hydroxide, add 5 mL of a solution made by dissolving 100 mg of hydroxylamine hydrochloride and 80 mg of cupric sulfate in 10 mL of water. Heat the resulting pale yellow solution on a steam bath for 5 min: a clear, bright yellow solution is produced. Acceptance criteria: No heavy precipitate or dark brown color results after the mixing or heating. ASSAY PROCEDURE Mobile phase: 4.1 g of anhydrous sodium acetate in 950 mL. Add 20 mL of methanol and 30 mL of acetonitrile. Adjust with glacial acetic acid to a pH of 4.0 0.05. Internal standard stock solution: 10 mg/mL of sulfadiazine in 0.5 N sodium hydroxide Internal standard solution: 1 mg/mL from Internal standard stock solution in water Standard stock solution: Dissolve 25 mg of USP Acetazolamide RS in 2.5 mL of 0.5 N sodium hydroxide. Dilute with water to 25 mL. Standard solution: Transfer 10.0 mL of Standard acetazolamide stock solution, 10.0 mL of Internal standard solution, and 10 mL of 0.5 N sodium hydroxide to a 100-mL volumetric flask. Dilute with water to volume. Sample stock solution: Transfer a portion of the powder equivalent to 100 mg acetazolamide. Add 10 mL of 0.5 N sodium hydroxide, sonicate for 5 min, cool to room temperature, and dilute with water to 100 mL. Filter a portion of this solution, discarding the first 20 mL of the filtrate. Sample solution: Transfer 10.0 mL of Sample stock solution, 10.0 mL of Internal standard solution, and 10 mL of 0.5 N sodium hydroxide to a 100-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetazolamide and sulfadiazine are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the analyte and internal standard peaks Relative standard deviation: NMT 1.0% for the ratios of the analyte peak response to the internal standard peak response Analysis Samples: Standard solution and Sample solution Calculate the quantity, as a percentage, of C4H6N4O3S2 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = peak response ratios of the analyte peak to the internal standard peak obtained from the Sample solution = peak response ratios of the analyte peak to the internal standard peak obtained from the Standard solution = concentration of USP Acetazolamide RS in Standard solution (mg/mL) = nominal concentration of acetazolamide in the Sample solution (mg/mL) Acceptance criteria:
Official Monographs / Glacial 43

95.0%105.0%
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 60 min Detector: UV 265 nm Standard solution: USP Acetazolamide RS in the same Medium Sample solutions: Sample per Dissolution 711. Analysis: Determine the amount of C4H6N4O3S2 dissolved by using UV absorption on filtered portions of the Sample solution suitably diluted with Medium, if necessary, in comparison with a Standard solution having a known concentration of USP Acetazolamide RS. Acceptance criteria: NLT 75% (Q) of the labeled amount of C4H6N4O3S2 is dissolved UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetazolamide RS
Glacial Acetic Acid

(Comment on this Monograph)id=m440=Glacial Acetic Acid=AMonos.pdf)
C2H4O2 Acetic acid [64-19-7].
60.05
DEFINITION Glacial Acetic Acid contains NLT 99.5% and NMT 100.5%, by weight, of C2H4O2. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Acetate 191: Meets the requirements Sample solution: Glacial Acetic Acid and water (1:2) ASSAY PROCEDURE Sample solution: Measure 2 mL of Glacial Acetic Acid into a glass-stoppered flask, previously tared while containing about 20 mL of water, and weigh again to obtain the weight of the substance under assay. Analysis: Add 20 mL of water, then add phenolphthalein TS. Titrate with 1 N sodium hydroxide VS. Each mL of 1 N sodium hydroxide is equivalent to 60.05 mg of C2H4O2. Acceptance criteria: 99.5%100.5% IMPURITIES Inorganic Impurities LIMIT OF NONVOLATILE RESIDUE: Evaporate 20 mL in a tared dish, and dry at 105 for 1 h: the weight of the residue does not exceed 1.0 mg. HEAVY METALS 231: NMT 5 ppm Sample solution: To the residue obtained in the test for Limit of Nonvolatile Residue add 8 mL of 0.1 N hydrochloric acid, warm gently until solution is complete, dilute with water to 100 mL, and use 20 mL.
44
Glacial / Official Monographs
USP 32
CHLORIDE AND SULFATE, Chloride 221 Sample solution: Dilute 1.0 mL with 20 mL of water. Analysis: Add 5 drops of silver nitrate TS. Acceptance criteria: No opalescence is produced. CHLORIDE AND SULFATE, Sulfate 221 Sample: Dilute 1.0 mL with 10 mL of water. Analysis: Add 1 mL of barium chloride TS. Acceptance criteria: No turbidity is produced. Organic Impurities PROCEDURE: READILY OXIDIZABLE SUBSTANCES Sample solution: Dilute 2.0 mL in a glass-stoppered vessel with 10 mL of water. Analysis: Add 0.10 mL of 0.10 N potassium permanganate. Acceptance criteria: The pink color is not changed to brown within 2 h. SPECIFIC TESTS CONGEALING TEMPERATURE 651: NLT 15.6
Acetic Acid Otic Solution

(Comment on this Monograph)id=m460=Acetic Acid Otic Solution=A-Monos.pdf) DEFINITION Acetic Acid Otic Solution is a solution of Glacial Acetic Acid in a suitable nonaqueous solvent. It contains NLT 85.0% and NMT 130.0% of the labeled amount of C2H4O2. IDENTIFICATION A. PROCEDURE Sample solution: Acetic Acid Otic Solution and water (1:2) Analysis: Adjust 10 mL of the Sample solution with 1 N sodium hydroxide to a pH of 7. Add ferric chloride TS. Acceptance criteria: A deep red color is produced, and it is destroyed by the addition of hydrochloric acid. B. Warm it with sulfuric acid and alcohol: ethyl acetate, recognizable by its characteristic odor, is evolved. ASSAY PROCEDURE Sample solution: Transfer a quantity of Otic Solution, containing 100 mg of glacial acetic acid, to a 250-mL conical flask, and add 5 mL of saturated sodium chloride solution and 40 mL of water. Add 3 drops of phenolphthalein TS. Analysis: Titrate with 0.1 N sodium hydroxide VS to a faint pink endpoint. Each mL of 0.1 N sodium hydroxide is equivalent to 6.005 mg of C2H4O2. Acceptance criteria: 85.0%130.0% SPECIFIC TESTS PH 791: 2.04.0, when diluted with an equal volume of water ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at room temperature.
Acetic Acid Irrigation

(Comment on this Monograph)id=m450=Acetic Acid Irrigation=A-Monos.pdf) DEFINITION Acetic Acid Irrigation is a sterile solution of Glacial Acetic Acid in Water for Injection. It contains, in each 100 mL, NLT 237.5 mg and NMT 262.5 mg of C2H4O2. IDENTIFICATION PROCEDURE Evaporate 100 mL to about 10 mL: the resulting solution meets the requirements for Identification TestsGeneral, Acetate 191. ASSAY PROCEDURE Analysis: Pipet 50 mL of Irrigation into a 150-mL conical flask, add 2 drops of phenolphthalein TS, and titrate with 0.1 N sodium hydroxide VS. Each mL of 0.1 N sodium hydroxide is equivalent to 6.005 mg of C2H4O2. Acceptance criteria: 237.5262.5 mg of C2H4O2 SPECIFIC TESTS PH 791: 2.83.4 BACTERIAL ENDOTOXINS TEST 85 It contains NMT 0.5 Endotoxin Unit/mL. OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that the container in which it is packaged may be designed to empty rapidly and may exceed 1000 mL in capacity. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I or Type II glass, and store at controlled room temperature. It may be packaged in suitable plastic containers. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Acetohexamide
(Comment on this Monograph)id=m510=Acetohexamide=AMonos.pdf)
C15H20N2O4S 324.40 Benzenesulfonamide, 4-acetyl-N-[[cyclohexylamino]carbonyl]-; 1-[(p-Acetylphenyl)sulfonyl]-3-cyclohexylurea [968-81-0]. DEFINITION Acetohexamide contains NLT 97.0% and NMT 101.0% of C15H20N2O4S, calculated on the dried basis.
USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Solution: 10 g/mL in 0.01 N sodium hydroxide Analytical wavelength: 247 nm Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0%. ASSAY PROCEDURE Sample solution: Dissolve about 300 mg of Acetohexamide in 40 mL of dimethylformamide. Analysis: To the Sample solution, add 5 drops of thymol blue TS, and titrate, using a magnetic stirrer, with 0.1 N sodium methoxide VS to a blue endpoint. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N sodium methoxide is equivalent to 32.44 mg of C15H20N2O4S. Acceptance criteria: 97.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% SELENIUM 291: 30 ppm; a 200-mg specimen mixed with 200 mg of magnesium oxide being used HEAVY METALS, Method II 231: NMT 20 ppm SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 182.2187 LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Acetohexamide RS
Official Monographs / Acetohydroxamic 45

Sample solution: Quantitatively dilute Sample stock solution 3 (1 in 10) in water (10 g/mL). Blank: 0.01 N sodium hydroxide Spectrometric conditions Mode: UV Analytical wavelength: 247 nm Cell: 1 cm Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C15H20N2O4S in the Tablets taken: Result = (AU/AS) (CS/CU) (100/L) AU = absorbance of the Sample solution = absorbance of the Standard solution AS = concentration of the Standard solution (g/mL) CS CU = concentration of the Sample solution (tablet/mL) L = label claim (mg/tablet) Acceptance criteria: 93.0%107.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: pH 7.6 phosphate buffer (see pH 791); 900 mL Apparatus 1: 100 rpm Time: 60 min Detector: UV 245 nm Standard solution: USP Acetohexamide RS in Medium Sample solution: Sample per Dissolution 711; dilute with Medium to a concentration that is similar to that of the Standard solution. Analysis: Determine the amount of C15H20N2O4S dissolved, by using Medium as the blank, in comparison with the Standard solution. Tolerances: NLT 75% (Q) of the labeled amount of C15H20N2O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Acetohexamide RS
Acetohexamide Tablets
(Comment on this Monograph)id=m518=Acetohexamide Tablets=A-Monos.pdf) DEFINITION Acetohexamide Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of C15H20N2O4S. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample preparation: Evaporate 20.0 mL of Sample stock solution 2 in the Assay to dryness, use the residue in the procedure. ASSAY PROCEDURE Standard solution: 10 g/mL of USP Acetohexamide RS Sample stock solution 1: Weigh and finely powder NLT 20 Tablets. Transfer a portion of powder equivalent to 500 mg of acetohexamide to a 100-mL volumetric flask, add 60 mL of 0.1 N sodium hydroxide, shake for 30 min, dilute with water to volume and filter, discarding the first 20 mL of the filtrate (5 mg/mL). Sample stock solution 2: Transfer 20.0 mL of Sample stock solution 1 to a separator, add 2 mL of 3 N hydrochloric acid, extract with four 40-mL portions of chloroform, filtering each portion through chloroform-washed paper into a 200mL volumetric flask. Dilute with chloroform to volume (0.5 mg/mL). Sample stock solution 3: Evaporate 20.0 mL of Sample stock solution 2 to dryness, transfer the residue, with the aid of 0.1 N sodium hydroxide, to a 100-mL volumetric flask, and dilute with 0.1 N sodium hydroxide to volume (0.1 mg/mL).
Acetohydroxamic Acid
(Comment on this Monograph)id=m540=Acetohydroxamic Acid=A-Monos.pdf)
C2H5NO2 N-Acetyl hydroxyacetamide; Acetohydroxamic acid [546-88-3].
75.07
DEFINITION Acetohydroxamic Acid, dried over phosphorus pentoxide for 16 h, contains NLT 98.0% and NMT 101.0% of C2H5NO2. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solution: 20 mg/mL in water Analysis: To 10 mL of Sample solution, add 2 drops of potassium permanganate TS. Acceptance criteria: The pink color of the permanganate disappears.
46
Acetohydroxamic / Official Monographs
USP 32
the best-fit straight line from the fluorescence intensities of the three Standard solutions versus the hydroxylamine hydrochloride concentrations, in g/mL. From the best-fit straight line, determine the concentration, in g/mL, of hydroxylamine hydrochloride in the Sample solution. Calculate the percentage of hydroxylamine in the portion of Acetohydroxamic Acid taken: Result = (CU/C) (Mr1/Mr2) 100 = measured concentration of hydroxylamine hydrochloride in the Sample solution (g/mL) C = concentration of Acetohydroxamic Acid in the Sample solution (g/mL) = molecular weight of hydroxylamine, 33.03 Mr1 = molecular weight of hydroxylamine Mr2 hydrochloride, 69.50 Acceptance criteria: NMT 0.5% CU SPECIFIC TESTS LOSS ON DRYING 731: Dry it over phosphorus pentoxide for 16 h: it loses NMT 1.0% of its weight. COMPLETENESS OF SOLUTION 641: A 1.0-g portion dissolves in 10 mL of water to yield a clear solution COLOR OF SOLUTION Sample solution: 200 mg/mL Blank: Water Spectrometric conditions Mode: Spectroscopy Cell: 1 cm Analytical wavelength: Between 400 and 750 nm Analysis Sample: Sample solution Measure the absorbance. Acceptance criteria: NMT 0.050 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a cool, dry place. USP REFERENCE STANDARDS 11 USP Acetohydroxamic Acid RS
ASSAY PROCEDURE Solution A: 20 mg/mL of ferric chloride in 0.1 N hydrochloric acid Standard solution: 500 g/mL of USP Acetohydroxamic Acid RS in 0.1 N hydrochloric acid Sample solution: 500 g/mL of Acetohydroxamic Acid in 0.1 N hydrochloric acid Blank: 0.1 N hydrochloric acid Analysis: Transfer 10.0 mL each of the Standard solution, the Sample solution, and Blank, to separate 100-mL volumetric flasks. To each flask add 50 mL of 0.1 N hydrochloric acid and 10.0 mL of Solution A, and dilute with 0.1 N hydrochloric acid to volume. [NOTEWithout delay, concomitantly determine the absorbances.] Spectrometric conditions Mode: UV Analytical wavelength: 502 nm Analysis Samples: Standard solution and Sample solution Calculate the percentage of C2H5NO2 in the portion of Acetohydroxamic Acid taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Acetohydroxamic Acid RS in the Standard solution (g/mL) = concentration of Acetohydroxamic Acid in the CU Sample solution (g/mL) Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method I 231: NMT 20 ppm Sample solution: 40 mg/mL in a mixture of 1 N acetic acid and water (2:23) Organic Impurities PROCEDURE: LIMIT OF HYDROXYLAMINE Solution A: Dissolve 1.36 g of monobasic potassium phosphate in 950 mL of water, adjust with 1 M potassium hydroxide to a pH of 7.4, and dilute with water to 1000 mL. Solution B: 1 mg/mL of pyridoxal 5-phosphate monohydrate in Solution A in a low-actinic flask [NOTE Prepare fresh before use.] Standard stock solution: 2.0 mg/mL of hydroxylamine hydrochloride Standard solutions: Transfer 5.0, 10.0, and 15.0 mL of the Standard stock solution, to separate 100-mL volumetric flasks, and dilute with water to volume. Sample solution: Transfer 1500 mg of Acetohydroxamic Acid, previously dried, to a 100-mL beaker, and dissolve in a sufficient amount of water to cover the electrode of a calibrated pH meter (60 mL). While stirring, adjust with 0.05 M potassium hydroxide to a pH of 7.4. Transfer the contents of the beaker, with the aid of small portions of water, to a 100-mL volumetric flask, and dilute with water to volume. Blank: Water Analysis: Transfer 2.0 mL of each Standard solution, the Sample solution, and Blank into separate 100-mL volumetric flasks. To each flask, add 4.0 mL of Solution B. After 8 min, accurately timed, dilute the contents of each flask with Solution A to volume. Immediately determine the fluorescence intensities of the solutions from the Standard solutions and the Sample solution in a fluorometer at an excitation wavelength of 350 nm and an emission wavelength of 450 nm, setting the instrument to zero with the reagent blank. Determine AU AS CS
Acetohydroxamic Acid Tablets

(Comment on this Monograph)id=m548=Acetohydroxamic Acid Tablets=A-Monos.pdf) DEFINITION Acetohydroxamic Acid Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C2H5NO2. IDENTIFICATION Tablets produce a purple color when mixed with an acidic solution of ferric chloride. ASSAY PROCEDURE Solution A: 20 mg/mL of ferric chloride in 0.1 N hydrochloric acid Standard solution: 500 g/mL of USP Acetohydroxamic Acid RS in 0.1 N hydrochloric acid Sample solution: Transfer an equivalent to 500 mg of acetohydroxamic acid, from finely powdered Tablets (NLT 20), to a 1000-mL volumetric flask, add 500 mL of 0.1 N hydrochloric acid, and shake for 1 min. Dilute with 0.1 N hydrochloric acid to volume, and mix. Filter, discarding the first 40 mL of the filtrate. Use the clear filtrate. Blank: 0.1 N hydrochloric acid Analysis: Transfer 10.0 mL each of the Standard solution, the Sample solution, and Blank, to separate 100-mL volumetric flasks. To each flask add 50 mL of 0.1 N
USP 32
hydrochloric acid and 10.0 mL of Solution A. Dilute with 0.1 N hydrochloric acid to volume. [NOTEWithout delay, concomitantly determine the absorbances.] Spectrometric conditions Mode: UV Analytical wavelength: 502 nm Analysis Samples: Standard solution and Sample solution Calculate the percentage of C2H5NO2 in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Acetohydroxamic Acid RS in the Standard solution (g/mL) = nominal concentration of acetohydroxamic acid CU in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% AU AS CS PERFORMANCE TESTS DISSOLUTION 711, Procedure for a Pooled Sample Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Standard solution: USP Acetohydroxamic Acid RS in the same Medium Analysis: Determine the amount of C2H5NO2 dissolved, as directed in the Analysis under the Assay, using a filtered portion of the Sample solution in comparison with a Standard solution having a known concentration of USP Acetohydroxamic Acid RS. Tolerances: NLT 85% (Q) of the labeled amount of C2H5NO2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF HYDROXYLAMINE Solution A: Dissolve 1.36 g of monobasic potassium phosphate in 950 mL of water, adjust with 1 M potassium hydroxide to a pH of 7.4, and dilute with water to 1000 mL. Solution B: 1 mg/mL of pyridoxal 5-phosphate monohydrate in Solution A in a low-actinic flask [NOTE Prepare fresh before use.] Standard stock solution: 2.0 mg/mL of hydroxylamine hydrochloride in water Standard solutions: Transfer 5.0, 10.0, and 15.0 mL of the Standard stock solution, to separate 100-mL volumetric flasks, and dilute with water to volume. Sample solution: Transfer an equivalent to 1500 mg of acetohydroxamic acid, from finely powdered Tablets (NLT 20), to a 50-mL stoppered centrifuge tube. Add 30.0 mL of water, shake for 2 min, and centrifuge. Pipet 15.0 mL of the clear solution into a 50-mL beaker, add just enough water to cover the electrode of a calibrated pH meter, and while stirring, adjust with 0.5 M potassium hydroxide to a pH of 7.4. Transfer the contents of the beaker, with the aid of small portions of water, to a 50-mL volumetric flask, and dilute with water to volume. Blank: Water Analysis: Transfer 2.0 mL of each Standard solution, the Sample solution, and Blank into separate 100-mL volumetric flasks. To each flask, add 4.0 mL of Solution B. After 8 min, accurately timed, dilute the contents of each flask with Solution A to volume. Immediately determine the fluorescence intensities of the solutions from the Standard solutions and the Sample solution in a fluorometer at an excitation wavelength of
Official Monographs / Acetylcholine 47

350 nm and an emission wavelength of 450 nm, setting the instrument to zero with the reagent blank. Determine the best-fit straight line from the fluorescence intensities of the three Standard solutions versus the hydroxylamine hydrochloride concentrations, in g/mL. From the best-fit straight line, determine the concentration, in g/mL, of hydroxylamine hydrochloride in the Sample solution. Calculate the percentage of H3NO in the portion of Tablets taken: Result = (CS/CU) (Mr1/Mr2) 100 = concentration of USP Hydroxylamine Hydrochloride RS in the Standard solution (g/mL) CU = nominal concentration of hydroxylamine hydrochloride in the Sample solution (g/mL) Mr1 = molecular weight of hydroxylamine, 33.03 = molecular weight of hydroxylamine Mr2 hydrochloride, 69.50 Acceptance criteria: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Acetohydroxamic Acid RS CS
Acetylcholine Chloride
(Comment on this Monograph)id=m680=Acetylcholine Chloride=A-Monos.pdf)
C7H16ClNO2 181.66 Ethanaminium, 2-(acetyloxy)-N, N, N-trimethyl-, chloride; Choline acetate (ester) chloride. [60-31-1]. DEFINITION Acetylcholine Chloride contains NLT 98.0% and NMT 102.0% of C7H16ClNO2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solution: 100 mg/mL in water Analysis: To 5 mL of Sample solution, add 5 mL of silver nitrate TS: a white, curdy precipitate, which is soluble in ammonium hydroxide but insoluble in nitric acid, is formed. ASSAY PROCEDURE Sample: 400 mg of Acetylcholine Chloride Analysis: Dissolve in 15 mL of water in a glass-stoppered conical flask, add 40.0 mL of 0.1 N sodium hydroxide VS, and heat on a steam bath for 30 min. Insert the stopper, allow to cool, add phenolphthalein TS, and titrate the excess alkali with 0.1 N sulfuric acid VS. Perform a blank determination (see Residual Titrations under Titrimetry 541). Each mL of 0.1 N sodium hydroxide is equivalent to 18.17 mg of C7H16ClNO2. Acceptance criteria: 98.0%102.0% OTHER COMPONENTS CONTENT OF CHLORIDE: Transfer 280 mg to a porcelain casserole, and add 140 mL of water and 1 mL of
48
Acetylcholine / Official Monographs

dichlorofluorescein TS. Titrate with 0.1 N silver nitrate VS until the silver chloride flocculates and the mixture acquires a faint pink color. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Cl. NLT 19.3% and NMT 19.8% of Cl, calculated on the dried basis.
USP 32
ASSAY PROCEDURE Mobile phase: Add 1.03 g of sodium 1-heptanesulfonate to a mixture of 900 mL of water and 10 mL of methanol. Mix, then add sufficient glacial acetic acid and ammonium hydroxide, if necessary, to adjust the solution to a pH of 4.0. Add 50 mL of acetonitrile, then add water to make 1000 mL. [NOTESlight variation of the amount of acetonitrile may be required to improve resolution or adjust retention time.] System suitability solution: 0.2% solution of each of acetylcholine chloride and choline chloride Standard solution: A quantity of USP Acetylcholine Chloride RS in Mobile phase, to obtain a solution having a known concentration equal to that of the acetylcholine chloride in the Sample solution Sample solution: Transfer the contents of 1 container of Acetylcholine Chloride for Ophthalmic Solution to a 10-mL volumetric flask with the aid of Mobile phase, and add Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: 3.9 mm 30 cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 2.0 between acetylcholine chloride and choline chloride, System suitability solution Relative standard deviation: NMT 3.5%, Standard solution Analysis Samples: Standard solution and Sample Solution Calculate the percentage of C7H16ClNO2 in the container taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Acetylcholine Chloride RS in the Standard solution (mg/mL) CU = nominal concentration of acetylcholine chloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements rU rS CS
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
NMT 0.2%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 149152 ACIDITY: Dissolve 100 mg in 10 mL of recently boiled water, and add at once 1 drop of bromothymol blue TS: NMT 0.50 mL of 0.010 N sodium hydroxide is required in order to produce a color change. LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in a tight container, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetylcholine Chloride RS
Acetylcholine Chloride for Ophthalmic Solution

(Comment on this Monograph)id=m700=Acetylcholine Chloride for Ophthalmic Solution=A-Monos.pdf) DEFINITION Acetylcholine Chloride for Ophthalmic Solution is a sterile mixture of Acetylcholine Chloride with Mannitol or other suitable diluent, prepared by freeze-drying. Each container contains NLT 90.0% and NMT 115.0% of the labeled amount of acetylcholine chloride (C7H16ClNO2). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of aluminum oxide Standard solution: 10 mg/mL of USP Acetylcholine Chloride RS Sample solution: 10 mg/mL of acetylcholine chloride Spray reagent A: 5 mg/mL of cobaltous chloride in a mixture of alcohol and water (1:1) [NOTEThis solution is freshly prepared.] Spray reagent B: 10 mg/mL of potassium ferrocyanide in a mixture of alcohol and water (1:1) Application volume: 2 L Developing solvent system: Upper layer obtained by mixing water, butyl alcohol, and glacial acetic acid (5:4:1) [NOTEAllow to separate completely.] Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram, without delay, in a vapor-saturated chamber. Allow the solvent front to move about 10 cm beyond the initial spotting line. Immediately spray the plate with Spray reagent A. Dry the plate as before, and immediately spray the plate with Spray reagent B. Acceptance criteria: The RF value and color of the principal spot obtained from the Sample solution correspond to those obtained from the Standard solution. B. PROCEDURE Sample solution: 10 mg/mL of acetylcholine chloride Analysis: To 2 mL of Sample solution, add 1 drop of nitric acid and 1 mL of silver nitrate TS. Acceptance criteria: A curdy, white precipitate, soluble in an excess of 6 N ammonium hydroxide, is formed.
SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements. ACIDITY: Equivalent to 100 mg of acetylcholine chloride in 10 mL of recently boiled water Add at once 1 drop of bromothymol blue TS: NMT 0.50 mL of 0.010 N sodium hydroxide is required in order to produce a color change. WATER DETERMINATION, Method I 921: Perform the titration in the original container, observing precautions against contact with water or moist atmosphere. Adjust the concentration of the reagent so that the titration volume approaches but does not exceed the capacity of the container. Titrate to an amber color that persists for 15 s after mixing. Acceptance criteria: NMT 1.0% CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions.
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers as described under Injections 1, Containers for Sterile Solids, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetylcholine Chloride RS RU RS CS
Official Monographs / Acetylcysteine 49

= internal standard ratio of the Sample solution = internal standard ratio of the Standard solution = concentration of USP Acetylcysteine RS in the Standard solution (mg/mL) CU = concentration of Acetylcysteine in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Sample: 2 g Analysis: Transfer the Sample to a tared fused silica dish, heat on a hot plate until thoroughly charred, cool, add 1 mL of sulfuric acid, and heat gently until fuming ceases. Ignite at 600 until the carbon is consumed. Acceptance criteria: NMT 0.5% HEAVY METALS, Method II 231: NMT 10 ppm [CAUTIONExercise care because explosion may occur.] [NOTEIn a dropwise manner, wet the Sample with 2 mL of nitric acid, and proceed as directed for the Sample solution.] SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +21 to +27 pH 7.0 Buffer solution: Mix 29.5 mL of 1 N sodium hydroxide, 50 mL of 1 M monobasic potassium phosphate, and sufficient water to make 100 mL, and using a pH meter, adjust to a pH of 7.0 0.1 by adding more of either solution as necessary. Sample solution: In a 25-mL volumetric flask, mix 1.25 g with 1 mL of edetate disodium solution (1 in 100), add 7.5 mL of sodium hydroxide solution (1 in 25), and mix to dissolve. Dilute with pH 7.0 Buffer solution to volume. PH 791: 2.02.8, in a solution (1 in 100) LOSS ON DRYING 731: Dry it at a pressure of about 50 mm of mercury at 70 for 4 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetylcysteine RS USP L-Phenylalanine RS
Acetylcysteine
(Comment on this Monograph)id=m750=Acetylcysteine=AMonos.pdf)
L-Cysteine, N-acetyl-; N-Acetyl-L-cysteine [616-91-1].
C5H9NO3S
163.20
DEFINITION Acetylcysteine contains NLT 98.0% and NMT 102.0% of C5H9NO3S, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Mobile phase: 6.8 mg/mL of monobasic potassium phosphate Pass through a membrane filter having a 0.45-m porosity, and degas. Adjust with phosphoric acid to a pH of 3.0. Solution A: 0.5 mg/mL of sodium metabisulfite Internal standard solution: 5 mg/mL of USP LPhenylalanine RS in freshly-prepared Solution A Standard stock solution: 10 mg/mL of USP Acetylcysteine RS in Solution A Standard solution: 0.5 mg/mL of USP Acetylcysteine RS Pipet 10.0 mL of Standard stock solution and 10.0 mL of Internal standard solution to a 200-mL volumetric flask, and dilute with Solution A to volume. Sample stock solution: 10 mg/mL of Acetylcysteine in Solution A Sample solution: 0.5 mg/mL of Acetylcysteine Pipet 10.0 mL of Sample stock solution and 10.0 mL of Internal standard solution to a 200-mL volumetric flask, and dilute with Solution A to volume. Chromatographic system Mode: LC Detector: UV 214 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 5 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetylcysteine and Lphenylalanine are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 6 between acetylcysteine and DLphenylalanine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C5H9NO3S in the portion of Acetylcysteine taken: Result = (RU/RS) (CS/CU) 100
Acetylcysteine Solution
(Comment on this Monograph)id=m780=Acetylcysteine Solution=A-Monos.pdf) DEFINITION Acetylcysteine Solution is a sterile solution of Acetylcysteine in water, prepared with the aid of Sodium Hydroxide. It contains NLT 90.0% and NMT 110.0% of the labeled amount of acetylcysteine (C5H9NO3S). IDENTIFICATION INFRARED ABSORPTION 197K Sample solution: Place 10 mL in a suitable beaker, and adjust to a pH of 2 (pH indicator paper) using 3 N hydrochloric acid. Add up to 2 g of finely powdered sodium chloride, in two portions of 200 mg each initially, and then in smaller portions of 25 mg, stirring after each addition until the sodium chloride dissolves and a precipitate is formed. [NOTEThe precipitate appears as a very fine powder, and the solution turns cloudy. If no precipitate forms, add an additional drop of 3 N hydrochloric acid, and stir until the precipitate forms.] Allow to stand at room temperature for 15 min, and collect the residue by suction filtration. Use the acetylcysteine so obtained, after being dried at a pressure of 50 mm of mercury at 70 for 4 h.
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Acetylcysteine / Official Monographs
USP 32
ASSAY PROCEDURE Mobile phase: 6.8 mg/mL monobasic potassium phosphate in water Adjust with phosphoric acid to a pH of 3.0 Solution A: 0.5 mg/mL of sodium metabisulfite solution in water Internal standard solution: 5 mg/mL of USP LPhenylalanine RS in Solution A Standard stock solution: 10 mg/mL of USP Acetylcysteine RS in Solution A Standard solution: 0.5 mg/mL of USP Acetylcysteine RS in Solution A, from Standard stock solution, Internal standard solution and Solution A (1:1:18) Sample stock solution: Equivalent to 10 mg/mL of Acetylcysteine, from volume of solution in Solution A Sample solution: 0.5 mg/mL of acetylcysteine in Solution A, from Sample stock solution, Internal standard solution, and Solution A (1:1:18) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 6 between acetylcysteine and DLphenylalanine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution [NOTEThe relative retention times for acetylcysteine and L-phenylalanine are about 0.5 and 1.0, respectively.] Calculate the percentage of C5H9NO3S in each mL of the solution taken: Result = (RU/RS) (CS/CU) 100 = ratio of the peak response of acetylcysteine to that of L-phenylalanine of the Sample solution = ratio of the peak response of acetylcysteine to RS that of L-phenylalanine of the Standard solution = concentration of USP Acetylcysteine RS in the CS Standard solution (mg/mL) = nominal concentration of acetylcysteine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS PH 791: 6.07.5 STERILITY 71: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-unit or multipleunit tight containers that effectively exclude oxygen, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acetylcysteine RS USP L-Phenylalanine RS
Acetylcysteine and Isoproterenol Hydrochloride Inhalation Solution

(Comment on this Monograph)id=m810=Acetylcysteine and Isoproterenol Hydrochloride Inhalation Solution=A-Monos.pdf) DEFINITION Acetylcysteine and Isoproterenol Hydrochloride Inhalation Solution is a sterile solution of Acetylcysteine and Isoproterenol Hydrochloride in water. It contains NLT 90.0% and NMT 110.0% of the labeled amount of acetylcysteine (C5H9NO3S), and NLT 90.0% and NMT 115.0% of the labeled amount of isoproterenol hydrochloride (C11H17NO3 HCl). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample solution: Place 2 mL in a 10-mL beaker, and adjust with 3 N hydrochloric acid to a pH of 3 (pH indicator paper). Add 500 mg to 1 g of finely powdered sodium chloride, in two portions of 200 mg each initially, and then in smaller portions of 25 mg, stirring after each addition until the sodium chloride dissolves and a precipitate is formed. Allow to stand at room temperature for 15 min, and collect the residue by suction filtration. Use the acetylcysteine so obtained, after being dried at a pressure of 50 mm of mercury at 70 for 4 h. B. PROCEDURE Ferro-citrate solution and Buffer solution: Prepare as directed under Epinephrine Assay 391. Sample solution: Equivalent to 0.26 mg of isoproterenol hydrochloride from a volume of Inhalation Solution, in a test tube with 3 mL of 0.1 M mercuric chloride Analysis: Add 100 L of Ferro-citrate solution and 1.0 mL of Buffer solution Acceptance criteria: The presence of isoproterenol hydrochloride is confirmed by the development of a purple color. ASSAY ACETYLCYSTEINE Mobile phase: 6.8 mg/mL of monobasic potassium phosphate in water Adjust with phosphoric acid to a pH of 3.0 Solution A: 0.5 mg/mL of sodium metabisulfite solution in water Internal standard solution: 5 mg/mL of USP LPhenylalanine RS in Solution A Standard stock solution: 10 mg/mL of USP Acetylcysteine RS in Solution A Standard solution: 0.5 mg/mL of USP Acetylcysteine RS in Solution A, from Standard stock solution, Internal standard solution, and Solution A (1:1:18) Sample stock solution: Equivalent to 10 mg/mL of acetylcysteine, from volume of Inhalation Solution in Solution A Sample solution: 0.5 mg/mL of acetylcysteine in Solution A, from Sample stock solution, Internal standard solution, and Solution A (1:1:18) Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 214 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 5 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetylcysteine and L-phenylalanine are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 6 between acetylcysteine and DLphenylalanine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C5H9NO3S in each mL of the Inhalation Solution taken: Result = (RU/RS) (CS/CU) 100 = ratio of the peak response of acetylcysteine to that of L-phenylalanine from the Sample solution = ratio of the peak response of acetylcysteine to RS that of L-phenylalanine from the Standard solution = concentration of USP Acetylcysteine RS in the CS Standard solution (mg/mL) = nominal concentration of acetylcysteine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% ISOPROTERENOL HYDROCHLORIDE Solution A: 13.6 mg/mL of monobasic potassium phosphate in water Mobile phase: Methanol and Solution A (1:49) Internal standard solution: 150 mg of acetaminophen in a 500-mL volumetric flask. Add 5 mL of glacial acetic acid, and dilute with water to volume. Standard stock solution: 0.15 mg/mL of USP Isoproterenol Hydrochloride RS, in 0.05 M sodium metabisulfite Standard solution: 0.6 mg/mL of USP Isoproterenol Hydrochloride RS, from Standard stock solution, Internal standard solution, and 0.2 M acetic acid (2:2:1) Sample solution: Equivalent to 1.5 mg of isoproterenol hydrochloride from a volume of Inhalation Solution, and 10 mL of Internal standard solution to a 25-mL volumetric flask. Add dilute glacial acetic acid (1 in 100) to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 40-cm; packing L1 Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for isoproterenol hydrochloride and acetaminophen are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 6 between isoproterenol hydrochloride and acetaminophen Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H17NO3 HCl in each mL of the Inhalation Solution taken: Result = (RU/RS) (CS/CU) 100 RU RU
Official Monographs / Acitretin 51

= ratio of the peak responses of isoproterenol hydrochloride to acetaminophen from the Sample solution = ratio of the peak responses of USP Isoproterenol RS Hydrochloride RS to acetaminophen from the Standard solution = concentration of USP Isoproterenol CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of isoproterenol CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS PH 791: 6.07.0 STERILITY TESTS 71: It meets the requirements. COLOR AND CLARITY Standard solution: 0.100 N iodine VS and water (1:249) Sample solution: A portion of the Inhalation Solution Analysis: Visually analyze the Sample solution in a suitable clear glass test tube against a white background: it is not pinkish and it contains no precipitate. If any yellow color is observed in the Sample solution, concomitantly determine the absorbances of the Sample solution and the Standard solution in 1-cm cells with a suitable spectrophotometer set at 460 nm. Acceptance criteria: The absorbance of the Sample solution does not exceed that of the Standard solution. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, tightly closed with a glass or polyethylene closure, and store at controlled room temperature. LABELING: The label indicates that the Inhalation Solution is not to be used if its color is pinkish or darker than slightly yellow or if it contains a precipitate. USP REFERENCE STANDARDS 11 USP Acetylcysteine RS USP Isoproterenol Hydrochloride RS USP L-Phenylalanine RS
Acitretin
(Comment on this Monograph)id=m850=Acitretin=AMonos.pdf)
C21H26O3 2,4,6,8-Nonatetraenoic acid, 9-(4-methoxy-2,3,6trimethylphenyl)-3,7-dimethyl-, (all-E)-; (all-E)-9-(4-Methoxy-2,3,6-trimethylphenyl)-3,7dimethyl-2,4,6,8-nonatetraenoic acid [55079-83-9].
326.43
DEFINITION Acitretin contains NLT 98.0% and NMT 102.0% of C21H26O3, calculated on the dried basis. [CAUTIONAcitretin is a teratogen. Great care should be taken when handling to avoid inhalation of dust or contact with skin.] [NOTEUse low actinic glassware and perform all tests under yellow and subdued light.]
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Acitretin / Official Monographs
USP 32
Dilute quantitatively with alcohol. [NOTEStore the solution at 4 prior to injection.] Sample solution: 0.25 mg/mL of Acitretin in tetrahydrofuran, and alcohol (1:19) [NOTEStore the solution at 4 prior to injection.] System suitability (See Chromatography 621.) Sample: Standard solution [NOTEThe relative retention times for acitretin related compound A, acitretin, and acitretin related rompound B are 0.78, 1.0, and 1.61, respectively.] Suitability requirements Resolution: NLT 1.5 between acitretin related compound A and acitretin; NLT 1.5 between acitretin related rompound B and acitretin Relative standard deviation: NMT 10.0% for acitretin related compound A and NMT 10.0% for acitretin related compound B Analysis Samples: Standard solution and Sample solution Calculate the percentage of acitretin related compound A and acitretin related compound B in the portion of Acitretin taken: Result = (rU/rS) (CS/CU) 100 = peak response of the relevant impurity in the chromatogram of the Sample solution = peak response of the relevant impurity in the rS chromatogram of the Standard solution = concentration of USP Acitretin Related CS Compound A RS or USP Acitretin Related Compound B RS in the Standard solution (g/mL) = concentration of Acitretin in the Sample solution CU (g/mL) Calculate the percentage of impurities other than acitretin related compounds A and B in the portion of Acitretin taken Result = (rU/rS) (CS/CU) 100 = peak response of each individual unspecified impurity in the Sample solution = peak response of USP Acitretin RS in the rS Standard solution = concentration of USP Acitretin RS in the CS Standard solution (g/mL) CU = concentration of Acitretin in the Sample solution (g/mL) Acceptance criteria: NMT 0.3% of acitretin related compound A, NMT 0.3% of acitretin related compound B; NMT 0.1% of any individual unspecified impurity, and NMT 0.4% of total unspecified impurities Total impurities: NMT 1.0% rU SPECIFIC TESTS LOSS ON DRYING 731: Dry it in vacuum at a pressure not exceeding 19 mm of mercury at 100 for 4 h: it loses NMT 0.2% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. Store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Acitretin RS USP Acitretin Related Compound A RS USP Acitretin Related Compound B RS rU
IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Alcohol, glacial acetic acid, and water (92:0.3:8) System suitability solution: In a 200-mL volumetric flask, dissolve 2.0 mg each of USP Acitretin RS and USP Tretinoin RS in tetrahydrofuran. Dilute with alcohol to volume. Pipet 5.0 mL of this solution into a 200-mL volumetric flask, and dilute quantitatively with alcohol. [NOTEStore the solution at 4 prior to injection.] Standard solution: 0.1 mg/mL of USP Acitretin RS in alcohol [NOTEUse tetrahydrofuran to dissolve USP Acitretin RS before diluting with alcohol. The final concentration of tetrahydrofuran in the preparation will be 2%. Store the solution at 4 prior to injection.] Sample stock solution: 0.25 mg/mL of Acitretin in tetrahydrofuran and alcohol (1:19) Sample solution: 0.1 mg/mL of Acitretin from Sample stock solution in alcohol [NOTEStore the solution at 4 prior to injection.] Chromatographic system Mode: LC Detector: UV 360 nm Column: 4-mm 25-cm; packing L1 Flow rate: 0.6 mL/min Injection size: 10 L System suitability (See Chromatography 621, System Suitability.) Sample: System suitability solution [NOTEThe relative retention times for tretinoin and for acitretin are 0.84 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 tretinoin and acitretin in System suitability solution Relative standard deviation: NMT 1.0% Acitretin Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H26O3 in the portion of Acitretin taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acitretin RS in the Standard solution (mg/mL) = concentration of Acitretin in the Sample solution CU (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method I 231: NMT 20 ppm Organic Impurities PROCEDURE Mobile phase and Chromatographic system: Proceed as directed in the Assay. Standard solution: 0.8 g/mL each of USP Acitretin RS, USP Acitretin Related Compound A RS, and USP Acitretin Related Compound B RS in tetrahydrofuran
USP 32
USP Tretinoin RS
Official Monographs / Acitretin 53

Mode: LC Detector: UV 365 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 25 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for acitretin related compound A, acitretin, and 9-cis isomer are 0.84, 1.0, and 1.09, respectively.] Suitability requirements Resolution: NLT 3.0 between acitretin related compound A and acitretin; NLT 1.8 between the 9-cis isomer and acitretin Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H26O3 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acitretin RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (Capsules/mL) Acceptance criteria: 90.0%110.0% rU rS CS IMPURITIES Organic Impurities LIMIT OF DEGRADATION PRODUCTS Diluent, Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Sample solution: Use the Sample solution, prepared as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each degradation product in the portion of Capsules taken: Result = (rU/rT) 100 = peak response for each individual impurity rU rT = sum of the responses of all of the peaks Acceptance criteria: NMT 0.5% of acitretin related compound A, NMT 0.4% of any individual unspecified impurity, and NMT 0.8% of total unspecified impurities is found. PERFORMANCE TESTS DISSOLUTION 711 Medium: 3% sodium lauryl sulfate in deaerated water, pH 9.6 to 10.0; 900 mL Apparatus 1: 100 rpm Time: 30 min Determine the amount of acitretin (C21H26O3) dissolved employing the following method. Sample solutions: Sample per 711 Dissolution. Dilute with Medium to a concentration that is similar to the Standard solution. Use portions of the solution under test passed through a suitable 0.45-m filter. Standard solution: 14 mg of USP Acitretin RS in alcohol, and Medium (1:9). For Capsules labeled to contain 10 mg, transfer 20.0 mL of this solution to a 50-mL volumetric flask, and dilute with Medium to volume. Capsule shell solution: 6 clean empty-shell Capsules in 900 mL of Medium
Acitretin Capsules
(Comment on this Monograph)id=m854=Acitretin Capsules=AMonos.pdf) DEFINITION Acitretin Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of acitretin (C21H26O3). [CAUTIONAcitretin is a teratogen. Great care should be taken when handling to avoid inhalation of dust or contact with skin.] [NOTEUse low-actinic glassware and perform all tests under yellow and subdued light. Make all injections within 1 h of Sample solution.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 10 mg/mL of USP Acitretin RS in tetrahydrofuran Sample solution: Equivalent to 20 mg of acitretin from Capsules Grind to a fine powder, then triturate for 30 s with 2 mL of tetrahydrofuran. Transfer the suspension to a 12-mL conical centrifuge tube, and centrifuge to obtain a clear supernatant. Application volume: 10 L Developing solvent system: Chloroform and methanol (4:1) Analysis Samples: Standard solution and Sample solution Procedure: Proceed as directed in the chapter, and then air-dry. Spray the plate with a saturated solution of antimony trichloride in chloroform (25 g in 100 mL) followed by concentrated sulfuric acid, and then locate the spots. ASSAY PROCEDURE Diluent: Methanol and tetrahydrofuran (13:10) Mobile phase: Methanol, alcohol, glacial acetic acid, and water (74:5:0.5:21) System suitability solution: Transfer 2 mL of the Standard solution to a clear 4-mL glass vial. After sealing the vial with a teflon-lined silicone septum and cap, place the vial on its side in a light chamber, expose it to 400 foot-candles of fluorescent light for 5 min, and then completely wrap the vial with aluminum foil. [NOTEExposure to the fluorescent light allows for the formation of two degradation products: acitretin related compound A and the 9-cis isomer [(E,E,Z,E)-9-(4methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8nonatetraenoic acid.] Standard solution: 0.1 mg/mL of USP Acitretin RS in water and Diluent (2:23) [NOTEBefore make-up to final volume sonicate for 5 min.] Sample solution: Carefully separate and place both halves of 10 Capsules into a 100-mL volumetric flask. Stopper and shake the flask to remove the fill. Add 8 mL of water while rinsing any fill from the neck of the flask. Place the flask in a water bath set at 45 for 10 min, shaking initially and at 5min intervals up to 10 min. Place the resulting suspension in an ultrasonic bath for 15 min. Dilute with Diluent to volume, and sonicate for 5 additional min. Cool to room temperature and, if necessary, dilute with Diluent to volume. Filter the suspension, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.)
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Acitretin / Official Monographs
USP 32
Dissolve both in a volume of 0.1 N sodium hydroxide of NMT 10% of the final solution volume, and then dilute the solution to volume with water. System suitability solution B: 0.7 g/mL of guanine Dissolve in a volume of 0.1 N sodium hydroxide of NMT 10% of the final solution volume, and then dilute to volume with water. Guanine standard stock solution: Transfer 8.75 mg of guanine to a 500-mL volumetric flask. Dissolve in 50 mL of 0.1 N sodium hydroxide, and dilute with water to volume. Guanine standard solution: 0.7 g/mL guanine in 0.01 N sodium hydroxide from Guanine standard stock solution Standard stock solution: Dissolve 25 mg of USP Acyclovir RS in 5 mL of 0.1 N sodium hydroxide in a 50-mL volumetric flask, and dilute with water to volume. Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.01 N sodium hydroxide from Standard stock solution Sample stock solution: Dissolve 100 mg of Acyclovir in 20 mL of 0.1 N sodium hydroxide in a 200-mL volumetric flask, and dilute with water to volume. Sample solution: 0.1 mg/mL Acyclovir in 0.01 N sodium hydroxide from Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 3 mL/min Injection size: 20 L System suitability Samples: System suitability solution A and System suitability solution B Suitability requirements Resolution: NLT 2.0 between acyclovir and guanine from System suitability solution A Tailing factor: NMT 2, System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections of acyclovir for System suitability solution A; NMT 2.0% for replicate injections of System suitability solution B Analysis Samples: Standard solution, Guanine standard solution, and Sample solution Calculate the percentage of guanine in the portion of Acyclovir taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Guanine standard solution = concentration of guanine in the Guanine standard solution (g/mL) CU = concentration of the Sample solution (g/mL) Acceptance criteria: NMT 0.7% of guanine Calculate the percentage of C8H11N5O3 in the portion of Acyclovir taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acyclovir RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) rU rS CS
Analysis Samples: Standard solution, Blank, Capsule shell solution, and Sample solution Determine the amount of C21H26O3 dissolved by employing UV absorption at the wavelength of maximum absorbance at about 347 nm, using 2-mm cells, in comparison with the appropriate Standard solution. Use the Medium as a blank. Determine the absorbance of the Capsule shell solution under the same conditions. Calculate the amount of C21H26O3 dissolved: Result = [(AU ACS)/AS] (CS/CU) 100 AU ACS = absorbance of the Sample solution = capsule shell correction, calculated as shown below = absorbance of the appropriate Standard solution AS = concentration of the appropriate Standard CS solution (mg/mL) = concentration of the Sample solution CU (Capsules/mL) The Capsule shell correction, ACS, is calculated as follows: ACS = ACSS/N = absorbance of the Capsule shell solution = number of Capsule shells used to prepare the Capsule shell solution Tolerances: NLT 85% (Q) of the labeled amount of acitretin (C21H26O3) is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Acitretin RS ACSS N
Acyclovir
(Comment on this Monograph)id=m890=Acyclovir=AMonos.pdf)
C8H11N5O3 6H-Purin-6-one, 2-amino-1,9-dihydro-9-[(2hydroxyethoxy)methyl]-; 9-[(2-Hydroxyethoxy)methyl]guanine [59277-89-3].
225.20
DEFINITION Acyclovir contains NLT 98.0% and NMT 101.0% of C8H11N5O3, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY LIMIT FOR GUANINE Mobile phase: Glacial acetic acid in water (1 in 1000) System suitability solution A: 0.1 mg/mL each of USP Acyclovir RS and guanine
USP 32
Official Monographs / Acyclovir 55

Analysis: Separately inject the Standard solution and the Sample solution into the chromatograph, record the chromatograms, and measure the peak responses. Calculate the percentage of C8H11N5O3 in the Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Acyclovir RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 93.0%107.0% IMPURITIES Organic Impurities PROCEDURE [NOTEMobile phase, Sample solution, and Chromatographic system: proceed as directed in the Assay.] Analysis: Inject the Sample solution into the chromatograph, record the chromatograms, and measure the peak responses. Calculate the percentage of each impurity in the portion of Capsules taken: Result = (rU/rT) 100 = peak response for each impurity rU = sum of the responses for all the peaks rT Acceptance criteria Guanine: NMT 2.0% Any individual impurity: NMT 0.5% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 45 min Detector: UV 254 nm Sample solutions: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Acyclovir RS in Medium Analysis: Determine the amount of C8H11N5O3 dissolved from UV absorption at the wavelength of maximum absorption on filtered portions of the solution under test. Tolerances: NLT 75% (Q) of the labeled amount of C8H11N5O3 is dissolved. UNIFORMITY OF DOSAGE UNITS 905 Acceptance criteria: Meet the requirements for Content Uniformity ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 15 and 25. Protect from light and moisture. USP REFERENCE STANDARDS 11 USP Acyclovir RS rU rS CS
IMPURITIES Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Sample solution: 10 mg/mL in dimethyl sulfoxide Standard solutions: 0.01, 0.05, 0.1, and 0.2 mg/mL of USP Acyclovir RS in dimethyl sulfoxide Eluant: Chloroform, methanol, and ammonium hydroxide (80:20:2) Visualization: 1 Application volume: 5 L Acceptance criteria: NMT 1% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 6.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at room temperature. Protect from light and moisture. USP REFERENCE STANDARDS 11 USP Acyclovir RS
Acyclovir Capsules
(Comment on this Monograph)id=m893=Acyclovir Capsules=AMonos.pdf) DEFINITION Acyclovir Capsules contain NLT 93.0% and NMT 107.0% of the labeled amount of acyclovir (C8H11N5O3). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.02 M acetic acid System suitability solution A: 0.1 mg/mL each of USP Acyclovir RS and guanine in 0.1 N sodium hydroxide System suitability solution B: 2.0 g/mL of guanine in 0.1 N sodium hydroxide Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N sodium hydroxide Sample solution: Transfer the contents of Capsules equivalent to 10 mg of acyclovir (NLT 10 Capsules) to a 100-mL volumetric flask, dissolve in 10 mL of 0.1 N sodium hydroxide, dilute to volume with water, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.2-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution A and System suitability solution B [NOTEThe relative retention times for System suitability solution A for guanine and acyclovir are about 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between guanine and acyclovir for System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections of acyclovir using System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections of System suitability solution B
Acyclovir for Injection

(Comment on this Monograph)id=m894=Acyclovir for Injection=A-Monos.pdf) DEFINITION Acyclovir for Injection contains NLT 90.0% and NMT 110.0% of the labeled amount of acyclovir (C8H11N5O3).
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Acyclovir / Official Monographs
USP 32
System suitability solution: 0.5 g/mL each of purine and USP Acyclovir RS in Solution A Acyclovir standard solution: 5 g/mL of USP Acyclovir RS in Solution A Guanine solution: Dissolve 50 mg of guanine in 50 mL of 0.1 N sodium hydroxide in a 500-mL volumetric flask, and bring the solution to volume with water. Standard solution A: 0.5 g/mL of Acyclovir standard solution in Solution A Standard solution B: 5 g/mL of Guanine solution in Solution A Sample solution: 0.5 mg/mL of acyclovir in Solution A, prepared by constituting NLT 10 vials of Acyclovir for Injection and diluting in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: System suitability solution, Standard solution A, and Standard solution B [NOTETypical retention times for guanine and acyclovir of Standard solution A and Standard solution B are 5.8 min and 14 min, respectively.] Suitability requirements Resolution: NLT 2.0 between purine and acyclovir, System suitability solution Relative standard deviation: NMT 1% for the acyclovir peak area and the guanine peak area using replicate injections of Standard solution A and Standard solution B Analysis: Calculate the percentage of guanine in the Acyclovir for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response for guanine, if present, in the Sample solution = peak response of guanine in the Standard rS solution = concentration of guanine in the Standard CS solution (mg/mL) CU = concentration of acyclovir in the Sample solution (mg/mL, based on the label claim) Acceptance criteria: NMT 1.0% guanine Calculate the percentage of each other impurity in the Acyclovir for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response for each impurity = peak response of acyclovir in the Standard solution = concentration of USP Acyclovir RS in the CS Standard solution (mg/mL) = concentration of acyclovir in the Sample solution CU (mg/mL, based on the label claim) Acceptance criteria: NMT 0.15% for any peak having a relative retention time of about 0.7 compared to the acyclovir peak; NMT 0.5% for any other individual impurity; and NMT 1.0% for the total of all other impurities rU rS rU
IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.02 M acetic acid System suitability solution A: 0.1 mg/mL each of USP Acyclovir RS and guanine in 0.1 N sodium hydroxide System suitability solution B: 2.0 g/mL of guanine in 0.1 N sodium hydroxide Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N sodium hydroxide Sample solution: Constitute 1 vial of Acyclovir for Injection with water. Transfer an amount, equivalent to 10 mg of acyclovir, to a 100-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.2-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution A and System suitability solution B [NOTEThe relative retention times for guanine and acyclovir are 0.6 and 1.0, respectively, in the System suitability solution A.] Suitability requirements Resolution: NLT 2.0 between guanine and acyclovir, System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections for the acyclovir peak, System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections, System suitability solution B Analysis Calculate the percentage of C8H11N5O3 in the portion of Acyclovir for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Acyclovir RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 90.0%110.0% rU rS CS IMPURITIES Organic Impurities PROCEDURE Solution A: 0.17 M acetic acid and methanol (125:8) Make adjustments if necessary. Solution B: Methanol Mobile phase: Use variable mixtures of Solution A and Solution B as directed for Chromatographic system. Make adjustments to either solution as necessary.
Time (min) 0 15 45 46 56 Solution A (%) 100 100 65 100 100 Solution B (%) 0 0 35 0 0
USP 32
SPECIFIC TESTS PH 791: 11.012.5, 50 mg/mL of acyclovir WATER DETERMINATION Method I 921: NMT 5.5% STERILITY TESTS 71: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements BACTERIAL ENDOTOXINS TEST 85 NMT 0.174 USP Endotoxin Unit/mg of acyclovir OTHER REQUIREMENTS: Meets the requirements under Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 15 and 25. Protect from light. USP REFERENCE STANDARDS 11 USP Acyclovir RS USP Endotoxin RS rU rS CS
Official Monographs / Acyclovir 57

= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acyclovir RS in the Standard solution (mg/mL) CU = concentration of acyclovir in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF GUANINE [NOTEFor Mobile phase, Sample solution, and Chromatographic system, proceed as directed in the Assay.] Standard solution: 2.0 g/mL of guanine in 0.1 M sodium hydroxide Analysis Samples: Standard solution and Sample solution Calculate the percentage of guanine in the portion of Ointment taken: Result = (rU/rS) (CS/CU) 100 = peak response of guanine from the Sample solution = peak response of guanine from the Standard rS solution CS = concentration of guanine in the Standard solution (mg/mL) CU = concentration of acyclovir in the Sample solution (mg/mL) Acceptance criteria: NMT 2.0% rU SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED MICROORGANISMS 62 Acceptance criteria: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 15 and 25 in a dry place. USP REFERENCE STANDARDS 11 USP Acyclovir RS
Acyclovir Ointment
(Comment on this Monograph)id=m895=Acyclovir Ointment=A-Monos.pdf) DEFINITION Acyclovir Ointment contains NLT 90.0% and NMT 110.0% of the labeled amount of acyclovir (C8H11N5O3), in a suitable ointment base. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.02 M acetic acid System suitability solution A: 0.1 mg/mL each of USP Acyclovir RS and guanine in 0.1 N sodium hydroxide System suitability solution B: 2.0 g/mL of guanine in 0.1 N sodium hydroxide Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N sodium hydroxide Sample solution: Transfer an amount of Ointment, equivalent to 10 mg of acyclovir, to a 100-mL volumetric flask, dissolve in and dilute with 0.1 N sodium hydroxide to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 3 mL/min Injection size: 20 L System suitability Samples: System suitability solution A and System suitability solution B [NOTEThe relative retention times for guanine and acyclovir, are about 0.6 and 1.0, respectively, in System suitability solution A.] Suitability requirements Resolution: NLT 2.0 between guanine and acyclovir, System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections for the acyclovir peak, System suitability solution A; NMT 2.0% for replicate injections, System suitability solution B Analysis Samples: Standard solution and Sample solution Calculate the quantity, in percentage, of C8H11N5O3 in the portion of Ointment taken: Result = (rU/rS) (CS/CU) 100
Acyclovir Oral Suspension

(Comment on this Monograph)id=m898=Acyclovir Oral Suspension=A-Monos.pdf) DEFINITION Acyclovir Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of acyclovir (C8H11N5O3). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.02 M acetic acid System suitability solution A: 0.1 mg/mL each of USP Acyclovir RS and guanine in 0.1 N sodium hydroxide System suitability solution B: 2.0 g/mL of guanine in 0.1 N sodium hydroxide Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N sodium hydroxide
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Acyclovir / Official Monographs

Acceptance criteria: NMT 2.0%
USP 32
Sample stock solution: Transfer an amount of well-shaken Oral Suspension equivalent to 200 mg of acyclovir to a 200mL volumetric flask, add 100 mL of 0.1 N sodium hydroxide, shake by mechanical means for 15 min, and sonicate, if necessary, to dissolve the Oral Suspension completely. Dilute to volume with 0.1 N sodium hydroxide. Sample solution: Transfer 10.0 mL of the Sample stock solution to a 100-mL volumetric flask, and dilute to volume with water. Chromatographic system (See Chromatography 621, System Suitability). Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 3 mL/min Injection size: 20 L System suitability Sample: System suitability solution A and System suitability solution B [NOTEThe relative retention times for guanine for acyclovir are about 0.6 and 1.0, respectively, in System suitability solution A.] Suitability requirements Resolution: NLT 2.0 between guanine and acyclovir for System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections for the acyclovir peak using System suitability solution A Relative standard deviation: NMT 2.0% for replicate injections of System suitability solution B Analysis: Separately inject the Standard solution and the Sample solution into the chromatograph, record the chromatograms, and measure the peak responses. Calculate the percentage of C8H11N5O3 in the portion of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acyclovir RS in the Standard solution (mg/mL) = nominal concentration of acyclovir in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS IMPURITIES Organic Impurities PROCEDURE: LIMIT OF GUANINE [NOTEMobile phase, Sample solution, and Chromatographic system: proceed as directed in the Assay.] Standard solution: 2.0 g/mL of guanine in 0.1 M sodium hydroxide Analysis: Separately inject the Standard solution and the Sample solution into the chromatograph, record the chromatograms, and measure the peak responses. Calculate the percentage of guanine in the portion of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response for guanine from the Sample solution = peak response for guanine from the Standard solution = concentration of guanine in the Standard solution (mg/mL) = nominal concentration of acyclovir in the Sample solution (mg/mL)
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED MICROORGANISMS 62 Acceptance criteria: Its total count does not exceed 10 cfu/mL, and it meets the requirements of the tests for absence of Salmonella species and Escherichia coli. PH 791 Acceptance criteria: Between 4.5 and 7.0 PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 Acceptance criteria: Meets the requirements for Oral Suspension packaged in single-unit containers DELIVERABLE VOLUME 698 Acceptance criteria: Meets the requirements for Oral Suspension packaged in single-unit containers ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 15 and 25. Protect from light. USP REFERENCE STANDARDS 11 USP Acyclovir RS
Acyclovir Tablets
(Comment on this Monograph)id=m900=Acyclovir Tablets=AMonos.pdf) DEFINITION Acyclovir Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of acyclovir (C8H11N5O3). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.02 M acetic acid System suitability solution A: 0.1 mg/mL each of USP Acyclovir RS and guanine in 0.1 N sodium hydroxide System suitability solution B: 2.0 g/mL of guanine in 0.1 N sodium hydroxide Standard solution: 0.1 mg/mL of USP Acyclovir RS in 0.1 N sodium hydroxide Sample solution: Transfer an amount of finely powdered Tablets equivalent to 10 mg of acyclovir (NLT 10 Tablets) to a 100-mL volumetric flask, dissolve in 10 mL of 0.1 N sodium hydroxide, dilute with water to volume, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Column temperature: 40 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution A, and System suitability solution B [NOTEFor System suitability solution A, the relative retention times for guanine and acyclovir are about 0.6 and 1.0, respectively.]
USP 32
Suitability requirements Resolution: NLT 2.0 between guanine and acyclovir for System suitability solution A Relative standard deviation: NMT 2.0% for acyclovir peak for multiple injections of System suitability solution A; NMT 2.0% for multiple injections of System suitability solution B Analysis Samples: Standard solution and Sample solution Calculate the quantity, as a percentage, of C8H11N5O3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Acyclovir RS in the Standard solution (mg/mL) = nominal concentration of acyclovir in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 45 min Detector: UV 254 nm Standard solution: USP Acyclovir RS in Medium Sample solutions: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Analysis: Determine the amount of C8H11N5O3 dissolved from UV absorption at the wavelength on filtered portions of the solution under test. Tolerances: NLT 80% (Q) of the labeled amount of C8H11N5O3 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation IMPURITIES Organic Impurities PROCEDURE [NOTEFor Mobile phase, Standard solution, Sample solution, and Chromatographic system, proceed as directed in the Assay.] Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Tablets taken: Result = 100(rI/rS) = peak response for each impurity rI = sum of the responses for all of the peaks rS Acceptance criteria Individual impurities: NMT 2.0% for guanine, and NMT 0.5% for any other impurity ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 15 and 25. Protect from light and moisture. USP REFERENCE STANDARDS 11 USP Acyclovir RS
Official Monographs / Adenine 59
Adenine
(Comment on this Monograph)id=m930=Adenine=AMonos.pdf)
C5H5N5 1H-Purin-6-amine; 1,6-Dihydro-6-iminopurine [73-24-5].
135.13
DEFINITION Adenine contains NLT 98.0% and NMT 102.0% of C5H5N5, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE 0.1 N Perchloric acid: Standardize 0.1 N perchloric acid VS as follows: 300 mg of potassium biphthalate to a 150-mL beaker, and dissolve in 80 mL of a mixture of 100 mL of glacial acetic acid and 300 mL of acetic anhydride, by stirring. Titrate with the perchloric acid solution. Each 20.42 mg of potassium biphthalate is equivalent to 1 mL of 0.1 N perchloric acid. Sample: 200 mg of Adenine Analysis: Dissolve in 80 mL of a mixture of 100 mL of glacial acetic acid and 300 mL of acetic anhydride by stirring. Titrate with 0.1 N Perchloric acid. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N Perchloric acid is equivalent to 13.514 mg of C5H5N5. Acceptance criteria: 98.0%102.0% OTHER COMPONENTS NITROGEN CONTENT, Method II 461: calculated on the dried basis 50.2%53.4% is found,
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Solution A: Dissolve 4.54 g of monobasic potassium phosphate in water to make 500 mL of solution. Dissolve 4.73 g of anhydrous dibasic sodium phosphate in water to make 500 mL of solution. Mix 38.9 mL of the monobasic potassium phosphate solution with 61.1 mL of the dibasic sodium phosphate solution. Adjust, if necessary, by the dropwise addition of the dibasic sodium phosphate solution to a pH of 7.0. Standard stock solution: Dissolve a suitable quantity of USP Adenine RS in hot water, cool, and dilute quantitatively with water to obtain a solution having a known concentration of 0.19 mg/mL.
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Standard solutions: Pipet 5-mL portions into three 100mL volumetric flasks, dilute with 0.10 N hydrochloric acid, 0.010 N sodium hydroxide, and Solution A, respectively. Sample stock solution: Dissolve a suitable quantity of Adenine in hot water, cool, and dilute quantitatively with water to obtain a solution having a known concentration of 0.19 mg/mL. Sample solutions: Pipet 5-mL portions into three 100-mL volumetric flasks, dilute with 0.10 N hydrochloric acid, 0.010 N sodium hydroxide, and Solution A, respectively. Blank: Water Mode: Spectrometry Analytical wavelength: 220320 nm Cell: 1 cm Analysis Samples: Standard solutions and Sample solutions Acceptance criteria: The respective absorptivities, calculated on the dried basis, at the wavelengths of maximum absorbance, for each pair of corresponding solutions do not differ by more than 2.0%. Acceptance criteria: 99.0%101.0%
USP 32
SPECIFIC TESTS LOSS ON DRYING 731: 1.0% of its weight.
Dry it at 110 for 4 h: it loses NMT
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Adenine RS
Adenosine
(Comment on this Monograph)id=m938=Adenosine=AMonos.pdf)
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities Solution A: 6.8 g/L of potassium hydrogen sulfate and 3.4 g/L of tetrabutylammonium hydrogen sulfate in water. Adjust with 2 N potassium hydroxide to a pH of 6.5. Mobile phase: Solution A and (1 in 10,000) sodium azide solution (3:2) System suitability solution: 0.2 mg/mL each of Adenosine and inosine in Mobile phase Sample solution: 1.0 mg/mL of Adenosine in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution Suitability requirements Resolution: NLT 9.0 between adenosine and inosine, System suitability solution Tailing factor: NMT 2.5, System suitability solution Relative standard deviation: NMT 2.0%, System suitability solution [NOTEChromatograph the Sample solution, and adjust the run time to at least twice the retention time of the major peak.] Analysis Samples: Sample solution Determine the percentage of each impurity in the portion of Adenosine taken. Individual impurities: NMT 0.1% each of guanosine, inosine, and uridine, and NMT 0.2% of adenine Total impurities: NMT 0.5% SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 233238 OPTICAL ROTATION, Specific Rotation 781S: 68 to 72 Sample solution: 20 mg/mL in sodium hydroxide solution (1 in 20), determined on a Sample previously dried at 105 for 2 h ACIDITY OR ALKALINITY: Suspend 1000 mg in 20 mL of carbon dioxide-free water. Stir for 30 s, and pass through a coarse filter. To each of two 10-mL portions of the filtrate, add 0.1 mL of bromocresol purple TS. Acceptance criteria: NMT 0.3 mL of 0.01 N sodium hydroxide is required to produce a blue-violet color in one portion. NMT 0.1 mL of 0.01 N hydrochloric acid is required to produce a yellow color in the other portion. LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.5% of its weight. LIMIT OF AMMONIA Sample solution: Suspend 0.5 g in 10 mL of water. Stir for 30 s, and pass through a coarse filter. Dilute the filtrate with water to 15 mL, and use the filtrate.
C10H13N5O4 9--D-Ribofuranosyladenine; 6-Amino-9--D-ribofuranosyl-9-H-purine [58-61-7]. DEFINITION Adenosine contains NLT 99.0% and NMT 101.0% of C10H13N5O4, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197M
267.25
ASSAY PROCEDURE Sample: 200 mg of Adenosine previously dried at 105 for 2h Analysis: Dissolve in 50 mL of glacial acetic acid and titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 26.72 mg of C10H13N5O4.
USP 32
Standard stock solution: Dilute 1 mL of ammonium chloride solution (314 mg in 1000 mL) with 100 mL of water. Standard solution: Standard stock solution and water (2:13) Analysis: To the Sample solution and the Standard solution add 0.3 mL of alkaline mercuric-potassium iodide TS, cap the test tubes, and allow to stand for 5 min. Acceptance criteria: The Sample solution does not exhibit a more intense yellow color than that of the Standard solution (NMT 0.0004% ammonia). LIMIT OF CHLORIDE Sample solution: Suspend 0.2 g in 10 mL of water. Stir for 30 s, pass through a coarse filter, and use the filtrate. Standard solution: Dilute 1 mL of sodium chloride solution (231 mg in 1000 mL) with 100 mL of water. Analysis: To the Sample solution and 10 mL of the Standard solution, add 1 mL of nitric acid and 1 mL of silver nitrate TS, dilute each solution with water to 40 mL. Allow the solutions to stand for 5 min, protected from light. Acceptance criteria: When viewed against a dark background, the Sample solution is not more turbid than the Standard solution (NMT 0.007% chloride). LIMIT OF SULFATE Sample solution: Suspend 0.75 g in 15 mL of water. Stir for 30 s, pass through a coarse filter, and use the filtrate. Standard solution: Add 0.15 mL of 0.020 N sulfuric acid to 15 mL of water. Analysis: To the Sample solution and the Standard solution add 2 mL of barium chloride TS and 1 mL of 3 N hydrochloric acid, dilute each solution with water to 30 mL, and mix. Allow the solutions to stand for 5 min. Acceptance criteria: The Sample solution is not more turbid than the Standard solution (NMT 0.02% sulfate). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Adenosine RS
Official Monographs / Adenosine 61

anticipated final volume of the Standard solution before the addition of the warm water.] System sensitivity solution: Standard solution and water (3:197) Sample stock solution: Equivalent to 0.3 mg/mL of adenosine from volume of Injection, in water [NOTEReserve a portion of this stock solution for use in the test for Organic Impurities.] Sample solution: 0.03 mg/mL of adenosine, from Sample stock solution and water (1:9) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2.5 mL/min Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEChromatograph the System sensitivity solution and adjust the run time to 21/2 times the retention time of adenosine.] Suitability requirements Resolution: NLT 6.0 between adenosine and inosine, System suitability solution Tailing factor: NMT 2.0 for the adenosine peak, System suitability solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H13N5O4 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Adenosine RS in the Standard solution (mg/mL) = nominal concentration of adenosine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS IMPURITIES Organic Impurities PROCEDURE Mobile phase, System suitability solution, Standard solution, System sensitivity solution, and Chromatographic system: Proceed as directed in the Assay. Sample solution: Use the Sample stock solution reserved from the Assay. Analysis Sample: Sample solution Calculate the percentage of each impurity in the volume of Injection taken: Result = (ri/rs) 100 = peak response for each impurity ri = sum of the responses of all of the peaks rs Acceptance criteria Individual impurity: NMT 1.0% Total impurities: NMT 1.5% SPECIFIC TESTS PH 791: 4.57.5 PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections.
Adenosine Injection
(Comment on this Monograph)id=m940=Adenosine Injection=A-Monos.pdf) DEFINITION Adenosine Injection is a sterile solution of Adenosine in Water for Injection. It may contain Sodium Chloride. It contains NLT 90.0% and NMT 110.0% of the labeled amount of adenosine (C10H13N5O4). IDENTIFICATION The retention time of the adenosine peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 2.0 g of monobasic potassium phosphate in 800 mL of water. Add 5 mL of 1.0 M tetrabutylammonium dihydrogen phosphate solution, dilute with water to 980 mL, and mix. Add 20 mL of acetonitrile, mix, filter, and degas. Make adjustments if necessary. System suitability solution: 0.03 mg/mL of each adenosine and inosine from USP Adenosine RS and inosine dissolved in warm water (50 to 55), and diluted with water Standard solution: 0.03 mg/mL of USP Adenosine RS dissolved in warm water (50 to 55) and diluted with water [NOTEIf sodium chloride is present in the Injection, add 0.01 mL of sodium chloride solution (0.9 in 100)/mL of the
62
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USP 32
not to be treated with any toxic, sleep-inducing, or narcosisproducing compounds, and are not to be treated with any compound that would be irritating to the respiratory tract when the Medical Air is used. [NOTEReduce the container pressure by means of a regulator. Measure the gases with a gas volume meter downstream from the detector tube to minimize contamination or change of the specimens.] The various detector tubes called for in the respective tests are listed under Reagents, Indicators, and SolutionsReagent Specifications. LABELING: Where it is piped directly from the collecting tank to the point of use, label each outlet Medical Air.
BACTERIAL ENDOTOXINS TEST 85: When the product is used for rapid intravenous injection, it contains NMT 11.62 USP Endotoxin Units/mg of adenosine. When the product is used for continuous peripheral intravenous infusion, it contains NMT 5.95 USP Endotoxin Units/mg of adenosine. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose containers, preferably of Type I glass, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Adenosine RS USP Endotoxin RS
Alanine
(Comment on this Monograph)id=m1130=Alanine=AMonos.pdf)
Medical Air
(Comment on this Monograph)id=m1000=Medical Air=AMonos.pdf) DEFINITION Medical Air is a natural or synthetic mixture of gases consisting largely of nitrogen and oxygen. It contains NLT 19.5% and NMT 23.5%, by volume, of O2. ASSAY PROCEDURE Analysis: Determine the oxygen concentration of Medical Air using an electrochemical cell analyzer readable to 0.1% of oxygen and calibrated with ambient air to an accuracy of 0.2% of oxygen. [NOTEThe instrument uses the variations of electric current produced by the interaction of oxygen with an electrochemical cell to display the oxygen strength of a confined sample or an in-line flow of the gas. This current generates a signal proportional to the oxygen concentration, which is displayed on a meter.] Acceptance criteria: 19.5%23.5% IMPURITIES Inorganic Impurities CARBON DIOXIDE: Pass 1000 50 mL through a carbon dioxide detector tube at the rate specified for the tube: the indicator change corresponds to NMT 500 ppm. CARBON MONOXIDE: Pass 1000 50 mL through a carbon monoxide detector tube at the rate specified for the tube: the indicator change corresponds to NMT 10 ppm. SULFUR DIOXIDE: Pass 1050 50 mL through a sulfur dioxide detector tube at the rate specified for the tube: the indicator change corresponds to NMT 5 ppm. LIMIT OF NITRIC OXIDE AND NITROGEN DIOXIDE: Pass 550 50 mL through a nitric oxidenitrogen dioxide detector tube at the rate specified for the tube: the indicator change corresponds to NMT 2.5 ppm. WATER AND OIL: Support 1 container in an inverted position (with the valve at the bottom) for 5 min. Cautiously open the valve slightly, maintaining the container in an inverted position. Vent the gas with a barely audible flow against a stainless steel mirror for a few s: no liquid is discernible on the mirror. SPECIFIC TESTS ODOR: Carefully open the container valve to produce a moderate flow of gas. Do not direct the gas stream toward the face, but deflect a portion of the stream toward the nose: no appreciable odor is discernible. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in cylinders or in a lowpressure collecting tank. Containers used for Medical Air are
L-Alanine
C3H7NO2
[56-41-7].
89.09
DEFINITION Alanine contains NLT 98.5% and NMT 101.5% of C3H7NO2, as L-alanine, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 80 mg of Alanine Analysis: Dissolve the Sample in a mixture of glacial acetic acid and formic acid (50:3). Titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 8.909 mg of C3H7NO2. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.15% CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion shows chloride NMT corresponds to 0.70 mL of 0.020 N hydrochloric acid (0.05%). CHLORIDE AND SULFATE, Sulfate 221: A 1.0-g portion shows sulfate NMT corresponds to 0.30 mL of 0.020 N sulfuric acid (0.03%). IRON 241: NMT 30 ppm HEAVY METALS, Method I 231: NMT 15 ppm Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Sample solution: 10 mg/mL of Alanine Standard solution: 0.05 mg/mL of USP L-Alanine RS [NOTEThis solution has a concentration equivalent to 0.5% of that of the Sample solution.] System suitability solution: 0.4 mg/mL each of USP LAlanine RS and USP Glycine RS Spray reagent: 2 mg/mL of ninhydrin in a mixture of butyl alcohol and 2 N acetic acid (19:1) Developing solvent system: Butyl alcohol, glacial acetic acid, and water (3:1:1)
USP 32
Application volume: 5 L Analysis Samples: Sample solution, Standard solution, and System suitability solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. After air-drying the plate, repeat the development process. After air-drying a second time, spray with Spray reagent, and heat to 100105 for 15 min. Examine the plate under white light. The chromatogram obtained from the System suitability solution exhibits two clearly separated spots. Any secondary spot of the Sample solution is not larger or more intense than the principal spot of the Standard solution. Acceptance criteria Individual impurities: NMT 0.5% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +13.7 to +15.1 Sample solution: 100 mg/mL in 6 N hydrochloric acid PH 791: 5.57.0, in a solution (1 in 20) LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 0.2% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP L-Alanine RS USP Glycine RS
Official Monographs / Albendazole 63

necessary. Cool and titrate with 0.1 N perchloric acid VS to a vpotentiometricvUSP32 endpoint v(see Titrimetry 541)vUSP32. Perform a blank determination. Each mL of 0.1 N perchloric acid is equivalent to 26.53 mg of C12H15N3O2S. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Standard stock solution: 5 mg/mL of USP Albendazole RS in glacial acetic acid Standard solution: 0.05 mg/mL of USP Albendazole RS in glacial acetic acid, from Standard solution A Sample solution: 10 mg/mL in glacial acetic acid Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbant: 0.25-mm layer of silica gel Application volume: 10 L Developing solvent system: Chloroform, ether, and glacial acetic acid (60:10:10) Visualization: Short-wavelength UV light Analysis: Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Samples: Standard stock solution, Standard solution, and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved about threefourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the solvent to evaporate from the plate, and examine the plate under short-wavelength UV light. Acceptance criteria: No spot, other than the principal spot of the Sample solution, is larger or more intense than the principal spot of the Standard solution (0.5%). SPECIFIC TESTS LOSS ON DRYING 731: Dry a sample at 105 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Albendazole RS
Albendazole
(Comment on this Monograph)id=m1150=Albendazole=AMonos.pdf)
265.33 C12H15N3O2S Carbamic acid, [5-(propylthio)-1H-benzimidazol-2-yl]-, methyl ester; Methyl 5-(propylthio)-2-benzimidazolecarbamate [54965-21-8]. DEFINITION Albendazole contains NLT 98.0% and NMT 102.0% of C12H15N3O2S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The RF value of the principal spot of the Sample solution corresponds to that of the principal spot of the Standard solution, as obtained in the test for Organic Impurities. ASSAY Change to read: PROCEDURE Sample: 250 mg of Albendazole Analysis: Transfer the Sample to a suitable flask and dissolve in 100 mL of glacial acetic acid, warming gently if
Albendazole Oral Suspension

(Comment on this Monograph)id=m1158=Albendazole Oral Suspension=A-Monos.pdf) DEFINITION Albendazole Oral Suspension is Albendazole in an aqueous vehicle. It contains one or more preservatives and dispersing or suspending agents. It contains NLT 90.0% and NMT 110.0% of the labeled amount of albendazole (C12H15N3O2S). IDENTIFICATION ULTRAVIOLET ABSORPTION 197U Sample stock solution: 1.0 mg/mL of albendazole from a quantity of Suspension, in a mixture of methanol and hydrochloric acid (99:1) [NOTEFilter the mixture, if necessary, to obtain a clear solution.] Sample solution: Sample stock solution and 0.1 N sodium hydroxide (1:99)
64
Albendazole / Official Monographs
USP 32
the Assay with Acidified methanol, prepared as directed for Dissolution, to obtain solutions containing 10 g/mL albendazole. B. The retention time of the major peak for albendazole of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 0.50 g of monobasic ammonium phosphate in 400 mL of water. Add 600 mL of methanol and filter, discarding the first 15 mL of filtrate. Degas the clear filtrate before use. Solution A: Methanol and sulfuric acid (99:1) Internal standard solution: Transfer 150 mg of USP Parbendazole RS to a 50-mL volumetric flask, dissolve with shaking in 5 mL of Solution A and 25 mL of methanol, then dilute with methanol to volume. Standard stock solution: Transfer 100 mg of USP Albendazole RS to a 50-mL volumetric flask, dissolve with shaking in 5 mL of Solution A and 25 mL of methanol, then dilute with methanol to volume. Standard solution: Transfer 5.0 mL of Standard stock solution and 5.0 mL of Internal standard solution to a 50-mL volumetric flask, and dilute with methanol to volume. Sample stock solution: Transfer finely powdered Tablets (NLT 20 Tablets) equivalent to 100 mg of albendazole to a 50-mL volumetric flask, add 5 mL of Solution A and 20 mL of methanol, and shake by mechanical means for about 15 min. Dilute to volume with methanol and filter, discarding the first 15 mL of the filtrate (2 mg/mL). Sample solution: Transfer 5.0 mL of the Sample stock solution and 5.0 mL of the Internal standard solution to a 50mL volumetric flask, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between the albendazole peak and the parbendazole peak Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% for replicate injections Analysis: [NOTEUse peak heights where peak responses are indicated.] Samples: Standard solution and Sample solution Calculate the quantity, in percentage, of C12H15N3O2S in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = peak response ratio of the albendazole peak to the parbendazole peak from the Sample solution = peak response ratio of the albendazole peak to the parbendazole peak from the Standard solution = concentration of USP Albendazole RS in the Standard solution (mg/mL) = nominal concentration of albendazole in the Sample solution (mg/L)
ASSAY PROCEDURE Solution A: Methanol and hydrochloric acid (99:1) Solution B: 13.75 mg/mL of monobasic sodium phosphate Mobile phase: Methanol and Solution B (3:2) Standard stock solution: 1.0 mg/mL of USP Albendazole RS in Solution A Standard solution: 100 g/mL of USP Albendazole RS, from Standard stock solution in Mobile phase Sample stock solution: Equivalent to 1.0 mg/mL of albendazole from a volume of Oral Suspension, in Solution A Sample solution: 100 g/mL of albendazole from Sample stock solution in Mobile phase [NOTEFilter, if necessary, to obtain a clear solution.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 308 nm Column: 4-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H15N3O2S in each mL of the Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Albendazole RS in the Standard solution (mg/mL) = nominal concentration of albendazole in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.55.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Albendazole RS rU rS CS
Albendazole Tablets
(Comment on this Monograph)id=m1162=Albendazole Tablets=A-Monos.pdf) DEFINITION Albendazole Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of albendazole (C12H15N3O2S). IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Solution: Dilute a portion of the clear filtrate used to prepare the Sample solution and a portion of the stock solution used to prepare the Standard solution prepared in
USP 32
Official Monographs / Albumin 65

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Tablets intended for veterinary use only are so labeled. USP REFERENCE STANDARDS 11 USP Albendazole RS USP Parbendazole RS
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min [NOTEDetermine the amount of C12H15N3O2S dissolved using the following Analysis.] Acidified methanol: Methanol and hydrochloric acid (49:1) Standard stock solution: Dissolve 90 mg of USP Albendazole RS in 10 mL of Acidified methanol in a 250-mL volumetric flask with shaking. Dilute with 0.1 N hydrochloric acid to volume. Standard solution: 5.0 mL of the Standard stock solution diluted to 200-mL with 0.1 N sodium hydroxide Sample solution: Sample per Dissolution 711. Transfer 10.0 mL of a filtered portion of the Sample to a 250-mL volumetric flask, and dilute with 0.1 N sodium hydroxide to volume. Blank: 0.1 N sodium hydroxide Analysis: Concomitantly determine the absorbances of the Sample solution and the Standard solution at the wavelengths of maximum and minimum absorbance at about 308 nm and 350 nm against the Blank. Calculate the quantity, in mg, of C12H15N3O2S dissolved: Result = 22.5C(AU/AS) = concentration of USP Albendazole RS in the Standard solution (g/mL) = difference in absorbance between 308 nm and AU 350 nm from the Sample solution = difference in absorbance between 308 nm and AS 350 nm from Standard solution Tolerances: NLT 80% (Q) of the labeled amount is dissolved. UNIFORMITY OF DOSAGE UNITS, Content Uniformity 905 Acidified methanol: Prepare as directed in Dissolution. Standard stock solution: Prepare as directed in Dissolution. Standard solution: Prepare as directed in Dissolution. Sample stock solution: Place 1 Tablet in a 500-mL volumetric flask, add about 300 mL of Acidified methanol, shake by mechanical means for about 30 min, and dilute with Acidified methanol to volume. Filter a portion of this solution, discarding the first 20 mL of the filtrate. Sample solution: Transfer 4.0 mL of the Sample stock solution to a 200-mL volumetric flask, and dilute with 0.1 N sodium hydroxide to volume. Blank: 0.1 N sodium hydroxide Analysis: Concomitantly determine the absorbances of the Standard solution and the Sample solution at the wavelengths of maximum and minimum absorbance at about 308 nm and 350 nm against the Blank. Calculate the quantity, in mg, of C12H15N3O2S in the Tablet taken: Result = 25C(AU/AS) C AU AS = concentration of USP Albendazole RS in the Standard solution (g/mL) = difference in absorbance between 308 nm and 350 nm from the Sample solution = difference in absorbance between 308 nm and 350 nm from Standard solution C
Albumin Human
(Comment on this Monograph)id=m1180=Albumin Human=AMonos.pdf) DEFINITION Albumin Human conforms to the regulations of the federal Food and Drug Administration concerning biologics (640.80 to 640.86) (see Biologics 1041). It is a sterile, nonpyrogenic preparation of serum albumin obtained by fractionating material (source blood, plasma, serum, or placentas) from healthy human donors, the source material being tested for the absence of hepatitis B surface antigen. It is made by a process that yields a product that is safe for intravenous use. NLT 96% of its total protein is albumin. It is a solution containing, in each 100 mL, either 25 g of serum albumin osmotically equivalent to 500 mL of normal human plasma, or 20 g equivalent to 400 mL, or 5 g equivalent to 100 mL, or 4 g equivalent to 80 mL thereof, and contains NLT 93.75% and NMT 106.25% of the labeled amount in the case of the solution containing 4 g in each 100 mL, and NLT 94.0% and NMT 106.0% of the labeled amount in the other cases. It contains no added antimicrobial agent, but may contain sodium acetyltryptophanate with or without sodium caprylate as a stabilizing agent. It has a sodium content of NLT 130 mEq /L and NMT 160 mEq/L. It has a heme content such that the absorbance of a solution, diluted to contain 1% of protein, in a 1-cm holding cell, measured at a wavelength of 403 nm, is NMT 0.25. It meets the requirements of the test for heat stability and for pH. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at the temperature recommended by the manufacturer or indicated on the label. EXPIRATION DATE: The expiration date is not later than 5 years after issue from manufacturers cold storage (5, 3 years) if labeling recommends storage between 2 and 10; not later than 3 years after issue from manufacturers cold storage (5, 3 years) if labeling recommends storage at temperatures not higher than 37; and not later than 10 years after date of manufacture if in a hermetically sealed metal container and labeling recommends storage between 2 and 10. LABELING: Label it to state that it is not to be used if it is turbid and that it is to be used within 4 h after the container is entered. Label it also to state the osmotic equivalent in terms of plasma, the sodium content, and the type of source material (venous plasma, placental plasma, or both) from which it was prepared. Label it also to indicate that additional fluids are needed when the 20-g/100-mL or 25g/100-mL product is administered to a markedly dehydrated patient.
66
Albuterol / Official Monographs

SPECIFIC TESTS WATER DETERMINATION, Method I 921:
USP 32
Albuterol
(Comment on this Monograph)id=m1195=Albuterol=AMonos.pdf)
NMT 0.5%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Albuterol RS
C13H21NO3 239.31 1,3-Benzenedimethanol, 1-[[(1,1dimethylethyl)amino]methyl]-4-hydroxy-; 1-[(tert-Butylamino)methyl]-4-hydroxy-m-xylene-,-diol [18559-94-9]. DEFINITION Albuterol contains NLT 98.5% and NMT 101.0% of C13H21NO3, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 80 g/mL in 0.1 N hydrochloric acid ASSAY PROCEDURE Sample solution: 8 mg/mL of Albuterol in glacial acetic acid Analysis: To 50 mL of Sample solution, add 2 drops of crystal violet TS, and titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 23.93 mg of C13H21NO3. Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Standard solution: 0.10 mg/mL of USP Albuterol RS in methanol Sample solution: 20 mg/mL of Albuterol in methanol Chromatographic system See Chromatography 621, Thin-Layer Chromatography Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 10 L Developing solvent system: Methyl isobutyl ketone, isopropyl alcohol, ethyl acetate, ammonium hydroxide, and water (50:45:35:3:18) Visualization: Iodine vapor Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography, applying aliquots of the Standard solution and the Sample solution. Develop in the Developing solvent system until the solvent front has moved three-fourths the length of the plate. Remove the plate from the developing chamber, air-dry, and expose it to iodine vapor. Acceptance criteria: Any spot, other than the principal spot, obtained from the Sample solution is not greater in size and intensity than the spot produced by the Standard solution (0.5%), and the sum of the impurities is not greater than 2.0%.
Albuterol Sulfate
(Comment on this Monograph)id=m1210=Albuterol Sulfate=AMonos.pdf) 576.70 (C13H21NO3)2 H2SO4 1,3-Benzenedimethanol, 1-[[(1,1dimethylethyl)amino]methyl]-4-hydroxy-, sulfate (2:1) (salt); 1-[(tert-Butylamino)methyl]-4-hydroxy-m-xylene-,-diol sulfate (2:1) (salt) [51022-70-9]. DEFINITION Albuterol Sulfate contains NLT 98.5% and NMT 101.0% of (C13H21NO3)2 H2SO4, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 80 g/mL in 0.1 N hydrochloric acid C. IDENTIFICATION TESTSGENERAL, Sulfate 191 Sample solution: Shake an amount of sample equivalent to 4 mg of albuterol with 10 mL of water, and filter. Acceptance criteria: Meets the requirements of the tests D. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 3.85 mg/mL of ammonium acetate Mobile phase: Isopropanol, Solution A, and water [(5 1):30:65], filtered and degassed. Adjust dropwise with acetic acid to a pH of 4.5 0.3. Standard solution: 0.6 mg/mL of USP Albuterol Sulfate RS Sample solution: 0.6 mg/mL of Albuterol Sulfate Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 276 nm Column: 4.6-mm 20-cm; packing L10 Flow rate: 2.0 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.5 between albuterol and 4-[2-[(1,1dimethylethyl)amino]-1-hydroxyethyl]-2-methylphenol sulfate Relative standard deviation: NMT 1.5% Analysis: Sample: Standard solution and Sample solution Calculate the quantity, in percentage, of (C13H21NO3)2 H2SO4 in the portion of Albuterol Sulfate taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
USP 32
= concentration of USP Albuterol Sulfate RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: NLT 98.5% and NMT 101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Acceptance criteria: NMT 0.1% Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 0.10 mg/mL of USP Albuterol Sulfate RS Sample solution: 20 mg/mL of Albuterol Sulfate Application volume: 10 L Developing solvent system: Methyl isobutyl ketone, isopropyl alcohol, ethyl acetate, ammonium hydroxide and water (50:45:35:3:18) Visualization: Iodine vapor Analysis: Proceed as directed in Chromatography 621, Thin-layer Chromatography, applying aliquots of the Standard solution and the Sample solution. Develop in the Developing solvent system until the solvent front has moved three-fourths the length of the plate. Remove the plate from the developing chamber, air-dry, and expose it to iodine vapor. Acceptance criteria: Any spot, other than the principal spot, obtained from the Sample solution is not greater in size and intensity than the spot produced by the Standard solution (0.5%), and the sum of the impurities is not greater than 2.0%. SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5% CS
Official Monographs / Albuterol 67

Diluent: Methanol and water (2:3) Mobile phase: Methanol and Solution B (2:3) Standard stock solution: 0.12 mg/mL of USP Albuterol Sulfate RS in a mixture of Solution A and methanol [NOTETo USP Albuterol Sulfate RS, add a portion of Solution A corresponding to 60% of the final solution volume. Sonicate for 5 min, and dilute to final volume with methanol.] Standard solution: 0.03 mg/mL of USP Albuterol Sulfate RS from Standard stock solution, in Diluent Sample solution: To the whole Tablets, add a portion of Solution A corresponding to 60% of the final solution volume, shake by mechanical means for 45 min, sonicate for 10 min, allow to cool to room temperature, and dilute to final volume with methanol equivalent to 25 g/mL of albuterol from a number of whole Tablets in a mixture of Solution A and methanol. Pass through a suitable 0.45-m or finer porosity fliter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 276 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 800 theoretical plates determined from the analyte peak Tailing factor: NMT 2.5 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample Solution Calculate the quantity, as a percentage, of C13H21NO3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) N (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Albuterol Sulfate RS in the Standard solution (mg/mL) CU = nominal concentration of albuterol in the Sample solution (mg/mL) N = number of molecules of albuterol released from each molecule of albuterol sulfate, 2 = molecular weight of albuterol, 239.31 Mr1 Mr2 = molecular weight of albuterol sulfate, 576.70 Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711, Procedure for a Pooled Sample Medium: Water; 500 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of C13H21NO3 dissolved using the following method. [NOTEFor the Mobile phase, Standard solution, and Chromatographic system, prepare as directed in the Assay.] Analysis: Inject a suitable volume (about 100 L) of a portion of the Sample solution, previously passed through a 0.45-m nylon filter, into the chromatograph. Calculate the quantity of C13H21NO3 dissolved by comparing this peak response with the major peak response similarly obtained on chromatographing the Standard solution previously diluted, if necessary, with a mixture of water and methanol (6:4) to
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Albuterol Sulfate RS
Albuterol Tablets
(Comment on this Monograph)id=m1218=Albuterol Tablets=AMonos.pdf) DEFINITION Albuterol Tablets contain an amount of albuterol sulfate [(C13H21NO3)2 H2SO4] equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of albuterol (C13H21NO3). IDENTIFICATION A. The RF value of the principal spot obtained from the Sample solution corresponds to that obtained from Standard solution A obtained as directed in the Procedure for Organic Impurities. B. IDENTIFICATION TESTSGENERAL, Sulfate 191: Shake a quantity of the powdered tablets equivalent to 4 mg of albuterol with 10 mL, and filter. The filtrate so obtained meets the requirements of the test. ASSAY PROCEDURE Solution A: Dilute 20 mL of glacial acetic acid to 2 L. Solution B: 1.13 g of sodium 1-hexanesulfonate in 1200 mL of water. Add 12 mL of glacial acetic acid.
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Albuterol / Official Monographs
USP 32
obtain a Standard solution having a known concentration of USP Albuterol Sulfate RS approximately corresponding to the concentration of the Sample solution. Tolerances: NLT 80% (Q) of the labeled amount of C13H21NO3 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution A: Prepare a solution of USP Albuterol Sulfate RS having a known concentration of 0.580 mg/mL equivalent to 0.483 mg of albuterol. Standard solution B: Prepare a solution of USP Albuterol Sulfate RS having a known concentration of 0.218 mg/mL equivalent to 0.183 mg of albuterol. Standard solution C: Prepare a solution of USP Albuterol Sulfate RS having a known concentration of 0.073 mg/mL equivalent to 0.061 mg of albuterol. Sample solution: Place a quantity of finely powdered Tablets, equivalent to 48 mg of albuterol, into a suitable container. Add 60 mL of diluted alcohol (1 in 2), and shake by mechanical means for 30 min. Filter the mixture, and wash the filter with small portions of alcohol, combining this with the filtrate. Evaporate the filtrate to dryness under reduced pressure at a temperature below 40. Dissolve the residue as completely as possible in 2 mL. Application volume: 10-L aliquots (in two successive portions of 5 L, allowing the solvent to evaporate between applications) Developing solvent system: Methyl isobutyl ketone, isopropyl alcohol, ethyl acetate, ammonium hydroxide, and water (50:45:35:3:18) Spray reagent A: 3-Methyl-2-benzothiazolinone hydrazone hydrochloride TS Spray reagent B: Ammoniacal potassium ferricyanide TS Analysis Samples: Sample solution, Standard solution A, Standard solution B, and Standard solultion C Proceed as directed for Chromatography 621, Thin-layer Chromatography. Develop the chromatograms until the solvent front has moved about 17 cm. Spray the plate first with Spray reagent A, and then Spray reagent B, and finally again with Spray reagent A. Examine the plate and estimate the responses of any secondary spots observed in the lane of the Sample solution by comparison with those of Standard solutions A, B, and C. Acceptance criteria: No major secondary spot is greater in size or intensity than the principal spot produced by Standard solution A (2.0%). No other secondary spot is greater in size or intensity than the principal spot produced by Standard solution B (0.75%). No more than two other secondary spots are equal in size or intensity than the principal spot produced by Standard solution C (0.25%). The sum of the intensities of all secondary spots obtained from the Sample solution corresponds to NMT 3.5%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Albuterol Sulfate RS
Alclometasone Dipropionate
(Comment on this Monograph)id=m1225=Alclometasone Dipropionate=A-Monos.pdf)
C28H37ClO7 521.04 Pregna-1,4-diene-3,20-dione, 7-chloro-11-hydroxy-16methyl-17,21-bis(1-oxopropoxy)-, (7,11,16)-; 7-Chloro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 17,21-dipropionate [66734-13-2]. DEFINITION Alclometasone Dipropionate contains NLT 97.0% and NMT 102.0% of C28H37ClO7, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the Internal standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 6.80 mg/mL of monobasic potassium phosphate (0.05 M) Mobile phase: Methanol and Solution A (2:1) Internal standard solution: 2 mg/mL of betamethasone dipropionate in methanol Standard stock solution: 1.2 mg/mL of USP Alclometasone Dipropionate RS in methanol Standard solution: 4.0 mL Standard stock solution and 4.0 mL Internal standard solution. Dilute with methanol to 25 mL. [NOTEThis solution contains approximately 0.2 mg/mL of USP Alclometasone Dipropionate RS.] Sample stock solution: 1.2 mg/mL of Alclometasone Dipropionate in methanol Sample solution: 4 mL Sample stock solution and 4 mL Internal standard solution. Dilute with methanol to 25 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for alclometasone dipropionate and betamethasone dipropionate are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the analyte and the Internal standard solution peaks
USP 32
Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C28H37ClO7 in the portion taken: Result = (rU/rS) (CS/CU) 100 = peak height ratio from the Sample solution = peak height ratio from the Standard solution = concentration of USP Alclometasone Dipropionate RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 30 ppm Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Diluent: Dilute 250 mL methanol with dichloromethane to 500 mL. Standard stock solution: 250 10 g/mL of USP Alclometasone Dipropionate RS in Diluent Standard solutions: Dilute volumes of Standard stock solution quantitatively with Diluent to obtain Standard solutions, designated below by letter, having the following concentrations.
Standard Solution A B C D E F Dilution (4 in 5) (3 in 5) (2 in 5) (3 in 10) (2 in 10) (1 in 10) Concentration (g/mL) 200 150 100 75 50 25 Percentage (%) 2.0 1.5 1.0 0.75 0.50 0.25
Official Monographs / Alclometasone 69

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Alclometasone Dipropionate RS
rU rS CS
Alclometasone Dipropionate Cream

(Comment on this Monograph)id=m1227=Alclometasone Dipropionate Cream=A-Monos.pdf) DEFINITION Alclometasone Dipropionate Cream contains NLT 90.0% and NMT 110.0% of the labeled amount of alclometasone dipropionate (C28H37ClO7) in a suitable cream base. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the Internal standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 0.08 mg/mL in methanol Sample solution: Place a quantity of Cream, equivalent to 1.25 mg of alclometasone dipropionate, in a 50-mL centrifuge tube, and add 15 mL of methanol. Insert a stopper securely into the tube, and place the tube in a water bath maintained at 60 until the semisolid components melt. Remove the tube from the bath, shake vigorously until the specimen components resolidify, and place the tube in an icemethanol bath for 15 min. Remove the tube from the bath, and centrifuge at 2500 rpm for 5 min. Transfer the clear supernatant to a vial, and allow this Sample solution to equilibrate to room temperature. Application volume: 20 L Visualization: Short-wavelength UV light Developing solvent system: Chloroform and acetone (7:1) Analysis: Standard solution and Sample solution [NOTEDry the applications with the aid of a stream of nitrogen.] Proceed as directed for Chromatography 621, Thin-Layer Chromatography until the solvent has moved about threefourths the length of the plate. Observe the plate under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot obtained from the Sample solution corresponds to that obtained from the Standard solution. ASSAY PROCEDURE Solution A: 6.80 mg/mL of monobasic potassium phosphate (0.05 M) Mobile phase: Methanol and Solution A (2:1) Internal standard solution: 0.4 mg/mL of betamethasone dipropionate in methanol Standard stock solution: 0.25 mg/mL of USP Alclometasone Dipropionate RS in methanol Standard solution: Transfer about 25 mg of USP Alclometasone Diproprionate RS to a 100-mL volumetric flask, add methanol to volume, and mix. Transfer 5.0 mL of this solution to a small stoppered flask, add 5.0 mL of methanol and 5.0 mL of Internal standard solution, and mix to obtain a Standard solution having a known concentration of about 0.08 mg of USP Alclometasone Dipropionate RS per mL. Sample solution: Transfer a quantity of Cream, equivalent to 1.25 mg of alclometasone dipropionate, to a 50-mL
Sample solution: 10 0.4 mg/mL of Alclometasone Dipropionate in Diluent Application volume: 25 L Visualization: Short-wavelength UV light Developing solvent system: Chloroform and acetone (7:1) Analysis Samples: Standard solutions AF and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatograms until the solvent front has moved about three-fourths the length of the plate. Observe the plate under short-wavelength UV light, and compare the intensities of any secondary spots observed in the chromatogram of the Sample solution with those of the principal spots in the chromatograms of the Standard solutions. Acceptance criteria: The sum of the intensities of secondary spots obtained from the Sample solutions corresponds to NMT 3.0% of related compounds. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +21 to +25 Sample solution: 30 mg/mL in dioxane LOSS ON DRYING 731: Dry it in vacuum at a pressure not exceeding 5 mm of mercury at 105 for 3 h: it loses NMT 0.5% of its weight.
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Alclometasone / Official Monographs
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relative to the Internal standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 0.25 mg/mL USP Alclometasone Dipropionate RS in methanol Sample solution: Place a quantity of Ointment, equivalent to 1.25 mg of alclometasone dipropionate, in a 50-mL centrifuge tube, add 10 mL of 2,2,4-trimethylpentane, insert a stopper securely into the tube, and disperse the specimen using a vortex mixer. Add 5.0 mL of a solution of methanol in water (45 in 50), insert the stopper securely, shake vigorously for 2 min, and centrifuge at 2500 rpm for 3 min. Remove the lower, aqueous alcohol phase, and transfer this Sample solution to a stoppered vial. Application volume: 20 L Developing solvent system: Chloroform and acetone (7:1) Analysis Samples: Standard solution and Sample solution [NOTEDry the applications with the aid of a stream of nitrogen.] Proceed as directed for Chromatography 621, Thin-Layer Chromatography until the solvent front has moved about three-fourths of the length of the plate. Observe the plate under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot obtained from the Sample solution corresponds to that obtained from the Standard solution. ASSAY PROCEDURE Solution A: 6.80 mg/mL of monobasic potassium phosphate (0.05 M) Solution B: Dilute 450 mL of methanol to 500 mL with water. Mobile phase: Methanol and Solution A (2:1) Internal standard solution: 0.15 mg/mL of betamethasone dipropionate in Solution B Standard stock solution: 0.1 mg/mL of USP Alclometasone Dipropionate RS in Solution B Standard solution: 0.05 mg/mL of USP Alclometasone Dipropionate RS obtained from a Standard stock solution and Internal standard solution (1:1) Sample solution: Transfer a quantity of Ointment, equivalent to 0.5 mg of alclometasone dipropionate, to a 50-mL centrifuge tube, add 10 mL of 2,2,4trimethylpentane, insert a stopper securely into the tube, and disperse the specimen using a vortex mixer. Add 5.0 mL of Internal standard solution and 5.0 mL of Solution B, insert the stopper securely, shake vigorously for 2 min, and centrifuge at 2500 rpm for 3 min. Remove the lower, aqueous alcohol phase, and transfer this Sample solution to a stoppered vial. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for alclometasone dipropionate and betamethasone dipropionate are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the analyte and internal standard peaks
centrifuge tube. Add 5.0 mL of Internal standard solution, and add 10.0 mL of methanol. Insert a stopper securely into the tube, and place it in a water bath maintained at 60 until the semisolid components melt. Remove the tube from the bath, shake vigorously until the specimen components resolidify, and return the tube to the 60 water bath until the semisolid components melt. Remove the tube from the bath, shake vigorously until the specimen components resolidify, and place the tube in an icemethanol bath for 15 min. Remove the tube from the bath, and centrifuge at 2500 rpm for 5 min. Transfer the clear supernatant to a small stoppered flask, and allow this Sample solution to equilibrate to room temperature. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for alclometasone dipropionate and betamethasone dipropionate are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the analyte and internal standard peaks Relative standard deviation: NMT 2% Analysis: Standard solution and Sample solution Calculate the quantity, as a percentage, of C28H37ClO7 in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 = peak height ratio from the Sample solution = peak height ratio from the Standard solution = concentration of USP Alclometasone Dipropionate RS in the Standard solution (mg/mL) = nominal concentration of alclometasone CU dipropionate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Alclometasone Dipropionate RS RU RS CS
Alclometasone Dipropionate Ointment

(Comment on this Monograph)id=m1230=Alclometasone Dipropionate Ointment=A-Monos.pdf) DEFINITION Alclometasone Dipropionate Ointment contains NLT 90.0% and NMT 110.0% of the labeled amount of alclometasone dipropionate (C28H37ClO7), in a suitable ointment base. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both
USP 32
Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Calculate the quantity, as a percentage, of C28H37ClO7 in the portion of Ointment taken: Result = (RU/RS) (CS/CU) 100 = peak height ratio from the Sample solution = peak height ratio from the Standard solution = concentration of USP Alclometasone Dipropionate RS in the Standard solution (mg/mL) = nominal concentration of alclometasone CU dipropionate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU RS CS SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61, and TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Alclometasone Dipropionate RS
Official Monographs / Alcohol 71

examined. Dilute 100 L of the solution to 10.0 mL with Sample solution A. Standard solution C: Dilute 150 L of acetal to 50.0 mL with the substance to be examined. Dilute 100 L of the solution to 10.0 mL with Sample solution A. Standard solution D: Dilute 100 L of benzene to 100.0 mL with Sample solution A. Dilute 100 L of this solution to 50.0 mL with Sample solution A. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.32-mm 30-m fused silica capillary column bonded with a 1.8-m layer of phase G43 Split ratio: 1:20 Temperature Column:
Time (min) 0 12 32 42 Temperature () 40 40 240 240
Alcohol
(Comment on this Monograph)id=m1238=Alcohol=AMonos.pdf)
C2H6O Ethanol; Ethyl alcohol [64-17-5].
46.07
DEFINITION Alcohol contains NLT 92.3% and NMT 93.8%, by weight, corresponding to NLT 94.9% and NMT 96.0%, by volume, at 15.56, of C2H5OH. IDENTIFICATION A. PROCEDURE: It meets the requirements of the test for Specific Gravity 841. B. INFRARED ABSORPTION 197F or 197S: Neat IMPURITIES Inorganic Impurities LIMIT OF NONVOLATILE RESIDUE: Evaporate 100 mL in a tared dish on a water bath, and dry at 100 to 105 for 1 h: the weight of the residue is NMT 2.5 mg. Organic Impurities PROCEDURE: VOLATILE IMPURITIES Sample solution A: Substance to be examined Sample solution B: Add 150 L of 4-methylpentan-2-ol to 500.0 mL of the substance to be examined. Standard solution A: Dilute 100 L of methanol to 50.0 mL with the substance to be examined. Dilute 5.0 mL of the solution to 50.0 mL with Sample solution A. Standard solution B: Dilute 50 L of methanol and 50 L of acetaldehyde to 50.0 mL with the substance to be
Detector: 280 Injection port: 200 Linear velocity: 35 cm/min Carrier gas: Helium Injection size: 1.0 L System suitability Sample: Standard solution B Suitability requirements Resolution: Between the first major peak (acetaldehyde) and the second major peak (methanol) is NLT 1.5 in the Standard solution B Analysis Samples: Standard solution A, Standard solution C, Standard solution D, Sample solution A and Sample solution B Calculate the concentration of methanol in Sample solution A: NMT half the area of the corresponding peak in the chromatogram obtained with Standard solution A (200 ppm). Calculate the sum of the contents of acetaldehyde and acetal (ppm), expressed as acetaldehyde: Result (ppm) = (10 AE)/(AT AE) + (30 CE)/(CT CE) = area of the acetaldehyde peak obtained with Sample solution A = area of the acetaldehyde peak obtained with AT Standard solution B = area of the acetal peak obtained with Sample CE solution A CT = area of the acetal peak obtained with Standard solution C: NMT 10 ppm, expressed as acetaldehyde Calculate the content of benzene (ppm): AE Result (ppm) = (2BE)/(BT BE) = area of the benzene peak obtained with Sample solution A = area of the benzene peak obtained with BT Standard solution D: NMT 2 ppm If necessary, the identity of benzene can be confirmed using another suitable chromatographic system (stationary phase with a different polarity). BE
72
Alcohol / Official Monographs

The total of all other impurities in the chromatogram obtained with Sample solution B: NMT area of the peak due to 4-methylpentan-2-ol in the chromatogram obtained with Sample solution B (300 ppm). Disregard any peaks that are 0.03 times the area of the peak corresponding to 4-methylpentan-2-ol in the chromatogram obtained with Sample solution B (9 ppm).
USP 32
ACIDITY OR ALKALINITY Phenolphthalein solution: 1 mg/mL of phenolphthalein in a mixture of alcohol and water (4:1) Analysis: To 20 mL of alcohol, add 20 mL of freshly boiled and cooled water and 0.1 mL of Phenolphthalein solution. The solution is colorless. Add 1.0 mL of 0.01 N sodium hydroxide. The solution is pink (30 ppm, expressed as acetic acid). COLOR OF SOLUTION Standard stock solution: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (3:3:2.4:1.6) (10 g/L) Standard solution: Transfer 1.0 mL of Standard stock solution to a 100-mL volumetric flask, and dilute with dilute hydrochloric acid (10 g/L). [NOTEPrepare the Standard solution immediately before use.] Sample solution: Substance to be examined Analysis: Transfer a sufficient portion of the Sample solution to a test tube of colorless, transparent, neutral glass with a flat base and an internal diameter of 15 to 25 mm to obtain a depth of 40 mm. Similarly transfer portions of Standard solution and water to separate, matching test tubes. Compare the Sample solution, Standard solution, and water in diffused daylight, viewing vertically against a white background. (See Spectrophotometry and Light-Scattering 851, Visual Comparison.) Acceptance criteria: The Sample solution has the appearance of water or is not more intensely colored than the Standard solution. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. USP REFERENCE STANDARDS 11 USP Alcohol RS
SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.8120.816 at 15.56, indicating 92.3%93.8%, by weight, or 94.9%96.0%, by volume, of C2H5OH ULTRAVIOLET ABSORPTION Analytical wavelength: 200 to 400 nm Cell: 5 cm Acceptance criteria: Maximum absorbance 0.40 at 240 nm, 0.30 between 250 and 260 nm, and 0.10 between 270 and 340 nm. Examine between 235 and 340 nm, using water as the compensation liquid. The absorption curve is smooth. CLARITY OF SOLUTION [NOTEThe Sample solution is to be compared to Reference suspension A and to water in diffused daylight 5 min after preparation of Reference suspension A.] Hydrazine solution: 10 mg/mL of hydrazine sulfate in water [NOTEAllow to stand for 4 to 6 h.] Methenamine solution: Transfer 2.5 g of methenamine to a 100-mL glass-stoppered flask, add 25.0 mL of water, insert the glass stopper, and mix to dissolve. Primary opalescent suspension: Transfer 25.0 mL of Hydrazine solution to the Methenamine solution in the 100mL glass-stoppered flask. Mix, and allow to stand for 24 h. [NOTEThis suspension is stable for 2 months, provided it is stored in a glass container free from surface defects. The suspension must not adhere to the glass and must be well mixed before use.] Opalescence standard: Transfer 15.0 mL of the Primary opalescent suspension to a 1000-mL volumetric flask, and dilute with water to volume. [NOTEThis suspension should not be used beyond 24 h after preparation.] Reference suspension A: Transfer 5.0 mL of the Opalescence standard to a 100-mL volumetric flask, and dilute with water to volume. Reference suspension B: Transfer 10.0 mL of the Opalescence standard to a second 100-mL volumetric flask, and dilute with water to volume. Sample solution A: Substance to be examined Sample solution B: Dilute 1.0 mL of Sample solution A to 20 mL with water, and allow to stand for 5 min before testing. Analysis: Transfer a sufficient portion of Sample solution A and Sample solution B to separate test tubes of colorless, transparent, neutral glass with a flat base and an internal diameter of 15 to 25 mm to obtain a depth of 40 mm. Similarly transfer portions of Reference suspension A, Reference suspension B, and water to separate matching test tubes. Compare Sample solution A, Sample solution B, Reference suspension A, Reference suspension B, and water in diffused daylight, viewing vertically against a black background. (See Spectrophotometry and Light-Scattering 851, Visual Comparison.) [NOTEThe diffusion of light must be such that Reference suspension A can readily be distinguished from water, and that Reference suspension B can readily be distinguished from Reference suspension A.] Acceptance criteria: Sample solution A and Sample solution B show the same clarity as that of water or their opalescence is not more pronounced than that of Reference suspension A.
Dehydrated Alcohol
(Comment on this Monograph)id=m1240=Dehydrated Alcohol=A-Monos.pdf)
C2H6O Ethanol; Ethyl alcohol [64-17-5]. DEFINITION Dehydrated Alcohol contains NLT 99.2%, by weight, corresponding to NLT 99.5%, by volume, at 15.56, of C2H5OH. IDENTIFICATION A. SPECIFIC GRAVITY 841: NMT 0.7962 at 15.56, indicating NLT 99.2% of C2H5OH, by weight B. INFRARED ABSORPTION 197S or 197F: Neat SPECIFIC TESTS ULTRAVIOLET ABSORPTION (See Spectroscopy and Light-Scattering 851
46.07
USP 32
Analytical wavelength: 235340 nm Cell: 5 cm Blank: Water Acceptance criteria Absorbance: NMT 0.40 at 240 nm; NMT 0.30 between 250 and 260 nm; and NMT 0.10 between 270 and 340 nm [NOTEThe absorption curve is smooth.] LIMIT OF NONVOLATILE RESIDUE Sample: 100 mL of Dehydrated Alcohol Analysis: Evaporate the sample in a tared dish on a water bath, and dry at 100105 for 1 h. Acceptance criteria: The weight of the residue is NMT 2.5 mg. CLARITY OF SOLUTION [NOTEThe Sample solution is to be compared to Reference suspension A and to water in diffused daylight 5 min after preparation of Reference suspension A.] Hydrazine solution: 10 mg/mL of hydrazine sulfate [NOTEAllow to stand for 46 h.] Reference suspension: 2.5 g of Methenamine and add 25.0 mL to a 100-mL glass-stoppered flask. Insert the glassstopper, and mix to dissolve. Add 25.0 mL of Hydrazine solution, mix, and allow to stand for 24 h. [NOTEThis suspension is stable for 2 months, provided it is stored in a glass container free from surface defects. The suspension must not adhere to the glass and must be well mixed before use.] Standard stock suspension: Dilute 15.0 mL of Reference suspension to 1000 mL. [NOTEThis suspension should not be used beyond 24 h after preparation.] Standard suspension A: Dilute 5.0 mL of Standard stock suspension to 100 mL. Standard suspension B: Dilute 10.0 mL of Standard stock suspension to 100 mL. Sample solution A: Substance to be examined Sample solution B: 1.0 mL of Sample solution A diluted to 20 mL Blank: Water. [NOTEAllow to stand for 5 min before testing.] Analysis Samples: Sample solution A, Sample solution B, Standard suspension A, Standard suspension B, and Blank. Transfer a sufficient portion of Sample solution A and Sample solution B to separate test tubes of colorless, transparent, neutral glass with a flat base and an internal diameter of 1525 mm to obtain a depth of 40 mm. Similarly transfer portions of Standard suspension A, Standard suspension B, and Blank to separate matching test tubes. Compare samples in diffused daylight, viewing vertically against a black background (see Spectrophotometry and Light-Scattering 851, Visual Comparison). Acceptance criteria: Sample solution A and Sample solution B show the same clarity as that of water, or their opalescence is not more pronounced than that of Standard suspension A. [NOTEThe diffusion of light must be such that Standard suspension A can readily be distinguished from water, and that Standard suspension B can readily be distinguished from Standard suspension A.] ACIDITY OR ALKALINITY Phenolphthalein solution: Dissolve 0.1 g of phenolphthalein in 80 mL alcohol, and dilute to 100 mL with water. Analysis: To 20 mL of Dehydrated Alcohol add 20 mL of freshly boiled and cooled water and 0.1 mL of phenolphthalein solution. The solution is colorless. Add 1.0 mL of 0.01 N sodium hydroxide. Acceptance criteria: The solution is pink (30 ppm, expressed as acetic acid).

COLOR OF SOLUTION Standard stock solution: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (10 mg/mL) (3:3:2.4:1.6) Standard solution: 1.0 mL of Standard stock solution, and dilute to 100 mL with dilute hydrochloric acid (10 mg/mL) [NOTEPrepare this immediately before use.] Sample solution: Substance to be examined Analysis Samples: Sample solution,Standard solution, and water. Transfer a sufficient portion of each Sample solution to individual test tubes of colorless, transparent, neutral glass with a flat base and an internal diameter of 1525 mm to obtain a depth of 40 mm. Compare the Samples in diffused daylight, viewing vertically against a white background (see Spectrophotometry and Light-Scattering 851, Visual Comparison). Acceptance criteria: The Sample solution has the appearance of water or is not more intensely colored than the Standard solution. VOLATILE IMPURITIES Sample solution A: Substance to be examined Sample solution B: 150 L of 4-methylpentan-2-ol to 500.0 mL of the substance to be examined Standard solution A: 0.2 L/mL of methanol in Sample solution A. Standard solution B: 0.01 L/mL of methanol and 0.01 L/mL of acetaldehyde in Sample solution A. Standard solution C: 0.03 L/mL of acetal in Sample solution A. Standard solution D: 2 nL/mL of benzene in Sample solution A. Chromatographic system Mode: GC Detector: Flame ionization Column: 0.32-mm 30-m fused silica capillary column bonded with a 1.8-m layer of phase G43 Split ratio: 1:20 Temperature: See the temperature program table below.
Time (min) Injection port Detector port Column 0 12 32 42 Temperature 200 280 40 40 240 240
Flow rate: 35 cm/s Carrier gas: Helium Injection size: 1 L System suitability Sample: Standard solution B Suitability requirements Resolution: NLT 1.5 between the first major peak (acetaldehyde) and the second major peak (methanol) Analysis 1 Samples: Sample solution A, Sample solution B, Standard solution A, Standard solution B, Standard solution C, and Standard Solution D Acceptance criteria 1: The area of the methanol peak from Sample solution A is NMT half the area of the corresponding peak from Standard solution A (200 ppm).
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Analysis 2 Calculate the sum of the contents of acetaldehyde and acetal, expressed as acetaldehyde, using the following formula: Result = (10 AE)/(AT AE) + (30 CE)/(CT CE) = area of the acetaldehyde peak from Sample solution A = area of the acetaldehyde peak from Standard AT solution B = area of the acetal peak from Sample solution A CE = area of the acetal peak from Standard solution C CT Acceptance criteria 2: NMT 10 ppm, expressed as acetaldehyde Analysis 3 Calculate the content of benzene using the following formula: AE Result = (2BE)/(BT BE)
USP 32
IMPURITIES Organic Impurities METHANOL Analysis: To 1 drop of sample add 1 drop of dilute phosphoric acid (1 in 20) and 1 drop of 50 g/mL potassium permanganate solution. Mix, allow to stand for 1 min, and add 50 g/mL sodium metabisulfite solution, dropwise, until the permanganate color is discharged. If a brown color remains, add 1 drop of dilute phosphoric acid (1 in 20). To the colorless solution, add 5 mL of freshly prepared chromotropic acid TS, and heat in a water bath at 60 for 10 min. Acceptance criteria: No violet color appears. ALDEHYDES AND OTHER FOREIGN ORGANIC SUBSTANCES [NOTEAll glassware used in this test should be thoroughly cleaned with hydrochloric acid, then rinsed with water and finally with a volume of dehydrated alcohol injection.] Sample: 20 mL Analysis: Place the Sample in a glass-stoppered cylinder, cool the contents to approximately 15, and add, by pipet, 0.10 mL of 0.10 N potassium permanganate, noting accurately the time of addition. Mix at once by inverting the stoppered cylinder, and allow it to stand at 15 for 5 min. Acceptance criteria: The pink color does not entirely disappear. LIMIT OF ACETONE AND ISOPROPYL ALCOHOL Sample: 1.0 mL Solution A: 1 g/mL of furfural Analysis: To the Sample add 1 mL of water, 1 mL of a saturated solution of dibasic sodium phosphate, and 3 mL of a saturated solution of potassium permanganate. Warm the mixture to 4550, and allow to stand until the permanganate color is discharged. Add 3 mL of 2.5 N sodium hydroxide, and pass, without washing, through a sintered-glass filter. Prepare a control containing 1 mL of the saturated solution of dibasic sodium phosphate, 3 mL of 2.5 N sodium hydroxide, and 80 g of acetone in 9 mL. To each solution, add 1 mL of Solution A, and allow to stand for 10 min, then to 1.0 mL of each solution, add 3 mL of hydrochloric acid. Acceptance criteria: Any pink color produced by the Sample is not more intense than that produced by the control. SPECIFIC TESTS SPECIFIC GRAVITY 841 Acceptance criteria: NMT 0.8035 at 15.56, indicating NLT 96.8%, by weight, of C2H5OH ACIDITY Sample: 50 mL Analysis: To the Sample, in a glass-stoppered flask, add 50 mL of recently boiled water, phenolphthalein TS, and titrate with 0.020 N sodium hydroxide to a pink color that persists for 30 s. Acceptance criteria: NMT 10.0 mL of 0.020 N sodium hydroxide is required. AMYL ALCOHOL AND NONVOLATILE, CARBONIZABLE SUBSTANCES Sample: 25 mL Analysis: Allow the Sample to evaporate spontaneously from a porcelain dish, carefully protected from dust, until the surface of the dish is barely moist. Acceptance criteria: No red or brown color is produced immediately upon the addition of a few drops of sulfuric acid. LIMIT OF NONVOLATILE RESIDUE Sample: 40 mL Analysis: Evaporate the Sample in a tared dish on a water bath, and dry at 105 for 1 h.
= area of the benzene peak from Sample solution A = area of the benzene peak from Standard solution D If necessary, the identity of benzene can be confirmed using another suitable chromatographic system (stationary phase with a different polarity). Acceptance criteria 3: NMT 2 ppm General acceptance criteria: The sum of all other impurities from Sample solution B is NMT the area of the peak due to 4-methylpentan-2-ol from Sample solution B (300 ppm). [NOTEDisregard any peaks that are 0.03 times the area of the peak corresponding to 4-methylpentan-2-ol fromSample solution B (9 ppm).] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. USP REFERENCE STANDARDS 11 USP Dehydrated Alcohol RS
BE BT
Dehydrated Alcohol Injection

(Comment on this Monograph)id=m1250=Dehydrated Alcohol Injection=A-Monos.pdf) DEFINITION Dehydrated Alcohol Injection is Dehydrated Alcohol suitable for parenteral use. IDENTIFICATION A. PROCEDURE Sample: Injection Analysis: Mix 5 drops of Sample in a small beaker with 1 mL of 1 /mL potassium permanganate and 5 drops of 2 N sulfuric acid, and cover the beaker immediately with a filter paper moistened with a solution recently prepared by dissolving 0.1 g of sodium nitroferricyanide and 0.25 g of piperazine in 5 mL of water. Acceptance criteria: An intense blue color is produced on the filter paper, the color becoming paler after a few minutes. B. PROCEDURE Sample solution: 0.1 mL/mL of Injection in water. Analysis: To 5 mL of the Sample solution, add 1 mL of 1.0 N sodium hydroxide, then slowly (over a period of 3 min) add 2 mL of 0.1 N iodine. Acceptance criteria: The odor of iodoform develops, and a yellow precipitate is formed within 30 min.
USP 32
Acceptance criteria: The weight of the residue does not exceed 1 mg. WATER-INSOLUBLE SUBSTANCES Sample: Injection Analysis: Dilute a portion of Sample with an equal volume of water. Acceptance criteria: The mixture is clear and remains clear for 30 min after cooling to 10. ULTRAVIOLET ABSORBANCE (See Spectroscopy and Light-Scattering 851.) Analytical wavelength: 235340 nm Cell: 1 cm Blank: Water Acceptance criteria: Absorbance is NMT 0.08 at 240 nm; and NMT 0.02 between 270 and 340 nm. [NOTEThe curve through these points is smooth.] OTHER REQUIREMENTS: Meets the requirements for Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose containers, preferably of Type I glass, and store at controlled room temperature. The container may contain an inert gas in the headspace.

Mode: UV Analytical Wavelength: 410 nm Cell: 1 cm Analysis: Samples: Standard solution, Sample solution, and Solution A. Transfer 10.0 mL each of the samples to individual 250-mL separators, add 40 mL of Solution A, 10 mL of Solution B, and 60 mL of chloroform. Shake the separators vigorously for 2 min, allow to stand for 15 min, then withdraw the chloroform layers through chloroform-washed cotton into 100-mL volumetric flasks. Repeat the extraction with 20 mL of chloroform, adding the filtered chloroform extracts to the respective volumetric flasks, and dilute with chloroform to volume. Without delay, concomitantly determine the absorbances of the solutions. [NOTEUse the solution prepared from Solution A as the Blank.] Calculate the quantity, in mg, of denatonium benzoate (C28H34N2O3 H2O) in 100 mL of Rubbing Alcohol taken: Result = (AU/AS) CS D = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Denatonium Benzoate RS in the Standard solution (mg/mL) D = sample dilution factor, 25 Acceptance criteria: NLT 1.40 mg of denatonium benzoate/100 mL of Rubbing Alcohol SUCROSE OCTAACETATE Sample solution: Using 50 mL of 70% alcohol, transfer the residue obtained in the test for Limit of Nonvolatile Residue to a 500-mL conical flask. Analysis: Neutralize the Sample solution with 0.1 N sodium hydroxide VS, using phenolphthalein TS as the indicator. Add 25.0 mL of 0.1 N sodium hydroxide, attach an air condenser to the flask, and reflux on a steam bath for 1 h. Remove from the steam bath, cool quickly, and titrate the excess alkali with 0.1 N sulfuric acid VS, using phenolphthalein TS as the indicator. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.1 N sodium hydroxide is equivalent to 8.483 mg of sucrose octaacetate (C28H38O19). Acceptance criteria: NLT 335 mg of sucrose octaacetate/100 mL of Rubbing Alcohol AU AS CS IMPURITIES Organic Impurities PROCEDURE: METHANOL Sample solution: 0.50 mL of sample diluted to 1.0 mL Analysis: To 0.50 mL of the Sample solution add 1 drop of dilute phosphoric acid (1 in 20) and 1 drop of 50 mg/mL potassium permanganate solution. Mix, allow to stand for 1 min, and add 50 mg/mL of sodium metabisulfite solution, dropwise, until the permanganate color is discharged. If a brown color remains, add 1 drop of dilute phosphoric acid (1 in 20). To the colorless solution add 5 mL of freshly prepared chromotropic acid TS, and heat in a water bath at 60 for 10 min. Acceptance criteria: No violet color appears. SPECIFIC TESTS SPECIFIC GRAVITY 841 Acceptance criteria: 0.86910.8771 at 15.56 (the U.S. Government standard temperature for alcohol determination) for Rubbing Alcohol manufactured with specially denatured alcohol Formula 23-H
Rubbing Alcohol
(Comment on this Monograph)id=m1270=Rubbing Alcohol=AMonos.pdf) DEFINITION Rubbing Alcohol and all preparations under the classification of Rubbing Alcohols are manufactured in accordance with the requirements of the U.S. Treasury Department, Bureau of Alcohol, Tobacco, and Firearms, Formula 23-H (8 parts by volume of acetone, 1.5 parts by volume of methyl isobutyl ketone, and 100 parts by volume of ethyl alcohol) being used. It contains NLT 68.5% and NMT 71.5% by volume of dehydrated alcohol, the remainder consisting of water and the denaturants, with or without color additives, and perfume oils. Rubbing Alcohol contains, in each 100 mL, NLT 355 mg of sucrose octaacetate or NLT 1.40 mg of denatonium benzoate. The preparation may be colored with one or more color additives, listed by the FDA for use in drugs. A suitable stabilizer may be added. Rubbing Alcohol complies with the requirements of the U.S. Treasury Department, Bureau of Alcohol, Tobacco, and Firearms. [NOTERubbing Alcohol is packaged, labeled, and sold in accordance with the regulations issued by the U.S. Treasury Department, Bureau of Alcohol, Tobacco, and Firearms.] ASSAY PROCEDURE [NOTEA sample must meet either the requirement for the Assay for Denatonium Benzoate or the requirement for the Assay for Sucrose Octaacetate.] DENATONIUM BENZOATE Solution A: 9.23 mg/mL of anhydrous dibasic sodium phosphate in water. Adjust with saturated citric acid solution to a pH of 4 0.1 before final dilution. Solution B: Bromophenol blue in chloroform (1 in 1000) Standard solution: 50 g/mL of USP Denatonium Benzoate RS Sample solution: Dissolve the residue obtained in the test for Limit of Nonvolatile Residue in 50.0 mL of water. Spectrometric conditions (See Spectoscopy and Light-Scattering 851.)
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A = length of the polarimeter tube (mm) R = observed rotation (degrees) Acceptance criteria: 95.0%105.0%
USP 32
LIMIT OF NONVOLATILE RESIDUE Where the denaturant is denatonium benzoate Sample: 200.0 mL Analysis: Evaporate the Sample, transferred in convenient portions, in a suitable tared dish on a steam bath, and dry the residue at 105 for 1 h. Acceptance criteria: The weight of the residue is not less than 2.8 mg. (Retain the residue for the Assay for Denatonium Benzoate.) Where the denaturant is sucrose octaacetate Sample: 25.0 mL Analysis: Evaporate the Sample in a suitable tared dish on a steam bath, and dry the residue at 105 for 1 h. Acceptance criteria: The weight of the residue is NLT 89 mg. (Retain the residue for the Assay for Sucrose Octaacetate.) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, remote from fire, and store at controlled room temperature. LABELING: Label it to indicate that it is flammable. USP REFERENCE STANDARDS 11 USP Denatonium Benzoate RS
IMPURITIES Inorganic Impurities HEAVY METALS 231 Sample: Equivalent to 4.0 g of dextrose adjusted to 25 mL by evaporation or by addition of water, as necessary. Acceptance criteria: NMT 5 C ppm, where C is the labeled amount, in g, of C6H12O6 H2O per mL of Injection Organic Impurities PROCEDURE: LIMIT OF 5-HYDROXYMETHYLFURFURAL AND RELATED SUBSTANCES Sample solution: 2 mg/mL of dextrose Spectrometric conditions (See Spectroscopy and Light-scattering 851.) Mode: UV Analytical wavelength: 284 nm Cell: 1 cm Blank: Water Analysis: Sample: Sample solution Acceptance criteria: The absorbance is NMT 0.25. SPECIFIC TESTS PH 791: 3.56.5 Sample solution: A portion of Dextrose injection to which 0.30 mL of saturated potassium chloride solution has been added for each 100 mL and which previously has been diluted with water, if necessary, to a concentration of NMT 5% of dextrose BACTERIAL ENDOTOXINS TEST 85: NMT 0.5 USP Endotoxin unit/mL OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose containers, preferably of Type I or Type II glass, and store at a controlled room temperature. LABELING: The label states the total osmolarity of the solution expressed in mOsmol/L. USP REFERENCE STANDARDS 11 USP Dehydrated Alcohol RS USP Endotoxin RS
Alcohol in Dextrose Injection

(Comment on this Monograph)id=m1320=Alcohol in Dextrose Injection=A-Monos.pdf) DEFINITION Alcohol in Dextrose Injection is a sterile solution of Alcohol and Dextrose in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of alcohol (C2H5OH), and NLT 95.0% and NMT 105.0% of the labeled amount of dextrose (C6H12O6 H2O). IDENTIFICATION A. PROCEDURE Sample solution: 50 L/mL of Dextrose Injection in water Analysis: Add a few drops of the Sample solution to 5 mL of hot alkaline cupric tartrate TS. Acceptance criteria: A copious red precipitate of cuprous oxide is formed. B. SPECIFIC GRAVITY 841: NMT 0.7962 at 15.56, indicating NLT 99.2% of C2H5OH by weight C. INFRARED ABSORPTION 197S or 197F: Meets the requirements ASSAY ALCOHOL DETERMINATION, Method 1Distillation Method 611 Sample: 50.0 mL Acceptance criteria: 90.0%110.0% DEXTROSE Sample: Amount equivalent to 25 g of dextrose Analysis: Transfer the Sample to a 100-mL volumetric flask. Add 0.2 mL of 6 N ammonium hydroxide, dilute with water to volume, and mix. Determine the angular rotation in a suitable polarimeter tube (see Optical Rotation 781). Calculate the percentage (g/100 mL) of C6H12O6 H2O taken: Result = [(Mr1/Mr2)/Rmid] 100/A R 100 Mr1 Mr2 Rmid = molecular weight of dextrose monohydrate, 198.17 = molecular weight of anhydrous dextrose, 180.16 = midpoint of the specific rotation range for anhydrous dextrose (degrees), 52.9
Alendronate Sodium
(Comment on this Monograph)id=m1324=Alendronate Sodium=A-Monos.pdf)
325.12 C4H12NNaO7P2 3H2O Phosphonic acid, (4-amino-1-hydroxybutylidene) bis-, monosodium salt, trihydrate; Sodium trihydrogen (4-amino-1hydroxybutylidene)diphosphonate, trihydrate [121268-17-5]. DEFINITION Alendronate Sodium contains NLT 98.0% and NMT 102.0% of C4H12NNaO7P2, calculated on the dried basis.
USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197M B. IDENTIFICATION TESTSGENERAL, Sodium 191: requirements of the flame test
Official Monographs / Alendronate 77

Acceptance criteria: Meets the 98.0%102.0%
ASSAY PROCEDURE Solution A: 14.7 mg/mL of sodium citrate dihydrate and 7.05 mg/mL of anhydrous dibasic sodium phosphate [NOTEAdjust solution to a pH of 8 with phosphoric acid.] Solution B: 19.1 mg/mL of sodium borate Solution C: 0.5 mg/mL of 9-fluorenylmethyl chloroformate in acetonitrile [NOTEPrepare this solution fresh just before use.] Mobile phase: Acetonitrile, methanol, and Solution A (25:5:70) Diluent: 29.4 mg/mL of sodium citrate dihydrate Standard stock solution: 0.1 mg/mL of USP Alendronate Sodium RS in Diluent Standard solution: Transfer 5.0 mL of the Standard stock solution to a 50-mL polypropylene, screw-cap centrifuge tube containing 5 mL of Solution B. Add 5 mL of Solution C, and shake for 30 s. Allow to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and shake vigorously for 1 min. Centrifuge for 510 min. Use a portion of the clear upper aqueous layer. Reagent blank: Transfer 5.0 mL of Diluent to a 50-mL polypropylene, screw-cap centrifuge tube containing 5 mL of Solution B. Add 5 mL of Solution C, and shake for 30 s. Allow to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and shake vigorously for 1 min. Centrifuge for 510 min. Use a portion of the clear upper aqueous layer. Sample stock solution: 0.1 mg/mL of Alendronate Sodium in Diluent Sample solution: Transfer 5.0 mL of the Sample stock solution to a 50-mL polypropylene, screw-cap centrifuge tube containing 5 mL of Solution B. Add 5 mL of Solution C, and shake for 30 s. Allow to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and shake vigorously for 1 min. Centrifuge for 510 min. Use a portion of the clear upper aqueous layer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 266 nm Column: 4.1-mm 25-cm; packing L21 Column temperature: 35 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution, Sample solution, and Reagent blank Calculate the percentage of C4H12NNaO7P2 in the portion of Alendronate Sodium taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak area from the Sample solution = peak area from the Standard solution = concentration of anhydrous sodium alendronate in the Standard stock solution (mg/mL) = concentration of the Sample stock solution (mg/mL)
IMPURITIES Inorganic Impurities HEAVY METALS, Method III 231: NMT 10 ppm Organic Impurities PROCEDURE Diluent: Prepare as directed in the Assay. Solution A: 2.94 mg/mL of sodium citrate dihydrate and 1.42 mg/mL of anhydrous dibasic sodium phosphate [NOTEAdjust solution to a pH of 8 with phosphoric acid, and then pass through a filter having a 0.5-m or finer porosity.] Solution B: Prepare as directed in the Assay. Solution C: 4 mg/mL of 9-fluorenylmethyl chloroformate in acetonitrile [NOTEPrepare this solution fresh just before use.] Solution D: Acetonitrile and Solution A (3:17) Solution E: Acetonitrile and Solution A (7:3) Mobile phase: See the gradient table below.
Time (min) 0 15 25 27 32 Solution D (%) 100 50 0 100 100 Solution E (%) 0 50 100 0 0
Standard stock solution: 0.6 mg/mL of USP Alendronate Sodium RS in Diluent Standard solution A: Transfer 5.0 mL of the Standard stock solution to a 50-mL polypropylene, screw-cap centrifuge tube containing 5 mL of Solution B. Add 5 mL of acetonitrile and 5 mL of Solution C, and shake for 45 s. Allow to stand at room temperature for 30 min. Add 20 mL of methylene chloride, and shake vigorously for 1 min. Centrifuge for 5 to 10 min, and use a portion of the clear upper aqueous layer. Standard solution B: Dilute the Standard stock solution to 0.6 g/mL with Diluent. Transfer 5 mL to a 50-mL polypropylene, screw-cap centrifuge tube containing 5 mL of Solution B. Add 5 mL of acetonitrile and 5 mL of Solution C, and shake for 45 s. Allow to stand at room temperature for 30 min. Add 20 mL of methylene chloride, and shake vigorously for 1 min. Centrifuge for 5 to 10 min, and use a portion of the clear upper aqueous layer. Blank: Transfer 5.0-mL of Diluent to a 50-mL polypropylene, screw-cap centrifuge tube containing 5 mL of Solution B. Add 5 mL of acetonitrile and 5 mL of Solution C, and shake for 45 s. Allow to stand at room temperature for 30 min. Add 20 mL of methylene chloride, and shake vigorously for 1 min. Centrifuge for 5 to 10 min, and use a portion of the clear upper aqueous layer. Sample stock solution: 0.6 mg/mL of Alendronate Sodium in Diluent Sample solution: Transfer 5.0-mL of Sample stock solution to a 50-mL polypropylene, screw-cap centrifuge tube containing 5 mL of Solution B. Add 5 mL of acetonitrile and 5 mL of Solution C, and shake for 45 s. Allow to stand at room temperature for 30 min. Add 20 mL of methylene chloride, and shake vigorously for 1 min. Centrifuge for 5 to 10 min, and use a portion of the clear upper aqueous layer. Chromatographic system (See Chromatography 621, System Suitability.)
78
Alendronate / Official Monographs

Mode: LC Detector: UV 266 nm Column: 4.1-mm 25-cm; packing L21 Column temperature: 45 Flow rate: 1.8 mL/min Injection size: 20 L System suitability Samples: Standard solution A and Standard solution B Suitability requirements Tailing factor: NMT 2.0 for the main peak, Standard solution A Signal-to-noise ratio: NLT 3 for Standard solution B for the peak at the locus corresponding to the main peak of Standard solution A Analysis Samples: Sample solution and Blank [NOTEDisregard any peak corresponding to those obtained from the Blank.] Calculate the percentage of each impurity in the portion of Alendronate Sodium taken: Result = (rU/rT) 100 = area of each impurity peak rU = sum of all impurity peaks and the main peak rT Acceptance criteria Individual impurities: NMT 0.1% Total impurities: NMT 0.5%
USP 32
Diluent: 29.4 mg/mL of sodium citrate dihydrate Standard stock solution: 0.03 mg/mL of anhydrous USP Alendronate Sodium RS in Diluent Standard solution: Transfer 5.0 mL of the Standard stock solution to a 50-mL polypropylene screw-cap centrifuge tube containing 5 mL of Solution B, and mix for 3 min. Add 4 mL of Solution C, and agitate for 30 s. Allow the solution to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and agitate for 40 s. Centrifuge the mixture for 10 min. Use the clear upper aqueous layer. Sample stock solution: Transfer NLT 10 Tablets to a 1000mL volumetric flask. Add 500 mL of Diluent, shake by mechanical means for 30 min, and sonicate for 5 min. Dilute with Diluent to volume, and centrifuge a portion of this solution. Quantitatively dilute a portion of the clear supernatant to a concentration equivalent to 0.020.03 mg/mL of alendronate sodium. Sample solution: Transfer 5.0 mL of the Sample stock solution to a 50-mL polypropylene screw-cap centrifuge tube containing 5 mL of Solution B, and mix for 3 min. Add 4 mL of Solution C, and agitate for 30 s. Allow the solution to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and agitate for 40 s. Centrifuge the mixture for 10 min. Use the clear upper aqueous layer. Blank: Transfer 5 mL of Diluent to a 50-mL polypropylene screw-cap centrifuge tube containing 5 mL of Solution B, and mix for 3 min. Add 4 mL of Solution C, and agitate for 30 s. Allow the solution to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and agitate for 40 s. Centrifuge the mixture for 10 min. Use the clear upper aqueous layer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 266 nm Column: 4.1-mm 25-cm; packing L21 Column temperature: 35 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 2.0 Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution, Sample solution, and Blank Calculate the percentage of the label claim in the portion of C4H13NO7P2 taken: Result = (rU/rS) (CS/CU) F 100/L = peak area from the Sample solution = peak area from the Standard solution = concentration of anhydrous USP Alendronate Sodium RS in the Standard stock solution (mg/mL) = concentration of the Tablet in the Sample stock CU solution (Tablets/mL) F = molecular weight conversion factor (C4H13NO7P2/C4H12NNaO7P2), 0.919 L = label claim (mg/Tablet) Acceptance criteria: C4H12NNaO7P2 equivalent to 90.0%110.0% of the labeled amount of C4H13NO7P2 rU rS CS
SPECIFIC TESTS LOSS ON DRYING 731: Dry it at a pressure of NMT 5 mm of mercury at 140 to constant weight: it loses NLT 16.1% and NMT 17.1% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store at room temperature. USP REFERENCE STANDARDS 11 USP Alendronate Sodium RS
Alendronate Sodium Tablets

(Comment on this Monograph)id=m1327=Alendronate Sodium Tablets=A-Monos.pdf) DEFINITION Alendronate Sodium Tablets contain an amount of Alendronate Sodium equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of alendronic acid (C4H13NO7P2). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 14.7 mg/mL of sodium citrate dihydrate and 7.05 mg/mL of anhydrous dibasic sodium phosphate [NOTEAdjust pH to 8.0 with phosphoric acid before bringing the solution to volume.] Solution B: 38.1 mg/mL of sodium borate Solution C: 1 mg/mL of 9-fluorenylmethyl chloroformate in acetonitrile [NOTEPrepare this solution fresh just before use.] Mobile phase: Acetonitrile, methanol, and Solution A (20:5:75)
USP 32
PERFORMANCE TESTS Change to read: DISSOLUTION 711 Test 13 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 15 min Determine the amount of C4H13NO7P2 dissolved by using the following method. Solution A and Mobile Phase: Proceed as directed in the Assay. Diluent: 176.4 mg/mL of sodium citrate in Medium Solution B: Dissolve 6.2 g of boric acid in approximately 950 mL of water. Adjust with 1 N sodium hydroxide to a pH of 9.0, and dilute with water to 1 L. Solution C: 0.5 mg/mL of 9-fluorenylmethyl chloroformate in acetonitrile [NOTEPrepare this solution fresh.] Standard stock solution: USP Alendronate Sodium RS in Medium to make a concentration equivalent to dissolving 1 Tablet of sample in 900 mL of the same Medium Standard solution: Transfer 5.0 mL of the Standard stock solution to a 50-mL polypropylene screw-cap centrifuge tube containing 1.0 mL of Diluent and 5.0 mL of Solution B, and mix for 3 min. Add 4.0 mL of Solution C, and agitate for 30 s. Allow the solution to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and agitate for 40 s. Centrifuge the mixture for 5 min. Use a portion of the clear, upper aqueous layer. Blank: Transfer 5 mL of water to a 50-mL polypropylene screw-cap centrifuge tube containing 1.0 mL of Diluent and 5.0 mL of Solution B, and mix for 3 min. Add 4.0 mL of Solution C, and agitate for 30 s. Allow the solution to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and agitate for 40 s. Centrifuge the mixture for 5 min. Use a portion of the clear, upper aqueous layer. Sample stock solution: Withdraw a portion of the solution under test, centrifuge immediately, and use a portion of the clear upper supernatant. Sample solution: Transfer 5.0 mL of the Sample stock solution to a 50-mL polypropylene screw-cap centrifuge tube containing 1.0 mL of Diluent and 5.0 mL of Solution B, and mix for 3 min. Add 4.0 mL of Solution C, and agitate for 30 s. Allow the solution to stand at room temperature for 25 min. Add 25 mL of methylene chloride, and agitate for 40 s. Centrifuge the mixture for 5 min. Use a portion of the clear, upper aqueous layer. Chromatographic system and System suitabilitity: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C4H13NO7P2 dissolved: Result = (rU/rS) CS F V 100/L = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Alendronate Sodium RS in the Standard solution (mg/mL) F = molecular weight conversion factor (C4H13NO7P2/C4H12NNaO7P2), 0.919 V = volume of the Medium, 900 mL L = Tablet label claim (mg) Tolerances: NLT 80% (Q) of the labeled amount of C4H13NO7P2 is dissolved; for tablets labeled for weekly dosing, NLT 75% (Q) of the labeled amount of C4H13NO7P2 is dissolved. rU rS CS
Test 2
Official Monographs / Alfentanil 79
If the product complies with this test, the labeling indicates that the product meets USP Dissolution Test 2. Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of C4H12NNaO7P2 3H2O dissolved as directed in the Assay. Tolerances: NLT 80% (Q) of the labeled amount of alendronate sodium (C4H12NNaO7P2 3H2O) is dissolved.3 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 15 and 30. Change to read: LABELING: The labeling indicates weekly dosing where appropriate. When more than one Dissolution test is given, the labeling states the test used only if Test 1 is not used.3 USP REFERENCE STANDARDS 11 USP Alendronate Sodium RS
Alfentanil Hydrochloride
(Comment on this Monograph)id=m1350=Alfentanil Hydrochloride=A-Monos.pdf)
C21H32N6O3 HCl H2O
470.99
452.98 C21H32N6O3 HCI Propanamide, N-[1-[2-(4-ethyl-4,5-dihydro-5-oxo-1H-tetrazol-1yl)ethyl]-4-(methoxymethyl)-4-piperidinyl]-N-phenyl, monohydrochloride, monohydrate; N-[1-[2-(4-Ethyl-5-oxo-2-tetrazolin-1-yl)-ethyl]-4(methoxymethyl)-4-piperidyl]propionanilide monohydrochloride monohydrate [70879-28-6]. Anhydrous [69049-06-5]. DEFINITION Alfentanil Hydrochloride contains NLT 98.0% and NMT 102.0% of C21H32N6O3 HCl, calculated on the anhydrous basis. [CAUTIONHandle Alfentanil Hydrochloride with great care because it is a potent opioid analgesic. Great care should be taken to prevent inhaling particles of Alfentanil Hydrochloride and exposing the skin to it.] IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample solution: 11.7 mg/mL of Alfentanil Hydrochloride in glacial acetic acid. Analysis: To 30 mL of Sample solution add 3 mL of mercuric acetate TS and 3 drops of p-naphtholbenzein TS, and titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 45.298 mg of C21H32N6O3 HCl.
80
Alfentanil / Official Monographs

98.0%102.0%
USP 32
Visualizing agent: Dragendorffs reagent Analysis Samples: Standard solution and Sample solution Proceed as directed in the chapter. Acceptance criteria: Meets the requirements B. The retention time of the major peak from the Sample solution corresponds to that from the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.01 M tetrabutylammonium hydrogen sulfate (86:14) Standard solution: 0.54 mg/mL of USP Alfentanil Hydrochloride RS in saline TS Sample solution: 0.50 mg/mL of Alfentanil from volume of Injection in saline TS Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; contains spherical 5-m packing L1 Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 5400 theoretical plates Tailing factor: NMT 1.3 Relative standard deviation: NMT 1.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H32N6O3 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Alfentanil Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of alfentanil in the Sample solution (mg/mL) = molecular weight for alfentanil, 416.52 Mr1 = molecular weight for alfentanil hydrochloride, Mr2 452.98 Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE Mobile phase: Acetonitrile and 0.01 M tetrabutylammonium hydrogen sulfate (86:14) Standard solution: 0.54 mg/mL of USP Alfentanil Hydrochloride RS in saline TS Sample solution: 0.50 mg/mL of Alfentanil from volume of Injection in saline TS Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; contains spherical 5-m packing L1 rU rS CS
Acceptance criteria:
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Mobile phase: Acetonitrile and 0.01 M tetrabutylammonium hydrogen sulfate (86:14) Standard solution: 0.54 mg/mL of USP Alfentanil Hydrochloride RS in Mobile phase Sample solution: 0.54 mg/mL of Alfentanil Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; contains spherical 5-m packing L1 Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 5400 theoretical plates Tailing factor: NMT 1.3 Relative standard deviation: NMT 1.0% for replicate injections Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Alfentanil Hydrochloride taken: Result = (rU/rT) 100 = response of each impurity peak rU = sum of all of the peaks rT Acceptance criteria Any single impurity: NMT 0.5% Total impurities: NMT 1.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 4.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Alfentanil Hydrochloride RS
Alfentanil Injection
(Comment on this Monograph)id=m1355=Alfentanil Injection=A-Monos.pdf) DEFINITION Alfentanil Injection is a sterile solution of Alfentanil Hydrochloride in Water for Injection. It contains an amount of Alfentanil Hydrochloride equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of C21H32N6O3. [CAUTIONHandle Alfentanil Injection with great care because it is a potent opioid analgesic.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 0.54 mg/mL of USP Alfentanil Hydrochloride RS Sample solution: 0.5 mg/mL of alfentanil in water Application volume: 200 L Developing solvent system: Chloroform, methanol, and formic acid (85:10:5)
USP 32
Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 5400 theoretical plates Tailing factor: NMT 1.3 Relative standard deviation: NMT 1.0% for replicate injections Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Injection taken: Result = (rU/rT) 100 rU = response of each impurity peak = sum of all of the peaks rT Acceptance criteria: The sum of all impurities is NMT 2.0%. SPECIFIC TESTS PH 791: 4.06.0 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections BACTERIAL ENDOTOXINS TEST 85: NMT 10 USP Endotoxin Units/mL OTHER REQUIREMENTS: Meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose or multiple-dose containers, preferably of Type I glass, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Alfentanil Hydrochloride RS USP Endotoxin RS
Official Monographs / Alfuzosin 81

acid. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 M perchloric acid titrant is equivalent to 42.59 mg of C19H27N5O4 HCl. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Solution A: 58.5 mM perchloric acid. Adjust with 2 M sodium hydroxide to a pH of 3.5 before final dilution. Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A (20:1:80) System suitability solution: 0.4 mg /mL of USP Alfuzosin Hydrochloride System Suitability Mixture RS in Mobile phase Sample solution A: 0.40 mg/mL of Alfuzosin Hydrochloride in Mobile phase Sample solution B: 0.40 g/mL of Alfuzosin Hydrochloride, from Sample solution A, in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5 m packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Peak-to-valley ratio: NLT 5 for impurity A and alfuzosin Calculate the peak-to-valley ratio: Result = (HP/HV) HP HV = height of impurity A peak above baseline = lowest point between the impurity A peak and the alfuzosin peak.
Alfuzosin Hydrochloride
(Comment on this Monograph)id=m891=Alfuzosin Hydrochloride=A-Monos.pdf)
Analysis Samples: Sample solution A and Sample solution B Calculate the percentage of each impurity in the portion of Alfuzosin Hydrochloride taken: Result = (rU/rS) (CS/CU) 100
425.91 C19H27N5O4 HCl 2-Furancarboxamide, ()-N-[3-[(4-amino-6,7-dimethoxy-2quinazolinyl)methylamino]propyl]tetrahydro-, monohydrochloride; ()-N-[3-[(4-Amino-6,7-dimethoxy-2quinazolinyl)methylamino]propyl]tetrahydro-2-furamide monohydrochloride [81403-68-1]. DEFINITION Alfuzosin Hydrochloride contains NLT 99.0% and NMT 101.0% of C19H27N5O4 HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Chloride 191: requirements
= peak response of each impurity from Sample solution A = peak response from Sample solution B rS = concentration of Sample solution B (mg/mL) CS CU = concentration of the Sample solution A (mg/mL) Acceptance criteria Individual impurities: See Impurity Table 1. [NOTEDisregard any peaks less than 0.05%.] Total impurities: NMT 0.30 %
Impurity Table 1 Name Impurity Aa Impurity D
a
rU
Relative Retention Time 1.2 0.5
Acceptance Criteria NMT % b 0.20
Meets the
ASSAY PROCEDURE Sample: 300 mg Analysis: Dissolve the Sample in 80 mL of glacial acetic acid and acetic anhydride (1:1). Titrate with 0.1 M perchloric
N-[3-[(4-Amino-6,7-dimethoxyquinazolin-2yl)(methyl)amino]propyl]furan-2-carboxamide. b Impurity A, a component of USP Alfuzosin System Suitability Mixture RS, is not a specified impurity. c N-(4-Amino-6,7-dimethoxyquinazolin-2-yl)-N-methylpropane-1,3-diamine.
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Impurity Table 1 (continued) Name Relative Retention Time 1.0 Acceptance Criteria NMT % 0.10
USP 32
a suitable electrode system (see Titrimetry 541). Each mL of 0.1 M sodium hydroxide is equivalent to 15.81 mg of C4H6N4O3. Acceptance criteria: NLT 98.5% and NMT 101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Adsorbent: Cellulose Standard solution A: 1 mg/mL of USP Allantoin RS in methanol and water (1:1) Urea stock solution: 1 mg/mL of USP Urea RS Standard solution B: 0.1 mg/mL in methanol, from Urea stock solution Standard solution C: Standard solution A and Standard solution B (1:1) Sample solution A: Transfer 0.10 g of Allantoin to a 10-mL volumetric flask, add 5 mL of water, dissolve by heating, and allow to cool. Dilute with methanol to volume. [NOTEUse immediately after preparation.] Sample solution B: Transfer 1 mL of Sample solution A to a 10-mL volumetric flask, and dilute with a mixture of methanol and water (1:1) to volume. Spray reagent: 5 mg/mL of pdimethylaminobenzaldehyde in a mixture of methanol and hydrochloric acid (3:1) Application volume: 5 or 10 L for Sample solution A Developing solvent system: Butyl alcohol, glacial acetic acid, and water (60:15:25) Analysis Samples: Standard solution A, Standard solution B, Standard solution C, Sample solution A, and Sample solution B Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram until the solvent front has moved about 10 cm. Spray the plate with Spray reagent, dry in a current of hot air, and after 30 min examine under visible light. Acceptance criteria: Any spot from Sample solution A, except for the principal spot, is not more intense than the spot from Standard solution B: NMT 0.5% of any individual impurity. [NOTEThe test is not valid unless the principal spots from Standard solution C are clearly separated.] SPECIFIC TESTS OPTICAL ROTATION, Angular Rotation 781A Sample solution: 10 mg/mL, in carbon dioxide-free water [NOTEReserve a portion of this solution for use in the test for Acidity.] Acceptance criteria: 10 to +10 ACIDITY OR ALKALINITY Sample: Use the Sample solution retained from the test for Angular Rotation. Analysis: To 5 mL of the Sample add 5 mL of water, 0.1 mL of methyl red TS, and 0.2 mL of 0.01 M sodium hydroxide. Acceptance criteria: A yellow color is observed. Solution turns red upon addition of 0.4 mL of 0.01 M hydrochloric acid. LOSS ON DRYING 731: Dry a sample at 105 to constant weight: it loses NMT 0.1% of its weight. REDUCING SUBSTANCES Sample solution: 1.0 g of Allantoin in 10 mL of water, shaken for 2 min, and filtered Analysis: To the Sample solution add 1.5 mL of 0.02 M potassium permanganate. Acceptance criteria: The solution remains violet for at least 10 min.
Alfuzosin Any other individual, unidentified impurity

a
N-[3-[(4-Amino-6,7-dimethoxyquinazolin-2yl)(methyl)amino]propyl]furan-2-carboxamide. b Impurity A, a component of USP Alfuzosin System Suitability Mixture RS, is not a specified impurity. c N-(4-Amino-6,7-dimethoxyquinazolin-2-yl)-N-methylpropane-1,3-diamine.
SPECIFIC TESTS OPTICAL ROTATION 781: 10 to +10 Sample solution: 20 mg/mL in carbon dioxide-free water WATER DETERMINATION, Method I 921: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Protect from light and moisture, and store at room temperature. USP REFERENCE STANDARDS 11 USP Alfuzosin Hydrochloride RS USP Alfuzosin System Suitability Mixture RS
Allantoin
(Comment on this Monograph)id=m1400=Allantoin=AMonos.pdf)
C4H6N4O3 Urea, (2,5-dioxo-4-imidazolidinyl)- ; Allantoin [97-59-6].
158.12
DEFINITION Allantoin contains NLT 98.5% and NMT 101.0% of C4H6N4O3. IDENTIFICATION A. INFRARED ABSORPTION 197K B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201: The RF value of the principal spot from Sample solution A corresponds to that from Standard solution A, as described in Organic Impurities. C. PROCEDURE Sample solution: Dissolve 20 mg of Allantoin in 2 mL of 1 M sodium hydroxide. Heat to boiling, allow to cool, and add 1 mL of 2 M hydrochloric acid. Analysis: To 0.1 mL of Sample solution add 0.1 mL of 100 mg/mL of potassium bromide solution, 0.1 mL of 20 mg/mL of resorcinol solution, and 3 mL of sulfuric acid. Heat on a water bath for 510 min. Acceptance criteria: A dark blue color, which turns red after cooling and pouring into about 10 mL of water, is observed. ASSAY Procedure Sample: 120 mg Analysis: Transfer the Sample to a 100-mL beaker, dissolve by stirring in 40 mL of water, and titrate with 0.1 M sodium hydroxide. Determine the endpoint potentiometrically, using
USP 32
ADDITIONAL REQUIREMENTS USP REFERENCE STANDARDS 11 USP Allantoin RS USP Urea RS
Official Monographs / Allopurinol 83

Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.8 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for allopurinol related compound B, for allopurinol related compound C, and for allopurinol are about 0.7, 0.8, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.1 between allopurinol related compound B and allopurinol related compound C and NLT 6.0 between allopurinol related compound C and allopurinol, System suitability solution Relative standard deviation: NMT 2.0% for replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C5H4N4O in the portion of Allopurinol taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Allopurinol RS in Standard solution (mg/mL) CU = concentration of Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Change to read: Organic Impurities PROCEDURE 1 [NOTEStore and inject the Standard solution and the Sample solution at 8, using a cooled autosampler.] Solution A: 1.25 mg/mL of monobasic potassium phosphate in water Solution B: Methanol Diluent: Solution A and Solution B (9:1) vAllopurinol related compound F solution: Weigh 5 mg of USP Allopurinol Related Compound F RS into a suitable flask. Add 2.0 mL of 0.1 N sodium hydroxide, promptly sonicate with swirling for 13 min to dissolve, add 80 mL of Diluent, and sonicate for an additional 5 min.vUSP32 Standard stock solution: Transfer 5 mg of each of USP Allopurinol RS, USP Allopurinol Related Compound A RS, USP Allopurinol Related Compound B RS, USP Allopurinol Related Compound C RS, USP Allopurinol Related Compound D RS, vandvUSP32 USP Allopurinol Related Compound E RS vvUSP32 to a 100-mL volumetric flask. Add 2.0 mL of 0.1 N sodium hydroxide to dissolve, vadd the entire volume of Allopurinol related compound F solution to the flask, rinse the flask in which Allopurinol related compound F solution that was prepared with a small amount of Diluent, and add the rinsings to the solutionvUSP32. Sonicate for an additional 5 min. Dilute with Diluent to volume. [NOTEThis solution is stable for 48 h when stored at 8.] Standard solution: 0.5 g/mL of allopurinol, and allopurinol related compounds A, B C, D, E, and F from the Standard stock solution in Diluent.
Allopurinol
(Comment on this Monograph)id=m1430=Allopurinol=AMonos.pdf)
C5H4N4O 4H-Pyrazolo[3,4-d]pyrimidin-4-one, 1,5-dihydro-; 1,5-Dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one; 1H-Pyrazolo[3,4-d]pyrimidin-4-ol. [315-30-0].
136.11
DEFINITION Allopurinol contains NLT 98.0% and NMT 102.0% of C5H4N4O, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE [NOTEStore and inject the System suitability solution, the Standard solution, and the Sample solution at 8, using a cooled autosampler.] Mobile phase: 1.25 mg/mL of monobasic potassium phosphate System suitability stock solution 1: 50 g/mL of USP Allopurinol RS in Mobile phase. [NOTEDissolve the reference standard in a small amount of 0.1 N sodium hydroxide before diluting with Mobile phase.] System suitability stock solution 2: 50 g/mL of USP Allopurinol Related Compound B RS in Mobile phase. [NOTEDissolve the reference standard in a small amount of 0.1 N sodium hydroxide before diluting with Mobile phase.] System suitability stock solution 3: 50 g/mL of USP Allopurinol Related Compound C RS in Mobile phase. [NOTEDissolve the reference standard in a small amount of 0.1 N sodium hydroxide before diluting with Mobile phase.] System suitability solution: Transfer 1.0 mL of each of the three System suitability stock solutions to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Standard stock solution: 0.5 mg/mL of USP Allopurinol RS, prepared by dissolving the USP Allopurinol RS in a small volume of 0.1 N sodium hydroxide and immediately diluting to the appropriate volume with Mobile phase. Standard solution: 80 g/mL of USP Allopurinol RS from the Standard stock solution in Mobile phase. Sample stock solution: Dissolve 50 mg of Allopurinol in 5.0 mL of 0.1 N sodium hydroxide in a 100-mL volumetric flask. Immediate dilute with Mobile phase to volume. Sample solution: 80 g/mL of Allopurinol from the Sample stock solution in Mobile phase. Chromatographic system (See Chromatography 621, System Suitability.)
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Sample solution: Transfer 25 mg of Allopurinol to a 100mL volumetric flask. Add 5.0 mL of 0.1 N sodium hydroxide to dissolve, promptly sonicate with swirling for NMT 1 min, add 80 mL of Diluent, and sonicate for an additional 5 min. Dilute with Diluent to volume. Mobile phase: Use gradient table below. below.
Time (min) 0 30 35 36 46 Solution A (%) 90 70 70 90 90 Solution B (%) 10 30 30 10 10 Impurity Table 1 (continued) Relative Retention Time 0.81 1.0 4.4 4.8 6.5 Relative Response Factor
USP 32
Name Allopurinol related compound Bb Allopurinol Allopurinol related compound Dd Allopurinol related compound Ee Allopurinol related compound Ff
a b
Acceptance Criteria NMT % 0.2 0.2 0.2 0.2
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Column temperature: 30 Flow rate: 1 mL/min Injection size: 40 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 0.8 between allopurinol related compounds C and B Tailing factor: NMT 1.5 for the allopurinol peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Allopurinol taken: Result = (rU/rS) (CS/CU) 100 = peak response for each individual impurity from the Sample solution = peak response for each individual impurity from rS the Standard solution [NOTEFor unspecified impurities, rS is the peak response for the allopurinol peak from the Standard solution.] = concentration of each individual impurity in the CS Standard solution (mg/mL) = concentration of Allopurinol in the Sample CU solution (mg/mL) Acceptance criteria Individual specified impurities: see Impurity Table I. Individual unspecified impurities: NMT 0.1% Total impurities: NMT 1.0%
Impurity Table 1 Relative Retention Time 0.62 0.79 Relative Response Factor Acceptance Criteria NMT % 0.2 0.2
3-Amino-1H-pyrazole-4-carboxamide. 5-(Formylamino)-1H-pyrazole-4-carboxamide. c 5-(4H-1,2,4-Triazol-4-yl)-1H-pyrazole-4-carboxamide. d Ethyl-5-amino-1H-pyrazole-4-carboxylate. e Ethyl-5-(formylamino)-1H-pyrazole-4-carboxylate. f Ethyl-(E/Z)-3-(2-carbethoxy-2-cyanoethenyl)amino-1H-pyrazole-4carboxylate.
rU
Name Allopurinol related compound Aa Allopurinol related compound Cc

a b
3-Amino-1H-pyrazole-4-carboxamide. 5-(Formylamino)-1H-pyrazole-4-carboxamide. c 5-(4H-1,2,4-Triazol-4-yl)-1H-pyrazole-4-carboxamide. d Ethyl-5-amino-1H-pyrazole-4-carboxylate. e Ethyl-5-(formylamino)-1H-pyrazole-4-carboxylate. f Ethyl-(E/Z)-3-(2-carbethoxy-2-cyanoethenyl)amino-1H-pyrazole-4carboxylate.
Procedure 2: Limit of Hydrazine [NOTEUnder the following conditions, any hydrazine present in the sample will react with benzaldehyde to form benzalazine.] Mobile phase: Hexane and isopropyl alcohol (95:5) Solution A: 85 mg/mL sodium hydroxide [NOTE Alternatively, a commercially available 2 N sodium hydroxide solution can be used.] Diluent: Methanol and Solution A (1:1) Solution B: 40 mg/mL benzaldehyde in Diluent Standard stock solution: 2.0 g/mL hydrazine sulfate in Diluent [NOTESonicate, as necessary, to facilitate dissolution.] Standard solution: Mix 5.0 mL of the Standard stock solution and 4 mL of Solution B. Allow to stand for 2.5 h at room temperature. Add 5.0 mL of hexane, and shake for 1 min. Allow the layers to separate, and use the upper (hexane) layer. Sample stock solution: Dissolve 250.0 mg of Allopurinol in 5.0 mL of Diluent. Sample solution: To the entire volume of Sample stock solution add 4 mL of Solution B, mix, and allow to stand for 2.5 h at room temperature. Add 5.0 mL of hexane, and shake for 1 min. Allow the layers to separate, and use the upper (hexane) layer. Blank solution: Mix 5.0 mL of Diluent and 4 mL of Solution B, and allow to stand for 2.5 h at room temperature. Add 5.0 mL of hexane, and shake for 1 min. Allow the layers to separate, and use the upper (hexane) layer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 310 nm Column: 4.6-mm 25-cm; 5-m packing L10 Column temperature: 30 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for benzalazine and benzaldehyde are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between benzalazine and benzaldehyde Relative standard deviation: NMT 15.0% for the benzalazine peak
v
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the amount, in ppm, of hydrazine in the portion of Allopurinol taken: Result = (rU/rS) (CS/CT) (Mr1/Mr2) F = peak response for the benzalazine peak from the Sample solution rS = peak response for the benzalazine peak from the Standard solution = concentration of hydrazine sulfate in the CS Standard stock solution (g/mL) = concentration of allopurinol in the Sample stock CT solution (mg/mL) Mr1 = molecular weight of hydrazine, 32.05 Mr2 = molecular weight of hydrazine sulfate, 130.12 F = unit conversion factor (from g/mg to ppm), 1000 Acceptance criteria: NMT 10 ppm of hydrazinevUSP32 SPECIFIC TESTS LOSS ON DRYING 731: Dry a sample in vacuum at 105 for 5 h: it loses NLT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at room temperature. USP REFERENCE STANDARDS 11 USP Allopurinol RS USP Allopurinol Related Compound A RS USP Allopurinol Related Compound B RS USP Allopurinol Related Compound C RS USP Allopurinol Related Compound D RS USP Allopurinol Related Compound E RS USP Allopurinol Related Compound F RS rU
Official Monographs / Allopurinol 85

LABELING: Label it to state that it is to be shaken well before use and that it is to be discarded after 60 days. Label it to state that it is to be kept out of the reach of children. Label it to indicate the nominal content of allopurinol in the Oral Suspension. BEYOND-USE DATE: NMT 60 days after preparation
Allopurinol Tablets
(Comment on this Monograph)id=m1460=Allopurinol Tablets=A-Monos.pdf) DEFINITION Allopurinol Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of C5H4N4O. IDENTIFICATION INFRARED ABSORPTION 197K Sample: Extract a quantity of finely powdered Tablets equivalent to 50 mg of allopurinol by trituration with 10 mL of 0.1 N sodium hydroxide. Filter, acidify the filtrate with 1 N acetic acid, collect the precipitated allopurinol (allow 1015 min for sufficient precipitation to occur), wash the precipitate with 3 mL of dehydrated alcohol, in portions, and finally wash with 4 mL of anhydrous ethyl ether. Allow to dry in air for 15 min, then dry at 105 for 3 h. ASSAY PROCEDURE [NOTEDo not allow the Mobile phase to remain in the column overnight. After performing the procedure, flush the system with water for NLT 20 min, and then flush with methanol for 20 min.] Mobile phase: 0.05 M monobasic ammonium phosphate Internal standard solution: Dissolve 50 mg of hypoxanthine in 10 mL of 0.1 N sodium hydroxide, shake by mechanical means until dissolved (about 10 min), and dilute with water to 50 mL. [NOTEPrepare on the day of use.] Standard stock solution: Transfer 50 mg of USP Allopurinol RS to a 50-mL volumetric flask. Add 10 mL of 0.1 N sodium hydroxide, shake by mechanical means until dissolved (about 10 min), and dilute with water to volume. [NOTE Prepare on the day of use.] Standard solution: Transfer 4.0 mL of the Standard stock solution to a 200-mL volumetric flask. Add 2.0 mL of the Internal standard solution, and dilute with Mobile phase to volume. Sample stock solution: Transfer an amount of finely powdered Tablets (NLT 20 Tablets) equivalent to 50 mg of allopurinol to a 500-mL volumetric flask. Add 10 mL of 0.1 N sodium hydroxide, shake by mechanical means for 10 min, and add water to volume. [NOTEFrom this point, conduct the remainder of the Assay without delay.] Filter, rejecting the first 10 mL of the filtrate. Sample solution: Transfer 4.0 mL of the Sample stock solution to a 200-mL volumetric flask. Add 2.0 mL of Internal standard solution, and dilute with Mobile phase to volume. Chromtographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 15 L System suitability Sample: Standard solution [NOTEThe relative retention times for hypoxanthine and allopurinol are about 0.6 and 1.0, respectively.]
Allopurinol Oral Suspension

(Comment on this Monograph)id=m385=Allopurinol Oral Suspension=A-Monos.pdf) DEFINITION Allopurinol Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of allopurinol (C5H4N4O). Prepare Allopurinol Oral Suspension at a 2% concentration, for example, as follows (see Pharmaceutical Compounding Nonsterile Preparations 795).
Allopurinol Glycerin Vehicle for Oral Suspension, NF Vehicle for Oral Solution, NF To make 2g 5 mL 45 mL A sufficient quantity 100 mL
Select the number of Tablets that contain the specified amount of allopurinol and calculate the quantity of each ingredient required for the total amount to be prepared. Count/weigh/ measure each ingredient. Thoroughly pulverize the tablets. Mix the powdered Allopurinol Tablets and Glycerin to form a smooth paste, incorporate the Vehicle for Oral Suspension, add sufficient Vehicle for Oral Solution to volume, and mix well. Adjust the pH, if necessary. Package, and label. SPECIFIC TESTS PH 791: An apparent pH between 6.5 and 7.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Package in a tight container, and store at controlled room temperature.
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USP 32
DEFINITION Allyl Isothiocyanate contains NLT 93.0% and NMT 105.0% of C4H5NS. [CAUTIONAllyl Isothiocyanate is a potent lachrymator, with a pungent irritating odor. Care should be taken to protect the eyes, to prevent inhalation of fumes, and to avoid tasting.] IDENTIFICATION INFRARED ABSORPTION 197F: The spectrum exhibits pronounced peaks at about 700cm, 950cm, 980cm, 1300cm, 1340cm, 1350cm, 1410cm, 1420cm, 1650cm, 2100cm, and 2200cm. ASSAY PROCEDURE Sample solution: Dilute a 4-mL sample to 100 mL in a volumetric flask with alcohol. Analysis: Transfer 5.0 mL of Sample solution to a 100-mL conical flask. Add 50.0 mL of 0.1 N silver nitrate VS and 5 mL of ammonia TS. Connect the flask to a reflux condenser, heat on a water bath for 1 h, and allow to cool to room temperature. Disconnect the flask from the condenser, transfer the contents of the conical flask to a 100-mL volumetric flask with the aid of water, and dilute with water to volume. Pass through a dry filter, discarding the first 10 mL of the filtrate. To 50.0 mL of the subsequent filtrate add 5 mL of nitric acid and 2 mL of ferric ammonium sulfate TS, and titrate the excess silver nitrate with 0.1 N ammonium thiocyanate VS. Perform a blank determination, using 5 mL of alcohol in place of the Sample solution, and make any necessary correction. Each mL of 0.1 N silver nitrate is equivalent to 4.958 mg of C4H5NS. Acceptance criteria: 93.0%105.0% SPECIFIC TESTS SPECIFIC GRAVITY 841: 1.0131.020 REFRACTIVE INDEX 831: 1.5271.531 at 20 DISTILLING RANGE, Method I 721: 148154 LIMIT OF PHENOLS Sample solution: Dilute a 1-mL sample with 4 mL of alcohol. Analysis: Add 1 drop of ferric chloride TS to the Sample solution. Acceptance criteria: A blue color is not produced immediately. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Suitability requirements Resolution: NLT 5 between the analyte and internal standard Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity of C5H4N4O in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of allopurinol to hypoxanthine from the Sample solution = peak response ratio of allopurinol to RS hypoxanthine from the Standard solution = concentration of USP Allopurinol RS in the CS Standard solution (mg/mL) = nominal concentration of allopurinol in the CU Sample solution (mg/mL) Acceptance criteria: 93.0%107.0% RU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 2: 75 rpm Time: 45 min Detector: UV 250 nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Standard stock solution: Transfer 40 mg of USP Allopurinol RS to a 200-mL volumetric flask. Add 10 mL of 0.1 N sodium hydroxide, sonicate for about 2 min, and shake by mechanical means for about 10 min. Dilute with Medium to volume. Standard solution: Dilute the Standard stock solution with Medium to obtain a solution having a concentration similar to that expected in the solution under test. Analysis: Determine the amount of C5H4N4O dissolved by using UV absorption at the wavelength of maximum absorbance at about 250 nm on filtered portions of the solution under test, suitably diluted with Medium, in comparison to the Standard solution. Tolerances: NLT 75% (Q) of the labeled amount of C5H4N4O. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Allopurinol RS
Aloe
(Comment on this Monograph)id=m1540=Aloe=A-Monos.pdf) DEFINITION Aloe is the dried latex of the leaves of Aloe barbadensis Miller (Aloe vera Linn ), known in commerce as Cura ao Aloe, or of e c Aloe ferox Miller and hybrids of this species with Aloe africana Miller and Aloe spicata Baker, known in commerce as Cape Aloe (Fam. Liliaceae). Aloe yields NLT 50.0% of water-soluble extractive. IDENTIFICATION A. Powdered Aloe dissolves in nitric acid with effervescence, forming a reddish-brown to brown-or-green solution.
Allyl Isothiocyanate
(Comment on this Monograph)id=m1480=Allyl Isothiocyanate=A-Monos.pdf)
C4H5NS 3-Isothiocyanato-1-propene; Isothiocyanic acid allyl ester [57-06-7].
99.16
USP 32
B. PROCEDURE Sample: 1 g, finely powdered Analysis: Mix the Sample with 25 mL of cold water. Shake the mixture occasionally during 2 h, filter, and wash the filter and residue with sufficient cold water to make the filtrate measure 100 mL. Acceptance criteria: The color of the filtrate, viewed in the bulb of a 100-mL volumetric flask, is dark orange with Cura ao Aloe, and greenish yellow with Cape Aloe. The c filtrate darkens on standing. [NOTEReserve the filtrate for Procedures C and D.] C. PROCEDURE Sample: 5 mL of the filtrate obtained in Identification, Procedure B Analysis: Add 2 mL of nitric acid to the Sample. Acceptance criteria: The mixture exhibits a reddish-orange color with Cura ao Aloe, and a reddish-brown color which c changes rapidly to green with Cape Aloe. D. PROCEDURE Sample: 10 mL of the filtrate obtained in Identification, Procedure B Analysis: Mix the Sample with 2 mL of ammonium hydroxide. Acceptance criteria: The mixture exhibits an amber color with Cape Aloe, and a dark amber color with Cura ao Aloe. c ASSAY PROCEDURE Sample: 2 g Analysis: Macerate the Sample in 70 mL of water in a suitable flask. Shake the mixture during 8 h at 30-min intervals, and allow it to stand for 16 h without shaking. Filter, and wash the flask and residue with small portions of water, passing the washings through the filter, until the filtrate measures 100.0 mL. Evaporate a 50-mL aliquot of the filtrate in a tared dish on a steam bath to dryness, and dry at 110 to constant weight. Acceptance criteria: NLT 50.0% of the weight of Aloe taken SPECIFIC TESTS WATER DETERMINATION, Method III 921 Sample: Use a powdered sample. (If the Aloe is not powdered, crush it in a mortar until it passes through a no. 40 sieve, and mix the ground material before weighing the sample.) Analysis: Dry at 105 for 5 h Acceptance criteria: NMT 12.0% ARTICLES OF BOTANICAL ORIGIN, Total Ash 561 Acceptance criteria: NMT 4.0% ALCOHOL-INSOLUBLE SUBSTANCES Sample: 1 g of powdered Aloe Analysis: Add the Sample to 50 mL of alcohol in a flask. Heat the mixture to boiling, and maintain at incipient boiling for 15 min, replacing any loss by evaporation. Remove from the heat, and shake the mixture at intervals during 1 h. Pass through a small dried and tared filter paper or a dried and tared filtering crucible, and wash the residue on the filter with alcohol until the last washing is colorless. Dry the residue at 105 to constant weight. Acceptance criteria: NMT 10.0% of the weight of Aloe taken BOTANIC CHARACTERISTICS Cura ao aloe: Brownish black, opaque masses. Its fractured c surface is uneven, waxy, and somewhat resinous. Cape aloe: Dusky to dark brown irregular masses, the surfaces of which are often covered with a yellowish powder. Its fracture is smooth and glassy. Powdered aloe: Yellow, yellowish brown to olive-brown in color. When mounted in a bland expressed oil, it appears as greenish yellow to reddish brown angular or irregular
Official Monographs / Alprazolam 87

fragments, the hues of which depend to some extent upon the thickness of the fragments.
Alprazolam
(Comment on this Monograph)id=m1640=Alprazolam=AMonos.pdf)
308.76 C17H13ClN4 4H-[1,2,4]Triazolo[4,3-][1,4]benzodiazepine, 8-chloro-1methyl-6-phenyl-; 8-Chloro-1-methyl-6-phenyl-4H-s-triazolo[4,3-] [1,4]benzodiazepine [28981-97-7]. DEFINITION Alprazolam contains NLT 98.0% and NMT 102.0% of C17H13ClN4. [CAUTIONCare should be taken to prevent inhaling particles of Alprazolam and exposing the skin to it.] IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Sample solution: 4 g/mL in alcohol Analytical wavelength: 220 nm Acceptance criteria: Absorptivities calculated on the dried basis, do not differ by more than 3.0%. ASSAY PROCEDURE Diluent: Acetonitrile and water (1:1) Buffer: 1.36 mg/mL of monobasic potassium phosphate (KH2 PO4) in water Mobile phase: Acetonitrile and Buffer (1:1) Standard solution: 25 g/mL of USP Alprazolam RS in Diluent [NOTEThe solution is stable for 48 h at room temperature when stored in closed containers.] Sample solution: 25 g/mL of Alprazolam in Diluent. [NOTESonication for 1 min may be used to aid dissolution. The solution is stable for 48 h at room temperature when stored in closed containers.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 231 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the quantity, in percentage, of C17H13ClN4 in the portion of Alprazolam taken: Result = (rU/rS) (CS/CU) 100 rU = peak area from the Sample solution
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rS CS
Impurity Table 1 (continued) Relative Retention Time 4.0 Relative Response Factor 1.0 1.0
USP 32
= peak area from the Standard solution = concentration of USP Alprazolam RS in the Standard solution (mg/mL) CU = concentration of alprazolam in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Acceptance criteria: NMT 0.5% HEAVY METALS, Method II 231 Acceptance criteria: NMT 20 ppm Organic Impurities PROCEDURE Diluent: Prepare as directed in the Assay. Buffer: Prepare as directed in the Assay. Mobile phase: Prepare as directed in the Assay. System suitability solution: 20 g/mL each of USP Alprazolam RS, USP Alprazolam Related Compound A RS, and USP 2-Amino-5-chlorobenzophenone RS in Diluent Sample solution: 0.25 mg/mL of alprazolam in Diluent [NOTESonication for 1 min may be used to aid dissolution. When stored in closed containers, the Sample solution is stable for 24 h at room temperature.] Standard solution: 0.25 g/mL of USP Alprazolam RS in Diluent [NOTEWhen stored in closed containers, the Standard solution is stable for 48 h at room temperature.] Chromatographic system (See Chromatography 621.) Proceed as directed in the Assay. System suitability Samples: Standard solution and System suitability solution [NOTEFor relative retention times, see Impurity Table 1.] Suitability requirements Resolution: NMT 2.0 between alprazolam Related compound A and alprazolam, System sutiability solution Relative standard deviation: NMT 5.0, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in each of the related compounds and any of the unspecified impurities: Result = (rU/rS) (CS/CU) (1/F) 100 = peak response for each impurity in the Sample solution = peak response for alprazolam from the Standard rS solution = concentration of USP Alprazolam RS in the CS Standard solution (mg/mL) = concentration of alprazolam in the Sample CU solution F = relative response factor for the corresponding impurity from Impurity Table 1 Acceptance criteria Individual impurities: See Impurity Table 1 Total impurities: NMT 1.0% rU
Impurity Table 1 Relative Retention Time 0.8 1.0 Relative Response Factor 0.76 1.0 Acceptance Criteria, NMT (%) 0.15
Name 2-Amino-5chlorobenzophenone Individual unspecified impurity
Acceptance Criteria, NMT (%) 0.15 0.10
SPECIFIC TESTS LOSS ON DRYING 731: Dry at a pressure of NMT 5 mm of mercury at 60 for 16 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Alprazolam RS
Alprazolam Oral Suspension

(Comment on this Monograph)id=m1377=Alprazolam Oral Suspension=A-Monos.pdf) DEFINITION Alprazolam Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of alprazolam (C17H13ClN4). Prepare Alprazolam Oral Suspension 1 mg/mL as follows (see Pharmaceutical CompoundingNonsterile Solutions 795).
Alprazolam Vehicle: a mixture of Vehicle for Oral Solution (regular or sugar-free), NF, and Vehicle for Oral Suspension, NF (1:1) To make 100 mg A sufficient quantity
100 mL
Comminute Tablets in a suitable mortar to a fine powder, or add Alprazolam powder. Add about 20 mL of the Vehicle, and mix until a uniform paste is formed. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add sufficient Vehicle to bring to final volume, and mix well. ASSAY PROCEDURE Solution A: 0.04 M sodium acetate solution. Adjust with glacial acetic acid to a pH of 2.4. Mobile phase: Methanol, acetonitrile, and Solution A (45:8:47) Standard solution: 20 g/mL of USP Alprazolam RS in Mobile phase Sample solution: Agitate the container of Alprazolam Oral Suspension for 30 min on a rotating mixer, remove a 5-mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the sample from the freezer, allow it to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 1.0 mL of the sample into a 50-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.)
Name Alprazolam related compound A Alprazolam
USP 32
Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 0.6 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 1.4% Retention time: 10 min Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H13ClN4 in each mL of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Alprazolam RS in the Standard solution (g/mL) = nominal concentration of the Sample solution CU (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.05.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at controlled room temperature, or under refrigeration. LABELING: Label it to state that it is to be well-shaken before use, and to state the beyond-use date. Beyond-Use Date: 60 days after the day on which it was compounded USP REFERENCE STANDARDS 11 USP Alprazolam RS rU rS CS
Official Monographs / Alprazolam 89

891, 826, 779, 746, 696, and 658 wavenumbers in the region of 975600 cm1. ASSAY PROCEDURE Mobile phase: Acetonitrile, chloroform, butyl alcohol, glacial acetic acid, and water (850:80:50:0.5:20) Internal standard solution: 0.25 mg/mL of triazolam in acetonitrile Standard stock solution: 0.25 mg/mL of USP Alprazolam RS in Internal standard solution Standard solution: 25 g/mL of USP Alprazolam RS from Standard stock solution in acetonitrile Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a quantity of the powder, equivalent to about 5 mg of alprazolam, to a 200-mL volumetric flask. Transfer 2 mL of water and 20 mL of Internal standard solution, shake vigorously for 10 min, and dilute with acetonitrile to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 30-cm; packing L3 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between the internal standard and alprazolam Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the quantity, as a percentage, of C17H13ClN4 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100
Alprazolam Tablets
(Comment on this Monograph)id=m1647=Alprazolam Tablets=A-Monos.pdf) DEFINITION Alprazolam Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of alprazolam (C17H13ClN4). IDENTIFICATION A. INFRARED ABSORPTION Sample: Finely powdered Tablets, equivalent to 15 mg of alprazolam Analysis: Dissolve the Sample in 10 mL of 10 mg/mL of sodium carbonate solution. Add 15 mL of chloroform, and shake vigorously for 30 min. Centrifuge, withdraw the aqueous layer, and transfer the chloroform to a clean container. Add 200 mg of potassium bromide. Evaporate the chloroform from this mixture to dryness, and dry the dispersion in vacuum at 60 for 24 h. Grind this dispersion into a fine powder. Prepare a suitable pellet for testing by placing 100 mg of dried potassium bromide into a die. Sprinkle 20 mg of the finely ground alprazolampotassium bromide dispersion onto the dried potassium bromide layer, and cover with another specimen of 100 mg of dried potassium bromide. Acceptance criteria: The IR absorption spectrum of the potassium bromide dispersion so obtained exhibits maxima characteristic of alprazolam, as compared to that of a similar preparation of USP Alprazolam RS, at the following wavenumbers: at 1609, 1578, 1566, 1539, 1487, and 1379 wavenumbers in the region of 16501300 cm1; at 932,
= peak area response ratio of the alprazolam peak relative to the internal standard peak from the Sample solution = peak area response ratio of the alprazolam peak RS relative to the internal standard peak from the Standard solution = concentration of USP Alprazolam RS in the CS Standard solution (mg/mL) = nominal concentration of the Sample solution CU (Tablets/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Solution A: 80 g/L of monobasic potassium phosphate and 20 g/L dibasic potassium phosphate in water. Add, with mixing, phosphoric acid or potassium hydroxide solution (45 in 100), as necessary to adjust the solution such that, when this is diluted 1 in 10 with water, the resulting solution has a pH of 6.0 0.1. Buffer: 1-in-10 dilution of Solution A in water to obtain a solution having a pH of 6.0 0.1 Medium: Buffer; 500 mL Apparatus 1: 100 rpm Time: 30 min [NOTEDetermine the amount of C17H13ClN4 dissolved using the following method.] Mobile phase: Acetonitrile, tetrahydrofuran, and Buffer (35:5:60) Standard stock solution: 0.05 mg/mL of USP Alprazolam RS in methanol
RU
90
Alprazolam / Official Monographs

V
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= volume of Internal standard solution used to prepare the Sample solution L = label claim (mg/Tablet) Acceptance criteria: Meet the requirements of the test for Content Uniformity ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Alprazolam RS
Standard solution: Add 50 mL of Solution A and 250 mL of water to a 500-mL flask. Add to the flask 5.0 mL of Standard stock solution for every 0.25 mg of alprazolam contained in the Tablet being assayed. Dilute with water to volume. Sample solution: Sample per 711 Dissolution. Dilute with Medium to a concentration that is similar to the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 10-cm; packing L7 Flow rate: 1 mL/min System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 500 theoretical plates Relative standard deviation: NMT 3.0% for replicate injections Analysis Samples: Filtered portions of the solution from the Dissolution vessel and Standard solution Calculate the quantity of C17H13ClN4 dissolved based on the peak responses from the solution under test and the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C17 H13 ClN4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905 Mobile phase: Acetonitrile, chloroform, butyl alcohol, glacial acetic acid, and water (850:80:50:0.5:20) Internal standard solution: 32 g/mL of triazolam in acetonitrile Standard solution: 25 g/mL of USP Alprazolam RS in Internal standard solution Sample solution: Transfer 1 Tablet to a container. Add 0.4 mL of water directly onto the Tablet, allow the Tablet to stand for 2 min, and then swirl the container to disperse the Tablet. For every 0.25 mg of alprazolam contained in the Tablet, add 10.0 mL of Internal standard solution to the container. Shake, and centrifuge if necessary. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 30-cm; packing L3 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between the internal standard and alprazolam Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H13ClN4 in the Tablet taken: Result = (RU/RS) C V 100/L RU RS C = peak area response ratio of the alprazolam peak relative to the internal standard peak from the Sample solution = peak area response ratio of the alprazolam peak relative to the internal standard peak from the Standard solution = concentration of USP Alprazolam RS in the Standard solution
Alprostadil
(Comment on this Monograph)id=m1658=Alprostadil=AMonos.pdf)
354.48 C20H34O5 Prost-13-en-1-oic acid, 11,15-dihydroxy-9-oxo-, (11,13E, 15S)-; (1R, 2R, 3R)-3-Hydroxy-2-[(E)-(3S)-3-hydroxy-1-octenyl]-5oxocyclopentane heptanoic acid [745-65-3]. DEFINITION Alprostadil contains NLT 95.0% and NMT 105.0% of C20H34O5, calculated on the anhydrous basis. [CAUTIONGreat care should be taken to prevent inhaling particles of Alprostadil and exposing the skin to it.] IDENTIFICATION INFRARED ABSORPTION 197M ASSAY PROCEDURE [NOTEUse freshly prepared solutions.] Mobile phase: Methanol, acetonitrile, and 0.1 M monobasic potassium phosphate (2:1:2). Adjust with phosphoric acid to a pH of 3.0. Internal standard solution: 0.05 mg/mL of ethylparaben in methanol and water (9:1) Standard stock solution: 0.3 mg/mL of USP Alprostadil RS in methanol and water (9:1) Standard solution: Internal standard solution and Standard stock solution (1:2) System suitability stock solution: 4.5 g/mL of prostaglandin A1 from USP Prostaglandin A1 RS in Standard solution System suitability solution: Internal standard solution and System suitability stock solution (1:2) Sample stock solution: 0.3 mg/mL of Alprostadil in a mixture of methanol and water (9:1) Sample solution: Combine 1.0 mL of Internal standard solution and 2.0 mL of Sample stock solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Photodiode array or equivalent capable of detecting UV wavelengths of 200300 nm
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Analytical wavelength: 200 nm Column: 4.6-mm 25-cm; packing L1 Column temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 7.5 between prostaglandin A1 and alprostadil, and NLT 2.0 between prostaglandin A1 and ethylparaben Relative standard deviation: NMT 2.0%, determined from the peak area ratio of alprostadil to ethylparaben for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C20H34O5 in the portion taken: Result = (RU/RS) (CS/CU) 100 = peak area ratio of Alprostadil to ethylparaben in the Sample solution = peak area ratio of alprostadil to ethylparaben in RS the Standard solution = concentration of USP Alprostadil RS in the CS Standard solution (mg/mL) = concentration of Alprostadil in the Sample CU solution (mg/mL) Acceptance criteria: 95.0%105.0% RU IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Sample: 0.3 g Acceptance criteria: NMT 0.5% LIMIT OF CHROMIUM Standard stock solution: 3.04 g/mL of chromium trichloride in 0.05 M nitric acid Standard solution: 20 ng/mL of chromium (Cr) in alcohol by diluting 2 mL of Standard stock solution to 100 mL with alcohol Sample solution: 1 mg/mL of Alprostadil in alcohol Blank: Alcohol Spectrometric conditions (See Spectrophotometetry and Light-Scattering 851.) Mode: Graphite furnace atomic absorption spectrophotometer (equipped with a chromium hollowcathode lamp) Analytical wavelength: 357.9 nm Drying temperature: 100 Ashing temperature: 1000 Atomization temperature: 2700 Injection size: 20 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of Cr in the portion of C20H34O5 taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of chromium in the Standard solution (mg/mL) = concentration of Alprostadil in the Sample CU solution (mg/mL) Acceptance criteria: NMT 20 ppm LIMIT OF RHODIUM Standard stock solution: 100 g/mL of rhodium in 1.2 M hydrochloric acid by diluting rhodium chloride hydrate Standard solution: 50 ng/mL of rhodium (Rh) in alcohol from Standard stock solution AU AS CS
Official Monographs / Alprostadil 91

Sample solution: 2 mg/mL of Alprostadil in alcohol Blank: Alcohol Spectrometric conditions (See Spectrophotometetry and Light-Scattering 851.) Mode: Graphite furnace atomic absorption spectrophotometer (equipped with a rhodium hollowcathode lamp) Analytical wavelength: 343.5 nm Drying temperature: 100 Ashing temperature: 1000 Atomization temperature: 2800 Injection size: 20 L Analysis Samples: Standard solution and Sample solution Calculate the percentage of Rh in the portion of Alprostadil taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of rhodium in the Standard solution (mg/mL) = concentration of Alprostadil in the Sample CU solution (mg/mL) Acceptance criteria: NMT 20 ppm Organic Impurities PROCEDURE 1: LIMIT OF FOREIGN PROSTAGLANDINS [NOTEUse freshly prepared solutions. ] Mobile phase: Methanol, acetonitrile, and 0.1 M monobasic potassium phosphate (2:1:2). Adjust with phosphoric acid to a pH of 3.0. Standard solution: 6 g/mL of USP Alprostadil RS,15 g/mL of USP Prostaglandin A1 RS, and 6 g/mL of USP Prostaglandin B1 RS in methanol and water (9:1) Sample solution: 3.0 mg/mL of Alprostadil in methanol and water (9:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Photodiode array detector or equivalent capable of detecting UV wavelengths of 200300 nm Analytical wavelengths: Prostaglandin A1 at 224 nm, prostaglandin B1 at 280 nm, and all other foreign prostaglandin impurities at 200 nm. Column: 4.6-mm 25-cm; packing L1 Column temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 7.5 between prostaglandin A1 and alprostadil Relative standard deviation: NMT 4.0%, determined from the peaks at their respective wavelength for replicate injections Analysis 1 Samples: Standard solution and Sample solution Calculate the percentage of prostaglandin A1 and prostaglandin B1 in the portion of C20H34O5 taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response of prostaglandin A1 or prostaglandin A2 from the Sample solution = peak response of prostaglandin A1 or prostaglandin A2 from the Standard solution = concentration of USP Prostaglandin A1 RS or USP Prostaglandin B1 RS in the Standard solution (mg/mL) AU AS CS
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Alprostadil / Official Monographs

= concentration of Alprostadil in the Sample solution (mg/mL) Acceptance criteria 1 Prostaglandin A1: NMT 1.5% Prostaglandin B1: NMT 0.1% Analysis 2: Calculate the percentage of each impurity occurring at 200 nm and eluting before prostaglandin A1 in the portion of C20H34O5 taken: Result = (rU/rS) (CS/CU) 100 = peak response for each impurity of the Sample solution = peak response for alprostadil of the Standard rS solution = concentration of USP Alprostadil RS in the CS Standard solution (mg/mL) = concentration of Alprostadil in the Sample CU solution (mg/mL) Acceptance criteria 2: NMT 0.9% of any foreign prostaglandin impurity eluting before prostaglandin A1 Analysis 3: Calculate the percentage of any impurity having a relative retention time of 0.6, relative to the prostaglandin A1 peak detected at 224 nm, in the portion of C20H34O5 taken: Result = (rU/rS) (CS/CU) 100 rU rU CU
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Relative standard deviation: NMT 2.0%, determined from the main peak in the Standard solution (multiple injections) Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity occurring at 200 nm and eluting after prostaglandin A1, excluding prostaglandin B1, in the portion of C20H34O5 taken: Result = (rU/rS) (CS/CU) 100 = peak response for each impurity of the Sample solution rS = peak response for alprostadil of the Standard solution CS = concentration of USP Alprostadil RS in the Standard solution (mg/mL) CU = concentration of Alprostadil in the Sample solution (mg/mL) Acceptance criteria: The sum of the peaks having relative retention times of 2.0 and 2.3 is NMT 0.6%; any other foreign prostaglandin impurity eluting after prostaglanin A1 is NMT 0.9%. Total impurities for Procedure 1 and Procedure 2: NMT 2.0 % SPECIFIC TESTS WATER DETERMINATION, Method I 921 Sample: 0.5 g Acceptance criteria: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a refrigerator. USP REFERENCE STANDARDS 11 USP Alprostadil RS USP Prostaglandin A1 RS USP Prostaglandin B1 RS
= peak response for any impurity having a relative retention time of 0.6, relative to the prostaglandin A1 peak, from the Sample solution rS = peak response for prostaglandin A1 from the Standard solution CS = concentration of USP Prostaglandin A1 RS in the Standard solution (mg/mL) CU = concentration of Alprostadil in the Sample solution (mg/mL) Acceptance criteria 3: NMT 0.9% of any impurity having a relative retention time of 0.6, relative to the prostaglandin A1 peak PROCEDURE 2: LIMIT OF FOREIGN PROSTAGLANDINS Mobile phase: Methanol, acetonitrile, and 0.02 M monobasic potassium phosphate (2:1:1). Adjust with phosphoric acid to a pH of 3. Standard solution: 10 g/mL of USP Alprostadil RS in acetonitrile and water (1:1) Sample solution: 5.0 mg/mL of Alprostadil in acetonitrile and water (1:1) [NOTESonicate if necessary.] System suitability solution: 6 g/mL of USP Alprostadil RS, 15 g/mL of USP Prostaglandin A1 RS, and 6 g/mL of USP Prostaglandin B1 RS in methanol and water (9:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Photodiode array detector or equivalent, capable of detecting UV wavelengths of 200300 nm Analytical wavelengths: 224 nm for prostaglandin A1, 280 nm for prostaglandin B1 , and 200 nm for all other prostaglandins Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for prostaglandin A1 and alprostadil in the chromatogram of the System suitability solution are 1.2 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between prostaglandin A1 and alprostadil for the Identification solution rU
Alprostadil Injection
(Comment on this Monograph)id=m1660=Alprostadil Injection=A-Monos.pdf) DEFINITION Alprostadil Injection is a sterile solution of Alprostadil in Dehydrated Alcohol. It contains NLT 90.0% and NMT 115.0% of the labeled amount of alprostadil (C20H34O5). IDENTIFICATION INFRARED ABSORPTION 197K Standard: A preparation similar to that of the Sample, using USP Alprostadil RS in dehydrated alcohol Sample: Dry an amount of Injection, equivalent to 2 mg of alprostadil, on 500 mg of spectroscopic grade potassium bromide at about 4050 under vacuum. Prepare a pellet from this mixture. ASSAY PROCEDURE Mobile phase: Methylene chloride, 1,3-butanediol, and water (1000:6:0.5) Internal standard solution: 50 g/mL of ethylparaben in methylene chloride Standard stock solution: 0.5 mg/mL of USP Alprostadil RS in dehydrated alcohol Standard solution: Gently evaporate a 0.5-mL portion of the Standard stock solution to dryness with a stream of nitrogen. Add 150 mL of a freshly prepared 5 mg/mL of diisopropylethylamine in acetonitrile to the container, rinse the inside of the container with this solution, and swirl. Cap
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and sonicate to dissolve. Heat the container at 45 for 45 min, swirling occasionally. Sonicate again after heating is complete. [NOTEIf the entire sample does not dissolve, the specimen should be discarded.] Evaporate the solution using a stream of nitrogen, add 2.0 mL of Internal standard solution, and sonicate to dissolve. [NOTEIf incomplete dissolution is still observed, discard the specimen.] Sample solution: Pool the contents of several containers of the Injection, and gently evaporate measured volume, equivalent to 0.25 mg of alprostadil, to dryness using a stream of nitrogen. Add 150 L of a freshly prepared solution of 40 mg/mL -bromo-2-acetonaphthone in acetonitrile, rinse the inside of the container with this solution, and swirl. Gently evaporate a 0.5-mL portion of the Standard stock solution to dryness with a stream of nitrogen. Add 150 mL of a freshly prepared 5 g/mL diisopropylethylamine in acetonitrile to the container, rinse the inside of the container with this solution, and swirl. Cap and sonicate to dissolve. Heat the container at 45 for 45 min, swirling occasionally. Sonicate again after heating is complete. [NOTEIf the entire sample does not dissolve, the specimen should be discarded.] Evaporate the solution using a stream of nitrogen, add 2.0 mL of Internal standard solution, and sonicate to dissolve. [NOTEIf incomplete dissolution is still observed, discard the specimen.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.4-mm 25-cm; packing L18 Flow rate: 1.5 mL/min System suitability Sample: Standard solution [NOTEThe relative retention times for ethylparaben and alprostadil are about 0.4 and 1.0, respectively.] Suitability requirements Resolution: NLT 9.0 between alprostadil and the internal standard Relative standard deviation: NMT 2.5% Analysis Samples: Standard solution and Sample Solution Calculate the quantity, as a percentage, of C20H34O5 in the volume of Injection taken: Result = (RU/RS) (CS/CU) 100/L = peak response ratio of alprostadil to the internal standard from the Sample solution RS = peak response ratio of alprostadil to the internal standard from the Standard solution CS = concentration of USP Alprostadil RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (unit/mL) L = label claim (mg/unit) Acceptance criteria: 90.0%115.0% RU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5 USP Endotoxin Units/100 g of alprostadil. STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined for Membrane Filtration. WATER DETERMINATION, Method I 921: NMT 0.4% OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose containers, preferably of Type I glass. Store in a refrigerator. USP REFERENCE STANDARDS 11 USP Alprostadil RS USP Endotoxin RS
Official Monographs / Alteplase 93
Alteplase
(Comment on this Monograph)id=m1670=Alteplase=AMonos.pdf)
C2569H3894N746O781S40 [105857-23-6].
59,007.61
DEFINITION Alteplase is a highly purified glycosylated serine protease with fibrin-binding properties and plasminogen-specific proteolytic activities. It is produced by recombinant DNA synthesis in mammalian cell culture. It has a biological potency of NLT 90.0% and NMT 115.0% of the potency stated on the label, the potency being 580,000 USP Alteplase Units/mg of protein. The presence of host cell DNA and host cell protein impurities in Alteplase is process specific; the limits of these impurities are determined by validated methods. IDENTIFICATION A. PROCEDURE Sample solution: 1.02.5 mg/mL Alteplase Standard solution: Prepare similarly to the Sample solution. Analysis Samples: Standard solution and Sample solution To each of three test tubes, transfer 1 mL of 0.5 mg/mL HD-isoleucyl-prolyl-arginyl-p-nitroaniline dihydrochloride. Separately transfer 200 L of the Sample solution and 200 L of the Standard solution to two of the test tubes, and to the third test tube, add 200 L of 0.2 M arginine solution that has been adjusted with phosphoric acid (negative control) to a pH of 7.3. Mix the solutions in the test tubes, and allow to stand for 1 min. Acceptance criteria: A yellow color is produced in the solutions from the Sample solution and the Standard solution, while no yellow color is produced in the negative control. ASSAY PROCEDURE FOR BIOLOGICAL POTENCY Buffer: 1.38 mg/mL of monobasic sodium phosphate, 7.10 mg/mL of anhydrous dibasic sodium phosphate, 0.20 mg/mL of sodium azide, and 0.10 mg/mL of polysorbate 80 in water Human thrombin solution: 33 U.S. Units/mL in Buffer [NOTEUnits are in terms of the U.S. Standard Thrombin.] Human fibrinogen solution: 2 mg/mL of human fibrinogen in Buffer Human plasminogen solution: 1 mg/mL of human plasminogen in Buffer Standard stock solution: 1.0 mg/mL (580,000 USP Alteplase Units) of USP Alteplase RS Standard solution: Dilute volumes of Standard stock solution to obtain a series of five Standard solutions having known concentrations ranging from 145 to 9.3 USP Alteplase Units/mL. Sample stock solution: 1.0 mg/mL of Alteplase Sample solution: Dilute a volume of Sample stock solution with Buffer to obtain a series of dilutions of 1:20,000, 1:10,000, and 1:5,000. Analysis: To a set of labeled glass test tubes add 0.5 mL of Human thrombin solution. To separate test tubes add 0.5 mL of each of the Standard solutions and Sample solutions, and store on ice. To a second set of labeled glass tubes, add 20 L of Human plasminogen solution and 1 mL of Human fibrinogen solution, and store on ice. Beginning with the thrombin mixture tubes containing the Standard solution
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with the lowest number of USP Units/mL, record the time, and separately add 200 L of each of the thrombin mixtures to the test tubes containing the plasminogen-fibrinogen mixture. Using a vortex mixer, intermittently mix the contents of each tube for a total of 15 s, and carefully place into a rack in a 37 circulating water bath. A visually turbid clot forms within 30 s, followed by the formation of bubbles within the clot. Record the clot lysis time (tcl) from the first addition of the Alteplase solution to the last bubble to rise to the surface. Using a least squares fit, determine the equation of the line using the log values of the standard concentration, in USP Alteplase Units/mL, versus the log values of their clot lysis times in seconds taken: log t = m(log US) + b = time to bubble release (s) = slope of the line = activity of the Standard solution (USP Alteplase Units/mL) b = y-intercept of the line [NOTEThe correlation coefficient is NLT 0.9900.] From the line equation and using the log of the clot lysis time for the Sample solution, calculate the log of the activity (UA): log UA = [(log t)-b]/m Calculate the alteplase activity in USP Alteplase Units/mL taken: Result = D(10logU) D = dilution factor for the Sample solution Calculate the specific activity in the portion of Alteplase taken: Result = UA/P t m US
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Running buffer: 3.03 mg/mL of tris(hydroxymethyl)aminomethane and 14.26 mg/mL of glycine in sodium dodecyl sulfate (1 in 1000) Carboxymethylation buffer: 480 mg/mL of urea, 44 mg/mL of tris(hydroxymethyl)aminomethane, and 1.2 mg/mL of edetic acid in water. Adjust with hydrochloric acid, if necessary, to a pH of 8.6. Gel: Prepare a 10% T (total acrylamide)0.25% C (crosslinked bisacrylamide) resolving gel containing 0.1% sodium dodecyl sulfate, 0.375 M tris(hydroxymethyl)aminomethane hydrochloride, and 0.05 M tris(hydroxymethyl)aminomethane. Arginine solution: 34.8 mg/mL of arginine in water. Adjust with phosphoric acid to a pH of 7.3. Standard stock solution: 1 mg/mL of USP Alteplase RS in water Standard solution: 0.25 mg/mL USP Alteplase RS from Standard stock solution in Arginine solution. Heat 0.5 mL of this solution with 116 L of SDS buffer and 10 L of 1 M dithiothreitol at 80 for 2 min. Carboxymethylated standard solution: Dilute 1.0 mL of Standard stock solution with 1 mL of Carboxymethylation buffer, and adjust with 1 M sodium hydroxide to a pH of 8.5. Add 20 L of 1 M dithiothreitol, and incubate at 37 for 60 min. Add 100 L of 1 M iodoacetic acid, and incubate in the dark for 20 min. Desalt by passing the solution through a chromatographic column containing fine gel chromatographic packing equilibrated with a buffer solution containing, in each mL 20 mg of sodium dodecyl sulfate, 100 mg of glycerol, 1.42 mg of tris(hydroxymethyl)aminomethane hydrochloride, and 0.85 mg of tris(hydroxymethyl)aminomethane. Collect the protein fraction of the preparation by elution with the same buffer, and add 20 L of 1 M dithiothreitol. Adjust the protein concentration to about 0.2 mg/mL with a buffer solution containing, in each mL 20 mg of sodium dodecyl sulfate, 100 mg of glycerol, 1.42 mg of tris(hydroxymethyl)aminomethane hydrochloride, 0.85 mg of tris(hydroxymethyl)aminomethane, 1.06 mg of dithiothreitol, 0.05 mg of bromophenol blue, and 0.05 mg of xylene cyanole FF. Sample stock solution, Sample solution, and Carboxymethylated sample solution: Using Alteplase, proceed as directed for Standard stock solution, Standard solution, and Carboxymethylated standard solution. Molecular weight standard solution: Use a commercially available preparation of low molecular weight protein standards (10,000 to 100,000 Da) at 2 mg/mL. Mix 990 L of Diluted SDS buffer and 10 L of the molecular weight standard mixture. Control solution: Prepare a control solution of bovine serum albumin containing 10 g/ mL. For a 10 ng/25 L load, mix 600 L of Diluted SDS buffer and 25 L of the control solution, and heat at 90 for 2 min. For a 2.5 ng per 25 L load, mix 594 L of Diluted SDS buffer and 6 L of the control solution, and heat at 90 for 2 min. Blank: Mix 500 L of water, 126 L of SDS buffer, and 10 L of 1 M dithiothreitol. Analysis Samples: Sample solution, Standard solution, Carboxymethylated sample solution, and Carboxymethylated standard solution, Control solution, Molecular weight standard solution, and Blank. Separately apply equal volumes (about 25 L) of the Sample solution, Standard solution, Carboxymethylated sample solution, and Carboxymethylated standard solution at the 5 g load; apply equal volumes (about 38 L) of the Standard solution and the Carboxymethylated standard solution at the 7.5 g load; and apply the Control
= concentration of protein obtained in the test for Protein Content Acceptance criteria: NLT 90% and NMT 115% of the potency stated on the label, the potency being 580,000 USP Alteplase Units/mg of protein IMPURITIES Organic Impurities PROCEDURE (See Electrophoresis 726.) SDS buffer: 400 mg/mL of glycerol, 5.52 mg/mL of tris(hydroxymethyl)aminomethane hydrochloride, 3.28 mg/mL of tris(hydroxymethyl)aminomethane, 0.20 mg/mL of bromophenol blue, and 0.20 mg/mL of xylene cyanole FF in sodium dodecyl sulfate solution (8 in 100) Diluted SDS buffer: Dilute 1 volume of SDS buffer with four volumes of water. Ammoniacal silver nitrate solution: Transfer 105 mL of sodium hydroxide solution (0.36 in 100) and 7.0 mL of ammonium hydroxide to a 500-mL volumetric flask, and add slowly, with stirring, 20.0 mL of silver nitrate solution (20 in 100). Dilute with water to volume. [NOTEPrepare this solution immediately before use, and protect it from light. This amount of solution is sufficient for two slab gels.] Citric acidformaldehyde solution: To 500 mL of water add 25 mg of citric acid, 0.25 mL of formaldehyde, and 0.025 mL of methanol, omitting the methanol if the formaldehyde is preserved with methanol. [NOTEPrepare this solution fresh at the time of use. This amount of solution is sufficient for two slab gels.]
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solutions at the 10 and 2.5 ng load onto separate lanes of the gel. Apply about 25 L of the Molecular weight standard solution to each side of the gel, and apply about 25 L of the Blank onto a separate lane. Apply the Sample solution and the Standard solution on one half and the Carboxymethylated sample solution and Carboxymethylated standard solution on the other half. Perform the electrophoresis using a constant current of 1.31.5 mAmp/cm of gel length and the Running buffer. Remove the gel from the apparatus 1020 min after the tracking dye starts to move. Place the gel in 250 mL of a solution of 20% alcohol and 6% glacial acetic acid for NLT 1 h, and change the solution every 20 min, leaving the gel to soak overnight following the last change. Perform silver staining of the gel by placing the gel in 250 mL of a solution (1 in 10) in a shallow dish, and shake for about 30 min. Replace the glutaraldehyde solution with distilled water, allow the gel to soak for about 20 min, and then change the water. Repeat for a total of three washings. Transfer the gel to a dish, and cover with 250 mL of Ammoniacal silver nitrate solution. Place the dish on a shaker for about 15 min. Rinse four times with 250 mL of water, rocking the dish for 1 min between rinses. Continue rocking to prevent the gel from sticking and to facilitate washing. Transfer the gel to a clear dish containing 250 mL of Citric acidformaldehyde solution, and rock the dish. Protein bands become visible. When the gel is visibly stained, wash immediately with water, and rinse it repeatedly with water to remove the Citric acidformaldehyde solution. Rinse the gel for NLT 1 h, and dry. Soak cellophane membranes in glycerol solution (2 in 100). Roll a membrane onto a rigid sheet of plastic. Roll the gel onto the membrane, and cover with another membrane. Lay a frame on the edges of the membranes, and clamp it to the rigid plastic sheet. Dismantle the dryer, and cut off excess cellophane when dry (about 24 h). Visually examine the gel under light. Acceptance criteria: The Sample solution exhibits three major bands in the region between 66,000 Da and 31,000 Da, corresponding to the major bands from the Standard solution. The Carboxymethylated sample solution exhibits six major bands in the region between 92,500 Da and 45,000 Da, corresponding to the major bands from the Carboxymethylated standard solution. System suitability: The 2.5 and 10ng Controls must be visible. The nonreduced Control solutions migrate with an apparent molecular weight of slightly less than 66,000 Da, as compared with the molecular weight standards. SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 1 USP Endotoxin Unit/mg of alteplase SINGLE-CHAIN CONTENT Mobile phase: 27.6 g of monobasic sodium phosphate in 1000 mL of sodium dodecyl sulfate solution. Adjust with sodium hydroxide to a pH of 6.8. Dithiothreitol solution: 3.12 mg/mL of dithiothreitol in Mobile phase Standard stock solution: 1 mg/mL of USP Alteplase RS in water Standard solution: Pipet 1 mL of the Standard stock solution into a glass tube, add 3 mL of Dithiothreitol solution, cap the tube, and invert to mix. Heat for 3 to 5 min at about 80. Sample stock solution: 1 mg/mL of Alteplase in water Sample solution: Pipet 1 mL of the Sample stock solution into a glass tube, add 3 mL of Dithiothreitol solution, cap the tube, and invert to mix. Heat for 3 to 5 min at about 80. Chromatographic System (See Chromatography 621, System Suitability.)

Mode: LC Detector: UV 214 nm Column: 7.5-mm 60-cm; packing L25 Flow rate: 0.5 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.1 between the single-chain and twochain alteplase peaks Analysis Samples: Standard solution and Sample solution [NOTEThe major peaks are from single-chain and twochain alteplase and from higher and lower molecular weight species.] Calculate the percentage of single-chain alteplase in the portion of Alteplase taken: Result = (rA/rS) 100 = peak response for single-chain alteplase rA = sum of the responses of all the alteplase peaks rS Acceptance criteria: NLT 60%; no peaks or shoulders in the Sample solution that are not present in the Standard solution are found. PEPTIDE MAPPING Solution A: 6.9 mg/mL of monobasic sodium phosphate in water, adjusted with phosphoric acid to a pH of 2.85 [NOTEMake adjustments if necessary.] Solution B: Acetonitrile Mobile phase: See the gradient table below. [NOTEEquilibrate the system before use with 100% Solution A.]
Time (min) 0 90 120 130 Solution A (%) 100 70 40 40 Solution B (%) 0 30 60 60
Dialysis solution: 480 mg/mL of urea, 44 mg/mL of tris(hydroxymethyl)aminomethane, and 0.88 mg/mL of edetic acid in water [NOTEAdjust with hydrochloric acid to a pH of 8.6.] Standard solution: Prepare a solution containing 1.0 mg/mL of USP Alteplase RS in water. Dialyze 2.0 mL of this solution into the Dialysis solution at room temperature for NLT 12 h. Measure the volume of the solution, and transfer it to a clean test tube. For each mL of solution in the tube, add 10 L of 1 M dithiothreitol. Incubate at room temperature for 4 h, then add 25 L of 1 M iodoacetic acid/mL of the solution, and incubate in the dark for 30 min. Quench the reaction by adding 50 L of 1 M dithiothreitol/mL of the solution. Dialyze the solution against 0.1 M ammonium bicarbonate for 24 h, replacing the 0.1 M ammonium bicarbonate twice during the dialysis period. To 2.0 mL of the dialyzed solution, add 20 g of trypsin, and incubate for 68 h at room temperature. Again add 20 g of trypsin, and incubate for 1618 h for a total of 24 h of incubation of the trypsin-treated solution. [NOTEStore this Standard solution in a freezer.] Sample solution: Using a quantity of Sample, proceed as directed for the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.)
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95% and NMT 111% of the total protein content stated on the label. IDENTIFICATION A. PROCEDURE Arginine solution: 0.2 M Arginine that has been adjusted with phosphoric acid (negative control) to a pH of 7.3 Standard solution: Prepare a concentration similar to that of the Sample solution using USP Alteplase RS. Sample solution: 1.02.5 mg/mL of Alteplase in water Analysis Samples: Sample solution and Standard solution To each of three test tubes transfer 1 mL of 0.5 mg/mL HD-isoleucyl-prolyl-arginyl-p-nitroaniline dihydrochloride containing 0.5 mg/mL. Separately transfer 200 L of the Sample solution and 200 L of the Standard solution to two of the test tubes, and to the third test tube, add 200 L of Arginine solution. Mix the solutions in the three test tubes, and allow to stand for 1 min. Acceptance criteria: A yellow color is produced in the solutions containing the Sample solution and the Standard solution, while no yellow color is produced by the Arginine solution negative control. B. PEPTIDE MAPPING Solution A: 6.9 mg/mL of monobasic sodium phosphate in water, adjusted with phosphoric acid to a pH of 2.85 [NOTEFilter and degas. Make adjustments if necessary.] Solution B: Acetonitrile Mobile phase: See the gradient table below. [NOTEEquilibrate the system before use with 100% Solution A.]
Mode: LC Detector: UV 214 nm Column: 4.6-mm 10-cm; packing L1 Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Requirement 1: NLT 1.5 between peaks 6 and 7 as defined by the USP Alteplase RS Data Sheet Requirement 2: NMT 0.5 min for their baseline widths Analysis Samples: Standard solution, Sample Solution, and a mixture of the Standard solution and the Sample solution (1:1) [NOTEMeasure the responses for NLT 20 major peaks as defined in the USP Alteplase RS Data Sheet.] Acceptance criteria: The retention times of corresponding peaks from the Standard solution and the Sample solution do not differ by more than 0.4 min, and the peak area ratios relative to peak 19 (as shown on the USP Alteplase RS Data Sheet) do not differ by more than 20%. No additional significant peaks or shoulders are found, a significant peak or shoulder being defined as one having a peak area response of NLT 5% of peak 19. PROTEIN CONTENT Arginine solution: 34.8 mg/mL of arginine in water. Adjust with phosphoric acid to a pH of 7.3. Sample stock solution: 1 mg/mL of Alteplase in water Sample solution: Dilute a volume of Sample stock solution with a volume of Arginine solution to obtain a solution having an absorbance value of 0.51.0 at the wavelength of maximum absorbance at about 280 nm. Determine the dilution volume (V). Spectrometric conditions (See Spectroscopy and Light-Scattering 851.) Mode: UV Wavelength range: 240500 nm Analytical wavelengths: 320 nm and peak maxima about 280 nm Cell: 1 cm Blank: Arginine solution Analysis Samples: Sample solution and Blank Calculate the protein content in the portion of Alteplase taken: Result = (Amax A320)/1.9 V Amax A320 V = absorbance value at the wavelength of maximum absorbance (at about 280 nm) = absorbance of the Sample solution at 320 nm = volume of 0.2 M Arginine solution required to prepare the Sample solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in the frozen state at a temperature of 20 or below. USP REFERENCE STANDARDS 11 USP Alteplase RS USP Endotoxin RS
Alteplase for Injection

(Comment on this Monograph)id=m1673=Alteplase for Injection=A-Monos.pdf) DEFINITION Alteplase for Injection is a sterile lyophilized preparation of Alteplase. Its biological activity is NLT 90% and NMT 115% of that stated on the label in USP Alteplase Units. It contains NLT
Dialysis solution: 480 mg/mL of urea, 44 mg/mL of tris(hydroxymethyl)aminomethane, and 0.88 mg/mL of edetic acid in water. [NOTEAdjust with hydrochloric acid to a pH of 8.6.] Standard solution: Prepare a solution containing 1.0 mg/mL of USP Alteplase RS in water. Dialyze 2.0 mL of this solution into the Dialysis solution at room temperature for NLT 12 h. Measure the volume of the solution, and transfer it to a clean test tube. For each mL of solution in the tube, add 10 L of 1 M dithiothreitol. Incubate at room temperature for 4 h, then add 25 L of 1 M iodoacetic acid/mL of the solution, and incubate in the dark for 30 min. Quench the reaction by the addition of 50 L of 1 M dithiothreitol/mL of the solution. Dialyze the solution against 0.1 M ammonium bicarbonate for 24 h, replacing the 0.1 M ammonium bicarbonate twice during the dialysis period. To 2.0 mL of the dialyzed solution, add 20 g of trypsin, and incubate for 68 h at room temperature. Again add 20 g of trypsin, and incubate for 1618 h for a total of 24 h of incubation of the trypsin-treated solution. [NOTEStore in a freezer.] Sample solution: Prepare a solution containing 1.0 mg/mL of alteplase in water. Dialyze 2.0 mL of this solution into the Dialysis solution at room temperature for NLT 12 h. Measure the volume of the solution, and transfer it to a clean test tube. For each mL of solution in the tube, add 10 L of 1 M dithiothreitol. Incubate at room temperature for 4 h, then add 25 L of 1 M iodoacetic acid/mL of the solution, and incubate in the dark for 30 min. Quench the reaction by the
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addition of 50 L of 1 M dithiothreitol/mL of the solution. Dialyze the solution against 0.1 M ammonium bicarbonate for 24 h, replacing the 0.1 M ammonium bicarbonate twice during the dialysis period. To 2.0 mL of the dialyzed solution, add 20 g of trypsin, and incubate for 68 h at room temperature. Again add 20 g of trypsin, and incubate for 1618 h for a total of 24 h of incubation of the trypsin-treated solution. [NOTEStore in a freezer.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 10-cm; packing L1 Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Requirement 1: The resolution between peaks 6 and 7 as defined by the USP Alteplase RS Data Sheet is NLT 1.5 Requirement 2: The width of the peaks at the baseline are NMT 0.5 min. Analysis Samples: Standard solution, Sample Solution,and a mixture of the Standard solution and the Sample solution (1:1) [NOTEMeasure the responses for NLT 20 major peaks as defined in the USP Alteplase RS Data Sheet.] Acceptance criteria: The retention times of corresponding peaks from the Standard solution and the Sample solution do not differ by more than 0.4 min, and the peak area ratios relative to peak 19 (as shown on the USP Alteplase RS Data Sheet) do not differ by more than 20%. No additional significant peaks or shoulders are found, a significant peak or shoulder being defined as one having a peak area response of NLT 5% of peak 19. ASSAY BIOLOGICAL POTENCY [NOTEWhen constituted with water, Alteplase for Injection meets this requirement.] Buffer: 1.38 mg/mL of monobasic sodium phosphate, 7.10 mg/mL of anhydrous dibasic sodium phosphate, 0.20 mg/mL of sodium azide, and 0.10 mg/mL of polysorbate 80 in water Human thrombin solution: 33 U.S. Units/mL in Buffer [NOTEUnits are in terms of the U.S. Standard Thrombin.] Human fibrinogen solution: 2 mg/mL of human fibrinogen in Buffer Human plasminogen solution: 1 mg/mL of human plasminogen in Buffer Standard stock solution: 1.0 mg/mL (580,000 USP Alteplase Units) of USP Alteplase RS Standard solutions: Dilute volumes of Standard stock solution to obtain a series of five Standard solutions having known concentrations ranging from 145 to 9.3 USP Alteplase Units/mL. Sample stock solution: 1.0 mg/mL of Alteplase Sample solutions: Dilute a volume of Buffer to obtain a series of dilutions of 1:20,000, 1:10,000, and 1:5,000. Analysis Samples: Standard solutions and Sample solutions To a set of labeled glass test tubes, add 0.5 mL of Human thrombin solution. To separate test tubes, add 0.5 mL of each of the Standard solutions and Sample solutions, and store on ice. To a second set of labeled glass tubes, add 20 L of Human plasminogen solution and 1 mL of Human fibrinogen solution, and store on ice. Beginning with the thrombin mixture tubes containing the Standard solution with the lowest number of USP Units/mL, record the time, and separately add 200 L of each of the thrombin mixtures to the test tubes containing the

plasminogenfibrinogen mixture. Using a vortex mixer, intermittently mix the contents of each tube for a total of 15 s, and carefully place into a rack in a 37 circulating water bath. A visually turbid clot forms within 30 s, followed by the formation of bubbles within the clot. Record the clot lysis time, tcl, from the first addition of the Alteplase solution to the last bubble to rise to the surface. Using a least squares fit, determine the equation of the line using the log values of the standard concentration, in USP Alteplase Units/mL, versus the log values of their clot lysis times in s taken: log t = m(log US) + b t US = time to bubble release(s) = activity of the Standard solution (USP Alteplase Units/mL) m = slope of the line b = y-intercept of the line [NOTEThe correlation coefficient is NLT 0.9900.] From the line equation and using the log of the clot lysis time for the Sample solution, calculate the log of the activity, UA: log UA = [(log t) b]/m Calculate the alteplase activity, in USP Alteplase Units/mL: Result = D(10log U) D = dilution factor for the Sample solution Calculate the specific activity in the portion of Alteplase: Result = UA/P = concentration of protein obtained in the test Procedure 1: Content of Protein Acceptance criteria: 90.0%115.0% OTHER COMPONENTS PROCEDURE 1: CONTENT OF PROTEIN (See Spectrophotometry and Light-Scattering 851.) Arginine solution: 34.8 mg/mL of arginine [NOTEAdjust with phosphoric acid to a pH of 7.3.] Sample stock solution: 1 mg/mL Sample solution: Dilute a volume of Sample stock solution with a volume of Arginine solution to obtain a solution having an absorbance value of 0.51.0 at the wavelength of maximum absorbance at about 280 nm. Determine the dilution volume, V. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: UV Wavelength range: 240500 nm Analytical wavelengths: 320 nm and peak maxima B at about 280 nm Cell: 1 cm Blank: Arginine solution Analysis Samples: Sample solution and Blank Calculate the protein content in the portion of Alteplase taken: Result = (Amax A320)/F V Amax A320 F V = absorbance value at the wavelength of maximum absorbance = absorbance of the Sample solution at 320 nm = conversion factor, 1.9 = volume of Arginine solution required to prepare the Sample solution P
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No peaks or shoulders in the chromatogram of the Sample solution that are not present in the chromatogram of the Standard solution are found. Calculate the percentage of single-chain alteplase in the portion of Alteplase taken: Result = (rA/rS) 100 = peak response for single-chain alteplase rA = sum of the responses of all of the alteplase peaks rS Acceptance criteria: NLT 60% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 Acceptance criteria: Meets the requirements for Content Uniformity SPECIFIC TESTS CONSTITUTED SOLUTIONS Acceptance criteria: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. PH 791 Sample solution: Constitute as directed in the labeling. Acceptance criteria: 7.17.5 WATER DETERMINATION, Method I 921: NMT 4.0% BACTERIAL ENDOTOXINS 85: NMT 1 USP Endotoxin Unit/mg STERILITY TESTS 71 Acceptance criteria: Meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. SAFETY Acceptance criteria: Meets the requirements for biologics as set forth for Biological Reactivity Tests, In Vivo 88, Safety TestsBiologicals. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in hermetic, lightresistant containers, and store in a refrigerator. LABELING: Label it to state the biological activity in USP Alteplase Units/vial and the amount of protein/vial. USP REFERENCE STANDARDS 11 USP Alteplase RS USP Endotoxin RS
Acceptance criteria: 95.0%111.0% PROCEDURE 2: PERCENT MONOMER Mobile phase: 34.84 mg/mL of arginine, 158.56 mg/mL of ammonium sulfate, and 100 mL/L of isopropyl alcohol in water. Adjust with phosphoric acid to a pH of 7.3, degas, and pass through a 0.45-m porosity filter. Make adjustments if necessary. System suitability solution: 1.0 mg/mL each of chicken ovalbumin and bovine gamma globulin Standard solution: 1 mg/mL of USP Alteplase RS Sample solution: 1 mg/mL Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 7.5-mm 30-cm; packing L25 Flow rate: 0.51.0 mL/min Injection size: 50 L System suitability Sample: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.6 between gamma globulin and ovalbumin, System suitability solution. Column efficiency: NLT 1200 theoretical plates, Sample solution Analysis Samples: Sample solution and Standard solution Calculate, as a percentage, the monomer in the portion of Alteplase for Injection taken: Result = (rM/rS) 100 = peak response for the alteplase monomer = sum of the responses of all of the alteplase related peaks Acceptance criteria: NLT 95% PROCEDURE 3: SINGLE-CHAIN CONTENT [NOTEWhen constituted with water, Alteplase for Injection meets this requirement.] Mobile phase: 27.6 mg/mL of monobasic sodium phosphate in sodium dodecyl sulfate solution (1 in 1000). Adjust with sodium hydroxide to a pH of 6.8, filter, and degas. Make adjustments if necessary. 0.02 M dithiothreitol solution: 3.12 mg/mL of dithiothreitol in Mobile phase Standard stock solution: 1 mg/mL of USP Alteplase RS Standard solution: Pipet 1 mL of the Standard stock solution into a glass tube. Add 3 mL of 0.02 M dithiothreitol solution, cap the tube, and invert to mix. Heat for 35 min at about 80. Sample stock solution: 1 mg/mL of Alteplase Sample solution: Pipet 1 mL of the Sample stock solution into a glass tube. Add 3 mL of 0.02 M dithiothreitol solution, cap the tube, and invert to mix. Heat for 35 min at about 80. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 7.5-mm 60-cm; packing L25 Flow rate: 0.5 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.1 between the single-chain and twochain alteplase peaks Analysis Samples: Sample solution and Standard solution [NOTEThe major peaks are from single-chain and two-chain alteplase and from higher and lower molecular weight species.] rM rS
Altretamine
(Comment on this Monograph)id=m1678=Altretamine=AMonos.pdf)
210.28 C9H18N6 1,3,5-Triazine-2,4,6-triamine, N,N,N,N,N,N-hexamethyl-; Hexamethylmelamine [645-05-6]. DEFINITION Altretamine contains NLT 98.0% and NMT 102.0% of C9H18N6, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
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ASSAY PROCEDURE Buffer: 790 g/mL of ammonium carbonate; if necessary, adjust to pH of 8.0 0.05 with a solution of formic acid (1 in 10) or ammonium hydroxide (1 in 10). Diluent: Methanol and water (65:35) Mobile phase: Methanol and Buffer (65:35) Standard stock solution: Dissolve 25 mg of USP Altretamine RS in 35 mL of methanol, and dilute to 50 mL with water. Standard solution: 50 g/mL of Altretamine in Diluent, from Standard stock solution Sample stock solution: Dissolve 25 mg of Sample in 35 mL of methanol, and dilute to 50 mL with water. Sample solution: 50 g/mL of Altretamine in Diluent, from Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 227 nm Column: 4.6-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, as a percentage, of C9H18N6 in the portion of Altretamine taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Altretamine RS in Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 40 ppm SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1% rU rS CS
Official Monographs / Altretamine 99

B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: 790 mg/L of ammonium carbonate; if necessary, adjust to pH of 8.0 0.05 with a solution of formic acid (1 in 10) or ammonium hydroxide (1 in 10). Diluent: Methanol and water (65:35) Mobile phase: Methanol and Buffer (65:35), filtered and degassed Standard stock solution: Dissolve 25 mg of USP Altretamine RS in 35 mL of methanol, and dilute to 50 mL with water. Standard solution: 50 g/mL in Diluent from Standard stock solution Sample stock solution: Remove as completely as possible the contents of NLT 20 Capsules, and weigh. Mix the combined contents, and transfer as completely as possible to a 500-mL volumetric flask. Add 325 mL of methanol, and sonicate. Dilute with water to volume. Sample solution: Transfer a volume of the Sample stock solution, equivalent to 10 mg of altretamine, to a 200-mL volumetric flask and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 227 nm Column: 4.6-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Sample: Standard solution and Sample solution Calculate the quantity, as a percentage, of C9H18N6 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Altretamine RS in the Standard solution (mg/mL) CU = nominal concentration of altretamine in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 30 min Detector: UV 242 nm Analysis: Determine the amount of C9H18N6 dissolved from UV absorbances on filtered portions of the solution under test, suitably diluted if necessary with Medium, in comparison with a standard solution having a known concentration of USP Altretamine RS in the same Medium. Tolerances: NLT 80% (Q) of the labeled amount of C9H18N6 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at a controlled room temperature. rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Altretamine RS
Altretamine Capsules
(Comment on this Monograph)id=m1679=Altretamine Capsules=A-Monos.pdf) DEFINITION Altretamine Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of altretamine (C9H18N6). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Remove as completely as possible the contents of 5 Capsules, and dissolve, with shaking, in 10 mL of chloroform. Filter, and evaporate the chloroform solution to dryness.
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USP REFERENCE STANDARDS 11 USP Altretamine RS
Ammonium Alum
(Comment on this Monograph)id=m1680=Ammonium Alum=AMonos.pdf) AINH4(SO4)2 12H2O AINH4(SO4)2 453.33 237.15
Sulfuric acid, aluminum ammonium salt (2:1:1), dodecahydrate; Aluminum ammonium sulfate (1:1:2), dodecahydrate [7784-26-1]. Anhydrous [7784-25-0]. DEFINITION Ammonium Alum contains NLT 99.0% and NMT 100.5% of AlNH4(SO4)2, calculated on the dried basis. IDENTIFICATION A. PROCEDURE Sample solution: 1 in 20 Analysis: Add 1 N sodium hydroxide dropwise to the Sample solution. Acceptance criteria: A precipitate is formed that dissolves in an excess of the reagent with the evolution of ammonia, recognizable by its odor and its alkaline effect upon moistened red litmus paper exposed to the vapor. B. IDENTIFICATION TESTSGENERAL, Aluminum 191 and Sulfate 191: Meets the requirements Sample solution: 1 in 20 ASSAY PROCEDURE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water. Standardize as follows: weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker. Add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V W = weight of aluminum (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrate (mL) Sample: 800 mg Analysis: Transfer the Sample to a 400-mL beaker, moisten with 1 mL of glacial acetic acid, and add 50 mL of water, 50.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS. Warm on a steam bath until solution is complete, and boil gently for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS, and titrate 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 11.86 mg of AlNH4 (SO4)2.
IMPURITIES Inorganic Impurities HEAVY METALS, Method I 231 Sample solution: Dissolve 1 g in water to make 20 mL and add 5 mL of 0.1 N hydrochloric acid. Evaporate the solution in a porcelain evaporating dish to dryness. Treat the residue with 20 mL of water, and add 50 mg of hydroxylamine hydrochloride. Heat the solution on a steam bath for 10 min, cool, and dilute with water to 25 mL. Analysis: Proceed as directed in General Chapter, except to add 50 mg of hydroxylamine hydrochloride to the Standard Preparation. Acceptance criteria: 20 ppm IRON Sample solution: 1 in 150 Analysis: Add 5 drops of potassium ferrocyanide TS to 20 mL of the Sample solution. Acceptance criteria: No blue color is produced immediately. SPECIFIC TESTS LOSS ON DRYING 731 Sample: 2.0 g Analysis: Transfer the Sample, in a tared porcelain crucible, to a muffle furnace at 200. Increase the temperature to 300, and dry at 300 to a constant weight. Cool in a desiccator, and weigh. Acceptance criteria: The sample loses 45.0%48.0% of its weight.
Potassium Alum
(Comment on this Monograph)id=m1685=Potassium Alum=AMonos.pdf) AIK(SO4)2 12H2O AIK(SO4)2 474.39 258.21
Sulfuric acid, aluminum potassium salt (2:1:1), dodecahydrate; Aluminum potassium sulfate (1:1:2) dodecahydrate [7784-24-9]. Anhydrous [10043-67-1]. DEFINITION Potassium Alum contains NLT 99.0% and NMT 100.5% of AlK(SO4)2, calculated on the dried basis. IDENTIFICATION A. PROCEDURE Sample solution: 50 mg/mL in water Analysis: Add 1 N sodium hydroxide dropwise to the Sample solution. Acceptance criteria: A precipitate is formed that dissolves in an excess of the reagent. Ammonia is not evolved (distinction from ammonium alum). B. PROCEDURE Analysis: Hold a sample in a nonluminous flame. Acceptance criteria: A violet color is imparted to the flame. C. PROCEDURE Sample solution: Saturated solution in water Analysis: Add 10 mL of sodium bitartrate TS to 5 mL of the Sample solution. Acceptance criteria: A white, crystalline precipitate is formed within 30 min.
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D. IDENTIFICATION TESTSGENERAL, Aluminum and Sulfate 191 Sample solution: 50 mg/mL in water Acceptance criteria: Meets the requirements ASSAY PROCEDURE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water. Standardize as follows: Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) Ar = atomic weight of aluminum (Al), 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample: 800 mg Analysis: Transfer the Sample to a 400-mL beaker, moisten with 1 mL of glacial acetic acid, and add 50 mL of water, 50.0 mL of Edetate disodium titrant, and 20 mL of acetic acidammonium acetate buffer TS. Warm on a steam bath until solution is complete, and boil gently for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS, and titrate 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 12.91 mg of AlK(SO4)2. Acceptance criteria: 99.0% 100.5% IMPURITIES Inorganic Impurities HEAVY METALS, Method I 231 Sample solution: Dissolve 1 g in water to make 20 mL, and add 5 mL of 0.1 N hydrochloric acid. Evaporate the solution in a porcelain evaporating dish to dryness. Treat the residue with 20 mL of water, and add 50 mg of hydroxylamine hydrochloride. Heat the solution on a steam bath for 10 min, cool, and dilute with water to 25 mL. Analysis: Proceed as directed, except to add 50 mg of hydroxylamine hydrochloride to the Standard Preparation. Acceptance criteria: 20 ppm IRON 241 Sample solution: 1 in 150 Analysis: Add 5 drops of potassium ferrocyanide TS to 20 mL of the Sample solution. Acceptance criteria: No blue color is produced immediately. SPECIFIC TESTS LOSS ON DRYING 731 Sample: 2.0 g Analysis: Transfer the Sample solution in a tared porcelain crucible to a muffle furnace at 200. Increase the temperature to 400, and dry at 400 to constant weight. Cool in a desiccator, and weigh. Acceptance criteria: The sample loses 43.0%46.0% of its weight. W
Official Monographs / Alumina 101

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at room temperature.
Alumina and Magnesia Oral Suspension

(Comment on this Monograph)id=m1700=Alumina and Magnesia Oral Suspension=A-Monos.pdf) DEFINITION Alumina and Magnesia Oral Suspension is a mixture containing aluminum hydroxide [Al(OH)3] and Magnesium Hydroxide [Mg(OH)2]. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium hydroxide [Mg(OH)2]. It may contain a flavoring agent, and may contain suitable antimicrobial agents. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: To a solution of 5 g in 10 mL of 3 N hydrochloric acid add 5 drops of methyl red TS, heat to boiling, add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, then continue boiling for 2 min, and filter. Acceptance criteria: The filtrate meets the requirements. B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the precipitate obtained in Identification A with 20 mg/mL of hot ammonium chloride solution, and dissolve the precipitate in hydrochloric acid. Acceptance criteria: The solution meets the requirements. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water. Standardize as follows: Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution. Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum; 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer a volume of Oral Suspension, previously well shaken in its original container, equivalent to 1200 mg of aluminum hydroxide, to a suitable beaker. Add 20 mL of water, stir, and slowly add 10 mL of hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 10 mL of the Sample solution into a beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant, 20 mL of acetic acidammonium acetate buffer TS, and heat W
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ANC1 A ANC2 M
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= theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq = amount of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of Mg(OH)2, 0.0343 mEq = amount of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg)
near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rosepink. Perform a blank determination, substituting 10 mL of water for the Sample solution. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM HYDROXIDE Sample solution: Proceed as directed under Assay, Aluminum hydroxide. Analysis: Pipet a volume of Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker. Add 200 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol, and mixing). Cool the solution to between 3 and 4 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting 10 mL of water for the Sample solution. Each mL of 0.05 M edetate disodium consumed is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221 Sample solution: Dissolve 5.0 g in the minimum volume of nitric acid required to achieve complete solution, add 1 mL of acid in excess, then add water to make 100 mL, and filter. Acceptance criteria: A 10-mL portion of the Sample solution shows no more chloride than corresponds to 1.0 mL of 0.020 N hydrochloric acid (0.14%). CHLORIDE AND SULFATE, Sulfate 221 Sample solution: Dissolve 5.0 g in 5 mL of 3 N hydrochloric acid, with gentle heating. Cool, add water to make 250 mL, and filter. Acceptance criteria: A 20-mL portion of the Sample solution shows no more sulfate than corresponds to 0.40 mL of 0.020 N sulfuric acid (0.1%). ARSENIC, Method I 211 Standard solution: Prepare as directed in Arsenic 211, except prepare it to contain 5 g of arsenic instead of 3 g. Sample solution: Oral Suspension, equivalent to 0.5 g of Al(OH)3, in 20 mL of 7 N sulfuric acid Acceptance criteria: NMT 10 ppm, based on the Al(OH)3 content HEAVY METALS 231 Sample solution: Oral Suspension, equivalent to 0.24 g of Al(OH)3, in 10 mL of 3 N hydrochloric acid with the aid of heat, filter, if necessary, and dilute with water to 25 mL Acceptance criteria: NMT 83 ppm, based on the Al(OH)3 content SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62 Acceptance criteria: Its total aerobic microbial count does not exceed 100 cfu/mL, and it meets the requirements for absence of Escherichia coli. PH 791: 7.38.5 ACID-NEUTRALIZING CAPACITY 301 Acceptance criteria: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55 (ANC1A) + 0.8 (ANC2M)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing. LABELING: Oral Suspension may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
Alumina and Magnesia Tablets

(Comment on this Monograph)id=m1703=Alumina and Magnesia Tablets=A-Monos.pdf) DEFINITION Alumina and Magnesia Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium hydroxide [Mg(OH)2]. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: To a 0.7-g portion of finely powdered Tablets, add 10 mL of 3 N hydrochloric acid and 5 drops of methyl red TS, heat to boiling, and add 6 N ammonium hydroxide until the color of the solution changes to deep yellow. Continue boiling for 2 min, and filter: the filtrate meets the requirements of the test. B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the precipitate obtained in Identification test A with hot ammonium chloride solution (1 in 50), and dissolve the precipitate in hydrochloric acid: the solution meets the requirements of the test. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV = weight of aluminum in the portion of solution taken (mg) Ar = atomic weight of aluminum, 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 1200 mg of aluminum W
USP 32
hydroxide, to a 150-mL beaker. Add 20 mL of water, stir, and slowly add 30 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 10 mL of Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat near the boiling point for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: Equivalent of 90.0%110.0% of the labeled amounts of Al(OH)3 MAGNESIUM HYDROXIDE Sample solution: Prepare as directed in the Assay for Aluminum Hydroxide. Analysis: Pipet a volume of Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker. Add 200 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black TS in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol). Cool the solution to between 3 and 4 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M edetate disodium consumed is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: Equivalent of 90.0%110.0% of the labeled amounts of Mg(OH)2 PERFORMANCE TESTS DISINTEGRATION 701 Medium: Simulated gastric fluid TS being substituted for water in the test Time: 10 min UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to alumina and to magnesia SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1A) + 0.8(ANC2M) ANC1 A ANC2 M = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq = quantity of Al(OH)3 in the speciment tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of Mg(OH)2, 0.0343 mEq = quantitiy of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg)
Alumina, Magnesia, and Calcium Carbonate Oral Suspension

(Comment on this Monograph)id=m1710=Alumina, Magnesia, and Calcium Carbonate Oral Suspension=A-Monos.pdf) DEFINITION Alumina, Magnesia, and Calcium Carbonate Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amounts of Al(OH)3, Mg(OH)2, and CaCO3. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample solution: To 5 g of Oral Suspension, add 25 mL of 2 N sulfuric acid, stir, and allow to stand for 5 min. Add 25 mL of alcohol, stir, and place in an ice bath for 30 min. Filter while cold. [NOTERetain the filtrate for Identification test B.] Wash the precipitate with 50 mL of 0.75 N sulfuric acid, and discard the washings. Dissolve the precipitate in 3 N hydrochloric acid, and filter. Acceptance criteria: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: To the filtrate obtained in Identification test A, add 5 drops of methyl red TS, and heat to boiling. Add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, continue boiling for 2 min, and filter through hardened filter paper. [NOTERetain the filtrate for Identification test C.] Wash the precipitate with 350 mL of a hot ammonium chloride solution (1 in 50), discarding the washings. Dissolve the precipitate so obtained in 3 N hydrochloric acid. Acceptance criteria: Meets the requirements. C. IDENTIFICATION TESTSGENERAL, Aluminum Magnesium 191 Sample solution: The filtrate obtained in Identification test B Acceptance criteria: Meets the requirements ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution. Calculate the molarity of the solution taken: Result = W/ArV = weight of aluminum in the portion of solution taken (mg) Ar = atomic weight of aluminum, 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer the Oral Suspension, previously well shaken in its original container, equivalent to 600 mg of aluminum hydroxide, to a tared beaker and weigh. Add 20 mL of water, stir, and slowly add 40 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and transfer to a 200-mL volumetric flask. Wash the beaker with W
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Tablets prepared with the use of Dried Aluminum Hydroxide Gel may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
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CHLORIDE AND SULFATE, Sulfate 221 Sample solution: Dissolve 5.0 g in 7 mL of 3 N hydrochloric acid, and gently heat. Cool, add water to make 250 ml, and filter. Acceptance criteria: A 20-mL portion of the Sample solution shows no more sulfate than corresponds to 0.40 mL of 0.020 N sulfuric acid (0.1%). SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62 Acceptance criteria: Its total aerobic microbial count does not exceed 100 cfu/mL, and it meets the requirements of the test for absence of Escherichia coli. PH 791 Acceptance criteria: Between 7.5 and 8.5 ACID-NEUTRALIZING CAPACITY 301 Acceptance criteria: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C) ANC1 A ANC2 M ANC3 C = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq = amount of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of Mg(OH)2, 0.0343 mEq = amount of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of CaCO3, 0.02 mEq = amount of CaCO3 in the specimen tested, based on the labeled quantity (mg)
water, adding the washings to the flask. Add water to volume. Analysis: Pipet 10 mL of Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM HYDROXIDE Sample solution: Transfer Oral Suspension, previously well shaken in its original container, equivalent to 600 mg of aluminum hydroxide, to a tared beaker and weigh. Add 20 mL of water, stir, and slowly add 40 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and transfer to a 200-mL volumetric flask. Wash the beaker with water, adding the washings to the flask. Add water to volume. Analysis: Pipet a volume of Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker, and add 200 mL of water and 20 mL of trolamine, and mix. Add 50 mL of ammoniaammonium chloride buffer TS and 2 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of trolamine and 5 mL of dehydrated alcohol, and mixing). Cool the solution to between 3 and 4 by immersing the beaker in an ice bath, and titrate with 0.05 M edetate disodium VS until the color changes to pure blue. Perform a blank determination, substituting 10 mL of water for the Sample solution. From the volume of 0.05 M edetate disodium consumed, subtract the volume of 0.05 M edetate disodium consumed in the Calcium Carbonate. Each mL of 0.05 M edetate disodium is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% of the labeled amount of magnesium hydroxide [Mg(OH)2] CALCIUM CARBONATE Sample solution: Transfer the Oral Suspension, previously well shaken in its original container, equivalent to 600 mg of aluminum hydroxide, to a tared beaker and weigh. Add 20 mL of water, stir, and slowly add 40 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and transfer to a 200-mL volumetric flask. Wash the beaker with water, adding the washings to the flask. Add water to volume. Analysis: Pipet a volume of the Sample solution, equivalent to 50 mg of calcium carbonate, into a 400-mL beaker, and add 200 mL of water, 5 mL of sodium hydroxide solution (1 in 2), and 250 mg of hydroxy naphthol blue. Stir with a magnetic stirrer, and titrate immediately with 0.05 M edetate disodium VS until the solution is distinctly blue. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. Acceptance criteria: NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3) IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221 Sample solution: Dissolve 5.0 g 3 mL of nitric acid, add water to make 100 mL, and filter. Acceptance criteria: A 10-mL portion of the Sample solution shows no more chloride than corresponds to 1.0 mL of 0.020 N hydrochloric acid (0.14%).
OTHER REQUIREMENTS ARSENIC, Method I 211 Sample solution: Gel, equivalent to 0.5 g of Al(OH)3, in 20 mL of 7 N sulfuric acid Standard solution: Prepare as directed in the test for Arsenic 211, except to prepare it to contain 5 g of arsenic instead of 3 g. Acceptance criteria: NMT 10 ppm, based on the Al(OH)3 content HEAVY METALS 231 Sample solution: Gel, equivalent to 0.24 g of Al(OH)3, in 10 mL of 3 N hydrochloric acid with the aid of heat, filter if necessary, and dilute with water to 25 mL. Acceptance criteria: NMT 83 ppm, based on the Al(OH)3 content ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing. LABELING: Oral Suspension may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
Alumina, Magnesia, and Calcium Carbonate Tablets

(Comment on this Monograph)id=m1713a=Alumina, Magnesia, and Calcium Carbonate Tablets=A-Monos.pdf) (Current titlenot to change until February 1, 2010) Monograph title changeto become official February 1, 2010 See Alumina, Magnesia, and Calcium Carbonate Chewable Tablets
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DEFINITION Alumina, Magnesia, and Calcium Carbonate Chewable Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3], magnesium hydroxide [Mg(OH)2], and calcium carbonate (CaCO3). IDENTIFICATION A. PROCEDURE Sample solution: To 3 g of finely powdered Chewable Tablets, add 25 mL of water and 25 mL of 2 N sulfuric acid, stir, and heat on a steam bath for 10 min. Cool, add 50 mL of alcohol, and stir. Analysis 1: Place the mixture obtained above in an ice bath for 30 min. Filter while cold, retaining the filtrate for Identification test B. Wash the precipitate with 50 mL of 0.75 N sulfuric acid, and discard the washings: the precipitate so obtained, dissolved in 3 N hydrochloric acid and filtered, meets the requirements for Identification TestsGeneral, Calcium 191. B. PROCEDURE Analysis 2: To the filtrate obtained in Identification test A add 5 drops of methyl red TS, and heat to boiling. Add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, continue boiling for 2 min, and filter through hardened filter paper. [NOTERetain the filtrate for Identification test C.] Wash the precipitate with 350 mL of a hot ammonium chloride solution (1 in 50), discarding the washings: the precipitate so obtained, dissolved in 3 N hydrochloric acid meets the requirements for Identification TestsGeneral, Aluminum 191. C. PROCEDURE Analysis 3: The filtrate obtained in Identification test B meets the requirements for Identification TestsGeneral, Magnesium 191. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 g of edetate disodium in water to make 1000 mL Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Weigh and finely powder NLT 20 Chewable Tablets. Transfer a portion of the powder, equivalent to 600 mg of aluminum hydroxide, to a beaker, add 20 mL of water, and slowly add 40 mL of 3 N hydrochloric acid, with mixing. Heat the mixture to boiling, cool, and filter into a 200-mL volumetric flask. Wash the beaker with water, adding the washings to the filter. Add water to volume. Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named W

and with continuous stirring, 25.0 mL of 0.05 M Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS, and mix. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% of the labeled amount of aluminum hydroxide [Al(OH)3] MAGNESIUM HYDROXIDE Edetate disodium titrant and Sample solution: Prepare as directed in the Assay for Aluminum Hydroxide. Analysis: Pipet a volume of Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker, add 200 mL of water and 20 mL of trolamine, and mix. Add 50 mL of ammoniaammonium chloride buffer TS and 2 drops of eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of trolamine and 5 mL of dehydrated alcohol, and mixing). Cool the solution to 34 by immersing the beaker in an ice bath, and titrate with 0.05 M edetate disodium VS until the color changes to pure blue. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. From the volume of 0.05 M edetate disodium consumed, subtract the volume of 0.05 M edetate disodium consumed in the Assay for Calcium Carbonate. Each mL of 0.05 M edetate disodium is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% of the labeled amount of magnesium hydroxide [Mg(OH)2] CALCIUM CARBONATE Edetate disodium titrant and Sample solution: Prepare as directed in the Assay for Aluminum Hydroxide. Analysis: Pipet a volume of the Sample solution, equivalent to 50 mg of calcium carbonate, into a 400-mL beaker, and add 200 mL of water, 5 mL of sodium hydroxide solution (1 in 2), and 250 mg of hydroxy naphthol blue. Stir with a magnetic stirrer, and titrate immediately with 0.05 M edetate disodium VS until the solution is distinctly blue. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of calcium carbonate (CaCO3). Acceptance criteria: 90.0%110.0% of the labeled amount of calcium carbonate (CaCO3) PERFORMANCE TESTS DISINTEGRATION 701 Time: 45 min UNIFORMITY OF DOSAGE UNITS 905 Acceptance criteria: Meets the requirements for Weight Variation with respect to alumina, to magnesia, and to calcium carbonate SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Acceptance criteria: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C) ANC1 A ANC2 M = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq = amount of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of Mg(OH)2, 0.0343 mEq = amount of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg)
106
ANC3 C

= theoretical acid-neutralizing capacity of CaCO3, 0.02 mEq = amount of CaCO3 in the specimen tested, based on the labeled quantity (mg)
USP 32
solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Weigh and finely powder NLT 20 Chewable Tablets. Transfer a portion of the powder, equivalent to 600 mg of aluminum hydroxide, to a beaker, add 20 mL of water, and slowly add 40 mL of 3 N hydrochloric acid, with mixing. Heat the mixture to boiling, cool, and filter into a 200-mL volumetric flask. Wash the beaker with water, adding the washings to the filter. Add water to volume. Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of 0.05 M Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS, and mix. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% of the labeled amount of aluminum hydroxide [Al(OH)3] MAGNESIUM HYDROXIDE Edetate disodium titrant and Sample solution: Prepare as directed in the Assay for Aluminum Hydroxide. Analysis: Pipet a volume of Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker, add 200 mL of water and 20 mL of trolamine, and mix. Add 50 mL of ammoniaammonium chloride buffer TS and 2 drops of eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of trolamine and 5 mL of dehydrated alcohol, and mixing). Cool the solution to 3 to 4 by immersing the beaker in an ice bath, and titrate with 0.05 M edetate disodium VS until the color changes to pure blue. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. From the volume of 0.05 M edetate disodium consumed, subtract the volume of 0.05 M edetate disodium consumed in the Assay for Calcium Carbonate. Each mL of 0.05 M edetate disodium is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% of the labeled amount of magnesium hydroxide [Mg(OH)2] CALCIUM CARBONATE Edetate disodium titrant and Sample solution: Prepare as directed in the Assay for Aluminum Hydroxide. Analysis: Pipet a volume of the Sample solution, equivalent to 50 mg of calcium carbonate, into a 400-mL beaker, and add 200 mL of water, 5 mL of sodium hydroxide solution (1 in 2), and 250 mg of hydroxy naphthol blue. Stir with a magnetic stirrer, and titrate immediately with 0.05 M edetate disodium VS until the solution is distinctly blue. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of calcium carbonate (CaCO3). W
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Chewable Tablets to indicate that they are to be chewed before being swallowed. Chewable Tablets prepared using dried aluminum hydroxide gel may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
Alumina, Magnesia, and Calcium Carbonate Chewable Tablets

(Comment on this Monograph)id=m1713b=Alumina, Magnesia, and Calcium Carbonate Chewable Tablets=A-Monos.pdf) (Monograph under this new titleto become official February 1, 2010) Current monograph title is Alumina, Magnesia, and Calcium Carbonate Tablets DEFINITION Alumina, Magnesia, and Calcium Carbonate Chewable Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3], magnesium hydroxide [Mg(OH)2], and calcium carbonate (CaCO3). IDENTIFICATION Sample solution: To 3 g of finely powdered Chewable Tablets, add 25 mL of water and 25 mL of 2 N sulfuric acid, stir, and heat on a steam bath for 10 min. Cool, add 50 mL of alcohol, and stir: the mixture so obtained meets the following requirements: A. PROCEDURE Analysis 1: Place the mixture obtained above in an ice bath for 30 min. Filter while cold retaining the filtrate for Identification test B. Wash the precipitate with 50 mL of 0.75 N sulfuric acid, and discard the washings: the precipitate so obtained, dissolved in 3 N hydrochloric acid and filtered, meets the requirements for Identification TestsGeneral, Calcium 191. B. PROCEDURE Analysis 2: To the filtrate obtained in Identification test A add 5 drops of methyl red TS, and heat to boiling. Add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, continue boiling for 2 min, and filter through hardened filter paper. [NOTERetain the filtrate for Identification test C.] Wash the precipitate with 350 mL of a hot ammonium chloride solution (1 in 50), discarding the washings: the precipitate so obtained, dissolved in 3 N hydrochloric acid meets the requirements for Identification TestsGeneral, Aluminum 191. C. PROCEDURE Analysis 3: The filtrate obtained in Identification test B meets the requirements for Identification TestsGeneral, Magnesium 191. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 g of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this
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Acceptance criteria: 90.0%110.0% of the labeled amount of calcium carbonate (CaCO3) PERFORMANCE TESTS DISINTEGRATION 701 Time: 45 min UNIFORMITY OF DOSAGE UNITS 905 Acceptance criteria: Meets the requirements for Weight Variation with respect to alumina, to magnesia, and to calcium carbonate. SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Acceptance criteria: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C) ANC1 A ANC2 M ANC3 C = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq = amount of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of Mg(OH)2, 0.0343 mEq = amount of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of CaCO3, 0.02 mEq = amount of CaCO3 in the specimen tested, based on the labeled quantity (mg)

Acceptance criteria: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: To the filtrate obtained in Identification test A, add 5 drops of methyl red TS, and heat to boiling. Add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, continue boiling for 2 min, and filter through hardened filter paper. [NOTERetain the filtrate for Identification test C.] Wash the precipitate with 350 mL of a hot ammonium chloride solution (1 in 50), discarding the washings. Dissolve the precipitate so obtained in 3 N hydrochloric acid. Acceptance criteria: Meets the requirements C. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: The filtrate obtained in Identification test B Acceptance criteria: Meets the requirements D. INFRARED ABSORPTION 197S Sample solution: Transfer the equivalent to 60 mg of simethicone from Chewable Tablets (weigh and finely powder NLT 20 Chewable Tablets), to a suitable screwcapped bottle. Add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid (see the Assay for Dimethylpolysiloxane), and allow to stand, with frequent shaking, until the Chewable Tablets are dissolved. Transfer the contents of the bottle to a separator, shake, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Sample solution. Analysis: Proceed as directed using a 0.5-mm cell. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) V = volume of Edetate disodium titrant consumed (mL) Ar = atomic weight of aluminum, 26.98 Sample solution: Transfer the equivalent to 665 mg of aluminum hydroxide from a number of Chewable Tablets, to a suitable beaker. Add 15 mL of hydrochloric acid, and swirl to dissolve the Chewable Tablets. Add 80 mL of water and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 20 mL of Sample solution into a 250-mL beaker, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the W
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Chewable Tablets to indicate that they are to be chewed before being swallowed. Chewable Tablets prepared using dried aluminum hydroxide gel may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
Alumina, Magnesia, Calcium Carbonate, and Simethicone Tablets

(Comment on this Monograph)id=m1720=Alumina, Magnesia, Calcium Carbonate, and Simethicone Tablets=A-Monos.pdf) (Current titlenot to change until February 1, 2010) Monograph title changeto become official February 1, 2010 See Alumina, Magnesia, Calcium Carbonate, and Simethicone Chewable Tablets. DEFINITION Alumina, Magnesia, Calcium Carbonate, and Simethicone Chewable Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3], magnesium hydroxide [Mg(OH)2], and calcium carbonate (CaCO3), and an amount of polydimethylsiloxane ([(CH3)2 SiO]n) that is NLT 85.0% and NMT 115.0% of the labeled amount of simethicone. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample solution: Cut a Chewable Tablet into pieces, add 50 mL of 1 N sulfuric acid, stir until the pieces disintegrate, and heat on a steam bath for 10 min. Cool, add 50 mL of alcohol, and stir. Place in an ice bath for 30 min. Filter while cold. [NOTERetain the filtrate for Identification test B.] Wash the precipitate with 50 mL of 0.75 N sulfuric acid, and discard the washings. Dissolve the precipitate so obtained in 3 N hydrochloric acid and filter.
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Acceptance criteria: 90.0%110.0% CALCIUM CARBONATE Sample solution: Transfer the equivalent to 665 mg of aluminum hydroxide from a number of Chewable Tablets, to a suitable beaker. Add 15 mL of hydrochloric acid, and swirl to dissolve the Chewable Tablets. Add 80 mL of water and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet a volume of the Sample solution, equivalent to 50 mg of calcium carbonate, into a 400-mL beaker, and add 200 mL of water, a volume of sodium hydroxide solution (1 in 2) equivalent to the volume of the Sample solution taken, and 250 mg of hydroxy naphthol blue. Stir with a magnetic stirrer, and titrate immediately with 0.05 M edetate disodium VS until the solution is distinctly blue. Perform a blank determination, substituting a volume of water equivalent to the volume of the Sample solution taken, and make any necessary correction. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. Acceptance criteria: 90.0%110.0% POLYDIMETHYLSILOXANE Dilute hydrochloric acid: 400 mL of hydrochloric acid with sufficient water to make 1000 mL Standard solution: Transfer 60 mg of USP Polydimethylsiloxane RS to a separator, add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid, shake for 30 s, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Standard solution. Sample solution: Transfer the equivalent to 60 mg of simethicone from Chewable Tablets (weigh and finely powder NLT 20 Chewable Tablets), to a suitable screwcapped bottle. Add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid, and allow to stand, with frequent shaking, until the Chewable Tablets are dissolved. Transfer the contents of the bottle to a separator, shake, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Sample solution. Blank: Place 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid in a separator, shake for 30 s, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Blank. Analysis: Concomitantly determine the absorbances of the Standard solution and the Sample solution in 0.5-mm cells at the wavelength of maximum absorbance at 7.9 m, with a suitable IR spectrophotometer, using the Blank to set the instrument. Calculate the percentage of [(CH3)2 SiO]n in each Tablet taken: Result = (AU/AS) (CS/CU) 100 AU AS CS = absorbance of the Sample solution = absorbance of the Standard solution = concentration of polydimethylsiloxane in the Standard solution (mg/mL)
solution near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from greenviolet to rose-pink. Perform a blank determination, substituting 20 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM HYDROXIDE Lanthanum chloride solution: Transfer 17.6 g of lanthanum chloride to a 200-mL volumetric flask, add 100 mL of water, and carefully add 50 mL of hydrochloric acid. Allow to cool, and dilute with water to volume. Dilute hydrochloric acid: Dilute 226 mL of hydrochloric acid with water to 1000 mL. Potassium chloride solution: 30 mg/mL Magnesium stock solution: Transfer 1.000 g of magnesium metal to a 1000-mL volumetric flask containing 10 mL of water, slowly add 10 mL of hydrochloric acid, and swirl to dissolve the metal. Dilute with water to volume. Magnesium solution: 20 g/mL of magnesium (Mg) in water, from Magnesium stock solution Standard solutions: To three separate, 100-mL volumetric flasks each containing 5.0 mL of Lanthanum chloride solution, add 1.0, 2.0, and 3.0 mL, respectively, of the Magnesium solution. Dilute each with water to volume. These solutions contain 0.1, 0.2, and 0.3 g/mL of magnesium (Mg), respectively. Sample stock solution: Transfer the equivalent to 250 mg of magnesium hydroxide (100 mg of magnesium) from a number of Chewable Tablets, to a 1000-mL volumetric flask. Add 500 mL of Dilute hydrochloric acid, and swirl to disintegrate the Chewable Tablets. Add 100.0 mL of Potassium chloride solution, and dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Sample solution: Transfer 2.0 mL of Sample stock solution to a second 100-mL volumetric flask, add 5.0 mL of Lanthanum chloride solution, and dilute with water to volume. Blank: Add 50 mL of Dilute hydrochloric acid and 10.0 mL of Potassium chloride solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 10.0 mL of this solution to a second 100-mL volumetric flask, and dilute to volume with water. Transfer 2.0 mL of this solution to a third, 100-mL volumetric flask, add 5.0 mL of Lanthanum chloride solution, and dilute with water to volume. Analysis: Concomitantly determine the absorbances of the Standard solutions and the Sample solution at the magnesium emission line at 285.2 nm, with a suitable atomic absorption spectrophotometer (see Spectrophotometry and LightScattering 851) equipped with a magnesium hollowcathode lamp and an airacetylene flame, using the Blank to set the instrument. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of magnesium, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of magnesium in the Sample solution. Calculate the percentage of Mg(OH)2 in each Tablet taken: Result = (Mr/Ar) (500C/N) 100 Mr Ar C N = molecular weight of magnesium hydroxide, 58.34 = atomic weight of magnesium, 24.305 = nominal concentration of magnesium in the Sample solution (g/mL) = number of Chewable Tablets taken to prepare the Sample solution
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= nominal concentration of polydimethylsiloxane in the Sample solution (mg/mL) Acceptance criteria: 85.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for Weight Variation with respect to aluminum hydroxide, to magnesium hydroxide, and to calcium carbonate SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: The total aerobic microbial count is NMT 200 cfu/g; the total combined molds and yeasts count is NMT 200 cfu/g; and the Chewable Tablets meet the requirements of the test for the absence of Salmonella species and Escherichia coli. ACID-NEUTRALIZING CAPACITY 301 Sample solution: Equivalent to 120 mEq of acidneutralizing capacity from a counted number of Chewable Tablets, in 400 mL of water Transfer the mixture to a 500-mL volumetric flask, and dilute with water to volume. Analysis: Proceed as directed in the section Procedure for Powders, Effervescent Solids, Suspensions and Other Liquids, Nonchewable Chewable Tablets, Chewable Chewable Tablets, and Capsules using 75.0 mL of the Sample solution. Acceptance criteria: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C) = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq A = quantity of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of Mg(OH)2, ANC2 0.0343 mEq M = quantity of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of CaCO3, ANC3 0.02 mEq C = quantity of CaCO3 in the specimen tested, based on the labeled quantity (mg) DEFOAMING ACTIVITY Foaming solution: 10 mg/mL of octoxynol 9 in 0.3 N hydrochloric acid Sample: Equivalent to 20 mg of simethicone from an amount of Chewable Tablets, cut into small pieces and mixed [NOTEWeigh NLT 10 Chewable Tablets.] Analysis: [NOTEFor each test, use a clean, 250-mL cylindrical glass jar.] Transfer the Sample to a clean, 250-mL cylindrical glass jar, fitted with a 50-mm cap, containing 100 mL of Foaming solution that has been warmed to 37. After effervescence ceases, cap the jar, and clamp it in an upright position on a wrist-action shaker. Using a radius of 13.3 0.4 cm (measured from the center of shaft to the center of the bottle), shake for 10 s through an arc of 10 at a frequency of 300 30 strokes/min. Record the time required for the foam to collapse. The time, in seconds, for foam collapse is determined at the instant the first portion of foam-free liquid surface appears, measured from the end of the shaking period. Acceptance criteria: The defoaming activity time does not exceed 45 s. SODIUM CONTENT Potassium chloride solution: 30 mg/mL of potassium chloride Dilute hydrochloric acid: Dilute 226 mL of hydrochloric acid with sufficient water to make 1000 mL. ANC1 CU

Standard stock solution: Transfer 2.5420 g of sodium chloride, previously dried at 105 for 2 h, to a 1000-mL volumetric flask, dissolve in and dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 10.0 mL of this solution to a second 100-mL volumetric flask, and dilute with water to volume. Standard solutions: To three separate, 100-mL volumetric flasks, each containing 10.0 mL of Potassium chloride solution and 3.0 mL of Dilute hydrochloric acid, add 10.0, 20.0, and 30.0 mL, respectively, of the Standard stock solution. The resulting Standard solutions contain 1.0, 2.0, and 3.0 g/mL of sodium, respectively. Sample stock solution: Weigh 10 Chewable Tablets, and determine the average weight, A, in mg. Cut 4 Chewable Tablets into pieces, combine the pieces, and weigh them. Transfer the combined pieces to a 500-mL volumetric flask, add 150 mL of Dilute hydrochloric acid, and swirl gently to dissolve the pieces. Dilute with water to volume. Sample solution: Transfer 10.0 mL of the Sample stock solution to a 100-mL volumetric flask, add 10.0 mL of Potassium chloride solution, and dilute with water to volume. Blank solution: Combine 3.0 mL of Dilute hydrochloric acid and 10.0 mL of Potassium chloride solution in a 100-mL volumetric flask, and dilute with water to volume. Analysis: Concomitantly determine the absorbances of the Standard solutions and the Sample solution at the sodium emission line at 589.0 nm with a suitable atomic absorption spectrophotometer (see Spectrophotometry and LightScattering 851) equipped with a sodium hollow-cathode lamp and an airacetylene flame, using the Blank. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of sodium, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of sodium in the Sample solution. Calculate the quantity of Na, in mg, in each Chewable Tablet taken: Result = (A/W) C F = average weight of each tablet (mg) = weight of the portion of Chewable Tablets taken to prepare the Sample solution (mg) C = concentration of sodium in the Sample solution (g/mL) F = correction factor, 5 Acceptance criteria: Chewable Tablets contain NMT 5 mg/Tablet of sodium, except when labeled as containing more than 5 mg/Tablet of sodium; then they contain NMT 110% of the labeled amount. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The labeling indicates that the Chewable Tablets are to be chewed before swallowing. Label the Chewable Tablets to state the sodium content, if it is greater than 5 mg/Chewable Tablet. USP REFERENCE STANDARDS 11 USP Polydimethylsiloxane RS A W
Alumina, Magnesia, Calcium Carbonate, and Simethicone Chewable Tablets

(Comment on this Monograph)id=m1720=Alumina, Magnesia, Calcium Carbonate, and Simethicone Chewable Tablets=AMonos.pdf) (Monograph under this new titleto become official February 1, 2010)
110

Calculate the molarity of the solution taken: Result = W/Ar V
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(Current monograph title is Alumina, Magnesia, Calcium Carbonate, and Simethicone Tablets.) DEFINITION Alumina, Magnesia, Calcium Carbonate, and Simethicone Chewable Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3], magnesium hydroxide [Mg(OH)2], and calcium carbonate (CaCO3), and an amount of polydimethylsiloxane ([(CH3)2 SiO]n) that is NLT 85.0% and NMT 115.0% of the labeled amount of simethicone. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample solution: Cut a Chewable Tablet into pieces, add 50 mL of 1 N sulfuric acid, stir until the pieces disintegrate, and heat on a steam bath for 10 min. Cool, add 50 mL of alcohol, and stir. Place in an ice bath for 30 min. Filter while cold. [NOTERetain the filtrate for Identification test B.] Wash the precipitate with 50 mL of 0.75 N sulfuric acid, and discard the washings. Dissolve the precipitate so obtained in 3 N hydrochloric acid, and filter. Acceptance criteria: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: To the filtrate obtained in Identification test A, add 5 drops of methyl red TS, and heat to boiling. Add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, continue boiling for 2 min, and filter through hardened filter paper. [NOTERetain the filtrate for Identification test C.] Wash the precipitate with 350 mL of a hot ammonium chloride solution (1 in 50), discarding the washings. Dissolve the precipitate so obtained in 3 N hydrochloric acid. Acceptance criteria: Meets the requirements C. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: The filtrate obtained in Identification test B Acceptance criteria: Meets the requirements D. INFRARED ABSORPTION 197S Sample solution: Transfer the equivalent to 60 mg of simethicone from Chewable Tablets (weigh and finely powder NLT 20 Chewable Tablets), to a suitable screwcapped bottle, add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid (see the Assay for Dimethylpolysiloxane), and allow to stand, with frequent shaking, until the Chewable Tablets are dissolved. Transfer the contents of the bottle to a separator, shake, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Sample solution. Analysis: Proceed as directed using a 0.5-mm cell. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction.
= weight of aluminum in the portion of solution taken (mg) V = volume of Edetate disodium titrant consumed (mL) Ar = atomic weight of aluminum, 26.98 Sample solution: Transfer the equivalent to 665 mg of aluminum hydroxide from a counted number of Chewable Tablets, to a suitable beaker. Add 15 mL of hydrochloric acid, and swirl to dissolve the Chewable Tablets. Add 80 mL of water and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 20 mL of Sample solution into a 250-mL beaker, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from greenviolet to rose-pink. Perform a blank determination, substituting 20 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM HYDROXIDE Lanthanum chloride solution: Transfer 17.6 g of lanthanum chloride to a 200-mL volumetric flask, add 100 mL of water, and carefully add 50 mL of hydrochloric acid. Allow to cool, and dilute with water to volume. Dilute hydrochloric acid: Dilute 226 mL of hydrochloric acid to 1000 mL with water. Potassium chloride solution: 30 mg/mL Magnesium stock solution: Transfer 1.000 g of magnesium metal to a 1000-mL volumetric flask containing 10 mL of water, slowly add 10 mL of hydrochloric acid, and swirl to dissolve the metal. Dilute with water to volume. Magnesium solution: 20 g/mL of magnesium (Mg) in water, from Magnesium stock solution Standard solutions: To three separate, 100-mL volumetric flasks each containing 5.0 mL of Lanthanum chloride solution, add 1.0, 2.0, and 3.0 mL, respectively, of the Magnesium solution. Dilute each with water to volume. These solutions contain 0.1, 0.2, and 0.3 g/mL of Mg, respectively. Sample stock solution: Transfer the equivalent to 250 mg of magnesium hydroxide (100 mg of magnesium) from a number of Chewable Tablets, to a 1000-mL volumetric flask. Add 500 mL of Dilute hydrochloric acid, and swirl to dissolve the Chewable Tablets. Add 100.0 mL of Potassium chloride solution, and dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Sample solution: Transfer 2.0 mL of Sample stock solution to a second 100-mL volumetric flask, add 5.0 mL of Lanthanum chloride solution, and dilute with water to volume. Blank: Add 50 mL of Dilute hydrochloric acid and 10.0 mL of Potassium chloride solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 10.0 mL of this solution to a second 100-mL volumetric flask, and dilute with water to volume. Transfer 2.0 mL of this solution to a third 100-mL volumetric flask, add 5.0 mL of Lanthanum chloride solution, and dilute with water to volume. Analysis: Concomitantly determine the absorbances of the Standard solutions and the Sample solution at the magnesium emission line at 285.2 nm, with a suitable atomic absorption spectrophotometer (see Spectrophotometry and Light-
USP 32
Scattering 851) equipped with a magnesium hollowcathode lamp and an airacetylene flame, using the Blank to set the instrument. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of magnesium, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of magnesium in the Sample solution. Calculate the percentage of Mg(OH)2 in each Tablet taken: Result = (Mr/Ar) (500C/N) 100 = molecular weight of magnesium hydroxide, 58.34 Ar = atomic weight of magnesium, 24.305 C = nominal concentration of magnesium in the Sample solution (g/mL) N = number of Chewable Tablets taken to prepare the Sample solution Acceptance criteria: 90.0%110.0% CALCIUM CARBONATE Sample solution: Transfer the equivalent to 665 mg of aluminum hydroxide from a counted number of Chewable Tablets, to a suitable beaker. Add 15 mL of hydrochloric acid, and swirl to dissolve the Chewable Tablets. Add 80 mL of water and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet a volume of the Sample solution, equivalent to 50 mg of calcium carbonate, into a 400-mL beaker, and add 200 mL of water, a volume of sodium hydroxide solution (1 in 2) equivalent to the volume of the Sample solution taken, and 250 mg of hydroxy naphthol blue. Stir with a magnetic stirrer, and titrate immediately with 0.05 M edetate disodium VS until the solution is distinctly blue. Perform a blank determination, substituting a volume of water equivalent to the volume of the Sample solution taken, and make any necessary correction. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of calcium carbonate (CaCO3). Acceptance criteria: 90.0%110.0% POLYDIMETHYLSILOXANE Dilute hydrochloric acid: Dilute 400 mL of hydrochloric acid with sufficient water to make 1000 mL. Standard solution: Transfer 60 mg of USP Polydimethylsiloxane RS to a separator, add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid, shake for 30 s, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Standard solution. Sample solution: Transfer the equivalent to 60 mg of simethicone from Chewable Tablets (weigh and finely powder NLT 20 Chewable Tablets), to a suitable screwcapped bottle. Add 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid, and allow to stand, with frequent shaking, until the Chewable Tablets are dissolved. Transfer the contents of the bottle to a separator, shake, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screwcap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Sample solution. Blank: Place 30.0 mL of chloroform and 60 mL of Dilute hydrochloric acid in a separator, shake for 30 s, and allow the phases to separate. Remove 10 mL of the lower, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap Mr

that has an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The supernatant so obtained is the Blank. Analysis: Concomitantly determine the absorbances of the Standard solution and the Sample solution in 0.5-mm cells at the wavelength of maximum absorbance at 7.9 m, with a suitable IR spectrophotometer, using the Blank to set the instrument. Calculate the percentage of [(CH3)2 SiO]n in each Tablet taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of polydimethylsiloxane in the Standard solution (mg/mL) = nominal concentration of polydimethylsiloxane in CU the Sample solution (mg/mL) Acceptance criteria: 85.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 Meet the requirements for Weight Variation with respect to aluminum hydroxide, to magnesium hydroxide, and to calcium carbonate. SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: The total aerobic microbial count is NMT 200 cfu/g; the total combined molds and yeasts count is NMT 200 cfu/g; and the Chewable Tablets meet the requirements of the test for the absence of Salmonella species and Escherichia coli. ACID-NEUTRALIZING CAPACITY 301 Sample solution: Equivalent to 120 mEq of acidneutralizing capacity from a counted number of Chewable Tablets, in 400 mL of water. Transfer the mixture to a 500mL volumetric flask, and dilute with water to volume. Analysis: Proceed as directed in the section Procedure for Powders, Effervescent Solids, Suspensions and Other Liquids, Nonchewable Chewable Tablets, Chewable Chewable Tablets, and Capsules using 75.0 mL of the Sample solution. Acceptance criteria: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) + 0.9(ANC3 C) = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq A = quantity of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of Mg(OH)2, ANC2 0.0343 mEq M = quantity of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg) = theoretical acid-neutralizing capacity of CaCO3, ANC3 0.02 mEq C = quantity of CaCO3 in the specimen tested, based on the labeled quantity (mg) DEFOAMING ACTIVITY Foaming solution: 10 mg/mL of octoxynol 9 in 0.3 N hydrochloric acid Sample: Equivalent to 20 mg of simethicone from an amount of Chewable Tablets, cut into small pieces and mixed [NOTEWeigh NLT 10 Chewable Tablets.] Analysis: [NOTEFor each test, use a clean, 250-mL cylindrical glass jar.] Transfer the Sample to a clean, 250-mL cylindrical glass jar, fitted with a 50-mm cap, containing 100 mL of Foaming solution that has been warmed to 37. After effervescence ceases, cap the jar, and clamp it in an ANC1 AU AS CS
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Tablets to state the sodium content, if it is greater than 5 mg/Chewable Tablet. USP REFERENCE STANDARDS 11 USP Polydimethylsiloxane RS
upright position on a wrist-action shaker. Using a radius of 13.3 0.4 cm (measured from the center of the shaft to the center of the bottle), shake for 10 s through an arc of 10 at a frequency of 300 30 strokes/min. Record the time required for the foam to collapse. The time, in seconds, for foam collapse is determined at the instant the first portion of foam-free liquid surface appears, measured from the end of the shaking period. Acceptance criteria: The defoaming activity time does not exceed 45 s. SODIUM CONTENT Potassium chloride solution: 30 mg/mL of potassium chloride Dilute hydrochloric acid: Dilute 226 mL of hydrochloric acid with sufficient water to make 1000 mL. Standard stock solution: Transfer 2.5420 g of sodium chloride, previously dried at 105 for 2 h, to a 1000-mL volumetric flask, and dissolve in and dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 10.0 mL of this solution to a second 100-mL volumetric flask, and dilute with water to volume. Standard solutions: To three separate 100-mL volumetric flasks, each containing 10.0 mL of Potassium chloride solution and 3.0 mL of Dilute hydrochloric acid, add 10.0, 20.0, and 30.0 mL, respectively, of the Standard stock solution. The resulting Standard solutions contain 1.0, 2.0, and 3.0 g/mL of sodium (Na), respectively. Sample stock solution: Weigh 10 Chewable Tablets, and determine the average weight, A, in mg. Cut 4 Chewable Tablets into pieces, combine the pieces, and weigh them. Transfer the combined pieces to a 500-mL volumetric flask, add 150 mL of Dilute hydrochloric acid, and swirl gently to dissolve the pieces. Dilute with water to volume. Sample solution: Transfer 10.0 mL of the Sample stock solution to a 100-mL volumetric flask, add 10.0 mL of Potassium chloride solution, and dilute with water to volume. Blank solution: Combine 3.0 mL of Dilute hydrochloric acid and 10.0 mL of Potassium chloride solution in a 100-mL volumetric flask, and dilute with water to volume. Analysis: Concomitantly determine the absorbances of the Standard solutions and the Sample solution at the sodium emission line at 589.0 nm with a suitable atomic absorption spectrophotometer (see Spectrophotometry and LightScattering 851) equipped with a sodium hollowcathode lamp and an airacetylene flame, using the Blank solution as the blank. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of sodium, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of sodium in the Sample solution. Calculate the quantity, in mg, of Na in each Chewable Tablet taken: Result = (A/W) C F = average weight of each Tablet (mg) = weight of the portion of Chewable Tablets taken to prepare the Sample solution (mg) C = concentration of sodium in the Sample solution (g/mL) F = correction factor, 5 Acceptance criteria: Chewable Tablets contain NMT 5 mg/Tablet of sodium, except when labeled as containing more than 5 mg/Tablet of sodium; then they contain NMT 110% of the labeled amount. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The labeling indicates that the Chewable Tablets are to be chewed before swallowing. Label the Chewable A W
Alumina, Magnesia, and Simethicone Oral Suspension

(Comment on this Monograph)id=m1750=Alumina, Magnesia, and Simethicone Oral Suspension=A-Monos.pdf) DEFINITION Alumina, Magnesia, and Simethicone Oral Suspension contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane [(CH3)2 SiO]n that is NLT 85.0% and NMT 115.0% of the labeled amount of simethicone. IDENTIFICATION A. INFRARED ABSORPTION 197S Sample solution: Transfer an amount of Oral Suspension equivalent to 50 mg of simethicone to a suitable round, narrow-mouth, screw-capped, 120-mL bottle, add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 25.0 mL of toluene, close the bottle securely with a cap having an inert liner, and shake for 15 min on a reciprocating shaker (e.g., about 200 oscillations per minute and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator. Remove about 5 mL of the upper, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The clear supernatant is the Sample solution. Analysis: Proceed as directed using a 0.5-mm cell. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Add 5 g of sample to 10 mL of 3 N hydrochloric acid, then add 5 drops of methyl red TS, heat to boiling, add 6 N ammonium hydroxide until the color of the solution just changes to deep yellow, then continue boiling for 2 min and filter. The filtrate so obtained is the Sample solution. Acceptance criteria: Meets the requirements C. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the precipitate from Identification test B with hot ammonium chloride solution (1 in 50), and dissolve the precipitate in hydrochloric acid. Divide this solution into two portions. Analysis 1: Add, dropwise, 6 N ammonium hydroxide to one portion of the Sample solution. Acceptance criteria 1: Yields a gelatinous white precipitate, which does not dissolve in an excess of 6 N ammonium hydroxide. Analysis 2: Add, dropwise, 1 N sodium hydroxide to the second portion of the Sample solution. Acceptance criteria 2: Yields a gelatinous white precipitate, which dissolves in an excess of 1 N sodium hydroxide, leaving some turbidity. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: Prepare a solution with a concentration of 18.6 mg/mL of edetate disodium in water and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has
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dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken by the formula: W/ArV = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer a measured amount of Oral Suspension, previously well-shaken in its original container, equivalent to 800 mg of aluminum hydroxide, to a suitable beaker. Add 20 mL of water, stir, and slowly add 10 mL of hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 10 mL of Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: NLT 90.0% and NMT 115.0% MAGNESIUM HYDROXIDE Sample solution: Transfer an amount of Oral Suspension, previously well-shaken in its original container, equivalent to 800 mg of aluminum hydroxide, to a suitable beaker. Add 20 mL of water, stir, and slowly add 10 mL of hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet a volume of the Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker, add 200 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol, and mixing). Cool the solution to between 3 and 4 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting water for the Sample solution, and make any necessary correction. Each mL of 0.05 M edetate disodium consumed is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: NLT 90.0% and NMT 115.0% POLYDIMETHYLSILOXANE Standard solution: Prepare similarly to the Sample solution, except dissolve 50 mg of USP Polydimethylsiloxane RS in 25.0 mL of toluene, add 40 mL of 0.1 N sodium hydroxide, and add a volume of water equal to that of the specimen of Oral Suspension taken for the Sample solution. Sample solution: Transfer an amount of Oral Suspension equivalent to 50 mg of simethicone to a suitable round, narrow-mouth, screw-capped, 120-mL bottle, add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 25.0 mL W

of toluene, close the bottle securely with a cap having an inert liner, and shake for 15 min on a reciprocating shaker (e.g., about 200 oscillations per minute and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator. Remove about 5 mL of the upper, organic layer to a screw-capped, 15-mL test tube containing 0.5 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. The clear supernatant is the Sample solution. Blank: Mix 10 mL of toluene with 0.5 g of anhydrous sodium sulfate and centrifuge to obtain a clear supernatant. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: IR spectrophotometry Analytical wavelength: 7.9 m Cell: 0.5 mm Analysis Samples: Standard solution, Blank, and Sample solution Calculate the amount in mg of [(CH3)2 SiO]n in each mL of the Oral Suspension taken: Result = (AU/AS) (W/V) 100 = absorbance of the Sample solution = absorbance of the Standard solution = weight of USP Polydimethylsiloxane RS used in the Standard solution (mg) V = volume of Oral suspension used in the Sample solution (mL) Acceptance criteria: NLT 85.0% and NMT 115.0% SPECIFIC TESTS Microbial Enumeration Tests 61 and Tests for Specified Microorganisms 62 Acceptance criteria: The total aerobic microbial count is NMT 100 cfu/g and it meets the requirements of the test for the absence of Escherichia coli. ACID-NEUTRALIZING CAPACITY 301 Acceptance criteria: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: 0.55(0.0385A) + 0.8(0.0343M) 0.0385 = theoretical acid-neutralizing capacity of Al(OH)3 (mEq) A = quantity of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) 0.0343 = theoretical acid-neutralizing capacity of Mg(OH)2 (mEq) M = quantity of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg) PH 791 Acceptance criteria: 7.08.6 SODIUM CONTENT Potassium chloride solution: 38 mg/mL of potassium chloride Sodium chloride stock solution: 25.42 g/mL of sodium chloride (previously dried at 105 for 2 h) in water [NOTEThe solution contains 10 g/mL of sodium.] Standard solutions: On the day of use, transfer 4.0 mL of 1 N hydrochloric acid and 10.0 mL of Potassium chloride solution to each of two 100-mL volumetric flasks. To the respective flasks, add 5.0 mL and 10.0 mL of Sodium chloride stock solution. Dilute with water to volume. The resulting Standard solutions contain 0.5 and 1.0 g/mL of sodium (Na), respectively. Sample solution: Transfer 5.0 mL of Oral Suspension, previously well-shaken in its original container, to a 100-mL volumetric flask, add 50 mL of 1 N hydrochloric acid, boil for 15 mins, cool to room temperature, dilute with water to AU AS W
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C. PROCEDURE Sample: Wash the precipitate obtained in Identification test B with hot ammonium chloride solution (1 in 50). Analysis: Dissolve the precipitate in hydrochloric acid. Divide this solution into two portions. Acceptance criteria: The dropwise addition of 6 N ammonium hydroxide to one portion yields a gelatinous white precipitate, which does not dissolve in an excess of 6 N ammonium hydroxide. The dropwise addition of 1 N sodium hydroxide to the other portion yields a gelatinous white precipitate, which dissolves in an excess of 1 N sodium hydroxide, leaving some turbidity. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer the equivalent to 800 mg of aluminum hydroxide from Chewable Tablets (finely powder NLT 20 Chewable Tablets), to a 150-mL beaker. Add 20 mL of water, stir, and slowly add 30 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool to room temperature, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker. Add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from greenviolet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and making any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%115.0% MAGNESIUM HYDROXIDE Sample solution: Prepare as directed in the Assay for Aluminum Hydroxide. Analysis: Pipet a volume of the Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker, add 200 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of W
volume and filter, discarding the first few mL of the filtrate. Transfer 5.0 mL of the filtrate to a 100-mL volumetric flask containing 10.0 mL of Potassium chloride solution, and dilute with water to volume. Blank solution: Combine 4.0 mL of 1 N hydrochloric acid and 10.0 mL of Potassium chloride solution in a 100-mL volumetric flask, and dilute with water to volume. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption [NOTEUse an airacetylene flame.] Analytical wavelength: 589.0 nm Lamp: Sodium hollow-cathode lamp Analysis: Standard solutions, Blank solution, and Sample solution Plot the absorbances of the Standard solutions versus concentration, in g/mL, of sodium, and draw the straight line between the plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of sodium in the Sample solution. Calculate the quantity, in mg, of sodium in each mL of Oral Suspension taken: 0.4 C C = concentration of sodium in the Sample solution (g/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing. LABELING: Oral Suspension may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3. Label it to state the sodium content if it is greater than 1 mg per mL. USP REFERENCE STANDARDS 11 USP Polydimethylsiloxane RS
Alumina, Magnesia, and Simethicone Tablets

(Comment on this Monograph)id=m1753=Alumina, Magnesia, and Simethicone Tablets=A-Monos.pdf) (Current titlenot to change until February 1, 2010) Monograph title changeto become official February 1, 2010 See Alumina, Magnesia, and Simethicone Chewable Tablets DEFINITION Alumina, Magnesia, and Simethicone Chewable Tablets contain the equivalent of NLT 90.0% and NMT 115.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane [(CH3)2 SiO]n that is NLT 85.0% and NMT 115.0% of the labeled amount of simethicone. IDENTIFICATION A. INFRARED ABSORPTION 197S Cell: 0.5 mm Solution: Prepared as directed in the Assay for Polydimethylsiloxane. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Equivalent to 600 mg of magnesium hydroxide from finely powdered Chewable Tablets Add 25 mL of 3 N hydrochloric acid and 25 mL of water, and mix. Boil gently for 2 min. Allow to cool, and filter. Add 5 drops of methyl red TS, heat to boiling, and add 6 N ammonium hydroxide until the color of the solution just turns to deep yellow. Continue boiling for 2 min, and filter: the filtrate so obtained meets the requirements.
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dehydrated alcohol, and mixing). Cool the solution to 3 to 4 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting water for the Sample solution, and make any necessary correction. Each mL of 0.05 M edetate disodium consumed is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%115.0% POLYDIMETHYLSILOXANE Sample solution: Transfer the equivalent to 33 mg of simethicone from Chewable Tablets (finely powder NLT 20 Chewable Tablets), to a suitable round, narrow-mouth, screw-capped, 120-mL bottle. Add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 20.0 mL of toluene, close the bottle securely with a cap having an inert liner, and shake for 30 min on a reciprocating shaker (e.g., 200 oscillations per minute and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator, and allow to separate. Remove the upper, organic layer to a screw-capped, centrifuge tube containing 2 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. Standard solution: Similarly prepare a Standard solution, using 33 mg of USP Polydimethylsiloxane RS. Blank: Mix 10 mL of toluene with 0.5 g of anhydrous sodium sulfate, and centrifuge to obtain a clear supernatant. Analysis Samples: Sample solution, Standard solution, and Blank Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: IR spectrophotometer Analytical wavelength: Wavelength of maximum absorbance at 7.9 m (1265.8 cm1) Cell: 0.5 mm Analysis: Use Blank to set the instrument. Calculate the quantity in mg of [(CH3)2 SiO]n in the portion of Chewable Tablets taken: Result = (AU/AS) W = absorbance of the Sample solution = absorbance of the Standard solution = weight in mg of USP Polydimethylsiloxade RS used to prepare the Standard solution Acceptance criteria: 85.0%115.0% of simethicone AU AS W PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to aluminum hydroxide and to magnesium hydroxide SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Analysis: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq A = quantity of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2, 0.0343 mEq M = quantity of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg) DEFOAMING ACTIVITY Foaming solution: 500 g of FD&C Blue No. 1 and 1 g of octoxynol 9 in 100 mL of 0.1 N hydrochloric acid ANC1

Analysis: [NOTEFor each test, use a clean, unused, 250-mL glass jar.] Transfer a quantity of finely powdered Chewable Tablets, passed completely through an 80-mesh sieve, equivalent to 20 mg of simethicone, to a clean, unused, cylindrical 250-mL glass jar, fitted with a 50-mm cap, containing 100 mL of Foaming solution that has been warmed to 37. Cap the jar, and clamp it in an upright position on a wrist-action shaker. Using a radius of 13.3 0.4 cm (measured from center of shaft to center of bottle), shake for 10 s through an arc of 10 at a frequency of 300 30 strokes/min. Record the time required for the foam to collapse. The time, in seconds, for foam collapse is determined at the instant the first portion of foam-free liquid surface appears, measured from the end of the shaking period. Acceptance criteria: The defoaming activity time does not exceed 45 s. SODIUM CONTENT Potassium chloride solution: 38 mg/mL of potassium chloride Sodium chloride stock solution: Dissolve sodium chloride, previously dried at 105 for 2 h and weighed, in water, and dilute with water to obtain a solution containing 25.42 g/mL (10 g of sodium/mL). Standard solutions: On the day of use, transfer 4.0 mL of 1 N hydrochloric acid and 10.0 mL of Potassium chloride solution to each of two 100-mL volumetric flasks. To the respective flasks add 5.0 mL and 10.0 mL of Sodium chloride stock solution. Dilute with water to volume. These solutions contain 0.5 g/mL and 1.0 g/mL of sodium, respectively. Sample solution: Transfer the equivalent to the average weight of 1 Tablet from Chewable Tablets (weigh and finely powder NLT 20 Chewable Tablets), to a 100-mL volumetric flask. Add 50 mL of 1 N hydrochloric acid, boil for 15 min, cool to room temperature, and dilute with water to volume. Filter, discarding the first few mL of the filtrate. Transfer 5.0 mL of the filtrate to a 100-mL volumetric flask containing 10.0 mL of Potassium chloride solution, and dilute with water to volume. Blank: 1 N hydrochloric acid, Potassium chloride solution, and water (4:10:86) Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Analytical wavelength: 589.0 nm, sodium emission line Lamp: Sodium hollow-cathode lamp Flame: Air acetylene Analysis Samples: Standard solutions and Sample solution Plot the absorbances of the Standard solutions versus concentration, in g/mL, of sodium, and draw the straight line between the plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of sodium in the Sample solution. Calculate the quantity, in mg, of sodium per Tablet taken: Result = 2C ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Chewable Tablets to indicate that they are to be chewed before being swallowed. Label the Chewable Tablets to state the sodium content if it is greater than 5 mg/Tablet. The Chewable Tablets may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
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Ar V
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= atomic weight of aluminum, 26.98 = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer the equivalent to 800 mg of aluminum hydroxide from Chewable Tablets (finely powder NLT 20 Chewable Tablets), to a 150-mL beaker. Add 20 mL of water, stir, and slowly add 30 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool to room temperature, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from greenviolet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and making any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%115.0% MAGNESIUM HYDROXIDE Sample solution: Prepare as directed in the Assay for Aluminum Hydroxide. Analysis: Pipet a volume of the Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker, add 200 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol and mixing). Cool the solution to 34 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting water for the Sample solution, and make any necessary correction. Each mL of 0.05 M edetate disodium consumed is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%115.0% POLYDIMETHYLSILOXANE Sample solution: Transfer the equivalent to 33 mg of simethicone from Chewable Tablets (finely powder NLT 20 Chewable Tablets), to a suitable round, narrow-mouth, screw-capped, 120-mL bottle. Add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 20.0 mL of toluene, close the bottle securely with a cap having an inert liner, and shake for 30 min on a reciprocating shaker (e.g., 200 oscillations/min and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator, and allow to separate. Remove the upper, organic layer to a screw-capped, centrifuge tube containing 2 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. Standard solution: Similarly prepare a Standard solution, using 33 mg of USP Polydimethylsiloxane RS. Blank: Mix 10 mL of toluene with 0.5 g of anhydrous sodium sulfate, and centrifuge to obtain a clear supernatant. Analysis Samples: Sample solution, Standard solution, and Blank Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: IR spectrophotometer Analytical wavelength: Wavelength of maximum absorbance at 7.9 m (1265.8 cm1)
USP REFERENCE STANDARDS 11 USP Polydimethylsiloxane RS
Alumina, Magnesia, and Simethicone Chewable Tablets

(Comment on this Monograph)id=m1753=Alumina, Magnesia, and Simethicone Chewable Tablets=A-Monos.pdf) (Monograph under this new titleto become official February 1, 2010) (Current monograph title is Alumina, Magnesia, and Simethicone Tablets.) DEFINITION Alumina, Magnesia, and Simethicone Chewable Tablets contain the equivalent of NLT 90.0% and NMT 115.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane [(CH3)2 SiO]n that is NLT 85.0% and NMT 115.0% of the labeled amount of simethicone. IDENTIFICATION A. INFRARED ABSORPTION 197S Cell: 0.5 mm Solution: Prepared as directed in the Assay for Polydimethylsiloxane. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Equivalent to 600 mg of magnesium hydroxide from finely powdered Chewable Tablets Add 25 mL of 3 N hydrochloric acid and 25 mL of water, and mix. Boil gently for 2 min. Allow to cool, and filter. Add 5 drops of methyl red TS, heat to boiling, and add 6 N ammonium hydroxide until the color of the solution just turns to deep yellow. Continue boiling for 2 min, and filter: the filtrate so obtained meets the requirements. C. PROCEDURE Sample: Wash the precipitate obtained in Identification test B with hot ammonium chloride solution (1 in 50). Analysis: Dissolve the precipitate in hydrochloric acid. Divide this solution into two portions. Acceptance criteria: The dropwise addition of 6 N ammonium hydroxide to one portion yields a gelatinous white precipitate, which does not dissolve in an excess of 6 N ammonium hydroxide. The dropwise addition of 1 N sodium hydroxide to the other portion yields a gelatinous white precipitate, which dissolves in an excess of 1 N sodium hydroxide, leaving some turbidity. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV W = weight of aluminum in the portion of solution taken (mg)
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Cell: 0.5 mm Analysis: Use the Blank to set the instrument. Calculate the quantity in mg, of [(CH3)2 SiO]n in the portion of Chewable Tablets taken: Result = (AU/AS) W = absorbance of the Sample solution = absorbance of the Standard solution = weight in mg of USP Polydimethylsiloxane RS in Standard solution Acceptance criteria: 85.0%115.0% of simethicone AU AS W PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to aluminum hydroxide and to magnesium hydroxide SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Analysis: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) = theoretical acid-neutralizing capacity of Al(OH)3, 0.0385 mEq A = quantitly of Al(OH)3 in the specimen tested, based on the labeled quantity (mg) ANC2 = theoretical acid-neutralizing capacity of Mg(OH)2, 0.0343 mEq M = quantity of Mg(OH)2 in the specimen tested, based on the labeled quantity (mg) DEFOAMING ACTIVITY Foaming solution: 500 g of FD&C Blue No. 1 and 1 g of octoxynol 9 in 100 mL of 0.1 N hydrochloric acid. Analysis: [NOTEFor each test, use a clean, unused, 250-mL glass jar.] Transfer a quantity of finely powdered Chewable Tablets, passed completely through an 80-mesh sieve, equivalent to 20 mg of simethicone, to a clean, unused, cylindrical 250-mL glass jar, fitted with a 50-mm cap, containing 100 mL of Foaming solution that has been warmed to 37. Cap the jar, and clamp it in an upright position on a wrist-action shaker. Using a radius of 13.3 0.4 cm (measured from center of shaft to center of bottle), shake for 10 s through an arc of 10 at a frequency of 300 30 strokes/min. Record the time required for the foam to collapse. The time, in seconds, for foam collapse is determined at the instant the first portion of foam-free liquid surface appears, measured from the end of the shaking period. Acceptance criteria: The defoaming activity time does not exceed 45 s. SODIUM CONTENT Potassium chloride solution: 38 mg/mL of potassium chloride Sodium chloride stock solution: Dissolve sodium chloride, previously dried at 105 for 2 h and weighed, in water, and dilute with water to obtain a solution containing 25.42 g/mL (10 g of sodium/mL). Standard solutions: On the day of use, transfer 4.0 mL of 1 N hydrochloric acid and 10.0 mL of Potassium chloride solution to each of two 100-mL volumetric flasks. To the respective flasks, add 5.0 mL and 10.0 mL of Sodium chloride stock solution. Dilute with water to volume. These solutions contain 0.5 g/mL and 1.0 g/mL of sodium, respectively. Sample solution: Transfer the equivalent to the average weight of 1 Tablet from Chewable Tablets (weigh and finely powder NLT 20 Chewable Tablets), to a 100-mL volumetric flask ANC1

Add 50 mL of 1 N hydrochloric acid, boil for 15 min, cool to room temperature, and dilute with water to volume. Filter, discarding the first few mL of the filtrate. Transfer 5.0 mL of the filtrate to a 100-mL volumetric flask containing 10.0 mL of Potassium chloride solution, and dilute with water to volume. Blank: 1 N hydrochloric acid, Potassium chloride solution, and water (4:10:86) Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Analytical wavelength: 589.0 nm, sodium emission line Lamp: Sodium hollow-cathode lamp Flame: Air acetylene Analysis Samples: Standard solutions and Sample solution Plot the absorbances of the Standard solutions versus concentration, in g/mL, of sodium, and draw the straight line between the plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of sodium in the Sample solution. Calculate the quantity, in mg, of sodium per Tablet taken: Result = 2C ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Chewable Tablets to indicate that they are to be chewed before being swallowed. Label the Chewable Tablets to state the sodium content if it is greater than 5 mg/Tablet. The Chewable Tablets may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3. USP REFERENCE STANDARDS 11 USP Polydimethylsiloxane RS
Alumina and Magnesium Carbonate Oral Suspension

(Comment on this Monograph)id=m1770=Alumina and Magnesium Carbonate Oral Suspension=A-Monos.pdf) DEFINITION Alumina and Magnesium Carbonate Oral Suspension contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium carbonate (MgCO3). IDENTIFICATION A. PROCEDURE: Place 1 g in a flask equipped with a stopper and glass tubing, the tip of which is immersed in calcium hydroxide TS in a test tube. Add 5 mL of 3 N hydrochloric acid to the flask, and immediately insert the stopper: gas evolves in the flask and a precipitate is formed in the test tube. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: To a solution of 5 g in 10 mL of 3 N hydrochloric acid, add 5 drops of methyl red TS, heat to boiling, add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, then continue boiling for 2 min, and filter: the filtrate meets the requirements. C. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the precipitate obtained in Identification test B with a hot solution of ammonium chloride (1 in 50), and dissolve the precipitate in hydrochloric acid: the solution meets the requirements.
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Lamp: Magnesium hollowcathode Flame: Airacetylene Blank: Water Analysis Samples: Standard solutions and Sample solution Calculate the percentage of Mg(OH)2 in the portion of Oral Suspension taken: Result = (AU/AS) (CS/CU) (Mr/Ar) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution nominal concentration, in g of magnesium carbonate per mL, of the Sample solution, based on the labeled amount of magnesium carbonate in the portion of Oral Suspension taken (mg) and the extent of dilution = molecular weight of magnesium carbonate, Mr 84.31 = atomic weight of magnesium, 24.31 Ar Acceptance criteria: 90.0%110.0% AU AS CS CU SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: Its total aerobic microbial count does not exceed 100 cfu/mL, and it meets the requirements of the test for absence of Escherichia coli, Salmonella species, Staphylococcus aureus, and Pseudomonas aeruginosa. PH 791: 7.59.5 ACID-NEUTRALIZING CAPACITY 301 Analysis: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and NLT the number of mEq calculated: Result = 0.55(ANC1 A) + 0.8(ANC2 M) ANC1 A ANC2 M = theoretical acid-neutralizing capacity of Al(OH)3 (mEq), 0.0385 = quantity of Al(OH)3 in the specimen tested (mg), based on the labeled quantity = theoretical acid-neutralizing capacity of MgCO3 (mEq), 0.024 = quantity of MgCO3 in the specimen tested (mg), based on the labeled quantity = = = =
ASSAY ALUMINUM HYDROXIDE Potassium chloride solution: 45 mg/mL of potassium chloride solution Aluminum stock solution: Transfer 1.000 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of 6 N hydrochloric acid. Swirl to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Standard solutions: To separate 100-mL volumetric flasks, each containing 10 mL of Potassium chloride solution, transfer 9.0, 10.0, and 11.0 mL, respectively, of Aluminum stock solution, and dilute with water to volume. These Standard solutions contain 90.0, 100.0, and 110.0 g of aluminum per mL, respectively. Sample solution: Transfer a quantity of Oral Suspension, previously shaken in its original container, equivalent to 75 mg of aluminum hydroxide, to a suitable beaker. Add 25 mL of 6 N hydrochloric acid, and heat on a steam bath for 30 min, with occasional swirling. Cool, and transfer with the aid of water to a 250-mL volumetric flask containing 25 mL of Potassium chloride solution. Dilute with water to volume and filter. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Analytical wavelength: Aluminum emission line at 309.3 nm Lamp: Aluminum hollowcathode Flame: Nitrous oxideacetylene Blank: Water Analysis Samples: Standard solutions and Sample solution Calculate the quantity in mg, of Al(OH)3 in the portion of Oral Suspension taken: Result = Mr/Ar 0.25 AU/RS = the molecular weight of aluminum hydroxide, 78.00 = the atomic weight of aluminum, 26.98 Ar AU = absorbances from the Sample solution = average of the ratio of the absorbances of RS Standard solution to their respective concentration of Al in g/mL Acceptance criteria: 90.0%110.0% MAGNESIUM CARBONATE Lanthanum chloride solution: 5 mg/mL of lanthanum chloride in water Magnesium stock solution: Transfer 1.000 g of magnesium metal to a 1000-mL volumetric flask containing 50 mL of water, and slowly add 10 mL of hydrochloric acid. Dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Standard solutions: To separate 100-mL volumetric flasks, each containing 10 mL of Lanthanum chloride solution, transfer 1.70 mL and 1.80 mL, respectively, of Magnesium stock solution, and dilute with water to volume. These Standard solutions contain 1.7 g/mL of magnesium and 1.8 g/mL of magnesium, respectively. Sample solution: Quantitatively dilute a measured volume of the Sample solution prepared as directed in the Assay for Aluminum hydroxide with water to obtain a solution having a concentration of 6 g/mL of magnesium carbonate. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Analytical wavelength: Magnesium emission line at 285.2 nm Mr
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing.
Alumina and Magnesium Carbonate Tablets

(Comment on this Monograph)id=m1780=Alumina and Magnesium Carbonate Tablets=A-Monos.pdf) DEFINITION Alumina and Magnesium Carbonate Tablets contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium carbonate (MgCO3). IDENTIFICATION A. PROCEDURE Analysis: Place 1 g of finely powdered Tablets in a flask equipped with a stopper and glass tubing, the tip of which is immersed in calcium hydroxide TS in a test tube. Add 5
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mL of 3 N hydrochloric acid to the flask, and immediately insert the stopper Acceptance criteria: Gas evolves in the flask and a precipitate is formed in the test tube. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: To a 7-g portion of finely powdered Tablets, add 10 mL of 3 N hydrochloric acid and 5 drops of methyl red TS, heat to boiling, and add 6 N ammonium hydroxide until the color of the solution changes to deep yellow. Continue boiling for 2 min, and filter: the filtrate meets the requirements. C. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the precipitate obtained in Identification test B with a hot solution of ammonium chloride (1 in 50), and dissolve the precipitate in hydrochloric acid: the solution meets the requirements. ASSAY ALUMINUM HYDROXIDE Potassium chloride solution: 38.1 mg/mL of potassium chloride solution Digestion fluid: Hydrochloric acid, nitric acid, and water (1:2:2) [NOTEUse promptly] Aluminum stock solution: Transfer 1.000 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of 6 N hydrochloric acid. Swirl to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Standard solutions: To separate 100-mL volumetric flasks transfer 3.0, 4.0, and 5.0 mL of Aluminum stock solution, respectively. To each flask add 10 mL of Potassium chloride solution and 7.5 mL of Digestion fluid, dilute with water to volume, and mix. These Standard solutions contain 30, 40, and 50 g/mL of aluminum, respectively. Sample solution: Transfer an equivalent to 30 mg of aluminum hydroxide, from finely powdered Tablets (NLT 20), to a 100-mL volumetric flask, add 25-mL of Digestion fluid, and heat on a steam bath for 30 min or on a hot plate until the volume is reduced by about one-half. Cool, and dilute with water to volume. Filter, discarding the first 20 mL of the filtrate. Transfer 15.0 mL of the filtrate to a 50-mL volumetric flask, add 5.0 mL of Potassium chloride solution, and dilute with water to volume. [NOTEReserve a portion of the filtrate for use in the Assay for Magnesium carbonate.] Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Analytical wavelength: Aluminum emission line at 309.3 nm Lamp: Aluminum hollow-cathode Flame: Nitrous oxideacetylene Blank: Water Analysis Samples: Standard solutions and Sample solution Analysis: Plot the absorbances of the Standard preparations versus concentration, in g/mL, of aluminum, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of aluminum in each mL of the Sample solution. Calculate the percentage of Al(OH)3 in the portion of Tablets taken: Result = (C/3) (Mr/Ar) (100/L) C Mr Ar = concentration for the Sample solution (g/mL) = molecular weight of aluminum hydroxide, 78.00 = atomic weight of aluminum, 26.98

L = label claim (mg/unit) Acceptance criteria: 90.0%110.0% of the labeled amounts of Al(OH)3 MAGNESIUM CARBONATE Lanthanum chloride solution: Transfer 17.6 g of lanthanum chloride to a 1000-mL volumetric flask, add 500 mL of water, and carefully add 50 mL of hydrochloric acid. Mix, and allow to cool. Dilute with water to volume. Digestion fluid: Hydrochloric acid, nitric acid, and water (1:2:2) [NOTEUse promptly] Magnesium stock solution: Transfer 1.000 g of magnesium metal to a 1000-mL volumetric flask containing 50 mL of water, and slowly add 10 mL of hydrochloric acid. Dilute with water to volume. Transfer 5.0 mL of this solution to a 500-mL volumetric flask, and dilute with water to volume. Standard solutions: To separate 100-mL volumetric flasks, transfer 4.0, 6.0, and 8.0 mL of Magnesium stock solution, respectively. To each flask, add 0.5 mL of Digestion fluid and 10 mL of Lanthanum chloride solution, and dilute with water to volume. These Standard solutions contain 0.40, 0.60, and 0.80 g/mL of magnesium, respectively. Sample solution: Transfer a measured volume of the filtrate used to prepare the Sample solution in the Assay for Aluminum hydroxide, equivalent to 0.4 mg of magnesium carbonate, to a 200-mL volumetric flask, add 20 mL of Lanthanum chloride solution, and dilute with water to volume. Spectrometric conditions Mode: Atomic absorption spectrophotometer (see Spectrophotometry and Light-Scattering 851) Analytical wavelength: Magnesium emission line at 285.2 nm Lamp: Magnesium hollowcathode Flame: Airacetylene Blank: Water Analysis Samples: Standard solutions and Sample solution Analysis: Plot the absorbances of the Standard solutions versus concentration, in g/mL, of magnesium, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of magnesium in each mL of the Sample solution. Calculate the quantity in mg, of MgCO3 in the portion of Tablets taken: Result = (20C/V) (Mr/Ar) = concentration of the Sample solution (g/mL) = sample volume taken (mL) = molecular weight of magnesium carbonate, 84.31 = atomic weight of magnesium, 24.31 Ar Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701 Medium: Simulated gastric fluid TS being substituted for water in the test Time: 10 min UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to aluminum hydroxide and to magnesium carbonate SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling. C V Mr
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and slowly add 30 mL 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and dilute with water to volume. Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat near the boiling point for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM CARBONATE Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 750 mg of magnesium carbonate, to a 250-mL conical flask fitted with a two-hole stopper. Analysis: Fill the lower transverse section of a U-shaped drying tube of about 15-mm internal diameter and 15-cm height with loosely packed glass wool. Place in one arm of the tube, about 5 g of anhydrous calcium chloride, and weigh the tube and the contents. Into the other arm of the tube, place 9.5 g to 10.5 g of soda lime, and again weigh. Insert stoppers in the open arms of the U-tube, and connect the side tube of the arm filled with soda lime to a calcium chloride drying tube, which in turn is connected to one of the holes in the stopper of the 250-mL conical flask. Attach a dropping funnel to the other hole in the stopper of the 250-mL conical flask. Add 100 mL of water and 10 mL of a mixture of hydrochloric acid and nitric acid (4:1) to the 250mL conical flask through the dropping funnel, and close the dropping funnel. Heat the 250-mL conical flask at 95 for 1 h, and allow the evolved carbon dioxide to pass through the U-tube. Replace the dropping funnel with a source of carbon dioxide-free air, and pass the carbon dioxide-free air through the apparatus at a rate of about 75 mL/min for 30 min. Disconnect the U-tube, cool to room temperature, remove the stoppers, and weigh. The increase in weight corresponds to the quantity of carbon dioxide evolved. Calculate the quantity, in mg, of magnesium carbonate in each Tablet taken: Result = (Mr1/Mr2) (I) (WA/WP) = molecular weight of magnesium carbonate, 84.31 = molecular weight of carbon dioxide, 44.01 Mr2 I = quantity of carbon dioxide evolved from the portion of Tablets taken (mg) = average weight of 1 Tablet (g) WA = weight of the portion of Tablets taken (g) WP Acceptance criteria: 90.0%110.0% MAGNESIUM OXIDE Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 1000 mg of magnesium carbonate and magnesium oxide combined, to a beaker, add 20 mL of water, and slowly add 40 mL of 3 N hydrochloric acid, with mixing. Analysis: Heat the mixture to boiling, cool, and filter into a 200-mL volumetric flask. Wash the beaker with water, adding the washings to the filter. Add water to volume. Transfer 20.0 mL of this solution to a 400-mL beaker, add 180 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol Mr1
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Alumina, Magnesium Carbonate, and Magnesium Oxide Tablets

(Comment on this Monograph)id=m1795=Alumina, Magnesium Carbonate, and Magnesium Oxide Tablets=A-Monos.pdf) DEFINITION Alumina, Magnesium Carbonate, and Magnesium Oxide Tablets contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3] and magnesium carbonate (MgCO3), and NLT 85.0% and NMT 115.0% of the labeled amount of magnesium oxide (MgO). IDENTIFICATION A. PROCEDURE Place 3 g of finely powdered Tablets in a flask equipped with a stopper and glass tubing, the tip of which is immersed in calcium hydroxide TS in a test tube. Add 5 mL of 3 N hydrochloric acid to the flask, and immediately insert the stopper. Acceptance criteria: Gas evolves in the flask and a precipitate is formed in the test tube. B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Solution in the flask obtained in Identification test A Analysis: To the Sample solution add 5 drops of methyl red TS, and heat to boiling. Add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, continue boiling for 2 min, and filter through hardened filter paper. (Retain the filtrate for Identification test C.) Wash the precipitate with 350 mL of a hot ammonium chloride solution (1 in 50), discarding the washings. Acceptance criteria: The precipitate so obtained, dissolved in 3 N hydrochloric acid, meets the requirements of the tests for Aluminum. C. IDENTIFICATION TESTSGENERAL, Magnesium 191: The filtrate obtained in Identification test B meets the requirements of the tests for Magnesium. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 1200 mg of aluminum hydroxide, to a 150-mL beaker, add 20 mL of water, stir, W
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and mixing). Cool the solution to between 3 and 4 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting 20 mL of water for the assay solution, and make any necessary correction. Each mL of 0.05 M edetate disodium consumed is equivalent to 1.216 mg of Mg. Calculate the quantity, in mg, of magnesium equivalent in each Tablet taken: Result = 10T (WA/WP) T = magnesium equivalent obtained in the titration = average weight of 1 Tablet (g) WA = weight of the portion of Tablets taken (g) WP Calculate the quantity, in mg, of magnesium oxide in each Tablet taken: Result = (Mr3/Ar1) (A 0.2883B) = molecular weight of magnesium oxide, 40.30 = atomic weight of magnesium, 24.31 = quantity of magnesium equivalent in each Tablet (mg) B = quantity of magnesium carbonate in each Tablet, as determined in the Assay for magnesium carbonate (mg) Acceptance criteria: 85.0%115.0% Mr3 Ar1 A SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling. PERFORMANCE TESTS DISINTEGRATION 701: Medium: Simulated gastric fluid TS being substituted for water in the test Time: 10 min UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to alumina, to magnesium carbonate, and to magnesium oxide ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.

Acceptance criteria: The filtrate meets the requirements. C. PROCEDURE Transfer the filter paper and contents from Identification test B to a small platinum dish, ignite, cool in a desiccator, and weigh. Moisten the residue with water and add 6 mL of hydrofluoric acid. Evaporate to dryness, ignite for 5 min, cool in a desiccator, and weigh: a loss of more than 10% in relation to the weight of the residue from the initial ignition indicates SiO2. ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: W/ArV = weight of aluminum in the portion of solution taken (mg) V = volume of Edetate disodium titrant consumed (mL) = atomic weight of aluminum, 26.98 Ar Sample solution: Transfer 10 g of well-shaken Oral Suspension to a tared beaker, and weigh. Add 50 mL of water and 10 mL of hydrochloric acid, and digest on a steam bath for 1 h. Cool, and filter into a 200-mL volumetric flask, washing the filter with water into the flask. Dilute with water to volume. Analysis: Pipet 20 mL of Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from greenviolet to rose-pink. Perform a blank determination, substituting 20 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: Equivalent of 90.0%110.0% of the labeled amounts of Al(OH)3 MAGNESIUM TRISILICATE Sample solution: Prepare as directed in the Assay for Aluminum hydroxide. Analysis: Pipet 20 mL of Sample solution into a 400-mL beaker, add 180 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol). Cool the solution to between 3 and 4 by immersion of the beaker in an ice bath, then remove and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting 20 mL of water for the Sample solution, and make any necessary correction. W
Alumina and Magnesium Trisilicate Oral Suspension

(Comment on this Monograph)id=m1810=Alumina and Magnesium Trisilicate Oral Suspension=A-Monos.pdf) DEFINITION Alumina and Magnesium Trisilicate Oral Suspension contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum hydroxide [Al(OH)3], and magnesium trisilicate (Mg2Si3O8). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution To a mixture of 5 mL in 10 mL of 3 N hydrochloric acid add 5 drops of methyl red TS, heat to boiling, add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, then continue boiling for 2 min, and filter. Acceptance criteria: The filtrate meets the requirements. B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the solids on the filter obtained in Identification A with hot ammonium chloride solution (1 in 50), add 10 mL of 3 N hydrochloric acid, and filter.
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= weight of aluminum in the portion of solution taken = atomic weight of aluminum, 26.98 Ar V = volume of the Edetate disodium titrant consumed Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 600 mg of aluminum hydroxide, to a beaker, add 20 mL of water, stir, and slowly add 40 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and transfer to a 200-mL volumetric flask. Wash the beaker with water, adding the washings to the flask, and add water to volume. Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of 0.05 M Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, add 50 mL of alcohol and 2 mL of dithizone TS, and mix. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM TRISILICATE Potassium chloride solution: 50 mg/mL of potassium chloride in water Magnesium standard solution: Transfer 1.000 g of magnesium metal to a 1000-mL volumetric flask containing 50 mL of water, and slowly add 10 mL of hydrochloric acid. Dilute with water to volume. Transfer 5.0 mL of this solution to a 500-mL volumetric flask, and dilute with water to volume. Standard solutions: Transfer 16.0, 18.0, and 20.0 mL of Magnesium standard solution to separate 100-mL volumetric flasks, add 2.0 mL of Potassium chloride solution to each flask, and dilute with water to volume. These Standard solutions contain 1.6, 1.8, and 2.0 g/mL of magnesium respectively. [NOTEPrepare these solutions on the day of use.] Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 5 mg of magnesium trisilicate, to a 100-mL volumetric flask, and add 10 mL of 18 N sulfuric acid. Heat on a steam bath for 30 min with occasional swirling. Allow to cool, and dilute with water to volume. Filter this solution, discarding the first 20 mL of the filtrate. Transfer 20.0 mL of the filtrate to a second 100-mL volumetric flask, add 2.0 mL of Potassium chloride solution, and dilute with water to volume. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Analytical wavelength: 285.2 nm Lamp: Magnesium hollowcathode lamp Flame: Nitrous oxideacetylene flame Blank: Water Analysis Samples: Standard solutions and Sample solution Analysis: Plot the absorbances of the Standard solutions of magnesium, and draw the line best fitting the three plotted points in g/mL. From the graph so obtained, determine the concentration of magnesium in the Sample solution in g/mL. Calculate the quantity, in mg, of Mg2Si3O8 in the portion of Tablets taken: Result = 0.5 CU Mr/Ar CU Mr = concentration of the Sample solution (g/mL) = molecular weight of anhydrous magnesium trisilicate, 260.86
Each mL of 0.05 M edetate disodium consumed is equivalent to 6.521 mg of Mg2Si3O8. Acceptance criteria: Equivalent of 90.0%110.0% of the labeled amounts of Mg2Si3O8 SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling. PH 791: 7.58.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Alumina and Magnesium Trisilicate Tablets

(Comment on this Monograph)id=m1820=Alumina and Magnesium Trisilicate Tablets=A-Monos.pdf) DEFINITION Alumina and Magnesium Trisilicate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of [Al(OH)3] and Mg2Si3O8. IDENTIFICATION One powdered Tablet responds to the following tests. A. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: To a mixture of 5 mL in 10 mL of 3 N hydrochloric acid, add 5 drops of methyl red TS, heat to boiling, add 6 N ammonium hydroxide until the color of the solution changes to deep yellow, continue boiling for 2 min, and filter. Acceptance criteria: The filtrate meets the requirements. B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the solids on the filter obtained in Identification Test A with hot ammonium chloride solution (1 in 50), add 10 mL of 3 N hydrochloric acid, and filter. Acceptance criteria: The filtrate meets the requirements. C. PROCEDURE Sample: Transfer the filter paper and contents from Identification Test B to a small platinum dish, ignite, cool in a desiccator, and weigh. Analysis: Moisten the residue with water, and add 6 mL of hydrofluoric acid. Evaporate to dryness, ignite for 5 min, cool in a desiccator, and weigh: a loss of more than 10% in relation to the weight of the residue from the initial ignition indicates SiO2. Acceptance criteria: Meets the requirements ASSAY ALUMINUM HYDROXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution. Calculate the molarity of the portion of solution taken: Result = W/ArV
USP 32
Ar = twice the atomic weight of magnesium, 48.62 Acceptance criteria: 90.0%110.0% SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling. [NOTETablets labeled for the temporary relief of heartburn (acid indigestion) due to acid reflux are exempt from this requirement.] PH 791: NLT 4.5, determined on the foam layer obtained in the Foam test, where Tablets are labeled for the temporary relief of heartburn (acid indigestion) due to acid reflux. [NOTETake care that the electrodes do not touch the liquid beneath the foam.] FOAM [Where Tablets are labeled for the temporary relief of heartburn (acid indigestion) due to acid reflux] Sample solution: Finely powder a number of Tablets equivalent to the minimum single dose recommended in the labeling, and transfer the powder to a 100-mL beaker having an inside diameter of 45 mm. Add 5 mL of alcohol and sufficient water to make 40 mL. Analysis: Mix at 300 rpm for 60 s, using a magnetic stirrer and a 9.5- 38-mm polytef-coated stirring bar. Stop the stirrer, and carefully add 10 mL of 0.5 N hydrochloric acid down the side of the beaker. Stir for 30 s at 300 rpm. Allow to stand for 10 min, and measure the thickness of the foam layer above the liquid in the beaker. Acceptance criteria: The thickness of the foam is NLT 10 mm. PERFORMANCE TESTS DISINTEGRATION 701 Medium: Simulated gastric fluid TS being substituted for water in the test [NOTETablets that must be chewed before swallowing are exempt from this requirement.] Time: 10 min UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to aluminum hydroxide and to magnesium trisilicate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Tablets prepared with the use of Dried Aluminum Hydroxide Gel may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3. Tablets intended for the temporary relief of heartburn (acid indigestion) due to acid reflux are so labeled. Tablets that must be chewed before swallowing are so labeled.
Official Monographs / Aluminum 123

DEFINITION Aluminum Acetate Topical Solution yields, from each 100 mL, NLT 1.20 g and NMT 1.45 g of aluminum oxide (Al2O3), and NLT 4.24 g and NMT 5.12 g of acetic acid (C2H4O2), corresponding to NLT 4.8 g and NMT 5.8 g of aluminum acetate (C6H9AlO6). Aluminum Acetate Topical Solution may be stabilized by the addition of NMT 0.6% of Boric Acid.
Aluminum Subacetate Topical Solution Glacial Acetic Acid Purified Water To make 545 mL 15 mL A sufficient quantity 1000 mL
Add the Glacial Acetic Acid to the Aluminum Subacetate Topical Solution and sufficient water to make 1000 mL. Mix, and filter, if necessary. [NOTEDispense only clear Aluminum Acetate Topical Solution.] IDENTIFICATION IDENTIFICATION TESTSGENERAL, Aluminum AND Acetate 191 Acceptance criteria: Meets the requirements of the tests ASSAY ALUMINUM OXIDE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Standardization of Titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: W/ArV = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample: 25 mL of Aluminum Acetate Topical Solution Analysis: Pipet the Sample into a 250-mL volumetric flask, add 5 mL of hydrochloric acid, and dilute with water to volume. Pipet 25 mL of this solution into a 250-mL beaker, and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, then heat the solution near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting water for the Sample, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 2.549 mg of Al2O3. Acceptance criteria: NLT 1.20 g/100 mL and NMT 1.45 g/100 mL of aluminum oxide (Al2O3) ACETIC ACID Sample: 20 mL Aluminum Acetate Topical Solution Analysis: Pipet the Sample into a Kjeldahl flask containing a mixture of 20 mL of phosphoric acid and 150 mL of water. W
Aluminum Acetate Topical Solution

(Comment on this Monograph)id=m1890=Aluminum Acetate Topical Solution=A-Monos.pdf)
C6H9AlO6 Acetic acid, aluminum salt; Aluminum acetate [139-12-8].
204.11
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Aluminum / Official Monographs
USP 32
Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V W Ar V = weight of aluminum (mg) = atomic weight of aluminum, 26.98 = volume of Edetate disodium titrant consumed (mL) Sample solution: 20 mg/mL Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker, and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the sample, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 6.667 mg of AlCl3. Acceptance criteria: 95.0%102.0%
Connect the flask to a condenser, the delivery tube from which dips beneath the surface of 50.0 mL of 0.5 N sodium hydroxide VS contained in a receiving flask. Distill about 160 mL, then remove the delivery tube from below the surface of the liquid, allow the distilling flask to cool, add 50 mL of water, and distill an additional 40 to 45 mL into the receiving flask. Add phenolphthalein TS to the distillate, and titrate the excess 0.5 N sodium hydroxide VS with 0.5 N sulfuric acid VS. Each mL of 0.5 N sodium hydroxide is equivalent to 30.03 mg of C2H4O2. Acceptance criteria: NLT 4.24 g/100 mL and NMT 5.12 g/100 mL of C2H4O2 [NOTEThe results for both assays correspond to NLT 4.8 g/100 mL and NMT 5.8 g/100 mL of C6H9AlO6.] OTHER COMPONENTS LIMIT OF BORIC ACID Sample: 25 mL Aluminum Acetate Topical Solution Analysis: Pipet the Sample into 75 mL of water in a conical flask. Add 3 mL of phenolphthalein TS, then add 0.5 N sodium hydroxide VS from a buret until a faint pink color is obtained. Heat to boiling, and again neutralize. Add 150 mL of glycerin to the neutralized solution, and titrate with 0.5 N sodium hydroxide VS. Perform a blank determination in a similar manner. From the volume of 0.5 N sodium hydroxide VS used after the addition of the glycerin, subtract the volume used in the blank. Each mL of 0.5 N sodium hydroxide is equivalent to 30.92 mg of H3BO3. Acceptance criteria: NMT 0.6% IMPURITIES Inorganic Impurities HEAVY METALS 231 Sample solution: 2 mL of Sample diluted to 25 mL Acceptance criteria: NMT 10 ppm SPECIFIC TESTS PH 791 Acceptance criteria:
3.64.4
Aluminum Chloride
(Comment on this Monograph)id=m2040=Aluminum Chloride=A-Monos.pdf) AlCl3 6H2O Aluminum chloride, hexahydrate; Aluminum chloride hexahydrate [7784-13-6]. Anhydrous [7446-70-0]. 241.43 133.34
IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Sulfate 221 Sample solution: 10 mg/mL Analysis: Add 0.2 mL of barium chloride TS to 10 mL of the Sample solution. Acceptance criteria: No turbidity is produced within 1 min. IRON 241 Sample solution: Dissolve 1.0 g of sample in 45 mL of water, and add 2 mL of hydrochloric acid. Acceptance criteria: NMT 10 ppm HEAVY METALS, Method I 231 Sample solution: Dissolve 1 g of sample in 1 mL of 1 N acetic acid, and sufficient water to make 25 mL. Acceptance criteria: NMT 20 ppm SPECIFIC TESTS WATER DETERMINATION, Method I 921 Acceptance criteria: Between 42.0% and 48.0% LIMIT OF ALKALIES AND ALKALINE EARTHS Sample solution: 66.7 mg/mL Analysis: To 100 mL of boiling Sample solution, add a few drops of methyl red TS, then add 6 N ammonium hydroxide until the color of the solution just changes to a distinct yellow. Add hot water to restore the volume to 150 mL, and filter while hot. Evaporate 75 mL of the filtrate to dryness, and ignite to a constant weight. Acceptance criteria: The weight of the residue does not exceed 2.5 mg (0.5%).
DEFINITION Aluminum Chloride contains NLT 95.0% and NMT 102.0% of AlCl3, calculated on the anhydrous basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements IDENTIFICATION TESTSGENERAL, Chloride 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements ASSAY PROCEDURE Edetate disodium titrant: in water
18.6 mg/mL of edetate disodium
USP 32

of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V 210.48 174.45 = weight of aluminum (mg) = atomic weight of aluminum, 26.98 = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 200 mg of Aluminum Chlorohydrate to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). Use the aluminum content obtained to calculate the Aluminum/Chloride Atomic Ratio. ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in the test for Content of Aluminum by the percentage of chloride found in the test for Content of Chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 1.91:1 and 2.10:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Acceptance criteria: NMT 20 ppm LIMIT OF IRON Standard solution: 2.0 mL of Standard Iron Solution, prepared as directed for Iron 241 Sample solution: 27 mg/mL Analysis: Pipet 5.0 mL of the Standard solution into a 50mL beaker. Pipet 5.0 mL of the Sample solution to a second 50-mL beaker. To each of the beakers, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed for Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). W Ar V
Aluminum Chlorohydrate
(Comment on this Monograph)id=m2050=Aluminum Chlorohydrate=A-Monos.pdf) AIy(OH)3y-zClz H2O Aluminum chlorohydroxide; Aluminum hydroxychloride; Dihydrate [12042-91-0]. Anhydrous [1327-41-9]. Aluminum chlorohydroxide, dihydrate; Aluminum hydroxychloride, dihydrate; Dihydrate [12042-91-0]. Anhydrous [1327-41-9].
DEFINITION Aluminum Chlorohydrate consists of complex basic aluminum chloride that is polymeric and loosely hydrated and encompasses a range of aluminum-to-chloride atomic ratios between 1.91:1 and 2.10:1. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum chlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Chloride 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements ASSAY PROCEDURE Calculate the percentage of anhydrous aluminum chlorohydrate in the Aluminum Chlorohydrate taken: Result = Al({Ar1x + [Mr(3x -1)] + Ar2}/Ar1x) = percentage of aluminum as obtained in the test for Content of Aluminum Ar1 = atomic weight of aluminum, 26.98 x = Aluminum/Chloride Atomic Ratio Mr = molecular weight of the hydroxide anion (OH), 17.01 Ar2 = atomic weight of chlorine (Cl), 35.453 Acceptance criteria: NLT 90.0% and NMT 110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 700 mg of Aluminum Chlorohydrate Analysis: Transfer the Sample to a 250-mL beaker, and add, with stirring, 100 mL of water and 10 mL of diluted nitric acid. Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). Use the chloride content obtained to calculate the Aluminum /Chloride Atomic Ratio. CONTENT OF ALUMINUM Edetate disodium titrant: 37.2 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture Al
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ASSAY [NOTEConduct Procedure 1, Procedure 2, and Procedure 3.] PROCEDURE 1: CONTENT OF CHLORIDE Sample: 1.4 g of Aluminum Chlorohydrate Solution Analysis: Transfer the Sample to a 250-mL beaker, and add 100 mL of water and 10 mL of diluted nitric acid with stirring. Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). Calculate the percentage of chloride (Cl) found and use the chloride content thus obtained to calculate the Aluminum/ Chloride Atomic Ratio in Procedure 3. PROCEDURE 2: CONTENT OF ALUMINUM Edetate disodium titrant: 37.2 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS. Boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 400 mg of Aluminum Chlorohydrate Solution to a 250-mL beaker. Add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). Calculate the percentage of aluminum (Al) found and use the aluminum content thus obtained to calculate the Aluminum/Chloride Atomic Ratio (below). PROCEDURE 3: ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in Procedure 2: Content of Aluminum by the percentage of chloride found in Procedure 1: Content of Chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 1.91:1 and 2.10:1. W
SPECIFIC TESTS PH 791 Sample solution: 15 in 100 (w/w) Acceptance criteria: Between 3.0 and 5.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum chlorohydrate.
Aluminum Chlorohydrate Solution

(Comment on this Monograph)id=m2052=Aluminum Chlorohydrate Solution=A-Monos.pdf) DEFINITION Aluminum Chlorohydrate Solution consists of complex basic aluminum chloride that is polymeric and encompasses a range of aluminum-to-chloride ratios between 1.91:1 and 2.10:1. The following solvents may be used: water, propylene glycol, dipropylene glycol, or alcohol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum chlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191 Sample solution: Amount equivalent to 100 mg/mL of anhydrous aluminum chlorohydrate Acceptance criteria: Meets the requirements B. PROPYLENE GLYCOL [NOTEPerform this test where propylene glycol is stated on the label.] Sample solution: 2 g of Aluminum Chlorohydrate Solution in 10 mL of isopropyl alcohol [NOTEFilter the solution after preparation.] Analysis: Evaporate the Sample solution to about 1 mL on a steam bath. Acceptance criteria: The IR spectrum of a film of the resulting solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. DIPROPYLENE GLYCOL [NOTEPerform this test where dipropylene glycol is stated on the label.] Sample solution: 2 g of Aluminum Chlorohydrate Solution in 10 mL of isopropyl alcohol [NOTEFilter the solution after preparation.] Analysis: Evaporate the Sample solution to about 1 mL on a steam bath. Acceptance criteria: The IR spectrum of a film of the resulting solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. D. ALCOHOL [NOTEPerform this test where alcohol is stated on the label.] Analysis: In a small beaker, mix 5 drops of Aluminum Chlorohydrate Solution with 1 mL of potassium permanganate solution (1 in 100), and 5 drops of 2 N sulfuric acid. Immediately cover the beaker with filter paper moistened with a freshly prepared solution of 0.1 g of sodium nitroferricyanide and 0.25 g of piperazine in 5 mL of water. Acceptance criteria: An intense blue color is produced on the filter paper, the color fading after a few minutes.
USP 32
PROCEDURE 4 Analysis: Calculate the percentage of anhydrous aluminum chlorohydrate in the portion of Aluminum Chlorohydrate taken: Result = Al({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X) = percentage of aluminum as obtained in Procedure 2: Content of Aluminum = atomic weight of aluminum, 26.98 Ar1 X = aluminum/chloride atomic ratio as obtained in Procedure 3 = molecular weight of the hydroxide anion (OH), Mr 17.01 = atomic weight of chlorine (Cl), 35.453 Ar2 Acceptance criteria: NLT 90.0% and NMT 110.0% IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm HEAVY METALS, Method I 231: NMT 10 ppm LIMIT OF IRON Standard solution: 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241 Sample solution: Transfer 5.3 g of Aluminum Chlorohydrate Solution to a 100-mL volumetric flask, and dilute to volume with water. Analysis: Pipet 5.0 mL of the Standard solution into a 50-mL beaker. Pipet 5.0 mL of the Sample solution into a second 50-mL beaker. To each of the beakers, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (75 ppm). SPECIFIC TESTS PH 791 Sample solution: Dilute 3 g of the Aluminum Chlorohydrate Solution with water to 10 mL. Acceptance criteria: 3.05.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label Solution to state the solvent used and the claimed concentration of anhydrous aluminum chlorohydrate contained therein. Al%

IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements of the tests B. INFRARED ABSORPTION 197F Sample solution: Dissolve 0.5 g in 40 mL of water and, while mixing, adjust with 2.5 N sodium hydroxide to a pH of 9.55 0.05. Filter the suspension of precipitate thus obtained. Evaporate about 15 mL of the filtrate to about 1 mL on a hot plate. Deposit the solution on a silver chloride disk. Standard solution: A similar preparation of polyethylene glycol ASSAY PROCEDURE Analysis Calculate the percentage of anhydrous aluminum chlorohydrate in the Aluminum Chlorohydrex Polyethylene Glycol: Result = Al({AR1X + [MR(3X -1)] + AR2}/AR1X) = percentage of aluminum as obtained in the test for Content of Aluminum X = aluminum/chloride atomic ratio = atomic weight of aluminum, 26.98 AR1 = molecular weight of the hydroxide anion (OH), MR 17.01 = atomic weight of chlorine (Cl), 35.453 AR2 Acceptance criteria: NLT 90.0% and NMT 110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 700 mg of Aluminum Chlorohydrex Polyethylene Glycol Analysis: Transfer the Sample to a 250-mL beaker and add 100 mL of water and 10 mL of diluted nitric acid with stirring. Titrate with 0.1 N silver nitrate VS using a glass silver-silver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). Use the chloride content thus obtained to calculate the Aluminum/Chloride Atomic Ratio. CONTENT OF ALUMINUM Edetate disodium titrant: 37.2 mg/mL of edetate disodium Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V W Ar = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Al
Aluminum Chlorohydrex Polyethylene Glycol

(Comment on this Monograph)id=m2054=Aluminum Chlorohydrex Polyethylene Glycol=A-Monos.pdf) Aly(OH)3y-zClz nH2O mH(OCH2CH2)nOH Aluminum chlorohydroxide polyethylene glycol complex; Aluminum hydroxychloride polyethylene glycol complex. DEFINITION Aluminum Chlorohydrex Polyethylene Glycol consists of aluminum chlorohydrate in which some of the waters of hydration have been replaced by polyethylene glycol. It encompasses a range of aluminum-to-chloride atomic ratios between 1.91:1 and 2.10:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum chlorohydrate.
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DEFINITION Aluminum Chlorohydrex Propylene Glycol is a complex of aluminum chlorohydrate and propylene glycol in which some of the waters of hydration of the aluminum chlorohydrate have been replaced by propylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum chlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements B. INFRARED ABSORPTION 197F Sample solution: Dissolve 0.5 g in 40 mL of water and, while mixing, adjust with 2.5 N sodium hydroxide to a pH of 9.55 0.05. Filter the suspension of precipitate thus obtained. Evaporate about 15 mL of the filtrate to about 1 mL on a hot plate. Deposit the solution on a silver chloride disk. Standard solution: A similar preparation of propylene glycol ASSAY PROCEDURE Analysis Calculate the percentage of anhydrous aluminum chlorohydrate in the Aluminum Chlorohydrex Propylene Glycol: Result = Al({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X) = percentage of aluminum as obtained in the test for Procedure 2: Content of Aluminum (see below) = atomic weight of aluminum, 26.98 Ar1 X = Aluminum/Chloride Atomic Ratio = molecular weight of the hydroxide anion (OH), Mr 17.01 = atomic weight of chlorine (Cl), 35.453 Ar2 Acceptance criteria: NLT 90.0% and NMT 110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 700 mg of Aluminum Chlorohydrex Propylene Glycol Analysis: Transfer the Sample to a 250-mL beaker and add 100 mL of water and 10 mL of diluted nitric acid with stirring. Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). Use the chloride content thus obtained to calculate the Aluminum/Chloride Atomic Ratio. CONTENT OF ALUMINUM Edetate disodium titrant: 37.2 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, Al%
= volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 200 mg of Aluminum Chlorohydrex Polyethylene Glycol to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acid-ammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). Use the aluminum content thus obtained to calculate the Aluminum/Chloride Atomic Ratio. ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in the test for Content of Aluminum by the percentage of chloride found in the test for Content of Chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 1.91:1 and 2.10:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Acceptance criteria: NMT 20 ppm LIMIT OF IRON Standard solution: 2.0 mL of Standard Iron Solution Prepared as directed under Iron 241. Sample solution: 27 mg/mL Analysis: Pipet 5.0 mL of the Standard solution into a 50mL beaker. Pipet 5.0 mL of the Sample solution to a second 50-mL beaker. To each of the beakers, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). SPECIFIC TESTS PH 791 Sample solution: 15 in 100 (w/w) Acceptance criteria: Between 3.0 and 5.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum chlorohydrate.
Aluminum Chlorohydrex Propylene Glycol

(Comment on this Monograph)id=m2057=Aluminum Chlorohydrex Propylene Glycol=A-Monos.pdf) Aly(OH)3y-zClz nH2O mC3H8O2 Al2(H2O)y-z(OH)6-n(Cl)n(C3H8O2)z Aluminum chlorohydroxide, hydrate:propylene glycol complex (1:1); Aluminum hydroxychloride, hydrate:propylene glycol complex (1:1) [53026-85-0].
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substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight aluminum in the portion of solution taken (mg) Ar = atomic weight of aluminum (Al), 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 1.6 g of Aluminum Chlorohydrex Propylene Glycol to a 100-mL beaker, add 15 to 20 mL of water and 5 to 6 mL of hydrochloric acid, boil on a hot plate for 15 to 20 min, and allow to cool. With the aid of water, transfer to a 100-mL volumetric flask and dilute to volume. Analysis: Transfer 5.0 mL of the Sample solution to a 250mL beaker, add 10 to 15 mL of water and adjust with 1 N sodium hydroxide to a pH of 1.5 0.5. Add 10.0 mL of Edetate disodium titrant and heat to boiling. Cool the solution, and carefully introduce a magnetic stirring bar into the beaker. Add 10 to 15 mL of acetic acidammonium acetate buffer TS, 40 to 50 mL of alcohol, and, while stirring, adjust with glacial acetic acid to a pH of 4.6 0.1. Add 1 to 2 mL of dithizone TS and 40 to 50 mL of alcohol, and titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). Use the aluminum content thus obtained to calculate the Aluminum/Chloride Atomic Ratio. ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in the test for Content of Aluminum by the percentage of chloride found in the test for Content of Chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 1.91:1 and 2.1:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Acceptance criteria: NMT 20 ppm LIMIT OF IRON Standard solution: 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241 Sample solution: 27 mg/mL Analysis: Pipet 5.0 mL of the Standard solution into a 50mL beaker. Pipet 5.0 mL of the Sample solution to a second 50-mL beaker. To each of the beakers add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). SPECIFIC TESTS PH 791 Sample solution: 15 in 100 (w/w) Acceptance criteria: Between 3.0 and 5.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum chlorohydrate. W
Aluminum Dichlorohydrate
(Comment on this Monograph)id=m2060=Aluminum Dichlorohydrate=A-Monos.pdf) Aly(OH)3y-zClz nH2O Aluminum chlorohydroxide; Aluminum hydroxychloride. DEFINITION Aluminum Dichlorohydrate consists of complex basic aluminum chloride that is polymeric and loosely hydrated and encompasses a range of aluminum-to-chloride atomic ratios between 0.90:1 and 1.25:1. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum dichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Chloride 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements ASSAY PROCEDURE Analysis Calculate the percentage of anhydrous aluminum dichlorohydrate in the portion of Aluminum Dichlorohydrate taken: Result = Al({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X) = percentage of aluminum as obtained in the test for Content of Aluminum = atomic weight of aluminum, 26.98 Ar1 X = Aluminum/Chloride Atomic Ratio, as determined below = molecular weight of the hydroxide anion (OH), Mr 17.01 = atomic weight of chlorine (Cl), 35.453 Ar2 Acceptance criteria: NLT 90.0% and NMT 110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 700 mg of Aluminum Dichlorohydrate Analysis: Transfer the Sample to a 250-mL beaker and add 100 mL of water and 10 mL of diluted nitric acid with stirring. Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). Use the chloride content thus obtained to calculate the Aluminum/Chloride Atomic Ratio (below). CONTENT OF ALUMINUM Edetate disodium titrant: 37.2 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Al
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Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) Ar = atomic weight of aluminum, 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 200 mg of Aluminum Dichlorohydrate to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). Use the aluminum content thus obtained to calculate the Aluminum/Chloride Atomic Ratio (below). ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in Content of Aluminum (above) by the percentage of chloride found in Content of Chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 0.90:1 and 1.25:1. IMPURITIES ARSENIC, Method I 211 Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Acceptance criteria: NMT 20 ppm LIMIT OF IRON Standard solution: 2.0 mL of Standard Iron Solution, prepared as directed in Iron 241 Sample solution: 27 mg/mL Analysis: Pipet 5.0 mL of the Standard solution into a 50-mL beaker. Pipet 5.0 mL of the Sample solution to a second 50mL beaker. To each of the beakers, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution (prepared as directed in Iron 241), transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). SPECIFIC TESTS PH 791 Sample solution: 15 in 100 (w/w) Acceptance criteria: Between 3.0 and 5.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum dichlorohydrate. W
Aluminum Dichlorohydrate Solution

(Comment on this Monograph)id=m2062=Aluminum Dichlorohydrate Solution=A-Monos.pdf) DEFINITION Aluminum Dichlorohydrate Solution consists of complex basic aluminum chloride that is polymeric and encompasses a range of aluminum-to-chloride atomic ratios between 0.90:1 and 1.25:1. The following solvents may be used: water, propylene glycol, dipropylene glycol, or alcohol. It contains the equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum dichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum 191: Sample solution: Amount equivalent to 100 mg/mL of anhydrous aluminum dichlorohydrate Acceptance criteria: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Chloride 191: Sample solution: Amount equivalent to 100 mg/mL of anhydrous aluminum dichlorohydrate Acceptance criteria: Meets the requirements C. INFRARED ABSORPTION 197F [NOTEPerform this test where either propylene glycol or dipropylene glycol is stated on the label.] Standard solution: A similar preparation of propylene glycol or dipropylene glycol (where propylene glycol or dipropylene glycol are indicated on the label) Sample solution: Add 10 mL of isopropyl alcohol to 2 g of Aluminum Dichlorohydrate Solution, and filter. Evaporate the filtrate to about 1 mL on a steam bath. Deposit this solution on a silver chloride disk. D. IDENTIFICATION OF ALCOHOL [NOTEPerform this test where alcohol is stated on the label.] Analysis: Mix 5 drops of Aluminum Dichlorohydrate Solution in a small beaker with 1 mL of potassium permanganate solution (1 in 100) and 5 drops of 2 N sulfuric acid, and cover the beaker immediately with filter paper moistened with a freshly prepared solution of 0.1 g of sodium nitroferricyanide and 0.25 g of piperazine in 5 mL of water. Acceptance criteria: An intense blue color is produced on the filter paper, the color becoming paler after a few minutes. ASSAY PROCEDURE Calculate the percentage of anhydrous aluminum dichlorohydrate in the portion of Solution taken: Result = Al({Ar1x + [Mr(3x 1)] + Ar2}/Ar1x) Al Ar1 x Mr Ar2 = percentage of aluminum as obtained in the test for Content of Aluminum = atomic weight of aluminum, 26.98 = Aluminum/Chloride Atomic Ratio = molecular weight of the hydroxide anion (OH), 17.0 = atomic weight of chlorine (Cl), 35.453
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Acceptance criteria: NLT 90.0% and NMT 110.0%

Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Sample: Use a quantity of Aluminum Dichlorohydrate Solution Acceptance criteria: NMT 10 ppm LIMIT OF IRON Standard solution: 2.0 mL of Standard Iron Solution, prepared as directed for Iron 241 Sample solution: Transfer 5.3 g of Aluminum Dichlorohydrate Solution to a 100-mL volumetric flask, and dilute to volume with water. Analysis: Pipet 5.0 mL of the Standard solution into a 50mL beaker. Pipet 5.0 mL of the Sample solution into a second 50-mL beaker. To each of the beakers, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed for Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that from the Standard solution (75 ppm). SPECIFIC TESTS PH 791 Sample solution: Dilute 3 g of Aluminum Dichlorohydrate Solution with water to 10 mL. Acceptance criteria: Between 3.0 and 5.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label Aluminum Dichlorohydrate Solution to state the solvent used and the claimed concentration of anhydrous aluminum dichlorohydrate contained therein.
OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 1.4 g of Aluminum Dichlorohydrate Solution Analysis: Transfer the Sample to a 250-mL beaker, and add, with stirring, 100 mL of water and 10 mL of diluted nitric acid. Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). Calculate the percentage of chloride (Cl) found and use the chloride content thus obtained to calculate the Aluminum/ Chloride Atomic Ratio. CONTENT OF ALUMINUM Edetate disodium titrant: 37.2 mg/mL of edetate disodium in water Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum (mg) = atomic weight of aluminium, 26.98 = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 400 mg of Aluminum Dichlorohydrate Solution to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acid-ammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). Calculate the percentage of aluminum (Al) found and use the aluminum content obtained to calculate the Aluminum/ Chloride Atomic Ratio. ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in the test for Content of Aluminum by the percentage of chloride found in the test for Content of Chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 0.90:1 and 1.25:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Sample: Use a weighed quantity of Aluminum Dichlorohydrate Solution. W Ar V
Aluminum Dichlorohydrex Polyethylene Glycol

(Comment on this Monograph)id=m2064=Aluminum Dichlorohydrex Polyethylene Glycol=A-Monos.pdf) Aly(OH)3y-zClz nH2O mH(OCH2CH2)nOH. Aluminum chlorohydroxide polyethylene glycol complex; Aluminum hydroxychloride polyethylene glycol complex. DEFINITION Aluminum Dichlorohydrex Polyethylene Glycol consists of aluminum dichlorohydrate, in which some of the waters of hydration have been replaced by polyethylene glycol. It encompasses a range of aluminum-to-chloride atomic ratios between 0.90:1 and 1.25:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum dichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum 191: Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements of the test B. IDENTIFICATION TESTSGENERAL, Chloride 191: Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements of the test C. INFRARED ABSORPTION 197F Sample solution: Dissolve 0.5 g in 40 mL of water, and, while mixing, adjust with 2.5 N sodium hydroxide to a pH of 9.55 0.05. Filter the suspension of precipitate obtained. Evaporate about 15 mL of the filtrate to about 1 mL on a hot plate. Deposit the solution on a silver chloride disk. Standard solution: A similar preparation of polyethylene glycol
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ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in the test for Content of Aluminum by the percentage of chloride found in the test for Content of Chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: Between 0.90:1 and 1.25:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Acceptance criteria: NMT 20 ppm LIMIT OF IRON Standard solution: 2.0 mL of Standard Iron Solution, prepared as directed for Iron 241 Sample solution: 27 mg/mL Analysis: Pipet 5.0 mL of the Standard solution into a 50mL beaker. Pipet 5.0 mL of the Sample solution to a second 50-mL beaker. To each of the beakers, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed for Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that from the Standard solution (150 ppm). SPECIFIC TESTS PH 791 Sample solution: 15 in 100 (w/w) Acceptance criteria: Between 3.0 and 5.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum dichlorohydrate.
ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum dichlorohydrate in the Aluminum Dichlorohydrex Polyethylene Glycol taken: Result = Al({Ar1X + [Mr(3X1)] + Ar2}/Ar1X) = percentage of aluminum as obtained in the test for Content of Aluminum = atomic weight of aluminum, 26.98 Ar1 X = aluminum/chloride atomic ratio (as determined below) = molecular weight of hydroxide anion (OH), Mr 17.01 Ar2 = atomic weight of chlorine (Cl), 35.453 Acceptance criteria: NLT 90.0% and NMT 110.0% OTHER COMPONENTS CONTENT OF ALUMINUM Edetate disodium titrant: 37.2 mg/mL of edetate disodium Standardization of titrant: Transfer 2 g of aluminum wire to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) Ar = atomic weight of aluminum, 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 200 mg of Aluminum Dichlorohydrex Polyethylene Glycol to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acid-ammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). Use the aluminum content obtained to calculate the Aluminum/Chloride Atomic Ratio. CONTENT OF CHLORIDE Sample: 700 mg of Aluminum Dichlorohydrex Polyethylene Glycol Analysis: Transfer the Sample to a 250-mL beaker and add, with stirring, 100 mL of water and 10 mL of diluted nitric acid. Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). Use the chloride content thus obtained to calculate the Aluminum/Chloride Atomic Ratio. W Al
Aluminum Dichlorohydrex Propylene Glycol

(Comment on this Monograph)id=m2068=Aluminum Dichlorohydrex Propylene Glycol=A-Monos.pdf) Aly(OH)3y-zClz nH2O mC3H8O2 Aluminum chlorohydroxide propylene glycol complex; Aluminum hydroxychloride propylene glycol complex. DEFINITION Aluminum Dichlorohydrex Propylene Glycol consists of aluminum dichlorohydrate in which some of the waters of hydration have been replaced by propylene glycol. It encompasses a range of aluminum-to-chloride atomic ratios between 0.90:1 and 1.25:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum dichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements B. PROCEDURE Sample solution: Dissolve 0.5 g in 40 mL of water and, while mixing, adjust with 2.5 N sodium hydroxide to a pH of 9.55 0.05. Filter the suspension of precipitate thus obtained. Evaporate about 15 mL of the filtrate to about 1
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mL on a hot plate. Deposit the solution on a silver chloride disk. Standard solution: A similar preparation of propylene glycol Acceptance criteria: The IR absorption spectrum of a film of the Sample solution exhibits maxima only at the same wavelengths as that of a film of the Standard solution. ASSAY PROCEDURE Analysis Calculate the percentage of anhydrous aluminum dichlorohydrate in the Aluminum Dichlorohydrex Propylene Glycol: Result = Al%({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X) = percentage of aluminum as obtained in the test for Procedure 2: Content of Aluminum = atomic weight of aluminum, 26.98 Ar1 X = Aluminum/Chloride Atomic Ratio = molecular weight of the hydroxide ion (OH), Mr 17.01 = atomic weight of chlorine (Cl), 35.453 Ar2 Acceptance criteria: NLT 90.0%NMT 110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 700 mg of Aluminum Dichlorohydrex Propylene Glycol Analysis: Transfer the Sample to a 250-mL beaker and add 100 mL of water and 10 mL of diluted nitric acid with stirring. Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). [NOTEUse the results obtained to calculate the Aluminum/chloride atomic ratio.] CONTENT OF ALUMINUM Edetate disodium titrant: Prepare a solution with a concentration of 37.2 mg/mL of edetate disodium and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum (Al), 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 200 mg of Aluminum Dichlorohydrex Propylene Glycol to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 W Al%

mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). [NOTEUse the aluminum content thus obtained to calculate the Aluminum/chloride atomic ratio.] ALUMINUM/CHLORIDE ATOMIC RATIO Analysis: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of chloride found in the test for Content of chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 0.90:1 and 1.25:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Acceptance criteria: NMT 20 ppm LIMIT OF IRON 241 Standard solution: Pipet 2.0 mL of Standard Iron Solution, prepared as directed, into a 50-mL beaker Sample solution: 27 mg/mL Analysis: Pipet 5.0 mL of the Sample solution to a second 50-mL beaker. To each of the beakers add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). SPECIFIC TESTS PH 791 Sample solution: 15 in 100 (w/w) Acceptance criteria: Between 3.0 and 5.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum dichlorohydrate.
Aluminum Hydroxide Gel

(Comment on this Monograph)id=m2100=Aluminum Hydroxide Gel=A-Monos.pdf) Al(OH)3 Aluminum hydroxide [21645-51-2]. 78.00
DEFINITION Aluminum Hydroxide Gel is a suspension of amorphous aluminum hydroxide in which there is a partial substitution of carbonate for hydroxide. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of Al(OH)3. It may contain Peppermint Oil, Glycerin, Sorbitol, Sucrose, Saccharin, or other suitable flavors, and it may contain suitable antimicrobial agents. IDENTIFICATION A. PROCEDURE Sample: 1 g Analysis: Place the Sample in a flask equipped with a stopper and glass tubing, the tip of which is immersed in calcium hydroxide TS in a test tube. Add 5 mL of 3 N
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Analysis: Proceed as directed using a 20-mL portion of the Sample solution. Acceptance criteria: A 20-mL portion of the filtrate shows no more sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid [0.8%, based on the Al(OH)3 content]. ARSENIC, Method I 211 Sample solution: Amount of Aluminum Hydroxide Gel equivalent to 25 mg/mL of Al(OH)3 in 7 N sulfuric acid Standard solution: Prepare as directed in the test for Arsenic 211, except to prepare it to contain 5 g of arsenic instead of 3 g. Acceptance criteria: NMT 10 ppm, based on the Al(OH)3 content HEAVY METALS 231 Sample solution: Dissolve an amount of Aluminum Hydroxide Gel equivalent to 0.24 g of Al(OH)3 in 10 mL of 3 N hydrochloric with the aid of heat, filter, if necessary, and dilute with water to 25 mL. Acceptance criteria: NMT 83 ppm, based on the Al(OH)3 content SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62 Acceptance criteria: Its total aerobic microbial count does not exceed 100 cfu/mL, and it meets the requirements of the test for the absence of Escherichia coli. PH 791 Acceptance criteria: Between 5.5 and 8.0, determined potentiometrically ACID-NEUTRALIZING CAPACITY 301 Acceptance criteria: NLT 65.0% of the expected mEq value, calculated from the results of the above Assay, is obtained. [NOTEAl(OH)3 has an expected acid-neutralizing capacity value of 0.0385 mEq/mg.] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing.
hydrochloric acid to the flask, and immediately insert the stopper. Acceptance criteria: Gas evolves in the flask, and a precipitate is formed in the test tube. B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample: The solution remaining in the flask from Procedure A Acceptance criteria: Meets the requirements ASSAY PROCEDURE Edetate disodium titrant: Prepare a solution with a concentration of 18.6 mg/mL of edetate disodium and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/Ar V = weight aluminum in the portion of solution taken (mg) = atomic weight of aluminum (Al), 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer an amount of Aluminum Hydroxide Gel equivalent to 1.5 g of Al(OH)3 to a beaker, add 15 mL of hydrochloric acid, and heat gently until solution is complete. Cool, transfer to a 500-mL volumetric flask, and dilute to volume with water. Analysis: Pipet 20 mL of the Sample solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, then heat the solution near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, substituting 20 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.9 mg of Al(OH)3. Acceptance criteria: NLT 90.0% and NMT 110.0% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221 Sample: Amount of Aluminum Hydroxide Gel equivalent to 0.6 g of Al(OH)3 Analysis: Transfer the Sample to a porcelain dish, and add 0.1 mL of potassium chromate TS and 25 mL of water. Stir, and add 0.10 N silver nitrate until a faint, persistent pink color is obtained. Acceptance criteria: NMT 8.0 mL of 0.10 N silver nitrate is required [4.7%, based on the Al(OH)3 content]. CHLORIDE AND SULFATE, Sulfate 221 Sample solution: Add 5.0 mL of 3 N hydrochloric acid to an amount of Aluminum Hydroxide Gel equivalent to 0.3 g of Al(OH)3 and heat to dissolve the specimen under test. Cool, dilute with water to 250 mL, and filter if necessary. W
Dried Aluminum Hydroxide Gel

(Comment on this Monograph)id=m2110=Dried Aluminum Hydroxide Gel=A-Monos.pdf) Al(OH)3 Aluminum hydroxide [21645-51-2]. 78.00
DEFINITION Dried Aluminum Hydroxide Gel is an amorphous form of aluminum hydroxide in which there is a partial substitution of carbonate for hydroxide. It contains the equivalent of NLT 76.5% of Al(OH)3, and it may contain varying quantities of basic aluminum carbonate and bicarbonate. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample: Dissolve 5 mg in 10 mL of 3 N hydrochloric acid, with gentle warming. Acceptance criteria: The solution responds to the tests. ASSAY PROCEDURE Edetate disodium titrant: Prepare a solution with a concentration of 18.6 mg/mL of edetate disodium and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the
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reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/(Ar)(V) = weight aluminum in the portion of solution taken (mg) = atomic weight of aluminum (Al), 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer an amount of Dried Aluminum Hydroxide Gel equivalent to 2 g of Al(OH)3 to a beaker, add 15 mL of hydrochloric acid, and heat gently until solution is complete. Cool, transfer to a 500-mL volumetric flask, and dilute to volume with water. Analysis: Pipet 20 mL of the Sample solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, then heat the solution near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank titration, substituting 20 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant consumed is equivalent to 3.9 mg of Al(OH)3. Acceptance criteria: NLT 76.5% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221 Sample solution: Dissolve 1.0 g in 30 mL of 2 N nitric acid, heat to boiling, add water to make 100 mL, and filter. Analysis: Proceed as directed using a 5.0-mL portion of the Sample solution diluted with an equal volume of water. Acceptance criteria: A 5.0-mL portion of the Sample solution filtrate, diluted with an equal volume of water, shows no more chloride than corresponds to 0.60 mL of 0.20 N hydrochloric acid (0.85%). CHLORIDE AND SULFATE, Sulfate 221 Sample solution: Dissolve 330 mg in 15 mL of 3 N hydrochloric acid, heat to boiling, add water to make 250 mL, and filter. Analysis: Proceed as directed using a 25-mL portion of the Sample solution. Acceptance criteria: A 25-mL portion of the filtrate shows no more sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid (0.6%). ARSENIC, Method I 211 Sample solution: Dissolve 1.5 g in 80 mL of 7 N sulfuric acid, and dilute with water to 220 mL. Analysis: Proceed as directed using a 55-mL portion of the Sample solution, omitting the addition of 20 mL of 7 N sulfuric acid. Acceptance criteria: NMT 8 ppm HEAVY METALS 231 Sample solution: Dissolve 330 mg in 10 mL of 3 N hydrochloric with the aid of heat, filter, if necessary, and dilute with water to 25 mL. W

Acceptance criteria: NMT 60 ppm
SPECIFIC TESTS PH 791 Sample solution: 1 in 25 Acceptance criteria: NMT 10.0 ACID-NEUTRALIZING CAPACITY 301 Sample: 400 mg Analysis: Test as directed for Powders under Test Preparation. Acceptance criteria: NLT 25.0 mEq/g ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dried Aluminum Hydroxide Gel RS
Dried Aluminum Hydroxide Gel Capsules

(Comment on this Monograph)id=m2120=Dried Aluminum Hydroxide Gel Capsules=A-Monos.pdf) DEFINITION Dried Aluminum Hydroxide Gel Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of aluminum hydroxide [Al(OH)3]. IDENTIFICATION A. PROCEDURE Sample: Place a portion of Capsule contents, equivalent to about 500 mg of aluminum hydroxide. Analysis: Place in a flask equipped with a stopper and glass tubing, the tip of which is immersed in calcium hydroxide TS in a test tube. Add 10 mL of 3 N hydrochloric acid to the flask, and immediately insert the stopper: gas evolves in the flask and a precipitate is formed in the test tube. Acceptance Criteria: Gas evolves in the flask and a precipitate is formed in the test tube. B. IDENTIFICATION TESTSGENERAL, Aluminum 191: The solution remaining in the flask in Identification test A meets the requirements. ASSAY PROCEDURE Edetate disodium titrant: Prepare a solution with a concentration of 18.6 mg/mL of edetate disodium and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/(Ar)(V) W Ar = weight aluminum in the portion of solution taken (mg) = atomic weight of aluminum (Al), 26.98
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V
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add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/26.98V = weight of aluminum in the portion of solution taken (mg) V = volume of Edetate disodium titrant consumed (mL) Sample solution: Weigh and finely powder NLT 20 Tablets. Weigh a portion of the powder, equivalent to 1.2 g of aluminum hydroxide, add 15 mL of hydrochloric acid, and heat until dissolved. Dilute with water to about 100 mL, and filter quantitatively into a 500-mL volumetric flask, washing the filter with water, and dilute with water to volume. Analysis: Pipet 20 mL of the Sample solution into a 250-mL beaker, and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, then heat the solution near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 20 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701: 10 min, simulated gastric fluid TS being substituted for water in the test UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and NLT 55.0% of the expected mEq value, calculated from the labeled quantity of Al(OH)3, is obtained. Each mg of Al(OH)3 has an expected acid-neutralizing capacity value of 0.0385 mEq. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Tablets may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3. W
= volume of Edetate disodium titrant consumed (mL) Sample solution: Weigh the contents of NLT 20 Capsules. Transfer a weighed portion of the powder, equivalent to 1.2 g of aluminum hydroxide, to a beaker, add 15 mL of hydrochloric acid, and heat until dissolved. Dilute with water to about 100 mL, and filter quantitatively into a 500-mL volumetric flask, washing the filter with water, and dilute with water to volume. Analysis: Pipet 20 mL of Sample solution into a 250-mL beaker, and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, then heat the solution near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 20 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 3.9 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701: 10 min, simulated gastric fluid TS being substituted for water in the test UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and NLT 55.0% of the expected mEq value, calculated from the labeled quantity of Al(OH)3, is obtained. Each mg of Al(OH)3 has an expected acid-neutralizing capacity value of 0.0385 mEq. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Capsules may be labeled to state the aluminum hydroxide content in terms of the equivalent amount of dried aluminum hydroxide gel, on the basis that each mg of dried gel is equivalent to 0.765 mg of Al(OH)3.
Dried Aluminum Hydroxide Gel Tablets

(Comment on this Monograph)id=m2150=Dried Aluminum Hydroxide Gel Tablets=A-Monos.pdf) DEFINITION Dried Aluminum Hydroxide Gel Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aluminum hydroxide [Al(OH)3]. IDENTIFICATION A. PROCEDURE Sample: Finely ground Tablets, equivalent to 500 mg of aluminum hydroxide Analysis: Place in a flask equipped with a stopper and glass tubing, the tip of which is immersed in calcium hydroxide TS in a test tube. Add 5 mL of 3 N hydrochloric acid to the flask, and immediately insert the stopper. Acceptance criteria: Gas evolves in the flask, and a precipitate is formed in the test tube. B. IDENTIFICATION TESTSGENERAL, Aluminum 191: The solution remaining in the flask in Identification test A meets the requirements. ASSAY PROCEDURE Edetate disodium titrant: 18.6 mg/mL of edetate disodium in water Edetate disodium standardization: Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and
Aluminum Phosphate Gel

(Comment on this Monograph)id=m2270=Aluminum Phosphate Gel=A-Monos.pdf) Phosphoric acid, aluminum salt (1:1); Aluminum phosphate (1:1) [7784-30-7]. DEFINITION Aluminum Phosphate Gel is a water suspension containing NLT 4.0% and NMT 5.0% (w/w) of aluminum phosphate (AlPO4). It may contain sodium benzoate, benzoic acid, or other
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suitable agent, in an amount not exceeding 0.5%, as a preservative. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum 191: A solution of it in hydrochloric acid meets the requirements. B. IDENTIFICATION TESTSGENERAL, Phosphate 191: A solution of it in 2 N nitric acid meets the requirements. ASSAY PROCEDURE Sample solution: To 20 g of Gel, in a 100-mL volumetric flask, add nitric acid to dissolve, dilute with water to volume. Analysis: Transfer 10.0 mL of Sample solution to a 400-mL beaker, dilute with water to 100 mL, heat to 60, add an excess of ammonium molybdate TS, and maintain at 50 for 30 min. Filter, and wash the precipitate with dilute nitric acid (1 in 36), then with potassium nitrate solution (1 in 100) until the last portion of the filtrate is not acid to litmus paper. Dissolve the precipitate in 50.0 mL of 0.5 N sodium hydroxide VS, add phenolphthalein TS, and titrate the excess sodium hydroxide with 0.5 N sulfuric acid VS. Each mL of 0.5 N sodium hydroxide is equivalent to 2.651 mg of AlPO4. Acceptance criteria: 4.0%5.0% (w/w) IMPURITIES Inorganic Impurities CHLORIDE: Sample Solution: Transfer 25 g to a beaker with the aid of 50 mL of water, add 5 mL of nitric acid, then add, with stirring, 30.0 mL of 0.1 N silver nitrate VS. Warm on a steam bath for 30 min, filter, and wash the precipitate with water acidified with nitric acid. Analysis: To the filtrate add ferric ammonium sulfate TS, and titrate the excess silver nitrate with 0.1 N ammonium thiocyanate VS. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Cl. Acceptance criteria: NMT 0.16% CHLORIDE AND SULFATE, Sulfate 221 Sample solution: Add 10 mL of 3 N hydrochloric acid to 10 g of Gel, and heat to boiling. Cool, dilute with water to 250 mL, and filter, if necessary. Acceptance criteria: A 10-mL portion of the solution shows no more sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid: NMT 0.05%. ARSENIC, Method I 211 Sample solution: Dissolve 5.0 g of Gel in the smallest necessary volume of 3 N hydrochloric acid. Acceptance criteria: NMT 0.6 ppm HEAVY METALS, Method I 231 [NOTEProceed as directed in the chapter, except to make the following modifications.] Standard solution: Into a 50-mL color-comparison tube pipet 4.0 mL of Standard Lead Solution, dilute with water to 25 mL, adjust with 6 N ammonium hydroxide to a pH between 1.9 and 2.1, and dilute with water to 40 mL. Sample solution: Dissolve 8 g in 5 mL of 3 N hydrochloric acid, warming if necessary, dilute with water to 25 mL, and adjust with 6 N ammonium hydroxide to a pH between 1.9 and 2.1. Transfer to a 50-mL color-comparison tube, and dilute with water to 40 mL. Monitor solution: Into a 50-mL color-comparison tube place 25 mL of the Sample solution, add 4.0 mL of Standard Lead Solution, adjust with 1 N acetic acid or 6 N ammonium hydroxide to a pH between 1.9 and 2.1, and dilute with water to 40 mL. Analysis: Proceed as directed in the chapter, except to omit the addition of 2 mL of pH 3.5 Acetate Buffer.

Acceptance criteria: NMT 5 ppm
SPECIFIC TESTS PH 791: 6.07.2 SOLUBLE PHOSPHATE: Filter 20 g and wash the residue with 30 mL of water. Add to the filtrate 2 mL of nitric acid, heat to 60, and add 20 mL of ammonium molybdate TS. Heat at 50 for 30 min, filter, wash the precipitate with dilute nitric acid (1 in 36), then wash with potassium nitrate solution (1 in 100) until the last portion of the filtrate is not acid to litmus paper. Dissolve the precipitate in 50.0 mL of 0.5 N sodium hydroxide VS, add phenolphthalein TS, and titrate the excess alkali with 0.5 N hydrochloric acid VS. Each mL of 0.5 N sodium hydroxide is equivalent to 2.065 mg of PO4. Acceptance criteria: Soluble phosphate, calculated as PO4, NMT 0.30% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Aluminum Sesquichlorohydrate
(Comment on this Monograph)id=m2285=Aluminum Sesquichlorohydrate=A-Monos.pdf) Aly(OH)3y-zClz nH2O Aluminum chlorohydroxide; Aluminum hydroxychloride [11097-68-0]. DEFINITION Aluminum Sesquichlorohydrate consists of complex basic aluminum chloride that is polymeric and loosely hydrated and encompasses a range of aluminum-to-chloride atomic ratios between 1.26:1 and 1.90:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum sesquichlorohydrate. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191: A 100 mg/mL solution meets the requirements of the tests for Aluminum and for Chloride. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum sesquichlorohydrate in the Aluminum Sesquichlorohydrate: Result = Al ({Ar1x + [Mr(3x -1)] + Ar2}/Ar1x) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 x = aluminum/chloride atomic ratio found in the test for Aluminum/Chloride Atomic Ratio = molecular weight of the hydroxide anion (OH), Mr 17.01 = atomic weight of chlorine (Cl), 35.453 Ar2 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 700 mg of Aluminum Sesquichlorohydrate to a 250-mL beaker. Add 100 mL of water and 10 mL of diluted nitric acid with stirring. Analysis: Titrate with 0.1 N silver nitrate VS using a glass silver-silver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). [NOTEUse the chloride content thus obtained to calculate the Aluminum/chloride atomic ratio.] Al
138

SPECIFIC TESTS PH 791: 3.05.0, in a solution [15 in 100 (w/w)]
USP 32
CONTENT OF ALUMINUM Edetate disodium titrant: Prepare a solution with a concentration of 37.2 mg/mL of edetate disodium and standarize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: W/Ar V = weight of aluminum in the portion of Solution taken (mg) Ar = atomic weight of aluminum, 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 200 mg of Aluminum Sesquichlorohydrate to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). [NOTEUse the aluminum content thus obtained to calculate the Aluminum/chloride atomic ratio.] ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of chloride found in the test for Content of chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 1.26:1 and 1.90:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm HEAVY METALS, Method I 231: NMT 20 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Sesquichlorohydrate in water. Transfer 5.0 mL of this solution to a 50 mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). W
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum sesquichlorohydrate.
Aluminum Sesquichlorohydrate Solution

(Comment on this Monograph)id=m2286=Aluminum Sesquichlorohydrate Solution=A-Monos.pdf) DEFINITION Aluminum Sesquichlorohydrate Solution consists of complex basic aluminum chloride that is polymeric and encompasses a range of aluminum-to-chloride atomic ratios between 1.26:1 and 1.90:1. The following solvents may be used: water, propylene glycol, dipropylene glycol, or alcohol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum sesquichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191: A solution containing the equivalent of 100 mg of anhydrous aluminum sesquichlorohydrate/mL meets the requirements of the tests for Aluminum and for Chloride. B. IDENTIFICATION OF PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL of Solution in isopropyl alcohol, and filter. Evaporate the filtrate to 1 mL on a steam bath. Acceptance criteria: The IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. IDENTIFICATION OF DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL of Solution in isopropyl alcohol, and filter. Evaporate the filtrate to 1 mL on a steam bath. Acceptance criteria: The IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. D. IDENTIFICATION OF ALCOHOL (where stated on the label) Sample solution: Mix 5 drops of Solution in a small beaker with 1 mL of potassium permanganate solution (1 in 100) and 5 drops of 2 N sulfuric acid, and cover the beaker immediately with filter paper moistened with a freshly prepared solution of 0.1 g of sodium nitroferricyanide and 0.25 g of piperazine in 5 mL of water. Acceptance criteria: An intense blue color is produced on the filter paper, the color becoming paler after a few min. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum sesquichlorohydrate in the Solution: Result = Al({Ar1x + [Mr(3x -1)] + Ar2}/Ar1x) Al Ar1 x Mr Ar2 = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 = aluminum/chloride atomic ratio = molecular weight of the hydroxide anion, 17.01 = atomic weight of chlorine, 35.453
USP 32

Acceptance criteria: NMT 10 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 53 mg/mL of Aluminum Sesquichlorohydrate Solution. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (75 ppm). SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label Solution to state the solvent used and the claimed concentration of anhydrous aluminum sesquichlorohydrate contained therein.
OTHER COMPONENTS CONTENT OF CHLORIDE Sample solution: 1.4 g of Aluminum Sesquichlorohydrate Solution to a 250-mL beaker, add 100 mL of water and 10 mL of diluted nitric acid with stirring. Analysis: Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). [NOTEUse the chloride content thus obtained to calculate the Aluminum/chloride atomic ratio.] CONTENT OF ALUMINUM Edetate disodium titrant: Prepare a solution with a concentration of 37.2 mg/mL of edetate disodium, and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV = weight, of aluminum in the portion of solution taken (mg) Ar = atomic weight of aluminum, 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 400 mg of Aluminum Sesquichlorohydrate Solution, to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). [NOTEUse the aluminum content thus obtained to calculate the Aluminum/chloride atomic ratio.] ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of chloride found in the test for Content of chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 1.26:1 and 1.90:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Sample solution: Prepare as directed for Test Preparation in the chapter, using a weighed quantity of the Solution. Acceptance criteria: NMT 2 ppm HEAVY METALS, Method I 231 Sample solution: Prepare as directed for Test Preparatiion in the chapter, using a weighed quantity of the Solution. W
Aluminum Sesquichlorohydrex Polyethylene Glycol

(Comment on this Monograph)id=m2290=Aluminum Sesquichlorohydrex Polyethylene Glycol=A-Monos.pdf) Aly(OH)3y-zClz nH2O mH(OCH2CH2)nOH Aluminum chlorohydroxide polyethylene glycol complex; Aluminum hydroxychloride polyethylene glycol complex. DEFINITION Aluminum Sesquichlorohydrex Polyethylene Glycol consists of aluminum sesquichlorohydrate in which some of the waters of hydration have been replaced by polyethylene glycol. It encompasses a range of aluminum-to-chloride atomic ratios between 1.26:1 and 1.90:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum sesquichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191: A 100 mg/mL solution meets the requirements. B. INFRARED ABSORPTION 197F Sample solution: Dissolve 0.5 g in about 40 mL of water, and while mixing, adjust with 2.5 N sodium hydroxide to a pH of 9.55 0.05. Filter the suspension of precipitate thus obtained. Evaporate 15 mL of the filtrate to 1 mL on a hot plate. Deposit this solution on a silver chloride disk. Standard solution: A similar preparation of polyethylene glycol ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum sesquichlorohydrex in the Aluminum Sesquichlorohydrex Polyethylene Glycol: Result = Al%({Ar1X + [Mr(3X 1)] + Ar2}/Ar1X) Al% Ar1 X = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 = aluminum/chloride atomic ratio found in the test for Aluminum/chloride atomic ratio
140
Mr

Acceptance criteria: 1.90:1.
USP 32
The ratio is between 1.26:1 and
= molecular weight of the hydroxide anion (OH), 17.01 = atomic weight of chlorine (Cl), 35.453 Ar2 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample solution: 700 mg of Aluminum Sesquichlorohydrex Polyethylene Glycol to a 250-mL beaker. Add 100 mL of water and 10 mL of diluted nitric acid with stirring. Analysis: Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system, determining the endpoint potentiometrically. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). [NOTEUse the chloride content thus obtained to calculate the Aluminum/chloride atomic ratio.] CONTENT OF ALUMINUM Edetate disodium titrant: Prepare a solution with a concentration of 37.2 mg/mL of edetate disodium in water, and standardize as follows. Weigh 2 g of aluminum wire. Transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/(Ar)(V) = weight of aluminum in the portion of Solution taken (mg) Ar = atomic weight of aluminum, 26.98 V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer about 200 mg of Aluminum Sesquichlorohydrex Polyethylene Glycol to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the Sample solution, add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (Al). [NOTEUse the aluminum content thus obtained to calculate the Aluminum/chloride atomic ratio.] ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of chloride found in the test for Content of chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. W
IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm HEAVY METALS, Method I 231: NMT 20 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, and prepare as directed under Iron 241, to a 50mL beaker. Sample solution: 27 mg/mL of Aluminum Sesquichlorohydrex Polyethylene Glycol in water. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed in Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). SPECIFIC TESTS PH 791: 3.05.0, in a solution of 15 in 100 (w/w) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum sesquichlorohydrate.
Aluminum Sesquichlorohydrex Propylene Glycol

(Comment on this Monograph)id=m2293=Aluminum Sesquichlorohydrex Propylene Glycol=A-Monos.pdf) Aly(OH)3yzClz nH2O mC3H8O2 Aluminum chlorohydroxide propylene glycol complex; Aluminum hydroxychloride propylene glycol complex. DEFINITION Aluminum Sesquichlorohydrex Propylene Glycol consists of aluminum sesquichlorohydrate in which some of the waters of hydration have been replaced by propylene glycol. It encompasses a range of aluminum-to-chloride atomic ratios between 1.26:1 and 1.90:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum sesquichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum and Chloride 191: A 100 mg/mL solution meets the requirements. B. INFRARED ABSORPTION 197F: Sample solution: Dissolve 0.5 g in about 40 mL of water, and while mixing adjust with 2.5 N sodium hydroxide to a pH of 9.55 0.05. Filter the suspension of precipitate thus obtained. Evaporate 15 mL of the filtrate to 1 mL on a hot plate. Deposit this solution on a silver chloride disk. Standard solution: A similar preparation of propylene glycol
USP 32
ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum sesquichlorohydrate in the Aluminum Sesquichlorohydrex Propylene Glycol: Result = Al({Ar1X + [Mr(3X -1)] + Ar2}/Ar1X) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 X = aluminum/chloride atomic ratio found in the test for Aluminum/chloride atomic ratio = molecular weight of the hydroxide anion (OH), Mr 17.01 = atomic weight of chlorine (Cl), 35.453 Ar2 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample solution: 700 mg of Aluminum Sesquichlorohydrex Propylene Glycol to a 250-mL beaker. Add 100 mL of water and 10 mL of diluted nitric acid with stirring. Analysis: Titrate with 0.1 N silver nitrate VS using a glass silversilver chloride electrode and a silver billet electrode system. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride (Cl). [NOTEUse the chloride content thus obtained to calculate the Aluminum/chloride atomic ratio.] CONTENT OF ALUMINUM Edetate disodium titrant: Prepare a solution with a concentration of 37.2 mg/mL of edetate disodium, and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV = weight of aluminum in the portion of Solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer 200 mg of Aluminum Sesquichlorohydrex Propylene Glycol to a 250-mL beaker, add 20 mL of water and 5 mL of hydrochloric acid, boil on a hot plate for NLT 5 min, and allow to cool. Analysis: To the 25 mL of Sample solution add 25.0 mL of Edetate disodium titrant, and adjust with 2.5 N ammonium hydroxide or 1 N acetic acid to a pH of 4.7 0.1. Add 20 mL of acetic acidammonium acetate buffer TS, 50 mL of alcohol, and 5 mL of dithizone TS. The pH of this solution should be 4.7 0.1. Titrate with 0.1 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank titration, and make any necessary correction. Each mL of 0.1 M Edetate disodium titrant consumed is equivalent to 2.698 mg of aluminum (AL). [NOTEUse the aluminum content thus obtained to calculate the Aluminum/chloride atomic ratio.] ALUMINUM/CHLORIDE ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by W Al

the percentage of chloride found in the test for Content of chloride, and multiply by 35.453/26.98, in which 35.453 and 26.98 are the atomic weights of chlorine and aluminum, respectively. Acceptance criteria: The ratio is between 1.26:1 and 1.90:1. IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm HEAVY METALS, Method I 231: NMT 20 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Sesquichlorohydrex Propylene Glycol. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 3 to 5 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (150 ppm). SPECIFIC TESTS PH 791: 3.05.0, in a solution [15 in 100 (w/w)] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum sesquichlorohydrate.
Aluminum Subacetate Topical Solution

(Comment on this Monograph)id=m2300=Aluminum Subacetate Topical Solution=A-Monos.pdf)
C4H7AlO5 Aluminum, bis(acetato-O)hydroxy-; Bis(acetato)hydroxyaluminum; Basic aluminum acetate [142-03-0; 8000-61-1].
162.08
DEFINITION Aluminum Subacetate Topical Solution yields, from each 100 mL, NLT 2.30 g and NMT 2.60 g of Al2O3, and NLT 5.43 g and NMT 6.13 g of C2H4O2. It may be stabilized by the addition of NMT 0.9% of boric acid. Aluminum Subacetate Topical Solution may be prepared as follows:
Aluminum Sulfate Acetic Acid Calcium Carbonate Purified Water To make 145 g 160 mL 70 g A sufficient quantity 1000 mL
142

Acceptance criteria: 5.43 g6.13 g of C2H4O2
USP 32
Dissolve the aluminum sulfate in 600 mL of cold water, filter the solution, and add the calcium carbonate gradually, in several portions, with constant stirring. Then slowly add the acetic acid, mix, and set the mixture aside for 24 h. Filter the product with the aid of vacuum if necessary, returning the first portion of the filtrate to the funnel. Wash the magma on the filter with small portions of cold water, until the total filtrate measures 1000 mL. IDENTIFICATION GENERAL, Aluminum and Sulfate IDENTIFICATION TESTS 191: Meets the requirements ASSAY ALUMINUM OXIDE Edetate disodium titrant: Prepare a solution with a concentration of 18.6 mg/mL of edetate disodium and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary connection. Calculate the molarity of the solution taken by the formula: Result = W/Ar V = weight in mg, of aluminum in the portion of Solution = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: Pipet 20 mL of Topical Solution into a 250-mL volumetric flask, add 5 mL of hydrochloric acid, and dilute with water to volume. Analysis: Pipet 25 mL of Sample solution into a 250-mL beaker, and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, then heat the solution near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting water for the sample. Each mL of 0.05 M Edetate disodium titrant is equivalent to 2.549 mg of Al2O3 Acceptance criteria: 2.30 2.60 g of Al2O3 ACETIC ACID Sample solution: Pipet 20 mL of Topical Solution into a Kjeldahl flask containing a mixture of 20 mL of phosphoric acid and 150 mL of water. Analysis: Connect the flask to a condenser, the delivery tube from which dips beneath the surface of 50.0 mL of 0.5 N sodium hydroxide VS contained in a receiving flask. Distill 160 mL, then remove the delivery tube from below the surface of the liquid, allow the distilling flask to cool, add 50 mL of water, and distill an additional 40 to 45 mL into the receiving flask. Add phenolphthalein TS to the distillate, and titrate the excess 0.5 N sodium hydroxide VS with 0.5 N sulfuric acid VS. Each mL of 0.5 N sodium hydroxide is equivalent to 30.03 mg of C2H4O2 W
SPECIFIC TESTS PH 791: 3.84.6 LIMIT OF BORIC ACID Sample solution: Pipet 25 mL of Aluminum Subacetate Topical Solution into 75 mL of water in a conical flask. Analysis: To 100 mL of Sample solution, add 3 mL of phenolphthalein TS, then add 0.5 N sodium hydroxide VS from a buret until a faint pink color is obtained. Heat to boiling, and again neutralize. Add 150 mL of glycerin to the neutralized solution, and titrate with 0.5 N sodium hydroxide VS. Perform a blank determination in a similar manner. From the volume of 0.5 N sodium hydroxide VS used after the addition of the glycerin, subtract the volume used in the blank. Each mL of 0.5 N sodium hydroxide is equivalent to 30.92 mg of H3BO3 Acceptance criteria: NMT 0.6% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Aluminum Sulfate
(Comment on this Monograph)id=m2350=Aluminum Sulfate=AMonos.pdf) Al2(SO4)3 xH2O (anhydrous) Sulfuric acid, aluminum salt (3:2), hydrate; Aluminum sulfate (2:3) hydrate [17927-65-0]. Anhydrous [10043-01-3].
342.15
DEFINITION Aluminum Sulfate contains NLT 54.0% and NMT 59.0% of Al2(SO4)3. It contains a varying amount of water of crystallization. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Aluminum and Sulfate 191: A 100 mg/mL solution meets the requirements of the tests for Aluminum and for Sulfate. ASSAY PROCEDURE Edetate disodium titrant: Prepare a solution with a concentration of 18.6 mg/mL of edetate disodium and standardize as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/ArV W Ar = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98
USP 32
= volume of Edetate disodium titrant consumed (mL) Sample solution: 30.0 mg/mL of Aluminum Sulfate Analysis: Pipet 10 mL of the Sample solution into a 250-mL beaker, and add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, then heat the solution near the boiling point for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate the solution with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting water for the sample, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 8.554 mg of Al2(SO4)3. Acceptance criteria: 54.0%59.0% IMPURITIES Inorganic Impurities IRON 241 Sample solution: To 20 mL of a solution (1 in 150) add 0.3 mL of potassium ferrocyanide TS. Acceptance criteria: No blue color is produced immediately. HEAVY METALS 231 Sample solution: Dissolve 1.0 g in 2 mL of 1 N acetic acid, and dilute with water to 25 mL. Acceptance criteria: NMT 20 ppm SPECIFIC TESTS PH 791: NLT 2.9, in a solution (1 in 20) WATER DETERMINATION, Method I 921: 41.0%46.0% LIMIT OF ALKALIES AND ALKALINE EARTHS: To a boiling solution of 1.0 g in 150 mL of water add a few drops of methyl red TS, and then add 6 N ammonium hydroxide just until the color of the solution changes to a distinct yellow. Add hot water to restore the volume to 150 mL, and filter while hot. Evaporate 75 mL of the filtrate to dryness, and ignite to constant weight. Acceptance criteria: NMT 2 mg of residue remains (0.4%). LIMIT OF AMMONIUM SALTS: Heat 1 g with 10 mL of 1 N sodium hydroxide on a steam bath for 1 min. Acceptance criteria: The odor of ammonia is not perceptible. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. V

B. IDENTIFICATION TESTSGENERAL, Sulfate and Calcium 191: Suspend 2 g of sample in 50 mL of, and filter. The filtrate meets the requirements. ASSAY ALUMINUM SULFATE Sample solution: Transfer 10 g of Aluminum Sulfate and Calcium Acetate for Topical Solution to a 1000-mL volumetric flask. Add 100 mL of 1.2 M hydrochloric acid and 250 mL of water. Heat on a steam bath or hot plate until dissolved. Cool, dilute with water to volume. [NOTERetain a portion of this Sample solution for the Assay for Calcium acetate.] Analysis: Transfer a 5.0-mL aliquot of the Sample solution to a 250-mL conical flask. Add 40.0 mL of 0.01 M edetate disodium VS and 20 mL of acetic acidammonium acetate buffer TS. Add 50 mL of alcohol and 2 mL of dithizone TS. [NOTEFollow the given order of addition.] Titrate with 0.02 M zinc sulfate VS until the color changes from green-violet to clear rose-pink. Perform a blank titration, substituting 5.0 mL of water for the Sample solution. Each mL of 0.01 M edetate disodium is equivalent to 2.972 mg of Al2(SO4)3 14H2O. Calculate the percentage of Al2(SO4)3 14H2O taken: Result = [(1000)(100)F M(VB VU)]/5.0 MTW = dilution factor, 1000/5.0 = conversion factor to percentage = conversion factor (2.972 mg of sample/mL of 0.01 M edetate disodium) M = actual molarity of the titrant = blank titration volume (mL) VB = sample titration volume (mL) VU = theoretical molarity of the titrant, 0.02 MT W = weight of the sample (mg) CALCIUM ACETATE Analysis: Transfer a 5.0-mL aliquot of the Sample solution retained from the Assay for aluminum sulfate to a 250-mL conical flask. [NOTEFollow the given order of addition.] Add 1 to 2 mL of 50% triethanolamine to mask the aluminum, mix well. Add 100 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Titrate the solution with 0.01 M edetate disodium VS. The indicator will change from purple to a clear blue color at the endpoint. Each mL of 0.01 M edetate disodium is equivalent to 1.762 mg of C4H6CaO4 H2O. Calculate the percentage of C4H6CaO4 H2O taken: Result = [(1000)(100)VUF M]/5.0 MTW dilution factor, 1000/5.0 conversion factor to percentage sample titration volume (mL) conversion factor (1.762 mg of sample/mL of 0.01 M edetate disodium) M = actual molarity of the titrant MT = theoretical molarity of the titrant. 0.01 W = weight of the sample (mg) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.04.8, in a solution (1 in 200) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-unit containers, and protect from excessive heat. D 100 VU F = = = = D 100 F
Aluminum Sulfate and Calcium Acetate for Topical Solution

(Comment on this Monograph)id=m1370=Aluminum Sulfate and Calcium Acetate for Topical Solution=A-Monos.pdf) DEFINITION Aluminum Sulfate and Calcium Acetate for Topical Solution contains NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum sulfate tetradecahydrate [Al2(SO4)3 14H2O] and calcium acetate monohydrate (C4H6CaO4 H2O). IDENTIFICATION A. Place approximately 0.25 g of Aluminum Sulfate and Calcium Acetate for Topical Solution in a test tube. Add 10 mL of water and 0.25 g of calcium carbonate. Heat on a steam bath for 10 min, and filter. Add 3 to 4 drops of ferric chloride TS to the filtrate. A reddish-brown color or precipitate indicates acetate. [NOTEAfter the addition of the ferric chloride TS, the solution may be heated for 1 min to speed the reaction.]
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Aluminum Sulfate and Calcium Acetate Tablets for Topical Solution

(Comment on this Monograph)id=m2353=Aluminum Sulfate and Calcium Acetate Tablets for Topical Solution=A-Monos.pdf) DEFINITION Aluminum Sulfate and Calcium Acetate Tablets for Topical Solution contain NLT 90.0% and NMT 110.0% of the labeled amounts of aluminum sulfate tetradecahydrate [Al2(SO4)3 14H2O] and calcium acetate monohydrate (C4H6CaO4 H2O). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: 40 mg/mL of ground Tablet powder in water, and filter Analysis: Mix 2 mL of the Sample solution with 2 mL of water and 2 drops of 3 N hydrochloric acid. Acceptance criteria: The solution responds to the ammonium hydroxide test. [NOTERetain the remaining filtrate for Identification test B.] B. IDENTIFICATION TESTSGENERAL, Sulfate and Calcium 191: A portion of the filtrate retained from Identification test A responds to the tests for Sulfate and for Calcium. ASSAY ALUMINUM SULFATE Sample solution: Finely powder and mix not fewer than 20 Tablets. Weigh a portion of the powder, equivalent to 2.8 g of aluminum sulfate, and transfer to a 1000-mL volumetric flask. Add 100 mL of 1.2 N hydrochloric acid and 100 mL of water, and heat on a steam bath, with occasional swirling, to dissolve the powder. Allow the solution to cool, dilute with water to volume. [NOTERetain a portion of this Sample solution for the Assay for Calcium Acetate.] Analysis: Transfer 25.0 mL of the Sample solution to a 250mL conical flask. Add 40.0 mL of 0.01 M edetate disodium VS and 20 mL of acetic acidammonium acetate buffer TS, and mix by swirling. Add 50 mL of alcohol and 2 mL of dithizone TS, and titrate with 0.02 M zinc sulfate VS until the color changes from green-violet to a clear rose-pink. Perform a blank determination, substituting 25 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.01 M edetate disodium is equivalent to 2.972 mg of Al2(SO4)3 14H2O. Acceptance criteria: 90.0%110.0% CALCIUM ACETATE Analysis: Transfer 20.0 mL of the Sample solution retained from the Assay for Aluminum Sulfate to a 125-mL conical flask. With constant stirring, add in the order named, 0.5 mL of trolamine, 10 mL of ammoniaammonium chloride buffer TS, and 3 drops of a solution prepared by dissolving 500 mg of eriochrome black T trituration in 10 mL of methanol, and titrate with 0.01 M edetate disodium VS to a violet endpoint. Each mL of 0.01 M edetate disodium is equivalent to 1.762 mg of C4H6CaO4 H2O. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701: 10 min UNIFORMITY OF DOSAGE UNITS 905: for Weight Variation
Aluminum Zirconium Octachlorohydrate

(Comment on this Monograph)id=m2355=Aluminum Zirconium Octachlorohydrate=A-Monos.pdf) AlyZr(OH)3y+4-xClx nH2O DEFINITION Aluminum Zirconium Octachlorohydrate is a polymeric, loosely hydrated complex of basic aluminum zirconium chloride that encompasses a range of aluminum-to-zirconium atomic ratios between 6.0:1 and 10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum zirconium octachlorohydrate. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of 100 mg/mL of anhydrous aluminum zirconium octachlorohydrate meets the requirements of the test for Chloride. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium octachlorohydrate in the Aluminum Zirconium Octachlorohydrate: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio found in the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Octachlorohydrate to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Octachlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and, while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] Al%
SPECIFIC TESTS PH 791: Between 4.0 and 4.8, in a solution of 2 g of ground Tablet powder in 500 mL of water LOSS ON DRYING 731: Dry ground Tablet powder at 150 for 15 min: it loses NMT 18% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid excessive heat.
USP 32
CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Octachlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Octachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium octachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum, as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium Ar2 = atomic weight of zirconium corrected for 2% hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 1.5:10.9:1 SPECIFIC TESTS PH 791: 3.05.0, in a solution (15 in 100) IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Al% = = = =

Sample solution: 27 mg/mL of Aluminum Zirconium Octachlorohydrate. Transfer 5.0 mL of this solution to a 50mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 150 ppm) HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. Acceptance criteria: NMT 20 ppm ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum zirconium octachlorohydrate.
Aluminum Zirconium Octachlorohydrate Solution

(Comment on this Monograph)id=m2356=Aluminum Zirconium Octachlorohydrate Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Octachlorohydrate Solution consists of complex basic aluminum chloride that is polymeric and encompasses a range of aluminum-to-zirconium atomic ratios between 6.0:1 and 10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium octachlorohydrate.
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CONTENT OF ALUMINUM Sample: 0.3 g of Aluminum Zirconium Octachlorohydrate Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Octachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of Aluminum Zirconium Octachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium Ar2 = atomic weight of zirconium corrected for 2% hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 1.5:10.9:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: Prepare the Sample solution using a weighed quantity of the Solution. Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Al% = = = =
IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of 100 mg/mL of anhydrous aluminum zirconium octachlorohydrate meets the requirements of the test for Chloride. B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium octachlorohydrate in the Solution taken: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio found in the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% SPECIFIC TESTS CONTENT OF CHLORIDE Sample: 500 mg of Aluminum Zirconium Octachlorohydrate Solution to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Aluminum Zirconium Octachlorohydrate Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] Al%
USP 32
Sample solution: 53 mg/mL of Aluminum Zirconium Octachlorohydrate Solution Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 75 ppm). HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using a weighed quantity of Solution. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for the Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Solution to state the solvent used and the claimed concentration of anhydrous aluminum zirconium octachlorohydrate.

10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum zirconium octachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100 mg/mL solution meets the requirements. B. PROCEDURE Sample solution: 25 mg/mL in water Analysis: Heat to boiling on a hot plate, and add 60 mg of ninhydrin to 20 mL of Sample solution. Acceptance criteria: A deep violet color immediately develops. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium octachlorohydrate in the Aluminum Zirconium Octachlorohydrate Gly in the solution taken: Result = Al% ({Ar1y + Ar2 + Mr1[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/ Ar1y) = percentage of aluminum from the test for Content of Aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/Zirconium Atomic Ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr1 17.01 z = (aluminum plus zirconium)/chloride atomic ratio from the test for (Aluminum plus Zirconium)/Chloride Atomic Ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Octachlorohydrate Gly to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). Use the result obtained to calculate the (Aluminum plus Zirconium)/Chloride Atomic Ratio. CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Octachlorohydrate Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and, while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). Use the result obtained to calculate the Aluminum/Zirconium Atomic Ratio and the (Aluminum plus Zirconium)/Chloride Atomic Ratio. CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Octachlorohydrate Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL Al%
Aluminum Zirconium Octachlorohydrex Gly

(Comment on this Monograph)id=m2358=Aluminum Zirconium Octachlorohydrex Gly=A-Monos.pdf) DEFINITION Aluminum Zirconium Octachlorohydrex Gly is a derivative of Aluminum Zirconium Octachlorohydrate in which some of the water molecules have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium atomic ratios between 6.0:1 and
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HEAVY METALS, Method I 231 [NOTEProceed as directed in the chapter, except to use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid prior to adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 20 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (15 in 100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the form of glycine used and the claimed content of anhydrous aluminum zirconium octachlorohydrate.
of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Octachlorohydrate Gly: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of Aluminum Zirconium Octachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum: the ratio is between 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Analysis: Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al %/Ar1) + (Zr/Ar2)]/(Cl/Ar3) = percentage of aluminum from the test for Content of aluminum Zr = percentage of zirconium from the test for Content of zirconium Cl = percentage of chloride from the test for Content of chloride Ar1 = atomic weight of aluminum, 26.98 Ar2 = atomic weight of zirconium corrected for 2% hafnium content, 92.97 = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 1.5:10.9:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Zirconium Octachlorohydrate Gly. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool, add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (NMT 150 ppm). Al % = = = =
Aluminum Zirconium Octachlorohydrex Gly Solution

(Comment on this Monograph)id=m2359=Aluminum Zirconium Octachlorohydrex Gly Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Octachlorohydrex Gly Solution is a solution of Aluminum Zirconium Octachlorohydrate in which some of the waters of hydration have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium ratios between 6.0:1 and 10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium octachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of 100 mg/mL of anhydrous aluminum zirconium octachlorohydrate meets the requirements.
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B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. D. GLYCINE Sample solution: 50 mg/mL in isopropyl alcohol, and filter Analysis: Heat to boiling on a hot plate, and add 60 mg of ninhydrin for 20 mL of Sample solution. Acceptance criteria: A deep violet color develops immediately. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium octachlorohydrate gly in the Solution taken: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio from the test for (Aluminum plus zirconium)/chloride atomic ratio Ar3 = atomic weight of chlorine (Cl), 35.453 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 500 mg of Aluminum Zirconium Octachlorohydrate Gly Solution in a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS, using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Aluminum Zirconium Octachlorohydrate Gly Solution in a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] Al%

CONTENT OF ALUMINUM Sample: 0.30 g of Aluminum Zirconium Octachlorohydrate Gly Solution in a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Octachlorohydrate Gly: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of Aluminum Zirconium Octachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Analysis: Calculate: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium = atomic weight of zirconium, corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 1.5:10.9:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: Prepare the Sample solution using a weighed quantity of the solution. Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 54 mg/mL of Aluminum Zirconium Octachlorohydrate Gly Solution Transfer 5.0 mL of this solution to a 50-mL beaker. Al% = = = =
150

amount of anhydrous aluminum zirconium pentachlorohydrate. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Chloride 191: mg/mL solution meets the requirements
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Analysis: To each of the beakers containing the Standard solution and the Sample solution add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 75 ppm) HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using an accurately weighed quantity of solution. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm The color of the solution from the Sample solution is not darker than that of the solution made from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution made from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Solution to state the solvent and form of glycine used and the claimed concentration of anhydrous aluminum zirconium octachlorohydrate.
A 100
ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium pentachlorohydrate in the Aluminum Zirconium Pentachlorohydrate: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio from the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Pentachlorohydrate to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Pentachlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Pentachlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink Al%
Aluminum Zirconium Pentachlorohydrate

(Comment on this Monograph)id=m2360=Aluminum Zirconium Pentachlorohydrate=A-Monos.pdf) AlyZr(OH)3y+4-xClx nH2O DEFINITION Aluminum Zirconium Pentachlorohydrate is a polymeric, loosely hydrated complex of basic aluminum zirconium chloride that encompasses a range of aluminum-to-zirconium atomic ratios between 6.0:1 and 10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. It contains NLT 90.0% and NMT 110.0% of the labeled
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color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Pentachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of Aluminum Zirconium Octachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium Ar2 = atomic weight of zirconium corrected for 2% hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 2.1:11.51:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Zirconium Pentachlorohydrate. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 150 ppm) HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for several Al% = = = =

min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 20 ppm The color of the solution from the Sample solution is not darker than that of the solution made from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution made from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution made from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (15 in 100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum zirconium pentachlorohydrate.
Aluminum Zirconium Pentachlorohydrate Solution

(Comment on this Monograph)id=m2361=Aluminum Zirconium Pentachlorohydrate Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Pentachlorohydrate Solution is a polymeric, loosely hydrated complex of basic aluminum zirconium chloride that encompasses a range of aluminum-tozirconium atomic ratios between 6.0:1 and 10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium pentachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent amount of 100 mg/mL of anhydrous aluminum zirconium pentachlorohydrate meets the requirements. B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol.
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Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Pentachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium pentachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of Aluminum by the percentage of zirconium found in the test for Content of Zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (Aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium from the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 2.1:11.51:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Sample solution: Use a weighed quantity of the Solution. Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 54 mg/mL of Aluminum Zirconium Pentachlorohydrate Solution. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 75 ppm) HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using a weighed quantity of Solution. If the solution is not clear Al% = = = =
C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium pentachlorohydrate in the Solution: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion Mr (OH),17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 500 mg of Aluminum Zirconium Pentachlorohydrate Solution to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Aluminum Zirconium Pentachlorohydrate Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and, while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.3 g of Aluminum Zirconium Pentachlorohydrate Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Al%
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after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm The color of the solution from the Sample solution is not darker than that of the solution made from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution made from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution made from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS pH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Solution to state the solvent used and the claimed concentration of anhydrous aluminum zirconium pentachlorohydrate.

ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium pentachlorohydrate in the Aluminum Zirconium Pentachlorohydrex Gly: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio from the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Pentachlorohydrex Gly to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Pentachlorohydrex Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Pentachlorohydrex Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat the solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Al%
Aluminum Zirconium Pentachlorohydrex Gly

(Comment on this Monograph)id=m2362=Aluminum Zirconium Pentachlorohydrex Gly=A-Monos.pdf) DEFINITION Aluminum Zirconium Pentachlorohydrex Gly is a derivative of Aluminum Zirconium Pentachlorohydrate in which some of the water molecules have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium atomic ratios between 6.0:1 and 10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum zirconium pentachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100 mg/mL solution meets the requirements. B. PROCEDURE Sample solution: 25 mg/mL Analysis: Heat to boiling on a hot plate, and add 60 mg of ninhydrin to 20 mL of Sample solution. Acceptance criteria: A deep violet color develops immediately.
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the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the modifications described for the Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 20 ppm The color of the solution from the Sample solution is not darker than that of the solution made from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution made from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution made from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (15 in 100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the form of glycine used and the claimed content of anhydrous aluminum zirconium pentachlorohydrate.
Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Pentachlorohydrex Gly: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of Aluminum Zirconium Pentachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 2.1:11.51:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Zirconium Pentachlorohydrex Gly. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 150 ppm) HEAVY METALS, Method I 231 [NOTEProceed as directed in the chapter, except use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for several min at 6080, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with Al% = = = =
Aluminum Zirconium Pentachlorohydrex Gly Solution

(Comment on this Monograph)id=m2363=Aluminum Zirconium Pentachlorohydrex Gly Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Pentachlorohydrex Gly Solution is a solution of Aluminum Zirconium Pentachlorohydrate in which some of the waters of hydration have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium ratios between 6.0:1 and 10.0:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium pentachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution equivalent to 100 mg/mL meets the requirements. B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol.
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C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. D. GLYCINE Sample solution: 1 g of Solution in a 50-mL beaker Add 20 mL of water, and swirl to dissolve. Analysis: Heat to boiling on a hot plate, and add 60 mg of ninhydrin for 20 mL of Sample solution. Acceptance criteria: A deep violet color develops immediately. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium pentachlorohydrate in the Solution taken: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio from the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 500 mg of Aluminum Zirconium Pentachlorohydrate Gly Solution to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Aluminum Zirconium Pentachlorohydratex Gly Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.30 g of Aluminum Zirconium Pentachlorohydrate Gly Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat the solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL Al%

of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Pentachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of Aluminum Zirconium Pentachlorohydrate taken (g) [NOTEUse the result to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 6.0:1 and 10.0:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium from the test for Content of zirconium = atomic weight of zirconium, corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride from the test for Content of chloride = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 2.1:11.51:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Sample solution: Use a weighed quantity of the Solution Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 54 mg/mL of Aluminum Zirconium Pentachlorohydrate Gly Solution. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Al% = = = =
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ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium tetrachlorohydrate in the Aluminum Zirconium Tetrachlorohydrate: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/ Ar3y) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio from the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Tetrachlorohydrate to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Tetrachlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 6 to 8 min. Add 30 40 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Tetrachlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool. Add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Tetrachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W Me = molarity of the edetate disodium VS Al%
Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 75 ppm) HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using a weighed quantity of Solution. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm The color of the solution from the Sample solution is not darker than that of the solution made from the Standard solution, and the color of the solution made from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution made from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Solution to state the solvent and form of glycine used and the claimed concentration of anhydrous aluminum zirconium pentachlorohydrate.
Aluminum Zirconium Tetrachlorohydrate

(Comment on this Monograph)id=m2365=Aluminum Zirconium Tetrachlorohydrate=A-Monos.pdf) AlyZr(OH)3y+4-xClx nH2O DEFINITION Aluminum Zirconium Tetrachlorohydrate is a polymeric, loosely hydrated complex of basic aluminum zirconium chloride that encompasses a range of aluminum-to-zirconium atomic ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum zirconium tetrachlorohydrate. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of 100 mg/mL of anhydrous aluminum zirconium tetrachloriohydrate meets the requirements.
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= volume of zinc sulfate VS consumed (mL) = molarity of the zinc sulfate VS = equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium tetrachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 2.0:1 and 5.99:1 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum from the test for Content of aluminum Ar1 = atomic weight of aluminum, 26.98 Zr% = percentage of zirconium from the test fof Content of zirconium Ar2 = atomic weight of zirconium corrected for 2% hafnium content, 92.97 Cl% = percentage of chloride from the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 1.5:10.9:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Zirconium Tetrachlorohydrate. Transfer 5.0 mL of this solution to a 50mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 150 ppm) HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for the Sample solution, if necessary. Al% z Mz Ze

Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 20 ppm The color of the solution from the Sample solution is not darker than that of the solution made from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution made from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution made from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.0 5.0, in a solution (15 in 100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum zirconium tetrachlorohydrate.
Aluminum Zirconium Tetrachlorohydrate Solution

(Comment on this Monograph)id=m2366=Aluminum Zirconium Tetrachlorohydrate Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Tetrachlorohydrate Solution is a polymeric, loosely hydrated complex of basic aluminum zirconium chloride that encompasses a range of aluminum-tozirconium atomic ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium tetrachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of 100 mg/mL of anhydrous aluminum zirconium tetrachlorohydrate meets the requirements. B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol.
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Ze
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= equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of Aluminum Zirconium Tetrachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 2.0:1 and 5.99:1 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Result = [(Al%/Ar1) + (Zr%/Ar2)] / (Cl%/Ar3) = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium from the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride from the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 1.5:10.9:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Sample solution: Use a weighed quantity of the solution. Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 54 mg/mL of Aluminum Zirconium Tetrachlorohydrate Solution. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis: To each of the beakers containing the Standard solution and the Sample solution add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. (NMT 75 ppm). HEAVY METALS, Method I 231 [NOTEProceed as directed in the chapter, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using a weighed quantity of solution. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid Al%
ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium tetrachlorohydrate in the Solution taken: Result = Al%({Ar1y + Ar2 + Mr[3y + 4(y + 1)/z] + Ar3(y + 1)/z}/ Ar1y) Al% = percentage of aluminum from the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio from the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio Ar3 = atomic weight of chlorine (Cl), 35.453 Acceptance criteria: 90.0%110.0%
OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 500 mg of Aluminum Zirconium Tetrachlorohydrate Solution to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Aluminum Zirconium Tetrachlorohydrate Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat the solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.30 g of Aluminum Zirconium Tetrachlorohydrate Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Tetrachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W Me z Mz = molarity of edetate disodium VS = volume of zinc sulfate VS consumed (mL) = molarity of zinc sulfate VS
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before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for the Sample solution, if necessary. Analysis: To each of the three tubes containing the Standard solution, the Sample solution, and the Monitor solution, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm The color of the solution from the Sample solution is not darker than that of the solution made from the Standard solution, and the color of the solution made from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution made from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Solution to state the solvent used and the claimed concentration of anhydrous aluminum zirconium tetrachlorohydrate.

ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium tetrachlorohydrate in the Aluminum Zirconium Tetrachlorohydrex Gly: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio found in the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Tetrachlorohydrex Gly to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Tetrachlorohydrex Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio).] CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Tetrachlorohydrex Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Al%
Aluminum Zirconium Tetrachlorohydrex Gly

(Comment on this Monograph)id=m2368=Aluminum Zirconium Tetrachlorohydrex Gly=A-Monos.pdf) DEFINITION Aluminum Zirconium Tetrachlorohydrex Gly is a derivative of Aluminum Zirconium Tetrachlorohydrate in which some of the water molecules have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium atomic ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum zirconium tetrachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100 mg/mL solution meets the requirements of the test for Chloride. B. PROCEDURE Sample solution: 25 mg/mL in water Analysis: Heat 20 ml of the Sample solution to boiling on a hot plate, and add 60 mg of ninhydrin. Acceptance criteria: A deep violet color develops immediately.
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several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the modifications described for the Sample solution, if necessary. Analysis Samples: Sample solution and Monitor solution To each of the three tubes containing the samples, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 20 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (15 in 100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the form of glycine used and the claimed content of anhydrous aluminum zirconium tetrachlorohydrate.
Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Tetrachlorohydrex Gly: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium tetrachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio).] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 2.0:1 and 5.99:1 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 1.5:10.9:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard iron solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Zirconium Tetrachlorohydrex Gly in water. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis Samples: Standard solution and Sample solution To each of the Samples, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: NMT 150 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for Al% = = = =
Aluminum Zirconium Tetrachlorohydrex Gly Solution

(Comment on this Monograph)id=m2369=Aluminum Zirconium Tetrachlorohydrex Gly Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Tetrachlorohydrex Gly Solution is a solution of Aluminum Zirconium Tetrachlorohydrate in which some of the waters of hydration have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 1.5:1 and 0.9:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium tetrachlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution equivalent to 100 mg/mL of anhydrous aluminum zirconium tetrachlorohydrate meets the requirements of the test for Chloride. B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a
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silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. D. GLYCINE Sample solution: 50 mg/mL in isopropyl alcohol, and filter Analysis: Heat to boiling on a hot plate, and add 60 mg of ninhydrin for 20 mL of Sample solution. Acceptance criteria: A deep violet color develops immediately. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium tetrachlorohydrate in the Solution taken: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio found in the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 500 mg of Solution to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr).[NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.3 g of Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL Al%

of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the aluminum zirconium tetrachlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar2), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar2 is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium tetrachlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum: the ratio is between 2.0:1 and 5.99:1. (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum Ar1 = atomic weight of aluminum, 26.98 Zr% = percentage of zirconium as determined in the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 1.5:10.9:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: Prepare the Sample solution using a weighed quantity of the Solution. Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 54 mg/mL of Solution in water. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis Samples: Standard solution and Sample solution To each of the two samples, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Al% Me z Mz Ze = = = =
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ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium trichlorohydrate in the Aluminum Zirconium Trichlorohydrate: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio found in the test for Aluminum/airconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Trichlorohydrate to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Trichlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Trichlorohydrate to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Trichlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W Me = molarity of the edetate disodium VS Al%
Acceptance criteria: NMT 75 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using a weighed quantity of Solution. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution if necessary. Analysis Samples: Sample solution and Monitor solution To each of the three samples, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm. The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Solution to state the solvent and form of glycine used and the claimed concentration of anhydrous aluminum zirconium tetrachlorohydrate.
Aluminum Zirconium Trichlorohydrate

(Comment on this Monograph)id=m2370=Aluminum Zirconium Trichlorohydrate=A-Monos.pdf) AlyZr(OH)3y+4-xClx nH2O DEFINITION Aluminum Zirconium Trichlorohydrate is a polymeric, loosely hydrated complex of basic aluminum zirconium chloride that encompasses a range of aluminum-to-zirconium atomic ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum zirconium trichlorohydrate. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of 100 mg/mL of anhydrous aluminum zirconium trichlorohydrate meets the requirements of the test for Chloride.
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= volume of zinc sulfate VS consumed (mL) = molarity of the zinc sulfate VS = equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar is the atomic weight of zirconium corrected for 2% hafnium content, 92.7 W = quantity of Aluminum Zirconium Trichlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 2.0:1 and 5.99:1 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Result = [(Al%/Ar1) + (Zr%/Ar2)] /(Cl%/Ar3) Al% = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium Ar2 = atomic weight of zirconium corrected for 2% hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 2.1:11.51:1 z Mz Ze

Monitor solution: Prepare as directed, using the same modifications described for the Sample solution, if necessary. Analysis Samples: Standard solution and Monitor solution To each of the three tubes containing the Samples, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 20 ppm. The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (15 in 100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the content of anhydrous aluminum zirconium trichlorohydrate.
Aluminum Zirconium Trichlorohydrate Solution

(Comment on this Monograph)id=m2371=Aluminum Zirconium Trichlorohydrate Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Trichlorohydrate Solution is a polymeric, loosely hydrated complex of basic aluminum zirconium chloride that encompasses a range of aluminum-to-zirconium atomic ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium trichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of about 100 mg/mL of anhydrous aluminum zirconium trichlorohydrate meets the requirements of the test for Chloride. B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a
IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Zirconium Trichlorohydrate in water. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis Samples: Standard soultion and Sample solution To each of the beakers containing the Samples, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: NMT 150 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide.
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Ze
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= equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium trichlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria 2.0:1 and 5.99:1 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride = atomic weight of chlorine, 35.453 Ar3 Acceptance criteria: 21:11.51:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: Prepare the Sample solution using a quantity of the Solution. Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard iron solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 54 mg/mL of Solution in water. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis Samples: Standard solution and Sample solution To each of the Samples, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: NMT 75 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using a weighed quantity of Solution. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Al%
silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium trichlorohydrate in the Solution taken: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) Al% = percentage of aluminum found in the test for Content of aluminum Ar1 = atomic weight of aluminum, 26.98 y = aluminum/zirconium atomic ratio found in the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0%
OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 500 mg of Solution to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.3 g of Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the aluminum zirconium trichlorohydrate: Result = 2.698[15.0 Me (zMz + Ze)]/W Me z Mz = molarity of the edetate disodium VS = volume of zinc sulfate VS consumed (mL) = molarity of the zinc sulfate VS
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Monitor solution: Prepare as directed, using the same modifications described for the Sample solution, if necessary. Analysis Samples: Standard solution, Sample solution, and Monitor solution To each of the three Samples, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm. The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Solution to state the solvent used and the claimed concentration of anhydrous aluminum zirconium trichlorohydrate.

ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium trichlorohydrate in the Aluminum Zirconium Trichlorohydrate Gly: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio found in the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio = atomic weight of chlorine (Cl), 35.453 Ar3 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 250 mg of Aluminum Zirconium Trichlorohydrate Gly to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 250 mg of Aluminum Zirconium Trichlorohydrate Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.15 g of Aluminum Zirconium Trichlorohydrate Gly to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Al%
Aluminum Zirconium Trichlorohydrex Gly

(Comment on this Monograph)id=m2373=Aluminum Zirconium Trichlorohydrex Gly=A-Monos.pdf) DEFINITION Aluminum Zirconium Trichlorohydrex Gly is a derivative of Aluminum Zirconium Trichlorohydrate in which some of the water molecules have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium atomic ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous aluminum zirconium trichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100 mg/mL solution meets the requirements of the test for Chloride. B. PROCEDURE Sample solution: 25 mg/mL in water Analysis: Heat to boiling on a hot plate, and add 60 mg of ninhydrin to 20 mL of Sample solution. Acceptance criteria: A deep violet color develops immediately.
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min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution, if necessary. Analysis Samples: Standard solution, Sample solution, and Monitor solution To each of the three Samples, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 20 ppm. The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (15 in 100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The label states the form of glycine used and the claimed content of anhydrous aluminum zirconium trichlorohydrate.
Calculate the percentage of aluminum (Al) in the Aluminum Zirconium Trichlorohydrate Gly: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar), where Zr% equals the percentage of zirconium as determined in the test for Content of zirconium, and Ar equals the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium trichlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 2.0:1 and 5.99:1 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 2.1:11.51:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 27 mg/mL of Aluminum Zirconium Trichlorohydrate Gly in water. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis Samples: Standard solution and Sample solution To each of the Samples, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Acceptance criteria: NMT 150 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. HEAVY METALS, Method I 231 [NOTEProceed as directed, except use the following modifications.] Sample solution: Prepare as directed in the chapter. If the solution is not clear after dilution to 40 mL, heat for several Al% = = = =
Aluminum Zirconium Trichlorohydrex Gly Solution

(Comment on this Monograph)id=m2374=Aluminum Zirconium Trichlorohydrex Gly Solution=A-Monos.pdf) DEFINITION Aluminum Zirconium Trichlorohydrex Gly Solution is a solution of Aluminum Zirconium Trichlorohydrate in which some of the waters of hydration have been displaced by glycine, calcium glycinate, magnesium glycinate, potassium glycinate, sodium glycinate, or zinc glycinate. It encompasses a range of aluminum-to-zirconium ratios between 2.0:1 and 5.99:1, and a range of (aluminum plus zirconium)-to-chloride atomic ratios between 2.1:1 and 1.51:1. The following solvents may be used: water, propylene glycol, or dipropylene glycol. It contains the equivalent NLT 90.0% and NMT 110.0% of the labeled concentration of anhydrous aluminum zirconium trichlorohydrate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution containing the equivalent of 100 mg/mL of anhydrous aluminum zirconium trichlorohydrate meets the requirements of the test for Chloride. B. PROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a
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silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of propylene glycol. C. DIPROPYLENE GLYCOL (where stated on the label) Sample solution: 200 mg/mL in isopropyl alcohol, and filter Analysis: Evaporate the filtrate to 1 mL on a steam bath: the IR absorption spectrum of a film of this solution on a silver chloride disk exhibits maxima only at the same wavelengths as that of a similar preparation of a film of dipropylene glycol. D. GLYCINE Sample solution: 50 mg/mL in isopropyl alcohol, and filter Analysis: Heat to boiling on a hot plate, and add 60 mg of ninhydrin for 20 mL of Sample solution Acceptance criteria: A deep violet color develops immediately. ASSAY PROCEDURE Analysis: Calculate the percentage of anhydrous aluminum zirconium trichlorohydrate in the Solution taken: Result = Al%({Ar1y + Ar2 + Mr[3y + 4 (y + 1)/z] + Ar3(y + 1)/z}/Ar1y) = percentage of aluminum found in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 y = aluminum/zirconium atomic ratio found in the test for Aluminum/zirconium atomic ratio = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 = molecular weight of the hydroxide anion (OH), Mr 17.01 z = (aluminum plus zirconium)/chloride atomic ratio found in the test for (Aluminum plus zirconium)/chloride atomic ratio Ar3 = atomic weight of chlorine (Cl), 35.453 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 500 mg of Solution to a 250-mL beaker Analysis: Add 100120 mL of water and 20 mL of diluted nitric acid, and swirl to dissolve. Titrate with 0.05 N silver nitrate VS using a calomel electrode and a silver billet electrode system. Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of chloride (Cl). [NOTEUse the result obtained to calculate the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ZIRCONIUM Sample: 500 mg of Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 68 min. Add 3040 mL of water and 5 mL of hydrochloric acid, and heat to boiling. Add 1 drop of xylenol orange TS, and, while still hot, titrate with 0.1 M edetate disodium VS until the color of the solution changes from pink to yellow. Perform a blank determination, and make any necessary correction. Each mL of 0.1 M edetate disodium is equivalent to 9.297 mg of zirconium (Zr). [NOTEUse the result obtained to calculate the Aluminum/zirconium atomic ratio and the (Aluminum plus zirconium)/chloride atomic ratio.] CONTENT OF ALUMINUM Sample: 0.3 g of Solution to a 150-mL beaker Analysis: Add 5 mL of water and 15 mL of hydrochloric acid. Heat this solution to boiling, and continue boiling for 5 min. Add 40 mL of water and 15.0 mL of 0.1 M edetate disodium VS. Heat the solution to boiling, and continue boiling for 5 min. Allow the solution to cool, add 1015 mL Al%

of acetic acidammonium acetate buffer TS, and adjust with ammonium hydroxide to a pH of 4.5 0.1. Add 20 mL of alcohol, and adjust with ammonium hydroxide to a pH of 4.6 0.1. Add 510 drops of dithizone TS, and titrate with 0.1 M zinc sulfate VS until the first permanent purple-pink color appears. Perform a blank determination, and make any necessary correction. Calculate the percentage of aluminum (Al) in the Solution: Result = 2.698[15.0 Me (zMz + Ze)]/W molarity of the edetate disodium VS volume of zinc sulfate VS consumed (mL) molarity of the zinc sulfate VS equivalent volume of edetate disodium VS consumed (mL) by the zirconium moiety, calculated as (Zr%/Me) (W/Ar), where Zr% is the percentage of zirconium as determined in the test for Content of zirconium, and Ar is the atomic weight of zirconium corrected for 2% hafnium content, 92.97 W = quantity of aluminum zirconium trichlorohydrate taken (g) [NOTEUse the result obtained to calculate the Aluminum/ zirconium atomic ratio and the (Aluminum plus zirconium)/ chloride atomic ratio.] ALUMINUM/ZIRCONIUM ATOMIC RATIO: Divide the percentage of aluminum found in the test for Content of aluminum by the percentage of zirconium found in the test for Content of zirconium, and multiply by 92.97/26.98, in which 92.97 is the atomic weight of zirconium corrected for 2% hafnium content, and 26.98 is the atomic weight of aluminum. Acceptance criteria: 2.0:1 and 5.99:1 (ALUMINUM PLUS ZIRCONIUM)/CHLORIDE ATOMIC RATIO Calculate the (aluminum plus zirconium)/chloride atomic ratio: Me z Mz Ze Result = [(Al%/Ar1) + (Zr%/Ar2)]/(Cl%/Ar3) = percentage of aluminum as determined in the test for Content of aluminum = atomic weight of aluminum, 26.98 Ar1 Zr% = percentage of zirconium as determined in the test for Content of zirconium = atomic weight of zirconium corrected for 2% Ar2 hafnium content, 92.97 Cl% = percentage of chloride as determined in the test for Content of chloride Ar3 = atomic weight of chlorine, 35.453 Acceptance criteria: 2.1:11.51:1 IMPURITIES Inorganic Impurities ARSENIC, Method I 211: Prepare the Sample solution using a weighed quantity of the Solution. Acceptance criteria: NMT 2 ppm LIMIT OF IRON Standard solution: Transfer 2.0 mL of Standard iron solution, prepared as directed under Iron 241, to a 50-mL beaker. Sample solution: 54 mg/mL of Solution in water. Transfer 5.0 mL of this solution to a 50-mL beaker. Analysis Samples: Standard solution and Sample solution To each of the Samples, add 5 mL of 6 N nitric acid, cover with a watch glass, and boil on a hot plate for 35 min. Allow to cool. Add 5 mL of Ammonium Thiocyanate Solution, prepared as directed under Iron 241, transfer to separate 50-mL color comparison tubes, and dilute with water to volume. Al% = = = =
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IDENTIFICATION INFRARED ABSORPTION 197S Cell: 1 mm Sample solution: 50 mg in 10 mL of 0.1 N hydrochloric acid, and filter. Transfer the filtrate to a suitable separator, add 1 mL of 5 N sodium hydroxide, and extract with 5 mL of methylene chloride. ASSAY PROCEDURE Sample: Dissolve 120 mg of Amantadine Hydrochloride in a mixture of 30 mL glacial acetic acid and 10 mL of mercuric acetate TS. Analysis: Titrate with 0.1 N perchloric acid VS, using suitable electrodes. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 18.77 mg of C10H17N HCl. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities HEAVY METALS, Method I 231: NMT 10 ppm [NOTEUse 1 mL of 1 N acetic acid in the Sample solution.] Organic Impurities PROCEDURE Internal standard solution: 50 mg/mL of adamantane in dichloromethane Sample solution: 1.0 g of Amantadine Hydrochloride in a separator. Add 20 mL of 5.0 N sodium hydroxide and 18 mL of dichloromethane, and shake for 10 min. Remove the water layer, dry the organic layer by swirling with anhydrous sodium sulfate, and allow to stand for a few min to ensure that all remaining water has been removed. Filter, collect the filtrate in a 20-mL volumetric flask, add 0.2 mL of Internal standard solution, and dilute with dichloromethane to volume. Standard solution: 10 mg of USP Amantadine Hydrochloride RS in a separator. Add 20 mL of 5.0 N sodium hydroxide and 18 mL of dichloromethane, and shake for 10 min. Remove the water layer, dry the organic layer by swirling with anhydrous sodium sulfate, and allow to stand for a few min to ensure that all remaining water has been removed. Filter, collect the filtrate in a 20-mL volumetric flask, add 0.2 mL of Internal standard solution, and dilute with dichloromethane to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.53-mm 30-m fused-silica column coated with 1.0-m G27 stationary phase Temperature: See Temperature Program Table. Increase column temperature linearly at a rate of 10/min.
Temperature Program Table Time (min) Injection port Detector Column 0 5 23 40 Temperature 220 300 70 70 250 250
Acceptance criteria: NMT 75 ppm The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution. HEAVY METALS, Method I 231 [NOTEProceed as directed in the chapter, except use the following modifications.] Sample solution: Prepare as directed in the chapter, using a quantity of Solution. If the solution is not clear after dilution to 40 mL, heat for several min between 60 and 80, and cool to room temperature. If the solution remains cloudy, repeat from the beginning with the following modification: add 3 mL of hydrochloric acid before adjustment of the pH with 6 N ammonium hydroxide. Monitor solution: Prepare as directed, using the same modifications described for Sample solution, if necessary. Analysis Samples: Standard solution, Sample soution, and Monitor solution To each of the three Samples, add 2 mL of pH 3.5 Acetate Buffer, and heat gently to between 60 and 80. To the Standard solution, add 6 drops of sodium sulfide TS. To the Sample solution and the Monitor solution, add 12 drops of sodium sulfide TS. Cool to room temperature, and dilute the contents of each tube with water to 50 mL. Gently mix each tube by inverting twice. Allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: NMT 10 ppm. The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution, and the color of the solution from the Monitor solution is the same as or darker than the color of the solution from the Standard solution. If the color of the solution from the Monitor solution is lighter than the color of the solution from the Standard solution, repeat the analysis with the following modification: after the heating step, to the Monitor solution and the Sample solution add 1.0 mL, instead of 12 drops, of sodium sulfide TS. SPECIFIC TESTS PH 791: 3.05.0, in a solution (3 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label Solution to state the solvent and form of glycine used and the claimed concentration of anhydrous aluminum zirconium trichlorohydrate.
Amantadine Hydrochloride
(Comment on this Monograph)id=m2380=Amantadine Hydrochloride=A-Monos.pdf)
C10H17N HCl Tricyclo[3.3.1.1.3,7]decan-1-amine, hydrochloride; 1-Adamantanamine hydrochloride [665-66-7].
187.71
DEFINITION Amantadine Hydrochloride contains NLT 98.5% and NMT 101.5% of C10H17N HCl.
USP 32
Carrier gas: Helium Flow rate: 4 mL/min Injection size: 2 L Injection type: Split flow is 200 mL/min with a split flow ratio of 50:1. System suitability Sample: Standard solution [NOTEThe relative retention times for adamantane and amantadine are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 20 between adamantane and amantadine Relative standard deviation: NMT 5.0% determined from the peak response ratios of amantadine to adamantane Analysis Samples: Sample solution and Standard solution Calculate the percentage of each impurity in the portion of C10H17N HCl taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of each impurity to adamantane from the Sample solution = peak response ratio of amantadine to RS adamantane from the Standard solution = concentration of USP Amantadine Hydrochloride CS RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria Individual impurities: NMT 0.3% Total impurities: NMT 1.0% RU SPECIFIC TESTS PH 791: 3.05.5, in a solution (1 in 5) CLARITY AND COLOR OF SOLUTION Sample: Dissolve 2 g in 10 mL of water. Acceptance criteria: Solution is clear and nearly colorless. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amantadine Hydrochloride RS
Official Monographs / Amantadine 169

Standard stock solution: 2 mg/mL of USP Amantadine Hydrochloride RS in water Standard solution: Pipet 25.0 mL of Standard stock solution into a 250-mL separator, and add 25 mL of 2.0 N sodium hydroxide and 50.0 mL of Internal standard solution. Shake for 60 min, and collect the hexane layer (Standard solution). Sample solution: Transfer NLT 20 Capsules to a 200-mL volumetric flask. Add 40 mL of 0.1 N hydrochloric acid, and heat gently to achieve complete dissolution. Cool, and dilute with water to volume. Pipet 5.0 mL of the solution into a 250-mL separator, and add 40 mL of 1.0 N sodium hydroxide and 50.0 mL of Internal standard solution. Shake for 60 min, and collect the hexane layer (Sample solution). Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.22-m; glass column packed with 10% phase G1 on 100- to 120-mesh support S1A Temperature Column: 115 Injection port: 250 Detector block: 250 Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2 between napthalene and amantadine Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage C10H17N HCl taken: Result = (RU/RS) (CS/CU) 100 = peak response ratios from the Sample solution = peak response ratios from the Standard solution = concentration of USP Amantadine Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 95.0%105.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Internal standard solution: 0.054 mg/mL of naphthalene in hexane Standard stock solution: 0.1 mg/mL of USP Amantadine Hydrochloride RS in water Standard solution: Pipet 15.0 mL of Standard stock solution into a 50-mL screw-capped test tube, add 5.0 mL of 5 N sodium hydroxide and 10.0 mL of Internal standard solution, and shake for 60 min. Collect the hexane layer. Sample solution: Filter 15.0 mL of the solution under test, and place into a 50-mL screw-capped test tube. Pipet 5.0 mL of 5 N sodium hydroxide and 10.0 mL of the Internal standard solution into the test tube, and shake for 60 min. Collect the hexane layer (Sample solution). Chromatographic system: Proceed as directed in the Assay. Injection size: 2.5 L Analysis Sample: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C10H17N HCl is dissolved in 45 min. RU RS CS
Amantadine Hydrochloride Capsules

(Comment on this Monograph)id=m2390=Amantadine Hydrochloride Capsules=A-Monos.pdf) DEFINITION Amantadine Hydrochloride Capsules contain NLT 95.0% and NMT 105.0% of the labeled amount of amantadine hydrochloride (C10H17N HCl). IDENTIFICATION INFRARED ABSORPTION 197S Cell: 1 mm Sample solution: Place the contents of Capsules, equivalent to 200 mg of amantadine hydrochloride, in a vessel, dissolve in 0.1 N hydrochloric acid, and filter. Transfer the filtrate to a separator, add 1 mL of 5 N sodium hydroxide, and extract with 5 mL of methylene chloride. Filter the extract through anhydrous sodium sulfate, and rinse the anhydrous sodium sulfate with 2 mL of methylene chloride. ASSAY PROCEDURE Internal standard solution: hexane
0.4 mg/mL of naphthalene in
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Meet the requirements RS CS
USP 32
= peak response ratios from the Standard solution = concentration of USP Amantadine Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of amantadine hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amantadine Hydrochloride RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amantadine Hydrochloride RS
Amantadine Hydrochloride Oral Solution

(Comment on this Monograph)id=m2420=Amantadine Hydrochloride Oral Solution=A-Monos.pdf) DEFINITION Amantadine Hydrochloride Oral Solution contains NLT 95.0% and NMT 105.0% of the labeled amount of amantadine hydrochloride (C10H17N HCl). IDENTIFICATION INFRARED ABSORPTION 197S Cell: 1 mm Sample solution: Place a volume of Oral Solution, equivalent to about 200 mg of amantadine hydrochloride, in a vessel, dissolve in 0.1 N hydrochloric acid, and filter. Transfer the filtrate to a separator, add 10 mL of 0.5 N sodium hydroxide, and extract with 5 mL of methylene chloride. Filter the extract through anhydrous sodium sulfate, and rinse the anhydrous sodium sulfate with 2 mL of methylene chloride. ASSAY PROCEDURE Internal standard solution: 0.4 mg/mL of naphthalene in hexane Standard stock solution: 2 mg/mL of USP Amantadine Hydrochloride RS in water Standard solution: Pipet 25.0 mL of Standard stock solution into a 250-mL separator, add 25 mL of 2.0 N sodium hydroxide and 50.0 mL of Internal standard solution. Shake for 60 min, and collect the hexane layer. Sample solution: Pipet 5.0 mL of the Oral Solution into a 250-mL conical flask, and add 45 mL of 1.0 N sodium hydroxide and 50.0 mL of Internal standard solution. Shake for 60 min, and collect the hexane layer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.22-m glass column packed with 10% phase G1 on 100- to 120-mesh support S1A Temperature Column: 115 Injection port: 250 Detector block: 250 Injection size 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2 between napthalene and amantadine Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H17N HCl in the portion of Oral Solution taken: Result = (RU/RS) (CS/CU) 100 RU = peak response ratios from the Sample solution
Amcinonide
(Comment on this Monograph)id=m2550=Amcinonide=AMonos.pdf)
502.57 C28H35FO7 Pregna-1,4-diene-3,20-dione, 21-(acetyloxy)-16,17[cyclopentylidenebis(oxy)]-9-fluoro-11-hydroxy-, (11,16)-; 9-Fluoro-11,16,17,21-tetrahydroxypregna-1,4-diene-3,20dione cyclic 16,17-acetal with cyclopentanone, 21-acetate [51022-69-6]. DEFINITION Amcinonide contains NLT 97.0% and NMT 102.0% of C28H35FO7, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 238 nm Solution: 40 g/mL in methanol Acceptance criteria: Absorptivities do not differ by more than 3.0%, calculated on the dried basis ASSAY PROCEDURE Solution A: Acetonitrile and water (7:13) Solution B: Acetonitrile and water (7:3) System suitability solution: 12.5 g/mL of butylparaben and 20 g/mL of USP Amcinonide RS in Solution B Standard solution: 0.02 mg/mL of USP Amcinonide RS in Solution B Sample stock solution: 0.2 mg/mL of Amcinonide in Solution B [NOTESonicate for 5 min.] Sample solution: 0.02 mg/mL of Sample stock solution in Solution B Mobile phase: See the gradient table below.
Time (min) 0 2.5 10 Solution A (%) 100 100 0 Solution B (%) 0 0 100
Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for butylparaben and amcinonide are 0.78 and 1.0, respectively.] Suitability requirements Resolution: NLT 8.0 between butylparaben and amcinonide, System suitability solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C28H35FO7 in the portion of Amcinonide taken: Result = (rU/rS) (CS/CU) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Amcinonide RSin the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 97.0%102.0% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: rU rS CS
Official Monographs / Amcinonide 171

Application volume: 25 L Developing solvent: Ether Analysis Samples: Standard solution and Sample solution Develop the chromatogram until the solvent front has moved about 12 cm above the line of application. Acceptance criteria: The intensity and the RF value of the principal spot of the Sample solution are similar to those of the Standard solution. ASSAY PROCEDURE Solution A: Acetonitrile and water (7:13) Solution B: Acetonitrile and water (7:3) System suitability solution: 12.5 g/mL of butylparaben and 20 g/mL of USP Amcinonide RS in Solution B Standard solution: 0.02 mg/mL of USP Amcinonide RS in Solution B Sample solution: Transfer a quantity of Cream, equivalent to 10 mg of Amcinonide, to a 50-mL volumetric flask. Add 5 mL of Solution B and 15 mL of acetonitrile, and heat over a steam bath until dissolved. Add 20 mL of Solution B while hot, cool to room temperature, dilute with Solution B to volume, and refrigerate for 30 min. Vigorously shake the solution to disperse the mixture, and filter while cold. Transfer 5 mL of this solution to a 50-mL volumetric flask, and dilute with Solution B to volume. Mobile phase: See the gradient table below.
NMT 20 ppm
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +89.4 to +94.0 Sample solution: 10 mg/mL, in chloroform LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amcinonide RS
Amcinonide Cream
(Comment on this Monograph)id=m2555=Amcinonide Cream=A-Monos.pdf) DEFINITION Amcinonide Cream is Amcinonide in a suitable cream base. It contains NLT 90.0% and NMT 115.0% of the labeled amount of amcinonide (C28H35FO7). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 100 g/mL of USP Amcinonide RS in chloroform Sample solution: Place 2 g of the Cream in a 150-mL beaker, add 50 mL of chloroform and 15 g of anhydrous sodium sulfate, and stir with a glass rod to dissolve the specimen. Filter the solution, and clarify the filtrate, if necessary, by the further addition of anhydrous sodium sulfate and a second filtration. Evaporate the filtrate to dryness, and dissolve the residue in chloroform to obtain a solution containing 100 g/mL of amcinonide.
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for butylparaben and amcinonide are about 0.78 and 1.0, respectively.] Suitability requirements Resolution: NLT 8.0 between butylparaben and amcinonide, System suitability solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C28H35FO7 in the portion of Cream taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Amcinonide RS in the Standard solution (mg/mL) = nominal concentration of amcinonide in the Sample solution (mg/mL)
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90.0%115.0%
USP 32
quantitatively with the same solvent mixture to obtain a solution having a concentration of 0.2 mg/mL of amcinonide. Cool to room temperature, dilute with acetonitrile to volume, and filter. Sample solution: Dilute Sample stock solution with Solution B (5 in 50). Mobile phase: See the gradient table below.
PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. PH 791: 3.55.2 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amcinonide RS
Amcinonide Ointment
(Comment on this Monograph)id=m2560=Amcinonide Ointment=A-Monos.pdf) DEFINITION Amcinonide Ointment is Amcinonide in a suitable ointment base. It contains NLT 90.0% and NMT 115.0% of the labeled amount of amcinonide (C28H35FO7). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 100 g/mL of USP Amcinonide RS in chloroform Sample solution: Place 2 g of the Ointment in a 150-mL beaker, add 50 mL of chloroform and 15 g of anhydrous sodium sulfate, and stir with a glass rod to dissolve the specimen. Filter the solution, and clarify the filtrate, if necessary, by the further addition of anhydrous sodium sulfate and a second filtration. Evaporate the filtrate to dryness, and dissolve the residue in chloroform to obtain a solution containing 100 g/mL of amcinonide. Application volume: 25 L Developing solvent: Ether Analysis Samples: Standard solution and Sample solution Develop the chromatogram until the solvent front has moved about 12 cm above the line of application. Acceptance criteria: The intensity and the RF value of the principal spot of the Sample solution are similar to those of the Standard solution. ASSAY PROCEDURE Solution A: Acetonitrile and water (7:13) Solution B: Acetonitrile and water (7:3) System suitability solution: 12.5 g/mL of butylparaben and 20 g/mL of USP Amcinonide RS in Solution B Standard solution: 0.02 mg/mL of USP Amcinonide RS in Solution B Sample stock solution: Dissolve Ointment in a suitable volume of a mixture of acetonitrile and chloroform (4:1) by heating in a hot water bath, cooling, and adjusting
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for butylparaben and amcinonide are 0.78 and 1.0, respectively.] Suitability requirements Resolution: NLT 8.0 between butylparaben and amcinonide, System suitability solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C28H35FO7 in the portion of Ointment taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amcinonide RS in the Standard solution (mg/mL) = nominal concentration of amcinonide in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% rU rS CS PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amcinonide RS
USP 32
Official Monographs / Amifostine 173

Amifostine
(Comment on this Monograph)id=m2600=Amifostine=AMonos.pdf)
C5H15N2O3PS 3H2O 268.27 Ethanethiol, 2-[(3-aminopropyl)amino]-, dihydrogen phosphate (ester), trihydrate; S-[2-(3-Aminopropyl)amino]ethyl]dihydrogen phosphorothioate, trihydrate [112901-68-5]. DEFINITION Amifostine contains NLT 78.0% and NMT 82.0% of C5H15N2O3PS, calculated on the as-is basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid in 1200 mL of HPLC grade water. Prepare a mixture of this solution and acetonitrile (90:10). Standard solution: 3 mg/mL of USP Amifostine RS in water [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] Sample solution: 3 mg/mL of Amifostine in water [NOTE Inject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 30 [NOTEThe temperature of the solutions to be injected is maintained at 28.] Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard solution and Sample solution Suitability requirements Column efficiency: NLT 7500 theoretical plates, Standard solution and Sample solution Tailing factor: NMT 2, Standard solution and Sample solution Relative standard deviation: NMT 2.0%, Standard solution and Sample solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C5H15N2O3PS in the portion of Amifostine taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Amifostine RS in the Standard solution (mg/mL) = concentration of Amifostine in the Sample solution (mg/mL)
IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid in 1200 mL of HPLC grade water, add 400 L of trifluoroacetic acid, and adjust with triethylamine to a pH of 2.5. Prepare a mixture of this solution and acetonitrile (68:32). Blank: Water Standard thiol solution: 0.124 mg/mL of USP Amifostine Thiol RS in water [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] System suitability solution: 5 mg/mL of USP Amifostine RS in Standard thiol solution [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 12 h if maintained at about 5.] Sample solution: 50 mg/mL of Amifostine in water [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 30 [NOTEThe temperature of the solutions to be injected is maintained at 28.] Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard thiol solution and System suitability solution Suitability requirements Resolution: NLT 2.0 between amifostine and amifostine thiol, System suitability solution Tailing factor: NMT 4.0, System suitability solution and Standard solution Capacity factor: More than 0.5, System suitability solution and Standard solution Column efficiency: NLT 2300 theoretical plates calculated for the amifostine thiol peak, System suitability solution and Standard solution Relative standard deviation: NMT 4.0%, System suitability solution and Standard solution Analysis Samples: Blank, Standard thiol solution, and Sample solution [NOTEMeasure the responses of all the peaks, excluding the peaks corresponding to those from the Blank.] Calculate the percentage of amifostine thiol in the portion of Amifostine taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU = peak response of amifostine thiol from the Sample solution = peak response of amifostine thiol from the Standard thiol solution = concentration of amifostine thiol dihydrochloride in the Standard thiol solution (mg/mL) = concentration of Amifostine in the Sample solution (mg/mL)
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= molecular weight of amifostine thiol, 134.24 =molecular weight of amifostine thiol dihydrochloride, 207.17 Calculate the percentage of each of the other impurities in the portion of Amifostine taken: Result = (ri/rU) 100 Mr1 Mr2
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Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard solution and Sample solution Suitability requirements Column efficiency: NLT 7500 theoretical plates, Standard solution and Sample solution Tailing factor: NMT 2, Standard solution and Sample solution Relative standard deviation: NMT 2.0%, Standard solution and Sample solution Analysis Samples: Standard solution and Sample solution Calculate the percentage C5H15N2O3PS in the portion of Amifostine for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Amifostine RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: requirements Meets the rU rS CS
= peak response of each impurity from the Sample solution = peak response of amifostine from the Sample rU solution Acceptance criteria Any individual impurity, excluding amifostine thiol: NMT 0.1% Total impurities including amifostine thiol: NMT 0.3% ri SPECIFIC TESTS X-RAY DIFFRACTION 941: Its X-ray diffraction pattern conforms to that of USP Amifostine RS, similarly determined. PH 791: 6.57.5, in a solution (5 in 100) WATER DETERMINATION, Method Ic 921 Sample solution: To 100.0 mg of Amifostine, contained in a stoppered centrifuge tube, add 10.0 mL of the solution of N-ethylmaleimide in methanol (4 in 100), and sonicate for 15 min. Shake to disperse, and sonicate for an additional 15 min. Use 1.0 mL of the supernatant. Acceptance criteria: 19.2%21.2% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store in a refrigerator. USP REFERENCE STANDARDS 11 USP Amifostine RS USP Amifostine Thiol RS
Amifostine for Injection

(Comment on this Monograph)id=m2603=Amifostine for Injection=A-Monos.pdf) DEFINITION Amifostine for Injection is a sterile, crystalline substance suitable for parenteral use. It contains NLT 90.0% and NMT 110.0% of the labeled amount of amifostine (C5H15N2O3PS). ASSAY PROCEDURE Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid in 1200 mL of HPLC grade water. Prepare a degassed mixture of this solution and acetonitrile (90:10). Standard solution: 3 mg/mL of USP Amifostine RS [NOTE Inject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] Sample stock solution: Equivalent to 10 mg/mL of amifostine, from Amifostine for Injection Sample solution: Transfer 6.0 mL of Sample stock solution to a 25-mL volumetric flask, add 6.5 mL of water, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 30 [NOTEThe temperature of the solutions to be injected is maintained at 28.]
IMPURITIES Organic Impurities PROCEDURE Mobile phase: 1.0 mL of nonafluorobutane sulfonic acid in 1200 mL of HPLC grade water, add 400 L of trifluoroacetic acid, and adjust with triethylamine to a pH of 2.5. Prepare a degassed mixture of this solution and acetonitrile (68:32). Blank: Water Standard disulfide solution: 0.186 mg/mL of USP Amifostine Disulfide RS [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] Standard thiol solution: 0.802 mg/mL of USP Amifostine Thiol RS [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] System suitability solution: 5 mg/mL of USP Amifostine RS in Standard thiol solution [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 12 h if maintained at about 5.] Sample solution: 50 mg/mL of amifostine, from Amifostine for Injection [NOTEInject immediately after preparation, or refrigerate until use. The solution is stable for 48 h if maintained at about 5.] Chromatographic system (See Chromatography 621, System Suitability.) Mode LC Detector: UV, 220 nm and 247 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 30 [NOTEThe temperature of the solutions to be injected is maintained at 28.]
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Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard disulfide solution, Standard thiol solution, and System suitability solution Suitability requirements Capacity factor, k: More than 0.5 for amifostine thiol; more than 2.2 for amifostine disulfide, Standard disulfide solution, Standard thiol solution, and System suitability solution Column efficiency: NLT 2300 theoretical plates, for amifostine thiol; not more than 2000 for amifostine, for amifostine disulfide Tailing factor: NMT 4.0 for amifostine thiol; NMT 4.5 for amifostine disulfide, Standard disulfide solution, Standard thiol solution, and System suitability solution Relative standard deviation: NMT 4.0% for amifostine thiol; NMT 4.0% for amifostine disulfide, System suitability solution, Standard disulfide solution, andStandard thiol solution Analysis Samples: Blank, Standard disulfide solution, Standard thiol solution and Sample solution [NOTEMeasure the responses of all the peaks, excluding the peaks corresponding to those from the Blank.] Calculate the percentage of amifostine thiol in the portion of Amifostine for Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = amifostine thiol peak responses at 220 nm, from the Sample solution rS = amifostine thiol peak responses at 220 nm, from the Standard thiol solution = concentration of amifostine thiol CS dihydrochloride in the Standard thiol solution (mg/mL) = concentration of the Sample solution (mg/mL) CU = molecular weight of amifostine thiol, 134.24 Mr1 = molecular weight of amifostine thiol Mr2 dihydrochloride, 207.17 Calculate the percentage of amifostine disulfide in the portion of Amifostine for Injection taken: Result = (rU/rS) (CS/CU) (Mr3/Mr4) 100 = peak responses at 247 nm, from the Sample solution = peak responses at 247 nm, from the Standard rS disulfide solution = concentration of USP Amifostine Disulfide RS in CS the Standard disulfide solution (mg/ml) = concentration for the Sample solution (mg/mL) CU = molecular weight of amifostine disulfide, 266.47 Mr3 = molecular weight of amifostine disulfide Mr4 tetrahydrochloride, 412.31 Acceptance criteria: NMT 2.0% of total impurities, including amifostine thiol and amifostine disulfide Calculate the percentage of each of the other impurities in the portion of Amifostine for Injection taken: rU Result = (ri/rA) 100 ri rA = peak response for each impurity from the Sample solution = peak response for amifostine from the Sample solution rU
Official Monographs / Amikacin 175

Acceptance criteria: NMT 0.1% of any individual impurity except amifostine thiol is found. SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. When constituted with 0.9% Sodium Chloride Injection, the solution must completely dissolve in 45 s. X-RAY DIFFRACTION 941: Its X-ray diffraction pattern conforms to that of USP Amifostine RS, similarly determined. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 6.57.5, in a solution constituted as directed in the labeling WATER DETERMINATION, Method Ic 921 Sample solution: To 100.0 mg of Amifostine for Injection, contained in a stoppered centrifuge tube, add 10.0 mL of the solution of N-ethylmaleimide in methanol (4 in 100), and sonicate for 15 min. Shake to disperse, and sonicate for an additional 15 min. Use 1.0 mL of the supernatant. Acceptance criteria: 18.0%22.0% PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections BACTERIAL ENDOTOXINS TEST 85: Contains NMT 0.2 USP Endotoxin Unit/mg of amifostine OTHER REQUIREMENTS: Meets the requirements for Labeling under Injections 1. It also meets the requirements for Identification under Amifostine. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight Containers for Sterile Solids as described under Injections 1, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Amifostine RS USP Amifostine Disulfide RS USP Amifostine Thiol RS USP Endotoxin RS
Amikacin
(Comment on this Monograph)id=m2610=Amikacin=AMonos.pdf)
C22H43N5O13 585.60 D-Streptamine, O-3-amino-3-deoxy--D-glucopyranosyl-(16)O-[6-amino-6-deoxy--D-glucopyranosyl(14)]-N1-(4amino-2-hydroxy-1-oxobutyl)-2-deoxy-, (S)-; O-3-Amino-3-deoxy--D-glucopyranosyl(14)-O-[6-amino-6deoxy--D-glucopyranosyl(16)]-N3-(4-amino-L-2hydroxybutyryl)-2-deoxy-L-streptamine [37517-28-5]. DEFINITION Amikacin has a potency of NLT 900 g of C22H43N5O13 per mg, calculated on the anhydrous basis.
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Acceptance criteria: NLT 900 g
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IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 6 mg/mL Sample solution: 6 mg/mL Solution A: Sample solution and the Standard solution (1:1) Application volume: 3 L Developing solvent system: Methanol, chloroform, and ammonium hydroxide (12:5:7) Spray reagent: 10 mg/mL of ninhydrin in a mixture of butyl alcohol and pyridine (100:1) Analysis Samples: Standard solution, Sample solution, and Solution A Proceed as directed in the chapter, except to develop the chromatogram by continuous flow for 5.5 h. Remove the plate from the chamber, allow the solvent to evaporate, and heat the plate at 110 for 15 min. Spray the plate with Spray reagent, and immediately locate the spots. Acceptance criteria: Amikacin appears as a pink spot, and the spots obtained from the Sample solution and Solution A correspond in distance from the origin to that obtained from the Standard solution. B. The retention time of the peak for amikacin of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.115 N sodium hydroxide System suitability solution: 0.02 mg/mL of USP Amikacin RS and 0.008 mg/mL of USP Kanamycin Sulfate RS Standard solution: 0.02 mg/mL of USP Amikacin RS Sample solution: 0.02 mg/mL of Amikacin Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Electrochemical detector, a gold working electrode, and a pH silversilver chloride reference electrode [NOTEThe electrochemical detector is used in the integrated amperometric mode with a range of 300 nC, an output of 1 V full scale, a rise time of 0.5 s, positive polarity, potential E = 0.04 V; t1 = 200 ms; E2 = 0.8 V; t2 = 190 ms; E3 = 0.8 V; and t3 = 190 ms.] Guard column: Packing L47 Analytical column: 4-mm 25-cm; packing L47 Flow rate: 0.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times of kanamycin and amikacin are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 between kanamycin and amikacin, System suitability solution Tailing factor: NMT 2, Standard solution Relative standard deviation: NMT 3%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C22H43N5O13 in each mg of Amikacin taken: Result = (rU/rS) (CS/CU) E rU rS CS CU E = amikacin peak areas from the Sample solution = amikacin peak areas from the Standard solution = concentration of USP Amikacin RS in the Standard solution (mg/mL) = concentration for the Sample solution (mg/mL) = designated amikacin content of USP Amikacin RS (g/mg)
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 1.0%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S Sample solution: 20 mg/mL Acceptance criteria: +97 to +105 CRYSTALLINITY 695: Meets the requirements PH 791: 9.511.5, in a solution containing 10 mg/mL WATER DETERMINATION, Method I 921: NMT 8.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amikacin RS USP Kanamycin Sulfate RS
Amikacin Sulfate
(Comment on this Monograph)id=m2630=Amikacin Sulfate=AMonos.pdf)
D-Streptamine,
C22H43N5O13 2H2SO4 781.76 O-3-amino-3-deoxy--D-glucopyranosyl-(16)O-[6-amino-6-deoxy--D-glucopyranosyl-(14)]-N1-(4amino-2-hydroxy-1-oxobutyl)-2-deoxy-, (S)-, sulfate (1:2) (salt); O-3-Amino-3-deoxy--D-glucopyranosyl-(14)-O-[6-amino-6deoxy--D-glucopyranosyl-(16)]-N3-(4-amino-L-2hydroxybutyryl)-2-deoxy-L-streptamine sulfate (1:2) [39831-55-5; 3517-28-5].
DEFINITION Amikacin Sulfate having a molar ratio of amikacin to H2SO4 of 1:2 contains the equivalent of NLT 674 g and NMT 786 g of amikacin (C22H43N5O13) per mg, calculated on the dried basis; Amikacin Sulfate having a molar ratio of amikacin to H2SO4 of 1:1.8 contains the equivalent of NLT 691 g and NMT 806 g of amikacin (C22H43N5O13) per mg, calculated on the dried basis. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 6 mg/mL in water Sample solution: 6 mg/mL in water Solution A: Sample solution and the Standard solution (1:1) Application volume: 3 L Developing solvent system: Methanol, ammonium hydroxide, and chloroform (12:7:5) Spray reagent: 10 mg/mL of ninhydrin in a mixture of butyl alcohol and pyridine (100:1) Analysis Samples: Standard solution, Sample solution, and Solution A Proceed as directed in the chapter, except to develop the chromatogram by continuous flow for 5.5 h. Remove the plate from the chamber, allow the solvent to evaporate,
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and heat the plate at 110 for 15 min. Spray the plate with Spray reagent, and immediately locate the spots. Acceptance criteria: Amikacin appears as a pink spot, and the spots of the Sample solution and Solution A correspond in distance from the origin to that of the Standard solution. B. The retention time of the peak for amikacin of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.115 N sodium hydroxide System suitability solution: 0.02 mg/mL of USP Amikacin RS and 0.008 mg/mL of USP Kanamycin Sulfate RS in water Standard solution: 0.02 mg/mL of USP Amikacin RS in water Sample solution: Equivalent to 0.02 mg/mL of amikacin, from Amikacin Sulfate, in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Electrochemical detector, a gold working electrode, and a pH silver-silver chloride reference electrode [NOTEThe electrochemical detector is used in the integrated amperometric mode with a range of 300 nC, an output of 1 V full scale, a rise time of 0.5 s, positive polarity, potential E = 0.04 V; t1 = 200 ms; E2 = 0.8 V; t2 = 190 ms; E3 = 0.8 V; and t3 = 190 ms.] Column Guard column: Packing L47 Analytical column: 4-mm 25-cm; packing L47 Flow rate: 0.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for kanamycin and amikacin are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 between kanamycin and amikacin, System suitability solution Tailing factor: NMT 2, Standard solution Relative standard deviation: NMT 3%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C22H43N5O13 in each mg of Amikacin Sulfate taken: Result = (rU/rS) (CS/CU) E = amikacin peak areas from the Sample solution = amikacin peak areas from the Standard solution = concentration of USP Amikacin RS in the Standard solution (mg/mL) = concentration for the Sample solution CU (mg/mL) E = designated amikacin content of USP Amikacin RS (g/mg) Acceptance criteria: Amikacin Sulfate having a molar ratio of amikacin to H2SO4 of 1:2 contains the equivalent of NLT 674 g and NMT 786 g of C22H43N5O13 per mg; Amikacin Sulfate having a molar ratio of amikacin to H2SO4 of 1:1.8 contains the equivalent of NLT 691 g and NMT 806 g of C22H43N5O13 per mg rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 1.0%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid
Official Monographs / Amikacin 177

SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781: +76 to +84 Sample solution: 20 mg/mL, in water CRYSTALLINITY 695: Meets the requirements PH 791: 2.04.0 (1:2 salt), or 6.07.3 (1:1.8 salt), in a solution containing 10 mg/mL LOSS ON DRYING 731: Dry 100 mg, in a vacuum at a pressure not exceeding 5 mm of mercury at 110 for 3 h: it loses NMT 13.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate whether its molar ratio of amikacin to H2SO4 is 1:2 or 1:1.8. USP REFERENCE STANDARDS 11 USP Amikacin RS USP Kanamycin Sulfate RS
Amikacin Sulfate Injection

(Comment on this Monograph)id=m2640=Amikacin Sulfate Injection=A-Monos.pdf) DEFINITION Amikacin Sulfate Injection is a sterile solution of Amikacin Sulfate in Water for Injection, or of Amikacin in Water for Injection prepared with the aid of Sulfuric Acid. It contains NLT 90.0% and NMT 120.0% of the labeled amount of amikacin (C22H43N5O13). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 6 mg/mL Sample solution: 6 mg/mL Solution A: Sample solution and the Standard solution (1:1) Application volume: 3 L Developing solvent system: Methanol, chloroform, and ammonium hydroxide (12:5:7) Spray reagent: 10 mg/mL of ninhydrin in a mixture of butyl alcohol and pyridine (100:1) Analysis Samples: Standard solution, Sample solution, and Solution A Proceed as directed in the chapter, except to develop the chromatogram by continuous flow for 5.5 h. Remove the plate from the chamber, allow the solvent to evaporate, and heat the plate at 110 for 15 min. Spray the plate with Spray reagent, and immediately locate the spots. Acceptance criteria: Amikacin appears as a pink spot, and the spots obtained from the Sample solution and the Solution A correspond in distance from the origin to that obtained from the Standard solution. B. The retention time of the peak for amikacin of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 0.115 N sodium hydroxide System suitability solution: 0.02 mg/mL of USP Amikacin RS and 0.008 mg/mL of USP Kanamycin Sulfate RS Standard solution: 0.02 mg/mL of USP Amikacin RS Sample solution: 0.02 mg/mL of amikacin, from the Injection Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: Electrochemical detector, a gold working electrode, and a pH silversilver chloride reference electrode [NOTEThe electrochemical detector is used in the integrated amperometric mode with a range of 300 nC, an output of 1 V full scale, a rise time of 0.5 s, positive polarity, potential E = 0.04 V; t1 = 200 ms; E2 = 0.8 V; t2 = 190 ms; E3 = 0.8 V; and t3 = 190 ms.] Column Guard column: Packing L47 Analytical column: 4-mm 25-cm; packing L47 Flow rate: 0.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times of kanamycin and amikacin are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 between kanamycin and amikacin, System suitability solution Tailing factor: NMT 2, Standard solution Relative standard deviation: NMT 3%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H43N5O13 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) E F100 = amikacin peak areas from the Sample solution = amikacin peak areas from the Standard solution = concentration of USP Amikacin RS in the Standard solution (mg/mL) CU = nominal concentration of amikacin in the Sample solution E = designated amikacin content of USP Amikacin RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% SPECIFIC TESTS PH 791: 3.55.5 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections BACTERIAL ENDOTOXINS TEST 85: NMT 0.33 USP Endotoxin Unit/mg of amikacin OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I or Type III glass. USP REFERENCE STANDARDS 11 USP Amikacin RS USP Endotoxin RS USP Kanamycin Sulfate RS rU rS CS
Amiloride Hydrochloride
(Comment on this Monograph)id=m2650=Amiloride Hydrochloride=A-Monos.pdf)
302.12 C6H8ClN7O HCl 2H2O Pyrazinecarboxamide, 3,5-diamino-N-(aminoiminomethyl)-6chloro-, monohydrochloride dihydrate; N-Amidino-3,5-diamino-6-chloropyrazinecarboxamide monohydrochloride dihydrate [17440-83-4]. DEFINITION Amiloride Hydrochloride contains NLT 98.0% and NMT 101.0% of C6H8ClN7O HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Sample solution: 600 g/mL of water, diluted quantitatively and stepwise with 0.1 N hydrochloric acid to a concentration of about 9.6 g/mL C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Sample: 450 mg Analysis: Dissolve in 100 mL of glacial acetic acid, add 10 mL of mercuric acetate TS and 15 mL of dioxane. Add crystal violet TS. Titrate with 0.1 N perchloric acid VS to a blue endpoint. Perform a blank determination (See Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 26.61 mg of C6H8ClN7O HCl. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Standard solutions: Prepare a series of solutions, A, B, C, D, E, and F, of USP Amiloride Hydrochloride RS in a mixture of methanol and chloroform (4:1) having concentrations of 4000, 40, 20, 8, 4, and 2 g/mL, respectively. Sample solution: 4 mg/mL of Amiloride Hydrochloride in methanol and chloroform (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chormatography.
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Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel previously washed with methanol Application volume: 5 L Developing solvent system: Tetrahydrofuran and 3 N ammonium hydroxide (15:2) Analysis Samples: Standard solutions A, B, C, D, E, and F and the Sample solution Proceed as directed under General Chapter. Dry the spots with a stream of nitrogen, and develop the chromatograms in the solvent system, until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow to air-dry, and examine the plate under long-wavelength UV light: the RF value of the principal spot of the Sample solution corresponds to that of Standard solution A. Estimate the levels of any additional spots observed in the chromatogram of the Sample solution by comparison with the principal spots in the chromatograms of Standard solutions B, C, D, E, and F. Acceptance criteria: The sum of the intensities of any additional spots observed is NMT that of the principal spot obtained from Standard solution B (equivalent to 1%). SPECIFIC TESTS ACIDITY Sample: 1.0 g Analysis: Dissolve in 100 mL of a mixture of methanol and water (1:1). Titrate with 0.10 N sodium hydroxide. Acceptance criteria: NMT 0.30 mL is required (0.1% as HCl) LOSS ON DRYING (See Thermal Analysis 891) [NOTEThe quantity taken for the determination may be adjusted, if necessary, for instrument sensitivity.] Sample: 10 mg of Amiloride Hydrochloride Analysis: Determine the percentage of volatile substances by thermogravimetric analysis on an appropriately calibrated instrument, using the Sample. Heat the specimen at the rate of 10/min between ambient temperature and 225 in an atmosphere of nitrogen at a flow rate of 40 mL/min. From the thermogram determine the accumulated loss in weight between ambient temperature and about 200 on the plateau. Acceptance criteria: It loses NLT 11.0% and NMT 13.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amiloride Hydrochloride RS
Official Monographs / Amiloride 179

Standard solution: 0.2 mg/mL of USP Amiloride Hydrochloride RS in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Developing solvent system: Tetrahydrofuran and 3 N ammonium hydroxide (22:3) Application volume: 10 L Analysis Samples: Sample solution and Standard solution Remove the plate from the developing chamber, air-dry, and examine under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, water, and Buffer solution (25:71:4) Solution A: 136 g of monobasic potassium phosphate in 800 mL of water. Adjust by the addition of phosphoric acid, with mixing, to a pH of 3.0. Dilute with water to 1000 mL. Standard stock solution: 1.0 mg/mL of amiloride hydrochloride, from USP Amiloride Hydrochloride RS in methanol Standard solution: 5.0 mL of the Standard stock solution to a 50-mL volumetric flask. Add 10 mL of methanol, 2.0 mL of 0.1 N hydrochloric acid, and dilute with water to volume. The concentration of USP Amiloride Hydrochloride RS in the Standard solution is about 0.1 mg/mL. Sample solution: Transfer an equivalent to 5 mg of amiloride hydrochloride, from finely powdered Tablets (NLT 20), to a 50-mL volumetric flask containing 15.0 mL of methanol and 2.0 mL of 0.1 N hydrochloric acid. Sonicate for 10 min, dilute with water to volume, sonicate for an additional 10 min, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 286 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution (five replicate injections) Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H8ClN7O HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amiloride Hydrochloride RS (mg/mL) = nominal concentration of amiloride hydrochloride in the Sample solution (mg/mL)
Amiloride Hydrochloride Tablets

(Comment on this Monograph)id=m2660=Amiloride Hydrochloride Tablets=A-Monos.pdf) DEFINITION Amiloride Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C6H8ClN7O HCl. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. Thin-Layer Chromatography 201 Sample solution: Equivalent to 0.2 mg/mL of amiloride hydrochloride in methanol, from finely ground Tablets. Filter.
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90.0%110.0%
USP 32
IDENTIFICATION A. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution A: 0.2 mg/mL of USP Amiloride Hydrochloride RS in methanol Standard solution B: 2 mg/mL of USP Hydrochlorothiazide RS in methanol Sample solution: Equivalent to 0.2 mg/mL of amiloride hydrochloride, from ground Tablets in methanol and filter Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Tetrahydrofuran and 3 N ammonium hydroxide (22:3) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Develop the chromatogram until the solvent has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, air-dry, and examine under short-wavelength UV light. Acceptance criteria: The RF values of the amiloride hydrochloride and hydrochlorothiazide spots of the Sample solution correspond to those of the corresponding Standard solutions. ASSAY PROCEDURE Solution A: 136 g of monobasic potassium phosphate in 800 mL of water, and adjust with phosphoric acid to a pH of 3.0. Dilute with water to 1000 mL. Mobile phase: Methanol, water, and Solution A (25:71:4) Standard stock solution: 1.0 mg/mL of USP Amiloride Hydrochloride RS in methanol Standard solution: 0.1 mg/mL of USP Amiloride Hydrochloride RS and 1 mg/mL USP Hydrochlorothiazide RS, prepared by transfering 10.0 mL of Standard stock solution to a 100-mL volumetric flask containing 100 mg of USP Hydrochlorothiazide RS and 20.0 mL of methanol. Add 4.0 mL of 1 N hydrochloric acid, and dilute with water to volume. Sample solution: Transfer an equivalent to 5 mg of amiloride hydrochloridefrom powdered Tablets (NLT 20) to a 50-mL volumetric flask. Add 15.0 mL of methanol, and 2.0 mL of 1 N hydrochloric acid. Sonicate for 10 min, dilute with water to volume, sonicate for an additional 10 min, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 286 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for hydrochlorothiazide and amiloride hydrochloride are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between hydrochlorothiazide and amiloride hydrochloride
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 363 nm Sample solutions: Sample per Dissolution 711. Dilute with Medium as necessary. Standard solution: USP Amiloride Hydrochloride RS of known concentration in Medium [NOTEAn amount of methanol not to exceed 2% of the total volume of the Standard solution may be used to dissolve the amiloride hydrochloride.] Analysis Samples: Sample solution and Standard solution Tolerances: NLT 80% (Q) of the labeled amount of C6H8ClN7O HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905 Procedure for content uniformity Sample solution: Transfer 1 finely powdered Tablet to a 100-mL volumetric flask, add 60 mL of 0.1 N hydrochloric acid, and shake by mechanical means for 30 min. Dilute with 0.1 N hydrochloric acid to volume and centrifuge a portion of the mixture. Dilute an accurately measured portion of the clear supernatant quantitatively to obtain a solution containing 10 g/mL of amiloride hydrochloride. Standard solution: 10 g/mL of USP Amiloride Hydrochloride RS in the same medium Spectrometric conditions Analytical wavelength: 363 nm Blank: 0.1 N hydrochloric acid Analysis Samples: Sample solution and Standard solution Calculate the percentage of C6H8ClN7O HCl in the Tablet taken: Result = (AU/AS) (CS/CU) F (100/L) = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Amiloride Hydrochloride RS (g/mL) = concentration of the Sample solution (unit/mL) CU (unit=Tablet) F = conversion factor (g to mg) L = labeled quantity (mg) of amiloride hydrochloride in the Tablet Acceptance criteria: Meet the requriements AU AS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amiloride Hydrochloride RS
Amiloride Hydrochloride and Hydrochlorothiazide Tablets

(Comment on this Monograph)id=m2665=Amiloride Hydrochloride and Hydrochlorothiazide Tablets=A-Monos.pdf) DEFINITION Amiloride Hydrochloride and Hydrochlorothiazide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of amiloride hydrochloride (C6H8ClN7O HCl) and hydrochlorothiazide (C7H8ClN3O4S2).
USP 32
Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H8ClN7O HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response for amiloride hydrochloride from the Sample solution = peak response for amiloride hydrochloride from rS the Standard solution = concentration of USP Amiloride Hydrochloride CS RS, corrected for loss in weight in the Standard solution (mg/mL) = nominal concentration of amiloide hydrochloride CU in the Sample solution (mg/mL) Calculate the percentage of C7H8ClN3O4S2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of hydrochlorothiazide from the Sample solution rS = peak response of hydrochlorothiazide from the Standard solution CS = concentration of USP Hydrochlorothiazide RS in the Standard solution (mg/mL) CU = nominal concentration of hydrochlorothiazide in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amounts of C6H8ClN7O HCl and C7H8ClN3O4S2 PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Amiloride standard solution: 60 mg of USP Amiloride Hydrochloride RS (equivalent to 52 mg of anhydrous amiloride hydrochloride) in a 200-mL volumetric flask. Dissolve in and dilute with methanol to volume. Transfer 2.0 mL of this solution to a 100-mL volumetric flask, and dilute with Medium to volume. Hydrochlorothiazide standard solution: Transfer 100 mg of USP Hydrochlorothiazide RS to a 100-mL volumetric flask. Dissolve in and dilute with methanol to volume. Transfer 5.0 mL of this solution to a 100-mL volumetric flask, and dilute with Medium to volume. Transfer 10.0 mL of the resulting solution to a 50-mL volumetric flask, and dilute with Medium to volume. Sample solution A: Pass a portion of the solution under test through a 0.45-m glass fiber filter. Sample solution B: Transfer 5.0 mL of Sample solution A to a 25-mL volumetric flask, and dilute with Medium to volume. Detector: UV 363 nm for amiloride hydrochloride, 270 nm for hydrochlorothiazide Blank: Medium Analysis Samples: Amiloride standard solution, Hydrochlorothiazide standard solution, Sample solution A, and Sample solution B Calculate the percentage of C6H8ClN7O HCl dissolved: rU rU AUC AU270 AU363
Official Monographs / Amiloride 181

AS = absorbance of the Amiloride standard solution LC = tablet label claim of amiloride (mg) Correction for the interference of amiloride is made using the following equation:
= corrected absorbance of Sample solution A, 270 nm = absorbance of Sample solution B, 270 nm = absorbance of Sample solution A, 363 nm
ASAmiloride= absorbance of the Amiloride standard solution Calculate the amount of C7H8ClN3O4S2 dissolved, in percentage:
= corrected absorbance of Sample solution A, 270 nm = concentration of the Hydrochlorothiazide standard CS solution (mg/mL) 900 = volume of Medium (mL) (25/5) = dilution factor of Sample solution B 100 = conversion factor to percentage = absorbance of the Hydrochlorothiazide standard AS solution LC = tablet label claim of hydrochlorothiazide (mg) Tolerances: NLT 80% (Q) of the labeled amount of C6H8ClN7O HCl and NLT 75% (Q) of the labeled amount of C7H8ClN3O4S2 is dissolved. UNIFORMITY OF DOSAGE UNITS, Content Uniformity 905: Meet the requirements with respect to amiloride hydrochloride and hydrochlorothiazide AUC IMPURITIES Organic Impurities PROCEDURE Solution A, Mobile phase, and Sample solution: Proceed as directed in the Assay. Standard solution: 10 g/mL of USP Benzothiadiazine Related Compound A RS in the Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 286 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for hydrochlorothiazide and amiloride hydrochloride are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between hydrochlorothiazide and amiloride hydrochloride Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of benzothiadiazine related compound A in the portion of Tablets taken: Result = (rU/rS) (CS/CU) F 100
AU CS 900 100
= absorbance of Sample solution A = concentration of the Amiloride standard solution (mg/mL) = volume of Medium (mL) = conversion factor to percentage
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rU
USP 32
Carrier gas: Helium Flow rate: 6 mL/min Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.0 between the amiloxate peak and any other peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H20O3 in the portion of Amiloxate taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amiloxate RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% IMPURITIES Organic Impurities PROCEDURE Sample solution: Use the Sample solution as obtained in the Assay. Chromatographic system: Proceed as directed in the Assay. Injection size: 1 L System suitability Sample: Use the Standard solution prepared as directed in the Assay. Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Amiloxate taken: Result = (ri/rT) 100 = peak response for each impurity ri = sum of the responses of all the peaks rT Acceptance criteria Individual impurities: NMT 0.1% Total impurities: NMT 2.0% SPECIFIC TESTS SPECIFIC GRAVITY 841: 1.0371.041 REFRACTIVE INDEX 831: 1.5561.560 at 20 ACIDITY Sample solution: Transfer 50 mL of alcohol to a suitable container. Add 1 mL of phenolphthalein TS and sufficient 0.1 N sodium hydroxide to obtain a persistent pink color. Transfer 50 mL of this solution to a suitable container, and add 5.0 mL of Amiloxate. Analysis: Titrate with 0.1 N sodium hydroxide. Acceptance criteria: NMT 0.2 mL of titrant/mL of Amiloxate is required for neutralization. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amiloxate RS rU rS CS
= peak response of benzothiadiazine related compound A from the Sample solution = peak response of benzothiadiazine related rS compound A from the Standard solution = concentration of USP Benzothiadiazine Related CS Compound A RS in the Standard solution (g/mL) = nominal concentration of benzothiadiazine in CU the Sample solution (mg/mL) F = unit conversion factor, 0.001 mg/g Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amiloride Hydrochloride RS USP Benzothiadiazine Related Compound A RS USP Hydrochlorothiazide RS
Amiloxate
(Comment on this Monograph)id=m42600=Amiloxate=AMonos.pdf)
C15H20O3 248.32 4-Methoxycinnamic acid, isoamyl ester; 2-propenoic acid, 3-(4-methoxyphenyl)-3-methylbutyl ester [71617-10-2]. DEFINITION Amiloxate contains NLT 95.0% and NMT 105.0% of C15H20O3. IDENTIFICATION A. INFRARED ABSORPTION 197F B. ULTRAVIOLET ABSORPTION 197U Sample solution: 5.0 g/mL in alcohol Acceptance criteria: Absorptivities, calculated on the as-is basis, do not differ by more than 3.0%. ASSAY PROCEDURE Standard solution: 20 mg/mL of USP Amiloxate RS in tertbutyl methyl ether Sample solution: 20 mg/mL of Amiloxate in acetone Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.32-mm 25-m; coated with a 0.1-m film of G1 Temperature: See the temperature program table below.
Time (min) Injection port Detector Column 0 22.5 32.5 Temperature 240 260 60 240 240
USP 32
Official Monographs / Aminobenzoate 183

Concomitantly determine the absorbances of the solutions. Acceptance criteria: The absorbance of the solution obtained from the Sample solution does not exceed that of the solution obtained from the Standard solution, corresponding to NMT 0.002% of volatile diazotizable substances, as p-toluidine. SPECIFIC TESTS PH 791: 8.09.0, in a solution (1 in 20) LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Aminobenzoate Potassium RS
Aminobenzoate Potassium
(Comment on this Monograph)id=m2780=Aminobenzoate Potassium=A-Monos.pdf) DEFINITION Aminobenzoate Potassium contains NLT 98.5% and NMT 101.0% of C7H6KNO2, calculated on the dried basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Solution: 10 g/mL in 0.001 N sodium hydroxide B. Dissolve 400 mg in 10 mL of water, add 1 mL of 3 N hydrochloric acid, filter, and wash the precipitate with two 5-mL portions of cold water. Recrystallize from alcohol the precipitate so obtained, and dry at 110 for 1 h. The paminobenzoic acid so obtained melts between 186 and 189. C. A solution (1 in 100) meets the requirements of the flame test under Identification TestsGeneral 191, Potassium. ASSAY PROCEDURE Sample: 500 mg Analysis: Add 25 mL of water and 25 mL of 3 N hydrochloric acid, mix and cool in an ice bath. Titrate with 0.1 M sodium nitrite VS, using a calomel-platinum electrode system. Each mL of 0.1 M sodium nitrite is equivalent to 17.52 mg of C7H6KNO2. Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 1.4-g portion shows no more chloride than corresponds to 0.4 mL of 0.020 N hydrochloric acid (0.02%). CHLORIDE AND SULFATE, Sulfate 221: A 1.4-g portion shows no more sulfate than corresponds to 0.3 mL of 0.020 N sulfuric acid (0.02%). HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE: VOLATILE DIAZOTIZABLE SUBSTANCES Standard solution: Dissolve 10 mg of p-toluidine in 5 mL of methanol in a 100-mL volumetric flask, and add water to volume. Transfer 1 mL to a 100-mL volumetric flask, and dilute with water to volume. Sample solution: Transfer 5.0 g of Aminobenzoate Potassium to a suitable flask, and add a volume of 1.25 N sodium hydroxide that is just sufficient to dissolve the sample and to render the solution just alkaline to phenolphthalein TS. Dilute with water to 50 mL, and steam-distill the solution, collecting 95 mL of the distillate in a 100-mL volumetric flask. Add water to volume. Spectrometric conditions Mode: UV-Vis Analytical wavelength: 405 nm Analysis Samples: Standard solution and Sample solution Transfer 20.0-mL portions of the Samples to separate 100mL beakers, and transfer 20.0 mL of water to a third 100-mL beaker to provide the blank. Treat each as follows. Add 5.0 mL of 1 N hydrochloric acid, and cool in an ice bath. Add 2.0 mL of 0.1 M sodium nitrite dropwise, with stirring, allow to stand for 5 min for the diazotization reaction to be complete, add quickly to 10.0 mL of a cold solution of guaiacol (freshly prepared by dissolving 0.20 g of guaiacol in 100 mL of 1 N sodium hydroxide), and allow to stand for 30 min.
Aminobenzoate Potassium Capsules

(Comment on this Monograph)id=m2783=Aminobenzoate Potassium Capsules=A-Monos.pdf) DEFINITION Aminobenzoate Potassium Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of aminobenzoate potassium (C7H6KNO2). IDENTIFICATION Dissolve 1 g of the Capsule contents in 25 mL of water, add 5 mL of 3 N hydrochloric acid, and wash the precipitate with two 5-mL portions of cold water. Recrystallize from alcohol the precipitate so obtained, and dry at 110 for 1 h: the p-aminobenzoic acid so obtained melts between 186 and 189. ASSAY PROCEDURE Standard solution: 5 g/mL of USP Aminobenzoate Potassium RS Sample solution: Remove as completely as possible, and combine, the contents of NLT 20 Capsules. Transfer a portion of the combined contents, equivalent to 100 mg of aminobenzoate potassium, to a 200-mL volumetric flask, add 150 mL of water, shake by mechanical means for 30 min, dilute with water to volume, and filter. Pipet 2 mL of the filtrate into a 200-mL volumetric flask, and dilute with water to volume. Concomitantly determine the absorbances of the Standard solution and Sample solution. Spectrometric conditions Mode: UV Analytical wavelength: 270 nm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6KNO2 in the portion of Capsule contents taken: Result = (AU/AS) (CS/CU) F 100 AU AS CS CU F = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Aminobenzoate Potassium RS in the Standard solution (g/mL) = nominal concentration of aminobenzoate potassium in the Sample solution (mg/mL) = unit conversion factor, 0.001 mg/g
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USP 32
Aminobenzoate Potassium Tablets

(Comment on this Monograph)id=m2790=Aminobenzoate Potassium Tablets=A-Monos.pdf) DEFINITION Aminobenzoate Potassium Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aminobenzoate potassium (C7H6KNO2). IDENTIFICATION PROCEDURE Analysis: Dissolve 1 g of finely powdered Tablets in 25 mL of water, add 5 mL of 3 N hydrochloric acid, and wash the precipitate with two 5-mL portions of cold water. Recrystallize from alcohol the precipitate so obtained, and dry at 110 for 1 h. Acceptance criteria: The p-aminobenzoic acid so obtained melts between 186 and 189. ASSAY PROCEDURE Standard solution: 5 g/mL of USP Aminobenzoate Potassium RS Sample solution: Transfer an equivalent to 100 mg of aminobenzoate potassium, from finley powdered tablets (NLT 20), to a 200-mL volumetric flask, add 150 mL of water, shake by mechanical means for 30 min, dilute with water to volume and filter. Pipet 2 mL of the filtrate into a 200-mL volumetric flask, and dilute with water to volume. Spectrometric conditions Analytical wavelength: 270 nm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6KNO2 in the portion of Capsule contents taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Aminobenzoate Potassium RS in the Standard solution (g/mL) = nominal concentration of aminobenzoate CU potassium for the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% AU AS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Detector: UV 270 nm Standard solution: USP Aminobenzoate Potassium RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Tolerances: NLT 75% (Q) of the labeled amount of C7H6KNO2 is dissolved.
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Detector: UV 270 nm Standard solution: USP Aminobenzoate Potassium RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Tolerances: NLT 75% (Q) of C7H6KNO2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Aminobenzoate Potassium RS
Aminobenzoate Potassium for Oral Solution

(Comment on this Monograph)id=m2787=Aminobenzoate Potassium for Oral Solution=A-Monos.pdf) DEFINITION Aminobenzoate Potassium for Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of aminobenzoate potassium (C7H6KNO2). IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample Solution: 50 g/mL in water B. Dissolve 400 mg in 10 mL of water, add 1 mL of 3 N hydrochloric acid, filter, and wash the precipitate with two 5-mL portions of cold water. Recrystallize from alcohol the precipitate so obtained, and dry at 110 for 1 h: the paminobenzoic acid so obtained melts between 186 and 189. ASSAY PROCEDURE Sample: 100 mg Analysis: Add 5 mL of hydrochloric acid and 50 mL of water, cool to 15, and add 25 g of crushed ice. Titrate with 0.1 M sodium nitrite VS, using a calomelplatinum electrode system. Each mL of 0.1 M sodium nitrite is equivalent to 17.52 mg of C7H6KNO2. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for a solid packaged in single-unit containers MINIMUM FILL 755: Meets the requirements for a solid packaged in multiple-unit containers SPECIFIC TESTS PH 791: 7.09.0, in a solution (1 in 10) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aminobenzoate Potassium RS
USP 32
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Official Monographs / Aminobenzoic 185

20.0 mL of water to a third 100-mL beaker to provide the blank. Treat each as follows. Add 5.0 mL of 1 N hydrochloric acid, and cool in an ice bath. Add 2.0 mL of 0.1 M sodium nitrite dropwise, with stirring, allow to stand for 5 min for the diazotization reaction to be complete, add quickly to 10.0 mL of a cold solution of guaiacol (freshly prepared by dissolving 0.20 g of guaiacol in 100 mL of 1 N sodium hydroxide), and allow to stand for 30 min. Acceptance criteria: The absorbance of the solution obtained from the Sample solution does not exceed that of the solution obtained from the Standard solution, corresponding to NMT 0.002% of volatile diazotizable substances, as p-toluidine. SPECIFIC TESTS PH 791: 8.09.0, in a solution (1 in 20) LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Aminobenzoate Sodium RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Aminobenzoate Potassium RS
Aminobenzoate Sodium
(Comment on this Monograph)id=m2795=Aminobenzoate Sodium=A-Monos.pdf) DEFINITION Aminobenzoate Sodium contains NLT 98.5% and NMT 101.0% of C7H6NNaO2, calculated on the dried basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: 10 g/mL in 0.001 N sodium hydroxide B. Dissolve 400 mg in 10 mL of water, add 1 mL of 3 N hydrochloric acid, filter, and wash the precipitate with two 5-mL portions of cold water. Recrystallize from alcohol the precipitate so obtained, and dry at 110 for 1 h: the paminobenzoic acid so obtained melts between 186 and 189. C. IDENTIFICATION TESTSGENERAL, Sodium 191: A solution (1 in 100) meets the requirements of the flame test for sodium. ASSAY PROCEDURE Sample: 500 mg of Aminobenzoate Sodium Analysis: Add 25 mL of water and 25 mL of 3 N hydrochloric acid and cool in an ice bath. Titrate with 0.1 M sodium nitrite VS, using a calomel-platinum electrode system. Each mL of 0.1 M sodium nitrite is equivalent to 15.91 mg of C7H6NNaO2. Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 1.4-g portion shows no more chloride than corresponds to 0.4 mL of 0.020 N hydrochloric acid (0.02%). CHLORIDE AND SULFATE, Sulfate 221: A 1.4-g portion shows no more sulfate than corresponds to 0.3 mL of 0.020 N sulfuric acid (0.02%). HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE: VOLATILE DIAZOTIZABLE SUBSTANCES Blank: Water Standard stock solution: 10 mg of p-toluidine in 5 mL of methanol in a 100-mL volumetric flask, add water to volume Standard solution: Transfer 1 mL of Standard stock solution to a 100-mL volumetric flask, and dilute with water to volume. Sample solution: Transfer 5.0 g of Aminobenzoate Sodium to a suitable flask, and add a volume of 1.25 N sodium hydroxide that is just sufficient to dissolve the sample and to render the solution just alkaline to phenolphthalein TS. Dilute with water to 50 mL, and steam-distill the solution, collecting 95 mL of the distillate in a 100-mL volumetric flask. Add water to volume. Spectrometric conditions Mode: UV-Vis Analytical wavelength: At 405 nm Analysis Samples: Blank, Standard solution, and Sample solution Transfer 20.0-mL portions of the Standard solution and the Sample solution to separate 100-mL beakers, and transfer
Aminobenzoic Acid
(Comment on this Monograph)id=m2810=Aminobenzoic Acid=A-Monos.pdf)
C7H7NO2 Benzoic acid, 4-amino; p-Aminobenzoic acid [150-13-0].
137.14
DEFINITION Aminobenzoic Acid contains NLT 98.5% and NMT 101.5% of C7H7NO2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 5 g/mL in 0.001 N sodium hydroxide ASSAY PROCEDURE Sample: 250 mg of Aminobenzoic Acid Analysis: Proceed as directed under Nitrite Titration 451. Each mL of 0.1 M sodium nitrite is equivalent to 13.71 mg of C7H7NO2. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1: VOLATILE DIAZOTIZABLE SUBSTANCES Blank: water Standard stock solution: 10 mg of p-toluidine in 5 mL of methanol in a 100-mL volumetric flask, add water to volume Standard solution: Transfer 1 mL of Standard stock solution to a 100-mL volumetric flask, and dilute with water to volume.
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USP 32
Standard solution: Standard stock solution, Internal standard solution, and methanol (1:1:8). Pass through 0.6-m filter paper. Throughout the preparation, protect against actinic light. Sample solution: Gel, equivalent to 4.2 mg of aminobenzoic acid, to a 100-mL volumetric flask, and add 10.0 mL of Internal standard solution and 50 mL of methanol. Shake or sonicate, as necessary, and dilute with methanol to volume. Pass, if necessary, through filter paper (Whatman No. 41 or equivalent). Pass through 0.6-m filter paper. Throughout this preparation, protect against actinic light. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L11 Flow rate: 1 mL/min Injection size: 15 L System suitability Sample: Standard solution [NOTEChromatograph replicate 15-L injections of the Standard solution until the response ratio variability is within 1.0% of average.] [NOTEThe relative retention times of aminobenzoic acid and salicylic acid are about 1.0 and 3.0, respectively.] Suitability requirements Resolution: NLT 3.0 between aminobenzoic acid and salicylic acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H7NO2 in the portion of Gel taken: Result = (RU/RS) (CS/CU) 100 = ratios of the peak responses from the Sample solution RS = ratios of the peak responses from the Standard solution = concentration of USP Aminobenzoic Acid RS in CS the Standard solution (mg/mL) CU = nominal concentration for the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% OTHER COMPONENTS ALCOHOL DETERMINATION, Method II 611: (w/w) of C2H5OH 42.3%54.0% RU
Sample solution: Transfer 5.0 g of Aminobenzoic Acid to a suitable flask, and add a volume of 1.25 N sodium hydroxide that is just sufficient to dissolve the sample and to render the solution just alkaline to phenolphthalein TS. Dilute with water to 50 mL, and steam-distill the solution, collecting 95 mL of the distillate in a 100-mL volumetric flask. Add water to volume. Spectrometric conditions Mode: UV-Vis Cell: 1 cm Analytical wavelength: 450 nm Analysis Samples: Blank, Standard solution, and Sample solution Transfer 20.0-mL portions of the Standard solution and the Sample solution to separate 100-mL beakers, and transfer 20.0 mL of water to a third 100-mL beaker to provide the blank. Treat each as follows. Add 5.0 mL of 1 N hydrochloric acid, and cool in an ice bath. Add 2.0 mL of 0.1 M sodium nitrite dropwise, with stirring, allow to stand for 5 min for the diazotization reaction to be complete, add quickly to 10.0 mL of a cold solution of guaiacol (freshly prepared by dissolving 0.20 g of guaiacol in 100 mL of 1 N sodium hydroxide), and allow to stand for 30 min. Acceptance criteria: The absorbance of the solution obtained from the Sample solution does not exceed that of the solution obtained from the Standard solution, corresponding to NMT 0.002% of volatile diazotizable substances, as p-toluidine. PROCEDURE 2: ORDINARY IMPURITIES 466 Sample solution: Alcohol Standard solution: Alcohol Eluant: A mixture of toluene, ethyl acetate, and alcohol (60:20:20), in a nonequilibrated chamber Visualization: 1 SPECIFIC TESTS MELTING RANGE OR TEMPERATURE741: 186189 LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.2% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Aminobenzoic Acid RS
Aminobenzoic Acid Gel

(Comment on this Monograph)id=m2820=Aminobenzoic Acid Gel=A-Monos.pdf) DEFINITION Aminobenzoic Acid Gel contains NLT 90.0% and NMT 110.0% of the labeled amount of aminobenzoic acid (C7H7NO2). IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 5 g/mL in alcohol ASSAY PROCEDURE Mobile phase: Mix 300 mL of methanol and 10 mL of glacial acetic acid with 690 mL of water. Allow the mixture to cool, and pass, if necessary, through a suitable microporous membrane filter. Degas the solution. Internal standard solution: 7 mg/mL of salicylic acid in methanol. Dissolve by sonicating. Standard stock solution: 0.42 mg/mL of USP Aminobenzoic Acid RS in methanol
PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS PH 791: 4.06.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers USP REFERENCE STANDARDS 11 USP Aminobenzoic Acid RS
Aminobenzoic Acid Topical Solution

(Comment on this Monograph)id=m2830=Aminobenzoic Acid Topical Solution=A-Monos.pdf) DEFINITION Aminobenzoic Acid Topical Solution contains, in each mL, NLT 45 mg and NMT 55 mg of aminobenzoic acid (C7H7NO2).
USP 32
IDENTIFICATION A. To 1 mL of Topical Solution, add 1 mL of 1 N sodium hydroxide, and add, in order, 0.5 mL of potassium iodide TS, 0.5 mL of 3 N hydrochloric acid, and 0.5 mL of sodium hypochlorite TS: a brown precipitate is formed. B. To 1 mL of Topical Solution, add 2 mL of 3 N hydrochloric acid, and cool to about 10. Add 1 mL of 10 mg/mL sodium nitrite, then add a solution prepared by mixing 50 mg of 2-naphthol with 3 mL of 1.94 M sodium hydroxide: a red color is produced. ASSAY PROCEDURE Sample solution: 5 mL of Topical Solution Analysis: Transfer Sample solution to a suitable open vessel, evaporate on a steam bath to dryness, and proceed as directed under Nitrite Titration 451, beginning with Add 20 mL of hydrochloric acid. Each mL of 0.1 M sodium nitrite is equivalent to 13.71 mg of C7H7NO2. Acceptance criteria: 4555 mg/mL OTHER COMPONENTS ALCOHOL DETERMINATION 611: SPECIFIC TESTS SPECIFIC GRAVITY 841: 65%75% RU
Official Monographs / Aminocaproic 187

Mode: LC Detector: UV 210 nm Column: 4.6-mm 15-cm; packing L1 Column temperature: 30 Flow rate: 0.7 mL/min Injection size: 20 L [NOTERecord chromatograms for NLT two times the retention time of aminocaproic acid.] System suitability Sample: Standard solution [NOTEThe relative retention times for aminocaproic acid and methionine are about 0.76 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between aminocaproic acid and methionine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H13NO2 in the portion of Aminocaproic Acid taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of aminocaproic acid to the internal standard from the Sample solution = peak response ratio of aminocaproic acid to the RS internal standard from the Standard solution = concentration of USP Aminocaproic Acid in the CS Standard solution (mg/mL) = concentration of aminocaproic acid in the CU Sample solution (mg/mL) Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5%
0.8950.905
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.
Aminocaproic Acid
(Comment on this Monograph)id=m2880=Aminocaproic Acid=A-Monos.pdf)
C6H13NO2 Hexanoic acid, 6-amino-; 6-Aminohexanoic acid [60-32-2].
131.17
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at room temperature. USP REFERENCE STANDARDS 11 USP Aminocaproic Acid RS
DEFINITION Aminocaproic Acid contains NLT 98.5% and NMT 101.5% of C6H13NO2, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Solution A: 0.55 mg/mL of sodium 1-heptanesulfonate Mobile phase: Dissolve 10 g of monobasic potassium phosphate in 300 mL of Solution A, add 250 mL of methanol, followed by another 300 mL of Solution A. Adjust the mixture with phosphoric acid to a pH of 2.2, and dilute with Solution A to 1 L. Internal standard solution: 1.25 mg/mL of methionine Standard stock solution: 12.5 mg/mL of USP Aminocaproic Acid RS Standard solution: Standard stock solution, Internal standard solution, and water (5:2:93) Sample stock solution: 12.5 mg/mL of Aminocaproic Acid Sample solution: Sample stock solution, Internal standard solution, and water (5:2:93) Chromatographic system (See Chromatography 621, System Suitability.)
Aminocaproic Acid Injection

(Comment on this Monograph)id=m2910=Aminocaproic Acid Injection=A-Monos.pdf) DEFINITION Aminocaproic Acid Injection is a sterile solution of Aminocaproic Acid in Water for Injection. It contains NLT 95.0% and NMT 107.5% of the labeled amount of aminocaproic acid (C6H13NO2). IDENTIFICATION INFRARED ABSORPTION 197K Sample: Mix 2 mL of Injection, added dropwise, with 100 mL of acetone, rapidly stirring the mixture with a glass rod to induce crystallization. Allow the mixture to stand for 15 min, and pass through a medium-porosity, sintered-glass filter. Wash the crystals with 25 mL of acetone, apply a vacuum to remove the solvent, dry at 105 for 30 min, and cool: the residue is used as the Sample. ASSAY PROCEDURE Mobile phase: Transfer 11 g of sodium 1-pentanesulfonate and 40 g of anhydrous sodium sulfate to a 2-L volumetric
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1 N hydrochloric acid in a 100-mL beaker. Decant and discard the hydrochloric acid, and wash the resin with five 10-mL portions of water, decanting and discarding the liquid following each washing. Analysis: Place the washed resin in a glass-stoppered, conical flask, and add a volume of Oral Solution, equivalent to 250 mg of aminocaproic acid, and 10 mL of water. Insert the stopper in the flask, and shake by mechanical means for 30 min. Transfer the resin slurry to a medium-porosity, sintered-glass funnel, wash with 100 mL of water, apply suction to filter, and discard the washing. Place a beaker under the stem of the funnel, add 10 mL of 1 N hydrochloric acid to the resin, stir for 45 min, and filter by applying suction. Evaporate the filtrate on a steam bath to dryness, dry at 105 for 1 h, and cool. Acceptance criteria: The residue so obtained meets the requirements for the test. ASSAY PROCEDURE Sample: Equivalent to 250 mg of aminocaproic acid Analysis: Add 80 mL of glacial acetic acid and 10 drops of a solution (1 in 500) of crystal violet in chlorobenzene. Titrate with 0.1 N perchloric acid in dioxane VS to a blue endpoint. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 13.12 mg of C6H13NO2. Acceptance criteria: 95.0%115.0% SPECIFIC TESTS PH 791: 6.06.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aminocaproic Acid RS
flask, and dissolve in 500 mL of water. Add 20 mL of 1 N sulfuric acid and 30 mL of acetonitrile, and dilute with water to volume. Standard solution: 2.5 mg/mL of USP Aminocaproic Acid RS in Mobile phase System suitability solution: Mix 20 L of benzyl alcohol with 100 mL of water. Dilute 1.0 mL of this solution with the Standard solution to 10 mL. Sample stock solution: Equivalent to 25 mg/mL of aminocaproic acid, from Injection in water Sample solution: 2.5 mg/mL of Sample stock solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: NLT 7.0 between benzyl alcohol and aminocaproic acid in the System suitability solution [NOTEThe aminocaproic acid peak elutes prior to the benzyl alcohol peak.] Relative standard deviation: NMT 1.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H13NO2 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Aminocaproic Acid RS in the Standard solution (mg/mL) CU = nominal concentration of aminocaproic acid in the Sample solution (mg/mL) Acceptance criteria: 95.0%107.5% rU rS CS SPECIFIC TESTS PH 791: 6.07.6 OTHER REQUIREMENTS: It meets the requirements under Injections 1. BACTERIAL ENDOTOXINS TEST 85: NMT 0.05 USP Endotoxin Unit/mg of aminocaproic acid ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Aminocaproic Acid RS USP Endotoxin RS
Aminocaproic Acid Tablets

(Comment on this Monograph)id=m2950=Aminocaproic Acid Tablets=A-Monos.pdf) DEFINITION Aminocaproic Acid Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of aminocaproic acid (C6H13NO2). IDENTIFICATION INFRARED ABSORPTION 197K Sample solution: Triturate 2 Tablets with 10 mL of water, and filter into 100 mL of acetone. Swirl the mixture, and allow to stand for 15 min to complete crystallization. Pass through a medium-porosity, sintered-glass filter, and wash the crystals with 25 mL of acetone. Apply vacuum to remove the solvent, then dry at 105 for 30 min, and cool: the residue is used as the sample. ASSAY PROCEDURE Sample solution: Equivalent to 5 mg/mL of aminocaproic acid from powdered Tablets (NLT 20) in glacial acetic acid. Heat gently to effect solution, and cool. Analysis: To 100 mL of Sample solution, add 10 drops of a solution (1 in 500) of crystal violet in chlorobenzene and titrate with 0.1 N perchloric acid in dioxane VS to a blue endpoint. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 13.12 mg of (C6H13NO2).
Aminocaproic Acid Oral Solution

(Comment on this Monograph)id=m2940=Aminocaproic Acid Oral Solution=A-Monos.pdf) DEFINITION Aminocaproic Acid Oral Solution contains NLT 95.0% and NMT 115.0% of the labeled amount of aminocaproic acid (C6H13NO2). IDENTIFICATION INFRARED ABSORPTION 197K Sample: Mix 1 g of ion-exchange resin (strongly acidic styrene-divinylbenzene cation-exchange resin) with 10 mL of
USP 32
Official Monographs / Aminoglutethimide 189

and add 500 mL of water. Adjust by the addition of either 1 N acetic acid or 1 N potassium hydroxide to a pH of 5.0 0.1. Dilute with water to volume. Mobile phase: Methanol and Solution A (27:73) Diluent: Methanol and Solution A (1:1) Standard solution: 0.5 mg/mL of USP Aminoglutethimide RS in Diluent Sample solution: 0.5 mg/mL of Aminoglutethimide in Diluent [NOTEPass through a 0.45-m or finer porosity filter, discarding the first 5 mL of the filtrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 3.9-mm 15-cm; 4-m packing L1 Column temperature: 40 Flow rate: 1.3 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.7 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C13H16N2O2 in the portion of Aminoglutethimide taken: Result = (rU/rS) (CS/CU) 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Aminoglutethimide RS in the Standard solution (mg/mL) CU = concentration of Aminoglutethimide in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE 1: LIMIT OF AZO-AMINOGLUTETHIMIDE [NOTEUse low-actinic glassware. Conduct this test promptly under subdued light. Wear protective gloves resistant to dimethyl sulfoxide to prevent contact with skin. Use shaking, not sonication or heat, to dissolve the USP Azo-aminoglutethimide RS and the Sample.] Solution A: 150 mL of 0.1 N acetic acid and 50 mL of 0.1 N potassium hydroxide, diluted in water to 1000 mL Mobile phase: 100 mg of edetate disodium in 350 mL of Solution A, add 650 mL of methanol and cool to room temperature. Adjust with glacial acetic acid to a pH of 5.0 0.1. Standard solution: 0.5 g/mL of USP Azoaminoglutethimide RS in dimethyl sulfoxide Sample solution: 1 mg/mL of Aminoglutethimide in dimethyl sulfoxide Chromatographic system (See Chromatography 621, System Suitability.)
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min pH 9.5 borate buffer: Dissolve 6.185 g of boric acid and 7.930 g of potassium chloride in 1000 mL of water, and add 60 mL of 1.0 N sodium hydroxide. Dilute with water to 2000 mL, and add 1.0 N sodium hydroxide, if necessary, to adjust to a pH of 9.5 0.1. Standard solution: 0.5 mg/mL of USP Aminocaproic Acid RS in water Analysis: Into 3 separate 50-mL volumetric flasks pipet (a) 1 mL of a filtered portion of the solution under test, (b) 1 mL of the Standard solution, and (c) 1 mL of water to provide a blank. Add 20.0 mL of pH 9.5 borate buffer and 3.0 mL of freshly prepared -naphthoquinone-4-sodium sulfonate solution (1 in 500) to each, swirl to mix, and place the 3 flasks in a water bath maintained at a temperature of 65 5 for 45 min. Cool, and dilute each with water to volume. Determine the amount of C6H13NO2 dissolved from absorbances, at the wavelength of maximum absorbance at about 460 nm, obtained from the Sample solution in comparison with those obtained from the Standard solution, using the blank to set the instrument. Tolerances: NLT 75% (Q) of the labeled amount of C6H13NO2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aminocaproic Acid RS
Aminoglutethimide
(Comment on this Monograph)id=m2975=Aminoglutethimide=A-Monos.pdf)
C13H16N2O2 2,6-Piperidinedione, 3-(4-aminophenyl)-3-ethyl-; 2-(p-Aminophenyl)-2-ethylglutarimide [125-84-8].
232.28
DEFINITION Aminoglutethimide contains NLT 98.0% and NMT 102.0% of C13H16N2O2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 242 nm Medium: Methanol Solution: 10 g/mL Acceptance criteria: Absorptivities differ by NMT 2.0% ASSAY PROCEDURE Solution A: Add 240 mL of 0.1 N acetic acid to 200 mL of 0.1 N potassium hydroxide in a 2000-mL volumetric flask,
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USP 32
Acceptance criteria: No turbidity is produced. PH 791 Sample solution: 1 mg/mL in dilute methanol (1 in 20) Acceptance criteria: 6.27.3 LOSS ON DRYING 731: Dry at 105 to constant weight: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Aminoglutethimide RS USP m-Aminoglutethimide RS USP Azo-aminoglutethimide RS
Mode: LC Detector: UV 328 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.2 Capacity factor: 2.05.0 Column efficiency: NLT 800 theoretical plates Analysis Samples: Standard solution and Sample solution [NOTEThe aminoglutethimide elutes with the dimethyl sulfoxide.] Calculate the percentage of azo-aminoglutethimide in the specimen of Aminoglutethimide taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Azo-aminoglutethimide RS in the Standard solution (g/mL) = concentration of Aminoglutethimide in the CU Sample solution (mg/mL) Acceptance criteria: NMT 0.03% of 3,3-(azodi-4,1phenylene)-3,3-dimethylbis-[2,6-piperidinedione] (corresponding to azo-aminoglutethimide) PROCEDURE 2: LIMIT OF m-AMINOGLUTETHIMIDE Solution A, Mobile phase, Diluent, and Chromatographic system: Prepare as directed in the Assay. Standard solution: 10 g/mL of USP mAminoglutethimide RS in Diluent Sample solution: 1 mg/mL of Aminoglutethimide in Diluent [NOTEPass through a 0.45-m or finer porosity filter, discarding the first 5 mL of the filtrate] Analysis Samples: Standard solution and Sample solution [NOTEThe relative retention times for aminoglutethimide and m-aminoglutethimide are about 0.8 and 1.0, respectively.] Calculate the percentage of m-aminoglutethimide in the specimen of Aminoglutethimide taken: rU rS CS Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP m-Aminoglutethimide RS in the Standard solution (mg/mL) = concentration of Aminoglutethimide in the CU Sample solution (mg/mL) Acceptance criteria: NMT 2.0% of m-aminoglutethimide is found. Calculate the percentage of each peak, other than the main peak and the m-aminoglutethimide peak, if present, by the same formula: rU rS CS Result = (rU/rT) 100 = response of each peak = sum of the responses of all the peaks in the chromatogram of the Sample solution Acceptance criteria: NMT 1.0% total impurities, other than m-aminoglutethimide rU rT SPECIFIC TESTS SULFATE Sample solution: 1 mg/mL in dilute methanol (1 in 20) Analysis: To 100 mL of Sample solution, add 1.0 mL of 3 N hydrochloric acid and 2.0 mL of barium chloride TS
Aminoglutethimide Tablets
(Comment on this Monograph)id=m2985=Aminoglutethimide Tablets=A-Monos.pdf) DEFINITION Aminoglutethimide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aminoglutethimide (C13H16N2O2). IDENTIFICATION INFRARED ABSORPTION 197M Sample: Transfer 500 mg of finely powdered Tablets to a suitable container, add 25 mL of acetone, mix, and filter. Evaporate the filtrate at room temperature to dryness, and dry the residue in vacuum over silica gel for 2 h. ASSAY PROCEDURE Acetate buffer: Prepare a solution in water, containing 120 mL of 0.1 N acetic acid and 100 mL of 0.1 N potassium hydroxide/L of buffer. Before final dilution, adjust by the addition of either 1 N acetic acid or 1 N potassium hydroxide to a pH of 5.0 0.1. Mobile phase: Methanol and Acetate buffer (27:73) Diluent: Methanol and Acetate buffer (1:1) Standard solution: 0.5 mg/mL of USP Aminoglutethimide RS in Diluent Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 200 mg of aminoglutethimide, to a 200-mL volumetric flask. Add 130 mL of Diluent, and sonicate for 5 min. Shake by mechanical means for 30 min, and dilute with Diluent to volume. Centrifuge this solution, transfer 25.0 mL of the clear supernatant to a 50-mL volumetric flask, dilute with Diluent to volume and pass through a 0.45-m or finer porosity filter, discarding the first 5 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 3.9-mm 15-cm; 4-m packing L1 Column temperature: 40 Flow rate: 1.3 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.7 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C13H16N2O2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100
USP 32
= peak area from the Sample solution = peak area from the Standard solution = concentration of USP Aminoglutethimide RS in the Standard solution (mg/mL) CU = nominal concentration of aminoglutethimide in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Dilute hydrochloric acid (7 in 1000); 1000 mL Apparatus 1: 100 rpm Time: 30 min Detector: UV 237 nm Sample solution: Sample per Dissolution 711. Dilute with pH 7.5 phosphate buffer to a concentration that is similar to the Standard solution. Standard solution: USP Aminoglutethimide RS in a mixture of dilute hydrochloric acid and pH 7.5 phosphate buffer, having a ratio similar to the Sample solution Tolerances: NLT 70% (Q) of the labeled amount of C13H16N2O2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Buffer: Prepare a solution in water, containing 120 mL of 0.1 N acetic acid and 100 mL of 0.1 N potassium hydroxide/L of buffer. Before final dilution, adjust by the addition of either 1 N acetic acid or 1 N potassium hydroxide to a pH of 5.0 0.1. Mobile phase: Methanol and Buffer (27:73) Diluent: Methanol and Buffer (1:1) Standard solution: 10 g/mL of USP mAminoglutethimide RS in Diluent Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 200 mg of aminoglutethimide, to a 200-mL volumetric flask. Add 130 mL of Diluent, and sonicate for 5 min. Shake by mechanical means for 30 min, dilute with Diluent to volume, and pass through a 0.45-m or finer porosity filter, discarding the first 5 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 3.9-mm 15-cm; 4-m packing L1 Column temperature: 40 Flow rate: 1.3 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.7 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution [NOTEThe relative retention times for aminoglutethimide and m-aminoglutethimide are 0.8 and 1.0, respectively.] Calculate the percentage of each peak, other than the main peak and the m-aminoglutethimide peak, if present: Result = (rU/rT) 100 rU rT = response of each impurity peak in the Sample solution = sum of the responses of all of the peaks excluding that of the m-aminoglutethimide peak in the Sample solution rU rS CS
Official Monographs / Aminohippurate 191

Acceptance criteria: NMT 2.0% total impurities, other than m-aminoglutethimide, is found. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Aminoglutethimide RS USP m-Aminoglutethimide RS
Aminohippurate Sodium Injection

(Comment on this Monograph)id=m3000=Aminohippurate Sodium Injection=A-Monos.pdf)
C9H9N2NaO3 Glycine, N-(4-aminobenzoyl)-, monosodium salt; Monosodium p-aminohippurate [94-16-6].
216.17
DEFINITION Aminohippurate Sodium Injection is a sterile solution of Aminohippuric Acid in Water for Injection prepared with the aid of Sodium Hydroxide. It contains NLT 95.0% and NMT 105.0% of the labeled amount of C9H9N2NaO3. IDENTIFICATION A. Procedure Analysis: Dilute a volume of Injection, equivalent to 100 mg of aminohippuric acid, to 50 mL and acidify with hydrochloric acid. Add 0.5 mL of 3 N hydrochloric acid, 0.5 mL of sodium nitrite solution (1 in 10), and a solution of 0.20 g of 2-naphthol in 10 mL of 6 N ammonium hydroxide. Acceptance criteria: A red color is produced. B. Procedure Analysis: Transfer a volume of Injection, equivalent to 200 mg of aminohippurate sodium, to a test tube, and add, in the order named, 2 mL of potassium iodide TS, 10 mL of water, and 5 mL of sodium hypochlorite TS. Acceptance criteria: A red color is produced. C. Identification TestsGeneral, Sodium 191: Meets the requirements ASSAY PROCEDURE Sample: Equivalent to 1 g of aminohippurate sodium Analysis: Transfer to a 200-mL volumetric flask, and dilute with water to volume. Transfer 50.0 mL of the solution to a suitable container, add 5 mL of hydrochloric acid, stir, cool to 15, slowly titrate with 0.1M sodium nitrate VS, and proceed as directed under Nitrite Titration 451, beginning with Determine the endpoint. Each mL of 0.1 M sodium nitrite is equivalent to 19.42 mg of C9H10N2O3. Acceptance criteria: 95.0%105.0% SPECIFIC TESTS PH 791: 6.77.6 OTHER REQUIREMENTS: Meets the requirements under Injections 1 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.04 USP Endotoxin Unit/mg of aminohippurate sodium.
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USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Aminohippuric Acid RS
Aminopentamide Sulfate
(Comment on this Monograph)id=m3040=Aminopentamide Sulfate=A-Monos.pdf)
Aminohippuric Acid
(Comment on this Monograph)id=m3030=Aminohippuric Acid=A-Monos.pdf) 394.49 C19H24N2O H2SO4 -[2-(Dimethylamino)propyl]--phenylbenzeneacetamide sulfate [60-46-8]. DEFINITION Aminopentamide Sulfate contains NLT 95.0% and NMT 103.0% of C19H24 N2O H2SO4. C9H10N2O3 Glycine, N-(4-aminobenzoyl)-; p-Aminohippuric acid [61-78-9]. 194.19 IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sulfate 191 ASSAY PROCEDURE Sample: 500 mg of Aminopentamide Sulfate Analysis: Add Sample to 100 mL of dimethylformamide in a suitable container, add 5 drops of thymol blue TS, and titrate with 0.1 N lithium methoxide VS in toluene to a deep blue endpoint. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N lithium methoxide is equivalent to 19.72 mg of C19H24N2O H2SO4. Acceptance criteria: 95.0%103.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
DEFINITION Aminohippuric Acid contains NLT 98.0% and NMT 100.5% of C9H10N2O3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Analysis Dissolve 10 mg in 5 mL of water, and add 0.5 mL of 3 N hydrochloric acid, 0.5 mL of sodium nitrite solution (1 in 10), and a solution of 0.20 g of 2-naphthol in 10 mL of 6 N ammonium hydroxide. Acceptance criteria: A red color is produced. ASSAY PROCEDURE Sample: 150 mg of Aminohippuric Acid Analysis: To the Sample, add 5 mL of hydrochloric acid and 50 mL of water, stir until dissolved, cool to 15, and slowly titrate with 0.1 M sodium nitrate VS, and proceed as directed under Nitrite Titration 451, beginning with Determine the endpoint. Each mL of 0.1 M sodium nitrite is equivalent to 19.42 mg of C9H10N2O3. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.25% HEAVY METALS, Method II 231: NMT 10 ppm SPECIFIC TESTS LOSS ON DRYING 731: 0.25% of its weight. Dry at 105 for 2 h: it loses NMT
NMT 0.5%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 179186 PH 791 Sample solution: 25 mg/mL Acceptance criteria: 1.23.0 LOSS ON DRYING 731: Dry at 105 for 4 h: it loses NMT 4.4% of its weight. CLARITY AND COLOR OF SOLUTION: Dissolve 500 mg in 10 mL of water: the solution is clear and colorless. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Aminopentamide Sulfate RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Aminohippuric Acid RS
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PH 791: 2.54.5 OTHER REQUIREMENTS: Meets the requirements under Injections 1 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 25 USP Endotoxin Units/mg of aminopentamide sulfate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose or multiple-dose , as described under Injections 1, Containers for Injections. Store at controlled room temperature. LABELING: Label Injection to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Aminopentamide Sulfate RS USP Endotoxin RS
Aminopentamide Sulfate Injection

(Comment on this Monograph)id=m3045=Aminopentamide Sulfate Injection=A-Monos.pdf) DEFINITION Aminopentamide Sulfate Injection is a sterile solution of Aminopentamide Sulfate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of aminopentamide sulfate (C19H24N2O H2SO4). IDENTIFICATION PROCEDURE Sample: Transfer 10 mL of the Injection to a separator, add sodium hydroxide TS until alkaline to litmus, and extract with 25 mL of chloroform. Transfer a few drops of the chloroform extract to a KRS-5 plate, and allow to dry. Record the infrared absorption spectrum by the attenuated total reflectance technique (See Spectrophotometry and LightScattering 851). Acceptance criteria: The spectrum thus obtained exhibits maxima only at the same wavelengths as that of a similar preparation of USP Aminopentamide Sulfate RS, concomitantly measured. ASSAY PROCEDURE Solution A: 28.8 mg/mL of sodium lauryl sulfate in glacial acetic acid and water (1:4). Pass through a filter having a 0.5-m or finer porosity. Mobile phase: Acetonitrile, methanol, Solution A, and water (7:7:1:5) Standard solution: USP Aminopentamide Sulfate RS in water, at a concentration equivalent to the labeled concentration of aminopentamide sulfate Sample solution: Use the undiluted Injection. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.5 Relative standard deviation: NMT 2% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C19H24N2O H2SO4 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response in the Sample solution = peak response in the Standard solution = concentration of USP Aminopentamide Sulfate RS in the Standard solution (mg/mL) = nominal concentration of aminopentamide CU sulfate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements under Test for Sterility of the Product to be Examined, Membrane Filtration
Aminopentamide Sulfate Tablets

(Comment on this Monograph)id=m3050=Aminopentamide Sulfate Tablets=A-Monos.pdf) DEFINITION Aminopentamide Sulfate Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of aminopentamide sulfate (C19H24N2O H2SO4). IDENTIFICATION A. PROCEDURE Sample: Transfer powdered Tablets, equivalent to 2 mg of aminopentamide, to a separator, add 20 mL of water and 3 mL of 10 N sodium hydroxide. Extract with two 20-mL portions of methylene chloride, and evaporate the combined methylene chloride extracts to a volume of about 0.5 mL. Transfer a few drops of the chloroform concentrate to a KRS-5 plate, and allow to dry. Record the IR absorption spectrum by the attenuated total reflectance technique (see Spectrophotometry and Light-Scattering 851). Acceptance criteria: The spectrum thus obtained exhibits maxima only at the same wavelengths as that of a similar preparation of USP Aminopentamide Sulfate RS, concomitantly measured. ASSAY PROCEDURE Solution A: 28.8 mg/mL of sodium lauryl sulfate in glacial acetic acid and water (1:4). Pass through a filter having a 0.5-m or finer porosity. Mobile phase: Acetonitrile, methanol, Solution A, and water (7:7:1:5) Standard solution: 0.02 mg/mL of USP Aminopentamide Sulfate RS in Mobile phase Sample solution: Equivalent to 0.02 mg/mL of aminopentamide, from Powdered Tablets in Mobile phase [NOTEFinely powder NLT 10 Tablets. After adding Mobile phase sonicate for 5 min, and stir by mechanical means for 10 min. Pass this mixture through a filter having a 0.5-m or finer porosity, discarding the first 5 mL of the filtrate. Use the clear filtrate.] Chromatographic system (See Chromatography 621, System Suitability.)
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anhydrous theophylline (C7H8N4O2), calculated on the anhydrous basis.
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Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Temperature: 40 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 900 theoretical plates Relative standard deviation: NMT 2% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C19H24N2O H2SO4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response of the Sample solution = peak responses of the Standard solution = concentration of USP Aminopentamide Sulfate RS in Standard solution (mg/mL) = nominal concentration of aminopentamide sulfate in the Sample solution (mg/mL)
SPECIFIC TESTS LOSS ON DRYING 731: Dry 1 g of powdered Tablets in a vacuum at a pressure of 5 mm of mercury or less at 60 for 3 h: it loses NMT 4.0% of its weight. PERFORMANCE TESTS DISINTEGRATION 701 Medium: Simulated gastric fluid TS Time: NMT 10 min UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label Tablets to indicate that they are for veterinary use only. USP REFERENCE STANDARDS 11 USP Aminopentamide Sulfate RS
IDENTIFICATION Sample 1: Dissolve 500 mg in 20 mL of water, add, with constant stirring, 1 mL of 3 N hydrochloric acid, and filter (retain the filtrate). Sample 2: Wash the precipitate from Sample 1 with small portions of cold water, and dry at 105 for 1 h. A. Sample 2 melts between 270 and 274. B. PROCEDURE Analysis: To 10 mg of Sample 2, in a porcelain dish, add 1 mL of hydrochloric acid and 100 mg of potassium chlorate, evaporate on a steam bath to dryness, and invert the dish over a vessel containing a few drops of 6 N ammonium hydroxide. Acceptance criteria: The residue acquires a purple color, which is destroyed by solutions of fixed alkalies. C. PROCEDURE Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide to render alkaline, shake by mechanical means for 10 min, add 5 mL of 3 N hydrochloric acid to acidify, chill, collect the precipitated disulfonamide of ethylenediamine, wash with water, recrystallize from water, and dry at 105 for 1 h. Acceptance criteria: The dried precipitate melts between 164 and 171. ASSAY PROCEDURE Mobile phase: 200 mL of methanol, 960 mg of sodium 1pentanesulfonate, and sufficient water to make 1 L. Adjust with glacial acetic acid to a pH of 2.9 0.1. Diluent: Methanol and water (1:4) Standard solution: 80 g/mL of USP Theophylline RS in Diluent Solution A: 80 g/mL of theobromine in Standard solution System suitability solution: 64 g/mL of theophylline and 64 g/mL of theobromine, from Solution A in Diluent Sample solution: 96 g/mL of Aminophylline in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for theobromine and theophylline are 0.65 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between theobromine and theophylline, System suitability solution Tailing factor: NMT 2.0 for theophylline peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H8N4O2 in the portion of Aminophylline taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Theophylline RS in the Standard solution (mg/mL) = concentration of aminophylline in the Sample solution (mg/mL)
Aminophylline
(Comment on this Monograph)id=m3080=Aminophylline=AMonos.pdf) Change to read:
C16H24N10O4 (anhydrous) 420.43 1H-Purine-2,6-dione, 3,7-dihydro-1,3-dimethyl-, compd. with 1,2-ethanediamine (2:1); Theophylline compound with ethylenediamine (2:1) [317-34-0]. Dihydrate 456.46 [v5897-66-5vUSP32]. DEFINITION Aminophylline is anhydrous or contains NMT two molecules of water of hydration. It contains NLT 84.0% and NMT 87.4% of
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Acceptance criteria: The residue acquires a purple color, which is destroyed by solutions of fixed alkalies. ASSAY PROCEDURE Mobile phase: 200 mL of methanol, 960 mg of sodium 1pentanesulfonate, and sufficient water to make 1 L. Adjust with glacial acetic acid to a pH of 2.9 0.1. Diluent: Methanol and water (1:4) Standard solution: 0.08 mg/mL of USP Theophylline RS in Diluent Solution A: 0.08 mg/mL of theobromine in the Standard solution System suitability solution: 64 g/mL of theophylline and 64 g/mL of theobromine, from Solution A in Diluent Sample solution: Equivalent to 0.08 mg/mL of theophylline, from Injection in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for theobromine and theophylline are about 0.65 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between theobromine and theophylline, System suitability solution Tailing factor: NMT 2.0 for the theophylline peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of C7H8N4O2 in the portion of Injection taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response of the Sample solution = peak response of the Standard solution = concentration of the USP Theophyline RS in the Standard solution (mg/mL) = nominal concentration of aminophylline in the Sample solution (mg/mL)
OTHER COMPONENTS ETHYLENEDIAMINE CONTENT Sample solution: 16.67 mg/mL Analysis: To 30 mL of Sample solution, add methyl orange TS and titrate with 0.1 N hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is equivalent to 3.005 mg of C2H8N2. Acceptance criteria: The content of ethylenediamine (C2H8N2) is between 157 mg and 175 mg/g of C7H8N4O2 found in the Assay. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
NMT 0.15%
SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.75% (anhydrous form) and NMT 7.9% (hydrous form), determined on 1.5 g, a mixture of 25 mL of chloroform and 25 mL of methanol being used in place of the methanol solvent. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate whether it is anhydrous or hydrous and also to state the content of anhydrous theophylline. USP REFERENCE STANDARDS 11 USP Theophylline RS
Aminophylline Injection
(Comment on this Monograph)id=m3130=Aminophylline Injection=A-Monos.pdf) DEFINITION Aminophylline Injection is a sterile solution of Aminophylline in Water for Injection, or is a sterile solution of Theophylline in Water for Injection prepared with the aid of Ethylenediamine. It contains, in each mL, an amount of aminophylline equivalent to NLT 93.0% and NMT 107.0% of the labeled amount of anhydrous theophylline (C7H8N4O2). Aminophylline Injection may contain an excess of Ethylenediamine, but no other substance may be added for the purpose of pH adjustment. [NOTEDo not use the Injection if crystals have separated.] IDENTIFICATION Sample 1: Dilute an equivalent of 500 mg of aminophylline, with water to 20 mL, and add, with constant stirring, 1 mL of 3 N hydrochloric acid or enough to precipitate the theophylline completely, and filter. Sample 2: Wash the precipitate from Sample 1 with a small portion of cold water, and dry at 105 for 1 h. A. PROCEDURE Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide to render alkaline. Shake by mechanical means for 10 min, add 5 mL of 3 N hydrochloric acid to acidify, and chill. Collect the precipitated disulfonamide of ethylenediamine, wash with water, recrystallize from water, and dry at 105 for 1 h. Acceptance criteria: Melts between 164 and 171 B. PROCEDURE: Sample 2 melts between 270 and 274. C. PROCEDURE Analysis: To 10 mg of Sample 2 contained in a porcelain dish, add 1 mL of hydrochloric acid and 100 mg of potassium chlorate, evaporate on a steam bath to dryness, and invert the dish over a vessel containing a few drops of 6 N ammonium hydroxide.
OTHER COMPONENTS CONTENT OF ETHYLENEDIAMINE Sample solution: Dilute an equivalent to 500 mg of Aminophylline to 30 mL, if necessary. Analysis: Add methyl orange TS and titrate with 0.1 N hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is equivalent to 3.005 mg of C2H8N2. Acceptance criteria: 166192 mg of ethylenediamine (C2H8N2) per g of C7H8N4O2 found in the Assay. SPECIFIC TESTS PH 791: 8.69.0 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: Meets the requirements under Injections 1 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.0 USP Endotoxin Unit/mg of aminophylline.
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Tailing factor: NMT 2.0 for theophylline peak, System suitability solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of C7H8N4O2 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Theophylline RS in the Standard solution (mg/mL) = nominal concentration of theophylline in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF ETHYLENEDIAMINE Sample solution: Equivalent to 500 mg of aminophylline to 30 mL, dilute with water if necessary. Analysis: Add methyl orange TS, and titrate with 0.1 N hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is equivalent to 3.005 mg of C2H8N2. Acceptance criteria: 176283 mg of ethylenediamine (C2H8N2) per g of C7H8N4O2 found in the Assay SPECIFIC TESTS PH 791: 8.59.7 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Oral Solution to state the content of anhydrous theophylline. USP REFERENCE STANDARDS 11 USP Theophylline RS rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers from which carbon dioxide has been excluded, preferably of Type I glass, protected from light. LABELING: Label the Injection to state the content of anhydrous theophylline. USP REFERENCE STANDARDS 11 USP Endotoxin RS USP Theophylline RS
Aminophylline Oral Solution

(Comment on this Monograph)id=m3140=Aminophylline Oral Solution=A-Monos.pdf) DEFINITION Aminophylline Oral Solution is an aqueous solution of Aminophylline, prepared with the aid of Ethylenediamine. It contains an amount of aminophylline equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous theophylline (C7H8N4O2). Aminophylline Oral Solution may contain an excess of ethylenediamine, but no other substance may be added for the purpose of pH adjustment. IDENTIFICATION Sample 1: To an equivalent to 500 mg of aminophylline, add, with constant stirring, 1 mL of 3 N hydrochloric acid or an amount sufficient to precipitate the theophylline completely, and filter (retain the filtrate). Sample 2: Wash the precipitate from Sample 1 with small portions of cold water until free from chloride, and dry at 105 for 1 h. A. PROCEDURE: Sample 2 melts between 270 and 274. B. PROCEDURE Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide to render alkaline, shake by mechanical means for 10 min, add 5 mL of 3 N hydrochloric acid to acidify, chill, collect the precipitated disulfonamide of ethylenediamine, wash with water, recrystallize from water, and dry at 105 for 1 h. Acceptance criteria: Melts between 164 and 171 ASSAY PROCEDURE Mobile phase: 200 mL of methanol, 960 mg of sodium 1pentanesulfonate, and sufficient water to make 1 L. Adjust with glacial acetic acid to a pH of 2.9 0.1. Diluent: Methanol and water (1:4) Standard solution: 0.08 mg/mL of USP Theophylline RS in Diluent Solution A: 0.08 mg/mL of theobromine in the Standard solution System suitability solution: 64 g/mL of theophylline and 64 g/mL of theobromine, from Solution A, in Diluent Sample solution: Equivalent to 72 g/mL of anhydrous theophylline, from Oral Solution in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for theobromine and theophylline are about 0.65 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between theobromine and theophylline, System suitability solution
Aminophylline Rectal Solution

(Comment on this Monograph)id=m3120=Aminophylline Rectal Solution=A-Monos.pdf) DEFINITION Aminophylline Rectal Solution is an aqueous solution of Aminophylline, prepared with the aid of Ethylenediamine. It contains an amount of aminophylline equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous theophylline (C7H8N4O2). Aminophylline Rectal Solution may contain an excess of ethylenediamine, but no other substance may be added for the purpose of pH adjustment. IDENTIFICATION Sample 1: Dilute an equivalent of 500 mg of aminophylline, with water to 20 mL, and add, with constant stirring, 1 mL of 3 N hydrochloric acid or enough to precipitate the theophylline completely, and filter. Sample 2: Wash the precipitate from Sample 1 with a small portion of cold water until free from chloride, and dry at 105 for 4 h. A. INFRARED ABSORPTION 197K Sample: Sample 2 B. PROCEDURE Analysis: To Sample 1, add 0.5 mL of benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide to render alkaline, shake by mechanical means for 10 min, add 5 mL of 3 N hydrochloric acid to acidify, chill, collect the precipitated disulfonamide of ethylenediamine, wash with water, recrystallize from water, and dry at 105 for 1 h.
USP 32
Acceptance criteria: Melts between 164 and 171

Acceptance criteria: Melts between 270 and 274. Analysis 2: To about 10 mg of the dried precipitate from Sample 1, contained in a porcelain dish, add 1 mL of hydrochloric acid and 100 mg of potassium chlorate, evaporate on a steam bath to dryness, and invert the dish over a vessel containing a few drops of 6 N ammonium hydroxide. Acceptance criteria: The residue acquires a purple color, which is destroyed by solutions of fixed alkalies. B. PROCEDURE Sample 2: Filtrate from preparation of Sample 1 (above). Analysis: To Sample 2 add 0.5 mL of benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide to render alkaline, shake by mechanical means for 10 min, add 5 mL of 3 N hydrochloric acid to acidify, chill, collect the precipitated disulfonamide of ethlyenediamine, wash with water, recrystallize from water, and dry at 10 for 1 h. Acceptance criteria: The dried precipitate melts between 164 and 171. ASSAY PROCEDURE Sample composite: Tare a small dish and a glass rod, place in the dish NLT 5 Suppositories, and heat on a steam bath until melted. Mix the melt by stirring it with the rod, cool while stirring, and weigh. Sample stock solution: Weigh a portion of the Sample composite, equivalent to 1 g of aminophylline, place it in a beaker, add 60 mL of hot water and 3 mL of nitric acid, and heat on a steam bath for 15 min with frequent stirring. Cool, transfer to a separator with the aid of 40 mL of ether, shake well, and allow to separate, using a few mL of alcohol, if necessary, to bring about separation of any emulsion that has formed. Draw the water layer into a 100-mL volumetric flask, wash the ether with two 15-mL portions of water, adding the washings to the volumetric flask, dilute with water to volume. Sample solution: Transfer a portion of the Sample stock solution, equivalent to 250 mg of aminophylline, to a 250mL conical flask, add 10 mL of 6 N ammonium hydroxide and 20 mL of 0.1 N silver nitrate VS, and heat on a steam bath for 15 min. Cool to between 5 and 10 for 20 min, then filter, preferably through a filtering crucible of fine porosity under reduced pressure, and wash the precipitate with small portions of water until the last washing gives not more than a faint opalescence with hydrochloric acid. Dissolve the precipitate by pouring over it small volumes of warm 2 N nitric acid, receiving the solution in a conical flask. Wash the filtering crucible a few times with warm water acidified with nitric acid, receiving the washings in the same flask. Cool, and add 2 mL of ferric ammonium sulfate TS. Analysis: Titrate with 0.1 N ammonium thiocyanate VS. Each mL of 0.1 N ammonium thiocyanate is equivalent to 18.02 mg of C7H8N4O2. Acceptance criteria: 90.0%110.0% OTHER COMPONENTS CONTENT OF ETHYLENEDIAMINE Sample: Weigh a portion of the Sample composite from the Assay, equivalent to 500 mg of aminophylline, and place in a 500-mL conical flask. Analysis: Add 150 mL of a mixture of equal volumes of alcohol and ether, and warm gently under reflux for 30 min, with occasional swirling. Cool to room temperature and titrate with 0.1 N hydrochloric acid VS, using a glassmodified calomel electrode system (replace the saturated potassium chloride solution of the calomel electrode with methanol saturated with lithium chloride). Each mL of 0.1 N hydrochloric acid is equivalent to 3.005 mg of ethylenediamine (C2H8N2).
ASSAY PROCEDURE Standard solution: 8 g/mL of USP Theophylline RS in 0.12 M hydrochloric acid Sample stock solution: Equivalent to 1 mg/mL of aminophylline, from Rectal Solution in water Sample solution: 0.01 mg/mL aminophylline from Sample stock solution, in 1.2 M hydrochloric acid, and water (10:89) [NOTEDilute in hydrochloric acid before diluting with water to volume.] Spectrometric conditions Cell: 1 cm Analytical wavelength: Maxima at 270 nm Blank: 0.12 M hydrochloric acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H8N4O2 in each mL of the Rectal Solution taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Theophylline RS in the Standard solution (g/mL) = nominal concentration of anhydrous CU theophylline in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% AU AS CS OTHER COMPONENTS ETHYLENEDIAMINE CONTENT Sample solution: A volume of Rectal Solution equivalent to 500 mg of Aminophylline. Dilute with water, if necessary, to make 30 mL. Analysis: Add methyl orange TS, and titrate with 0.1 N hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is equivalent to 3.005 mg of C2H8N2. Acceptance criteria: 218267 mg of ethylenediamine (C2H8N2) per g of C7H8N4O2 found in the Assay SPECIFIC TESTS PH 791: 9.09.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose or multiple-dose containers, at a controlled room temperature. LABELING: Label the Rectal Solution to state the content of anhydrous theophylline. USP REFERENCE STANDARDS 11 USP Theophylline RS
Aminophylline Suppositories
(Comment on this Monograph)id=m3150=Aminophylline Suppositories=A-Monos.pdf) DEFINITION Aminophylline Suppositories contain an amount of aminophylline equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous theophylline (C7H8N4O2). IDENTIFICATION A. PROCEDURE Sample 1: Evaporate a portion of Sample stock solution from the Assay, equivalent to 500 mg of aminophylline, to about one-half its volume on a steam bath, adjust with 1 N sodium hydroxide to a pH of 7.0, chill, and filter the crystals of theophylline. [NOTESave the filtrate for use in Procedure B.] Analysis 1: Wash crystals from Sample 1 with small portions of ice-cold water, and dry at 105 for 1 h.
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washings with nitric acid, and add an additional 3 mL of the acid. Cool, and add 2 mL of ferric ammonium sulfate TS. Analysis: Titrate the excess silver nitrate with 0.1 N ammonium thiocyanate VS. Each mL of 0.1 N silver nitrate is equivalent to 18.02 mg of C7H8N4O2. Acceptance criteria: 93.0%107.0% OTHER COMPONENTS ETHYLENEDIAMINE CONTENT Sample solution: Transfer the equivalent to 350 mg of aminophylline from powdered Tablets (NLT 20) to a 100-mL conical flask, add 20 mL of water, and digest at 50, with frequent shaking, for 30 min. Cool, filter into a 250-mL conical flask, and wash with water until the last washing is neutral to litmus. To the combined filtrate and washings, add methyl orange TS. Analysis: Titrate with 0.1 N hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is equivalent to 3.005 mg of C2H8N2. Acceptance criteria: 140190 mg of C2H8N2 per g of C7H8N4O2 found in the Assay PERFORMANCE TESTS DISSOLUTION 711 For uncoated or plain coated tablets Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Standard solution: USP Theophylline RS in Medium. Sample solutions: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Detector: UV 269 nm Tolerances: NLT 75% (Q) of the labeled amount of C7H8N4O2 UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Procedure for content uniformity Sample solution: Place 1 Tablet in a 250-mL volumetric flask, add 200 mL of water, and shake by mechanical means until disintegration is complete. Add water to volume. Filter a portion of the mixture, discarding the first 20 mL of the filtrate. Standard solution: 10 g/mL USP Theophylline RS Spectrometric conditions Cell: 1cm Analytical wavelength: 269 nm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the percentage of the label claim of C7H8N4O2 in each Tablet: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Theophylline RS in the Standard solution (g /mL) = concentration of theophylline in the Sample solution (mg/mL)
Acceptance criteria: 152 mg190 mg of C2H8N2 per g of C7H8N4O2 found in the Assay ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, in a cold place. LABELING: Label the Suppositories to state the content of anhydrous theophylline.
Aminophylline Tablets
(Comment on this Monograph)id=m3160=Aminophylline Tablets=A-Monos.pdf) DEFINITION Aminophylline Tablets contain an amount of aminophylline equivalent to NLT 93.0% and NMT 107.0% of the labeled amount of theophylline (C7H8N4O2). [NOTEThe ammoniacal odor present in the vapor space above Aminophylline Tablets is often quite strong, especially when bottles having suitably tight closures are newly opened. This is due to ethylenediamine vapor pressure build-up, a natural condition in the case of aminophylline.] IDENTIFICATION Sample 1: Macerate a quantity of Tablets, equivalent to 500 mg of aminophylline, with 25 mL of water, and filter: the filtrate is alkaline to litmus. To the filtrate, add 1 mL of 3 N hydrochloric acid, stir, and chill, if necessary, to precipitate the theophylline. Filter, and retain the filtrate, free from washings. Sample 2: Wash the crystals obtained from Sample 1 on the filter with small quantities of ice-cold water, and dry at 105 for 1 h. A. To 10 mg of Sample 2, in a porcelain dish, add 1 mL of hydrochloric acid and 100 mg of potassium chlorate, evaporate on a steam bath to dryness, and invert the dish over a vessel containing a few drops of 6 N ammonium hydroxide: the residue acquires a purple color, which is destroyed by solutions of fixed alkalies. B. Recrystallize Sample 2 from water and dry at 105 for 1 h: it melts between 270 and 274. C. To about 25 mL of Sample 1, add 0.5 mL of benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide to render alkaline. Shake by mechanical means for 10 min. Add 5 mL of 3 N hydrochloric acid to acidify. Chill, collect the precipitated disulfonamide of ethylenediamine, wash with water, recrystallize from water, and dry at 105 for 1 h: the dried precipitate melts between 164 and 171. ASSAY PROCEDURE Sample solution: Transfer the equivalent to 2 g of aminophylline from powdered Tablets (NLT 20) to a 200-mL volumetric flask, and add 50 mL of water and 15 mL of 6 N ammonium hydroxide. Allow to stand for 30 min with frequent shaking, warming to 50, if necessary, to dissolve the aminophylline. Cool the mixture to room temperature if it has been warmed, and add water to volume. Centrifuge 50 mL of the mixture, and pipet a portion of the clear supernatant, equivalent to 250 mg of aminophylline, into a 250-mL conical flask, and dilute with water, if necessary, to make 40 mL. Add 8 mL of 6 N ammonium hydroxide and 20.0 mL of 0.1 N silver nitrate VS, mix, heat to boiling, and continue boiling for 15 min. Cool to between 5 and 10 for 20 min, then filter, preferably through a filtering crucible under reduced pressure, and wash the precipitate with three 10-mL portions of water. Acidify the combined filtrate and
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Tablets to state the content of anhydrous theophylline. USP REFERENCE STANDARDS 11 USP Theophylline RS
USP 32

means until disintegration is complete. Add water to volume. Filter a portion of the mixture, discarding the first 20 mL of the filtrate. Spectrometric conditions Cell: 1 cm Analytical wavelength: 269 nm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the label claim percentage of C7H8N4O2 in each Tablet: Result = (AU/AS) (CS /CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Theophylline RS in the Standard solution (g/mL) = concentration of theophylline in the Sample solution (g/mL)
Aminophylline Delayed-Release Tablets

(Comment on this Monograph)id=m3165=Aminophylline Delayed-Release Tablets=A-Monos.pdf) DEFINITION Aminophylline Delayed-Release Tablets contain an amount of aminophylline equivalent to NLT 93.0% and NMT 107.0% of the labeled amount of anhydrous theophylline (C7H8N4O2). [NOTEThe ammoniacal odor present in the vapor space above Aminophylline Delayed-Release Tablets is often quite strong, especially when bottles having suitably tight closures are newly opened. This is due to ethylenediamine vapor pressure buildup, a natural condition in the case of aminophylline.] ASSAY PROCEDURE Sample solution: Transfer an equivalent to 2 g of aminophylline, from powdered Tablets (NLT 20), to a 200mL volumetric flask. Add 50 mL of water and 15 mL of 6 N ammonium hydroxide, and allow to stand for 30 min with frequent shaking, warming to 50, if necessary, to dissolve the aminophylline. Cool the mixture to room temperature if it has been warmed, and add water to volume. Centrifuge 50 mL of the mixture, and pipet a portion of the clear supernatant, equivalent to 250 mg of aminophylline, into a 250-mL conical flask, and dilute with water, if necessary, to make 40 mL. Add 8 mL of 6 N ammonium hydroxide and 20.0 mL of 0.1 N silver nitrate VS, mix, heat to boiling, and continue boiling for 15 min. Cool to between 5 and 10 for 20 min, then filter, preferably through a filtering crucible under reduced pressure, and wash the precipitate with three 10-mL portions of water. Acidify the combined filtrate and washings with nitric acid, and add an additional 3 mL of the acid. Cool, and add 2 mL of ferric ammonium sulfate TS. Analysis: Titrate the excess silver nitrate with 0.1 N ammonium thiocyanate VS. Each mL of 0.1 N silver nitrate is equivalent to 18.02 mg of C7H8N4O2. Acceptance criteria: 93.0%107.0% OTHER COMPONENTS ETHYLENEDIAMINE CONTENT Sample solution: Weigh a portion of the powdered Tablets prepared in the Assay, equivalent to 350 mg of aminophylline, transfer to a 100-mL conical flask, add 20 mL of water, and digest at 50, with frequent shaking, for 30 min. Cool, filter into a 250-mL conical flask, and wash with water until the last washing is neutral to litmus. To the combined filtrate and washings add methyl orange TS. Analysis: Titrate with 0.1 N hydrochloric acid VS. Each mL of 0.1 N hydrochloric acid is equivalent to 3.005 mg of C2H8N2. Acceptance criteria: 140190 mg of C2H8N2 per g of C7H8N4O2 found in the Assay PERFORMANCE TESTS DISINTEGRATION, Procedure 701: 30 min, determined as directed under Delayed-Release (Enteric-Coated) Tablets UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Standard solution: 10 g/mL USP Theophylline RS Sample solution: Place 1 Tablet in a 250-mL volumetric flask, add 200 mL of water, and shake by mechanical
SPECIFIC TESTS OTHER REQUIREMENTS Sample 1: Macerate a quantity of Tablets, equivalent to 500 mg of aminophylline, with 25 mL of water, and filter: the filtrate is alkaline to litmus. To the filtrate add 1 mL of 3 N hydrochloric acid, stir, and chill, if necessary, to precipitate the theophylline. Filter, and retain the filtrate, free from washings. Sample 2: Wash the crystals obtained from Sample 1 on the filter with small quantities of ice-cold water, and dry at 105 for 1 h. Procedure 1 Analysis: To 10 mg of Sample 2, in a porcelain dish, add 1 mL of hydrochloric acid and 100 mg of potassium chlorate, evaporate on a steam bath to dryness, and invert the dish over a vessel containing a few drops of 6 N ammonium hydroxide. Acceptance criteria: The residue acquires a purple color, which is destroyed by solutions of fixed alkalies. Procedure 2 Analysis: Recrystallize Sample 2 from water and dry at 105 for 1 h. Acceptance criteria: It melts between 270 and 274. Procedure 3 Analysis: To about 25 mL of Sample 1, add 0.5 mL of benzenesulfonyl chloride and 5 mL of 1 N sodium hydroxide to render alkaline, and shake by mechanical means for 10 min. Add 5 mL of 3 N hydrochloric acid to acidify, chill, collect the precipitated disulfonamide of ethylenediamine, wash with water, recrystallize from water, and dry at 105 for 1 h. Acceptance criteria: The precipitate melts between 164 and 171. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Tablets to state the content of anhydrous theophylline. USP REFERENCE STANDARDS 11 USP Theophylline RS
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Suitability requirements Resolution: NLT 1.7 between aminosalicylic acid and acetaminophen Relative standard deviation: NMT 1.0% for the peak response ratio of the aminosalicylic acid peak to the acetaminophen peak Analysis Samples: Standard solution and Sample solution [NOTEAfter use, wash the column for 30 min with methanol, water, and phosphoric acid (77:23:0.6), and then wash for 30 min with methanol and water (50:50).] Calculate the percentage of C7H6NNaO3 taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratio of aminosalicylate peak to the acetaminophen peak from the Sample solution = peak response ratio of aminosalicylic acid peak RS to the acetaminophen peak from the Standard solution = concentration of USP Aminosalicylic Acid RS in CS the Standard solution (mg/mL) = concentration of aminosalicylate in the Sample CU solution (mg/mL) = molecular weight of anhydrous aminosalicylate Mr1 sodium, 175.12 Mr2 = molecular weight of aminosalicylic acid, 153.14 Acceptance criteria: 98.0%101.0% RU IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221 Sample solution: 25 mg/mL in 4 M nitric acid Acceptance criteria: The solution shows no more chloride than corresponds to 0.30 mL of 0.020 N hydrochloric acid (0.042%). HEAVY METALS, Method II 231: NMT 30 ppm Organic Impurities PROCEDURE: LIMIT OF M-AMINOPHENOL Solution A: 12.7 mg/mL tetrabutylammonium hydroxide in methanol Mobile phase: Solution A, 0.05 M dibasic sodium phosphate, and 0.05 M monobasic sodium phosphate (30:85:85) Internal standard solution: 5 g/mL of sulfanilamide in Mobile phase Standard stock solution: 12 g/mL of USP mAminophenol RS in Mobile phase Standard solution: Standard stock solution, Internal standard solution, and Mobile phase (1:1:8) in a low-actinic volumetric flask Sample solution: Use the Sample solution, prepared as directed in the Assay. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; 10-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for sulfanilamide and m-aminophenol are 0.66 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between m-aminophenol and sulfanilamide
Aminosalicylate Sodium
(Comment on this Monograph)id=m3340=Aminosalicylate Sodium=A-Monos.pdf) C7H6NNaO3 2H2O 211.15 Benzoic acid, 4-amino-2-hydroxy-, monosodium salt, dihydrate; Monosodium 4-aminosalicylate dihydrate [6018-19-5]. Anhydrous [133-10-8]. 175.12 DEFINITION Aminosalicylate Sodium contains NLT 98.0% and NMT 101.0% of C7H6NNaO3, calculated on the anhydrous basis. [CAUTIONPrepare solutions of Aminosalicylate Sodium within 24 h of administration. Under no circumstances use a solution if its color is darker than that of a freshly prepared solution.] IDENTIFICATION A. Dissolve 250 mg in 3 mL of 1 N sodium hydroxide, transfer to a 500-mL volumetric flask, and dilute with water to volume. Transfer a 5-mL aliquot to a 250-mL volumetric flask containing 12.5 mL of pH 7 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions), and dilute with water to volume. This solution, when compared in a suitable spectrophotometer against a blank of the same buffer in the same concentration, exhibits absorbance maxima at 265 2 nm and 299 2 nm, and the ratio A265/A299 is between 1.50 and 1.56. B. PROCEDURE Analysis: Place 1-g sample in a small, round-bottom flask, and add 10 mL of acetic anhydride. Heat the flask on a steam bath for 30 min, add 40 mL of water, filter, cool, and allow to stand until the diacetyl derivative has crystallized. Collect the precipitate on a filter, wash well with water, and dry at 105 for 1 h. Acceptance criteria: The diacetyl derivative melts between 191 and 197. C. PROCEDURE: Dissolve 50 mg in 5 mL of water, add 1 mL of 3 N hydrochloric acid, and filter if necessary. To the filtrate add 1 drop of ferric chloride TS: a violet color is produced. D. IDENTIFICATIONS TESTSGENERAL, Sodium 191: Meets the requirements ASSAY PROCEDURE Solution A: 12.7 mg/mL tetrabutylammonium hydroxide in methanol Mobile phase: Solution A, 0.05 M dibasic sodium phosphate, and 0.05 M monobasic sodium phosphate (30:85:85) Internal standard solution: 5 mg/mL of acetaminophen in Mobile phase Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid RS and 0.5 mg/mL of acetaminophen from Internal standard solution in Mobile phase [NOTEDissolve in a portion of Mobile phase before diluting to final volume. Use a lowactinic volumetric flask.] Sample solution: 0.69 mg/mL of Aminosalicylate Sodium in Mobile phase and Internal standard solution (9:1) [NOTE Dissolve in a portion of Mobile phase before diluting to final volume. Use low-actinic glass ware.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen and aminosalicylic acid are 0.83 and 1.0, respectively.]
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Relative standard deviation: NMT 7% Analysis Samples: Standard solution and Sample solution [NOTEAfter use, wash the column for 30 min with methanol, water, and phosphoric acid (77:23:0.6), and then wash for 30 min with methanol and water (50:50).] Calculate the percentage of m-aminophenol, in the portion of Aminosalicylate Sodium taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the m-aminophenol peak to the sulfanilamide peak from the Sample solution = peak response ratio of the m-aminophenol peak RS to the sulfanilamide peak from the Standard solution = concentration of USP Aminophenol RS in the CS Standard solution (g/mL) = concentration for the Sample solution (mg/mL) CU Acceptance criteria: NMT 0.25% SPECIFIC TESTS PH 791 Sample solution: 20 mg/mL Acceptance criteria: 6.58.5 WATER DETERMINATION, Method I 921: Sample solution: 20 mg/ml Acceptance criteria: 16.0%18.0% HYDROGEN SULFIDE, SULFUR DIOXIDE, AND AMYL ALCOHOL: Dissolve 500 mg in 5 mL of 1 N sodium hydroxide, add 6 mL of 3 N hydrochloric acid, and stir vigorously: no odor of hydrogen sulfide or sulfur dioxide is perceptible, and not more than a faint odor of amyl alcohols is perceptible. A piece of moistened lead acetate test paper held over the mixture does not become discolored. CLARITY AND COLOR OF SOLUTION: One g dissolves in 10 mL of water to give a clear solution that has not more than a faint yellow color. One g dissolves in a freshly prepared mixture of 5 mL of nitric acid and 45 mL of water to give a clear solution that has not more than a slight color. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, protected from excessive heat. USP REFERENCE STANDARDS 11 USP Aminosalicylic Acid RS USP m-Aminophenol RS RU
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allow to stand until the diacetyl derivative has crystallized. Collect the precipitate on a filter, wash well with water, and dry at 105 for 1 h. Acceptance criteria: The diacetyl derivative melts between 191 and 197. B. PROCEDURE Analysis: Shake 0.1 g of Sample with 10 mL of water, and filter. To 5 mL of the filtrate, add 1 drop of ferric chloride TS. Acceptance criteria: A violet color is produced. ASSAY PROCEDURE Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide in methanol Mobile phase: Solution A, 0.05 M dibasic sodium phosphate, and 0.05 M monobasic sodium phosphate (30:85:85) Internal standard solution: 5 mg/mL of acetaminophen in Mobile phase Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid RS and 0.5 mg/mL of acetaminophen from the Internal standard solution in Mobile phase and the Internal standard solution (9:1) [NOTEDissolve in a portion of Mobile phase before diluting to final volume. Use a low-actinic volumetric flask.] Sample stock solution: Add the equivalent of 690 mg of aminosalicylate sodium from the powdered Tablets to a 100mL low-actinic volumetric flask. Add 50 mL of Mobile phase, and shake for 5 min. Dilute with Mobile phase to volume, and filter. Sample solution: 0.69 mg/mL of aminosalicylic acid from the Sample stock solution and 0.5 mg/mL of acetaminophen from the Internal standard solution in Mobile phase [NOTE Use a low-actinic volumetric flask.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen and aminosalicylic acid are 0.83 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.7 between aminosalicylic acid and acetaminophen Relative standard deviation: NMT 1.0%, ratios of the peak response of aminosalicylic acid to the peak response of acetaminophen Analysis Sample: Sample solution and Standard solution [NOTEAfter use, wash the column for 30 min with a mixture of methanol, water, and phosphoric acid (77:23:0.6), and then wash for 30 min with a mixture of methanol and water (50:50).] Calculate the percentage of C7H6NNaO3 2H2O in the portion of Tablets taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) (100/L) RU RS CS = peak response ratio of aminosalicylate to acetaminophen from the Sample solution = peak response ratio of aminosalicylic acid to acetaminophen from the Standard solution = concentration of USP Aminosalicylic Acid RS in the Standard solution (mg/mL)
Aminosalicylate Sodium Tablets

(Comment on this Monograph)id=m3370=Aminosalicylate Sodium Tablets=A-Monos.pdf) DEFINITION Aminosalicylate Sodium Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of C7H6NNaO3 2H2O. IDENTIFICATION Sample: Mix powdered Tablets, equivalent to 3 g of aminosalicylate sodium, with 40 mL of water, and filter. Add to the filtrate 15 mL of 1 N acetic acid, and allow to stand until precipitation has occurred. Collect the precipitate on a filter, wash well with water, and dry at 105 for 30 min. A. PROCEDURE Analysis: Place 1 g of Sample in a small, round-bottom flask, and add 10 mL of acetic anhydride. Heat the flask on a steam bath for 30 min, add 40 mL of water, filter, cool, and
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Aminosalicylate / Official Monographs

RS
USP 32
= peak response ratio of m-aminophenol to sulfanilamide from the Standard solution = concentration of USP m-Aminophenol RS in the CS Standard solution (g/mL) = concentration of aminosalicylate sodium in the CU Sample solution, as determined in the Assay (mg/mL) F = conversion factor (g/mg) Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, protected from excessive heat. USP REFERENCE STANDARDS 11 USP Aminosalicylic Acid RS USP m-Aminophenol RS
= concentration of aminosalicylate sodium in the Sample solution (unit/mL) = molecular weight of aminosalicylate sodium Mr1 dihydrate, 211.15 = molecular weight of aminosalicylic acid, 153.14 Mr2 L = label claim (mg/unit) Acceptance criteria: 95.0%105.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Analysis: Determine the amount of C7H6NNaO3 2H2O dissolved, employing the procedure set forth in the Assay, making any necessary modifications. Tolerances: NLT 75% (Q) of C7H6NNaO3 2H2O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities LIMIT OF m-AMINOPHENOL Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide in methanol Mobile phase: Solution A, 0.05 M dibasic sodium phosphate, and 0.05 M monobasic sodium phosphate (30:85:85) Internal standard solution: 5 g/mL of sulfanilamide in Mobile phase Standard stock solution: 12 g/mL of USP mAminophenol RS in Mobile phase Standard solution: Standard stock solution, Internal standard solution, and Mobile phase (1:1:8) in a low-actinic volumetric flask Sample stock solution: Add the equivalent of 690 mg of aminosalicylate sodium from the powdered Tablets, to a 100-mL low-actinic volumetric flask. Add 50 mL of Mobile phase, and shake for 5 min. Dilute with Mobile phase to volume, and filter. Sample solution: 0.69 mg/mL of aminosalicylic acid from the Sample stock solution and 0.5 mg/mL of acetaminophen from the Internal standard solution in Mobile phase [NOTE Use a low-actinic volumetric flask.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; 10-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for sulfanilamide and m-aminophenol are 0.66 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between m-aminophenol and sulfanilamide Relative standard deviation: NMT 7% Analysis Sample: Sample solution and Standard solution [NOTEAfter use, wash the column for 30 min with a mixture of methanol, water, and phosphoric acid (77:23:0.6), and then wash for 30 min with a mixture of methanol and water (50:50).] Calculate the percentage of m-aminophenol in the portion of Tablets taken: Result = (RU/RS) (CS/CU) F 100 RU = peak response ratio of m-aminophenol to sulfanilamide from the Sample solution
Aminosalicylic Acid
(Comment on this Monograph)id=m3400=Aminosalicylic Acid=A-Monos.pdf)
C7H7NO3 Benzoic acid, 4-amino-2-hydroxy-; 4-Aminosalicylic acid [65-49-6].
153.14
DEFINITION Aminosalicylic Acid contains NLT 98.5% and NMT 100.5% of C7H7NO3, calculated on the anhydrous basis. [CAUTIONUnder no circumstances use a solution prepared from Aminosalicylic Acid if its color is darker than that of a freshly prepared solution.] IDENTIFICATION A. Procedure Sample solution: Dissolve 250 mg in 3 mL of 1 N sodium hydroxide, transfer to a 500-mL volumetric flask, and dilute with water to volume. Analysis: Transfer a 5-mL aliquot of the Sample solution to a 250-mL volumetric flask containing 12.5 mL of pH 7 phosphate buffer (see Reagents, Indicators, and Solutions Buffer Solutions), and dilute with water to volume. This solution, when compared in a suitable spectrophotometer against a blank of the same buffer in the same concentration, exhibits absorbance maxima at 265 2 and 299 2 nm, and the ratio A265/A299 is between 1.50 and 1.56. B. Place 1 g in a small, round-bottom flask, and add 10 mL of acetic anhydride. Heat the flask on a steam bath for 30 min, add 40 mL of water, filter, cool, and allow to stand until the diacetyl derivative has crystallized. Collect the precipitate on a filter, wash well with water, and dry at 105 for 1 h. Acceptance criteria: The diacetyl derivative so obtained melts between 191 and 197. C. Shake 0.1 g with 10 mL of water, and filter. To 5 mL of the filtrate, add 1 drop of ferric chloride TS. Acceptance criteria: A violet color is produced. ASSAY PROCEDURE Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide in methanol
USP 32
Mobile phase: Solution A, 0.05 M dibasic sodium phosphate, and 0.05 M monobasic sodium phosphate (30:85:85) Internal standard solution: 5 mg/mL of acetaminophen in Mobile phase Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid RS and 0.5 mg/mL of acetaminophen from Internal standard solution in a mixture of Mobile phase and Internal standard solution (9:1) [NOTEDissolve in a portion of Mobile phase before diluting to final volume. Use a low-actinic volumetric flask.] Sample solution: 0.5 mg/mL of Aminosalicylic Acid in a mixture of Mobile phase and Internal standard solution (9:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen and for aminosalicylic acid are 0.83 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.7 between aminosalicylic acid and acetaminophen Relative standard deviation: NMT 1.0% for the peak response ratios of the aminosalicylic acid to the acetaminophen peaks Analysis Sample: Sample solution and Standard solution [NOTEAfter use, wash the column for 30 min with a mixture of methanol, water, and phosphoric acid (77:23:0.6), and then wash for 30 min with a mixture of methanol and water (50:50).] Calculate the percentage of C7H7NO3 in the portion of Aminosalicylic Acid taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of aminosalicylic acid to acetaminophen from the Sample solution = peak response ratio of aminosalicylic acid to RS acetaminophen from the Standard solution = concentration of USP Aminosalicylic acid RS in CS the Standard solution (mg/mL) CU = concentration of aminosalicylic acid in the Sample solution (mg/mL) Acceptance criteria: 98.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% CHLORIDE AND SULFATE, Chloride 221 Sample solution: 25 mg/mL in 4 M nitric acid Acceptance criteria: The solution shows no more chloride than corresponds to 0.30 mL of 0.020 N hydrochloric acid (0.042%). HEAVY METALS, Method II 231: NMT 30 ppm Organic Impurities LIMIT OF m-AMINOPHENOL Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide in methanol Mobile phase: Solution A, 0.05 M dibasic sodium phosphate, and 0.05 M monobasic sodium phosphate (150:425:425) Internal standard solution: 5 g/mL of sulfanilamide in Mobile phase RU
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Standard stock solution: 12 g/mL of USP mAminophenol RS in Mobile phase Standard solution: Standard stock solution, Internal standard solution, and Mobile phase (1:1:8) in a low-actinic volumetric flask Sample solution: 0.5 mg/mL of Aminosalicylic Acid in a mixture of Mobile phase and Internal standard solution (9:1) [NOTEDissolve in a portion of Mobile phase before diluting to final volume. Use a low-actinic volumetric flask.] Chromatographic system: (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; 10-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTERelative retention times for sulfanilamide and for m-aminophenol are about 0.66 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between m-aminophenol and sulfanilamide Relative standard deviation: NMT 7% Analysis Sample: Sample solution and Standard solution [NOTEAfter use, wash the column for 30 min with a mixture of methanol, water, and phosphoric acid (77:23:0.6), and then wash for 30 min with a mixture of methanol and water (50:50).] Calculate the percentage of m-aminophenol, in the portion of Aminosalicylic Acid taken: Result = (RU/RS) (CS/CU) F 100 = peak response ratio of m-aminophenol to sulfanilamide from the Sample solution = peak response ratio of the m-aminophenol to RS sulfanilamide from the Standard solution = concentration of USP m-Aminophenol RS in the CS Standard solution (g/mL) CU = concentration of the Sample solution (mg/mL) F = conversion factor (g/mg) Acceptance criteria: NMT 0.25% RU SPECIFIC TESTS PH 791: 3.03.7, in a saturated solution WATER DETERMINATION, Method I 921: NMT 0.5% HYDROGEN SULFIDE, SULFUR DIOXIDE, AND AMYL ALCOHOL Sample solution: Dissolve 500 mg in 5 mL of 1 N sodium hydroxide, add 6 mL of 3 N hydrochloric acid, and stir vigorously. Acceptance criteria: No odor of hydrogen sulfide or sulfur dioxide is perceptible, and NMT a faint odor of amyl alcohol is perceptible. A piece of moistened lead acetate test paper held over the mixture does not become discolored. CLARITY AND COLOR OF SOLUTION: One g dissolves in 10 mL of sodium bicarbonate solution (1 in 15) to form a clear solution that has NMT a faint yellow color. One g dissolves in 50 mL of freshly prepared 1.6 M nitric acid to form a clear solution that has NMT a slight color. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at a temperature not exceeding 30. USP REFERENCE STANDARDS 11 USP Aminosalicylic Acid RS USP m-Aminophenol RS
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(77:23:0.6), and then wash for 30 min with a mixture of methanol and water (50:50).] Calculate the percentage of C7H7NO3 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of aminosalicylic acid to acetaminophen from the Sample solution = peak response ratio of aminosalicylic acid to RS acetaminophen from the Standard solution = concentration of USP Aminosalicylic Acid RS in CS the Standard solution (mg/mL) = concentration of aminosalicylic acid in the CU Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: pH 7.5 phosphate buffer; 900 mL (see Reagents, Indicators, and SolutionsBuffer Solutions) Apparatus 1: 100 rpm Time: 45 min Analysis: Determine the amount of C7H7NO3 dissolved, employing the procedure in the Assay, making any necessary modifications. Tolerances: NLT 75% (Q) of C7H7NO3 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF m-AMINOPHENOL Mobile phase and Internal standard solution: Prepare as directed in the Assay. Standard stock solution: 12 g/mL of USP mAminophenol RS in Mobile phase Standard solution: Standard stock solution, Internal standard solution, and Mobile phase (1:1:8) in a low-actinic volumetric flask Sample solution: Use the Sample solution in the Assay. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; 10-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for sulfanilamide and for m-aminophenol are 0.66 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between m-aminophenol and sulfanilamide Relative standard deviation: NMT 7% Analysis Sample: Sample solution and Standard solution [NOTEAfter use, wash the column for 30 min with a mixture of methanol, water, and phosphoric acid (77:23:0.6), and then wash for 30 min with a mixture of methanol and water (50:50).] Calculate the percentage of m-aminophenol, in the portion of Tablets taken: Result = (RU/RS) (CS/CU) F 100 RU RS CS = peak response ratio of m-aminophenol to sulfanilamide from the Sample solution = peak response ratio of m-aminophenol to sulfanilamide from the Standard solution = concentration of USP m-Aminophenol RS in the Standard solution (g/mL) RU
Aminosalicylic Acid Tablets

(Comment on this Monograph)id=m3430=Aminosalicylic Acid Tablets=A-Monos.pdf) DEFINITION Aminosalicylic Acid Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of C7H7NO3. IDENTIFICATION Sample: Mix powdered Tablets, equivalent to 2 g of aminosalicylic acid, with 50 mL of a mixture of acetone and chloroform (1:2), and filter. Evaporate the filtrate with a current of warm air to dryness. A. PROCEDURE Analysis: Place 1 g of Sample in a small, round-bottom flask, and add 10 mL of acetic anhydride. Heat the flask on a steam bath for 30 min, add 40 mL of water, filter, cool, and allow to stand until the diacetyl derivative has crystallized. Collect the precipitate on a filter, wash well with water, and dry at 105 for 1 h. Acceptance criteria: The diacetyl derivative melts between 191 and 197. B. PROCEDURE Analysis: Shake 0.1 g of Sample with 10 mL of water, and filter. To 5 mL of the filtrate, add 1 drop of ferric chloride TS. Acceptance criteria: A violet color is produced. ASSAY PROCEDURE Solution A: 12.7 mg/mL of tetrabutylammonium hydroxide in methanol Mobile phase: Solution A, 0.05 M dibasic sodium phosphate, and 0.05 M monobasic sodium phosphate (30:85:85) Internal standard solution: 5 mg/mL of acetaminophen in Mobile phase Standard solution: 0.5 mg/mL of USP Aminosalicylic Acid RS and 0.5 mg/mL of acetaminophen from the Internal standard solution in Mobile phase [NOTEDissolve in a portion of Mobile phase before diluting to final volume. Use a low-actinic volumetric flask.] Sample stock solution: Add the equivalent of 500 mg of aminosalicylic acid from the powdered Tablets, to a 100-mL low-actinic volumetric flask. Add 50 mL of Mobile phase, and shake for 5 min. Dilute with Mobile phase to volume, and filter. Sample solution: 0.5 mg/mL of aminosalicylic acid from the Sample stock solution and 0.5 mg/mL of acetaminophen from the Internal standard solution in Mobile phase [NOTE Use a low-actinic volumetric flask.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen and for aminosalicylic acid are 0.83 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.7 between aminosalicylic acid and acetaminophen Relative standard deviation: NMT 1.0%, ratios of the peak response of aminosalicylic acid to the peak response of acetaminophen Analysis Sample: Sample solution and Standard solution [NOTEAfter use, wash the column for 30 min with a mixture of methanol, water, and phosphoric acid
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= concentration of aminosalicylic acid in the portion of Tablets taken for the Sample solution as determined in the Assay (mg/mL) F = conversion factor (g/mg) Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at a temperature not exceeding 30. USP REFERENCE STANDARDS 11 USP Aminosalicylic Acid RS USP m-Aminophenol RS CU
Official Monographs / Amitraz 205

Temperature Column: 250 Detector: 250 Carrier gas: Dry nitrogen Flow rate: 60 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 3.0 between squalane and amitraz Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution, Standard solution, and System suitability solution Calculate the percentage of C19H23N3 in the portion of Amitraz taken: Result = (RU/RS) (CS/CU) 100 = ratio of the responses of the amitraz and squalane peaks from the Sample solution = ratio of the responses of the amitraz and RS squalane peaks from the Standard solution = concentration of USP Amitraz RS in the Standard CS solution (mg/mL) = concentration of amitraz in the Sample solution CU (mg/mL) Acceptance criteria: 95.0%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Standard solution A: 2.0 mg/mL of USP Amitraz RS in toluene Standard solution B: 0.30 mg/mL of 2,4-dimethylaniline in toluene Sample solution: 100 mg/mL of Amitraz in toluene Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2 L Developing solvent system: Cyclohexane, ethyl acetate, and triethylamine (5:3:2) Spray reagent: 0.5% solution of N-(1-naphthyl) ethylenediamine dihydrochloride in methanol Analysis Samples: Sample solution, Standard solution A, and Standard solution B Stand the plate to a depth of 3.5 cm in a solution prepared by dissolving 35 g of acetamide in 100 mL of methanol, adding 100 mL of triethylamine, and diluting to 250 mL with methanol. Allow the wet plate to stand in a current of cold air for 30 s. Immediately apply Samples separately to the plate, at a level about 1 cm below the top of the impregnated zone. Promptly develop the plate. Examine the plate under short-wavelength UV light. Acceptance criteria 1: Any secondary spot from the Sample solution is not more intense than the corresponding spot from the Standard solution A (2.0%). Expose the plate to the vapor of hydrochloric acid for 10 min, then expose it to the vapor of nitrogen dioxide RU
Amitraz
(Comment on this Monograph)id=m3465=Amitraz=AMonos.pdf)
293.41 C19H23N3 Methanimidamide, N-(2,4-dimethylphenyl)-[[N-(2,4dimethylphenyl)imino]methyl-N]-methyl-; N-Methyl-N-2,4-xylyl-N-(N-2,4-xylylformimidoyl)formamidine; N-Methylbis(2,4-xylyliminomethyl)amine [33089-61-1]. DEFINITION Amitraz contains NLT 95.0% and NMT 101.5% of C19H23N, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Sample solution: 2 mg/mL of Amitraz in toluene Standard solution: 2 mg/mL of USP Amitraz RS in toluene Analysis: Proceed as directed for Organic Impurities, Procedure. Acceptance criteria: The RF value of the principal spot in the Sample solution corresponds to that of the Standard solution. C. The retention time of the major peak from the System suitability solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Internal standard solution: 10 mg/mL of squalane in methyl acetate Standard solution: 8 mg/mL of USP Amitraz RS in Internal standard solution Sample solution: 8 mg/mL of Amitraz in Internal standard solution System suitability solution: 8 mg/mL of Amitraz in methyl acetate Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 1.5-m column packed with 3% liquid phase G1 on support S1A
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ASSAY PROCEDURE Internal standard solution: 10 mg/mL of squalane in methyl acetate Standard solution: 8 mg/mL of USP Amitraz RS in Internal standard solution Sample solution: Equivalent to 8 mg/mL of Amitraz, from the volume of Concentrate in the Internal standard solution System suitability solution: Equivalent to 8 mg/mL of Amitraz, from the volume of the Concentrate in methyl acetate Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 1.5-m column packed with 3% liquid phase G1 on support S1A Temperature Column: 250 Detector: 250 Carrier gas: Dry nitrogen Flow rate: 60 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 3.0 between squalane and amitraz Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution, Standard solution, and System suitability solution Calculate the percentage of C19H23N3 in the Concentrate taken: Result = (RU/RS) (CS/CU) 100 = ratio of the response of the amitraz peak to the response of the squalane peak from the Sample solution RS = ratio of the response of the amitraz peak to the response of the squalane peak from the Standard solution CS = concentration of USP Amitraz RS in the Standard solution (mg/mL) CU = nominal concentration of amitraz in the Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% RU SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.15%, with anhydrous pyridine being used in place of methanol in the titration vessel ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only, and that it is to be diluted before use. The label states the name and quantity of diluent to be used, the directions for dilution, and the conditions for storage of the constituted Dip. USP REFERENCE STANDARDS 11 USP Amitraz RS
(prepared by the reaction of nitric acid and zinc) for 10 min, remove any excess nitric oxide by air exhaust, spray the plate with Spray reagent, and examine the plate. Acceptance criteria 2: Any secondary spot from the Sample solution corresponding to 2,4-dimethylaniline is not more intense than the spot from the Standard solution B (0.30%). SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.1%, anhydrous pyridine being used in place of methanol in the titration vessel ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Amitraz RS
Amitraz Concentrate for Dip

(Comment on this Monograph)id=m3470=Amitraz Concentrate for Dip=A-Monos.pdf) DEFINITION Amitraz Concentrate for Dip contains Amitraz in a suitable vehicle. It may contain a suitable stabilizing agent. It contains NLT 90.0% and NMT 120.0% of the labeled amount of C19H23N3. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 5 mg/mL of USP Amitraz RS in toluene Sample solution: 5 mg/mL of Amitraz from Concentrate for Dip diluted with toluene Application volume: 2 L Developing solvent system: Cyclohexane, ethyl acetate, and triethylamine (5:3:2) Spray reagent: 0.5% solution of N-(1-naphthyl) ethylenediamine dihydrochloride in methanol Analysis Samples: Sample solution and Standard solution Pre-treatment: Stand the plate to a depth of 3.5 cm in a solution prepared by dissolving 35 g of acetamide in 100 mL of methanol, adding 100 mL of triethylamine, and diluting to 250 mL with methanol. Allow the wet plate to stand in a current of cold air for 30 s. Immediately apply Samples separately to the plate, at a level about 1 cm below the top of the impregnated zone. Promptly develop the plate. Proceed as directed in Chromatography 621, Thin-Layer Chromatography. Examine the plate under shortwavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. The retention time of the major peak of the System suitability solution corresponds to that of the Standard solution, as obtained in the Assay.
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Official Monographs / Amitriptyline 207

Mode: LC Detector: UV 215 nm Column: 4.6-mm 25-cm; 5-m packing L7 Temperature: 45 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEFor relative retention times, see Impurity Table 1.] Suitability requirements Resolution: NLT 1.5 between amitriptyline related compound B and nortriptyline, System suitability solution Relative standard deviation: NMT 2.0% for amitriptyline, Standard solution Analysis Samples: Standard solution and Sample solution [NOTERecord the chromatograms for 40 min.] Calculate the percentage of C20H23N HCl in the portion of Amitriptyline Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amitriptyline Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of amitriptyline hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Diluted phosphoric acid, Buffer, Mobile phase, Chromatographic system, and System suitability: Proceed as directed in the Assay. Standard solution: Use the System suitability solution, prepared as directed in the Assay. Sample solution: Use the Sample stock solution, prepared as directed in the Assay. Analysis Samples: Sample solution and Standard solution Calculate the percentages of individual amitriptyline related compounds in the portion of Amitriptyline Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = individual peak response for each amitriptyline related compound of the Sample solution = response of the corresponding peak in the Standard solution = concentration of each of the amitriptyline related compounds in the Standard solution (mg/mL) rU rS CS
Amitriptyline Hydrochloride
(Comment on this Monograph)id=m3480=Amitriptyline Hydrochloride=A-Monos.pdf)
313.86 C20H23N HCl 1-Propanamine, 3-(10,11-dihydro-5H-dibenzo [a,d]cyclohepten-5-ylidene)-N,N-dimethyl-, hydrochloride; 10,11-Dihydro-N,N-dimethyl-5H-dibenzo[a,d]cycloheptene-5,propylamine hydrochloride [549-18-8]. DEFINITION Amitriptyline Hydrochloride contains NLT 98.0% and NMT 102.0% of C20H23N HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Diluted phosphoric acid: Phosphoric acid and water (1:10) Buffer: Dissolve 1.42 mg/mL of dibasic sodium phosphate (Na2HPO4) in water, and adjust with diluted phosphoric acid to a pH of 7.7. Mobile phase: Methanol and Buffer (7:3) Standard solution: 0.2 mg/mL of USP Amitriptyline Hydrochloride RS in Mobile phase Sample stock solution: 1 mg/mL of Amitriptyline Hydrochloride in Mobile phase Sample solution: Sample stock solution and Mobile phase (1:5) Stock solution A: 1 mg/mL of USP Amitriptyline Related Compound A RS in methanol Stock solution B: Dissolve USP Amitriptyline Hydrochloride RS, USP Amitriptyline Related Compound B RS, USP Cyclobenzaprine Hydrochloride RS, and USP Nortriptyline Hydrochloride RS in Mobile phase to obtain a solution having a known concentration of 0.4 mg/mL of amitriptyline hydrochloride and 0.6 mg/mL each of amitriptyline related compound B, cyclobenzaprine hydrochloride, and nortriptyline hydrochloride. System suitability solution: Dilute suitable volumes of Stock solution A, Stock solution B, and the Standard solution with Mobile phase to obtain a solution having 1 g/mL of amitriptyline hydrochloride, 0.5 g/mL of amitriptyline related compound A, and 1.5 g/mL each of amitriptyline related compound B, cyclobenzaprine hydrochloride, and nortriptyline hydrochloride. Chromatographic system (See Chromatography 621, System Suitability.)
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CU
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Acceptance criteria: The UV absorption spectrum of this solution exhibits a maximum at the same wavelength as that of a similar solution of USP Amitriptyline Hydrochloride RS, concomitantly measured. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Analysis: Pipet a volume of Injection, equivalent to 50 mg of amitriptyline hydrochloride, into a separator containing 25 mL of water. Proceed as directed in the chapter, beginning with In a second separator, and using water in place of 0.01 N hydrochloric acid in the Reference Standard solution. Acceptance criteria: The Sample solution so obtained meets the requirements of the test. ASSAY PROCEDURE Buffer: 11.04 g of monobasic sodium phosphate in 900 mL of water, adjust with phosphoric acid to a pH of 2.5 0.5, and dilute with water to make 1000 mL. Mobile phase: Acetonitrile and Buffer (21:29) Standard solution: 0.2 mg/mL of USP Amitriptyline Hydrochloride RS Sample solution: Nominally equivalent to 0.2 mg/mL of amitriptyline hydrochloride, from Injection diluted with water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 800 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Sample: Sample solution and Standard solution Calculate the percentage of C20H23N HCl in each mL of Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration, in mg/mL, of USP Amitriptyline Hydrochloride RS in the Standard solution CU = nominal concentration of amitriptyline hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PYROGEN 151: Amitriptyline Hydrochloride Injection, diluted with Sodium Chloride Injection containing 0.9% of NaCl to a concentration of 2.5 mg of amitriptyline hydrochloride/mL, meets the requirements, the test dose being 1 mL/kg. PH 791: 4.06.0 OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Amitriptyline Hydrochloride RS rU rS CS
= concentration of amitriptyline hydrochloride in the Sample solution (mg/mL) [NOTEDiscard any peak with a relative retention time of less than 0.22. Use the response of the amitriptyline hydrochloride peak of the Standard solution and the concentration of amitriptyline hydrochloride in the Standard solution to calculate the percentage of unknown individual impurities.] Acceptance criteria: See Impurity Table 1.
Impurity Table 1 Relative Retention Time 0.35 Relative Response Factor Acceptance Criteria, NMT (%) 0.05
Name Amitriptyline related compound A Amitriptyline related compound B Nortriptyline hydrochloride Cyclobenzaprine hydrochloride Amitriptyline hydrochloride Any unspecified impurity Total impurities
0.52
0.15
0.60 0.76 1.0
0.15 0.15 0.10 each 1.0
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 195199 PH 791: 5.06.0, in a solution (1 in 100) LOSS ON DRYING 731: Dry a sample at a pressure not exceeding 5 mm of mercury at 60 to constant weight: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amitriptyline Hydrochloride RS USP Amitriptyline Related Compound A RS USP Amitriptyline Related Compound B RS USP Cyclobenzaprine Hydrochloride RS USP Nortriptyline Hydrochloride RS
Amitriptyline Hydrochloride Injection

(Comment on this Monograph)id=m3510=Amitriptyline Hydrochloride Injection=A-Monos.pdf) DEFINITION Amitriptyline Hydrochloride Injection is a sterile solution of Amitriptyline Hydrochloride in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of amitriptyline hydrochloride (C20H23N HCl). IDENTIFICATION A. UV ABSORPTION Sample solution: Pipet 1 mL of Injection into a 125-mL separator containing 10 mL of water and 1 mL of 1 N sodium hydroxide, mix, extract with two 10-mL portions of methylene chloride, and evaporate the extracts on a steam bath just to dryness. Dissolve the residue in methanol, add 1 mL of 1.2 N hydrochloric acid, then add methanol to make 100 mL. Dilute 10 mL of this solution with methanol to 100 mL.
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Official Monographs / Amlodipine 209

Amitriptyline Hydrochloride Tablets

(Comment on this Monograph)id=m3550=Amitriptyline Hydrochloride Tablets=A-Monos.pdf) DEFINITION Amitriptyline Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of amitriptyline hydrochloride (C20H23N HCl). IDENTIFICATION A. PROCEDURE Analysis: Transfer a quantity of finely powdered Tablets, equivalent to 10 mg of amitriptyline hydrochloride, to a 100-mL volumetric flask, add 50 mL of methanol, shake well, then add methanol to volume. Filter a portion of this solution, and dilute 10 mL of the filtrate with methanol to 100 mL. Acceptance criteria: The UV absorption spectrum of this solution exhibits a maximum at the same wavelength as that of a similar solution of USP Amitriptyline Hydrochloride RS, concomitantly measured. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: 11.04 g of monobasic sodium phosphate in 900 mL of water, adjust with phosphoric acid to a pH of 2.5 0.5, dilute to make 1000 mL Mobile phase: Acetonitrile and Buffer (21:29) Standard solution: 0.2 mg/mL of USP Amitriptyline Hydrochloride RS in Mobile phase Sample solution: Transfer 20 Tablets to a 500-mL volumetric flask, add 250 mL of Mobile phase, and shake the mixture for 1 h or until the tablets have disintegrated. Add Mobile phase to volume and filter. Dilute a measured volume of the clear filtrate with Mobile phase to obtain a solution containing 0.2 mg/mL of amitriptyline hydrochloride. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 800 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C20H23N HCl in each Tablet taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amitriptyline Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of amitriptyline hydrochloride in the Sample solution (mg/mL)
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 45 min Spectrometric conditions Analytical wavelength: UV 239 nm Standard solution: USP Amitriptyline Hydrochloride RS in Medium Sample solution: Sample per Dissolution 711 Analysis: Determine the amount of C20H23N HCl dissolved from UV absorbances of the Sample solution, suitably diluted with Medium, if necessary, in comparison with a Standard solution having a known concentration. Tolerances: NLT 75% (Q) of the labeled amount of C20H23N HCl UNIFORMITY OF DOSAGE UNITS: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amitriptyline Hydrochloride RS
Amlodipine Besylate
(Comment on this Monograph)id=m3570=Amlodipine Besylate=A-Monos.pdf)
567.05 C20H25ClN2O5 C6H6O3S 3,5-Pyridinedicarboxylic acid, 2-[(2-aminoethoxy)methyl]-4-(2chlorophenyl)-1,4-dihydro-6-methyl-, 3-ethyl 5-methyl ester, ()-, monobenzenesulfonate; 3-Ethyl 5-methyl ()-2-[(2-aminoethoxy)methyl]-4-(ochlorophenyl)-1,4-dihydro-6-methyl-3,5-pyridinedicarboxylate, monobenzenesulfonate [111470-99-6]. DEFINITION Amlodipine Besylate contains NLT 97.0% and NMT 102.0% of C20H25ClN2O5 C6H6O3S, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: Dissolve 7.0 mL of triethylamine in 800 mL of water. Adjust with phosphoric acid to a pH of 3.0 0.1, and dilute with water to 1 L. Mobile phase: Methanol, acetonitrile, and Buffer (7:3:10) Standard solution: 0.05 mg/mL of USP Amlodipine Besylate RS in Mobile phase
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System suitability solution: Dissolve 5 mg of Amlodipine Besylate in 5 mL of hydrogen peroxide, and heat at 70 for 45 min. Standard solution: 0.003 mg/mL of USP Amlodipine Besylate RS in Mobile phase Sample solution: 1 mg/mL of Amlodipine Besylate in Mobile phase Chromatographic system: Proceed as directed in the Assay. (See Chromatography 621, System Suitability.) System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for benzene sulfonate, amlodipine impurity A, and amlodipine are 0.2, 0.5, and 1.0, repectively. Amlodipine impurity A is 3-ethyl 5-methyl 2-[(2-aminoethoxy)methyl]-4-(2chlorophenyl)-6-methylpyridine-3,5-dicarboxylate.] Suitability requirements Resolution: NLT 4.5 between amlodipine impurity A and amlodipine, System suitability solution Relative standard deviation: NMT 10%, Standard solution Analysis Samples: Standard solution and Sample solution Record the chromatograms for a period of time that is 3 times the retention time of amlodipine. Calculate the percentage of each impurity in the portion of Amlodipine Besylate taken: Result = (rU/rS) (CS/CU) (1/F) 100 = peak response for each impurity from the Sample solution rS = peak response for amlodipine besylate from the Standard solution = concentration of USP Amiodipine Besylate RS in CS the Standard solution (mg/mL) = concentration of amiodipine besylate in the CU Sample solution (mg/mL) F = relative response factor, 0.5 for amlodipine impurity A and to 1.0 for other impurities Acceptance criteria Amlodipine impurity A: NMT 0.3% Total other impurities: NMT 0.3% [NOTEDisregard any peak less than 0.03%, and disregard any peak due to benzene sulfonate.] SPECIFIC TESTS OPTICAL ROTATION 781: 0.10 to +0.10, measured at 20 Sample solution: 10 mg/mL, in methanol WATER DETERMINATION, Method I 921: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. Store at room temperature. USP REFERENCE STANDARDS 11 USP Amlodipine Besylate RS rU
Sample solution: 0.05 mg/mL of Amlodipine Besylate in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 237 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1.0 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C20H25ClN2O5 C6H6O3S in the portion of Amlodipine Besylate taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amiodipine Besylate RS in the Standard solution (mg/mL) = concentration of amiodipine besylate in the CU Sample solution (mg/mL) Acceptance criteria: 97.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1 Standard stock solution: 7 mg/mL of USP Amlodipine Besylate RS in methanol Standard solution 1: 0.21 mg/mL from Standard stock solution and methanol Standard solution 2: 0.07 mg/mL from Standard stock solution and methanol Sample solution: 70 mg/mL of Amlodipine Besylate in methanol System suitability solution: 70 mg of USP Amlodipine Besylate RS in methanol Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Use the upper layer of a mixture of methyl isobutyl ketone, water, and glacial acetic acid (2:1:1). Analysis Samples: Standard solution 1 and 2, Sample solution, and System suitability solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Dry the plate for 15 min at 80. Examine the plate under UV light at 254 nm and at 365 nm. The chromatogram from the System suitability solution shows two clearly separated minor spots with RF values of 0.18 and 0.22. Compare the intensities of any secondary spots observed in the chromatogram of the Sample solution with those of the principal spots in the chromatograms of the Standard solutions. Acceptance criteria: Any spot obtained from the Sample solution, except for the principal spot, is not greater in size than the spot obtained from Standard solution 1 (0.3%), and at most two spots are more intense than the spot obtained from Standard solution 2 (0.1%). PROCEDURE 2 Buffer and Mobile phase: Prepare as directed in the Assay.
Aromatic Ammonia Spirit

(Comment on this Monograph)id=m3630=Aromatic Ammonia Spirit=A-Monos.pdf) DEFINITION Aromatic Ammonia Spirit is a hydroalcoholic solution that contains, in each 100 mL, NLT 1.7 g and NMT 2.1 g of total NH3 and ammonium carbonate corresponding to NLT 3.5 g and NMT 4.5 g of (NH4)2CO3.
USP 32
ASSAY TOTAL NH3 Sample: 10.0 mL Analysis: Transfer to a 250-mL conical flask containing 50 mL of water. Add 30.0 mL of 0.5 N sulfuric acid VS, and boil until the solution becomes clear. Cool, and add methyl red TS. Titrate the excess acid with 0.5 N sodium hydroxide VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.5 N sulfuric acid is equivalent to 8.515 mg of NH3. AMMONIUM CARBONATE Sample: 10.0 mL Analysis: Transfer to a flask of 300-mL capacity. Add 30 mL of 0.5 N sodium hydroxide, and boil the mixture, replacing the water lost by evaporation, until the vapors no longer turn moistened red litmus paper blue. Cool, dilute with 100 mL of cold, carbon dioxide-free water, add 6 drops of phenolphthalein TS, then add just enough 0.5 N sulfuric acid VS to discharge the color of the phenolphthalein. Add methyl orange TS. Titrate with 0.5 N sulfuric acid VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.5 N sulfuric acid consumed in the titration with methyl orange TS is equivalent to 48.04 mg of (NH4)2CO3. Acceptance criteria: In each 100 mL, NLT 1.7 g and NMT 2.1 g of total NH3 and ammonium carbonate corresponding to NLT 3.5 g and NMT 4.5 g of (NH4)2CO3 OTHER COMPONENTS ALCOHOL DETERMINATION, Method I 611: 62.0%68.0%
Official Monographs / Ammonium 211

HEAVY METALS, Method I 231: NMT 10 ppm LIMIT OF THIOCYANATE Sample solution: 100 mg/mL Analysis: Acidify 10 mL of the Sample solution with hydrochloric acid, and add a few drops of ferric chloride TS. Acceptance criteria: No orange-red color is produced. SPECIFIC TESTS PH 791 Sample solution: 50 mg/mL Acceptance criteria: 4.66.0 LOSS ON DRYING 731: Dry over silica gel for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Ammonium Chloride Injection

(Comment on this Monograph)id=m3720=Ammonium Chloride Injection=A-Monos.pdf) DEFINITION Ammonium Chloride Injection is a sterile solution of Ammonium Chloride in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of NH4Cl. Hydrochloric acid may be added to adjust the pH. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Ammonium 191: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Sample: A volume of Injection equivalent to 200 mg of ammonium chloride Analysis: Transfer Sample to a 500-mL Kjeldahl flask. Dilute with water to 200 mL, and add 50 mL of sodium hydroxide solution (2 in 5). Immediately connect the flask by means of a distillation trap to a well-cooled condenser, the delivery tube of which dips into 40 mL of boric acid solution (1 in 25) contained in a suitable receiver. Heat to boiling, and distill 200 mL. Cool the liquid in the receiver, if necessary, then add methyl red TS. Titrate with 0.1 N sulfuric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N sulfuric acid is equivalent to 5.349 mg of NH4Cl. Acceptance criteria: 95.0%105.0% OTHER COMPONENTS CHLORIDE CONTENT Sample solution: A measured volume of Injection, evaporated, if necessary, equivalent to 2 g of ammonium chloride Analysis: Transfer Sample solution to a 200-mL volumetric flask. Dilute with water to volume. Transfer 10.0 mL of this solution to a conical flask, add 10 mL of glacial acetic acid, 75 mL of methanol, and 0.5 mL of eosin Y TS. Titrate, with shaking, with 0.1 N silver nitrate VS to a pink endpoint. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Cl. Acceptance criteria: 63.0%70.3% SPECIFIC TESTS PH 791 Sample solution: A concentration of NMT 100 mg/mL of ammonium chloride
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at a temperature not exceeding 30.
Ammonium Chloride
(Comment on this Monograph)id=m3690=Ammonium Chloride=A-Monos.pdf) NH4Cl Ammonium chloride [12125-02-9]. 53.49
DEFINITION Ammonium Chloride contains NLT 99.5% and NMT 100.5% of NH4Cl, calculated on the dried basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Ammonium and Chloride 191: Meets the requirements Sample solution: 100 mg/mL ASSAY PROCEDURE Sample: 100 mg of Ammonium Chloride Analysis: Add 10 mL of water to the Sample, and swirl to dissolve. Add 10 mL of glacial acetic acid, 75 mL of methanol, and 0.5 mL of eosin Y TS. Titrate, with shaking, with 0.1 N silver nitrate VS to a pink endpoint. Each mL of 0.1 N silver nitrate is equivalent to 5.349 mg of NH4Cl. Acceptance criteria: 99.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Sample: 2 g Analysis: Add 1 mL of sulfuric acid to the Sample, and heat the mixture gently until volatilization is complete. Ignite the residue. Acceptance criteria: The residue is white, and when ignited, NMT 0.1% of nonvolatile substance remains.
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No reddish-orange color is produced.
Acceptance criteria: 4.06.0 OTHER REQUIREMENTS: It meets the requirements under Injections 1. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.72 USP Endotoxin Units/mEq of chloride. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I or Type II glass. LABELING: The label states the content of ammonium chloride in terms of weight and milliequivalents in a given volume. The label states also the total osmolar concentration in mOsmol/L or mOsmol/mL. The label states that the Injection is not for direct injection but is to be diluted with Sodium Chloride Injection to the appropriate strength before use. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Ferric Ammonium Citrate

(Comment on this Monograph)id=m3761=Ferric Ammonium Citrate=A-Monos.pdf) DEFINITION Ferric Ammonium Citrate contains NLT 16.5% and NMT 18.5% of iron (Fe). IDENTIFICATION A. Ignite 0.5 g: it chars, and leaves a residue of iron oxide. B. To 10 mL of a solution of Ferric Ammonium Citrate (1 in 100) add 6 N ammonium hydroxide dropwise: the solution darkens, but no precipitate forms. C. To 5 mL of a solution of Ferric Ammonium Citrate (1 in 100) add 0.3 mL of potassium permanganate TS and 4 mL of mercuric sulfate TS, and heat the mixture to boiling: a white precipitate forms. ASSAY PROCEDURE Sample solution: Transfer 1 g of Ferric Ammonium Citrate to a 250-mL conical flask, and dissolve in 25 mL of water and 5 mL of hydrochloric acid. Analysis: Add 4 g of potassium iodide, insert the stopper, and allow to stand protected from light for 15 min. Add 100 mL of water. Titrate the liberated iodine with 0.1 N sodium thiosulfate VS, using starch TS as the indicator. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N sodium thiosulfate is equivalent to 5.585 mg of iron (Fe). Acceptance criteria: 16.5%18.5% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Sulfate 221 Sample solution: Dissolve 100 mg in 1 mL of 2.7 N hydrochloric acid, and dilute with water to 30 mL. Add 3 mL of barium chloride TS, and dilute with water to 50 mL. Acceptance criteria: Any turbidity formed after 10 min is NMT that produced in a similarly treated control solution containing 0.31 mL of 0.020 N sulfuric acid (0.3%). LIMIT OF LEAD Standard stock solution: Dissolve 159.8 mg of lead nitrate in 100 mL of water containing 1 mL of nitric acid. Dilute with water to 1000 mL. Standard solution: [NOTEPrepare this solution on the day of use.] Transfer 10 mL of Standard stock solution to a 500-mL volumetric flask, and dilute with water to volume. Each mL contains the equivalent of 2 g of lead (Pb). Sample solution: Transfer 15 g of Ferric Ammonium Citrate to a 100-mL volumetric flask (previously rinsed with nitric acid and water), dissolve in a mixture of 50 mL of water and 1 mL of nitric acid, and dilute with water to volume. Analysis Samples: Standard solution and Sample solution Blank: Water Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer equipped with a deuterium arc background corrector, a digital readout device, and a burner head capable of handling 15% solids content
Ammonium Chloride Delayed-Release Tablets

(Comment on this Monograph)id=m3760=Ammonium Chloride Delayed-Release Tablets=A-Monos.pdf) DEFINITION Ammonium Chloride Delayed-Release Tablets contain NLT 94.0% and NMT 106.0% of the labeled amount of NH4Cl. Ammonium Chloride Delayed-Release Tablets are entericcoated. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Ammonium and Chloride 191: Meets the requirements Sample solution: A filtered solution of finely powdered Tablets, equivalent to 100 mg/mL ammonium chloride ASSAY PROCEDURE Sample: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 200 mg of ammonium chloride, to a 500-mL Kjeldahl flask. Analysis: Add 200 mL of water and 50 mL of sodium hydroxide solution (2 in 5) to the Sample. Immediately connect the flask by means of a distillation trap to a wellcooled condenser, the delivery tube of which dips into 40 mL of boric acid solution (1 in 25) contained in a suitable receiver. Heat to boiling, and distill 200 mL. Cool the liquid in the receiver, if necessary, then add methyl red TS. Titrate with 0.1 N sulfuric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N sulfuric acid is equivalent to 5.349 mg of NH4Cl. Acceptance criteria: 94.0%106.0% PERFORMANCE TESTS DISINTEGRATION 701: Enteric-Coated Tablets 2 h, determined as directed for
IMPURITIES Inorganic Impurities LIMIT OF THIOCYANATE Sample solution: Powder and dissolve in water a sufficient number of Tablets to make 25 mL of a solution containing 100 mg/mL of ammonium chloride solution, and filter. Analysis: Acidify 10 mL of the Sample solution with hydrochloric acid, and add a few drops of ferric chloride TS.
USP 32
Analysis Samples: Standard solution and Sample solution Perform a blank determination with water, following the manufacturers operating instructions. Separately aspirate portions of the Standard solution and the Sample solution, and record the absorbances. Calculate the lead content, in g per g, in the portion of Ferric Ammonium Citrate taken: (AU/AS) CS/CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of lead in the Standard solution (g/mL) CU = concentration of Ferric Ammonium Citrate (g/ml) Acceptance criteria: NMT 10 g/g MERCURY Mercury Stock Solution and Standard Mercury solution: Proceed as directed for Mercury 261, Method I. Mercury Detection Instrument, Aeration Apparatus, and Stannous Chloride Solution: Proceed as directed for Mercury 261, Method IIa and Method IIb. Standard solutions: Transfer 0.25, 0.50, 1.0, and 3.5 mL of Mercury Stock Solution to four separate glass-stoppered bottles, such as biological oxygen-demand bottles, of about 300-mL capacity. Dilute the contents of each bottle with water to 100 mL. These solutions contain the equivalent of 2.5, 5.0, 10.0, and 35.0 g/mL of mercury, respectively. Sample solution: Transfer 1.000 g of Ferric Ammonium Citrate to a 200-mL centrifuge bottle with a polytef-lined screw cap, and add 5 mL of nitric acid and 5 mL of hydrochloric acid. Close the bottle tightly, digest on a steam bath for 1 h, and cool. Quantitatively transfer the solution to a suitable glass-stoppered bottle, dilute with water to 100 mL, and bubble air through the solution for 2 min. Prepare a reagent blank in the same manner. Analysis Samples: Standard solution and Sample solution Add 5 mL of Stannous Chloride Solution (1 in 10) to each solution, and immediately insert the bubbler of the Aeration Apparatus. Obtain the absorbances as directed by the instrument manufacturers operating instructions. Perform a blank determination, and make any necessary correction. Plot the absorbances of the Standard solutions versus concentrations, in g/mL, of mercury, and draw the straight line best fitting the plotted points. From the graph so obtained, determine the concentration, in g/g, of mercury in the Sample solution. Acceptance criteria: NMT 10 g/g AU AS CS SPECIFIC TESTS FERRIC CITRATE: To a solution of Ferric Ammonium Citrate (1 in 100) add potassium ferrocyanide TS: no blue precipitate is formed. OXALATE: Transfer 1 g to a 125-mL separator, dissolve in 10 mL of water, add 2 mL of hydrochloric acid, and extract successively with one 50-mL portion and one 20-mL portion of ether. Transfer the combined ether extracts to a 150-mL beaker, add 10 mL of water, and remove the ether by evaporation on a steam bath. Add 1 drop of glacial acetic acid and 1 mL of calcium acetate solution (1 in 20): no turbidity is produced within 5 min. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, in a cool place.
Official Monographs / Ammonium 213
Ferric Ammonium Citrate for Oral Solution

(Comment on this Monograph)id=m3762=Ferric Ammonium Citrate for Oral Solution=A-Monos.pdf) DEFINITION Ferric Ammonium Citrate for Oral Solution contains Ferric Ammonium Citrate and an effervescent mixture of a suitable organic acid and an alkali metal bicarbonate. It contains NLT 90.0% and NMT 110.0% of the labeled amount of Fe. It may contain one or more suitable flavors, colors, or stabilizing agents. IDENTIFICATION A 6-g portion dissolves in 600 mL of water with effervescence. The collected gas meets the requirements of the test for Identification TestsGeneral, Bicarbonate 191, and the resulting solution meets the requirements of the tests for Identification TestsGeneral, Iron 191, and for Identification TestsGeneral, Citrate 191. ASSAY PROCEDURE Sample solution: Transfer 6 g of Ferric Ammonium Citrate for Oral Solution to a 250-mL conical flask, and dissolve in 100 mL of water. Allow the gas to escape, add 5 mL of hydrochloric acid and 4 g of potassium iodide, insert the stopper, and allow to stand protected from light for 15 min. Add 25 mL of water. Analysis: Titrate the liberated iodine with 0.1 N sodium thiosulfate VS, using starch TS as the indicator. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N sodium thiosulfate is equivalent to 5.585 mg of Fe. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: It meets the requirements for powder packaged in single-unit containers. SPECIFIC TESTS DELIVERABLE VOLUME 698: It meets the requirements for powder packaged in multiple-unit containers. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store in a cool place.
Ammonium Molybdate
(Comment on this Monograph)id=m3795=Ammonium Molybdate=A-Monos.pdf) 1235.86 (NH4)6Mo7O24 4H2O Molybdate (Mo7O246), hexaammonium, tetrahydrate; Hexaammonium molybdate tetrahydrate [12054-85-2] DEFINITION Ammonium Molybdate contains NLT 99.3% and NMT 101.8% of (NH4)6Mo7O24 4H2O. IDENTIFICATION PROCEDURE Sample: 0.6 g Analysis: Dissolve the Sample in 1.4 mL of water and 1.45 mL of ammonium hydroxide. Cool this mixture, and add slowly, with mixing, 7.2 mL of a well-cooled mixture of 3.2 mL of nitric acid and 4 mL of water. Allow to stand for 24
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INSOLUBLE SUBSTANCES Sample: 20 g Analysis: Dissolve the Sample in 200 mL of water in a beaker, heat to boiling, cover, and heat on a steam bath for 1 h. Filter the hot solution through a tared filtering crucible, wash the insoluble residue with hot water, and dry at 105 for 2 h. Acceptance criteria: The weight of the residue so obtained is NMT 1 mg (0.005%). LIMIT OF NITRATE Sample: 1 g Analysis: Dissolve the Sample in 10 mL of water containing 5 mg of sodium chloride, and add 0.10 mL of a 1 in 1000 solution of indigo carmine in 3.6 N sulfuric acid. Acceptance criteria: The blue color is not completely discharged in 5 min. ARSENATE, PHOSPHATE, AND SILICATE Sample: 2.5 g Analysis: Dissolve the Sample in 70 mL of water in a container other than one of glass. Adjust with 1.2 N hydrochloric acid to a pH of between 3 and 4, transfer to a glass container, add 2 mL of bromine TS, and adjust with 1.2 N hydrochloric acid to a pH of 1.8 0.1. Heat almost to boiling, and cool to room temperature. Dilute with water to 90 mL, add 10 mL of hydrochloric acid, and transfer to a separator. Add 1 mL of butyl alcohol and 30 mL of 4-methyl-2-pentanone, shake vigorously, and allow the phases to separate. Discard the aqueous phase, and wash the ketone phase with three successive 10-mL portions of 1.2 N hydrochloric acid, discarding the washings. To the washed ketone phase, add 10 mL of 1.2 N hydrochloric acid to which has just been added 0.2 mL of a freshly prepared 1 in 50 solution of stannous chloride in hydrochloric acid. Similarly treat a control solution prepared by dissolving 0.5 g of specimen in 70 mL of water and adding 2 mL of sodium silicate solution (1 in 20,000). Acceptance criteria: Any blue color in the test solution does not exceed that in the control solution. MAGNESIUM AND ALKALI SALTS Sample: 5.0 g Analysis: Dissolve the Sample in 50 mL of water, and filter. To the filtrate, add 0.5 g of sodium carbonate and 25 mL of 2.5 N sodium hydroxide. Boil the solution gently for 5 min, cool, and filter through an ignited and tared filter. Wash the filter with 1 N ammonium hydroxide. Ignite the filter at 800 25 for 30 min. Acceptance criteria: The weight of the residue so obtained does not exceed 1 mg (NMT 0.200 ppm). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
to 48 h, and filter through a sintered-glass filter. To 5 mL of the filtrate, add 2 mL of dibasic sodium phosphate TS. Acceptance criteria: A yellow precipitate is formed, and it is soluble in an excess of 6 N ammonium hydroxide. ASSAY PROCEDURE Sample solution: 1 g Analysis: Dissolve the Sample solution in a mixture of 10 mL of water and 1 mL of ammonium hydroxide in a 250-mL volumetric flask, and dilute with water to volume. Filter the solution, and transfer 50.0 mL of the filtrate to a 600-mL beaker. Add 250 mL of water, 20 g of ammonium chloride, 15 mL of hydrochloric acid, and 0.15 mL of methyl orange TS, heat nearly to boiling, and add 18 mL of lead acetate TS. To the hot solution add, slowly and with constant stirring, a saturated solution of ammonium acetate until the color turns yellow, and then add 15 mL of lead acetate TS. Cover the beaker, and heat just below the boiling temperature until the precipitate has settled. Filter through a tared, porous porcelain crucible, wash with seven or eight successive portions of a mixture of water, saturated ammonium acetate solution, and nitric acid (890:100:10), and finally wash with three successive portions of hot water. Ignite to constant weight at 560 to 625. Each mg of the lead molybdate so obtained is equivalent to 0.4809 mg of (NH4)6Mo7O24 4H2O. Acceptance criteria: 99.3%101.8% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 0.5-g portion shows no more chloride than 0.30 mL of 0.001 N hydrochloric acid (NMT 20 ppm). CHLORIDE AND SULFATE, Sulfate 221: A 0.25-g portion shows no more sulfate than corresponds to 1.0 mL of 0.001 N sulfuric acid (NMT 200 ppm). HEAVY METALS 231 Stock solution: Dissolve 2.0 g Ammonium Molybdate in 20 mL of water, add 10 mL of 2.5 N sodium hydroxide and 2 mL of ammonium hydroxide, and dilute with water to 40 mL. Control solution: To 10 mL of Stock solution add 1.0 mL of Standard Lead Solution, prepared as directed under Heavy Metals 231, and dilute with water to 40 mL. Sample solution: Dilute the remaining 30-mL portion of the Stock solution with water to 40 mL. Analysis: To both solutions, add 1.2 mL of thioacetamideglycerin base TS and 2 mL of pH 3.5 Acetate Buffer, and allow to stand for 5 min: any color in the Sample solution does not exceed that in the Control solution. Acceptance criteria: NMT 10 ppm PHOSPHATE Sample: 20 g Analysis: Dissolve the Sample in 100 mL of 3 N ammonium hydroxide, add 3.5 mL of ferric nitrate solution (1 in 10), and allow to stand for 15 min. Warm gently to coagulate the precipitate, and filter. Wash the filter several times with 1.5 N ammonium hydroxide, then wash the filter with 60 mL of warm 4 N nitric acid to dissolve the residue on the filter, collecting the filtrate in a glassstoppered, 250-mL conical flask. Add 13 mL of ammonium hydroxide, warm to 40, add 50 mL of ammonium molybdate TS, shake for 5 min, and allow to stand at 40 for 2 h. Similarly treat 100 mL of a Standard solution prepared by dissolving 143.3 mg of dried monobasic potassium phosphate in water to make 1000 mL, and then diluting 1.0 mL of this solution with 3 N ammonium hydroxide to 100 mL. Acceptance criteria: Any yellow precipitate formed from the Sample solution does not exceed that obtained from the Standard solution (5 ppm).
Ammonium Molybdate Injection

(Comment on this Monograph)id=m3798=Ammonium Molybdate Injection=A-Monos.pdf) DEFINITION Ammonium Molybdate Injection is a sterile solution of Ammonium Molybdate in Water for Injection. It contains NLT 85.0% and NMT 115.0% of the labeled amount of molybdenum (Mo). IDENTIFICATION A. The Sample solution, prepared as directed in the Assay, exhibits an absorption maximum at about 313 nm when tested as directed for Analysis in the Assay.
USP 32
B. PROCEDURE Sample: 5 mL of Injection Analysis: Add 0.3 mL of alkaline mercuric-potassium iodide TS to the Sample. Acceptance criteria: A reddish-brown color develops. C. PROCEDURE Sample: 50 mL of Injection Analysis: Evaporate the Sample on a steam bath to a volume of 0.3 mL, and add 0.3 mL of ammonium hydroxide. Cool, and add slowly, with mixing, a well-cooled mixture of 1 mL of nitric acid and 1.2 mL of water. Allow to stand for 2448 h, and pass through a sintered-glass filter. To the filtrate add 0.5 mL of dibasic sodium phosphate TS. Acceptance criteria: A yellow precipitate is formed, and it dissolves in an excess of 6 N ammonium hydroxide. ASSAY PROCEDURE Diluent: Ammonium hydroxide and water (1:24) [NOTEStore in a plastic bottle.] Solution A: 10 mg/mL of sodium sulfate. Molybdenum stock solution: Transfer 1.84 g of previously assayed Ammonium Molybdate to a 1000-mL volumetric flask, and dilute with Diluent to volume. This solution contains the equivalent of 1000 g/mL of molybdenum. Standard solutions: Transfer 0, 1, 2, 3, and 4 mL, respectively, of Molybdenum stock solution to separate 100mL volumetric flasks, and to the respective flasks add 5, 4, 3, 2, and 1 mL of Diluent. To each flask add 10 mL of Solution A, and dilute with water to volume. These Standard solutions contain, respectively, 0, 10, 20, 30, and 40 g of molybdenum/mL. Sample solution: A volume of Injection, equivalent to 500 g of molybdenum, to a 25-mL volumetric flask. Add 1.25 mL of Diluent and 2.5 mL of Solution A, and dilute with water to volume. Spectrometric conditions Mode: Atomic absorption spectrophotometer (See Spectrophotometry and Light-Scattering 851.) Analytical wavelength: Molybdenum emission line of 313.3 nm Lamp: Molybdenum hollow-cathode lamp Flame: Nitrous oxideacetylene reducing flame Blank: Water Analysis Samples: Standard solutions and the Sample solution Plot the absorbances of the Standard solutions versus concentration, in g/mL, of molybdenum, and draw the straight line best fitting the five plotted points. From the graph so obtained, determine the concentration, in g/mL, of molybdenum in the Sample solution. Calculate the percentage of Mo in each mL of the Injection taken: Result = 25 (C/V) 100 = concentration of molybdenum in the Sample solution (g/mL) V = volume of Injection taken (mL) Acceptance criteria: 85.0%115.0% SPECIFIC TESTS PYROGEN TEST 151: Meets the requirements, the test dose being 10 mL of Injection/kg PH 791: 3.06.0 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: Meets the requirements under Injections 1 C
Official Monographs / Amobarbital 215

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I or Type II glass. LABELING: Label the Injection to indicate that it is to be diluted to the appropriate strength with Sterile Water for Injection or other suitable fluid before administration.
Amobarbital Sodium
(Comment on this Monograph)id=m3950=Amobarbital Sodium=A-Monos.pdf) C11H17N2NaO3 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-5-(3-methylbutyl)-, monosodium salt; Sodium 5-ethyl-5-isopentylbarbiturate [64-43-7]. 248.25
DEFINITION Amobarbital Sodium contains NLT 98.5% and NMT 100.5% of C11H17N2NaO3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Residue obtained in the Assay B. IDENTIFICATION TESTSGENERAL, Sodium 191 Sample: 200 mg Analysis: Ignite Sample Acceptance criteria: The residue effervesces with acid and responds to the test for sodium. ASSAY PROCEDURE Sample solution: 500 mg of Amobarbital Sodium, in 15 mL of water in a separator Analysis: To the Sample solution add 2 mL of hydrochloric acid, shake, and completely extract the liberated amobarbital with 25-mL portions of chloroform. Test for completeness of extraction by extracting with an additional 10-mL portion of chloroform and evaporating the solvent: NMT 0.5 mg of residue remains. Filter the combined extract through a glass filter funnel into a tared beaker, and wash the separator and the filter with several small portions of chloroform. Evaporate the combined filtrate and washings on a steam bath with the aid of a current of air, dry the residue at 105 for 30 min, cool, and weigh. The weight of the residue, multiplied by 1.097, represents the weight of C11H17N2NaO3. Acceptance criteria: 98.5%100.5% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231:
NMT 30 ppm
SPECIFIC TESTS COMPLETENESS OF SOLUTION 641: 1.0 g in 10 mL of carbon dioxide-free water: after 1 min, the solution is clear and free from undissolved solid. PH 791: NMT 11.0, in the solution prepared for the test for Completeness of solution LOSS ON DRYING 731: Dry 1 g, at 105 for 4 h: it loses NMT 2.0% of its weight. OTHER REQUIREMENTS: Where the label states that Amobarbital Sodium is sterile, it meets the requirements for Sterility Tests 71 and it contains NMT 0.4 USP Endotoxin Unit/mg of amobarbital sodium (Bacterial Endotoxins Test 85.) Where the label states that Amobarbital Sodium must be subjected to further processing during the preparation of
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Amobarbital RS USP Endotoxin RS
injectable dosage forms, it meets the above requirements for Bacterial Endotoxins. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Amobarbital RS USP Endotoxin RS
Amodiaquine
(Comment on this Monograph)id=m4020=Amodiaquine=AMonos.pdf)
Amobarbital Sodium for Injection

(Comment on this Monograph)id=m3960=Amobarbital Sodium for Injection=A-Monos.pdf) DEFINITION Amobarbital Sodium for Injection is Amobarbital Sodium suitable for parenteral use. It contains NLT 98.5% and NMT 100.5% of the labeled amount of C11H17N2NaO3, calculated on the dried basis. ASSAY PROCEDURE Sample solution: A portion of Injection, equivalent to about 500 mg of amobarbital sodium, in 15 mL of water in a separator Analysis: To the solution, add 2 mL of hydrochloric acid, shake, and completely extract the liberated amobarbital with 25-mL portions of chloroform. Test for completeness of extraction by extracting with an additional 10-mL portion of chloroform and evaporating the solvent: NMT 0.5 mg of residue remains. Filter the combined extract through a glass filter funnel into a tared beaker, and wash the separator and the filter with several small portions of chloroform. Evaporate the combined filtrate and washings on a steam bath with the aid of a current of air, dry the residue at 105 for 30 min, cool, and weigh. The weight of the residue, multiplied by 1.097, represents the weight of C11H17N2NaO3. Acceptance criteria: 98.5%100.5% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: requirements IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: Meets the C20H22ClN3O 355.86 Phenol, 4-[(7-chloro-4-quinolinyl)amino]-2-[(diethylamino)methyl]-; 4-[(7-Chloro-4-quinolyl)amino]--(diethylamino)-o-cresol [86-42-0]. DEFINITION Amodiaquine contains NLT 97.0% and NMT 103.0% of C20H22ClN3O, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Standard solution: 20 mg of USP Amodiaquine Hydrochloride RS in 10 mL of water in a separator, add 1 mL of ammonium hydroxide, and extract by shaking with 25 mL of chloroform. Draw off and evaporate the chloroform extract, and dry the residue at 105 for 2 h. B. ULTRAVIOLET ABSORPTION 197U Sample solution: 10 g/mL in 0.1 N hydrochloric acid ASSAY PROCEDURE Standard solution: 15 g/mL of USP Amodiaquine Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: 15 g/mL of Amodiaquine in 0.1 N hydrochloric acid Spectrometric conditions Cell: 1 cm Analytical wavelength: 342 nm Blank: 0.1 N hydrochloric acid Analysis Samples: Sample solution and Standard solution Calculate the percentage of C20H22ClN3O in the portion of Amodiaquine taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 AU AS CS CU Mr1 Mr2 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Amodiaquine Hydrochloride RS in the Standard solution (g/mL) = concentration of amodiaquine hydrochloride in the Sample solution (g/mL) = molecular weight of amodiaquine, 355.87 = molecular weight of anhydrous aminodiaquine hydrochloride, 428.79
NMT 30 ppm
SPECIFIC TESTS INJECTIONS, Constituted Solutions 1: At the time of use, it meets the requirements. LOSS ON DRYING 731: Dry 1 g, at 105 for 4 h: it loses NMT 1.0% of its weight. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.4 USP Endotoxin Unit/mg of amobarbital sodium. STERILITY TESTS 71: Meets the requirements COMPLETENESS OF SOLUTION 641: 1.0 g in 10 mL of carbon dioxide-free water: after 1 min, the solution is clear and free from undissolved solid. PH 791: NMT 11.0, in the solution prepared for the Completeness of Solution 641 test. OTHER REQUIREMENTS: It meets the requirements of the Identification tests under Amobarbital Sodium. It meets the requirements under Injections 1, Labeling.
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B. ULTRAVIOLET ABSORPTION 197U Sample solution: 10 g/mL in dilute hydrochloric acid (1 in 100) C. IDENTIFICATION TESTSGENERAL, Chloride 191: meets the requirements. ASSAY PROCEDURE Sample solution: 15 g/mL of Amodiaquine Hydrochloride in dilute hydrochloric acid (1 in 100) Standard solution: 15 g/mL of USP Amodiaquine Hydrochloride RS in dilute hydrochloric acid (1 in 100) Spectrometric conditions Cell: 1 cm Analytical wavelength: 342 nm Blank: Dilute hydrochloric acid (1 in 100) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C20H22ClN3O 2HCl in the portion of Amodiaquine Hydrochloride taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Amodiaquine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of the Sample solution CU (g/mL) Acceptance criteria: 97.0%103.0% AU AS CS PERFORMANCE TESTS COMPLETENESS OF SOLUTION 641: 10 mL of water is clear. A solution of 200 mg in
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Standard solution A: To 20 mg of USP Amodiaquine Hydrochloride RS in a glass-stoppered test tube add 1.0 mL of chloroform (saturated with ammonium hydroxide), and shake vigorously for 2 min. Allow the solids to settle, and decant the liquid into a second test tube. Standard solution B: Standard solution A and chloroform (saturated with ammonium hydroxide) (1:199) Sample solution: 15 mg/mL of Amodiaquine in chloroform (saturated with ammonium hydroxide) Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform (saturated with ammonium hydroxide) and dehydrated alcohol (9:1) Analysis Samples: Standard solution A, Standard solution B, and and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the solvent to evaporate, and examine the plate under shortwavelength UV light. Acceptance criteria: The chromatograms show principal spots at about the same RF value, and no secondary spot, if present in the chromatogram from the Sample solution, is more intense than the principal spot obtained from Standard solution B. SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amodiaquine Hydrochloride RS
Amodiaquine Hydrochloride
(Comment on this Monograph)id=m4040=Amodiaquine Hydrochloride=A-Monos.pdf) 464.81 C20H22ClN3O 2HCl 2H2O Phenol, 4-[(7-chloro-4-quinolinyl)amino]-2-[(diethylamino)methyl]-, dihydrochloride, dihydrate; 4-[(7-Chloro-4-quinolyl)amino]--(diethylamino)-o-cresol dihydrochloride dihydrate [6398-98-7]. Anhydrous 428.79 [69-44-3]. DEFINITION Amodiaquine Hydrochloride contains NLT 97.0% and NMT 103.0% of C20H22ClN3O 2HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Dissolve 20 mg of Amodiaquine Hydrochloride in 10 mL of water in a separator, add 1 mL of ammonium hydroxide, and extract by shaking with 25 mL of chloroform. Draw off and evaporate the chloroform extract, and dry the residue at 105 for 2 h.
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Standard solution A: To 20 mg of USP Amodiaquine Hydrochloride RS in a glass-stoppered test tube, add 1.0 mL of chloroform (saturated with ammonium hydroxide), and shake vigorously for 2 min. Allow the solids to settle, and decant the liquid into a second test tube. Standard solution B: Dilute 1.0 mL of Standard solution A to 200 mL with chloroform (saturated with ammonium hydroxide). Sample solution: To 200 mg of Amodiaquine Hydrochloride in a glass-stoppered test tube, add 10 mL of chloroform (saturated with ammonium hydroxide). Shake vigorously for 2 min. Allow the solids to settle, and decant the liquid into a second test tube. Developing solvent system: Chloroform (saturated with ammonium hydroxide) and dehydrated alcohol (9:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Analysis Samples: Standard solution A, Standard solution B, and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the
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acid (1 in 100). Combine the acid extracts in a 200-mL volumetric flask, add dilute hydrochloric acid (1 in 100) to volume. Pipet 20 mL of this solution into a 100-mL volumetric flask, add dilute hydrochloric acid (1 in 100) to volume. Standard solution: 15 g/mL of undried USP Amodiaquine Hydrochloride RS in dilute hydrochloric acid (1 in 100) Spectrometric conditions Cell: 1 cm Analytical wavelength: 342 nm Blank: Dilute hydrochloric acid (1 in 100). Analysis Samples: Sample solution and Standard solution Calculate the percentage of C20H22ClN3O in the portion of Tablets taken: Result = (AU/AS) (CS/CU) F 100 = absorbance of the solution from the Tablets = absorbance of the solution from Standard solution = concentration calculated, on the anhydrous CS basis, of the USP Amodiaquine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of amodiaquine in the CU Sample solution (g/mL) F = molecular weight correction factor, 0.7656 Acceptance criteria: 93.0%107.0% AU AS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 342 nm Sample solution: Sample per Dissolution 711. Standard solution: USP Amodiaquine Hydrochloride RS in Medium Analysis Samples: Sample solution and Standard solution Determine the amount of C20H22ClN3O 2HCl 2H2O dissolved from UV absorbances on filtered portions of the solution under test, suitably diluted with water, if necessary, in comparison with a Standard solution having a known concentration of USP Amodiaquine Hydrochloride RS. Tolerances: An amount of C20H22ClN3O 2HCl 2H2O equivalent to NLT 75% (Q) of the labeled amount of C20H22ClN3O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amodiaquine Hydrochloride RS
solvent to evaporate, and examine the plate under shortwavelength UV light. Acceptance criteria: The chromatograms show principal spots at about the same RF value, and no secondary spot, if present in the chromatogram from the Sample solution, is more intense than the principal spot of Standard solution B. SPECIFIC TESTS WATER DETERMINATION, Method I 921: 7.0%9.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amodiaquine Hydrochloride RS
Amodiaquine Hydrochloride Tablets

(Comment on this Monograph)id=m4050=Amodiaquine Hydrochloride Tablets=A-Monos.pdf) DEFINITION Amodiaquine Hydrochloride Tablets contain an amount of amodiaquine hydrochloride (C20H22ClN3O 2HCl 2H2O) equivalent to NLT 93.0% and NMT 107.0% of the labeled amount of amodiaquine (C20H22ClN3O). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Powder one or more Tablets, and transfer a portion of the powder, equivalent to 50 mg of amodiaquine, to a 125-mL separator. Add 20 mL of water, and shake for 1 min. Add 25 mL of chloroform and 1 mL of ammonium hydroxide, shake for 2 min, and when settled, filter the chloroform extract through cotton that previously has been rinsed with chloroform, collecting the extract in a vessel suitable for evaporation. Evaporate the chloroform, and dry the residue at 105 for 1 h. B. ULTRAVIOLET ABSORPTION 197U Sample solution: Prepare as directed in the Assay. ASSAY PROCEDURE Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, nominally equivalent to 300 mg of amodiaquine, to a 250-mL beaker, add 100 mL of dilute hydrochloric acid (1 in 100), and heat on a steam bath for about 15 min with occasional stirring. Cool the mixture to room temperature, transfer to a 200-mL volumetric flask, add dilute hydrochloric acid (1 in 100) to volume. Pipet 10 mL of the clear supernatant into a 125-mL separator. Add 10 mL of dilute hydrochloric acid (1 in 100), and wash with 20 mL of chloroform, discarding the washing. Add 4.5 mL of 1 N sodium hydroxide, and extract with four 25-mL portions of chloroform. Extract the combined chloroform solutions with three 50-mL portions of dilute hydrochloric
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Official Monographs / Amoxapine 219

SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 177181 LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amoxapine RS
Amoxapine
(Comment on this Monograph)id=m4070=Amoxapine=AMonos.pdf)
C17H16ClN3O Dibenz[b,f][1,4]oxazepine, 2-chloro-11-(1-piperazinyl)-; 2-Chloro-11-(1-piperazinyl)dibenz[b,f][1,4]oxazepine [14028-44-5]. DEFINITION Amoxapine contains NLT 98.5% and NMT 101.0% of C17H16ClN3O, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K
313.78
Amoxapine Tablets
(Comment on this Monograph)id=m4080=Amoxapine Tablets=A-Monos.pdf) DEFINITION Amoxapine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of amoxapine (C17H16ClN3O). IDENTIFICATION INFRARED ABSORPTION 197K Sample: Triturate a quantity of finely ground Tablets, equivalent to 50 mg of amoxapine, with 10 mL of chloroform, and filter. Evaporate the filtrate on a steam bath to dryness (about 30 min). ASSAY PROCEDURE Solution A: 1.38 mg/mL of monobasic sodium phosphate in water Solution B: 113 mg/mL of tetramethylammonium chloride in water Mobile phase: Acetonitrile, 0.01 M monobasic sodium phosphate, 1 M tetramethylammonium chloride, and dilute phosphoric acid (1 in 5). (180:309:10:1). Standard stock solution: 1 mg/mL of USP Amoxapine RS in acetonitrile. Shake by mechanical means to dissolve, and then dilute with acetonitrile to volume. Standard solution: 0.1 mg/mL from Standard stock solution diluted with Mobile phase Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 50 mg of amoxapine, to a 50-mL volumetric flask. Add 40 mL of Mobile phase, and shake vigorously by mechanical means for 20 min. Dilute with Mobile phase to volume and filter. Pipet 5.0 mL of the filtrate into a 50-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1200 theoretical plates from the analyte peak Tailing factor: NMT 1.8 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C17H16ClN3O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100
ASSAY PROCEDURE Sample: 300 mg Analysis: Transfer the Sample to a 250-mL flask, dissolve in 50 mL of glacial acetic acid, and add 3 drops of crystal violet TS. Titrate with 0.1 N perchloric acid VS to an emerald green endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 15.69 mg of C17H16ClN3O. Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Standard solution A: 0.50 mg/mL of USP Amoxapine RS in chloroform Standard solution B: 0.25 mg/mL USP Amoxapine RS in chloroform; from Standard solution A Sample solution: 50 mg/mL of Amoxapine in chloroform Developing solvent system: Chloroform, methanol, and ammonium hydroxide (18:2:0.1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.2-mm layer of chromatographic silica gel mixture Application volume: 5 L Analysis Samples: Standard solution A, Standard solution B, and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, examine it under short-wavelength UV light, and compare the intensities of any secondary spots observed in the chromatogram of the Sample solution with those of the principal spots in the chromatogram of the Standard solutions. Acceptance criteria: No secondary spot from the chromatogram of the Sample solution is larger or more intense than the principal spot of Standard solution B (0.5%), and the sum of the intensities of the secondary spots of the Sample solution corresponds to NMT 1.0%.
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Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Diluent. [NOTEUse this solution within 6 h.] Sample solution: 1.2 mg/mL of Amoxicillin in Diluent. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C16H19N3O5S/mg taken: Result = (rU/rS) (CS/CU) P = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) CU = concentration of Sample solution (mg/mL) P = stated amoxicillin content of USP Amoxicillin RS (g/mg) Acceptance criteria: 9001050 g SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements DIMETHYLANILINE 223: Meets the requirement PH 791: 3.56.0 Sample solution: 2 mg/ml WATER DETERMINATION, Method I 921: 11.5%14.5% STERILITY TESTS 71: Where the label states that Amoxicillin is sterile, it meets the requirements when tested as directed in Test for Sterility of the Product to Be Examined, Direct Inoculation of the Culture Medium, except to use Fluid Thioglycollate Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the amoxicillin in each tube, to use SoybeanCasein Digest Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the amoxicillin in each tube, and to shake the tubes once daily. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Amoxicillin is sterile or Amoxicillin must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.25 Endotoxin Unit/mg of amoxicillin. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is intended for veterinary use only and that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. Label all other Amoxicillin to indicate that it is to be used in the manufacture of nonparenteral drugs only. USP REFERENCE STANDARDS 11 USP Amoxicillin RS USP Endotoxin RS rU rS CS
= amoxapine peak area from the Sample solution = amoxapine peak area from the Standard solution = concentration of USP Amoxapine RS in the Standard solution (mg/mL) = nominal concentration of amoxapine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Simulated gastric fluid (without enzyme); 900 mL. Apparatus 2: 50 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Standard solution: USP Amoxapine RS in Medium Spectrometric conditions Analytical wavelength: 294 nm Analysis: Determine the amount of C17H16ClN3O dissolved from UV absorbances of filtered portions of the Sample solution, suitably diluted with Medium, if necessary, in comparison with a Standard solution having a known concentration of USP Amoxapine RS. Acceptance criteria: NLT 80% (Q) of the labeled amount of C17H16ClN3O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amoxapine RS
Amoxicillin
(Comment on this Monograph)id=m4100=Amoxicillin=AMonos.pdf)
419.45 C16H19N3O5S 3H2O 4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[ [amino(4-hydroxyphenyl)acetyl]amino-3,3-dimethyl-7-oxo-, trihydrate [2S-[2,5,6(S*)]]-; (2S,5R,6R)-6-[(R)-(-)-2-Amino-2-(phydroxyphenyl)acetamido]-3,3-dimethyl-7-oxo-4-thia-1azabicyclo[3.2.0]heptane-2-carboxylic acid trihydrate [61336-70-7]. Anhydrous 365.41 [26787-78-0]. DEFINITION Amoxicillin contains NLT 900 g and NMT 1050 g of C16H19N3O5S per mg, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Diluent: 6.8 g/L of monobasic potassium phosphate in water, and adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1 Mobile phase: Acetonitrile and Diluent (1:24). Decrease the acetonitrile concentration to increase the retention time of amoxicillin.
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= amoxicillin peak response from the Standard solution CS = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) P = stated content of USP Amoxicillin RS (mg/mg) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701 Medium: Simulated gastric fluid being used instead of water Time: 30 min SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 7.5% rS
Amoxicillin Boluses
(Comment on this Monograph)id=m4105=Amoxicillin Boluses=A-Monos.pdf) DEFINITION Amoxicillin Boluses contain NLT 90.0% and NMT 110.0% of the labeled amount of amoxicillin (C16H19N3O5S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid. [NOTEUse within 10 min of preparation.] Sample solution: 4 mg/mL of amoxicillin, from powdered Boluses in 0.1 N hydrochloric acid Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine, and water (9:8:1:3) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, and dry with warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent, and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Diluent: 6.8 mg/mL of monobasic potassium phosphate in water. Adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1. Mobile phase: Acetonitrile and Diluent (1:24). Decrease the acetonitrile concentration to increase the retention time of amoxicillin. Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Diluent. [NOTEUse this solution within 6 h.] Sample solution: Weigh and finely powder NLT 5 Boluses. Transfer a portion of the powder, equivalent to 250 mg of amoxicillin, to a 250-mL volumetric flask, add Diluent to volume, and mix. Sonicate if necessary to ensure complete dissolution of the amoxicillin. Pass a portion of this solution through a filter of 1-m or finer porosity. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O5S in the portion of Boluses taken: Result = (rU/rS) (CS/CU) 100 P rU = amoxicillin peak response from the Sample solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label Boluses to indicate that they are for veterinary use only. USP REFERENCE STANDARDS 11 USP Amoxicillin RS
Amoxicillin Capsules
(Comment on this Monograph)id=m4110=Amoxicillin Capsules=A-Monos.pdf) DEFINITION Amoxicillin Capsules contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of amoxicillin (C16H19N3O5S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid [NOTEUse within 10 min after preparation.] Sample solution: Equivalent to 4 mg/mL of amoxicillin, from Capsule powder in 0.1 N hydrochloric acid Chromatographic system (see Chromatography 621, Thin-layer chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine, and water (9:8:1:3) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution When the solvent front has moved three-fourths of the length of the plate, remove the plate from the chamber, and dry with warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Diluent: Dissolve 6.8 g/L of monobasic potassium phosphate in water, and adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1. Mobile phase: Acetonitrile and Diluent (1:24). Decrease the acetonitrile concentration to increase the retention time of amoxicillin.
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USP 32
Meet the requirements NMT 14.5%
Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Diluent. [NOTE Use this solution within 6 h.] Sample solution: Remove, as completely as possible, the contents of NLT 20 Capsules, mix the combined contents, and transfer a quantity, equivalent to 200 mg of anhydrous amoxicillin, to a 200-mL volumetric flask, and add Diluent to volume. Sonicate if necessary to ensure complete dissolution. Pass a portion of this solution through a suitable filter having a 1-m or finer porosity, and use the filtrate. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O5S in the portion of Capsules taken: Result = (rU/rS) (CS/CU) P 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) CU = nominal concentration of amoxicillin in the Sample solution (mg/mL) P = stated amoxicillin content of USP Amoxicillin RS (mg/mL) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1 Medium: Water; 900 mL Apparatus 1: 100 rpm, for Capsules containing 250 mg Apparatus 2: 75 rpm, for Capsules containing 500 mg Time: 60 min Analytical wavelength: UV 272 nm Standard solution: USP Amoxicillin RS in Medium Sample solution: Sample per Dissolution 711; dilute with Medium to a concentration that is similar to Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C16H19N3O5S Test 2 (If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2.) Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 90 min Analytical wavelength: UV 272 nm Standard solution: USP Amoxicillin RS in Medium Sample solution: Sample per Dissolution 711; dilute with Medium to concentration that is similar to Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C16H19N3O5S rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: When more than one Dissolution test is given, the labeling states the Dissolution test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Amoxicillin RS
Amoxicillin Intramammary Infusion

(Comment on this Monograph)id=m4120=Amoxicillin Intramammary Infusion=A-Monos.pdf) DEFINITION Amoxicillin Intramammary Infusion is a suspension of Amoxicillin in a suitable vegetable oil vehicle. It contains NLT 90.0% and NMT 120.0% of the labeled amount of amoxicillin (C16H19N3O5S). It contains a suitable dispersing agent and preservative. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid. [NOTEUse within 10 min after preparation.] Sample solution: Transfer a quantity of Intramammary Infusion, equivalent to 60 mg of amoxicillin, to a 50-mL centrifuge tube. Add 25 mL of toluene, and centrifuge. Decant and discard the toluene. Wash the residue with four 25-mL portions of toluene, sonicating for 30 s after each addition of toluene. Dry the residue in a vacuum over silica gel. Add 15 mL of 0.1 N hydrochloric acid to the residue. Chromatographic system (see Chromatography 621, Thin-layer chromatography.) Adsorbent: 0.25-mm layer of chromatographic silica gel mixture (see Chromatography 621) Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine, water, and (9:8:1:3) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, and dry with warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Analysis: Proceed as directed for amoxicillin under AntibioticsMicrobial Assays 81. Expel the contents of 1 syringe of Intramammary Infusion into a high-speed glass blender jar containing 499.0 mL of Buffer No. 3 and 1.0 mL of polysorbate 80, and blend for 35 min. Allow to stand for 10 min, and dilute a measured volume of the aqueous phase quantitatively and stepwise with Buffer No. 3 to obtain
USP 32
a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%120.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1.0%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed disposable syringes. LABELING: Label it to indicate that it is intended for veterinary use only. USP REFERENCE STANDARDS 11 USP Amoxicillin RS

finer porosity, and use the filtrate as Sample solution A. [NOTEUse this solution within 6 h.] Sample solution B (where the label states the quantity of amoxicillin in a given volume of constituted suspension): Constitute Amoxicillin for Injectable Suspension as directed in the labeling. Quantitatively dilute a measured volume of the constituted suspension with Diluent to obtain a solution containing 1 mg/mL of anhydrous amoxicillin. Pass a portion of this solution through a suitable filter of 1-m or finer porosity, and use the filtrate as Sample solution B. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solutions Calculate the percentage of C16H19N3O5S in the container, or in the portion of constituted Amoxicillin for Injectable Suspension taken: Result = (rU/rS) (Cs/CU) P 100 = amoxicillin peak response of the Sample solution = amoxicillin peak response of the Standard solution CS = concentration of USP Amoxicillin RS in the Standard solution CU = nominal concentration, in mg of anhydrous amoxicillin/mL, of Sample solution A or of Sample solution B on the basis of the labeled quantity in the container or in the portion of constituted suspension taken, respectively, and the extent of dilution P = stated content of USP Amoxicillin RS (mg/mg) Acceptance criteria: 90.0%120.0% SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed in Test for Sterility of the Product to Be Examined, Direct Inoculation of the Culture Medium, except to use Fluid Thioglycollate Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the amoxicillin in each tube, to use SoybeanCasein Digest Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the amoxicillin in each tube, and to shake the tubes once daily. PH 791: 5.07.0, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: 11.0%14.0% BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.25 Endotoxin Unit/mg of amoxicillin. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in Injections 1, Containers for Sterile Solids. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Amoxicillin RS rU rS
Amoxicillin for Injectable Suspension

(Comment on this Monograph)id=m4129=Amoxicillin for Injectable Suspension=A-Monos.pdf) DEFINITION Amoxicillin for Injectable Suspension is a sterile mixture of Amoxicillin and one or more suitable buffers, preservatives, stabilizers, and suspending agents. It contains NLT 90.0% and NMT 120.0% of the labeled amount of amoxicillin (C16H19N3O5S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid. [NOTEUse within 10 min after preparation.] Sample solution: Equivalent of 4 mg/mL of amoxicillin, from Amoxicillin for Injectable Suspension, in 0.1 N hydrochloric acid. Allow to stand for 5 min before use. Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine, and water (9:8:1:3) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, and dry with warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent, and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot obtained from the Sample solution corresponds to that obtained from the Standard solution. ASSAY PROCEDURE Diluent: 6.8 mg/mL of monobasic potassium phosphate in water. Adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1. Mobile phase: Acetonitrile and Diluent (1:24). Decrease the acetonitrile concentration to increase the retention time of amoxicillin. Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Diluent. [NOTEUse this solution within 6 h.] Sample solution A (where it is represented as being in a single-dose container): Constitute Amoxicillin for Injectable Suspension as directed in the labeling. Withdraw all of the withdrawable contents, using a hypodermic needle and syringe, and quantitatively dilute with Diluent to obtain a solution containing 1 mg/mL of anhydrous amoxicillin. Pass a portion of this solution through a suitable filter of 1-m or
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in multiple-dose containers equipped with a suitable dosing pump. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Amoxicillin RS
USP Endotoxin RS
Amoxicillin Oral Suspension

(Comment on this Monograph)id=m4130=Amoxicillin Oral Suspension=A-Monos.pdf) DEFINITION Amoxicillin Oral Suspension is a suspension of Amoxicillin in Soybean Oil. It contains NLT 90.0% and NMT 120.0% of the labeled amount of amoxicillin (C16H19N3O5S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid. [NOTEUse within 10 min after preparation.] Sample solution: Shake a portion of Oral Suspension with a mixture of acetone and 0.1 N hydrochloric acid (4:1) to obtain a solution containing 1 mg/mL of amoxicillin. Chromatographic system (See Chromatography 621, Thin-layer chromatography.) Adsorbent: 0.25-mm layer of chromatographic silica gel mixture (See Chromatography 621.) Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine and water, (9:8:1:3) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Sample solution and Standard solution When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, and dry with warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Standard solution: Prepare as directed for Standard Preparation under Iodometric AssayAntibiotics 425, using USP Amoxicillin RS. Sample solution: Using the dosing pump, deliver a number of doses of Oral Suspension, equivalent to 250 mg of amoxicillin, to a separator containing 100 mL of hexanes, and shake vigorously. Add 140 mL of water, and shake for 5 min. Allow the layers to separate, and drain the lower, aqueous layer into a 250-mL volumetric flask. Repeat the extraction with two 50-mL portions of water. Combine the aqueous extracts in the volumetric flask, dilute with water to volume. Analysis: Proceed with Oral Suspension as directed for Procedure under Iodometric AssayAntibiotics 425, using USP Amoxicillin RS. Calculate the percentage of C16H19N3O5S in each dose of Oral Suspension taken: (250/N)(F/2000)(B I) = the number of doses taken, and the other terms are as defined therein. Acceptance criteria: 90.0%120.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 2.0%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel. N
Amoxicillin for Oral Suspension

(Comment on this Monograph)id=m4140=Amoxicillin for Oral Suspension=A-Monos.pdf) DEFINITION Amoxicillin for Oral Suspension contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of amoxicillin (C16H19N3O5S). It contains one or more suitable buffers, colors, flavors, preservatives, stabilizers, sweeteners, and suspending agents. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid. [NOTEUse within 10 min after preparation.] Sample solution: Equivalent of 4 mg/mL of amoxicillin, from Oral Suspension in 0.1 N hydrochloric acid. [NOTE Allow the solution to stand for 5 min before use.] Chromatographic system (see Chromatography 621, System Suitability.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine, and water (9:1:8:3) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution When the solvent front has moved three-fourths of the length of the plate, remove the plate from the chamber, and dry with warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Diluent: Dissolve 6.8 g/L of monobasic potassium phosphate in water, and adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1. Mobile phase: Acetonitrile and Diluent (1:24). Decrease the acetonitrile concentration to increase the retention time of amoxicillin. Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Diluent. [NOTEUse this solution within 6 h.] Sample solution: Dilute a measured volume of Amoxicillin for Oral Suspension, constituted as directed in the labeling, freshly mixed and free from air bubbles, quantitatively and stepwise in Diluent to obtain a solution containing nominally 1 mg/mL of anhydrous amoxicillin. Filter a portion of this solution through a suitable filter of 1-m or finer porosity, and use the filtrate as the Sample solution. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O5S in each mL of the constituted Amoxicillin for Oral Suspension taken: Result = (rU/rS) (CS/CU) P 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) = nominal concentration of anhydrous amoxicillin CU in the Sample solution (mg/mL) P = stated amoxicillin content of USP Amoxicillin RS (g/mg) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 For solids packaged in single-unit containers: Meets the requirements DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 5.07.5, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 3.0%, except where it is labeled as containing 80 mg of amoxicillin per mL after constitution, the limit is NMT 4.0%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Amoxicillin RS rU rS CS

Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution Dry the plate with the aid of a current of warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent, and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Diluent: 6.8 g/L of monobasic potassium phosphate in water, and adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1 Mobile phase: Acetonitrile and Diluent (1:24). Decrease the acetonitrile concentration to increase the retention time of amoxicillin. Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Diluent. [NOTEUse this solution within 6 h.] Sample solution: Place NLT 5 Tablets in a high-speed glass blender jar containing Diluent sufficient to yield a concentration of 1 mg of anhydrous amoxicillin/mL, blend for 4 1 min, allow to stand for 5 min, and centrifuge a portion of the mixture. [NOTEWhere the volume of Diluent required would exceed 500 mL, place 5 Tablets in a volumetric flask of such capacity that when finally diluted to volume, a concentration of 1 mg of anhydrous amoxicillin per mL would be obtained. Add a volume of Diluent equivalent to three-fourths of the capacity of the volumetric flask, and sonicate for 5 min. Dilute with Diluent to volume, add a magnetic stirring bar, and stir for 30 min. Centrifuge a portion of this solution.] Pass a portion of the clear supernatant through a suitable filter having a 1-m or finer porosity, and use the filtrate. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19 N3O5S in each Tablet taken: Result = (rU/rS) (CS/CU) P 100 rU rS CS CU P = peak response of amoxicillin from the Sample solution = peak response of amoxicillin from the Standard solution = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) = nominal concentration of amoxicillin in the Sample solution (mg/mL) = stated content of USP Amoxicillin RS (mg/mg)
Amoxicillin Tablets
(Comment on this Monograph)id=m4170=Amoxicillin Tablets=A-Monos.pdf) DEFINITION Amoxicillin Tablets contain NLT 90.0% and NMT 120.0% of the labeled amount of amoxicillin (C16H19N3O5S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 4 mg/mL, from powdered Tablets in 0.1 N hydrochloric acid. [NOTEUse within 10 min after preparation.] Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine, and water (9:8:1:3)
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90.0%120.0%
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For veterinary products: Proceed as directed above, except to use Apparatus 2 at 100 rpm. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label chewable Tablets to indicate that they are to be chewed before swallowing. Tablets intended solely for veterinary use are so labeled. USP REFERENCE STANDARDS 11 USP Amoxicillin RS
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 30 min Determine the amount of C16H19N3O5S dissolved by employing the following method. pH 5.0 Buffer: 27.2 g of monobasic potassium phosphate in 3 L of water, adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1, and dilute with water to obtain 4 L of solution Mobile phase: Acetonitrile and pH 5.0 Buffer (1:39), and pass through a filter having a 0.5-m or finer porosity Standard solution: 0.05 mg/mL of USP Amoxicillin RS in pH 5.0 Buffer. [NOTEUse this solution within 6 h.] Sample solution: Pass a portion of the sample through a filter having a 0.5-m or finer porosity. Quantitatively dilute a volume of the filtrate with water to obtain a concentration of 0.045 mg/mL of amoxicillin. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column Analytical: 3.9-mm 30-cm; packing L1 Guard: 2-mm 2-cm; packing L2 Temperature: Analytical column is maintained at a constant temperature of 40 1 Flow rate: 0.7 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O5S dissolved by the formula: Result = (rU/rS) (CS D V P (100/L) = peak response of amoxicillin from the Sample solution = peak response of amoxicillin from the Standard rS solution CS = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) V = volume of the dissolution medium, 900 mL D = dilution factor for the Sample solution P = stated content of USP Amoxicillin RS (mg/mg) L = label claim (mg/tablet) Time: 30 min Tolerances: NLT 75% (Q) of the labeled amount of C16H19N3O5S For products labeled as chewable tablets: Proceed as directed above. For chewable tablets labeled to contain 200 mg or 400 mg Time: 20 min Tolerances: NLT 70% (Q) of the labeled amount of C16H19N3O5S For chewable tablets labeled to contain 125 mg or 250 mg Time: 90 min Tolerances: NLT 70% (Q) of the labeled amount of C16H19N3O5S rU
Amoxicillin Tablets for Oral Suspension

(Comment on this Monograph)id=m4173=Amoxicillin Tablets for Oral Suspension=A-Monos.pdf) DEFINITION Amoxicillin Tablets for Oral Suspension contain NLT 90.0% and NMT 110.0% of the labeled amount of amoxicillin (C16H19N3O5S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: An aqueous dispersion of Amoxicillin Tablets for Oral Suspension in 0.1 N hydrochloric acid containing 4 mg/mL of amoxicillin. Use within 10 min of preparation. Standard solution: 4 mg/mL of USP Amoxicillin RS in 0.1 N hydrochloric acid Chromatographic system (see Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Methanol, chloroform, pyridine, and water (9:8:1:3) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution Dry the plate with the aid of a current of warm air for 10 min. Locate the spots on the plate by spraying lightly with Spray reagent, and dry at 110 for 15 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Diluent: 6.8 g/L of monobasic potassium phosphate in water, and adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1 Mobile phase: Acetonitrile and Diluent (1:24). Decrease the acetonitrile concentration to increase the retention time of amoxicillin. Standard solution: 1.2 mg/mL of USP Amoxicillin RS in Diluent. [NOTEUse this solution within 6 h.] Sample solution: Prepare a dispersion of 20 Tablets for Oral Suspension using an accurately measured volume of water. Quantitatively dilute a portion of the dispersion with Diluent to obtain a solution containing 1.2 mg/mL of amoxicillin. Pass a portion of the solution through a filter having a 1-m or finer porosity, and use the filtrate. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O5S in each Amoxicillin Tablet for Oral Suspension taken: Result = (rU/rS) (CS/CU) P 100 = amoxicillin peak response from the Sample solution = amoxicillin peak response from the Standard rS solution = concentration of USP Amoxicillin RS in the CS Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) P = stated content of USP Amoxicillin RS (mg/mg) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701 Medium: Water at 20 5 Time: 3 min DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 30 min Determine the amount of C16H19N3O5S dissolved by employing the following method. pH 5.0 buffer: 27.2 g of monobasic potassium phosphate in 3 L of water, adjust with a 45% (w/w) solution of potassium hydroxide to a pH of 5.0 0.1, dilute with water to obtain 4 L of solution Mobile phase: Acetonitrile and pH 5.0 buffer (1:39), and pass through a filter having a 0.5-m or finer porosity Standard solution: 0.05 mg/mL of USP Amoxicillin RS in pH 5.0 buffer. [NOTEUse this solution within 6 h.] Sample solution: Pass a portion of the sample through a filter having a 0.5-m or finer porosity. Quantitatively dilute an accurately measured volume of the filtrate with water to obtain a concentration of 0.045 mg/mL of amoxicillin. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column Analytical column: 3.9-mm 30-cm; packing L1 Guard column: 2-mm 2-cm; packing L2 Temperature: Analytical column is maintained at a constant temperature of 40 1 rU

Flow rate: 0.7 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 1.12.8 Column efficiency: NLT 1700 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 1.5% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C16H19N3O5S dissolved: Result = (rU/rS) (CS D V P (100/L) = amoxicillin peak responses from the Sample solution = amoxicillin peak responses from the Standard rS solution CS = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) V = volume of medium, 900 D = dilution factor required to prepare the Sample solution P = stated content of USP Amoxicillin RS (g/mg) L = label claim (mg/tablet) Tolerances: NLT 80% (Q) of the labeled amount of C16H19N3O5S UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU SPECIFIC TESTS DISPERSION FINENESS: Place 2 Tablets for Oral Suspension in 100 mL of water, and stir until completely dispersed. A smooth dispersion is obtained that passes through a No. 25 sieve. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Amoxicillin RS
Amoxicillin and Clavulanate Potassium for Oral Suspension

(Comment on this Monograph)id=m4190=Amoxicillin and Clavulanate Potassium for Oral Suspension=A-Monos.pdf) DEFINITION Amoxicillin and Clavulanate Potassium for Oral Suspension contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of amoxicillin (C16H19N3O5S) and the equivalent of NLT 90.0% and NMT 125.0% of the labeled amount of clavulanic acid (C8H9NO5). It contains one or more suitable buffers, colors, flavors, preservatives, stabilizers, sweeteners, and suspending agents. IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay.
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PERFORMANCE TESTS DELIVERABLE VOLUME 698 For powder packaged in multiple-unit containers: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905 For powder packaged in single-unit containers: Meets the requirements SPECIFIC TESTS PH 791: 3.86.6, in the suspension constituted as directed in the labeling, the test being performed immediately after constitution. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Amoxicillin RS USP Clavulanate Lithium RS
ASSAY PROCEDURE Buffer: 7.8 g of monobasic sodium phosphate in 900 mL of water, adjust with phosphoric acid or 10 N sodium hydroxide to a pH of 4.4 0.1, and dilute with water to 1000 mL. Mobile phase: Methanol and Buffer (1:19), and pass through a filter having a 0.5-m or finer porosity. Standard solution: 0.5 mg/mL of USP Amoxicillin RS and 0.2 mg/mL of USP Clavulanate Lithium RS in water Sample solution: Dilute an accurately measured volume of Amoxicillin and Clavulanic Acid for Oral Suspension, constituted as directed in the labeling, quantitatively with water to obtain a solution containing 0.5 mg/mL of amoxicillin. Stir by mechanical means for 10 min, and filter. Use the filtrate as the Sample solution within 1 h of the dilution of the suspension. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4-mm 30-cm; 3- to 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for clavulanic acid and amoxicillin are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.5 between the amoxicillin and clavulanic acid peaks Column efficiency: NLT 550 theoretical plates from each analyte peak Tailing factor: NMT 1.5 for each analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O5S in each mL of the constituted Amoxicillin and Clavulanate Potassium for Oral Suspension taken: Result = (rU/rS) (CS/CU) P 100 rU = amoxicillin peak response from the Sample solution rS = amoxicillin peak response from the Standard solution CS = concentration of USP Amoxicillin RS in the Standard solution (mg/mL) CU = nominal concentration of amoxicillin in the Sample solution (mg/mL) P = stated content of USP Amoxicillin RS (mg/mg) Calculate the percentage of C8H9NO5 in each mL of the constituted Amoxicillin and Clavulanate Potassium for Oral Suspension taken: Result = (rU/rS) (CS/CU) P 100 = clavulanic acid peak response from the Sample solution = clavulanic acid peak response from the Standard rS solution = concentration of USP Clavulanate Lithium RS in CS the Standard solution (mg/mL) = nominal concentration of clavulanate in the CU Sample solution (mg/mL) P = stated content of USP Clavulanate Potassium RS (mg/mg) Acceptance criteria: 90.0%120.0% of the labeled amount of C16H19N3O5S and 90.0%125.0% of the labeled amount of C8H9NO5 rU
Amoxicillin and Clavulanate Potassium Tablets

(Comment on this Monograph)id=m4195=Amoxicillin and Clavulanate Potassium Tablets=A-Monos.pdf) DEFINITION Amoxicillin and Clavulanate Potassium Tablets contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amounts of amoxicillin (C16H19N3O5S) and clavulanic acid (C8H9NO5). IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: 7.8 g of monobasic sodium phosphate in 900 mL of water, adjust with phosphoric acid or 10 N sodium hydroxide to a pH of 4.4 0.1, and dilute with water to 1000 mL Mobile phase: Methanol and Buffer (1:19), and pass through a filter having a 0.5-m or finer porosity Standard solution: 0.5 mg/mL of USP Amoxicillin RS and 0.2 mg/mL of USP Clavulanate Lithium RS in water Sample stock solution: Dissolve NLT 10 Tablets in water with the aid of mechanical stirring, transfer to a suitable volumetric flask, and dilute with water to volume. Filter a portion of this solution, discarding the first 10 mL of the filtrate. Sample solution: Dilute an accurately measured volume of the Sample stock solution filtrate with water to obtain a solution containing 0.5 mg/mL of amoxicillin. Use this Sample solution within 1 h. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4-mm 30-cm; 3- to 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for clavulanic acid and amoxicillin are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.5 between the amoxicillin and clavulanic acid peaks
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Column efficiency: NLT 550 theoretical plates from each analyte peak Tailing factor: NMT 1.5 for each analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O5S in each Tablet taken: Result = (rU/rS) (CS/CU) P 100 = amoxicillin peak response from the Sample solution = amoxicillin peak response from the Standard rS solution = concentration of USP Amoxicillin RS in the CS Standard solution (mg/mL) CU = nominal concentration of amoxicillin in the Sample solution (mg/mL) P = stated content of USP Amoxicillin RS (mg/mg) Calculate the percentage of C8H9NO5 in each Tablet taken: Result = (rU/rS) (CS/CU) P 100 = clavulanic acid peak response from the Sample solution = clavulanic acid peak response from the Standard rS solution = concentration of USP Clavulanate Lithium RS in CS the Standard solution (mg/mL) CU = nominal concentration of clavulanate in the Sample solution (mg/mL) P = stated content of USP Clavulanate Lithium RS (mg/mg) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISINTEGRATION 701: Tablets labeled for veterinary use only; 30 min, simulated gastric fluid TS being substituted for water in the test. DISSOLUTION 711 [NOTETablets labeled for veterinary use only are exempt from this requirement.] Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 30 min; or 45 min where the Tablets are labeled as chewable. Analysis: Determine the amount of C16H19N3O5S and C8H9NO5 dissolved, employing the Analysis set forth in the Assay, making any necessary volumetric adjustments. Tolerances: NLT 85% (Q) of the labeled amount of C16H19N3O5S and NLT 80% (Q) of the labeled amount of C8H9NO5 For tablets labeled as chewable: NLT 80% (Q) of the labeled amounts of C16H19N3O5S and C8H9NO5 is dissolved in 45 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to amoxicillin and for Content Uniformity with respect to clavulanic acid. SPECIFIC TESTS WATER DETERMINATION, Method I 921 NMT 7.5% where the labeled amount of amoxicillin in each Tablet is 250 mg or less; NMT 10.0% where the labeled amount of amoxicillin in each Tablet is more than 250 mg but less than or equal to 500 mg; NMT 11.0% where the labeled amount of amoxicillin in each Tablet is more than 500 mg. Where Tablets are labeled as chewable NMT 6.0% where the labeled amount of amoxicillin in each Tablet is 125 mg or less; rU rU
Official Monographs / Amphetamine 229

NMT 8.0% where the labeled amount of amoxicillin in each Tablet is more than 125 mg; Where the Tablets are labeled for veterinary use only NMT 10.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label chewable Tablets to include the word chewable in juxtaposition to the official name. The labeling indicates that chewable Tablets may be chewed before being swallowed or may be swallowed whole. Tablets intended for veterinary use only are so labeled. USP REFERENCE STANDARDS 11 USP Amoxicillin RS USP Clavulanate Lithium RS
Amphetamine Sulfate
(Comment on this Monograph)id=m4260=Amphetamine Sulfate=A-Monos.pdf)
(C9H13N)2 H2SO4 Benzeneethanamine, -methyl-, sulfate (2:1), ()-; ()--Methylphenethylamine sulfate (2:1) [60-13-9].
368.49
DEFINITION Amphetamine Sulfate, dried at 105 for 2 h, contains NLT 98.0% and NMT 102.0% of (C9H13N)2 H2SO4. IDENTIFICATION A. MELTING RANGE OR TEMPERATURE, Class I 741 Sample solution: 20 mg/mL Amphetamine Sulfate Analysis: To 5 mL of Sample solution add 5 mL of 1 N sodium hydroxide, cool to 10, add 1 mL of a mixture of absolute ether and benzoyl chloride (2:1), insert the stopper, and shake for 3 min. Filter the precipitate, wash with 10 mL of cold water, and recrystallize from diluted alcohol. Acceptance criteria: The crystals of the benzoyl derivative of amphetamine so obtained, after drying at 80 for 3 h, melt between 131 and 135. B. IDENTIFICATION TESTSGENERAL, Sulfate 191: Meets the requirements of the tests Sample solution: 100 mg/mL ASSAY PROCEDURE Sample: 500 mg of amphetamine Analysis: Dissolve Sample in 50 mL of glacial acetic acid, and titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 36.85 mg of (C9H13N)2 H2SO4. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE 1 Buffer solution: Dissolve 2.16 g of sodium 1octanesulfonate in 1000 mL of water. Add 1.0 mL of triethylamine, and adjust with phosphoric acid to pH of 2.5.
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water for 30 min, and filter into a small flask. To the filtrate add 3 mL of 1 N sodium hydroxide. Cool to 10 to 15, add 1 mL of a mixture of 1 volume of benzoyl chloride and 2 volumes of absolute ether, insert the stopper, and shake well for 3 min. Filter the precipitate, wash with 15 mL of cold water, and recrystallize twice from diluted alcohol. Acceptance criteria: The crystals of the benzoyl derivative of amphetamine so obtained, after drying at 80 for 2 h, melt between 131 and 135. ASSAY PROCEDURE Standard solution: Prepare as directed under Amphetamine Assay 331. Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 5 mg of amphetamine sulfate, to a 100-mL beaker, add 2 mL of hydrochloric acid solution (1 in 100), swirl gently to wet the powder thoroughly, warm on a steam bath for 1 min, with occasional gentle swirling, and cool. Add 3 g of purified siliceous earth, and mix until a fluffy mixture is obtained. Analysis: Proceed as directed under Amphetamine Assay 331. Calculate the quantity, in mg, of (C9H13N)2 H2SO4 in the portion of Tablets taken: Result = 0.01 C [(AU257 AU280)/(AS257 AS280)] = concentrationof USP Dextroamphetamine Sulfate RS in the Standard solution (g/mL) Acceptance criteria: 93.0%107.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 500 mL Apparatus 1: 100 rpm Time: 45 min Standard solution: USP Dextroamphetamine Sulfate RS in Medium Mobile phase: 1.1 g of sodium 1-heptanesulfonate in 575 mL of water. Add 25 mL of dilute glacial acetic acid (14 in 100) and 400 mL of methanol. Adjust by the dropwise addition of glacial acetic acid to a pH of 3.3 0.1, if necessary, filter, and degas the solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 500 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Sample: Filtered portion of the solution under test Calculate the percentage of (C9H13N)2 H2SO4 dissolved in comparison with a Standard solution having a known concentration of USP Dextroamphetamine Sulfate RS and similarly chromatographed. Acceptance criteria: NLT 75% (Q) is dissolved UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dextroamphetamine Sulfate RS C
Mobile phase: Acetonitrile, methanol, and Buffer solution (37:19:144) Diluent: 3.12 mL of phosphoric acid to 1000 mL with water Standard stock solution: 0.3 mg/mL of USP Dextroamphetamine Sulfate RS in Diluent Standard solution: 0.003 mg/mL of dextroamphetamine from Standard stock solution diluted with Diluent Sample solution: 0.3 mg/mL of Amphetamine Sulfate in Diluent. [NOTESonicate for 5 min and then dilute with Diluent to volume.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 215 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: Standard stock solution and Sample solution Suitability requirements Resolution: NLT 1.5, between amphetamine and any adjacent peak, if any, Sample solution Tailing factor: NMT 2.0, Standard stock solution Relative standard deviation: NMT 2.0%, Standard stock solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of each impurity in the portion of Amphetamine Sulfate taken: Result = (rI/rS) (CS/CU) 100 = peak response for each impurity from the Sample solution = peak response for amphetamine from the rS Standard solution = concentration of USP Dextroamphetamine CS Sulfate RS in the Standard solution (mg/mL) = concentration of Amphetamine Sulfate in the CU Sample solution (mg/mL) Acceptance criteria Individual impurities: NMT 0.1% Total impurities: NMT 0.5% PROCEDURE 2: DEXTROAMPHETAMINE: A solution (20 mg/mL) is optically inactive rI SPECIFIC TESTS LOSS ON DRYING 731: 1.0% of its weight. Dry it at 105 for 2 h: it loses NMT
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dextroamphetamine Sulfate RS
Amphetamine Sulfate Tablets

(Comment on this Monograph)id=m4290=Amphetamine Sulfate Tablets=A-Monos.pdf) DEFINITION Amphetamine Sulfate Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of (C9H13N)2 H2SO4. IDENTIFICATION MELTING RANGE OR TEMPERATURE, Class I 741 Sample solution: Macerate a quantity of powdered Tablets, equivalent to 50 mg of amphetamine sulfate, with 10 mL of
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Official Monographs / Amphotericin 231

a 50-mL volumetric flask, and dilute with methanol to volume. Transfer 4.0 mL of this solution to a 50-mL volumetric flask, and dilute with methanol to volume. [NOTEPrepare this solution fresh daily.] Spectrometric conditions Cell: 1 cm Measurement wavelength: 304 nm and 282 nm Blank: 16 mg/mL of dimethyl sulfoxide in methanol Analysis: Concomitantly determine the absorbances of the Nystatin standard solution and Amphotericin B standard solution and the Sample solution against the Blank. Calculate the percentage of amphotericin A taken:
Amphotericin B
(Comment on this Monograph)id=m4340=Amphotericin B=AMonos.pdf)
924.08 C47H73NO17 Amphotericin B; Result = 25 WN[(AB282 AU304) (AB304 AU282)]/[(AB282 AN304) 14,39-Dioxabicyclo[33.3.1]nonatriaconta-19,21,23,25,27,29,31 (AB304 AN282)] WU heptaene-36-carboxylic acid, 33-[(3-amino-3,6-dideoxy--Dmannopyranosyl)oxy]-1,3,5,6,9,11,17,37WN = weight of USP Nystatin RS taken (mg) octahydroxy-15,16,18-trimethyl-13-oxo-, AB282 = absorbance of the Amphotericin B standard (1R,3S,5R,6R,9R,11R,15S,16R,17R,18S, solution at 282 nm 19E,21E,23E,25E,27E,29E,31E,33R,35S,36R,37S)-; AU304 = absorbance of the Sample solution at 304 nm [1RAB304 = absorbance of the Amphotericin B standard (1R*,3S*,5R*,6R*,9R*,11R*,15S*,16R*,17R*,18S*,19E,21E,23E,25E,27E,29E,31E,33R*,35S*,36R*,37S*)]-33solution at 304 nm [(3-Amino-3,6-dideoxy--DAU282 = absorbance of the Sample solution at 282 nm mannopyranosyl)oxy]-1,3,5,6,9,11,17,37AN304 = absorbance of the Nystatin standard solution at octahydroxy-15,16,18-trimethyl-13-oxo-14,39-dioxabicyclo 304 nm [33.3.1]nonatriaconta-19,21,23,25,27,29,31-heptaene-36AN282 = absorbance of the Nystatin standard solution at carboxylic acid [1397-89-3]. 282 nm WU = weight of the Amphotericin B taken (mg) DEFINITION Acceptance criteria: NMT 5% Amphotericin B has a potency of NLT 750 g of C47H73NO17/ [NOTEAmphotericin B intended for use in preparing mg, calculated on the dried basis. dermatological creams, lotions, ointments, oral IDENTIFICATION suspensions, and capsules contains NMT 15% of ULTRAVIOLET ABSORPTION 197U amphotericin A, calculated on the dried basis.] Analytical wavelength I: 240320 nm SPECIFIC TESTS Solution I: Prepare as directed for Sample solution in the LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered Limit of amphotericin A, and compare its absorbance to that bottle in vacuum at a pressure not exceeding 5 mm of of the Amphotericin B standard solution. An extra peak may mercury at 60 for 3 h: it loses NMT 5.0% of its weight. occur at 304 nm in the spectrum of this solution. Analytical wavelength II: 320400 nm ADDITIONAL REQUIREMENTS Solution II: Prepare as directed for Sample solution in the PACKAGING AND STORAGE: Preserve in tight, light-resistant Limit of amphotericin A, and then dilute with 9 volumes of containers, and store in a cold place. methanol. Compare its absorbance to that of a similar LABELING: Label it to state whether it is intended for use in dilution of the Amphotericin B standard solution. preparing dermatological and oral dosage forms or parenteral dosage forms. ASSAY USP REFERENCE STANDARDS 11 PROCEDURE USP Amphotericin B RS Proceed with Amphotericin B as directed under Antibiotics USP Nystatin RS Microbial Assays 81. Acceptance criteria: NLT 750 g of C47H73NO17 per mg IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.5%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid [NOTEAmphotericin B intended for use in preparing dermatological creams, lotions, ointments, oral suspensions, and capsules, yields NMT 3.0%.] Organic Impurities PROCEDURE: LIMIT OF AMPHOTERICIN A Sample solution: Dissolve 50 mg of Amphotericin B in 10.0 mL of dimethyl sulfoxide in a 50-mL volumetric flask, and dilute with methanol to volume. Transfer 4.0 mL of this solution to a 50-mL volumetric flask, and dilute with methanol to volume. Nystatin standard solution: Dissolve 20 mg of USP Nystatin RS in 40.0 mL of dimethyl sulfoxide in a 200-mL volumetric flask, and dilute with methanol to volume. Transfer 4.0 mL of this solution to a 50-mL volumetric flask, and dilute with methanol to volume. Amphotericin B standard solution: Dissolve 50 mg of USP Amphotericin B RS in 10.0 mL of dimethyl sulfoxide in
Amphotericin B Cream
(Comment on this Monograph)id=m4350=Amphotericin B Cream=A-Monos.pdf) DEFINITION Amphotericin B Cream contains NLT 90.0% and NMT 125.0% of the labeled amount of Amphotericin B. ASSAY PROCEDURE Sample stock solution: Blend a suitable portion of Cream in a high-speed blender with a sufficient volume of dimethyl sulfoxide to give a convenient concentration. Quantitatively dilute a volume of this solution with dimethyl sulfoxide to obtain a concentration of 20 g/mL of amphotericin B. Sample solution: Quantitatively dilute a volume of Sample stock solution with Buffer No. 10 to obtain a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed for amphotericin B under AntibioticsMicrobial Assays 81.
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90.0%125.0% the solution should be protected from light during administration. USP REFERENCE STANDARDS 11 USP Amphotericin B RS USP Endotoxin RS
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PERFORMANCE TESTS MINIMUM FILL 755: It meets the requirements. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or other well-closed containers. USP REFERENCE STANDARDS 11 USP Amphotericin B RS
Amphotericin B Lotion
(Comment on this Monograph)id=m4370=Amphotericin B Lotion=A-Monos.pdf) DEFINITION Amphotericin B Lotion contains NLT 90.0% and NMT 125.0% of the labeled amount of amphotericin B. ASSAY PROCEDURE Sample stock solution: Quantitatively dissolve a suitable volume of Lotion in sufficient dimethyl sulfoxide to give a convenient concentration. Quantitatively dilute an accurately measured volume of this solution with dimethyl sulfoxide to obtain a concentration of 20 g/mL of amphotericin B. Sample solution: Quantitatively dilute an accurately measured volume of the Sample stock solution with Buffer No. 10 to obtain a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard. Procedure: Proceed as directed for amphotericin B under AntibioticsMicrobial Assays 81. Acceptance criteria: 90.0%125.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS PH 791: 5.07.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Amphotericin B RS
Amphotericin B for Injection

(Comment on this Monograph)id=m4360=Amphotericin B for Injection=A-Monos.pdf) DEFINITION Amphotericin B for Injection is a sterile complex of Amphotericin B, deoxycholate sodium, and one or more suitable buffers. It contains NLT 90.0% and NMT 120.0% of the labeled amount of C47H73NO17. ASSAY PROCEDURE Sample solution A (where it is packaged as a single-dose container): Constitute Amphotericin B for Injection as directed in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute quantitatively and stepwise with dimethyl sulfoxide to obtain a solution containing 20 g/mL of amphotericin B. Sample solution B (where the labeling states the quantity of amphotericin B in a given volume of constituted solution): Constitute Amphotericin B for Injection as directed in the labeling. Withdraw a volume of the resultant solution, using a suitable hypodermic needle and syringe, and dilute quantitatively and stepwise with dimethyl sulfoxide to obtain a solution containing 20 g/mL of amphotericin B. Analysis: Proceed as directed for amphotericin B under AntibioticsMicrobial Assays 81, using a volume of Sample solution diluted quantitatively and stepwise with Buffer No. 10 to obtain a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%120.0% SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed in Test for Sterility of the Product to be Examined, Membrane Filtration, 50 mg from each container being tested. PH 791: 7.28.0 Sample solution: 10 mg/mL of amphotericin B LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 8.0% of its weight. OTHER REQUIREMENTS: It meets the requirements for Uniformity of Dosage Units 905 and for InjectionsLabels and Labeling 1. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.0 USP Endotoxin Units/mg of amphotericin B. For products used or labeled for intrathecal injection, it contains NMT 0.9 USP Endotoxin Unit/mg of amphotericin B. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1, in a refrigerator and protected from light. LABELING: Label it to indicate that it is intended for use by intravenous infusion to hospitalized patients only, and that
Amphotericin B Ointment
(Comment on this Monograph)id=m4380=Amphotericin B Ointment=A-Monos.pdf) DEFINITION Amphotericin B Ointment is Amphotericin B in a suitable ointment base. It contains NLT 90.0% and NMT 125.0% of the labeled amount of amphotericin B. ASSAY PROCEDURE Sample stock solution: Mix a portion of Ointment, equivalent to 30 mg of amphotericin B, with 10.0 mL of ether in a suitable glass-stoppered conical flask and allow to stand, with intermittent shaking, for 1 h. Add 20.0 mL of dimethyl sulfoxide and shake by mechanical means for 10 min. Dilute quantitatively and stepwise with dimethyl sulfoxide to a concentration of 20 g/mL. Sample solution: Quantitatively dilute an accurately measured volume of Sample stock solution with Buffer No. 10 to obtain a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed for amphotericin B under AntibioticsMicrobial Assays 81.
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Official Monographs / Ampicillin 233

Pre-column: 4-mm 5-cm; 5- to 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for ampicillin and caffeine are 0.5 and 1.0, respectively, System suitability solution.] Suitability requirements Resolution: NLT 2.0 between the caffeine and ampicillin peaks, System suitability solution. Tailing factor: NMT 1.4 , Standard solution Capacity factor, k: NMT 2.5, Standard solution Relative standard deviation: NMT 2.0% from replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C13H19NO4S in each mg of Ampicillin taken: Result = (rU/rS (CS P/W) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Ampicillin RS in the Standard solution (mg/mL) W = weight, in mg, of Ampicillin taken for the Sample solution (mg/mL) P = stated content of USP Ampicillin RS (g/ml) Acceptance criteria: NLT 900 g and NMT 1050 g SPECIFIC TESTS STERILITY TESTS 71: Where the label states that Ampicillin is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration, except to dissolve 6 g in 800 mL of Fluid D containing sufficient sterile penicillinase to inactivate the ampicillin and to swirl the vessel until solution is complete before filtering. CRYSTALLINITY 695: Meets the requirements PH 791: 3.56.0 Sample solution: 10 mg/mL DIMETHYLANILINE 223: Meets the requirements WATER DETERMINATION, Method I 921: NMT 2.0% where it is labeled as Ampicillin (anhydrous); between 12.0% and 15.0% where it is labeled as Ampicillin (trihydrate) BACTERIAL ENDOTOXINS TEST 85: Where the label states that Ampicillin is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.15 USP Endotoxin Unit/mg of ampicillin. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label to indicate whether it is anhydrous or is the trihydrate. Where the quantity of ampicillin is indicated in the labeling of any preparation containing Ampicillin, this shall be understood to be in terms of anhydrous ampicillin (C16H19N3O4S). Where it is intended for use in preparing injectable dosage forms, the label states that it is the trihydrate and that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Ampicillin Trihydrate RS USP Endotoxin RS rU rS CS
SPECIFIC TESTS MINIMUM FILL 755: Meets the requirements WATER DETERMINATION, Method I 921: NMT 1.0%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes, or other well-closed containers. USP REFERENCE STANDARDS 11 USP Amphotericin B RS
Ampicillin
(Comment on this Monograph)id=m4430=Ampicillin=AMonos.pdf)
349.41 C16H19N3O4S (anhydrous) 4-Thia-1-azabicyclo[3.2.0]heptane-2 carboxylic acid, [6(aminophenylacetyl)amino]-3,3-dimethyl-7-oxo-, [2S[2,5,6(S*)]]-; (2S,5R,6R)-6-[(R)-2-Amino-2-phenylacetamido]-3,3-dimethyl-7oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid [69-53-4]. Trihydrate 403.46 [7177-48-2]. DEFINITION Ampicillin is anhydrous or contains three molecules of water of hydration. It contains NLT 900 g and NMT 1050 g of C16H19N3O4S per mg, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K: except that where the specimen under test is the trihydrate, both it and the USP Ampicillin Trihydrate RS are undried. ASSAY PROCEDURE Mobile phase: Acetonitrile, water, 1 M monobasic potassium phosphate, and 1 N acetic acid (80:909:10:1) Diluent: Water, 1 M monobasic potassium phosphate, and 1 N acetic acid (989:10:1) Standard solution: 1 mg/mL of USP Ampicillin RS in Diluent. [NOTEDissolve the sample by shaking and sonication, if necessary, to dissolve. Use this solution promptly after preparation.] System suitability solution: 0.12 mg/mL of caffeine in Standard solution Sample solution: Equivalent to 1 mg/mL of anhydrous ampicillin from Diluent. [NOTEAdd a sufficient quantity of Diluent, shake and sonicate, if necessary, to dissolve and then dilute to volume. Use this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm analytical column containing 5to 10-m packing L1
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Ampicillin Boluses
(Comment on this Monograph)id=m4433=Ampicillin Boluses=AMonos.pdf) DEFINITION Ampicillin Boluses contain an amount of ampicillin (as the trihydrate) equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of ampicillin (C16H19N3O4S). IDENTIFICATION PROCEDURE Standard solution: 5 mg/mL of USP Ampicillin RS in acetone and 0.1 N hydrochloric acid (4:1) Sample solution: Equivalent of 10 mg/mL of ampicillin, from 1 or more powdered Boluses, in acetone and 0.1 N hydrochloric acid (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2 L Developing solvent system: Acetone, toluene, glacial acetic acid, and water (26:4:1:4) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, mark the solvent front, and allow to air-dry. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 90 for 15 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Standard solution: Prepare as directed for Standard Preparation under Iodometric AssayAntibiotics 425, using USP Ampicillin RS. Sample solution: Place NLT 5 Boluses in a high-speed glass blender jar containing a volume of water, and blend for 4 1 min. Dilute an accurately measured volume of this stock solution quantitatively and stepwise with water to obtain a Sample solution containing 1.25 mg/mL of ampicillin. Analysis Samples: Standard solution and Sample solution Proceed as directed for Procedure under Iodometric Assay Antibiotics 425. Calculate as A the quantity, in mg, of C16H19N3O4S in each Bolus taken: A = (T/D)(F/2000)(B I) T D = labeled quantity of ampicillin in each Bolus (mg) = concentration of ampicillin in the Sample solution on the basis of the labeled quantity in each Bolus and the extent of dilution (mg/mL) Calculate the quantity, in percentage, of C16H19N3O4S in each Bolus taken: Result = (A/L) 100 A L = as defined above = label claim (mg)
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: SPECIFIC TESTS LOSS ON DRYING 731: NMT 5.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Boluses to indicate that they are for veterinary use only. USP REFERENCE STANDARDS 11 USP Ampicillin RS
Ampicillin Capsules
(Comment on this Monograph)id=m4440=Ampicillin Capsules=A-Monos.pdf) DEFINITION Ampicillin Capsules contain an amount of ampicillin (anhydrous or as the trihydrate) equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of ampicillin (C16H19N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 5 mg/mL of USP Ampicillin RS in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Sample solution: 5 mg/mL of ampicillin, from Powdered Capsules, in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2 L Developing solvent system: Acetone, toluene, glacial acetic acid, and water (26:4:1:4) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution Apply the Sample solution and the Standard solution and develop the chromatogram. When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, mark the solvent front, and allow to air-dry. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 90 for 15 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Standard solution: Prepare as directed for Standard Preparation under Iodometric AssayAntibiotics 425, using USP Ampicillin RS. Sample solution: Place NLT 5 Capsules in a high-speed glass blender jar containing an accurately measured volume of water, and blend for 4 1 min. Dilute an accurately measured volume of this stock solution quantitatively and stepwise with water to obtain a Sample solution containing 1.25 mg/mL of ampicillin.
USP 32
Analysis: Proceed as directed for Procedure under Iodometric AssayAntibiotics 425. Calculate the quantity, in mg, of C16H19N3O4S in each Capsule taken: Result = (T/D) (F/2000) (B I) = labeled quantity, in mg, of ampicillin in each Capsule D = concentration of ampicillin in the Sample solution on the basis of the labeled quantity in each Capsule and the extent of dilution (mg/mL) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Standard solution: L/900 mg/mL of USP Ampicillin RS in water, L being the labeled amount, in mg, of ampicillin/Capsule Analysis: Proceed as directed for Procedure in Automated Methods of Analysis 16, AntibioticsHydroxylamine Assay, using a filtered portion of the solution under test as the Sample solution. Calculate the quantity, in mg, of C16H19N3O4S dissolved: 0.9 C P (AU/AS) Tolerances: NLT 75% (Q) of the labeled amount of C16H19N3O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 4.0% where the Capsules contain anhydrous ampicillin, or between 10.0% and 15.0% where the Capsules contain ampicillin trihydrate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label to indicate whether the ampicillin therein is in the anhydrous form or is the trihydrate. USP REFERENCE STANDARDS 11 USP Ampicillin RS T

Constitute Ampicillin for Injection in a volume of Diluent, corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and quantitatively dilute with Diluent. [NOTEUse this solution promptly after preparation.] Sample solution B (where the label states the quantity of ampicillin in a given volume of constituted solution): 1 mg/mL of ampicillin in Diluent Constitute 1 container of Ampicillin for Injection in a volume of Diluent, corresponding to the volume of solvent specified in the labeling. Quantitatively dilute an accurately measured portion of the constituted solution with Diluent. [NOTEUse this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30 cm analytical column containing 5to 10-m packing L1 Precolumn: 4-mm 5-cm; 5- to 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for ampicillin and caffeine are 0.5 and 1.0, respectively, System suitabilty solution. ] Suitability requirements Resolution: NLT 2.0 between the caffeine and the ampicillin, System suitabilty solution. System suitability solution: 0.12 mg/mL of caffeine in Standard solution Tailing factor: NMT 1.4, Standard solution Capacity factor: NMT 2.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in mg, of ampicillin (C16H19N3O4S) in the container and in the volume of constituted solution taken: Result = (rU/rS) (CS P/1000) (L/D) = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ampicillin RS in the Standard solution (mg/mL) D = concentration, in mg of ampicillin (C16H19N3O4S)/mL, of Sample solution A or Sample solution B, based on the labeled quantity in the container or in the portion of constituted solution taken, respectively, and the extent of dilution L = labeled quantity of ampicillin (C16H19 N3O4S) in the container, or in the volume of constituted solution taken (mg) P = stated content of USP Ampicillin RS (g/mg) [NOTEWhere the test for Uniformity of dosage units has been performed using the Procedure for content uniformity, use the average of these determinations as the Assay value.] Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements. Procedure for content uniformity: Perform the assay on individual containers using Sample solution A or Sample solution B, or both, as appropriate. rU rS CS
Ampicillin for Injection

(Comment on this Monograph)id=m4443=Ampicillin for Injection=A-Monos.pdf) DEFINITION Ampicillin for Injection contains an amount of Ampicillin Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of ampicillin (C16H19N3O4S). ASSAY PROCEDURE Mobile phase: Acetonitrile, water, 1 M monobasic potassium phosphate, and 1 N acetic acid (80:909:10:1) Diluent: Water, 1 M monobasic potassium phosphate, and 1 N acetic acid (989:10:1) Standard solution: 1 mg/mL of USP Ampicillin RS in Diluent [NOTEDissolve the Standard by shaking and sonication, if necessary, to dissolve. Use this solution promptly after preparation.] System suitability solution: 0.12 mg/mL of caffeine in Standard solution Sample solution A (where it is represented as being in a single-dose container): 1 mg/mL of ampicillin in Diluent.
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Analysis: Proceed as directed under Iodometric Assay Antibiotics 425. Calculate the quantity, in mg, of C16H19N3O4S in the portion of Soluble Powder taken: (F/20)(B I) Terms as defined underIodometric AssayAntibiotics 425 Acceptance criteria: 90.0%120.0% SPECIFIC TESTS PH 791: 3.56.0 Sample solution: equivalent of 20 mg/mL of ampicillin WATER DETERMINATION, Method I 921: NMT 5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Ampicillin RS
SPECIFIC TESTS OTHER REQUIREMENTS: It meets the requirements of the test for Identification under Ampicillin Sodium. It also meets the requirements under Injections 1, Labeling. CRYSTALLINITY 695: Meets the requirements. [NOTEAmpicillin Sodium in the freeze-dried form is exempt from this requirement.] PH 791: 8.010.0, in a solution containing 10.0 mg/mL of ampicillin Sample solution: 10.0 mg/mL WATER DETERMINATION, Method I 921: NMT 2.0% PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections. STERILITY TESTS 71: Meets the requirements. BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin Unit/mg of ampicillin CONSTITUTED SOLUTION: At the time of use, it meets the requirements for InjectionsConstituted Solutions 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. Protect the constituted solution from freezing. USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Ampicillin Sodium RS USP Endotoxin RS
Ampicillin for Injectable Suspension

(Comment on this Monograph)id=m4474=Ampicillin for Injectable Suspension=A-Monos.pdf) DEFINITION Ampicillin for Injectable Suspension is a dry mixture of ampicillin trihydrate and one or more suitable buffers, preservatives, stabilizers, and suspending agents. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of ampicillin (C16H19N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 5 mg/mL of USP Ampicillin RS in acetone and 0.1 N hydrochloric acid (4:1) Sample solution: 5 mg/mL of ampicillin in acetone and 0.1 N hydrochloric acid (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2 L Developing solvent system: Acetone, toluene, glacial acetic acid, and water (26:4:1:4) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Sample solution and Standard solution When the solvent front has moved three-fourths of the length of the plate, remove the plate from the chamber, mark the solvent front, and allow to air-dry. Locate the spots on the plate by spraying lightly with Spray reagent, and dry at 90 for 15 min. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Phosphate buffer solution: 136 mg/mL of monobasic potassium phosphate Mobile phase: Acetonitrile, glacial acetic acid, water, and Phosphate buffer solution (90:1:900:10). Pass through a 0.45m nylon filter, and degas. Standard solution: 0.5 mg/mL of USP Ampicillin RS. Dissolve with sonication. Pass through a 0.45-m PTFE filter, discarding the first 3 mL of the filtrate.
Ampicillin Soluble Powder

(Comment on this Monograph)id=m4448=Ampicillin Soluble Powder=A-Monos.pdf) DEFINITION Ampicillin Soluble Powder is a dry mixture of Ampicillin (as the trihydrate) and one or more suitable diluents and stabilizing agents. It contains NLT 90.0% and NMT 120.0% of the labeled amount of ampicillin (C16H19N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 5 mg/mL of USP Ampicillin RS in acetone and 0.1 N hydrochloric acid (4:1) Sample solution: 10 mg/mL of ampicillin from Soluble Powder in acetone and 0.1 N hydrochloric acid (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2 L Developing solvent system: Acetone, toluene, glacial acetic acid, and water (26:4:1:4) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Sample solution and Standard solution When the solvent front has moved three-fourths of the length of the plate, remove the plate from the chamber, mark the solvent front, and allow to air-dry. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 90 for 15 min. Acceptance criteria: The RF value of the principal spot obtained from the solution under test corresponds to that obtained from the Standard solution. ASSAY PROCEDURE Standard solution: Prepare as directed under Iodometric AssayAntibiotics 425, using USP Ampicillin RS. Sample solution: Transfer a quantity of Soluble Powder, equivalent to 125 mg of ampicillin, to a 100-mL volumetric flask, dissolve in and dilute with water to volume.
USP 32
Caffeine solution: 30 mg of caffeine in a 50-mL volumetric flask. Add 25 mL of water, sonicate to dissolve, and dilute with water to volume. Pass through a 0.45-m PTFE filter, discarding the first 3 mL of the filtrate. System suitability solution: Caffeine solution and Standard solution (1:9) Sample solution: Dilute a volume of Ampicillin for Injectable Suspension, constituted as directed in the labeling, with water to obtain a solution containing 0.5 mg/mL of ampicillin. Pass through a 0.45-m PTFE filter, discarding the first 3 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 10-m packing L1 Temperature: 40 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Order of elution: Ampicillin followed by caffeine Resolution: Greater than 2 between ampicillin and caffeine, System suitability solution Column efficiency: NLT 2000 theoretical plates for the ampicillin peak, Standard solution Tailing factor: NMT 1.4, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of C16H19N3O4S in each mL of the constituted solution of Ampicillin for Injectable Suspension taken: Result = (rU/rS) (CS/CU) 100 = average peak response of ampicillin from the Sample solution = average peak response of ampicillin from the rS Standard solution = concentration of USP Ampicillin RS in the CS Standard solution (mg/mL) = nominal concentration of ampicillin in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% rU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Endotoxin RS
Ampicillin for Oral Suspension

(Comment on this Monograph)id=m4480=Ampicillin for Oral Suspension=A-Monos.pdf) DEFINITION Ampicillin for Oral Suspension contains an amount of Ampicillin (anhydrous or as the trihydrate) equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of C16H19N3O4S, when constituted as directed. It contains one or more suitable buffers, colors, flavors, preservatives, and sweetening ingredients. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 5 mg/mL of USP Ampicillin RS in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Sample solution: 5 mg/mL of ampicillin in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2 L Developing solvent system: Acetone, toluene, glacial acetic acid, and water (26:4:1:4) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution Apply the Standard solution and the Sample solution and develop the chromatogram. When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, mark the solvent front, and allow to air-dry. Locate the spots on the plate by spraying lightly with Spray reagent and dry at 90 for 15 min: the RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Standard solution: Prepare as directed for Standard solution under Iodometric AssayAntibiotics 425, using USP Ampicillin RS. Sample solution: Dilute an accurately measured volume of Ampicillin for Oral Suspension, constituted as directed in the labeling, freshly mixed and free from air bubbles, quantitatively and stepwise with water to obtain a solution containing 1.25 mg/mL of ampicillin. Analysis: Proceed as directed for Iodometric Assay Antibiotics 425, Procedure. Calculate the quantity, in mg, of C16H19N3O4S in each mL of the constituted suspension prepared from Ampicillin for Oral Suspension taken: Result = (T/D) (F/2000) (BI) T D = the labeled quantity of ampicillin in the constiuted suspension (mg/ml) = concentration of ampicillin in the Sample solution on the basis of the labeled quantity in the constiuted suspension and the extent of dilution (mg/mL)
SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Antibiotic Solids, Bulks, and Blends in the section Test for Sterility of the Product to Be Examined, Membrane Filtration, except to use Fluid D, to which has been added sufficient sterile penicillinase to inactivate the ampicillin and to swirl the vessel until solution is complete before filtering. If it does not dissolve completely, proceed as directed for Solids in the section Test for Sterility of the Product to be Examined, Direct Inoculation of the Culture Medium, except to use Fluid Thioglycollate Medium and SoybeanCasein Digest Medium containing sufficient penicillinase to inactivate the ampicillin in each vessel. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.15 Endotoxin Unit/mg of ampicillin. PH 791: 5.07.0, in the suspension constituted as directed in the labeling WATER DETERMINATION Method I 921: 11.4%14.0% OTHER REQUIREMENTS: Meets the requirements for Labeling under Injections 1.
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90.0%120.0%
USP 32
Analysis Samples: Standard solution and Sample solution Proceed as directed under Procedure. Calculate the quantity, in mg, of C16H19N3O4S in each Tablet taken: Result = (T/D) (F/2000) (BI) = labeled quantity of ampicillin in each Tablet (mg) = concentration of ampicillin in the Assay Preparation on the basis of the labeled quantity in each Tablet and the extent of dilution (mg/mL) F = conversion factor (g) B = volume of 0.01 N sodium thiosulfate consumed in the Blank Determination (g) I = volume of 0.01 N sodium thiosulfate consumed in the Inactivation and Titration (mL) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Standard solution: L/900 mg/mL of USP Ampicillin RS in water, L being the labeled amount, in mg, of ampicillin per Tablet Analysis Samples: Standard solution and Sample solution Proceed as directed under Procedure in the section Automated Methods of Analysis 16, Antibiotics Hydroxylamine Assay, using a filtered portion of the solution under test as the Sample solution. Calculate the quantity, in mg, of C16H19N3O4S dissolved: 0.9 C P (AU/AS) = concentration of USP Ampicillin RS in the Standard solution (mg/mL) P = stated content of USP Ampicillin RS (g/mg) = absorbance of the Sample solution AU = absorbance of the Standard solution AS Tolerances: NLT 75% (Q) of the labeled amount of C16H19N3O4S is dissolved UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 4.0% where nonchewable Tablets contain anhydrous Ampicillin; between 9.5% and 12.0% where nonchewable Tablets contain Ampicillin trihydrate; NMT 3.0% where chewable Tablets contain anhydrous Ampicillin; NMT 5.0% where chewable Tablets contain Ampicillin trihydrate; and NMT 13.0% where Tablets are labeled for veterinary use only and contain Ampicillin trihydrate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label the Tablets to indicate whether the ampicillin therein is in the anhydrous form or is the trihydrate. Label chewable Tablets to indicate that they are to be chewed before swallowing. Tablets intended for veterinary use only are so labeled. USP REFERENCE STANDARDS 11 USP Ampicillin RS C T D
PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905 For solid packaged in single-unit containers: Meets the requirements SPECIFIC TESTS PH 791: 5.07.5, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 2.5%, or NMT 5.0% if it contains ampicillin trihydrate and contains the equivalent of 100 mg/mL of ampicillin when constituted as directed in the labeling ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label to indicate whether the ampicillin therein is in the anhydrous form or is the trihydrate. USP REFERENCE STANDARDS 11 USP Ampicillin RS
Ampicillin Tablets
(Comment on this Monograph)id=m4490=Ampicillin Tablets=AMonos.pdf) DEFINITION Ampicillin Tablets contain an amount of Ampicillin (anhydrous form or trihydrate form) equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of ampicillin (C16H19N3O4S). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 5 mg/mL of USP Ampicillin RS in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Sample solution: 5 mg/mL of ampicillin, from Powdered Tablets, in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2 L Developing solvent system: Acetone, toluene, glacial acetic acid, and water (26:4:1:4) Spray reagent: 3 mg/mL of ninhydrin in alcohol Analysis Samples: Standard solution and Sample solution Apply the Standard solution and the Sample solution and develop the chromatogram using the Developing solvent system, until the solvent front has moved about threefourths of the length of the plate. Locate the spots on the plate by spraying lightly with the Spray reagent and dry at 90 for 15 min. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY IODOMETRIC ASSAYANTIBIOTICS 425 Standard solution: Prepare as directed under Standard preparation, using USP Ampicillin RS. Sample solution: Place NLT 5 Tablets in a high-speed glass blender jar containing an accurately measured volume of water, and blend for 4 1 min. Dilute an accurately measured volume of this stock solution with water to obtain a concentration of 1.25 mg/mL of ampicillin.
USP 32

= absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Probenecid RS in the Standard solution (mg/mL) = nominal concentration of probenecid in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for Solids in Single-Unit Containers under Content Uniformity with respect to ampicillin and probenecid. DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 5.07.5, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, unit-dose containers. USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Probenecid RS AU AS CS
Ampicillin and Probenecid for Oral Suspension

(Comment on this Monograph)id=m4499=Ampicillin and Probenecid for Oral Suspension=A-Monos.pdf) DEFINITION Ampicillin and Probenecid for Oral Suspension contains an amount of Ampicillin (as the trihydrate) equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of ampicillin (C16H19N3O4S) and NLT 90.0% and NMT 110.0% of the labeled amount of probenecid (C13H19NO4S). It contains one or more suitable colors, flavors, and suspending agents. ASSAY AMPICILLIN Standard solution: Prepare as directed for Standard Preparation under Iodometric AssayAntibiotics 425, using USP Ampicillin RS. Sample stock solution: Constitute Ampicillin and Probenecid for Oral Suspension as directed in the labeling, and mix. Transfer the resulting suspension to a high-speed glass blender jar containing sufficient water to make 500.0 mL, and blend for 10 min. Sample solution: Quantitatively dilute a measured volume of the Sample stock solution with water to obtain a Sample solution containing nominally 1.25 mg/mL of ampicillin. Analysis Proceed as directed for Procedure under Iodometric AssayAntibiotics 425. Calculate the quantity, in mg, of C16H19N3O4S in the portion of Ampicillin and Probenecid for Oral Suspension taken: Result = (F/CU) (VB VU) (1/VS) (100/L) = equivalent of ampicillin/mL of titrant (g/mL) = nominal concentration of ampicillin in the Sample solution (mg/mL) = volume of Blank titer (mL) VB = volume of Sample titer (mL) VU = volume of Sample solution used in the titration VS L = label claim of ampicillin in the Ampicillin and Probenecid for Oral Suspension (g/mg) Acceptance criteria: 90.0%120.0% PROBENECID Standard solution: 1 mg/mL of USP Probenecid RS in sodium carbonate solution (1 in 100) Sample solution: Constitute Ampicillin and Probenecid for Oral Suspension as directed in the labeling, and mix. Quantitatively dilute the resulting suspension with the sodium carbonate solution (1 in 100) to obtain a solution containing nominally 1 mg/mL of probenecid. Transfer 2.0 mL of the clear Sample solution to a 125-mL separator, and add 8.0 mL of 1.0 N hydrochloric acid. Extract this solution with four 20-mL portions of chloroform, filtering each extract through a glass wool pledget and 6 g of chloroform-washed anhydrous sodium sulfate into a 100-mL volumetric flask. Wash the pledget and the sodium sulfate with chloroform, collecting the washings in the 100-mL volumetric flask, dilute with chloroform to volume, and mix. Treat 2.0 mL of the Standard solution in the same manner. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 257 nm Blank: Chloroform washed with sodium carbonate solution (1 in 100) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C13H19NO4S in the Ampicillin and Probenecid for Oral Suspension taken: Result = (AU/AS) (CS/CU) 100 F CU
Ampicillin Sodium
(Comment on this Monograph)id=m4510=Ampicillin Sodium=A-Monos.pdf) C16H18N3NaO4S 371.39 4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, [6(aminophenylacetyl)amino]-3,3-dimethyl-7-oxo-, monosodium salt, [2S-[2,5,6(S*)]]-; Monosodium D-(-)-6-(2-amino-2-phenylacetamido)-3,3dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2carboxylate [69-52-3]. DEFINITION Ampicillin Sodium has a potency equivalent to NLT 845 g and NMT 988 g of (C16H19N3O4S) per mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. IDENTIFICATION TESTSGENERAL, Sodium 191 ASSAY PROCEDURE Diluent: Water, 1 M monobasic potassium phosphate, and 1 N acetic acid (989:10:1) Mobile phase: Acetonitrile, water, 1 M monobasic potassium phosphate, and 1 N acetic acid (80:909:10:1) Standard solution: 1 mg/mL of USP Ampicillin RS in Diluent using shaking and sonication, if necessary, to dissolve. Use this solution promptly after preparation. System suitability solution: 0.12 mg/mL of caffeine in Standard solution Sample solution: [NOTEAmpicillin Sodium is hygroscopic. Minimize exposure to the atmosphere, and weigh promptly.] Equivalent to 1 mg/mL of anhydrous ampicillin in Diluent [NOTEUse this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.)
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RU
USP 32
= ratio of the response of the methylene chloride peak to that of the dioxane peak of the Sample solution = ratio of the response of the methylene chloride RS peak to that of the dioxane peak of the Standard solution = concentration (mg/mL), of methylene chloride CS in the Standard solution = concentration for the Sample solution (mg/mL) CU Acceptance criteria: The limit is 0.2%. DIMETHYLANILINE 223: Meets the requirement, the Internal standard solution, Standard solution, and Sample solution being prepared as follows. Internal standard solution: Dissolve 75 mg of N,Ndiethylaniline in 25 mL of 1 N hydrochloric acid, and dilute quantitatively and stepwise with water to obtain a solution containing 30 g/mL. Standard solution: Transfer 50.0 mg of N,N-dimethylaniline to a 50-mL volumetric flask, add 25 mL of 1 N hydrochloric acid, swirl to dissolve. Dilute with water to volume. Transfer 2.0 mL of the resulting solution to a 100-mL volumetric flask. Dilute with water to volume. To a suitable centrifuge tube add 1.0 mL of this solution, 1.0 mL of 1.25 N sodium hydroxide, 1.0 mL of Internal standard solution, and 1.0 mL of cyclohexane, shake vigorously for 1 min, and centrifuge. Use the clear supernatant. Sample solution: Transfer 1.0 g of Ampicillin Sodium to a centrifuge tube, add 2 mL of 1.25 N sodium hydroxide, swirl to dissolve the sample, add 1.0 mL of Internal standard solution and 1.0 mL of cyclohexane, shake vigorously for 1 min, and centrifuge. Use the clear supernatant. SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements. [NOTEAmpicillin Sodium in the freeze-dried form is exempt from this requirement.] PH 791: 8.010.0 Sample solution: 10.0 mg/mL WATER DETEMINATION, Method I 921: NMT 2.0% STERILITY TESTS 71: Where the label states that Ampicillin Sodium is sterile meets the requirements. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Ampicillin Sodium is sterile or the label states that Ampicillin Sodium must be subjected to further processing during the processing of injectable dosage forms contains NMT 0.15 USP Endotoxin Unit/mg of ampicillin. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Ampicillin Sodium RS USP Endotoxin RS
Mode: LC Detector: UV 254 nm Column Pre-column: 4-mm 5-cm; 5- to 10-m packing L1 Analytical column: 5- to 10-m; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for ampicillin and caffeine are 0.5 and 1.0, respectively, System suitability solution.] Suitability requirements Resolution: NLT 2.0, between the caffeine and the ampicillin peaks, System suitability solution Tailing factor: NMT 1.4, Standard solution Capacity factor: NMT 2.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, of C16H19N3O4S (g) in each mg of Ampicillin Sodium taken: Result = (rU/rS) (CS/CU) P = peak response of the Sample solution = peak response of the Standard solution = concentration, (mg/mL) of USP Ampicillin RS in the Standard solution = concentration for the Sample solution CU (mg/mL) P = stated content of USP Ampicillin RS (g/mg) Acceptance criteria: 845988 g per mg rU rS CS IMPURITIES Organic Impurities PROCEDURE: LIMIT OF METHYLENE CHLORIDE Internal standard solution: 2.1 mg/mL of dioxane in dimethyl sulfoxide Standard solution: 0.33 mg/mL of methylene chloride in Internal standard solution Sample solution: 166.7 mg/mL of Ampicillin Sodium in Internal standard solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.8-m 4-mm glass column packed with a 10% phase G39 on unsilanized support S1A Temperature Column: 65 Injection port: 100 Detector block: 260 Carrier gas: Nitrogen Flow rate: 60 mL/min Injection size: 1 L System suitability Sample: Standard solution [NOTEThe relative retention times for methylene chloride and dioxane are 0.5 and 1.0, respectively] Suitability requirements Resolution: NLT 4 Between the methylene chloride peak and the dioxane peak Relative standard deviation: NMT 5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of methylene chloride in the portion of Ampicillin Sodium taken: Result = (RU/RS) (CS/CU) 100
Ampicillin and Sulbactam for Injection

(Comment on this Monograph)id=m4528=Ampicillin and Sulbactam for Injection=A-Monos.pdf) DEFINITION Ampicillin and Sulbactam for Injection is a sterile, dry mixture of Ampicillin Sodium and Sulbactam Sodium. It contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amounts of ampicillin (C16H19N3O4S) and sulbactam (C8H11NO5S), the labeled amounts representing proportions of ampicillin to sulbactam of 2:1. It contains NLT 563 g of
USP 32
ampicillin and 280 g of sulbactam per mg, calculated on the anhydrous basis. IDENTIFICATION The retention time of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE 0.005 M Tetrabutylammonium hydroxide: Dilute 6.6 mL of a 40% solution of tetrabutylammonium hydroxide with water to obtain 1800 mL of solution. Adjust with 1 M phosphoric acid to a pH of 5.0 0.1, and dilute with water to 2000 mL. Mobile phase: Acetonitrile and 0.005 M Tetrabutylammonium hydroxide (7:33) Standard solution: 0.6 mg/mL of ampicillin and 0.3 mg/mL of sulbactam, obtained from USP Ampicillin RS and USP Sulbactam RS in Mobile phase. [NOTEInject this solution promptly.] System suitability stock solution: Prepare a 0.3 mg/mL solution of USP Sulbactam RS in 0.01 N sodium hydroxide, and allow to stand for 30 min. Adjust with phosphoric acid to a pH of 5.0 0.1. Transfer 5 mL of the solution to a 25mL volumetric flask, add 4.25 mL of acetonitrile, and dilute with 0.005 M Tetrabutylammonium hydroxide to volume. System suitability solution: Transfer 1 mL of System suitability stock solution to a 25-mL volumetric flask, add 15 mg of USP Ampicillin RS, and dilute with Mobile phase to volume. [NOTEInject this solution promptly.] Sample solution A: Mix the contents of a container of Ampicillin and Sulbactam for Injection. Quantitatively dissolve a portion of the powder in the Mobile phase to obtain a solution having a concentration of 1 mg of the powder per mL. [NOTEInject this solution promptly.] Sample solution B (where it is represented as being in a single-dose container): Constitute a container of Ampicillin and Sulbactam for Injection with a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw the total withdrawable contents from the container, using a suitable hypodermic needle and syringe, and dilute quantitatively, and stepwise if necessary, with Mobile phase to obtain a solution containing nominally 0.6 mg/mL of ampicillin and 0.3 mg/mL of sulbactam. [NOTE Inject this solution promptly.] Sample solution C (where the label states the quantities of ampicillin and sulbactam in a given volume of constituted solution): Constitute a container of Ampicillin and Sulbactam for Injection with a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a volume of the constituted solution quantitatively, and stepwise if necessary, with the Mobile phase to obtain a solution containing 0.6 mg/mL of ampicillin and nominally 0.3 mg/mL of sulbactam. [NOTE Inject this solution promptly.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for ampicillin for sulbactam alkaline degradation product are 0.7 and 1.0, respectively, System suitability solution; for ampicillin and sulbactam are 0.35 and 1.0, respectively, Standard solution.]

Suitability requirements Resolution: NLT 4.0 between ampicillin and sulbactam alkaline degradation product, System suitability solution Column efficiency: NLT 3500 theoretical plates, from the sulbactam peak, Standard solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and the appropriate Sample solution Calculate the percentage of C16H19N3O4S and of C8H11NO5S in the portion of Ampicillin and Sulbactam for Injection taken: Result = (rU/rS) (CS/CU) P F 100 = peak area for the appropriate analyte from Sample solution A rS = peak area from the Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) = concentration of Sample solution A (mg/mL) CU P = stated content of the appropriate USP Reference Standard (g/mg) F = unit conversion factor; 0.001 mg/g Calculate the quantities of C16H19N3O4S and of C8H11NO5S withdrawn from the container, or in the volume of constituted solution taken: Result = (rU/rS) (CS/CU) P F 100 = peak area for the appropriate analyte from Sample solution B or Sample solution C = peak area from the Standard solution rS CS = concentration of the appropriate USP Reference Standard in the Standard solution (mg/mL) = nominal concentration of ampicillin or sulbactam CU (mg/mL) P = stated content of the appropriate USP Reference Standard (g/mg) F = unit conversion factor; 0.001 mg/g Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements rU rU
SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791: 8.010.0, in a solution containing 10 mg of ampicillin and 5 mg of sulbactam per mL. Sample solution: 10 mg/mL ampicillin and 5 mg/ml sulbactam WATER DETERMINATION, Method I 921: NMT 2.0% PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.17 USP Endotoxin Unit in a portion equivalent to 1 mg of a mixture of ampicillin and sulbactam (0.67 and 0.33 mg, respectively). CONSTITUTED SOLUTION: At the time of use, it meets the requirements for InjectionsConstituted Solutions 1. OTHER REQUIREMENTS: It meets the requirements for InjectionsLabeling 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in InjectionsContainers for Sterile Solids 1.
242

rU
USP 32
= amprolium peak response from the Sample solution = amprolium peak response from the Standard rS solution = concentration of USP Amprolium RS in the CS Standard solution (mg/mL) CU = concentration of Amprolium in the Sample solution (mg/mL) Acceptance criteria: 97.0%101.0% SPECIFIC TESTS LOSS ON DRYING 731: Dry it at a pressure not exceeding 5 mm of mercury at 100 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Amprolium RS
USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Endotoxin RS USP Sulbactam RS
Amprolium
(Comment on this Monograph)id=m4540=Amprolium=AMonos.pdf)
315.24 C14H19ClN4 HCl 1-[(4-Amino-2-propyl-5-pyrimidinyl)methyl]-2-methylpyridinium chloride monohydrochloride; 1-[(4-Amino-2-propyl-5-pyrimidinyl)methyl]-2-picolinium chloride monohydrochloride [121-25-5]. DEFINITION Amprolium contains NLT 97.0% and NMT 101.0% of amprolium (C14H19ClN4 HCl), calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K: Previously dried B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 246 nm Sample solution: 10 g/mL in 0.1 N hydrochloric acid Acceptance criteria: Absorptivities calculated on the dried basis do not differ by more than 3.0%. ASSAY PROCEDURE Diluent: Methanol, acetonitrile, and water (9:1:10) Mobile phase: 6 g of sodium 1-heptanesulfonate in 500 mL of water, add 12 mL of glacial acetic acid, 2.0 mL of triethylamine, 450 mL of methanol, and 50 mL of acetonitrile. Pass through a suitable filter of 0.5-m or finer porosity. System suitability solution: 0.5 mg/mL of USP Amprolium RS and 0.2 mg/mL of 2-picoline in Diluent Standard solution: 0.5 mg/mL of USP Amprolium RS in Diluent Sample solution: 0.5 mg/mL of Amprolium in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L13 Flow rate: 0.6 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 7 between amprolium and 2-picoline Column efficiency: NLT 6500 theoretical plates from the amprolium peak Tailing factor: NMT 2.3 for the analyte peak Relative standard deviation: NMT 1.0% for amprolium Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H19ClN4 HCl in the portion of Amprolium taken: Result = (rU/rS) (CS/CU) 100
Amprolium Soluble Powder

(Comment on this Monograph)id=m4547=Amprolium Soluble Powder=A-Monos.pdf) DEFINITION Amprolium Soluble Powder contains NLT 95.0% and NMT 105.0% of the labeled amount of amprolium (C14H19ClN4 HCl). IDENTIFICATION ULTRAVIOLET ABSORPTION 197U Sample solution: 10 g/mL (filtered) in 0.1 N hydrochloric acid ASSAY PROCEDURE Diluent: Methanol, acetonitrile, and water (9:1:10) Mobile phase: 6 g of sodium 1-heptanesulfonate in 500 mL of water, add 12 mL of glacial acetic acid, 2.0 mL of triethylamine, 450 mL of methanol, and 50 mL of acetonitrile Pass through a suitable filter of 0.5-m or finer porosity. System suitability solution: 0.5 mg/mL of USP Amprolium RS and 0.2 mg/mL of 2-picoline in Diluent Standard solution: 0.5 mg/mL of USP Amprolium RS in Diluent Sample solution: Nominally equivalent to 0.5 mg/mL of amprolium from Soluble Powder in Diluent [NOTEFirst add a sufficient quantity of Diluent to Soluble Powder and sonicate for 10 min. Allow to cool to room temperature, dilute with Diluent to volume. Pass through a suitable filter of 0.5-m or finer porosity, and use the clear filtrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L13 Flow rate: 0.6 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 7 between amprolium and 2-picoline Column efficiency: NLT 6500 theoretical plates from the amprolium peak
USP 32
Tailing factor: NMT 2.3 for the analyte peak Relative standard deviation: NMT 1.0% amprolium Analysis Samples: Sample solution and Standard solution Calculate the percentage of C14H19ClN4 HCl in the portion of Soluble Powder taken: Result = (rU/rS) (CS/CU) 100 = amprolium peak response obtained from the Sample solution = amprolium peak response obtained from the rS Standard solution CS = concentration of USP Amprolium RS in the Standard solution (mg/mL) CU = nominal concentration of amprolium in the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Amprolium RS rU rS
Official Monographs / Amyl 243

= amprolium peak response from the Standard solution = concentration of USP Amprolium RS in the CS Standard solution (mg/mL) CU = nominal concentration of amprolium oral solution in the Sample solution (mg/mL) Acceptance criteria: 93.0%107.0% SPECIFIC TESTS PH 791: 2.53.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. Store at a temperature between 5 and 30, in a dry place. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Amprolium RS
Amyl Nitrite
(Comment on this Monograph)id=m4570=Amyl Nitrite=AMonos.pdf) C5H11NO2 117.15 Mixture of nitrous acid, 2-methylbutyl ester, and nitrous acid, 3-methylbutyl ester [8017-89-8; 110-46-3]. DEFINITION Amyl Nitrite is a mixture of the nitrite esters of 3-methyl-1butanol and 2-methyl-1-butanol. It contains NLT 85.0% and NMT 103.0% of C5H11NO2. [CAUTIONAmyl Nitrite is very flammable. Do not use where it may be ignited.] IDENTIFICATION A. The NMR spectrum recorded as directed in the Assay exhibits, among other peaks, a doublet with a band centered at about 1 ppm and a multiplet with a band centered at about 4.8 ppm representing methyl protons and methylene protons alpha to the nitrite group, respectively, both relative to the tetramethylsilane singlet at 0 ppm. B. To a few drops of it, add a mixture of 1 mL of ferrous sulfate TS and 5 mL of 3 N hydrochloric acid. Acceptance criteria: A greenish brown color is produced. ASSAY PROCEDURE Diluent: Carbon tetrachloride Internal standard: USP Benzyl Benzoate RS Analysis: Transfer 4 to 5 mEq of Internal standard to a semimicro sampling tube, add 2 to 3 mL of Diluent, apply a sampling valve and septum,* thereby sealing the tube, and determine the weight of the sealed assembly. Open the valve, introduce about 500 L of Amyl Nitrite with a syringe, close the valve, and determine the weight of the sealed assembly when it has attained constant weight. Shake the sampling tube and valve assembly, and transfer about 500 L of the solution to a precision NMR tube as directed for Nuclear Magnetic Resonance 761, Absolute Method of Quantitation. With no spinning, or with the spinning adjusted so that the spinning side bands of neither the substance under assay nor the Internal standard interfere with the regions to be integrated, record as AS the average area of the Internal standard singlet appearing at about 5.3 ppm, representing the methylene protons of benzyl *Suitable sampling tubes, sampling valves, and septums are available,
respectively, as catalog Nos. K-749000, K-749100, and K-749102 (50 septums) or K-749101 (100 septums), from Kontes Glass Company, Vineland, NJ 08360.
Amprolium Oral Solution

(Comment on this Monograph)id=m4548=Amprolium Oral Solution=A-Monos.pdf) DEFINITION Amprolium Oral Solution contains NLT 93.0% and NMT 107.0% of the labeled amount of amprolium (C14H19ClN4 HCl). IDENTIFICATION ULTRAVIOLET ABSORPTION 197U Solution: 10 g/mL (filtered), in 0.1 N hydrochloric acid ASSAY PROCEDURE Mobile phase: To 4.5 g of sodium 1-hexanesulfonate add 1500 mL of water, 400 mL of methanol, and 100 mL of acetonitrile, mix, and allow to cool to room temperature. Adjust with phosphoric acid to a pH of 5.1, and pass through a filter having a 0.5-m or finer porosity. Standard solution: 0.5 mg/mL of USP Amprolium RS Sample solution: Equivalent to 0.48 mg/mL of amprolium, from Oral Solution diluted with water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 268 nm Column: 3.9-mm 30-cm; packing L11 Temperature: 45 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H19ClN4 HCl in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 rU = amprolium peak response from the Sample solution
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USP 32
ASSAY PROCEDURE Diluent: Carbon tetrachloride Internal standard: USP Benzyl Benzoate RS Analysis: Remove the gauze or other covering from one or more Inhalant ampuls containing a total of 300 to 400 L of amyl nitrite. Weigh accurately the clean and dry intact glass ampul(s), and place the weighed specimen in a freezer for NLT 15 min. Transfer the chilled specimen to a glassstoppered, 25-mL conical flask containing a solution of 4 to 5 mEq of Internal standard in 1 to 2 mL of Diluent. Break the ampul(s) with a glass rod, and rinse any sample or glass fragments adhering to the glass rod with 1 mL of Diluent into the main Assay solution. Insert the stopper in the flask immediately and proceed as directed for Nuclear Magnetic Resonance 761, Absolute Method of Quantitation, beginning with When dissolution has been completed. With no spinning, or with the spinning adjusted so that the spinning side bands of neither the substance under Assay nor the Internal standard interfere with the regions to be integrated, record as AS the average area of the Internal standard singlet appearing at about 5.3 ppm, representing the methylene protons of benzyl benzoate, and record as AU the average area of the multiplet with a band center at about 4.8 ppm, representing the alpha methylene protons of amyl nitrite, with reference to the tetramethylsilane singlet at 0 ppm. Calculate the quantity of C5H11NO2 in the Inhalant taken, using 58.57 as the equivalent weight of amyl nitrite (EWU) and 106.12 as that of benzyl benzoate (EWS). Rinse the flask containing the assay preparation with three 5-mL portions of ether, decanting each rinsing carefully to avoid loss of glass fragments, and evaporate any remaining ether with the aid of a current of dry air. Transfer the dry glass fragments to a tared watch glass, weigh, and subtract the weight of the glass fragments from that of the intact ampul(s) to obtain the weight of the Inhalant taken. Acceptance criteria: 80.0%105.0% OTHER COMPONENTS CONTENT OF TOTAL NITRITES Sample: Remove the gauze or other covering, place the glass container of Inhalant upright in a dry iceacetone slurry, and cool for 10 min. Dry the container of Inhalant, place it in a pointed glass tube, and break the container with a glass rod. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Thermal conductivity Column: Under typical conditions, the instrument contains a 3-mm 2-m column packed with a methyl polysiloxane oil, 25% by weight on suitable calcined diatomite. Column temperature: 80 Injection port and detector block temperature: Maintain at about 10 above the temperature of the column. Carrier gas: Helium Flow rate: 60 mL/min Injection size: NMT 2 L Analysis: From the area under the curve, calculate the percentage (a/a) of total nitrites, represented by the area under the main peak of the chromatogram, in the Amyl Nitrite taken. Acceptance criteria: NLT 95.0% SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.8700.880 ACIDITY Analysis: To 0.30 mL in a glass-stoppered cylinder, add a mixture of 0.60 mL of 0.1 N sodium hydroxide, 10 mL of
benzoate, and record as AU the average area of the multiplet with a band center at about 4.8 ppm, representing the alpha methylene protons of amyl nitrite, with reference to the tetramethylsilane singlet at 0 ppm. Calculate the quantity of C5H11NO2 in the Amyl Nitrite taken, using 58.57 as the equivalent weight of amyl nitrite (EWU) and 106.12 as that of benzyl benzoate (EWS). Acceptance criteria: 85.0%103.0% OTHER COMPONENTS CONTENT OF TOTAL NITRITES Sample: A portion of Amyl Nitrite Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Thermal conductivity Column: Under typical conditions, the instrument contains a 3-mm 2-m column packed with a methyl polysiloxane oil, 25% by weight on suitable calcined diatomite. Column temperature: 80 Injection port and detector block temperature: Maintained at about 10 above the temperature of the column Carrier gas: Helium Flow rate: 60 mL/min Injection size: NMT 2 L Analysis: From the area under the curve, calculate the percentage (a/a) of total nitrites, represented by the area under the main peak of the chromatogram, in the Amyl Nitrite taken. Acceptance criteria: NLT 97.0% IMPURITIES Inorganic Impurities LIMIT OF NONVOLATILE RESIDUE Sample: 10 mL Analysis: Evaporate at room temperature in a tared evaporating dish, in a well-ventilated hood, and dry the residue at 105 for 1 h. Acceptance criteria: The weight of the residue does not exceed 2 mg (0.02%). SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.8700.876 ACIDITY: To 0.30 mL in a glass-stoppered cylinder, add a mixture of 0.60 mL of 0.1 N sodium hydroxide, 10 mL of water, and 1 drop of phenolphthalein TS, and invert the cylinder three times. Acceptance criteria: The red tint of the water layer is still perceptible. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a cool place, protected from light. USP REFERENCE STANDARDS 11 USP Benzyl Benzoate RS
Amyl Nitrite Inhalant

(Comment on this Monograph)id=m4600=Amyl Nitrite Inhalant=A-Monos.pdf) DEFINITION Amyl Nitrite Inhalant contains a mixture of the nitrite esters of 3-methyl-1-butanol and 2-methyl-1-butanol. It contains NLT 80.0% and NMT 105.0% of C5H11NO2. It contains a suitable stabilizer. [CAUTIONAmyl Nitrite Inhalant is very flammable. Do not use where it may be ignited.]
USP 32
water, and 1 drop of phenolphthalein TS, and invert the cylinder three times. Acceptance criteria: The red tint of the water layer is still perceptible. OTHER REQUIREMENTS: It meets the requirements of the Identification tests under Amyl Nitrite. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, unit-dose glass containers, wrapped loosely in gauze or other suitable material, and store in a cool place, protected from light. USP REFERENCE STANDARDS 11 USP Benzyl Benzoate RS
Official Monographs / Anileridine 245

IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221 Sample: 180 mg Analysis: Dissolve the Sample in a mixture of 1 mL of nitric acid and 40 mL of water. Acceptance criteria: NMT 400 ppm; the solution shows no more chloride than corresponds to 0.10 mL of 0.020 N hydrochloric acid. SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1.0%
Anileridine
(Comment on this Monograph)id=m4730=Anileridine=AMonos.pdf)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at room temperature.
Anileridine Injection
(Comment on this Monograph)id=m4760=Anileridine Injection=A-Monos.pdf) 352.47 C22H28N2O2 4-Piperidinecarboxylic acid, 1-[2-(4-aminophenyl)ethyl]-4phenyl-, ethyl ester; Ethyl 1-(p-aminophenethyl)-4-phenylisonipecotate [144-14-9]. DEFINITION Anileridine contains NLT 98.5% and NMT 101.0% of C22H28N2O2, calculated on the anhydrous basis. IDENTIFICATION A. PROCEDURE Sample stock solution: Dissolve 40 mg of Anileridine in 2.3 mL of 0.1 N hydrochloric acid in a 100-mL volumetric flask, and dilute with water to volume. Buffer solution: Dissolve 5.68 g of anhydrous dibasic sodium phosphate and 3.63 g of monobasic potassium phosphate in water to make 1000 mL: the pH is 7.0 0.05. Sample solution A: Sample stock solution, Buffer solution, and water (4:25:71) Sample solution B: Sample stock solution, Buffer solution, and water (20:25:55) Acceptance criteria: The UV absorption spectrum of Sample solution A exhibits a maximum at 234 1 nm; and the UV absorption spectrum of Sample solution B exhibits a maximum at 285 2 nm. The ratio 5A234/A285 is 8.8. B. PROCEDURE Sample solution: 0.2 mg/mL of Anileridine in 0.1 N hydrochloric acid. Analysis: To 5 mL of the Sample solution, add 2 mL of a solution of p-dimethylaminobenzaldehyde in alcohol (1 in 100). Acceptance criteria: A yellow color develops immediately. ASSAY PROCEDURE Sample: 350 mg of Anileridine Analysis: Dissolve the Sample in 50 mL of glacial acetic acid, add 1 drop of crystal violet TS and titrate with 0.1 N perchloric acid VS to a blue-green endpoint. Perform a blank determination (See Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 17.62 mg of C22H28N2O2. DEFINITION Anileridine Injection is a sterile solution of Anileridine in Water for Injection, prepared with the aid of Phosphoric Acid. It contains NLT 90.0% and NMT 115.0% of the labeled amount of anileridine (C22H28N2O2), as the phosphate. IDENTIFICATION A. PROCEDURE Sample solution: 0.25 mg/mL Anileridine from Injection diluted with water Analysis: To 5 mL of the Sample solution, add 2 mL of a solution (1 in 100) of p-dimethylaminobenzaldehyde in alcohol. Acceptance criteria: A yellow color develops immediately. B. A volume of Injection, diluted with water to a concentration of 25 mg of anileridine in 1000 mL, exhibits absorbance maxima at 234 1 and 285 2 nm. ASSAY PROCEDURE Standard solution: 250 g/mL of USP Anileridine Hydrochloride RS in 0.1 N hydrochloric acid Each mg of anileridine hydrochloride is equivalent to 0.8286 mg of anileridine. [NOTEPrepare on the day of the assay.] Sample solution: Nominally equivalent to 200 g/mL of anileridine, from a volume of Injection diluted with 0.1 N hydrochloric acid Blank: 0.1 N hydrochloric acid Spectrometric conditions Analytical wavelength: 560 nm Cell: 1 cm Analysis Samples: Sample solution, Standard solution, and Blank Transfer 5.0 mL each of the Standard solution, the Sample solution, and Blank to separate 200-mL volumetric flasks. To each flask add 25 mL of water, 5 mL of 1 N hydrochloric acid, and 5 mL of sodium nitrite solution (1 in 1000). Allow to stand for 2 min, then add to each flask 5 mL of ammonium sulfamate solution (1 in 200). Allow to stand for 3 min, then add 5 mL of N-(1naphthyl)ethylenediamine dihydrochloride solution (1 in
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1000). Allow to stand for 1 h, and dilute with water to volume. [NOTEUse the reagent blank to set the instrument.] Calculate the percentage of C22H28N2O2 in each mL of the Injection taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100
USP 32
ASSAY PROCEDURE Sample: 200 mg of Anileridine Hydrochloride Analysis: Dissolve the Sample in 10 mL of glacial acetic acid by heating it on a steam bath. Cool immediately in a cold water bath, add 5 mL of mercuric acetate TS, 20 mL of acetone, and 0.5 mL of indicator solution (70 mg of naphtholbenzein, 10 mg of crystal violet, and 40 mg of quinaldine red in 100 mL of glacial acetic acid). Titrate with 0.1 N perchloric acid VS to a gray-green endpoint. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 21.27 mg of C22H28N2O2 2HCl. Acceptance criteria: 96.0%102.0% OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 200 mg Analysis: Dissolve the Sample in 50 mL of water in a glassstoppered flask. Add 25.0 mL of 0.1 N silver nitrate VS, then add 5 mL of 2 N nitric acid and 5 mL of nitrobenzene, shake vigorously, and add 2 mL of ferric ammonium sulfate TS. Titrate the excess silver nitrate with 0.1 N ammonium thiocyanate VS. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Cl. Acceptance criteria: 16.0%17.2% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
= absorbance of the Sample solution = absorbance of the the Standard solution = concentration of USP Anileridine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of anileridine in the CU Sample solution (g/mL) = molecular weight of anileridine, 352.48 Mr1 = molecular weight of anileridine hydrochloride, Mr2 425.40 Acceptance criteria: 90.0%115.0% AU AS CS SPECIFIC TESTS PH 791: 4.55.0 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 7.2 USP Endotoxin Units/mg of anileridine. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Anileridine Hydrochloride RS USP Endotoxin RS
NMT 0.1%
Anileridine Hydrochloride
(Comment on this Monograph)id=m4790=Anileridine Hydrochloride=A-Monos.pdf) C22H28N2O2 2HCl 425.39 4-Piperidinecarboxylic acid, 1-[2-(4-aminophenyl)ethyl]-4phenyl-, ethyl ester, dihydrochloride; Ethyl 1-(p-aminophenethyl)-4-phenylisonipecotate dihydrochloride [126-12-5]. DEFINITION Anileridine Hydrochloride contains NLT 96.0% and NMT 102.0% of C22H28N2O2 2HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample stock solution: 0.5 mg/mL in water pH 7.0 Buffer solution: 5.68 g of anhydrous dibasic sodium phosphate and 3.63 g of monobasic potassium phosphate in water to make 1000 mL: the pH, determined potentiometrically, is 7.0 0.05. Sample solution A: Sample stock solution, pH 7.0 Buffer solution, and water (4:25:71) Sample solution B: Sample stock solution, pH 7.0 Buffer solution, and water (20:25:55) Acceptance criteria: The UV absorption spectrum of Sample solution A exhibits a maximum at 234 1 nm, and the UV absorption spectrum of Sample solution B exhibits a maximum at 285 2 nm. C. PROCEDURE Analysis: To 5 mL of a solution (1 in 5000) add 2 mL of a 1 in 100 solution of p-dimethylaminobenzaldehyde in alcohol. Acceptance criteria: A yellow color develops immediately. D. IDENTIFICATION TESTSGENERAL, Chloride 191: 10 mg/mL
SPECIFIC TESTS PH 791: 2.53.0 Sample solution: 50 mg/mL LOSS ON DRYING 731: Dry it at a pressure below 5 mm of mercury at 100 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Anileridine Hydrochloride RS
Anileridine Hydrochloride Tablets

(Comment on this Monograph)id=m4820=Anileridine Hydrochloride Tablets=A-Monos.pdf) DEFINITION Anileridine Hydrochloride Tablets contain an amount of anileridine hydrochloride (C22H28N2O2 2HCl) equivalent to NLT 95.0% and NMT 105.0% of the labeled amount of anileridine (C22H28N2O2). IDENTIFICATION A. PROCEDURE Sample solution: Transfer an equivalent to 50 mg of anileridine, from finely powdered Tablets, to a 250-mL volumetric flask. Add 100 mL of water, and heat on a steam bath. Cool, dilute to volume, and filter. Analysis: To 5 mL of the Sample solution add 2 mL of a solution (1 in 100) of p-dimethylaminobenzaldehyde in alcohol. Acceptance criteria: A yellow color develops immediately. B. PROCEDURE Sample stock solution: Transfer an equivalent to 50 mg of anileridine, from finely powdered Tablets, to a 100-mL volumetric flask. Add 30 mL of water, and heat on a steam bath. Cool, dilute to volume, and filter.
USP 32
Buffer solution: 5.68 g of anhydrous dibasic sodium phosphate and 3.63 g of monobasic potassium phosphate in water to make 1000 mL: the pH is 7.0 0.05. Sample solution A: Standard stock solution, Buffer solution, and water (4:25:71) Sample solution B: Standard stock solution, Buffer solution, and water (20:25:55) Acceptance criteria: The UV absorption spectrum of Sample solution A exhibits a maximum at 234 1 nm, and the UV absorption spectrum of Sample solution B exhibits a maximum at 285 2 nm. ASSAY PROCEDURE Standard solution: 250 g/mL of USP Anileridine Hydrochloride RS in 0.1 N hydrochloric acid. (Each mg of anileridine hydrochloride is equivalent to 0.8286 mg of anileridine.) [NOTEPrepare on the day of the assay.] Sample solution: Transfer an equivalent to 50 mg of anileridine, from finely powdered Tablets (NLT 20), to a 250mL volumetric flask. Add 25 mL of 1 N hydrochloric acid and 100 mL of water, and heat on a water bath. Cool, and dilute to volume. Filter the solution, discarding the first 25 mL of the filtrate. Blank: 0.1 N hydrochloric acid Spectrometric system Analytical wavelength: 560 nm Cell: 1 cm Analysis Samples: Standard solution, Sample solution, and Blank Transfer 5.0 mL each of the Standard solution, Sample solution, and Blank to separate 200-mL volumetric flasks. To each flask add 25 mL of water, 5 mL of 1 N hydrochloric acid, and 5 mL of sodium nitrite solution (1 in 1000). Allow to stand for 2 min, then add to each flask 5 mL of ammonium sulfamate solution (1 in 200). Allow to stand for 3 min, then add 5 mL of N-(1naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Allow to stand for 1 h, and dilute to volume. [NOTEUse the reagent blank to set the instrument.] Calculate the percentage of C22H28N2O2 in the portion of Tablets taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 = absorbance of the solution from the Sample solution AS = absorbance of the solution from the Standard solution CS = concentration of USP Anileridine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of anileridine in the CU Sample solution (g/mL) = molecular weight of anileridine, 352.48 Mr1 = molecular weight of anileridine hydrochloride, Mr2 425.40 Acceptance criteria: 95.0%105.0% AU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 45 min Standard solution: USP Anileridine Hydrochloride RS in Medium Sample solution: Filtered portion of the solution under test Analysis: Determine the amount of C22H28N2O2 dissolved, using the Analysis set forth in the Assay and in comparison to a Standard solution having a known concentration of USP Anileridine Hydrochloride RS.
Official Monographs / Antazoline 247

Tolerances: NLT 65% (Q) of the labeled amount of C22H28N2O2 is dissolved UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Anileridine Hydrochloride RS
Antazoline Phosphate
(Comment on this Monograph)id=m4920=Antazoline Phosphate=A-Monos.pdf)
363.35 C17H19N3 H3PO4 1H-Imidazole-2-methanamine, 4,5-dihydro-N-phenyl-N(phenylmethyl)-, phosphate (1:1); 2-[(N-Benzylanilino)methyl]-2-imidazoline phosphate (1:1) [154-68-7]. DEFINITION Antazoline Phosphate contains NLT 98.0% and NMT 101.0% of C17H19N3 H3PO4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The RF value of the principal spot of the Identification solution corresponds to that of Standard solution A as obtained in the Procedure under Impurities. ASSAY PROCEDURE Sample solution: 750 mg of Antazoline Phosphate in 50 mL of glacial acetic acid Analysis: Titrate with 0.1 N perchloric acid VS using a glass electrode and a calomel electrode containing a saturated solution of lithium chloride in glacial acetic acid (see Titrimetry 541). Perform a blank determination. Each mL of 0.1 N perchloric acid is equivalent to 36.34 mg of C17H19N3 H3PO4. Acceptance criteria: 98.0%101.0% IMPURITIES Organic Impurities PROCEDURE Standard stock solution: 0.10 mg/mL of USP Antazoline Phosphate RS in methanol Standard solutions: Dilute the Standard stock solution with methanol to obtain five Standard solutions having the following compositions:
Standard Solution A B C D E Concentration (g RS/mL) 50 40 30 20 10 Percentage (for Comparison with Sample) 0.5 0.4 0.3 0.2 0.1
Dilution (1 in 2) (2 in 5) (3 in 10) (1 in 5) (1 in 10)
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USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K: Meets the requirements B. ULTRAVIOLET ABSORPTION 197U Sample solution: 10 g/mL in chloroform ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: n-Hexane, dichloromethane, and glacial acetic acid (41:6:3) Internal standard solution: 500 g/mL of o-nitroaniline in n-hexane [NOTEFirst dissolve o-nitroaniline in a small quantity of dichloromethane, and then dilute with nhexane.] System suitability stock solution: 0.1 mg/mL of USP Anthralin RS and 0.2 mg/mL of danthron in dichloromethane System suitability solution: System suitability stock solution, n-hexane, and Mobile phase (1:1:3) Solvent blank solution: Mobile phase, n-hexane, and dichloromethane (3:1:1) Standard stock solution: 250 g/mL of USP Anthralin RS in dichloromethane Standard solution: Standard stock solution, Internal standard solution, and Mobile phase (1:1:3) Sample stock solution: 250 g/mL of Anthralin in dichloromethane Sample solution: Pipet 10 mL of Sample stock solution into a 100-mL volumetric flask, dilute with dichloromethane to volume, and mix. Pipet 5 mL of this solution and 5 mL of the Internal standard solution into a 25-mL volumetric flask and dilute with mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 354 nm Column: 4.6-mm 25-cm; packing L3 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution, Solvent blank solution, and Standard solution [NOTEThe relative retention times for anthralin, danthron (if present), dianthrone (if present), and o-nitroaniline are 1.0, 1.2, 1.7, and 2.3, respectively.] Suitability requirements Resolution: NLT 1.3, System suitability solution Tailing factor: NMT 1.5, System suitability solution Relative standard deviation: NMT 2.0% of the ratio of the peak responses, Standard solution [NOTEChromatograph Solvent blank solution: no effect on the baseline is discernible at the retention time of anthralin.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H10O3 in the portion taken: Result = (RU/RS) (CS/CU) 100 RU = response ratio of the anthralin peak to the onitroaniline peak from the Sample solution
Sample solution: 10 mg/mL of Antazoline Phosphate in methanol Identification solution: 50 g/mL from Sample solution in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Methanol, diethylamine, and ethyl acetate (2:1:17) Analysis Samples: Each Standard solution, Sample solution, and Identification solution Proceed as directed in the chapter. Position the plate in a chromatographic chamber, and develop the chromatograms in a solvent system, until the solvent front has moved about three-fourths of the length of the plate. Examine the plate under short-wavelength UV light, and compare the intensities of any secondary spots observed in the chromatogram of the Sample solution with those of the principal spots from the Standard solutions. [NOTE Disregard any spots observed at the origins of the chromatograms.] Acceptance criteria: No secondary spot from the Sample solution is larger or more intense than the principal spot of Standard solution A (0.5%), and the sum of the intensities of all secondary spots of the Sample solution corresponds to NMT 1.0%. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class Ia 741: 194198, with decomposition PH 791: 4.05.0, in a solution (1 in 50) LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Antazoline Phosphate RS
Anthralin
(Comment on this Monograph)id=m4970=Anthralin=AMonos.pdf)
C14H10O3 9(10H)-Anthracenone, 1,8-dihydroxy-; 1,8-Dihydroxy-9-anthrone [1143-38-0; 480-22-8].
226.23
DEFINITION Anthralin contains NLT 97.0% and NMT 102.0% of C14H10O3, calculated on the dried basis.
USP 32
= response ratio of the anthralin peak to the onitroaniline peak from the Standard solution = concentration of USP Anthralin RS in the CS Standard solution (g/mL) = nominal concentration of Anthralin in the Sample CU solution (g/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: RS
Official Monographs / Anthralin 249

Standard stock solution: 0.25 mg/mL of USP Anthralin RS in dichloromethane Standard solution: Standard stock solution, Internal standard solution, and Mobile phase (2:2:21) Sample solution: 5 g of Cream in a tared 100-mL beaker. Add 20 mL of dichloromethane and 10 mL of glacial acetic acid, and stir to disperse the Cream. Transfer the contents of the beaker to a filter paper (Whatman No. 4, or equivalent) with the aid of dichloromethane, and filter into a 100-mL volumetric flask. Thoroughly wash the precipitate with dichloromethane, and allow the washings to drain into the flask. Dilute with dichloromethane to volume. Pipet a volume of this solution, equivalent to 0.5 mg of anthralin, and 2 mL of Internal standard solution into a 25mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 354 nm Column: 4.6-mm 25-cm; packing L3 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution, Solvent blank solution, and Standard solution [NOTEThe relative retention times for anthralin, danthron (if present), dianthrone (if present), and o-nitroaniline are about 1.0, 1.2, 1.7, and 2.3, respectively.] Suitability requirements Resolution: NLT 1.3, System suitability solution Tailing factor: NMT 1.5, System suitability solution Relative standard deviation: NMT 2.0% of the ratio of the peak responses, Standard solution [NOTEChromatograph Solvent blank solution: no effect on the baseline is discernible at the retention time of anthralin.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H10O3 in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 = response ratio of the anthralin peak to the onitroaniline peak from the Sample solution RS = response ratio of the anthralin peak to the onitroaniline peak from the Standard solution CS = concentration of USP Anthralin RS in the Standard solution (g/mL) CU = nominal concentration of anthralin in the Sample solution (g/mL) Acceptance criteria: Cream labeled to contain more than 0.1% of anthralin contains 90.0%115.0% of the labeled amount of C14H10O3; and Cream labeled to contain 0.1% or less of anthralin contains 90.0%130.0% of the labeled amount of C14H10O3. RU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, in a cool place. Protect from light. LABELING: Label it to indicate whether the cream vehicle is aqueous or oily. USP REFERENCE STANDARDS 11 USP Anthralin RS
NMT 0.1%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 178181 LOSS ON DRYING 731: Dry Anthralin over silica gel for 4 h: it loses NMT 0.5% of its weight. ACIDITY OR ALKALINITY: Suspend Anthralin in water, and filter: the filtrate is neutral to litmus. CHLORIDE AND SULFATE, Chloride 221: Add 1 g of Anthralin to 15 mL of water, mix, and filter. Acidify 5 mL of the filtrate with nitric acid, and add a few drops of silver nitrate TS: no more opalescence is produced immediately than is present in a 5-mL portion of the filtrate to which nothing has been added. CHLORIDE AND SULFATE, Sulfate 221: To 5 mL of the untreated filtrate obtained in the test for Chloride add 3 drops of 3 N hydrochloric acid and 5 drops of barium chloride TS: no more turbidity is produced than is present in a 5-mL portion of the filtrate to which nothing has been added. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers in a cool place. Protect from light. USP REFERENCE STANDARDS 11 USP Anthralin RS
Anthralin Cream
(Comment on this Monograph)id=m4975=Anthralin Cream=AMonos.pdf) DEFINITION Anthralin Cream is Anthralin in an aqueous (oil-in-water) or oily (water-in-oil) cream vehicle. Anthralin Cream labeled to contain more than 0.1% of anthralin contains NLT 90.0% and NMT 115.0% of the labeled amount of C14H10O3; and Cream labeled to contain 0.1% or less of anthralin contains NLT 90.0% and NMT 130.0% of the labeled amount of C14H10O3. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: n-Hexane, dichloromethane, and glacial acetic acid (41:6:3) Internal standard solution: 500 g/mL of o-nitroaniline in n-hexane [NOTEFirst dissolve o-nitroaniline in a small quantity of dichloromethane, and then dilute with nhexane.] System suitability stock solution: 0.1 mg/mL of USP Anthralin RS and 0.2 mg/mL of danthron in dichloromethane System suitability solution: System suitability stock solution, n-hexane, and Mobile phase (1:1:3) Solvent blank solution: Mobile phase, n-hexane, and dichloromethane (3:1:1)
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RU
USP 32
= response ratios from the anthralin peak to the onitroaniline peak from the Sample solution RS = response ratios from the anthralin peak to the onitroaniline peak from the Standard solution CS = concentration of USP Anthralin RS in the Standard solution (g/mL) = nominal concentration of anthralin in the Sample CU solution (g/mg) Acceptance criteria: Ointment labeled to contain more than 0.1% of anthralin contains 90.0%115.0% of the labeled amount of C14H10O3; and Ointment labeled to contain 0.1% or less of anthralin contains 90.0%130.0% of the labeled amount of C14H10O3. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, in a cool place. Protect from light. USP REFERENCE STANDARDS 11 USP Anthralin RS
Anthralin Ointment
(Comment on this Monograph)id=m5000=Anthralin Ointment=A-Monos.pdf) DEFINITION Anthralin Ointment is Anthralin in a petrolatum or other oleaginous vehicle. Anthralin Ointment labeled to contain more than 0.1% of anthralin contains NLT 90.0% and NMT 115.0% of the labeled amount of C14H10O3; and Ointment labeled to contain 0.1% or less of anthralin contains NLT 90.0% and NMT 130.0% of the labeled amount of C14H10O3. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: n-Hexane, dichloromethane, and glacial acetic acid (41:6:3) Internal standard solution: 500 g/mL of o-nitroaniline in n-hexane [NOTEFirst dissolve o-nitroaniline in a small quantity of dichloromethane, and then dilute with nhexane.] System suitability stock solution: 0.1 mg/mL of USP Anthralin RS and 0.2 mg/mL of danthron in dichloromethane System suitability solution: System suitability stock solution, n-hexane, and Mobile phase (1:1:3) Solvent blank solution: Mobile phase, n-hexane, and dichloromethane (3:1:1) Standard stock solution: 0.25 mg/mL of USP Anthralin RS in dichloromethane Standard solution: Standard stock solution, Internal standard solution, and Mobile phase (2:2:21) Sample solution: 5 g of Ointment in a tared 100-mL beaker. Add 20 mL of dichloromethane and 10 mL of glacial acetic acid, and stir to disperse the Ointment. Transfer the contents of the beaker to a filter paper (Whatman No. 4, or equivalent) with the aid of dichloromethane, and filter into a 100-mL volumetric flask. Thoroughly wash the precipitate with dichloromethane, and allow the washings to drain into the flask. Dilute with dichloromethane to volume. Pipet a volume of this solution, equivalent to 0.5 mg of anthralin, and 2 mL of Internal standard solution into a 25-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 354 nm Column: 4.6-mm 25-cm; packing L3 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution, Solvent blank solution, and Standard solution [NOTEThe relative retention times for anthralin, danthron (if present), dianthrone (if present), and o-nitroaniline are 1.0, about 1.2, about 1.7, and about 2.3, respectively.] Suitability requirements Resolution: NLT 1.3, System suitability solution Tailing factor: NMT 1.5, System suitability solution Relative standard deviation: NMT 2.0% of the ratio of the peak responses , Standard solution [NOTEChromatograph Solvent blank solution: no effect on the baseline is discernible at the retention time of anthralin.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H10O3 in the portion of Ointment taken: Result = (RU/RS) (CS/CU) 100
Anthrax Vaccine Adsorbed

(Comment on this Monograph)id=m5010=Anthrax Vaccine Adsorbed=A-Monos.pdf) DEFINITION Anthrax Vaccine Adsorbed is a sterile, milky-white suspension made from cell-free filtrates of microaerophilic cultures of an avirulent, nonencapsulated strain of Bacillus anthracis. The final product contains no dead or live bacteria. The production cultures are grown in a chemically defined protein-free medium containing amino acids, vitamins, inorganic salts, and sugars. The sterile filtrate is adsorbed on sterile aluminum hydroxide, concentrated 10-fold, and resuspended in sterile physiological saline containing formaldehyde with benzethonium chloride as a preservative. Sublots may be combined to produce final lots. The product meets potency requirements when tested against the U.S. Reference Standard Anthrax Vaccine, in accordance with approved procedures (guinea pig intracutaneous challenge models). IDENTIFICATION [NOTEPerform analyses on the filtration.] PROCEDURE Trichloroacetic acid solution: Prepare a solution of trichloroacetic acid (see Reagents, Indicators, and Solutions Reagent Specifications) in water containing 100 g trichloroacetic acid per 100 mL of the solution. Sample buffer: Prepare a solution containing 141 mM tris(hydroxymethyl)aminomethane, 106 mM tris(hydroxymethyl)aminomethane hydrochloride, 0.51 mM edetate disodium, 2% (w/v) dodecyl lithium sulfate, 10% glycerol, 0.22 mM Coomassie blue G-250, and 0.175 mM phenolsulfonphthalein. If necessary, adjust with hydrochloric acid or sodium hydroxide to a pH of 8.5. Running buffer: Prepare a solution containing 25 mM tris(hydroxymethyl)aminomethane, 192 mM glycine, and 0.1% (w/v) dodecyl sodium sulfate (see Reagents, Indicators, and SolutionsReagent Specifications) in water. If necessary, adjust with hydrochloric acid or sodium hydroxide to a pH of 8.5. Transblotting buffer: Prepare a solution containing 12.5 mM tris(hydroxymethyl)aminomethane, 96 mM glycine, and 10% methanol. If necessary, adjust with hydrochloric acid or sodium hydroxide to a pH of 8.0. Blocking buffer: Prepare a solution containing 10 mM monobasic sodium phosphate, 150 mM sodium chloride, 5% (w/v) nonfat dry milk, and 0.05% (w/v) Polysorbate 20. Adjust with sodium hydroxide to a pH of 7.4.
USP 32
Primary antibody solutions: Prepare suitable monoclonal antibodies raised against the Protective Antigen (PA), the Lethal Factor (LF), and the Edema Factor (EF), respectively, of Bacillus anthracis in murine ascites cells, harvested, and used without further purification. Immediately before use, dilute each of the murine ascites fluids containing the monoclonal antibodies 1:1000 with the Blocking buffer. Secondary antibody solution: Immediately before use, dissolve according to the manufacturers instructions, if necessary, and dilute the stock horseradish peroxidase conjugated to goat anti-mouse IgG solution 1:1000 with Blocking buffer. Chromogenic visualization solution: 150 mg/mL of 4chloro-1-naphthol in water. Sample solution: Use Anthrax Vaccine Filtrate as is. Analysis: In a suitable centrifuge tube transfer 30/c mL of the Sample solution, where c is the total protein concentration, in g/mL, of the solution as determined in the test for Total protein. Add 16.5/c mL of Trichloroacetic acid solution, and incubate for at least 10 min. Centrifuge at 9000 g for 10 min, decant off the supernatant, and hold the tube inverted to drain on a filter paper. Dissolve the pellet in 60 L of Sample buffer, and transfer the solution to a polypropylene microfuge tube that has a lid. Close the lid tightly, secure with a lid-lock, and heat at 100 for 5 min. Allow the solution to cool to room temperature, and centrifuge at 10,000 g for 15 s to collect the liquids. In a suitable device for polyacrylamide-gel electrophoresis (see Electrophoresis 726 and Biotechnology-Derived Articles Polyacrylamide Gel Electrophoresis 1056) add appropriate volumes of the Running buffer in the upper and the lower buffer chambers. Attach a 4%20% gradient tris-glycine polyacrylamide slab gel sandwiched between two glass plates, such that the wells for sample application are exposed to the Running buffer in the upper buffer chamber. Apply about 20-L aliquots of the treated Sample solution in three alternate lanes. [NOTEDo not apply any solution in the outside lanes. ] Connect the lower buffer chamber electrode to the positive terminal and the upper buffer chamber electrode to the negative terminal of a suitable power supply unit, and carry out the electrophoresis at a constant current of about 40 mA. When the dye-front is about 1 cm from the bottom of the gel (about 40 min), stop the current, and remove the gel from the gel assembly. [NOTEDo not touch the gel with bare hands. Use gloves.] Place three to four filter papers, cut to the size of the gel and soaked in the Transblotting buffer, on the anode plate of a suitable semidry electroblotter. Cut a nitrocellulose membrane to the same size as the gel plus 12 mm on each side, and wet the membrane by immersing it into the Transblotting buffer for about 15 s, such that there is no air-bubble between the buffer and the membrane. Place the wet membrane immediately on the stack of filter papers, and remove all air bubbles between the membrane and filter paper by rolling a pipet, or equivalent, gently over the surface of the membrane. Place a few drops of the Transblotting buffer on the membrane, and then carefully place the gel on it. Gently roll a pipet, or equivalent, over the surface of the gel to ensure intimate contact between the gel and the membrane, making sure that there are no air bubbles in between. Place a filter paper cut to the size of the gel and soaked in the Transblotting buffer, such that there is no air-bubble between the filter paper and the gel. Place two to three additional filter papers, prepared in a similar manner, on the top, and complete the transfer stack by placing the cathode plate on the top. Apply a current of about 250 mA, and continue transfer for 90 min. Remove the membrane, and wash it quickly by immersing into water for 15 s. [NOTEDo not touch the membrane with bare hands. Use gloves.] Cut the membrane into three strips such that each strip contains a lane containing the
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Sample solution, and mark the strips as PA, LF, and EF at the top. Place each strip in a heat-sealable bag, add 5 mL of Blocking buffer, and seal the bag. Incubate for 30 min with constant agitation. Open each bag, and pour out the Blocking buffer. Add 9 mL of the diluted Primary antibody solution against PA to the bag containing the strip marked PA. Similarly, add 9 mL of the diluted Primary antibody solution against LF and EF to the bags containing strips labeled LF and EF, respectively. Seal the bags, and incubate under agitation for 2 h at room temperature or overnight at 2 to 8. Remove the strips from the plastic bags, and place in separate plastic boxes. Add sufficient Blocking buffer so that each strip is completely immersed. Agitate for at least 30 min at room temperature with two changes of Blocking buffer. Remove the strips, and place each strip in a new heat-sealable plastic bag. Add 9 mL of the Secondary antibody solution to each plastic bag. Seal the bags, and incubate for 1 h at room temperature under agitation. Remove the strips from the plastic bags, and place in separate plastic boxes. Add sufficient Blocking buffer so that each strip is completely immersed. Agitate for at least 30 min at room temperature with two changes of the Blocking buffer. Transfer each strip into a new heat-sealable plastic bag, add 9 mL of Chromogenic visualization solution and 10 L of 30% hydrogen peroxide, and seal the bags. Incubate for 30 min under agitation. Transfer the strips into separate plastic boxes, and remove the excess 4-chloro-1-naphthol by incubating with water under agitation for 10 min. Visual observation indicates a strong positive band on the strip labeled PA, a faintly detectable band on the strip labeled LF, and no detectable band on the strip labeled EF. 83 kDA PROTEIN Trichloroacetic acid solution, Sample buffer, Running buffer, and Sample solution: Prepare as directed under Identification, Procedure. Staining solution: Prepare a solution of Coomassie blue G-250 having a concentration of 1.25 g/L in a mixture of water, methanol, and acetic acid (5:4:1). Protein molecular weight standard solution: Reconstitute a vial of protein molecular weight standard mixture containing proteins of molecular weights at least in the range of 14200 kDa, according to manufacturers instruction. Dilute the solution with Sample buffer such that the concentration of each protein in the solution is about 0.5 g/L. Analysis: In a suitable centrifuge tube transfer 10/c mL of the Sample solution, where c is the total protein concentration, in g/mL, of the solution as determined by the test for Total protein (see below). Add 5.5/c mL of Trichloroacetic acid solution, and incubate for at least 10 min. Centrifuge at 9000 g for about 10 min, decant off the supernatant, and hold the tube inverted to drain on a filter paper. Dissolve the pellet in 20 L of Sample buffer, and transfer the solution to a polypropylene microfuge tube with a lid. Transfer 20 L of Protein molecular weight standard solution to another polypropylene microfuge tube with a lid. Close the lids tightly, secure with lid-locks, and heat both solutions at 100 for 5 min. Allow the solutions to cool to room temperature, and centrifuge at 10,000 g for 15 s to collect the liquids. Apply the solutions to two consecutive lanes of a 4%20% gradient tris-glycine polyacrylamide slab gel [NOTEDo not apply any solution in the outside lanes.], and electrophorese as directed under Identification (see Electrophoresis 726 and Biotechnology-Derived Articles Polyacrylamide Gel Electrophoresis 1056). When the dyefront is about 1 cm from the bottom of the gel (about 40 min), stop the current, and remove the gel from the gel assembly. Soak the gel in a suitable volume of the Staining solution for at least 1 h, such that the gel is completely immersed in the Staining solution during staining. [NOTE
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Vaccine Adsorbed with respect to the corresponding U.S. Reference Standard Anthrax Vaccine is the antilog of the horizontal distance between the two parallel lines. Acceptance criteria: The relative potency of Anthrax Vaccine Adsorbed is acceptable if it is between 0.53 and 1.79, both values inclusive. OTHER COMPONENTS [NOTEPerform analysis on final product.] ALUMINUM Not a test STANDARD SOLUTIONS: Prepare as directed under Aluminum 206, Standard Preparations, except to prepare solutions containing 10, 20, 30, 40, and 50 g/mL of aluminum. Sample solution: Mix Anthrax Vaccine Adsorbed, Final Product well, and transfer 0.2 mL to a 10-mL volumetric flask. Add 0.5 mL of concentrated sulfuric acid and 0.5 mL of concentrated nitric acid, and mix gently. Incubate at room temperature for 30 min or until the solution becomes essentially clear. Dilute with water to volume. Analysis: Proceed as directed under Aluminum 206, Procedure. Plot the absorbances versus the content of aluminum, in g/mL, for the Standard solutions, and draw a best-fit straight line through the points using a linear regression model. Calculate the amount of aluminum in Anthrax Vaccine Adsorbed, in mg/mL. The aluminum concentration is between 0.8 and 1.5 mg/mL. IMPURITIES [NOTEPerform analyses on final product.] Organic Impurities PROCEDURE 1 FORMALDEHYDE: Potassium ferricyanide solution: 25 mg/mL of potassium ferricyanide Phenylhydrazine hydrochloride solution: 4 g of phenylhydrazine hydrochloride in 100 mL of absolute alcohol, add 2 mL of water Standard stock solution: To prepare a stock solution, proceed as directed. Determine the concentration of formaldehyde in percent (w/v) as follows: Sample solution: 3 mL of Formaldehyde Solution to a tared flask containing 10 mL of water, insert the stopper in the flask tightly, and accurately determine the weight of the Formaldehyde Solution taken. Slowly and quantitatively add a mixture of 50.0 mL of 1 N sodium hydroxide VS and 50 mL of hydrogen peroxide TS that has been previously neutralized to bromothymol blue TS with 1 N sodium hydroxide. Heat the contents of the flask cautiously on a steam bath for 15 min, shaking it occasionally with a rotary motion. Allow the mixture to cool, rinse the funnel and the inner wall of the flask with water, and after allowing it to stand for 30 min, add 25 drops of bromothymol blue TS. Analysis: Titrate the excess alkali with 1 N sulfuric acid VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Also make a correction based upon the acidity found in the test for Acidity under Formaldehyde Solution. Each mL of 1 N sodium hydroxide is equivalent to 30.03 mg of Formaldehyde Solution (CH2O). Acceptance criteria: 37.0%, by weight, of CH2O for bulk containers; 36.5%, by weight, of CH2O for small containers Standard solutions: Dilute the Standard stock solution in water to obtain solutions having concentrations of 0.005%, 0.01%, and 0.02% (w/v). Sample solution: Use Anthrax Vaccine Adsorbed, Final Product as is. Analysis: To suitable glass centrifuge tubes transfer 1.0 mL each of water, the Standard solutions, and the Sample solution. To each tube add 1.0 mL of Potassium ferricyanide solution, 4.0 mL of 18% (w/v) hydrochloric acid and 2.0 mL of Phenylhydrazine hydrochloride solution. Mix after each addition. Incubate for 5060 min at room temperature. Centrifuge the solutions at 10,000 g for at least 10 min, and measure absorbances of the supernatants at 540 nm using a
Use disposable gloves.] Destain the gel with a large volume of water under constant agitation with repeated changes of water until the background of the gel is completely color free. Using the molecular weights of the proteins in Protein molecular weight standard solution, identify the band corresponding to the Protective Antigen (MW about 83 kDa) in the Sample solution lane. [NOTEThis band is also the predominant band in the lane of the Sample solution.] Scan the gel, and determine the relative amount (by peak area) of the 83 kDa band by densitometry in the lane of the Sample solution. The content of 83 kDa band is NLT 35% of the total peak area. TOTAL PROTEIN Standard solution A: Prepare a solution of albumin bovine serum (see Reagents, Indicators, and SolutionsReagent Specifications) in water to obtain a known concentration of 2.0 mg/mL. Standard solutions B, C, D, and E: Dilute Standard solution A with water to obtain solutions having protein concentrations of 4, 8, 16, and 24 g/mL, respectively. Sample solution: Use Anthrax Vaccine Filtrate as is. Analysis (see Biotechnology-Derived ArticlesTotal Protein Assay 1057, Method 3): To a series of test tubes transfer 800 L each of Standard solutions B, C, D, and E and the Sample solution. Also transfer 800 L of water to be used as the blank. Add 200 L of Coomassie blue G-250 dye solution (see Reagents, Indicators, and SolutionsReagent Specifications) to each tube, and mix without foaming. Determine absorbances of the solutions at 595 nm using a suitable spectrophotometer (see Spectrophotometry and Light-Scattering 851), using the blank to set the instrument to zero. [NOTEDo not use quartz (silica) spectrophotometer cells; the dye binds to silica.] Construct a standard curve by plotting the absorbances versus protein concentrations, in g/mL, of Standard solutions B, C, D, and E and by drawing a best-fit straight line using the linear regression method. From the standard curve, determine the total protein concentration of the Sample solution using the absorbance value. The protein concentration is between 5 and 20 g/mL. ASSAY [NOTEPerform analysis on the final product.] RELATIVE POTENCY Standard solutions: Dilute approved U.S. Reference Standard Anthrax Vaccine 1:1.6, 1:4, 1:10, and 1:25 aseptically with a sterile 0.9% sodium chloride solution. Sample solutions: Dilute Anthrax Vaccine Adsorbed, Final Product 1:1.6, 1:4, 1:10, and 1:25 aseptically with a sterile 0.9% sodium chloride solution. Analysis Samples: Standard solutions and Sample solutions Assign each dilution to a set of 12 randomly selected guinea pigs, strain Mdh:S(RA), 6 males and 6 females, each weighing 315385 g on the day of vaccination. Inject the animals subcutaneously in the ventral abdomen with 0.5 mL of the assigned dilutions. On the 14th day post-vaccination, challenge the animals with approximately 1000 spores of Bacillus anthracis strain Vollum 1B, and record the deaths daily for a 10-day observation period. Record the numbers of surviving animals for each of the Standard solutions and the Sample solutions at the end of the test. Perform calculations by estimating best-fit lines for the Standard solutions and the Sample solutions using a logistic regression model that utilizes the number of animals that survived at the end of the test and the time to death for the animals that died. Evaluate statistically the lines corresponding to the Standard solutions and the Sample solutions for parallelism. Determine the common slope, and draw the parallel lines using the common slope. The relative potency of Anthrax
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suitable spectrophotometer (see Spectrophotometry and Light-Scattering 851). Plot the absorbances versus concentrations of formaldehyde, in mg/mL, in the Standard solutions, and draw the best-fit straight line through the points. Calculate the amount of formaldehyde in the sample in percent (w/v). Acceptance criteria: The concentration of formaldehyde in Anthrax Vaccine Adsorbed is less than 0.02% (w/v). PROCEDURE 2: BENZETHONIUM CHLORIDE Citrate buffer: 25 g of citric acid monohydrate in 60 mL of water, and adjust with a solution of sodium hydroxide to a pH of 4.5. Transfer the solution to a 100-mL volumetric flask. Dilute with water to volume. Dye solution: 50 mg of 2,4,5,7-tetrabromofluorescein in 100 mL of water, and mix. Dilute 1 mL of this solution with water to 100 mL. Docusate sodium solution: 50 g/mL of docusate sodium Standard solution A: 0.5 g of benzethonium chloride in a 100-mL volumetric flask, dissolve in 60 mL water, and dilute with water to volume Standard solutions B, C, D, and E: Dilute Standard solution A with water to obtain solutions having concentrations of 0.001%, 0.002%, 0.003%, and 0.004% (w/v), respectively. Sample solution: Use Anthrax Vaccine Adsorbed, Final Product as is. Analysis: Transfer 4.0 mL each of Standard solutions B, C, D, and E and the Sample solution to suitable glass centrifuge tubes. Add 1.0 mL of Citrate buffer and 0.4 mL of the Dye solution to each tube. Add 4.0 mL of 1,1,2,2tetrachloroethane to each tube, and vigorously mix on a vortex mixer for 1 min. Centrifuge at about 1000 g for at least 15 min to separate the organic layer from the aqueous layer. Transfer 2.0 mL of the organic layer from the tubes to another set of glass tubes. Add 4.0 mL of water and 0.5 mL of Citrate buffer to each tube, and mix on a vortex mixer for about 1 min. Titrate the benzethonium chloride-dye complex in each tube with the Docusate sodium solution (see Titrimetry 541) to the colorimetric endpoint indicated by the disappearance of the pink color of the organic layer. [NOTEVigorously mix the solution on a vortex mixer after each addition of the Docusate sodium solution.] Plot the volumes of Docusate sodium solution required versus the concentrations of benzethonium chloride in Standard solutions B, C, D, and E, and draw a best-fit straight line through the points. Determine the concentration of benzethonium chloride in the Sample solution from the volume of Docusate sodium solution required to titrate the Sample solution. Acceptance criteria: The concentration of benzethonium chloride in Anthrax Vaccine Adsorbed is between 0.0015% and 0.0030% (w/v). SPECIFIC TESTS [NOTEPerform tests on final product.] SAFETY: It meets the requirements when tested as directed in Biological Reactivity Tests, In Vivo 88, Safety Tests Biologicals. STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Direct Inoculation of the Culture Medium. PH 791: 7.58.5 SODIUM CHLORIDE Standard solutions A and B: Prepare two solutions of sodium chloride in water having concentrations of 0.2 mM and 2.0 mM, respectively. Sample solution: Transfer 0.5 mL of Anthrax Vaccine Adsorbed, Final Product to a 50-mL volumetric flask. Dilute with water to volume. Analysis: Determine the voltage readings of Standard solutions A and B and the Sample solution using an ionspecific electrode specific for the chloride ion electrically
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coupled with a standard silversilver chloride reference electrode. Plot the voltage readings versus concentration of chloride, in mg/mL, for Standard solutions A and B, and draw a straight line joining the points. Calculate the concentration of chloride ion in the Sample solution from the voltage reading. Assuming that the chloride ion comes entirely from sodium chloride, calculate the concentrations of sodium chloride in the Sample solution. Acceptance criteria: The concentration of sodium chloride in Anthrax Vaccine Adsorbed is between 0.75% and 0.95% (w/v). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in multiple-dose tight Type I glass containers. Store at a temperature between 2 and 8. Do not freeze. LABELING: Label it to state that it is to be well shaken before use and that it is not to be frozen. EXPIRATION DATE: The expiration date is 18 months from the date of manufacture.
Anticoagulant Citrate Dextrose Solution

(Comment on this Monograph)id=m5100=Anticoagulant Citrate Dextrose Solution=A-Monos.pdf) DEFINITION Anticoagulant Citrate Dextrose Solution is a sterile solution of Citric Acid, Sodium Citrate, and Dextrose in Water for Injection. It contains in each 1000 mL:
Solution A NLT 20.59 g NMT 22.75 g Dextrose (C6H12O6H2O) NLT 23.28 g NMT 25.73 g Sodium (Na) NLT 4.90 g NMT 5.42 g NLT 2.94 g NMT 3.25 g NLT 13.96 g NMT 15.44 g Solution B NLT 12.37 g NMT 13.67 g
Total Citrate, expressed as citric acid, anhydrous (C6H8O7)
It contains no antimicrobial agents. Prepare Anticoagulant Citrate Dextrose Solution as follows:

Solution A Citric Acid (anhydrous) Sodium Citrate (dihydrate) Dextrose (monohydrate) Water for Injection 7.3 g 22.0 g 24.5 g 1000 mL Solution B 4.4 g 13.2 g 14.7 g 1000 mL
Dissolve the ingredients, and mix. Filter the solution until clear, place immediately in suitable containers, and sterilize. If desired, 8 g and 4.8 g of monohydrated citric acid may be used instead of the indicated, respective amounts of anhydrous citric acid; 19.3 g and 11.6 g of anhydrous sodium citrate may be used instead of the indicated, respective amounts of dihydrated sodium citrate; and 22.3 g and 13.4 g of anhydrous dextrose may be used instead of the indicated, respective amounts of monohydrated dextrose. IDENTIFICATION Add a few drops of solution (1 in 20) to 5 mL of hot alkaline cupric tartrate TS: a copious red precipitate of cuprous oxide
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A
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= 100 mm divided by the length of the polarimeter tube (mm) R = observed rotation (degrees) Where the Solution is labeled to contain dextrose monohydrate, calculate the percentage (g/100 mL) C6H12O6 H2O in the portion of Solution taken: Result = (100/F) (Mr1/Mr2) A R 100 F Mr1 Mr2 A R = percentage = midpoint of the specific rotation range for anhydrous dextrose (degrees), 52.9 = molecular weight for dextrose monohydrate, 198.17 = molecular weight for anhydrous dextrose, 180.16 = 100 mm divided by the length of the polarimeter tube (mm) = observed rotation (degrees)
is formed. When concentrated to one-half its volume, it meets the requirements of the tests for Identification Tests Citrate 191 and for Identification TestsSodium 191 ASSAY TOTAL CITRATE Mobile phase, Standard solution A, and Chromatographic system: Proceed as directed under Assay for Citric Acid/Citrate and Phosphate 345. Sample solution: Pipet 5 mL of Solution into a suitable volumetric flask, and proceed as directed for Assay Preparation for Citric Acid/Citrate Assay under General Chapter 345. Analysis: Proceed as directed for Analysis under General Chapter 345. Calculate the quantity, in mg, of anhydrous citric acid (C6H8O7) in the volume of Solution taken: Result = (Mr1/Mr2) CS (rU/rS) 0.001 D Mr1 Mr2 CS = molecular weight of anhydrous citric acid, 192.12 = molecular weight of citrate (C6H5O7), 189.10 = concentration of citrate in Standard solution A (g/mL) = citrate peak area of the Sample solution = citrate peak area of Standard solution A = dilution factor
IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion shows no more chloride than corresponds to 0.50 mL of 0.020 N hydrochloric acid (0.0035%). SPECIFIC TESTS PH 791: 4.55.5 OTHER REQUIREMENTS: Meets the requirements under Injections 1 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP Endotoxin Units/mL. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, of colorless, transparent Type I or Type II glass, or of a suitable plastic material (see Transfusion and Infusion Assemblies and Similar Medical Devices 161). LABELING: Label to indicate the number of mL of Solution required/100 mL of whole blood or the number of mL of Solution required/volume of whole blood to be collected. USP REFERENCE STANDARDS 11 USP Citric Acid RS USP Endotoxin RS
rU rS D SODIUM Solution A: Add 1.04 g of lithium nitrate to a 1000-mL volumetric flask, add a suitable nonionic surfactant, and add water to volume. This solution contains 15 mEq of lithium/1000 mL. Standard solution: Add 8.18 g of sodium chloride, previously dried at 105 for 2 h to a 1000-mL volumetric flask, and dilute with water to volume. This solution contains 140 mEq of sodium/1000 mL. Transfer 50 L of this solution to a 10-mL volumetric flask, and dilute with Solution A to volume. Sample solution: Add 25 mL of Solution into a 50-mL volumetric flask, and dilute with water to volume. Transfer 50 L of this solution to a 10-mL volumetric flask, and dilute with Solution A to volume. Analysis: Using a suitable flame photometer, adjusted to read zero with Solution A, concomitantly determine the sodium flame emission readings for the Standard solution and the Sample solution at the wavelength of maximum emission at 589 nm. Calculate the quantity, in g, of Na in 1000 mL of Solution taken: Result = W (Ar/Mr) (rU/rS) W Ar Mr rU rS = weight of sodium chloride taken to make the Standard solution (g), 8.18 = atomic weight of sodium, 22.99 = molecular weight of sodium chloride, 58.44 = sodium emission readings of the Sample solution = sodium emission readings of the Standard solution
Anticoagulant Citrate Phosphate Dextrose Solution

(Comment on this Monograph)id=m5130=Anticoagulant Citrate Phosphate Dextrose Solution=A-Monos.pdf) DEFINITION Anticoagulant Citrate Phosphate Dextrose Solution is a sterile solution of Citric Acid, Sodium Citrate, Monobasic Sodium Phosphate, and Dextrose in Water for Injection. It contains, in each 1000 mL, NLT 2.11 g and NMT 2.33 g of monobasic sodium phosphate (NaH2PO4 H2O); NLT 24.22 g and NMT 26.78 g of dextrose (C6H12O6 H2O); NLT 19.16 g and NMT 21.18 g of total citrate, expressed as citric acid, anhydrous (C6H8O7); and NLT 6.21 g and NMT 6.86 g of Sodium (Na). It contains no antimicrobial agents. Prepare Anticoagulant Citrate Phosphate Dextrose Solution as follows.
Citric Acid (anhydrous) Sodium Citrate (dihydrate) Monobasic sodium phosphate (monohydrate; NaH2PO4H2O) Dextrose (monohydrate) 2.99 g 26.3 g 2.22 g 25.5 g
DEXTROSE Analysis: Determine the angular rotation of Solution in a suitable polarimeter tube (see Optical Rotation 781). Where the Solution is labeled to contain anhydrous dextrose, calculate the percentage (g/100 mL) of C6H12O6 in the portion of Solution taken: Result = (100/F) A R 100 F = percentage = midpoint of the specific rotation range for anhydrous dextrose (degrees), 52.9
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Water for Injection To make A sufficient quantity 1000 mL

the beaker. Heat the beaker and contents over a burner that has been adjusted to cause boiling of the solution to start in 3.54 min. Boil the solution for 2 min, accurately timed, and filter immediately through the tared crucible, taking care to transfer all of the boiling chips or glass beads to the crucible. Wash the precipitate with hot water and 10 mL of alcohol. Dry the crucible and contents at 110 to constant weight. Perform a blank determination, and correct the weight of the precipitate from the sample for any precipitate obtained in the blank. Each mg of cuprous oxide precipitate of the substance under assay is equivalent to 0.496 mg of C6H12O6 H2O. SODIUM Solution A: Transfer 1.04 g of lithium nitrate to a 1000-mL volumetric flask, add a suitable nonionic surfactant, and add water to volume. This solution contains 15 mEq of lithium/1000 mL. Standard solution B: Transfer 8.18 g of sodium chloride, previously dried at 105 for 2 h to a 1000-mL volumetric flask, and dilute with water to volume. This solution contains 140 mEq of sodium/1000 mL. Transfer 50 L of this solution to a 10-mL volumetric flask, and dilute with Solution A to volume. Sample solution: Transfer 25 mL of Solution into a 50-mL volumetric flask, and dilute with water to volume. Transfer 50 L of this solution to a 10-mL volumetric flask, and dilute with Solution A to volume. Analysis: Using a suitable flame photometer, adjusted to read zero with Solution A, concomitantly determine the sodium flame emission readings for the Standard solution and the Sample solution at the wavelength of maximum emission at 589 nm. Calculate the quantity, in g, of Na in 1000 mL of Solution taken: Result = (rU/rS) (Ar/Mr) W rU rS Ar Mr W = sodium emission readings from the Sample solution = sodium emission readings from the Standard solution = atomic weight of sodium, 22.99 = molecular weight of sodium chloride, 58.44 = weight of sodium chloride taken to make the Standard solution (g), 8.18
Dissolve the ingredients, and mix. Filter the solution until clear, place immediately in suitable containers, and sterilize. If desired, 3.27 g of monohydrated citric acid may be used instead of the indicated amount of anhydrous citric acid; 23.06 g of anhydrous sodium citrate may be used instead of the indicated amount of dihydrated sodium citrate; 1.93 g of anhydrous monobasic sodium phosphate may be used instead of the indicated amount of monohydrated monobasic sodium phosphate; and 23.2 g of anhydrous dextrose may be used instead of the indicated amount of monohydrated dextrose. IDENTIFICATION It meets the requirements for Identification TestsGeneral 191, Phosphates and the following test. Add a few drops of a solution (1 in 20) to 5 mL of hot alkaline cupric tartrate TS: a copious red precipitate of cuprous oxide is formed. When concentrated to one-half its volume, meets the requirements for Identification TestsGeneral 191, Citrate and 191, Sodium. ASSAY TOTAL CITRATE AND TOTAL PHOSPHATE Mobile phase, Standard solution B, and Chromatographic system: Proceed as directed under Assay for Citric Acid/Citrate and Phosphate 345. Sample solution for total citrate: Pipet 10 mL of Solution into a suitable volumetric flask, and proceed as directed for Assay for Citric Acid/Citrate and Phosphate 345, Assay Preparation for Citric Acid/Citrate Assay. Sample solution for total phosphate: Pipet 5 mL of Solution into a suitable volumetric flask, and proceed as directed for Assay for Citric Acid/Citrate and Phosphate 345, Assay Preparation for Phosphate Assay. Analysis: Proceed as directed for Assay for Citric Acid/Citrate and Phosphate 345, Procedure. Calculate the quantity, in mg, of C6H8O7 in the volume of Solution taken: Result = (rU/rS) CS (Mr1/Mr2) 0.001 D = citrate peak area from the Sample solution for total citrate = citrate peak area from Standard solution B rS = concentration of citrate in Standard solution B CS (g/mL) Mr1 = molecular weight of anhydrous citric acid, 192.12 Mr2 = molecular weight of citrate (C6H5O7), 189.10 D = dilution factor Calculate the quantity of phosphate, in mg, expressed as NaH2PO4 H2O, in the volume of Solution taken: rU Result = (rU/rS) CS (Mr1/Mr2) rU rS CS Mr1 = phosphate peak area from the Sample solution for total phosphate = phosphate peak area from Standard solution B = concentration of phosphate in Standard solution B (g/mL) = molecular weight of monobasic sodium phosphate monohydrate, 137.99 = molecular weight of phosphate (PO4), 94.97
IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion shows no more chloride than corresponds to 0.50 mL of 0.020 N hydrochloric acid (0.0035%). SPECIFIC TESTS PH 791: 5.06.0 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP Endotoxin Units/mL. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, of colorless, transparent, Type I or Type II glass, or of a suitable plastic material (see Transfusion and Infusion Assemblies and Similar Medical Devices 161). LABELING: Label it to indicate the number of mL of Solution required/100 mL of whole blood or the number of mL of Solution required/volume of whole blood to be collected. USP REFERENCE STANDARDS 11 USP Citric Acid RS USP Endotoxin RS
Mr2 DEXTROSE Tare a clean, medium-porosity filtering crucible containing several carborundum boiling chips or glass beads. Pipet 50 mL of freshly mixed alkaline cupric tartrate TS into a 400mL beaker. Add the boiling chips or glass beads from the tared crucible, 45 mL of water, and 5.0 mL of Solution to
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Calculate the quantity of phosphate, in mg, expressed as NaH2PO4 H2O, in the volume of Solution taken: Result = (rU/rS) CS (Mr1/Mr2) 0.001 D rU rS CS Mr1 = phosphate peak area from the Sample solution for total phosphate = phosphate peak area from Standard solution B = concentration of phosphate in Standard solution B (g/mL) = molecular weight of monobasic sodium phosphate monohydrate, 137.99 = molecular weight of phosphate (PO4), 94.97 = dilution factor
Anticoagulant Citrate Phosphate Dextrose Adenine Solution

(Comment on this Monograph)id=m5140=Anticoagulant Citrate Phosphate Dextrose Adenine Solution=A-Monos.pdf) DEFINITION Anticoagulant Citrate Phosphate Dextrose Adenine Solution is a sterile solution of Citric Acid, Sodium Citrate, Monobasic Sodium Phosphate, Dextrose, and Adenine in Water for Injection. It contains, in each 1000 mL, NLT 2.11 g and NMT 2.33 g of monobasic sodium phosphate (NaH2PO4 H2O); NLT 30.30 g and NMT 33.50 g of dextrose (C6H12O6 H2O); NLT 19.16 g and NMT 21.18 g of total citrate, expressed as citric acid, anhydrous (C6H8O7); NLT 6.21 g and NMT 6.86 g of sodium (Na); and NLT 0.247 g and NMT 0.303 g of adenine (C5H5N5). It contains no antimicrobial agents. Prepare Anticoagulant Citrate Phosphate Dextrose Adenine Solution as follows.
Citric Acid (anhydrous) Sodium Citrate (dihydrate) Monobasic Sodium Phosphate (monohydrate; NaH2PO4H2O) Dextrose (monohydrate) C5H5N5 Water for Injection To make 2.99 g 26.3 g 2.22 g 31.9 g 0.275 g A sufficient quantity 1000 mL
Dissolve the ingredients, and mix. Filter the solution until clear, place immediately in suitable containers, and sterilize. If desired, 3.27 g of monohydrated citric acid may be used instead of the indicated amount of anhydrous citric acid; 23.06 g of anhydrous sodium citrate may be used instead of the indicated amount of dihydrated sodium citrate; 1.93 g of anhydrous monobasic sodium phosphate may be used instead of the indicated amount of monohydrated monobasic sodium phosphate; and 29.0 g of anhydrous dextrose may be used instead of the indicated amount of monohydrated dextrose. ASSAY TOTAL CITRATE AND TOTAL PHOSPHATE Mobile phase, Standard solution B, and Chromatographic system: Proceed as directed under Assay for Citric Acid/Citrate and Phosphate 345. Sample solution for total citrate: Pipet 10 mL of Solution into a suitable volumetric flask, and proceed as directed for Assay for Citric Acid/Citrate and Phosphate 345, Assay Preparation for Citric Acid/Citrate Assay. Sample solution for total phosphate: Pipet 5 mL of Solution into a suitable volumetric flask, and proceed as directed for under Assay for Citric Acid/Citrate and Phosphate 345, Assay Preparation for Phosphate Assay. Analysis: Proceed as directed for Assay for Citric Acid/Citrate and Phosphate 345, Procedure. Calculate the quantity, in mg, of C6H8O7 in the volume of Solution taken: Result = (rU/rS) CS (Mr1/Mr2) 0.001 D rU rS CS Mr1 Mr2 D = citrate peak area from the Sample solution for total citrate = citrate peak area from Standard solution B = concentration of citrate in Standard solution B (g/mL) = molecular weight of anhydrous citric acid, 192.12 = molecular weight of citrate (C6H5O7), 189.10 = dilution factor
Mr2 D SODIUM Solution A: Transfer 1.04 g of lithium nitrate to a 1000-mL volumetric flask, add a suitable nonionic surfactant, and add water to volume. This solution contains 15 mEq of lithium/1000 mL. Standard solution: Transfer 8.18 g of sodium chloride, previously dried at 105 for 2 h to a 1000-mL volumetric flask, and dilute with water to volume. This solution contains 140 mEq of sodium/1000 mL. Transfer 50 L of this solution to a 10-mL volumetric flask, and dilute with Solution A to volume. Sample solution: Transfer 25 mL of Solution into a 50-mL volumetric flask, and dilute with water to volume. Transfer 50 L of this solution to a 10-mL volumetric flask, and dilute with Solution A to volume. Analysis: Using a suitable flame photometer, adjusted to read zero with Solution A, concomitantly determine the sodium flame emission readings for the Standard solution and the Sample solution at the wavelength of maximum emission at 589 nm. Calculate the quantity, in g, of Na in 1000 mL of Solution taken: Result = (rU/rS) (Ar/Mr) W 2 rU rS Ar Mr W = sodium emission readings from the Sample solution = sodium emission readings from the Standard solution = atomic weight of sodium, 22.99 = molecular weight of sodium chloride, 58.44 = weight of sodium chloride taken to make the Standard solution (g), 8.18
DEXTROSE Tare a clean, medium-porosity filtering crucible containing several carborundum boiling chips or glass beads. Pipet 50 mL of freshly mixed alkaline cupric tartrate TS into a 400mL beaker. Add the boiling chips or glass beads from the tared crucible, 45 mL of water, and 5.0 mL of Solution to the beaker. Heat the beaker and contents over a burner that has been adjusted to cause boiling of the solution to start in 3.54 min. Boil the solution for 2 min, accurately timed, and filter immediately through the tared crucible, taking care to transfer all of the boiling chips or glass beads to the crucible. Wash the precipitate with hot water and 10 mL of alcohol. Dry the crucible and contents at 110 to constant weight. Perform a blank determination, and make any necessary correction. Each mg of cuprous oxide precipitate obtained is equivalent to 0.496 mg of C6H12O6 H2O. ADENINE Mobile phase: To 3.45 g of ammonium dihydrogen phosphate in 950 mL of water, add 10 mL of glacial acetic acid, and dilute with water to 1000 mL. Standard solutions: Quantities of USP Adenine RS in dilute hydrochloric acid (1 in 120) in three separate volumetric
USP 32
flasks, dilute with the dilute hydrochloric acid solution to volume to obtain Standard solutions having known concentrations of 0.25, 0.275, and 0.30 mg of adenine/mL, respectively. Protect from light. System suitability solution: 0.275 mg/mL of each USP Adenine RS and purine, in dilute hydrochloric acid (1 in 120) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm stainless steel; packing L9 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: System suitability solution (NLT four injections) Suitability requirements Resolution: NLT 3.0 between adenine and purine Relative standard deviation: NMT 2.5% for adenine peak and NMT 2.0% for the retention time of adenine peak Analysis Samples: Standard solution and Sample solution Plot the responses against the concentrations, in mg, of USP Adenine RS/mL of the Standard solutions. Calculate the quantity, in mg, of C5H5N5 in each mL of the Solution taken as the value read directly from the Standard curve corresponding to the response obtained from the portion of the Solution chromatographed. IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion shows no more chloride than corresponds to 0.50 mL of 0.020 N hydrochloric acid (0.0035%). SPECIFIC TESTS PH 791: 5.06.0 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP Endotoxin Units/mL. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, of colorless, transparent, Type I or Type II glass, or of a suitable plastic material (see Transfusion and Infusion Assemblies and Similar Medical Devices 161). LABELING: Label it to indicate the number of mL of solution required/100 mL of whole blood or the number of mL of solution required/volume of whole blood to be collected. USP REFERENCE STANDARDS 11 USP Adenine RS USP Citric Acid RS USP Endotoxin RS

Heparin Sodium Sodium Chloride Injection To make 75,000 Units A sufficient quantity 1000 mL
Add the Heparin Sodium, in solid form or in solution, to the Sodium Chloride Injection, mix, filter if necessary, and sterilize. ASSAY HEPARIN SODIUM Standard solution: Determine by preliminary trial, if necessary, approximately the minimum quantity of USP Heparin Sodium RS which, when added in 0.8 mL of saline TS, maintains fluidity in 1 mL of prepared plasma for 1 h after the addition of 0.2 mL of calcium chloride solution (1 in 100). This quantity is usually between 1 and 3 USP Heparin Units. On the day of the assay prepare a Standard solution such that it contains, in each 0.8 mL of saline TS, the above-determined quantity of the Reference Standard. Sample solution: Dilute the Solution in sufficient saline TS to give a concentration estimated to correspond to that of the Standard solution. Preparation of plasma: Collect blood from sheep directly into a vessel containing 8% sodium citrate solution in the proportion of one volume to each 19 volumes of blood to be collected. Mix immediately by gentle agitation and inversion of the vessel. Promptly centrifuge the blood, and pool the separated plasma. To a 1-mL portion of the pooled plasma in a clean test tube, add 0.2 mL of calcium chloride solution (1 in 100). Consider the plasma suitable for use if a solid clot forms within 5 min. To store plasma for future use, subdivide the pooled lot into portions not exceeding 100 mL in volume, and store in the frozen state, preventing even partial thawing prior to use. For use in the assay, thaw the frozen plasma in a water bath at a temperature not exceeding 37. Remove particulate matter by straining the thawed plasma through a coarse filter. Analysis Samples: Standard solution and Sample solution To meticulously clean 13-mm 100-mm test tubes, add graded amounts of the Standard solution, selecting the amounts so that the largest does not exceed 0.8 mL, and so that they correspond roughly to a geometric series in which each step is approximately 5% greater than the next lower. To each tube so prepared, add sufficient saline TS to make the total volume 0.8 mL. Add 1.0 mL of prepared plasma to each tube. Then add 0.2 mL of calcium chloride solution (1 in 100), note the time, immediately insert a suitable stopper in each tube, and mix the contents by inverting three times in such a way that the entire inner surface of the tube is wet. In the same manner set up a series using the Sample solution, completing the entire process of preparing and mixing the tubes of both the Standard solution and the Sample solution within 20 min after the addition of the prepared plasma. In one h, accurately timed, after the addition of the calcium chloride, determine the extent of clotting in each tube, recognizing three grades (0.25, 0.50, and 0.75) between zero and full clotting (1.0). If the series does not contain two tubes graded more than 0.5 and two tubes graded less than 0.5, repeat the Assay, using appropriately modified Standard solution and Sample solution. Calculation: Convert to logarithms the volumes of Standard solution used in the successive five or six tubes that bracket a grade of clotting of 0.5, including at least two tubes with a larger and two tubes with a smaller grade than 0.5. Number and list the tubes serially, and tabulate for each the grade of clotting observed in each tube. From the log-
Anticoagulant Heparin Solution

(Comment on this Monograph)id=m5190=Anticoagulant Heparin Solution=A-Monos.pdf) DEFINITION Anticoagulant Heparin Solution is a sterile solution of Heparin Sodium in Sodium Chloride Injection. Its potency is NLT 90.0% and NMT 110.0% of the potency stated on the label in terms of USP Heparin Units. It contains NLT 0.85% and NMT 0.95% of sodium chloride (NaCl). It may be buffered. It contains no antimicrobial agents. Prepare Heparin solution as follows:
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Sodium Citrate (dihydrate) Water for Injection To make 40 g
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volumes, x, and separately from their corresponding grades of clotting, y, compute the paired averages xi and yi of Tubes 1, 2, and 3; of Tubes 2, 3, and 4; of Tubes 3, 4, and 5; and, where the series consists of 6 tubes, of Tubes 4, 5, and 6, respectively. If for one of these paired averages the average grade, yi, is exactly 0.50, the corresponding xi is the median log-volume of the Standard solution, xs. Otherwise, interpolate xs from the paired values of yi, xi and yi +1, xi +1 that fall immediately below and above grade 0.5 as: xS = xi + (yi 0.5)(xi +1 xi)/(yi yi+1) From the paired data on the tubes of the Sample solution, compute similarly its median log-volume u. The log potency of the Sample solution is: M = xS xU + log R where R = vS/vU is the ratio of the USP Heparin Units (vS)/mL of the Standard solution to the mg (vU) of Anticoagulant Heparin Solution/mL of the Sample solution. Repeat the assay independently, and average the two or more values of M to obtain M. If the second determination of M differs by more than 0.05 from the first determination, continue the Assay until the log confidence interval computed as directed under Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays does not exceed 0.20. The potency of Solution in USP Heparin Units/mL is P* = antilog M. Acceptance criteria: 90.0%110.0% SODIUM CHLORIDE Sample solution: Solution and potassium chromate TS (5:1) Analysis: Titrate with 0.1 N silver nitrate VS. Each mL of 0.1 N silver nitrate is equivalent to 5.844 mg of NaCl. Acceptance criteria: 0.85%0.95% SPECIFIC TESTS PH 791: 5.07.5 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 2.5 USP Endotoxin Units/mL. INJECTIONS 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, of colorless, transparent Type I or Type II glass, or of a suitable plastic material (see Transfusion and Infusion Assemblies and Similar Medical Devices 161). LABELING: Label it in terms of USP Heparin Units, and to indicate the number of mL of Solution required per 100 mL of whole blood. USP REFERENCE STANDARDS 11 USP Endotoxin RS USP Heparin Sodium RS
A sufficient quantity 1000 mL
[NOTEAnhydrous sodium citrate (35.1 g) may be used instead
of the dihydrate.] Dissolve the Sodium Citrate in sufficient Water for Injection to make 1000 mL, and filter until clear. Place the solution in suitable containers, and sterilize.
IDENTIFICATION IDENTIFICATION TESTSGENERAL, Sodium 191 and Citrate 191: When evaporated to a concentration of 1 in 20, it meets the requirements. ASSAY PROCEDURE Mobile phase, Standard solution A, and Chromatographic system: Proceed as directed under Assay for Citric Acid/Citrate and Phosphate 345. Sample solution: Pipet 10 mL of Solution into a suitable volumetric flask, and proceed as directed for Assay Preparation for Citric Acid/Citrate Assay under General Chapter 345. Analysis: Proceed as directed for Analysis under General Chapter 345. Calculate the quantity, in mg, of C6H5Na3O7 2H2O in the volume of Solution taken: Result = CS (rU/rS) (Mr1/Mr2) D 0.001 = concentration of citrate in Standard solution A (g/mL) rU = citrate peak area of the Sample solution rS = citrate peak area of Standard solution A Mr1 = molecular weight of anhydrous citric acid, 294.10 = molecular weight of citrate (C6H5O7), 189.10 Mr2 D = dilution factor Acceptance criteria: 3.80 g4.20 g SPECIFIC TESTS PH 791: 6.47.5 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP Endotoxin Units/mL. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I or Type II glass. USP REFERENCE STANDARDS 11 USP Citric Acid RS USP Endotoxin RS CS
Anticoagulant Sodium Citrate Solution

(Comment on this Monograph)id=m5220=Anticoagulant Sodium Citrate Solution=A-Monos.pdf) DEFINITION Anticoagulant Sodium Citrate Solution is a sterile solution of Sodium Citrate in Water for Injection. It contains, in each 100 mL, NLT 3.80 g and NMT 4.20 g of sodium citrate dihydrate (C6H5Na3O7 2H2O). It contains no antimicrobial agents.
Antihemophilic Factor
(Comment on this Monograph)id=m5250=Antihemophilic Factor=A-Monos.pdf) DEFINITION Antihemophilic Factor conforms to the regulations of the FDA concerning biologics (see Biologics 1041). It is a sterile, freeze-dried powder containing the Factor VIII fraction
USP 32
prepared from units of human venous plasma that have been tested for the absence of hepatitis B surface antigen, obtained from whole-blood donors and pooled. It may contain Heparin Sodium or Sodium Citrate. It meets the requirements of the test for potency, by comparison with the U.S. Standard Antihemophilic Factor (Factor VIII) or with a working reference that has been calibrated with it, in containing NLT 80% and NMT 120% of the potency stated on the label, the stated potency being NLT 100 Antihemophilic Factor Units/g of protein. It meets the requirements of the test for pyrogen, the test dose being 10 Antihemophilic Factor Units/kg. SPECIFIC TESTS EXPIRATION DATE: The expiration date is not later than 2 years from date of manufacture, within which time it may be stored at room temperature and used within 6 months of the time of such storage. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in hermetic containers, in a refrigerator, unless otherwise indicated. LABELING: Label it to state that it is to be used within 4 h after constitution, that it is for intravenous administration, and that a filter is to be used in the administration equipment.
Official Monographs / Antimony 259
Antimony Potassium Tartrate

(Comment on this Monograph)id=m5290=Antimony Potassium Tartrate=A-Monos.pdf)
667.87 C8H4K2O12Sb2 3H2O Antimonate(2-), bis[-[2,3-dihydroxybutanedioato(4-)O1,O2:O3,O4]]-di-, dipotassium, trihydrate, stereoisomer; Dipotassium bis[-[L-(+)-tartrato(4-)]]diantimonate(2-) trihydrate [28300-74-5]. Anhydrous 613.82 [11071-15-1]. DEFINITION Antimony Potassium Tartrate contains NLT 99.0% and NMT 103.0% of C8H4K2O12Sb2 3H2O. IDENTIFICATION A. When heated to redness, it chars, emits an odor resembling that of burning sugar, and leaves a blackened residue. This residue has an alkaline reaction, and when a small fragment of it is held in a nonluminous flame, the flame is tinted violet. B. In a solution (1 in 20), acidified with hydrochloric acid, hydrogen sulfide TS produces an orange-red precipitate, which is soluble in ammonium sulfide TS and in 1 N sodium hydroxide. C. IDENTIFICATION TESTSGENERAL 191, Tartrate ASSAY PROCEDURE Sample: 500 mg of Antimony Potassium Tartrate Analysis: Dissolve in 50 mL of water, add 5 g of potassium sodium tartrate, 2 g of sodium borate, and 3 mL of starch TS, and immediately titrate with 0.1 N iodine VS to the production of a persistent blue color. Each mL of 0.1 N iodine is equivalent to 16.70 mg of C8H4K2O12Sb2 3H2O. Acceptance criteria: 99.0%103.0% IMPURITIES Inorganic Impurities ARSENIC 211 Sample solution: Dissolve 100 mg in 5 mL of hydrochloric acid. Add 10 mL of a recently prepared solution of 20 g of stannous chloride in 30 mL of hydrochloric acid. Analysis: Transfer the Sample solution to a colorcomparison tube, and allow to stand for 30 min. Viewed downward over a white surface, the color of the solution appears no deeper than that of a blank to which has been added 15 g of arsenic (0.015%). LEAD 251: NMT 20 ppm SPECIFIC TESTS COMPLETENESS OF SOLUTION 641: Meets the requirements, using a 750-mg specimen and water as the solvent LOSS ON DRYING 731: Dry at 105 to constant weight: it loses NMT 2.7% of its weight. ACIDITY OR ALKALINITY Sample solution: 1.0 g in 50 mL of carbon dioxide-free water Analysis: Titrate with 0.010 N hydrochloric acid or 0.010 N sodium hydroxide to a pH of 4.5.
Cryoprecipitated Antihemophilic Factor

(Comment on this Monograph)id=m5260=Cryoprecipitated Antihemophilic Factor=A-Monos.pdf) DEFINITION Cryoprecipitated Antihemophilic Factor conforms to the regulations of the FDA concerning biologics (640.50 to 640.57) (see Biologics 1041). It is a sterile, frozen concentrate of human antihemophilic factor prepared from the Factor VIIIrich cryoprotein fraction of human venous plasma obtained from suitable whole-blood donors from a single unit of plasma derived from whole blood or by plasmapheresis, collected and processed in a closed system. It contains no preservative. It meets the requirements of the test for potency by comparison with the U.S. Standard Antihemophilic Factor (Factor VIII) or with a working reference that has been calibrated with it, in having an average potency of NLT 80 Antihemophilic Factor Units/container, made at intervals of NMT 1 month during the dating period. SPECIFIC TESTS EXPIRATION DATE: The expiration date is not later than 1 year from the date of collection of source material. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in hermetic containers at a temperature of 18 or lower. LABELING: Label it to indicate the ABO blood group designation and the identification number of the donor from whom the source material was obtained. Label it also with the type and result of a serologic test for syphilis, or to indicate that it was non-reactive in such test; with the type and result of a test for hepatitis B surface antigen, or to indicate that it was non-reactive in such test; with a warning not to use it if there is evidence of breakage or thawing; with instructions to thaw it before use to a temperature between 20 and 37, after which it is to be stored at room temperature and used as soon as possible but within 6 h after thawing; to state that it is to be used within 4 h after the container is entered; and to state that it is for intravenous administration, and that a filter is to be used in the administration equipment.
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Antimony / Official Monographs

NMT 2.0 mL is required.
USP 32
Analysis: Titrate with 0.010 N hydrochloric acid or 0.010 N sodium hydroxide to a pH of 4.5 Acceptance criteria: NMT 2.0 mL is required. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Antimony Sodium Tartrate

(Comment on this Monograph)id=m5300=Antimony Sodium Tartrate=A-Monos.pdf)
Antipyrine
(Comment on this Monograph)id=m5350=Antipyrine=AMonos.pdf)
C8H4Na2O12Sb2 Antimonate(2-), bis[-[2,3-dihydroxybutanedioato(4-)O1,O2:O3,O4]]di-, disodium, stereoisomer; Disodium bis[-[L-(+)-tartrato(4-)]]diantimonate(2-) [34521-09-0].
581.61 C11H12N2O 1,2-Dihydro-1,5-dimethyl-2-phenyl-3H-pyrazol-3-one; 2,3-Dimethyl-1-phenyl-3-pyrazolin-5-one [60-80-0]. DEFINITION Antipyrine contains NLT 99.0% and NMT 100.5% of C11H12N2O, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Wavelength: 266 nm Sample solution: 20 g/mL in methanol Acceptance criteria: Absorptivities calculated on the dried basis, do not differ by more than 3.0% C. PROCEDURE Analysis: Add tannic acid TS to a solution of it. Acceptance criteria: A white precipitate is formed. ASSAY PROCEDURE Sample: 150 mg of Antipyrine Analysis: Dissolve Sample in 25 mL of water in a 250-mL iodine flask. Add 2 g of sodium acetate, 1 mL of diluted acetic acid, and 20.0 mL of 0.1 N iodine VS and allow to stand in a cool, dark place for 20 min. Add 25 mL of alcohol to dissolve the precipitate, and titrate the excess iodine with 0.1 N sodium thiosulfate VS, using starch TS as the indicator. Each mL of 0.1 N iodine is equivalent to 9.412 mg of C11H12N2O. 188.23
DEFINITION Antimony Sodium Tartrate contains NLT 98.0% and NMT 101.0% of C8H4Na2O12Sb2, calculated on the dried basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Antimony 191, Sodium 191, and Tartrate 191 ASSAY PROCEDURE Sample: 500 mg of Antimony Potassium Tartrate Analysis: Dissolve in 50 mL of water, add 5 g of potassium sodium tartrate, 2 g of sodium borate, and 3 mL of starch TS, and immediately titrate with 0.1 N iodine VS to the production of a persistent blue color. Each mL of 0.1 N iodine is equivalent to 14.54 mg of C8H4Na2O12Sb2. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities ARSENIC, Method II 211: NMT 8 ppm LEAD 251: NMT 20 ppm SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 105 to constant weight: loses NMT 6.0% of its weight. ACIDITY OR ALKALINITY Sample solution: 1.0 g in 50 mL of carbon dioxide-free water
USP 32
Official Monographs / Antipyrine 261

Acceptance criteria: The IR absorption spectrum of this solution exhibits maxima at the same wavelengths as that of a similar solution of USP Antipyrine RS, concomitantly measured. C. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 7.7 mg/mL of ammonium acetate in water Mobile phase: Acetonitrile and Solution A (1:3) Standard solution: Transfer 15 mg of USP Benzocaine RS to a 100-mL volumetric flask. Add 15 J mg of USP Antipyrine RS, accurately weighed, J being the ratio of the labeled amount, in mg, of antipyrine to the labeled amount, in mg, of benzocaine per mL of Otic Solution. Dissolve in 50 mL of methanol, and dilute with methanol to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, dilute with Mobile phase to volume, and mix. Sample solution: Transfer an accurately measured volume of Otic Solution, equivalent to about 15 mg of benzocaine, to a 100-mL volumetric flask. Dissolve in 50 mL of methanol, dilute with methanol to volume, and mix. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, dilute with Mobile phase to volume, and mix Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4-mm 15-cm; 5-m packing L15 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for antipyrine and benzocaine are 0.35 and 1.0, respectively.] Suitability requirements Column efficiency: NLT 1500 theoretical plates, benzocaine peak Tailing factor: NMT 2.5, benzocaine peak Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H11NO2 in each mL of the Otic Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response due to benzocaine in the Sample solution = peak response due to benzocaine in the rS Standard solution = concentration of USP Benzocaine RS in the CS Standard solution (mg/mL) = nominal concentration of benzocaine in the CU Sample solution (mg/mL) Calculate the percentage of C11H12N2O in each mL of the Otic Solution taken by the same formula, changing the terms to refer to antipyrine instead of benzocaine. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1.0% rU
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.15% HEAVY METALS, Method II 231: NMT 20 ppm Sample solution: 1 g in 2 mL of 1 N acetic acid, and add water to make 25 mL Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Standard solution: Chloroform Sample solution: Chloroform Eluant: Chloroform, acetone, butyl alcohol, and formic acid (60:15:15:15) Visualization: 1 SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 110112.5 LOSS ON DRYING 731: Dry it at 60 for 2 h: it loses NMT 1.0% of its weight. COMPLETENESS AND COLOR OF SOLUTION: It is completely soluble in its own weight of cold water, the solution being colorless or not more than slightly yellow when viewed transversely in a tube having a diameter of 20 mm. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers USP REFERENCE STANDARDS 11 USP Antipyrine RS
Antipyrine and Benzocaine Otic Solution

(Comment on this Monograph)id=m5360=Antipyrine and Benzocaine Otic Solution=A-Monos.pdf) DEFINITION Antipyrine and Benzocaine Otic Solution is a solution of Antipyrine and Benzocaine in Glycerin. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of antipyrine (C11H12N2O) and benzocaine (C9H11NO2). [NOTEIn the preparation of this Otic Solution, use Glycerin that has a low water content, in order that the Otic Solution may comply with the limit for Water determination. This may be ensured by using Glycerin having a specific gravity of NLT 1.2607, corresponding to a concentration of 99.5%.] IDENTIFICATION A. PROCEDURE Sample: 5 mL Analysis: Transfer the Sample to a separator containing 25 mL of water, and extract the solution with two 25-mL portions of a mixture of equal volumes of ether and solvent hexane. Combine the extracts, and retain the water solution for Identification test B. Extract the etherhexane solution with 50 mL of water, and discard the water layer. Evaporate the etherhexane solution to dryness, dry the residue in a vacuum at 40 to 50 for 1 h, and dissolve the residue in 1 mL of chloroform. Acceptance criteria: The IR absorption spectrum of this solution exhibits maxima at the same wavelengths as that of a similar solution of USP Benzocaine RS, concomitantly measured. B. PROCEDURE Analysis: Add 5 mL of 1 N sodium hydroxide solution to the water solution retained from Identification test A, and extract with two 25-mL portions of chloroform. Evaporate the combined extracts to dryness, dry the residue in a vacuum at 4050 for 1 h, and dissolve the residue in 3 mL of chloroform.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Antipyrine RS USP Benzocaine RS
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Antipyrine / Official Monographs

USP 32
Antipyrine, Benzocaine, and Phenylephrine Hydrochloride Otic Solution

(Comment on this Monograph)id=m5365=Antipyrine, Benzocaine, and Phenylephrine Hydrochloride Otic Solution=AMonos.pdf) DEFINITION Antipyrine, Benzocaine, and Phenylephrine Hydrochloride Otic Solution is a solution of Antipyrine, Benzocaine, and Phenylephrine Hydrochloride in a suitable nonaqueous solvent. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of antipyrine (C11H12N2O), benzocaine (C9H11NO2), and phenylephrine hydrochloride (C9H13NO2 HCl). IDENTIFICATION The retention times of the major peaks of the Sample solutions correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, 0.005 M solution of sodium 1heptanesulfonate in water, and phosphoric acid (120:880:1) Standard solution: 25 mg of USP Antipyrine RS, 25 mg of USP Benzocaine RS, and 25 mg of USP Phenylephrine Hydrochloride RS in a 250-mL volumetric flask. Add 5 mL of a 0.5 mg/mL solution of p-aminobenzoic acid in Mobile phase. Add 150 mL of Mobile phase, and mix to dissolve, sonicating if necessary. Dilute with Mobile phase to volume. Sample solution A: Nominally equivalent to 0.1 mg/mL of antipyrine, from Otic Solution diluted in Mobile phase Sample solution B: Nominally equivalent to 0.1 mg/mL of benzocaine, from Otic Solution diluted in Mobile phase Sample solution C: Nominally equivalent to 0.1 mg/mL of phenylephrine hydrochloride, from Otic Solution diluted in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 272 nm Column: 4.6-mm 30-cm; packing L11 Flow rate: 1.5 mL/min Injection size: 20 or 25 L System suitability Sample: Standard solution [NOTEThe relative retention times of p-aminobenzoic acid, phenylephrine, antipyrine, and benzocaine are about 0.19, 0.26, 0.64, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between phenylephrine and aminobenzoic acid Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solutions Calculate the percentage of C11H12N2O in each mL of the Otic Solution taken: Result = (rU/rS) (CS/CU) 100 peak response from Sample solution A peak response from the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of antipyrine in the Sample solution (mg/mL) Calculate the percentages of C9H11NO2 and C9H13NO2 HCl in each mL of the Otic Solution taken, changing the terms to refer to benzocaine or phenylephrine hydrochloride instead of antipyrine. rU rS CS CU = = = =
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Antipyrine RS USP Benzocaine RS USP Phenylephrine Hydrochloride RS
Antithrombin III Human

(Comment on this Monograph)id=m5390=Antithrombin III Human=A-Monos.pdf) DEFINITION Antithrombin III Human is a glycoprotein, which is the major inhibitor of thrombin and other activated clotting factors, including factors IX, X, XI, and XII, and the cofactor through which heparin exerts its effect. It is obtained from human plasma of healthy donors who must, as far as can be ascertained, be free from detectable agents of infection transmissible by transfusion of blood or blood derivatives. The method of manufacturing includes steps that have been shown to remove or inactivate known agents of infection. If substances are used for inactivation of viruses during production, the subsequent purification procedure must be validated to demonstrate that the concentration of these substances is reduced to an acceptable level and any residues are such as not to compromise the safety of the preparation for patients. The antithrombin III concentrate is passed through a bacteria-retentive filter, filled aseptically into its final, sterile containers, and immediately frozen. It is then freezedried, and the containers are closed under vacuum. No antimicrobial preservative is added at any stage of production. Antithrombin III Human complies with the requirements for Biologics 1041. When reconstituted in the recommended volume of diluent, the potency is NLT 25 USP Antithrombin III Units/mL. [NOTEOne USP Antithrombin III Unit is the amount of antithrombin III that forms a complex with one unit of thrombin at 25 in the presence of heparin at a pH of 8.4.] IDENTIFICATION Meets the requirements of the Assay. ASSAY PROCEDURE Solution A: Mix Tris(hydroxymethyl)aminomethane, edetic acid, and sodium chloride in water containing 0.1% polyethylene glycol 6000 to obtain a solution having concentrations of 0.050 M, 0.0075 M, and 0.175 M, respectively. Adjust with hydrochloric acid or sodium hydroxide solution to a pH of 8.4. Solution B: 0.05% (w/v) of albumin human in Solution A Solution C: 10 mg/mL of polybrene in Solution B Solution D: 15 USP Heparin Units/mL of USP Heparin Sodium RS in Solution B Solution E: Reconstitute thrombin bovine, and dilute with Solution B to obtain a solution having a concentration of 2.0 Thrombin Units/mL. Solution F: Prepare a solution of chromogenic substrate for amidolytic test (see Reagents, Indicators, and Solutions Reagent Specifications) for factor IIa in water to obtain a solution having a concentration of about 5.0 mM, and dilute the solution further with Solution C to 1.0 mM.
USP 32
Stopping solution: 20% of acetic acid Standard solution A: USP Antithrombin III Human RS in Solution D to obtain a solution containing 1.0 USP Antithrombin III Unit Standard solutions B, C, D, and E: Dilute Standard solution A with Solution D 60-, 120-, 180-, and 300-fold. Sample solution A: Dissolve a quantity of Antithrombin III Human in Solution D to obtain a solution having the same concentration as Standard solution A. Sample solutions B, C, D, and E: Dilute Sample solution A with Solution D 60-, 120-, 180-, and 300-fold. Analysis: Pipet 400 L each of Standard solutions B, C, D, and E and Sample solutions B, C, D, and E into suitable tubes placed in a water bath set at 37. Add 200 L of Solution E, prewarmed at 37 to each tube, mix, and incubate for 1 min. Add 200 L of Solution F prewarmed at 37 to each tube, mix, and incubate for 60 s. Stop the reaction by adding 200 L Stopping solution. To prepare a blank, add the reagents in reverse order, starting with 200 L of Stopping solution, followed by the addition of 200 L of Solution F, then adding 200 L of Solution E, and ending with 400 L of Solution D. Record the absorbance at 405 nm against the blank. For Standard solutions and Sample solutions, calculate the regression of the absorbance against log concentrations, and calculate the activity of Antithrombin III Human in USP Antithrombin III Units, using a suitable statistical method for parallel-line assays. The four independent relative activity estimates are then combined to obtain the final mean, and the confidence limits are calculated. Acceptance criteria: 80%120%. The specific activity is NLT 6.0 USP Antithrombin III Units/mg of total protein. The confidence interval (P = 0.95) is between 90% and 110%. IMPURITIES Organic Impurities PROCEDURE: HEPARIN CONTENT Solution A: Mix Tris(hydroxymethyl)aminomethane, edetic acid, and sodium chloride in water containing 0.1% polyethylene glycol 6000 to obtain a solution having concentrations of 0.050 M, 0.0075 M, and 0.175 M, respectively. Adjust with hydrochloric acid or sodium hydroxide solution to a pH of 8.4. Solution B: Solution of chromogenic substrate for amidolytic test for factor Xa in water to obtain a solution of concentration of 2.5 mM Solution C: Factor Xa in Solution A to obtain a solution containing 20 nanokatalytic units (nkats) Solution D: 20% (v/v) of acetic acid in water Standard solution: USP Antithrombin III Human RS in Solution A to obtain a solution containing 1.0 USP Antithrombin III Unit Sample solution: Antithrombin III Human in Solution A to obtain a solution containing 1.0 USP Antithrombin III Unit Analysis: Pipet 250 L each of Solution A, the Standard solution, and the Sample solution to suitable tubes placed in a water bath set at 37. Add 250 L of Solution C prewarmed at 37 to each tube, and incubate for 2 min. Add 250 L of Solution B prewarmed at 37 to each tube, mix, and incubate for 120 s. Stop the reaction by adding 250 L of Solution D. Record the absorbance at 405 nm, using Solution A as the blank. Calculate the USP Heparin Unit/USP Antithrombin III Unit: Result = PR (AF AU)/(AF AS) PR AF AU AS = heparin content of USP Antithrombin III Human RS in USP Heparin Unit/USP Antithrombin III Unit = absorbance values from Solution A = absorbance values from the Sample solution = Absorbance values from the Standard solution
Official Monographs / Antithrombin 263

Acceptance criteria: NMT 0.1 USP Heparin Unit/USP Antithrombin III Unit. SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Direct Inoculation of the Culture Medium WATER DETERMINATION, Method I 921: NMT 3.0% PYROGEN TEST 151: Inject 50 USP Antithrombin III Units/kg of the rabbits weight, calculated from the activity stated on the label. Meets the requirements. GENERAL SAFETY: Meets the requirements for biologics as set forth for under Biological Reactivity Tests, In Vivo 88, Safety TestsBiologicals OSMOLALITY AND OSMOLARITY 785: Reconstitute with the diluent according to the manufacturers instruction: NLT 240 mOsmol/kg for the solution. PH 791: Reconstitute with the diluent according to the manufacturers instruction: 6.07.5. MOLECULAR WEIGHT DISTRIBUTION Mobile phase: Solution containing 0.1 M sodium phosphate, 0.15 M sodium chloride, and 0.05% sodium azide, having a pH of 6.5 Solution A: 45 mg/mL of thyroglobulin in Mobile phase Sample solution: 810 mg/mL of Antithrombin III Human System suitability solution: Dilute USP Albumin Human RS, if necessary, with water to obtain a solution containing 5%. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 7.5- 75-mm guard column and a 7.5- 300mm analytical column, both containing packing L59 Temperature: Ambient Flow rate: 0.5 mL/min maintained constant to 1% Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Column efficiency: Greater than 1500 theoretical plates Tailing factor: 0.52.5 Analysis Samples: Solution A and Sample solution Acceptance criteria: Note the retention times of the major peak in the Solution A chromatogram. The relative peak area of the high molecular weight peak eluting at about the same retention time as the major peak in the Solution A chromatogram, or earlier, is NMT 13%. TOTAL PROTEIN CONTENT Solution A: 1000 mg/mL of trichloroacetic acid Sample solution: 7.5 mg/mL of Antithrombin III Human in 0.15 M sodium chloride solution Blank: 0.15 M solution of sodium chloride Analysis: To each of 2.0 mL of the Sample solution and the Blank in suitable centrifuge tubes, add 1.5 mL of Solution A. Mix, allow to stand for at least 10 min, centrifuge for 5 min, and decant the supernatant. Resuspend the precipitates in 1.5 mL of Solution A, centrifuge for 5 min, decant the supernatant, and hold the tubes inverted on a filter paper to drain. Transfer the residues with a minimum quantity of water to a micro-Kjeldahl flask, and determine the nitrogen content using Method II (see Nitrogen Determination 461). Multiply the result, corrected for the Blank, by 6.25 to calculate the quantity of protein. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Use a Type I glass container with an appropriate stopper and seal. Store protected from light between 2 and 8, excursions permitted up to 25. LABELING: The labeling should state the content of antithrombin III in USP Antithrombin III Units. The diluent
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USP 32
LABELING: Label it to indicate the species of spider against which the Antivenin is to be used, that it is not intended to protect against bites from other spider species, and to state that it was prepared in the horse. EXPIRATION DATE: The expiration date for Antivenin containing a 10% excess of potency is NMT 5 years after the date of issue from manufacturers cold storage (5, 1 year; 0, 2 years).
and the volume to be used to reconstitute the preparation are indicated. USP REFERENCE STANDARDS 11 USP Albumin Human RS USP Antithrombin III Human RS USP Heparin Sodium RS
Antivenin (Crotalidae) Polyvalent

(Comment on this Monograph)id=m5410=Antivenin (Crotalidae) Polyvalent=A-Monos.pdf) DEFINITION Antivenin (Crotalidae) Polyvalent conforms to the regulations of the FDA concerning biologics (see Biologics 1041). It is a sterile, non-pyrogenic preparation derived by drying a frozen solution of specific venom-neutralizing globulins obtained from the serum of healthy horses immunized against the venoms of four species of pit vipers, Crotalus atrox, Crotalus adamanteus, Crotalus durissus terrificus, and Bothrops atrox (Fam. Crotalidae). It is standardized by biological assay on mice, in terms of one dose of antivenin neutralizing the venoms in NLT the number of mouse LD50 stated, of Crotalus atrox (Western diamondback), 180; Crotalus durissus terrificus (South American rattlesnake), 1320; and Bothrops atrox (South American fer-delance), 780. It may contain a suitable preservative. When constituted as specified in the labeling, it is opalescent and contains NMT 20.0% of solids, determined by drying 1 mL at 105 to constant weight (1 mg). SPECIFIC TESTS SAFETY: It meets the requirements for general safety (see Biological Reactivity Tests, In Vivo 88, Safety Tests Biologicals). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers. Avoid exposure to excessive heat. LABELING: Label it to indicate the species of snakes against which the Antivenin is to be used, and to state that it was prepared from horse serum. EXPIRATION DATE: The expiration date for Antivenin containing a 10% excess of potency is NMT 5 years after the date of issue from manufacturers cold storage (5, 1 year; or 0, 2 years).
Antivenin (Micrurus fulvius)

(Comment on this Monograph)id=m5440=Antivenin (Micrurus fulvius)=A-Monos.pdf) DEFINITION Antivenin (Micrurus fulvius) conforms to the regulations of the FDA concerning biologics (see Biologics 1041). It is the sterile, non-pyrogenic preparation derived by drying a frozen solution of specific venom-neutralizing globulins obtained from the serum of healthy horses immunized against venom of the Eastern Coral snake (Micrurus fulvius). It is standardized by biological assay on mice, in terms of one dose of antivenin neutralizing the venom of Micrurus fulvius in NLT 250 mouse LD50. It may contain a suitable preservative. When constituted as specified in the labeling, it is opalescent and contains NMT 20.0% of solids, determined by drying 1 mL at 105 to constant weight ( 1 mg). SPECIFIC TESTS SAFETY: It meets the requirements for general safety (see Biological Reactivity Tests, In Vivo 88, Safety Tests Biologicals). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, and avoid exposure to excessive heat. LABELING: Label it to indicate the species of snake against which the Antivenin is to be used, and to state that it was prepared in the horse. EXPIRATION DATE: The expiration date for Antivenin containing a 10% excess of potency is NMT 5 years after the date of issue from the manufacturers cold storage (5, 1 year; or 0, 2 years).
Apomorphine Hydrochloride Antivenin (Latrodectus mactans)

(Comment on this Monograph)id=m5420=Antivenin (Latrodectus mactans)=A-Monos.pdf) DEFINITION Antivenin (Latrodectus mactans) conforms to the regulations of the FDA concerning biologics (see Biologics 1041). It is the sterile, non-pyrogenic preparation derived by drying a frozen solution of specific venom-neutralizing globulins obtained from the serum of healthy horses immunized against the venom of black widow spiders (Latrodectus mactans). It is standardized by biological assay on mice, in terms of one dose of antivenin neutralizing the venom of Latrodectus mactans in NLT 6000 mouse LD50. Thimerosal 1:10,000 is added as a preservative. When constituted as specified in the labeling, it is opalescent and contains NMT 20.0% of solids. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers. Avoid exposure to excessive heat. (Comment on this Monograph)id=m5660=Apomorphine Hydrochloride=A-Monos.pdf)
312.79 C17H17NO2 HCl 1/2H2O 303.79 C17H17NO2 HCl 4H-Dibenzo[de,g]quinoline-10,11-diol, 5,6,6a,7-tetrahydro-6methyl-, hydrochloride, hemihydrate, (R)-; 6a-Aporphine-10,11-diol hydrochloride hemihydrate [41372-20-7]. Anhydrous [314-19-2]. DEFINITION Apomorphine Hydrochloride contains NLT 98.5% and NMT 100.5% of C17H17NO2 HCl, calculated on the dried basis.
USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Analysis: To 5 mL of a solution (1 in 100) add a slight excess of sodium bicarbonate solution (1 in 20): a white or greenish-white precipitate is formed. Add 3 drops of iodine TS, and shake vigorously: an emerald-green color is produced. Add 5 mL of ether and, after vigorous shaking, allow the layers to separate: the ether is colored deep rubyred while the water layer retains its green color. C. PROCEDURE Analysis: Dissolve it in nitric acid. Acceptance criteria: A dark purple solution is produced. D. PROCEDURE Analysis: To a solution of it add silver nitrate TS. Acceptance criteria: A white precipitate, which is insoluble in nitric acid, is formed. This precipitate soon darkens by reduction to metallic silver, the reduction being accelerated by the addition of 6 N ammonium hydroxide. ASSAY PROCEDURE Sample solution: Dissolve 300 mg of Apomorphine Hydrochloride in 100 mL of glacial acetic acid with the aid of heat provided by a steam bath. Add 0.1 mL of acetic anhydride to the hot solution, and stir for 5 min. Cool to room temperature, and add 5 mL of mercuric acetate TS and 0.25 mL of crystal violet TS. Analysis Sample: Sample solution Titrate with 0.1 N perchloric acid VS to a blue endpoint. Perform a blank determination and perform any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 30.38 mg of C17H17NO2 HCl. Acceptance criteria: 98.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE 1: ORDINARY IMPURITIES 466 Solvent for sample solution: Methanol Solvent for standard solution: Methanol Eluant: 1-Butanol, water, and formic acid (7:2:1) Visualization: A freshly prepared mixture of 10% ferric chloride solution and 5% potassium ferricyanide solution (2:1). Allow the chromatograms to develop until the solvent front has moved about 8 cm. [NOTEThe development time is 1.52 h.] Allow the plates to dry at room temperature for 1 h prior to spraying. PROCEDURE 2: DECOMPOSITION PRODUCTS Analysis: Shake 100 mg with 5 mL of ether. Acceptance criteria: The latter acquires NMT a pale reddish color. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 60.5 to 63.0 Sample solution: 15 mg/mL in dimethylsulfoxide LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses 2.0%3.5% of its weight. COLOR OF SOLUTION: Place 100 mg in a suitable test tube, add 10 mL of cold, oxygen-free water, and agitate gently until dissolved: the color of the resulting solution, observed promptly after the Apomorphine Hydrochloride has dissolved, is not more intense than that of a color standard prepared as follows. Dissolve 5 mg of Apomorphine Hydrochloride in 100.0 mL of water. Transfer 1.0 mL of this solution to a test tube of the same size as that used for the Sample solution, dilute with 6 mL of water, add 1 mL of
Official Monographs / Apomorphine 265

sodium bicarbonate solution (1 in 20), and then add 0.50 mL of iodine TS. Allow to stand for 30 s, add 0.60 mL of sodium thiosulfate solution (1 in 40), and dilute with water to 10 mL. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Apomorphine Hydrochloride RS
Apomorphine Hydrochloride Tablets

(Comment on this Monograph)id=m5690=Apomorphine Hydrochloride Tablets=A-Monos.pdf) DEFINITION Apomorphine Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C17H17NO2 HCl 1 /2H2O. IDENTIFICATION PROCEDURE Analysis: To 5 mL of a filtered solution of Tablets, containing about 10 mg of apomorphine hydrochloride, add a slight excess of sodium bicarbonate solution (1 in 20): a white or greenish-white precipitate is formed. Add 3 drops of iodine TS, and shake vigorously: an emerald-green color is produced. Add 5 mL of ether, and, after vigorous shaking, allow the layers to separate: the ether is colored deep rubyred while the water layer retains its green color. ASSAY PROCEDURE Sample solution: Dissolve an equivalent to 50 mg of apomorphine hydrochloride, from finely powdered tablets (NLT 20) in 25 mL of water in a separator. Add 500 mg of sodium bicarbonate, and completely extract with successive small portions of ether. Combine the ether extracts in a separator, and wash them with three 5-mL portions of water. Shake the combined water washings with 10 mL of ether, and add this ether to the combined ether extracts. Extract the ether solutions with 20.0 mL of 0.02 N sulfuric acid VS, and wash with three 5-mL portions of water. Combine the acid extract and washings in a beaker, and warm on a steam bath to expel any residual ether. Cool and add methyl red TS. Analysis: Titrate the excess acid with 0.02 N sodium hydroxide VS (see Titrimetry 541, Residual Titrations). Each mL of 0.02 N sulfuric acid is equivalent to 6.256 mg of C17H17NO2 HCl 1/2H2O. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701 Time: 15 min UNIFORMITY OF DOSAGE UNITS 905 Procedure for content uniformity Sample solution: Place 1 Tablet in a 500-mL volumetric flask containing 100 mL of 0.1 N hydrochloric acid, and shake for 15 min. Dilute with 0.1 N hydrochloric acid to volume, mix, and filter, discarding the first 20 mL of filtrate. Dilute a portion of the subsequent filtrate quantitatively and stepwise, if necessary, with 0.1 N hydrochloric acid to provide a solution containing approximately 12 g/mL of apomorphine hydrochloride. Standard solution: 12 g/mL of anhydrous apomorphine hydrochloride, from USP Apomorphine Hydrochloride RS in the same medium
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USP 32
DEFINITION Apraclonidine Hydrochloride contains NLT 98.0% and NMT 102.0% of C9H10Cl2N4 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Chloride 191: requirements
Spectrometric conditions Analytical wavelength: 273 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis Samples: Sample solution and Standard solution Concomitantly determine the absorbances of the Sample solution and the Standard solution. Calculate the percentage of C17H17NO2 HCl 1/2H2O in the Tablet taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of anhydrous apomorphine hydrochloride in the Standard solution (g/mL) CU = concentration of apomorphine hydrochloride in the solution from the Tablet, based upon the labeled quantity per Tablet and the extent of dilution (g/mL) = molecular weight of apomorphine hydrochloride Mr1 hemihydrate, 312.80 = molecular weight of anhydrous apomorphine Mr2 hydrochloride, 303.79 Acceptance criteria: Meet the requirements SPECIFIC TESTS COLOR OF SOLUTION: Dissolve a quantity of powdered Tablets, equivalent to 5 mg of apomorphine hydrochloride, in water to make 100.0 mL. Transfer 1.0 mL of the solution to a test tube, dilute with 6 mL of water, and, if necessary, filter through a small pledget of cotton. Add 1 mL of sodium bicarbonate solution (1 in 20), then add 0.50 mL of iodine TS. Allow to stand for 30 s, then add 0.60 mL of sodium thiosulfate solution (1 in 40), and dilute with water to 10 mL. This solution represents the color standard. Place a quantity of powdered Tablets, equivalent to about 50 mg of apomorphine hydrochloride, in a test tube of suitably small size. Add 10.0 mL of cold, oxygen-free water, insert the stopper in the test tube, and agitate gently until no more dissolves. If necessary, filter immediately through a small pledget of cotton. The color of the solution, observed promptly after preparation, is not more intense than that of the color standard. Use closely matched test tubes for the comparison. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Apomorphine Hydrochloride RS AU AS CS
Meets the
ASSAY PROCEDURE Sample solution: 125 mg of Apraclonidine Hydrochloride in 40 mL of glacial acetic acid. Add 10 mL of mercuric acetate TS. Analysis: Titrate with 0.1 N perchloric acid VS, using a calomel-glass electrode system (see Titrimetry 541). Perform a blank determination. Each mL of 0.1 N perchloric acid is equivalent to 14.08 mg of C9H10Cl2N4 HCl. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Phosphate buffer: 6.8 mL of phosphoric acid in a 2000mL volumetric flask. Add 1900 mL of water. Adjust with sodium hydroxide solution (1 in 2) to a pH of 3.0, and dilute with water to volume. Mobile phase: Acetonitrile, methanol, and Phosphate buffer (14:1:10) System suitability solution: 0.8 mg/mL of USP Apraclonidine Hydrochloride RS in Mobile phase Sample solution: 0.8 mg/mL of Apraclonidine Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 8-mm 100-mm; packing L7 Flow rate: 3 mL/min Injection size: 20 L System suitability Sample: System suitability solution Suitability requirements Tailing factor: NMT 2.2 for the apraclonidine peak Relative standard deviation: NMT 2.0% Analysis Sample: Sample solution [NOTEAllow about five times the elution time of apraclonidine before making the next injection.] Calculate the percentage of each peak, other than the solvent peak and the apraclonidine peak, in the portion of Apraclonidine Hydrochloride taken: Result = (rU/rT) 100 = response of each peak other than the principal peak = sum of the responses of all of the peaks, rT excluding that of the solvent peak Acceptance criteria Individual impurities: NMT 1.0% Total impurities: NMT 2.0% rU SPECIFIC TESTS PH 791: 5.06.6, in a solution (1 in 100) LOSS ON DRYING 731: Dry it in a vacuum at 105 for 3 h: it loses NMT 1.0% of its weight.
Apraclonidine Hydrochloride
(Comment on this Monograph)id=m5700=Apraclonidine Hydrochloride=A-Monos.pdf)
281.57 C9H10Cl2N4 HCl 1,4-Benzenediamine, 2,6-dichloro-N1-2-imidazolidinylidene-, monohydrochloride; 2-[(4-Amino-2,6-dichlorophenyl)imino]imidazolidine monohydrochloride [73218-79-8].
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Apraclonidine Hydrochloride RS
Official Monographs / Apraclonidine 267

Standard solution: 11.5 g/mL of USP Apraclonidine Hydrochloride RS (equivalent to about 10 g of apraclonidine per mL) diluted in Mobile phase System suitability solution: Transfer 1 mL of propiophenone to a 100-mL volumetric flask, and dilute with methanol to volume. Transfer 3.0 mL of this solution to a 50-mL volumetric flask, and dilute with methanol to volume. Transfer 1.0 mL of this solution and 5.0 mL of the Standard stock solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Sample stock solution: Equivalent to 0.2 mg/mL of apraclonidine, from Ophthalmic Solution in water Sample solution: 10 g/mL of Sample stock solution diluted in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 8-mm 100-mm; packing L7 Flow rate: 3 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for apraclonidine and propiophenone are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the analyte and propiophenone peaks Column efficiency: NLT 1000 theoretical plates for the analyte peak Tailing factor: NMT 2.2 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H10Cl2N4 in the portion of Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) x 100 = peak response of apraclonidine from the Sample solution = peak response of apraclonidine from the rS Standard solution = concentration of USP Apraclonidine CS Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of apraclonidine in the CU Sample solution (g/mL) = molecular weight of apraclonidine, 245.11 Mr1 = molecular weight of apraclonidine hydrochloride, Mr2 281.57 Acceptance criteria: 90.0%115.0% rU SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 4.47.8 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Apraclonidine Hydrochloride RS
Apraclonidine Ophthalmic Solution

(Comment on this Monograph)id=m5705=Apraclonidine Ophthalmic Solution=A-Monos.pdf) DEFINITION Apraclonidine Ophthalmic Solution is a sterile, aqueous solution of Apraclonidine Hydrochloride. It contains an amount of apraclonidine hydrochloride (C9H10Cl2N4 HCl) equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of apraclonidine (C9H10Cl2N4). IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the major peak of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST (see Chromatography 621.) Adsorbent: 0.2-mm layer of chromatographic silica gel mixture, or equivalent Standard solution: 11.5 mg/mL of USP Apraclonidine Hydrochloride RS in methanol Sample solution: Apraclonidine Ophthalmic Solution Application volume: 2 L Developing solvent system: Chloroform, methanol, and ammonium hydroxide (37:11:2) Spray reagent: 1 mg/mL of fluorescamine in acetone Analysis Samples: Standard solution and Sample solution Allow the applications to dry, and develop the chromatogram in the developing solvent system, until the solvent front has moved about three-fourths of the length of the plate. Locate the spots on the plate by viewing under short-wavelength UV light. [NOTEThe apraclonidine spot should appear as a blue spot.] Spray the plate with Spray reagent. [NOTEAvoid prolonged or repeated breathing of the aerosol from the fluorescamine spray. Also avoid prolonged or repeated contact with skin. Fluorescamine solution should be sprayed only in a hood.] Examine the plate under normal light and longwavelength UV light. [NOTEThe apraclonidine spot should appear as a yellow spot under normal light and as a white spot under long-wavelength UV light.] Acceptance criteria: The RF value and appearance of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Phosphate buffer: 6.8 mL of phosphoric acid in a 2000-mL volumetric flask, and add 1900 mL of water. Adjust with sodium hydroxide solution (1 in 2) to a pH of 3.0, and dilute with water to volume. Mobile phase: Acetonitrile, methanol, and Phosphate buffer (15:1:34) Standard stock solution: 0.23 mg/mL of USP Apraclonidine Hydrochloride RS in water
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immediately with 0.0015 M borate buffer to 40.0 mL. Allow to stand at room temperature for 10 min, and then keep in ice water. [NOTEUse within 6 h of preparation.] Dilute trypsin solution: Trypsin solution and 0.0015 M borate buffer (0.5:9.5) Allow to stand at room temperature for 10 min, and then keep in ice water. Substrate solution: 6.9 mg/mL of N-benzoyl-L-arginine ethyl ester hydrochloride [NOTEUse within 2 h.] Analysis: Mix 9.0 mL of 0.0015 M borate buffer and 1.0 mL of Substrate solution in a jacketed glass vessel with a capacity of 30 mL and that contains a stirring device. The lid of the reaction vessel should contain five holes to accommodate the electrodes, the tip of a buret, a tube for the admission of nitrogen, and the introduction of reactants. An automated or manual titration apparatus may be used. Adjust to a pH of 8.0 by the addition of 0.1 N sodium hydroxide VS. Maintain an atmosphere of nitrogen within the vessel, and stir continuously. When the temperature has reached equilibrium at 25 0.1, add 1.0 mL of Trypsin and aprotinin solution, and start a timer. Maintain at a pH of 8.0 by the addition of 0.1 N sodium hydroxide VS, and note the volume added every 30 s. Continue the reaction for 6 min. Determine the volume of 0.1 N sodium hydroxide added per s, in mL (n1). Carry out a similar titration using 1.0 mL of the Dilute trypsin solution. Determine the volume of 0.1 N sodium hydroxide added per s, in mL (n2). For the lyophilized powder, calculate the aprotinin activity in USP Aprotinin Units/mg: Result = 4000 (2n2 n1)/m m = quantity of Aprotinin used to prepare 1 mL of the Sample solution (mg) For the concentrated solution, calculate the USP Aprotinin Units/mL using the following formula: Result = 4000 (2n2 n1) D = dilution factor of the concentrated solution used to prepare the Sample solution Acceptance criteria: NLT 3 USP Aprotinin Units/mg potency IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF DES-ALA-APROTININ AND DES-ALA-DESGLY-APROTININ Sample solution: 47 USP Aprotinin Units/mL of Aprotinin [NOTEDilute a concentrated solution of Aprotinin with water, or weigh out Aprotinin and dissolve in water.] Standard solution: Dilute USP Aprotinin RS with water to obtain a solution having a concentration similar to that of the Sample solution. Capillary zone electrophoresis buffer: 8.21 g of monobasic potassium phosphate in 400 mL of water. Adjust with phosphoric acid to a pH of 3.0, and dilute with water to 500 ml. Capillary zone electrophoresis system (See Biotechnology-Derived ArticlesCapillary Electrophoresis 1053.) The capillary electropherograph is equipped with a 214-nm detector and a 45- to 60-cm uncoated fused silica capillary with an internal diameter of 75 m with the temperature controlled at 25. Apply a field strength of 0.2 kV/cm for 30 min, using Capillary zone electrophoresis buffer as the electrolyte in both buffer reservoirs. Electropherograph the Standard solution, and record the peak responses as directed for Analysis: the relative migration times are 0.98 for des-Ala-des-Gly-aprotinin, 0.99 for des-Ala-aprotinin, and 1 for Aprotinin. The resolution, RS, between the des-Ala-des-Gly-aprotinin and D
Aprotinin
(Comment on this Monograph)id=m5740=Aprotinin=AMonos.pdf)
6511.44 C284H432N84O79S7 Trypsin inhibitor, pancreatic basic; L-Arginyl-L-prolyl-L-aspartyl-L-phenylalanyl-L-cysteinyl-L-leucyl-Lglutamyl-L-prolyl-L-prolyl-L-tyrosyl-L-threonylglycyl-L-prolyl-Lcysteinyl-L-lysyl-L-alanyl-L-arginyl-L-isoleucyl-L-isoleucyl-Larginyl-L-tyrosyl-L-phenylalanyl-L-tyrosyl-L-asparaginyl-L-alanylL-lysyl-L-alanylglycyl-L-leucyl-L-cysteinyl-L-glutaminyl-L-threonylL-phenylalanyl-L-valyl-L-tyrosylglycylglycyl-L-cysteinyl-L-arginylL-alanyl-L-lysyl-L-arginyl-L-asparaginyl-L-asparaginyl-Lphenylalanyl-L-lysyl-L-seryl-L-alanyl-L-glutamyl-L-aspartyl-Lcysteinyl-L-methionyl-L-arginyl-L-threonyl-Lcysteinylglycylglycyl-L-alanine cyclic (555), (1438), (3051) tris(disulfide) [9087-70-1]. DEFINITION Aprotinin is a polypeptide consisting of a chain of 58 amino acid residues, which inhibits stoichiometrically the activity of several proteolytic enzymes such as chymotrypsin, kallikrein, plasmin, and trypsin. Aprotinin is obtained from bovine tissues and purified by a suitable process, and is stored as a bulk solution or lyophilized powder. Its potency, calculated on the dried basis, is NLT 3 USP Aprotinin Units/mg. In addition, the method of manufacture is validated to result in NMT 0.2 g of histamine per 3 USP Aprotinin Units using validated methods. The origin and sourcing of bovine material must be specified in compliance with FDA requirements. The manufacturing process is validated to demonstrate the clearance of potential infectious agents (i.e., viruses, TSE agents). One USP Aprotinin Unit is equivalent to 1800 Kallikrein Inhibition Units (K.I.U.). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 15 USP Aprotinin Units/mL of Aprotinin Developing solvent system: A mixture of glacial acetic acid and water (5:4) containing 100 g/L of sodium acetate Cupric chloride solution: 10 mg/mL of cupric chloride Spray reagent: 0.1 g of ninhydrin in a mixture of Cupric chloride solution, glacial acetic acid, and alcohol (6:21:70) Analysis: Proceed as directed in the chapter, except to spray the plate with the Spray reagent, and heat at 60 to visualize the spots. B. The retention time of the major peak of the Sample solution corresponds to that of the System suitability solution, as obtained in the test for Limit of N-pyroglutamyl-aprotinin and related compounds. ASSAY PROCEDURE 0.0015 M borate buffer: Transfer 0.93 g of boric acid into a 1000-mL volumetric flask, dissolve in 900 mL of water, adjust with 5 N sodium hydroxide to a pH of 8.0, and dilute with water to volume. Transfer 100 mL of this solution into a 1000-mL volumetric flask, and dilute with water to volume. Sample solution: 1.67 USP Aprotinin Units/mL (about 0.6 mg/mL) of Aprotinin in 0.0015 M borate buffer Trypsin solution: 4300 USP Trypsin Units/mL of USP Trypsin Crystallized RS in 0.001 N hydrochloric acid [NOTE Use a freshly prepared solution, and keep in ice water.] Trypsin and aprotinin solution: To 4.0 mL of the Trypsin solution, add 1.0 mL of the Sample solution. Dilute
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des-Ala-aprotinin peaks is NLT 0.8, and the resolution between the des-Ala-aprotinin and aprotinin peaks is NLT 0.5. The migration time for the Aprotinin peak is 1925 min. The tailing factor, T, of the Aprotinin peak is NMT 3 (see Chromatography 621 for calculation). The baseline is stable and shows little drift. Rinse the capillary for at least 1 min with at least 10 total capillary volumes of 0.1 N sodium hydroxide, followed by at least 10 total capillary volumes of water, and by at least 20 capillary volumes of Capillary zone electrophoresis buffer between injections. Analysis: Transfer a volume of the Sample solution, approximately 15 mL, into the anodic end of the capillary (apply differential pressure of 3.5 kPa for 3 s either by vacuum or pressure), record an electropherogram, and measure the peak areas. Calculate the percentage contents of des-Ala-des-Glyaprotinin and des-Ala-aprotinin: Result = (ri/rT) 100 = peak response corresponding to des-Ala-des-Glyaprotinin or des-Ala-aprotinin = sum of the responses of des-Ala-des-GlyrT aprotinin, des-Ala-aprotinin, and aprotinin peaks Acceptance criteria des-Ala-des-Gly-aprotinin: NMT 8.0% des-Ala-aprotinin: NMT 7.5% PROCEDURE 2: LIMIT OF N-PYROGLUTAMYL-APROTININ AND RELATED COMPOUNDS Solution A: 3.52 mg/mL of monobasic potassium phosphate and 7.26 mg/mL of dibasic sodium phosphate Solution B: 3.52 mg/mL of monobasic potassium phosphate, 7.26 mg/mL of dibasic sodium phosphate, and 66.07 mg/mL of ammonium sulfate System suitability solution: 5 USP Aprotinin Units/mL of USP Aprotinin System Suitability RS in Solution A Sample solution: 5 USP Aprotinin Units/mL of Aprotinin in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 7.5-mm 7.5-cm; packing L52 Temperature: 40 (constant temperature) Flow rate: 1 mL/min ri
Time (min) 0 21 30 31 40 Solution A 92 64 0 92 92 Solution B 8 36 100 8 8
Official Monographs / Aprotinin 269

= response of each impurity peak = sum of the responses of all peaks of the Sample solution Acceptance criteria N-pyroglutamyl-aprotinin: NMT 1.0% Any other impurity: NMT 0.5% Sum of all unknown impurities: NMT 1.0% PROCEDURE 3: LIMIT OF HIGH MOLECULAR WEIGHT PROTEINS Mobile phase: Acetonitrile, glacial acetic acid, and water (1:1:3) System suitability solution: Aprotinin solution that contains 5 USP Aprotinin Units/mL with 2% aprotinin oligomers [NOTEThis solution can be obtained by heating lyophilized aprotinin at 112 for 2 h and dissolving the solid at the specified concentration in water.] Sample solution: 5 USP Aprotinin Units/mL of Aprotinin in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: Series of three 7.8-mm 30-cm columns; packing L33 Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: System suitability solution Suitability requirements [NOTEThe relative retention times for the dimer and aprotinin are 0.9 and 1.0, respectively.] Retention time: 24.525.5 min for aprotinin Resolution: NLT 1.3 between the dimer peak and the aprotinin peak Tailing factor: NMT 2.5 for the aprotinin peak Analysis Sample: Sample solution Calculate the percentage of each oligomer peak: Result = (ri/rT) 100 = response of each peak having a retention time less than that of aprotinin monomer = sum of the responses of all peaks rT Acceptance criteria: NMT 1.0% SPECIFIC TESTS ABSORBANCE (See Spectrophotometry and Light-Scattering 851.) Prepare a solution containing 3.0 USP Aprotinin Units/mL. The solution shows an absorption maximum at 277 nm. The absorbance at the maximum is NMT 0.80. SAFETY: Prepare a solution of Aprotinin that contains 4 USP Aprotinin Units/mL using a sufficient quantity of Water for Injection. It meets the requirements when tested as directed in Biological Reactivity Tests, In Vivo 88, Safety Tests Biologicals. SPECIFIC ACTIVITY OF THE DRY RESIDUE [NOTEThis test should only be performed when product is a concentrated solution.] Analysis: Evaporate 25.0 mL of Aprotinin concentrated solution to dryness in a water bath, dry the residue at 110 for 15 h, and weigh. From the weight of the residue and the activity determined in the Assay, calculate the number of USP Aprotinin Units/mg of dry residue. Aceptance criteria: NLT 3.0 USP Aprotinin Units/mg of dried residue is found. LOSS ON DRYING 731 [NOTEThis test should only be performed on the lyophilized powder.] ri ri rT
Injection size: 40 L System suitability Sample: System suitability solution [NOTEThe relative retention times for N-pyroglutamylaprotinin and aprotinin are 0.9 and 1.0, respectively.] Suitability requirements Retention time: 1720 min for aprotinin Resolution: NLT 1.0 between N-pyroglutamyl-aprotinin and aprotinin Tailing factor: NMT 2.0 for the aprotinin peak Analysis Sample: Sample solution Calculate the percentage of each impurity peak: Result = (ri/rT) 100
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reaction vessel should contain five holes to accommodate the electrodes, the tip of a buret, a tube for the admission of nitrogen, and the introduction of reactants. An automated or manual titration apparatus may be used. Adjust to a pH of 8.0 by the addition of 0.1 N sodium hydroxide VS. Maintain an atmosphere of nitrogen within the vessel, and stir continuously. When the temperature has reached equilibrium at 25 0.1, add 1.0 mL of Trypsin and aprotinin solution, and start a timer. Maintain at a pH of 8.0 by the addition of 0.1 N sodium hydroxide VS, and note the volume added every 30 s. Continue the reaction for 6 min. Determine the volume of 0.1 N sodium hydroxide added per s, in mL (n1). Carry out a similar titration using 1.0 mL of the Dilute trypsin solution. Determine the volume of 0.1 N sodium hydroxide added per s, in mL (n2). For the lyophilized powder, calculate the aprotinin activity in USP Aprotinin Units/mg: Result = 4000 (2n2 n1)/m m = quantity of Aprotinin used to prepare 1 mL of the Sample solution (mg) For the concentrated solution, calculate the USP Aprotinin Units/mL: Result = 4000 (2n2 n1) D = dilution factor of the concentrated solution used to prepare the Sample solution Acceptance criteria: 90.0%110.0% OTHER COMPONENTS SODIUM CHLORIDE CONTENT Sample: 5.0 mL of Injection Analysis: Pipet Sample into 50 mL of water in a beaker. Add 10 mL of 25% nitric acid. Titrate with 0.1 N silver nitrate VS to a potentiometric endpoint, using a silver combination electrode. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N silver nitrate is equivalent to 5.844 mg of sodium chloride. Acceptance criteria: 42.547.5 mg IMPURITIES Organic Impurities PROCEDURE: LIMIT OF HIGH MOLECULAR WEIGHT PROTEINS Mobile phase: Acetonitrile, glacial acetic acid, and water (1:1:3) System suitability solution: Aprotinin solution that contains 5 USP Aprotinin Units/mL with 2% aprotinin oligomers [NOTEThis solution can be obtained by heating lyophilized aprotinin at 112 for 2 h and dissolving the solid at the specified concentration in water.] Sample solution: 5 USP Aprotinin Units/mL of Aprotinin Chromatographic system (see Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: Series of three 7.8-mm 30-cm columns; packing L33 Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: System suitability solution [NOTEThe relative retention times of dimer and aprotinin are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.3 between the dimer peak and the aprotinin peak D
Dry 100 mg in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 6.0% of its weight. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.14 USP Endotoxin Unit/USP Aprotinin Unit. Use a solution that contains 6 USP Aprotinin Units/mL. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: For lyophilized powder, preserve in tight containers, and store in a cold place. Protect from light. For bulk solution, preserve in tight containers at a temperature not exceeding 25. Avoid freezing. LABELING: The labeling states the source of material and the number of Kallikrein Inhibition Units/mg or the number of Kallikrein Inhibition Units/mL. USP REFERENCE STANDARDS 11 USP Aprotinin RS USP Aprotinin System Suitability RS USP Endotoxin RS USP Trypsin Crystallized RS
Aprotinin Injection
(Comment on this Monograph)id=m5745=Aprotinin Injection=A-Monos.pdf) DEFINITION Aprotinin Injection is a sterile solution of Aprotinin in Water for Injection that also contains sodium chloride. One USP Aprotinin Unit is equivalent to 1800 Kallikrein Inhibition Units (K.I.U.). It contains NLT 90.0% and NMT 110.0% of the potency stated on the label, expressed in Kallikrein Inhibition Units per mL. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the test for Limit of N-pyroglutamyl-aprotinin and related compounds. B. The determination of activity by the Assay is based on the specific inhibition of trypsin. ASSAY PROCEDURE 0.0015 M Borate buffer: Transfer 0.93 g of boric acid into a 1000-mL volumetric flask, dissolve in 900 mL of water, adjust with 5 N sodium hydroxide to a pH of 8.0, and dilute with water to volume. Transfer 100 mL of this solution into a 1000-mL volumetric flask, and dilute with water to volume. Sample solution: 1.67 USP Aprotinin Units/mL (about 0.6 mg/mL) of Aprotinin in 0.0015 M Borate buffer Trypsin solution: 4300 USP Trypsin Units/mL of USP Trypsin Crystallized RS in 0.001 N hydrochloric acid [NOTE Use a freshly prepared solution, and keep in ice water.] Trypsin and aprotinin solution: To 4.0 mL of the Trypsin solution, add 1.0 mL of the Sample solution. Dilute immediately with 0.0015 M Borate buffer to 40.0 mL. Allow to stand at room temperature for 10 min, and then keep in ice water. [NOTEUse within 6 h of preparation.] Dilute trypsin solution: Trypsin solution and 0.0015 M Borate buffer (0.5:9.5). Allow to stand at room temperature for 10 min, and then keep in ice water. Substrate solution: 6.9 mg/mL of N-benzoyl-L-arginine ethyl ester hydrochloride in water [NOTEUse within 2 h.] Analysis: Mix 9.0 mL of 0.0015 M Borate buffer and 1.0 mL of Substrate solution in a jacketed-glass vessel with a capacity of 30 mL and that contains a stirring device. The lid of the
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Tailing factor: NMT 2.5 for the aprotinin peak Retention time: 24.525.5 min for aprotinin Analysis Sample: Sample solution Calculate the percentage of each oligomer peak in the chromatogram: Result = (ri/rT) 100 = response of each peak having a retention time less than that of aprotinin monomer = sum of the responses of all peaks rT Acceptance criteria: Sum of all oligomers is NMT 1.5% ri SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements PH 791: 4.56.5 INJECTIONS 1: Meets the requirements LIMIT OF N-PYROGLUTAMYL-APROTININ AND RELATED COMPOUNDS Solution A: 3.52 mg/mL of monobasic potassium phosphate and 7.26 mg/mL of dibasic sodium phosphate in water Solution B: 3.52 mg/mL of monobasic potassium phosphate, 7.26 mg/mL of dibasic sodium phosphate, and 66.07 mg/mL of ammonium sulfate dissolved in water System suitability solution: 5 USP Aprotinin Units/mL of USP Aprotinin System Suitability RS in Solution A Sample solution: 5 USP Aprotinin Units/mL of Aprotinin in Solution A Chromatographic system (see Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 7.5-mm 7.5-cm; packing L52 Temperature: 40 (constant temperature) Flow rate: 1 mL/min
Official Monographs / Arginine 271

Acceptance criteria N-pyroglutamyl-aprotinin: NMT 1.0% Any other impurity: NMT 0.5% Sum of all unknown impurities: NMT 1.0% BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.14 USP Endotoxin Units/USP Aprotinin Unit. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, store at up to 25, and avoid freezing. USP REFERENCE STANDARDS 11 USP Aprotinin RS USP Aprotinin System Suitability RS USP Endotoxin RS USP Trypsin Crystallized RS
Arginine
(Comment on this Monograph)id=m5840=Arginine=AMonos.pdf)
L-Arginine
C6H14N4O2
[74-79-3].
174.20
DEFINITION Arginine contains NLT 98.5% and NMT 101.5% of (C6H14N4O2), as L-arginine, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 80 mg of Arginine Analysis: Dissolve the Sample in a mixture of 3 mL of formic acid and 50 mL of glacial acetic acid in a 125-mL flask. Titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 8.710 mg of C6H14N4O2. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.3% CHLORIDE AND SULFATE, Chloride 221: NMT 500 ppm. A 1.0-g portion shows no more chloride than corresponds to 0.70 mL of 0.020 N hydrochloric acid. CHLORIDE AND SULFATE, Sulfate 221: NMT 300 ppm. A 1.0g portion shows no more sulfate than corresponds to 0.30 mL of 0.020 N sulfuric acid. IRON 241: NMT 30 ppm HEAVY METALS 231, Method I: NMT 15 ppm Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 0.05 mg/mL of USP L-Arginine RS in 0.1 N hydrochloric acid [NOTEThis solution has a concentration equivalent to 0.5% of that of the Sample solution.] Sample solution: 10 mg/mL of Arginine in 2 N hydrochloric acid
Injection size: 40 L System suitability Sample: System suitability solution [NOTEThe relative retention times for N-pyroglutamylaprotinin and aprotinin are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.0 between N-pyroglutamyl-aprotinin and aprotinin Tailing factor: NMT 2.0 for the aprotinin peak Retention time: 1720 min for aprotinin Analysis Sample: Sample solution Calculate the percentage of each impurity peak in the chromatogram: Result = (ri/rT) 100 ri rT = response of each impurity peak = sum of the responses of all peaks from the Sample solution
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until the silver chloride flocculates and the mixture acquires a faint pink color, using 1 mL of dichlorofluorescein TS as indicator. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride. Acceptance criteria: 16.5%17.1% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Sulfate 221: NMT 300 ppm. A 1.6g portion shows no more sulfate than corresponds to 0.50 mL of 0.020 N sulfuric acid. HEAVY METALS, Method I 231: NMT 20 ppm. Proceed as directed except to dissolve 1.0 g in 20 mL of water, add 2 mL of 1 N acetic acid, and dilute with water to 25 mL. Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 0.05 mg/mL of USP Arginine Hydrochloride RS in water. [NOTEThis solution has a concentration equivalent to 0.5% of that of the Sample solution.] Sample solution: 10 mg/mL of Arginine Hydrochloride in water System suitability solution: 0.4 mg/mL each of USP Arginine Hydrochloride RS and USP L-Lysine Hydrochloride RS in water Spray reagent: 2 mg/mL of ninhydrin in butyl alcohol and 2 N acetic acid (19:1) Application volume: 5 L Developing solvent system: Isopropyl alcohol and ammonium hydroxide (7:3) Analysis Samples: Standard solution, Sample solution, and System suitability solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Dry the plate between 100 and105 until the ammonia disappears completely. Spray with Spray reagent, and heat between 100 and 105 for 15 min. Examine the plate under white light. The chromatogram from the System suitability solution exhibits two clearly separated spots. Any secondary spot from the Sample solution is not larger or more intense than the principal spot from the Standard solution. Acceptance criteria Individual impurities: NMT 0.5% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +21.4 to +23.6 (t = 20) Sample solution: 80 mg/mL in 6 N hydrochloric acid LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.2% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Arginine Hydrochloride RS USP L-Lysine Hydrochloride RS
System suitability solution: 0.4 mg/mL each of USP LArginine RS and USP L-Lysine Hydrochloride RS in 0.1 N hydrochloric acid Application volume: 5 L Spray reagent: 2 mg/mL of ninhydrin in a mixture of butyl alcohol and 2 N acetic acid (19:1) Application volume: 5 L Developing solvent system: Isopropyl alcohol and ammonium hydroxide (7:3) Analysis Samples: Standard solution, Sample solution, and System suitability solution Proceed as directed under Chromatography 621, ThinLayer Chromatography. Dry the plate between 100 and 105 until the ammonia disappears completely. Spray with Spray reagent, and heat between 100 and 105 for about 15 min. Examine the plate under white light. The chromatogram obtained from the System suitability solution exhibits two clearly separated spots. Any secondary spot from the Sample solution is not larger or more intense than the principal spot from the Standard solution. Acceptance criteria Individual impurities: NMT 0.5% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +26.3 to +27.7 Sample solution: 80 mg/mL in 6 N hydrochloric acid LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP L-Arginine RS USP L-Lysine Hydrochloride RS
Arginine Hydrochloride
(Comment on this Monograph)id=m5850=Arginine Hydrochloride=A-Monos.pdf)
L-Arginine monohydrochloride; L-(+)-Arginine monohydrochloride
C6H14N4O2 HCl
210.66 [1119-34-2].
DEFINITION Arginine Hydrochloride contains NLT 98.5% and NMT 101.5% of C6H14N4O2 HCl, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 100 mg of Arginine Hydrochloride Analysis: Dissolve Sample in 3 mL of 98% formic acid and 50 mL of glacial acetic acid. Add 6 mL of mercuric acetate TS and titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 10.53 mg of C6H14N4O2 HCl. Acceptance criteria: 98.5%101.5% OTHER COMPONENTS CHLORIDE CONTENT Sample: 350 mg [NOTEUse a porcelain casserole.] Analysis: Add 140 mL of water and 1 mL of dichlorofluorescein TS. Titrate with 0.1 N silver nitrate VS
Arginine Hydrochloride Injection

(Comment on this Monograph)id=m5880=Arginine Hydrochloride Injection=A-Monos.pdf) DEFINITION Arginine Hydrochloride Injection is a sterile solution of Arginine Hydrochloride in Water for Injection. It contains NLT 9.5% and
USP 32
NMT 10.5% of C6H14N4O2 HCl. It contains no antimicrobial agents. [NOTEThe chloride ion content of Arginine Hydrochloride Injection is approximately 475 mEq/L.] IDENTIFICATION A. Transfer 1 mL of the Injection to a 200-mL volumetric flask, and dilute with water to volume. To 1 mL of this dilution add 2 mL of a solution of 0.02% 8-hydroxyquinoline in 3 N sodium hydroxide, and add 1 mL of 0.1% Nbromosuccinimide solution. Acceptance criteria: An orange color is produced. B. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Color reagent: Dissolve 28.0 g of potassium hydroxide and 2.0 g of potassium sodium tartrate in 100 mL of water. Cool, and add, in the order named, 100 mg of 2,4dichloro-1-naphthol, 180 mL of alcohol, and 20.0 mL of 0.475% sodium hypochlorite solution. Mix by swirling, and allow to stand at room temperature for 1 h before using. This Color reagent may be stored in a glass-stoppered bottle, in a refrigerator, for 2 months. Standard solution: 40 g/mL of USP Arginine Hydrochloride RS in water Sample solution: Equivalent to 40 g/mL of arginine hydrochloride, from a volume of Injection in water Analysis: Transfer 2.0-mL portions of the Sample solution and the Standard solution, respectively, to separate flasks, and treat each as follows. Add 2.0 mL of potassium iodide solution (3 in 1000), and allow to stand for 15 min. Add 6.0 mL of Color reagent, and allow to stand for 15 min. Add 2.0 mL of sodium hypochlorite solution (19 in 10,000), and allow to stand for 15 min. Spectrometric conditions Cell: 1 cm Analytical wavelength: 520 nm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H14N4O2 HCl in each mL of the Injection taken: Result = (AU/AS) (CS/CU) 100 absorbances of the Sample solution absorbances of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 9.5%10.5% AU AS CS CU SPECIFIC TESTS PH 791: 5.06.5 OTHER REQUIREMENTS: Meets the requirements under Injections 1 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.01 USP Endotoxin Unit/mg of arginine hydrochloride. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type II glass. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, or where the label states that the Injection is not for direct injection but is to be diluted before use, the label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Arginine Hydrochloride RS = = = =
Official Monographs / Arsanilic 273

USP Endotoxin RS
Arsanilic Acid
(Comment on this Monograph)id=m5940=Arsanilic Acid=AMonos.pdf)
C6H8AsNO3 p-Aminobenzenearsonic acid [98-50-0]. DEFINITION Arsanilic Acid contains NLT 98.0% and NMT 102.0% of C6H8AsNO3, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K
217.05
ASSAY PROCEDURE Sample solution: 125 mg of Arsanilic Acid transferred to a 50-mL conical flask Analysis: Add 10.0 mL of a mixture of sulfuric acid, nitric acid, and perchloric acid (1000:50:50) and several glass beads. Digest on a hot plate for 1 h, increasing the temperature of the hot plate in steps until a ring of sulfuric acid rises into the neck of the flask. Allow to cool, to the colorless solution add 400 mg of hydrazine sulfate, and heat the flask vigorously on a hot plate until a ring of sulfuric acid rises into the neck of the flask. Allow to cool, and wash down the rim, neck, and insides of the flask with 1 mL of water. Heat the flask again until a ring of sulfuric acid rises into the neck of the flask. Allow to cool, and transfer the colorless solution, with the aid of 80 mL of water, to a 125mL conical flask. Add 10 mL of hydrochloric acid and several drops of 0.002 M potassium iodide, cool to 05 and titrate with 0.1 N potassium permanganate VS to a pale pink endpoint, maintaining a temperature of 05 during the titration. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N potassium permanganate is equivalent to 10.852 mg of C6H8AsNO3. Acceptance criteria: 98.0%102.0% IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF O-ARSANILIC ACID Mobile phase: 4.04 g of monobasic potassium phosphate in 985 mL of water. Add 2 mL of phosphoric acid. Add 10 mL of methanol. Standard solution: Transfer 67 mg of o-arsanilic acid to a 100-mL volumetric flask, add 65 mg of warm (7080) water, and shake or sonicate to dissolve. Allow to cool, and dilute with water to volume. Dilute a portion of this solution with water to obtain a solution having a known concentration of 0.0012 mg/mL of o-arsanilic acid. Sample solution: Transfer 50 mg of Arsanilic Acid to a 50mL volumetric flask, add 30 mL of warm water, and shake or sonicate to dissolve. Allow to cool, and dilute with water to volume. Chromatographic system (See Chromatography, 621 System Suitability.)
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W
USP 32
= weight of Arsanilic Acid in the Sample solution (mg) Acceptance criteria: NMT 0.045% SPECIFIC TESTS LOSS ON DRYING 731: Dry it in a vacuum at 80 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Arsanilic Acid RS
Mode: LC Detector: UV 242nm Column: 4.6-mm 15-cm; 5-m base-deactivated packing L1 Temperature: 30 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.5% Capacity factor: 2.83.8 for the o-arsanilic acid peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of o-arsanilic acid in the portion of C6H8AsNO3 taken: Result = 5000 (CS/W) (rU/rS) = concentration of o-arsanilic acid in the Standard solution (mg/mL) W = weight of Arsanilic Acid in the Sample solution (mg) = o-arsanilic acid peak response from the Sample rU solution rS = o-arsanilic acid peak response from the Standard solution Acceptance criteria: NMT 0.12% PROCEDURE 2: LIMIT OF ANILINE Mobile phase: 7.76 g of monobasic potassium phosphate in 950 mL of water. Add 50 mL of methanol. Standard solution: Transfer 176 mg of aniline to a 25-mL volumetric flask, add 1 mL of methanol, swirl, then add 15 mL of water, and shake to dissolve. Dilute with water to volume. Dilute a portion of this solution to obtain a solution having a known concentration of 0.00045 mg/mL of aniline. Sample solution: Transfer 50 mg of Arsanilic Acid to a 50mL volumetric flask, add 30 mL of warm (7080) water, and shake or sonicate to dissolve. Allow to cool, and dilute with water to volume. Chromatographic system (See Chromatography, 621 System Suitability.) Mode: LC Detector: UV 235nm Column: 4.6-mm 15-cm; 5-m base-deactivated packing L1 Temperature: 30 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Capacity factor: 2.33.3 for the aniline peak Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of aniline in the portion of Arsanilic Acid taken: CS Result = (rU/rS) (CS/W) 5000 rU rS CS = aniline peak response from the Sample solution = aniline peak response from the Standard solution = concentration of aniline in the Standard solution (mg/mL)
Ascorbic Acid
(Comment on this Monograph)id=m6030=Ascorbic Acid=AMonos.pdf)
C6H8O6 L-Ascorbic acid; 1-Deoxy-L-gulofuranose-2-ene-1-one [50-81-7].
176.12
DEFINITION Ascorbic Acid contains NLT 99.0% and NMT 100.5% of C6H8O6 IDENTIFICATION A. INFRARED ABSORPTION 197K B. A 20 mg/mL solution reduces alkaline cupric tartrate TS slowly at room temperature but more readily upon heating. ASSAY PROCEDURE Sample solution: 3.2 mg/mL of Ascorbic Acid, in a mixture of 2 N sulfuric acid and water (1:4) Analysis: Titrate 125 mL of the Sample solution at once with 0.1 N iodine VS, using 3 mL of starch TS as an indicator. Each mL of 0.1 N iodine is equivalent to 8.806 mg of C 6 H8 O6 . Acceptance criteria: 99.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS 231: NMT 20 ppm Sample solution: 40 mg/mL in water SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +20.5 to +21.5, the optical rotation being measured immediately, following the preparation of the solution Sample solution: 100 mg/mL, in carbon dioxide-free water ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Ascorbic Acid RS
USP 32
Official Monographs / Ascorbic 275

LIMIT OF OXALATE: Dilute a volume of Injection, equivalent to 50 mg of ascorbic acid, with water to 5 mL. Add 0.2 mL of acetic acid and 0.5 mL of calcium chloride TS: no turbidity is produced in 1 min. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in light-resistant, singledose containers, preferably of Type I or Type II glass. LABELING: In addition to meeting the requirements under Injections 1, Labeling, fused-seal containers of the Injection in concentrations of 250 mg/mL and greater are labeled to indicate that since pressure may develop on long storage, precautions should be taken to wrap the container in a protective covering while it is being opened. USP REFERENCE STANDARDS 11 USP Ascorbic Acid RS USP Endotoxin RS
Ascorbic Acid Injection

(Comment on this Monograph)id=m6050=Ascorbic Acid Injection=A-Monos.pdf) DEFINITION Ascorbic Acid Injection is a sterile solution, in Water for Injection, of Ascorbic Acid prepared with the aid of Sodium Hydroxide, Sodium Carbonate, or Sodium Bicarbonate. It contains NLT 90.0% and NMT 110.0% of the labeled amount of ascorbic acid (C6H8O6). IDENTIFICATION A. To a volume of Injection, equivalent to 40 mg of ascorbic acid, add 4 mL of 0.1 N hydrochloric acid, then add 4 drops of methylene blue TS, and warm to 40: the deep blue color becomes appreciably lighter or is completely discharged within 3 min. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, obtained as directed in the Assay. C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements ASSAY PROCEDURE Mobile phase: Dissolve 15.6 g of dibasic sodium phosphate and 12.2 g of monobasic potassium phosphate in 2000 mL of water, and adjust with phosphoric acid to a pH of 2.5 0.05. Standard solution: 0.5 mg/mL of USP Ascorbic Acid RS in Mobile phase [NOTERefrigerate and store protected from light until use. The solution is stable for at least 24 h. Inject within 3 h after removal from the refrigerator.] Sample solution: 0.5 mg/mL from Injection in Mobile phase [NOTERefrigerate and store protected from light until use. The solution is stable for at least 24 h. Inject within 3 h after removal from the refrigerator.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 245 nm Column: 6-mm 150-cm; packing L39 Flow rate: 0.6 mL/min Injection size: 4 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3500 theoretical plates Tailing factor: NMT 1.6 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H8O6 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ascorbic Acid RS in the Standard solution (mg/mL) = nominal concentration of ascorbic acid in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 5.57.0 OTHER REQUIREMENTS: It meets the requirements under Injections 1. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.2 USP Endotoxin Units/mg of ascorbic acid. rU rS CS
Ascorbic Acid Oral Solution

(Comment on this Monograph)id=m6070=Ascorbic Acid Oral Solution=A-Monos.pdf) DEFINITION Ascorbic Acid Oral Solution is a solution of Ascorbic Acid in a hydroxylic organic solvent or an aqueous mixture thereof. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C6H8O6. IDENTIFICATION A. To a volume of Oral Solution equivalent to 40 mg of ascorbic acid add 4 mL of 0.1 N hydrochloric acid, then add 4 drops of methylene blue TS, and warm to 40: the deep blue color becomes appreciably lighter or is completely discharged within 3 min. B. To a volume of Oral Solution equivalent to 20 mg of ascorbic acid add 15 mL of trichloroacetic acid solution (1 in 20), add 200 mg of activated charcoal, shake the mixture vigorously for 1 min, and pass through a small fluted filter, returning the filtrate, if necessary, until clear. To 5 mL of the filtrate add 1 drop of pyrrole, agitate gently until dissolved, then heat in a bath at 50: a blue color develops. ASSAY PROCEDURE Sample solution: Transfer an equivalent to 50 mg of ascorbic acid from a volume of the Oral Solution to a 100mL volumetric flask, previously diluted with water if necessary. Add 20 mL of metaphosphoric-acetic acids TS, and dilute with water to volume. Measure a volume of the dilution, equivalent to 2 mg of ascorbic acid, into a 50 mL conical flask, and add 5 mL of metaphosphoric-acetic acids TS. Analysis: Titrate with standard dichlorophenolindophenol solution until a rose-pink color persists for at least 5 s. Correct for the volume of the dichlorophenolindophenol solution consumed by a mixture of 5.5 mL of metaphosphoric-acetic acids TS and 15 mL of water. From the ascorbic acid equivalent of the standard dichlorophenolindophenol solution, calculate the ascorbic acid content in each mL of the Oral Solution. Acceptance criteria: If present, 90.0%110.0% OTHER COMPONENTS ALCOHOL DETERMINATION, Method I 611: 90.0%110.0%
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Ascorbic / Official Monographs
USP 32
LABELING: Label Oral Solution that contains alcohol to state the alcohol content.
Aspartic Acid
(Comment on this Monograph)id=m6198=Aspartic Acid=AMonos.pdf)
Ascorbic Acid Tablets

(Comment on this Monograph)id=m6090=Ascorbic Acid Tablets=A-Monos.pdf) DEFINITION Ascorbic Acid Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C6H8O6. IDENTIFICATION [NOTEThe Sample solution is prepared as follows.] Sample solution: Triturate a quantity of finely powdered Tablets with sufficient diluted alcohol to make approximately the equivalent of a 1-in-50 solution of ascorbic acid, filter, and proceed with the following tests. A. A portion of the filtrate reduces alkaline cupric tartrate TS slowly at room temperature but more readily upon heating. B. To 2 mL of the filtrate add 4 drops of methylene blue TS, and warm to 40: the deep blue color becomes appreciably lighter or is completely discharged within 3 min. C. To 1 mL of the filtrate add 15 mL of a solution of trichloroacetic acid (1 in 20), add 200 mg of activated charcoal, shake the mixture vigorously for 1 min, and pass through a small fluted filter, returning the filtrate, if necessary, until clear. To 5 mL of the filtrate add 1 drop of pyrrole, and agitate gently until dissolved, then heat in a bath at 50: a blue color develops. ASSAY PROCEDURE Sample solution: Transfer NLT 20 Tablets to a 1000-mL volumetric flask containing 250 mL of metaphosphoricacetic acids TS. Analysis: Insert the stopper in the flask, and shake by mechanical means for 30 min or until the tablets have disintegrated completely. Dilute with water to volume. Transfer a portion of the solution to a centrifuge tube, and centrifuge until a clear supernatant is obtained. Quantitatively dilute the clear supernatant with water, if necessary, to obtain a solution containing 500 g/mL of ascorbic acid. Pipet 4 mL of the solution, equivalent to 2 mg of ascorbic acid, into a 50-mL conical flask. Add 5 mL of metaphosphoric-acetic acids TS and titrate with standard dichlorophenolindophenol solution until a rose-pink color persists for at least 5 s. Correct for the volume of the dichlorophenolindophenol solution consumed by a mixture of 5.5 mL of metaphosphoric-acetic acids TS and 15 mL of water. From the ascorbic acid equivalent of the standard dichlorophenolindophenol solution, calculate the content of ascorbic acid in each Tablet. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Analysis: Determine the amount of C6H8O6 dissolved, using the Analysis set forth in the Assay and conducting the Analysis without delay, making any necessary modifications. Tolerances: NLT 75% (Q) of the labeled amount of C6H8O6 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.
C4H7NO4 L-Aspartic acid [56-84-8]. DEFINITION Aspartic Acid contains NLT 98.5% and NMT 101.5% of C4H7NO4, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K
133.10
ASSAY PROCEDURE Sample: 0.1 g of Aspartic Acid Analysis: In a 125-mL flask, dissolve in 50 mL of carbon dioxide-free water, heating slightly if necessary. Cool, add 0.1 mL of bromothymol blue TS, and titrate with 0.1 N sodium hydroxide VS until the color changes from yellow to blue. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N sodium hydroxide is equivalent to 13.31 mg of C4H7NO4. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221: Dissolve 0.7 g in 10 mL of diluted nitric acid, and dilute with water to make 15 mL. Acceptance criteria: The solution shows no more chloride than corresponds to 0.20 mL of 0.020 N hydrochloric acid (0.02%). CHLORIDE AND SULFATE, Sulfate 221: Dissolve 0.8 g in 4 mL of hydrochloric acid, and dilute with water to make 15 mL. Acceptance criteria: The solution shows no more sulfate than corresponds to 0.25 mL of 0.020 N sulfuric acid (0.03%). IRON 241: NMT 10 ppm HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel System suitability solution: 10 mg each of USP Aspartic Acid RS and glutamic acid in 2 mL of ammonia TS, dilute with water to 25.0 mL Sample solution: Transfer 0.1 g of Aspartic Acid to a 10mL volumetric flask, dissolve in 2 mL of 17% ammonia solution (prepared by diluting ammonium hydroxide, 6 in 10), and dilute with water to volume. Standard solution: Transfer 5 mg of USP Aspartic Acid RS to a 100-mL volumetric flask, dissolve in 2 mL of 17% ammonia solution (prepared by diluting ammonium hydroxide, 6 in 10), and dilute with water to volume. Application volume: 5 L Developing solvent system: Butyl alcohol, glacial acetic acid, and water (3:1:1) Spray reagent: 2 mg/mL of ninhydrin in a mixture of butyl alcohol and 2 N acetic acid (19:1)
USP 32
Analysis Samples: System suitability solution, Sample solution, and Standard solution Analysis: Proceed as directed for Chromatography 621, Thin-Layer Chromatography, except to dry the plate at 80 for 30 min, spray with Spray reagent, and heat at 80 for 30 min. Examine the plate under white light. The chromatogram obtained from the System suitability solution exhibits two clearly separated spots, and no secondary spot in the chromatogram of the Sample solution is larger or more intense than the principal spot in the chromatogram of the Standard solution. Acceptance criteria Individual impurities: NMT 0.5% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +24.0 to +26.0, at 20 Sample solution: 80 mg/mL, in 6 N hydrochloric acid LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, protected from light. USP REFERENCE STANDARDS 11 USP Aspartic Acid RS
Official Monographs / Aspirin 277

Aspirin
(Comment on this Monograph)id=m6240=Aspirin=AMonos.pdf)
C9H8O4 Benzoic acid, 2-(acetyloxy)-; Salicylic acid acetate [50-78-2].
180.16
DEFINITION Aspirin contains NLT 99.5% and NMT 100.5% of C9H8O4, calculated on the dried basis. IDENTIFICATION A. Heat it with water for several min, cool, and add 1 or 2 drops of ferric chloride TS: a violet-red color is produced. B. INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 1.5 g of Aspirin in a flask Analysis: Add 50.0 mL of 0.5 N sodium hydroxide VS, and boil the mixture gently for 10 min. Add phenolphthalein TS, and titrate the excess sodium hydroxide with 0.5 N sulfuric acid VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.5 N sodium hydroxide is equivalent to 45.04 mg of C9H8O4.
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.05% CHLORIDE AND SULFATE 221 Sample: 1.5 g Analysis: Boil Sample with 75 mL of water for 5 min, cool, add sufficient water to restore the original volume, and filter. Acceptance criteria: A 25-mL portion of the filtrate shows no more chloride than corresponds to 0.10 mL of 0.020 N hydrochloric acid (140 ppm). SULFATE Sample: 6.0 g Analysis: Dissolve Sample in 37 mL of acetone, and add 3 mL of water. Titrate potentiometrically with 0.02 M lead perchlorate, prepared by dissolving 9.20 g of lead perchlorate in water to make 1000 mL of solution, using a pH meter capable of a minimum reproducibility of 0.1 mV (see pH 791), and equipped with an electrode system consisting of a lead-specific electrode and a silversilver chloride reference glass-sleeved electrode containing a solution of tetraethylammonium perchlorate in glacial acetic acid (1 in 44) (see Titrimetry 541). [NOTEAfter use, rinse the lead-specific electrode with water, drain the reference electrode, flush with water, rinse with methanol, and allow to dry.] Acceptance criteria: NMT 1.25 mL of 0.02 M lead perchlorate is consumed (NMT 400 ppm). HEAVY METALS Sample: 2 g Analysis: Dissolve in 25 mL of acetone, and add 1 mL of water. Add 1.2 mL of thioacetamideglycerin base TS and 2 mL of pH 3.5 Acetate Buffer (see Heavy Metals 231), and allow to stand for 5 min. Acceptance criteria: Any color produced is not darker than that of a control made with 25 mL of acetone and 2 mL of Standard Lead Solution (see Heavy Metals 231), treated in the same manner (NMT 1 ppm). Organic Impurities PROCEDURE 1: LIMIT OF FREE SALICYLIC ACID Sample: 2.5 g Analysis: Dissolve in sufficient alcohol to make 25.0 mL. To each of two matched color-comparison tubes add 48 mL of water and 1 mL of a freshly prepared, diluted ferric ammonium sulfate solution (prepared by adding 1 mL of 1 N hydrochloric acid to 2 mL of ferric ammonium sulfate TS, and diluting with water to 100 mL). Into one tube pipet 1 mL of a standard solution of salicylic acid in water, containing 0.10 mg/mL of salicylic acid. Into the second tube pipet 1 mL of the solution (1 in 10) of Aspirin. Mix the contents of each tube. Acceptance criteria: After 30 s, the color in the second tube is not more intense than that in the tube containing the salicylic acid (NMT 0.1%). PROCEDURE 2: READILY CARBONIZABLE SUBSTANCES TEST 271 Sample solution: 100 mg/mL in sulfuric acid TS Acceptance criteria: The solution has no more color than Matching Fluid Q. PROCEDURE 3: SUBSTANCES INSOLUBLE IN SODIUM CARBONATE TS: A solution of 500 mg in 10 mL of warm sodium carbonate TS is clear.
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USP 32
SPECIFIC TESTS LOSS ON DRYING 731: Dry it over silica gel for 5 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS
Aspirin Boluses
(Comment on this Monograph)id=m6245=Aspirin Boluses=AMonos.pdf) DEFINITION Aspirin Boluses contain NLT 90.0% and NMT 110.0% of the labeled amount of aspirin (C9H8O4). IDENTIFICATION A. PROCEDURE Analysis: Crush 1 Bolus. Boil a portion of the powder, equivalent to 300 mg of aspirin, with 50 mL of water. Cool, and add a drop of ferric chloride TS. Acceptance criteria: A violet-red color is produced. B. The retention time of the aspirin peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 2 mg/mL of sodium 1-heptanesulfonate in acetonitrile and water (3:17). Adjust with glacial acetic acid to a pH of 3.4. Diluent: Acetonitrile and formic acid (99:1) Standard solution: 0.4 mg/mL of USP Aspirin RS and 0.01 mg/mL of USP Salicylic Acid RS in Diluent Sample solution: Finely powder NLT 10 Boluses. Transfer a portion of the powder to an appropriate volumetric flask and dilute with Diluent to prepare a solution nominally equivalent to 4 mg/mL. Stir the solution by mechanical means for 15 min. Pass a portion of this solution through a filter having a 0.5-m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and aspirin are 0.6 and 1.0, respectively.] Suitability requirements Relative standard deviation: NMT 2.0% for the aspirin peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Boluses taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Aspirin RS in the Standard solution (mg/mL) = nominal concentration of aspirin in the Sample solution (mg/mL)
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.5 M phosphate buffer, pH 7.4; 900 mL Apparatus 2: 75 rpm Time: 45 min Detector: UV absorption at the wavelength of the isosbestic point of aspirin and salicylic acid at 265 2 nm Diluent: Acetonitrile and formic acid (99:1) Sample solutions: Filtered portions of the solution under test, suitably diluted with Diluent Standard solution: USP Aspirin RS in Medium at a known concentration [NOTEPrepare the solution at the time of use.] Analysis: Determine the amount of C9H8O4 dissolved by UV absorption performed on the Sample solution, in comparison to a Standard solution having a known concentration of USP Aspirin RS. Tolerances: NLT 80% (Q) of the labeled amount of C9H8O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF SALICYLIC ACID Analysis: Using the chromatograms of the Standard solution and the Sample solution, obtained as directed in the Assay, calculate the percentage of C7H6O3 in the portion of Boluses taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) = concentration of aspirin in the portion of Boluses CU taken for the Sample solution, as determined in the Assay (mg/mL) Acceptance criteria: NMT 0.3% rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label Boluses to indicate that they are for veterinary use only. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
Aspirin Capsules
(Comment on this Monograph)id=m6250=Aspirin Capsules=AMonos.pdf) DEFINITION Aspirin Capsules contain NLT 93.0% and NMT 107.0% of the labeled amount of aspirin (C9H8O4). [NOTECapsules that are enteric-coated or the contents of which are enteric-coated meet the requirements for Aspirin Delayed-Release Capsules.] IDENTIFICATION A. PROCEDURE Sample: 1 Tablet Analysis: Crush and boil it with 50 mL of water for 5 min, cool, and add 1 or 2 drops of ferric chloride TS.
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Acceptance criteria: A violet-red color is produced. B. INFRARED ABSORPTION 197K Sample: A quantity of the contents of Capsules, equivalent to 500 mg of aspirin Analysis: Shake Sample with 10 mL of alcohol for several min. Centrifuge the mixture. Pour off the clear supernatant and evaporate it to dryness. Dry the residue in a vacuum at 60 for 1 h. Acceptance criteria: Meet the requirements ASSAY PROCEDURE [NOTEUse chloroform recently saturated with water.] Diluent: A solution (1 in 100) of glacial acetic acid in chloroform Standard stock solution: Transfer 50 mg of USP Aspirin RS to a 50-mL volumetric flask, add 0.5 mL of glacial acetic acid, and add chloroform to volume. Standard solution: 50 g/mL of USP Aspirin RS from Standard stock solution diluted with Diluent Chromatographic column: Proceed as directed under Chromatography 621, Column Partition Chromatography, packing a chromatographic tube with a mixture of 3 g of Solid Support and 2 mL of freshly prepared sodium bicarbonate solution (1 in 12). Sample solution: Remove, as completely as possible, the contents of NLT 20 Capsules. Mix the combined contents, and transfer a quantity of the powder, equivalent to 50 mg of aspirin, to a 50-mL volumetric flask containing 1 mL of a solution (1 in 50) of hydrochloric acid in methanol, and add chloroform to volume. Transfer 5.0 mL of this solution to the column, wash with 5 mL and then with 25 mL of chloroform, and discard the washings. Elute into a 100-mL volumetric flask with 10 mL of a solution (1 in 10) of glacial acetic acid in chloroform and then with 85 mL of a solution (1 in 100) of glacial acetic acid in chloroform, and dilute with the latter solvent to volume. Spectrometric conditions Mode: UV Analytical wavelength: 280 nm Cell: 1 cm Blank: Chloroform Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Capsules taken: Result = (AU/AS) (CS/CU) 100 = absorbances of the Sample solution = absorbances of the Standard solution = concentration of USP Aspirin RS in the Standard solution (g/mL) = nominal concentration of aspirin in the Sample CU solution (mg/mL) Acceptance criteria: 93.0%107.0% AU AS CS PERFORMANCE TESTS DISSOLUTION 711 0.05 M acetate buffer: Mix 2.99 g of sodium acetate trihydrate and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05. Medium: 0.05 M acetate buffer; 500 mL Apparatus 1: 100 rpm Time: 30 min Standard solution: USP Aspirin RS in Medium [NOTEPrepare the Standard solution at the time of use. An amount of alcohol not to exceed 1% of the total volume of the Standard solution may be used to bring the Reference Standard into solution prior to dilution with Medium.]

Sample solution: Sample per Dissolution 711. Dilute with Medium, if necessary, and filter. Analysis Samples: Standard solution and Sample solution Determine the amount of C9H8O4 dissolved from UV absorbances at the wavelength of the isosbestic point of aspirin and salicylic acid at 265 2 nm of the Sample solution in comparison with a Standard solution having a known concentration. Tolerances: NLT 80% (Q) of the labeled amount of C9H8O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Ferric chloride-urea reagent: Dissolve by swirling, without the aid of heat, 60 g of urea in a mixture of 8 mL of ferric chloride solution (6 in 10) and 42 mL of 0.05 N hydrochloric acid. Adjust the resulting solution, if necessary, with 6 N hydrochloric acid to a pH of 3.2. Standard solution: Transfer 75.0 mg of salicylic acid, previously dried over silica gel for 3 h, to a 100-mL volumetric flask, and add chloroform to volume. Transfer 10.0 mL of this solution to a second 100-mL volumetric flask, and dilute with chloroform to volume. Transfer 10.0 mL of this last solution to a 50-mL volumetric flask containing 10 mL of methanol, 2 drops of hydrochloric acid, and 10 mL of a solution (1 in 10) of glacial acetic acid in ether, and dilute with chloroform to volume. Chromatographic column: Proceed as directed under Chromatography 621, Column Partition Chromatography, packing a chromatographic tube with two segments of packing material. The lower segment is a mixture of 1 g of Solid Support and 0.5 mL of 5 M phosphoric acid, and the upper segment is a mixture of 3 g of Solid Support and 2 mL of freshly prepared Ferric chloride-urea reagent. Sample solution: Weigh a portion of the contents of the Capsules, as determined by the Assay, equivalent to 100 mg of aspirin, mix with 10 mL of chloroform by stirring for 3 min, and then transfer to the chromatographic column with the aid of a few mL of chloroform. Pass 50 mL of chloroform through the column, rinse the tip of the chromatographic tube with chloroform, and discard the eluate. Prepare as a receiver a 50-mL volumetric flask containing 10 mL of methanol and 2 drops of hydrochloric acid, and elute any salicylic acid from the column by passing 10 mL of a solution (1 in 10) of glacial acetic acid in ether that has been recently saturated with water, followed by 30 mL of chloroform. Dilute the eluate with chloroform to volume. Spectrometric conditions Mode: UV Analytical wavelength: 306 nm Cell: 1 cm Blank: A solvent mixture of the same composition as that used for the Standard solution Analysis Samples: Standard solution and Sample solution Acceptance criteria: The absorbance of the Sample solution does not exceed that of the Standard solution (NMT 0.75%, calculated on the labeled aspirin content). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS
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hydrochloric acid or 2 N sodium hydroxide to a pH of 6.8 0.05. Standard solution: USP Aspirin RS of a known concentration in the same medium Sample solution: Filtered portion of the solution under test, diluted, if necessary, with 0.1 N hydrochloric acid (analyzing the Acid Stage) and with Diluent (analyzing the Buffer Stage) Analysis Samples: Standard solution and Sample solution Determine the amount of C9H8O4 dissolved by determining UV absorbances at the wavelength of the isosbestic point of aspirin and salicylic acid (280 nm in the Acid Stage, and 265 nm in the Buffer Stage) of the Sample solution in comparison to the Standard solution. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase and Diluent: Prepare as directed in the Assay. Standard solution: 0.015 mg/mL of salicylic acid from USP Salicylic Acid RS dissolved in the Standard solution from the Assay. Sample solution: Use the Sample stock solution from the Assay. Chromatographic system: Proceed as directed under Assay. System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and aspirin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin Relative standard deviation: NMT 4.0% of the salicylic acid peak responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6O3 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) = nominal concentration of aspirin in the Sample CU solution as determined in the Assay (mg/mL) Acceptance criteria: NMT 3.0% rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label indicates that the Capsules or the contents thereof are enteric coated. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
Aspirin Delayed-Release Capsules

(Comment on this Monograph)id=m6260=Aspirin DelayedRelease Capsules=A-Monos.pdf) DEFINITION Aspirin Delayed-Release Capsules contain NLT 93.0% and NMT 107.0% of the labeled amount of aspirin (C9H8O4). IDENTIFICATION A. PROCEDURE Sample: 100 mg of Capsule contents Analysis: Boil it with 10 mL of water for several min. Cool, and add 1 drop of ferric chloride TS. Acceptance criteria: A violet-red color is produced. B. INFRARED ABSORPTION 197K Sample: Shake a quantity of the contents of Capsules, equivalent to 500 mg of aspirin, with 10 mL of alcohol for several min. Centrifuge the mixture. Pour off the clear supernatant and evaporate it to dryness. Dry the residue in a vacuum at 60 for 1 h. ASSAY PROCEDURE Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate in a mixture of 850 mL of water and 150 mL of acetonitrile, and adjust with glacial acetic acid to a pH of 3.4. Diluent: Acetonitrile and formic acid (99:1) Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Sample stock solution: Remove, as completely as possible, the contents of NLT 20 Capsules. Mix the combined contents, and transfer powder, equivalent to 100 mg of aspirin, to a suitable container. Add 20.0 mL of Diluent and 10 glass beads. Shake vigorously for 10 min, and centrifuge. Sample solution: Dilute a volume of the Sample stock solution with nine volumes of Diluent. [NOTERetain the remaining portion of Sample stock solution for the test for Procedure: Limit of free salicylic acid.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Aspirin RS in the Standard solution (mg/mL) CU = nominal concentration of aspirin in the Sample solution (mg/mL) Acceptance criteria: 93.0%107.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711: Proceed as directed under Apparatus 1 and Apparatus 2, Delayed-Release Dosage Forms, Method B. Apparatus 1: 100 rpm Time: 90 min, for Buffer Stage Diluent: 0.1 N hydrochloric acid and 0.20 M tribasic sodium phosphate (3:1). Adjust, if necessary, with 2 N
Aspirin Suppositories
(Comment on this Monograph)id=m6280=Aspirin Suppositories=A-Monos.pdf) DEFINITION Aspirin Suppositories contain NLT 90.0% and NMT 110.0% of the labeled amount of aspirin (C9H8O4).
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IDENTIFICATION [NOTEThe Sample is prepared as follows.] Sample: Transfer a portion of the melted Suppositories obtained in the Assay, equivalent to 1 g of aspirin, to a 125mL conical flask. Add 20 mL of alcohol, and warm until completely disintegrated. Cool in an ice bath for 5 min, filter, and evaporate the filtrate to dryness: the residue meets the requirements of the following tests. A. Heat the residue with water for several min, cool, and add 1 or 2 drops of ferric chloride TS: a violet-red color is produced. B. INFRARED ABSORPTION 197K ASSAY PROCEDURE [NOTEIn this Assay, use chloroform that recently was saturated with water.] Chromatographic column: Uniformly pack a chromatographic tube, as described in Procedure: Limit of free salicylic acid, with a mixture of 3 g of chromatographic siliceous earth and 2 mL of sodium bicarbonate solution (1 in 12) prepared on the day of use. Standard stock solution: Transfer 50 mg of USP Aspirin RS to a 50-mL volumetric flask, add 0.5 mL of glacial acetic acid, and add chloroform to volume. Standard solution: 0.05 mg/mL USP Aspirin RS from Standard stock solution diluted with a solution of glacial acetic acid in chloroform (1 in 100) Sample solution: Tare a small dish and glass rod, place in the dish NLT 5 Suppositories, heat gently on a steam bath until melted, then stir, and cool while stirring. Transfer a portion of the mass, equivalent to 50 mg of aspirin, to a 50mL volumetric flask containing 1 mL of a solution of hydrochloric acid in methanol (1 in 50), add 40 mL of chloroform, and add chloroform to volume. Spectrometric conditions Mode: UV Analytical wavelength: 280 nm Cell: 1 cm Blank: Chloroform Analysis: Pipet 5 mL of the Sample solution into the column, wash with 5 mL of chloroform, then again with 25 mL of chloroform, and discard the washings. Without delay, elute into a 100-mL volumetric flask with 10 mL of a solution of glacial acetic acid in chloroform (1 in 10), and then with 85 mL of a solution of glacial acetic acid in chloroform (1 in 100), and dilute with the latter solvent to volume. Without delay, determine the absorbances of the eluted Sample solution and Standard solution. Calculate the percentage of C9H8O4 in the portion of Suppositories taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Aspirin RS in the Standard solution (g/mL) CU = nominal concentration of aspirin in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% AU AS CS IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Ferric chloride-urea reagent: To a mixture of 8 mL of ferric chloride solution (6 in 10) and 42 mL of 0.05 N hydrochloric acid, add 60 g of urea. Dissolve the urea by

swirling and without the aid of heat, and adjust the resulting solution, if necessary, by the addition of 6 N hydrochloric acid to a pH of 3.2. [NOTEPrepare on the day of use.] Chromatographic column: Insert a small pledget of glass wool above the stem constriction of a 20- 2.5-cm chromatographic tube, and uniformly pack with a mixture of 1 g of chromatographic siliceous earth and 0.5 mL of 5 M phosphoric acid. Directly above this layer, pack a similar mixture of 3 g of chromatographic siliceous earth, and 2 mL of Ferric chloride-urea reagent. Standard stock solution: Dissolve a suitable quantity of salicylic acid in chloroform to obtain a solution containing 150 g/mL of salicylic acid. Standard solution: Pipet 5 mL of the Standard stock solution into a 50-mL volumetric flask containing 10 mL of methanol, 0.1 mL of hydrochloric acid, and 10 mL of a solution of glacial acetic acid in ether (1 in 10). Add chloroform to volume. Sample solution: Tare a small dish and glass rod, place in the dish NLT 5 Suppositories, heat gently on a steam bath until melted, then stir, and cool while stirring. Transfer a portion of the mass, equivalent to 50 mg of aspirin (see Sample solution in the Assay), to a small beaker. Add 10 mL of chloroform, warm slightly, and stir until dissolved. With the aid of 5 mL of chloroform, transfer to the chromatographic adsorption column. Pass 50 mL of chloroform in several portions through the column, rinse the tip of the chromatographic tube with chloroform, and discard the eluate. If the purple zone reaches the bottom of the tube, discard the column, and repeat the test with a smaller quantity of melted Suppositories. Elute the adsorbed salicylic acid into a 100-mL volumetric flask containing 20 mL of methanol and 0.2 mL of hydrochloric acid by passing two 10-mL portions of a solution of glacial acetic acid in water-saturated ether (1 in 10), and then 30 mL of chloroform, through the column, and dilute the eluate with chloroform to volume. Spectrometric conditions Mode: UV Analytical wavelength: 306 nm Cell: 1 cm Blank: A solvent mixture of the same composition as the Standard solution Analysis Samples: Standard solution and Sample solution Concomitantly determine the absorbances. Acceptance criteria: The absorbance of the Sample solution is NMT 3.0% that of the Standard solution. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, in a cool place. USP REFERENCE STANDARDS 11 USP Aspirin RS
Aspirin Tablets
(Comment on this Monograph)id=m6290=Aspirin Tablets=AMonos.pdf) DEFINITION Aspirin Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aspirin (C9H8O4). Tablets of larger than 81mg size contain no sweeteners or other flavors. [NOTETablets that are enteric-coated meet the requirements for Aspirin Delayed-Release Tablets.]
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be used to bring the Reference Standard into solution before dilution with Medium.] Sample solution: Sample per Dissolution 711. Dilute with Medium, if necessary, and filter. Analysis Sample: Standard solution and Sample solution Determine the amount of C9H8O4 dissolved from UV absorbances at the wavelength of the isosbestic point of aspirin and salicylic acid at 265 2 nm of the Sample solution in comparison with the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C9H8O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase and Diluent: Prepare as directed in the Assay. Standard solution: 0.015 mg/mL of salicylic acid in the Standard solution prepared as directed in the Assay. Sample solution: Use the Sample stock solution from the Assay. Chromatographic system: Proceed as directed in the Assay. System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and aspirin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0, between salicylic acid and aspirin Relative standard deviation: NMT 4.0% for the salicylic acid peak responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of salicylic acid (C7H6O3) in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of the salicylic acid from the Sample solution = peak response of the salicylic acid from the rS Standard solution = concentration of USP Salicylic Acid RS in the CS Standard solution (mg/mL) CU = concentration of aspirin in the Sample solution as determined in the Assay (mg/mL) Acceptance criteria: NMT 0.3%. For Tablets that are coated, NMT 3.0%. rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Preserve flavored or sweetened Tablets of 81-mg size or smaller in containers holding NMT 36 Tablets each. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
IDENTIFICATION A. PROCEDURE Sample: 1 Tablet Analysis: Crush and boil it with 50 mL of water for 5 min, cool, and add 1 or 2 drops of ferric chloride TS. Acceptance criteria: A violet-red color is produced. B. INFRARED ABSORPTION 197K Sample: Shake a quantity of finely powdered Tablets, equivalent to 500 mg of aspirin, with 10 mL of alcohol for several min. Centrifuge the mixture. Pour off the clear supernatant, and evaporate it to dryness. Dry the residue in a vacuum at 60 for 1 h. ASSAY PROCEDURE Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate in a mixture of 850 mL of water and 150 mL of acetonitrile, and adjust with glacial acetic acid to a pH of 3.4. Diluent: Acetonitrile and formic acid (99:1) Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Sample stock solution: Transfer an equivalent to 100 mg of aspirin, from finely powdered Tablets (NLT 20), to a suitable container. Add 20.0 mL of Diluent and 10 beads. Shake vigorously for 10 min, and centrifuge. Sample solution: Dilute a volume of the Sample stock solution with 9 volumes of Diluent. [NOTERetain the remaining portion of Sample stock solution for the test for Procedure: Limit of free salicylic acid.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of the aspirin from the Sample solution = peak response of the aspirin from the Standard rS solution = concentration of USP Aspirin RS in the Standard CS solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION 711 0.05 M acetate buffer: Mix 2.99 g of sodium acetate trihydrate and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05. Medium: 0.05 M acetate buffer; 500 mL Apparatus 1: 50 rpm Time: 30 min Standard solution: USP Aspirin RS of a known concentration in Medium [NOTEPrepare the Standard solution at the time of use. An amount of alcohol not to exceed 1% of the total volume of the Standard solution may
Buffered Aspirin Tablets

(Comment on this Monograph)id=m6291=Buffered Aspirin Tablets=A-Monos.pdf) DEFINITION Buffered Aspirin Tablets contain Aspirin and suitable buffering agents. Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aspirin (C9H8O4).
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IDENTIFICATION A. PROCEDURE Sample: 1 Tablet Analysis: Crush and boil it with 50 mL of water for 5 min, cool, and add 1 or 2 drops of ferric chloride TS. Acceptance criteria: A violet-red color is produced. B. INFRARED ABSORPTION 197K Sample: Shake a quantity of finely powdered Tablets, equivalent to 500 mg of aspirin, with 10 mL of alcohol for several min. Centrifuge the mixture. Pour off the clear supernatant, and evaporate it to dryness. Dry the residue in a vacuum at 60 for 1 h. ASSAY PROCEDURE Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate in a mixture of 850 mL of water and 150 mL of acetonitrile, and adjust with glacial acetic acid to a pH of 3.4. Diluent: Acetonitrile and formic acid (99:1) Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Sample stock solution: Transfer an equivalent to 100 mg of aspirin, from finely powdered Tablets (NLT 20), to a suitable container. Add 20.0 mL of Diluent and 10 beads. Shake vigorously for 10 min, and centrifuge. Sample solution: Dilute a volume of the Sample stock solution with 9 volumes of Diluent. [NOTERetain the remaining portion of Sample stock solution for the test for Procedure: Limit of free salicylic acid.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of the aspirin from the Sample solution = peak response of the aspirin from the Standard rS solution = concentration of USP Aspirin RS in the Standard CS solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 0.05 M acetate buffer: Mix 2.99 g of sodium acetate trihydrate and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05. Medium: 0.05 M acetate buffer; 500 mL Apparatus 2: 75 rpm [NOTEWhere the Tablet is composed of multiple layers, a stainless steel wire helix may be used, if needed, to hold the Tablet in proper orientation in the apparatus.] Time: 30 min Standard solution: USP Aspirin RS in Medium [NOTE Prepare the Standard solution at the time of use. An amount of alcohol not to exceed 1% of the total volume of the rU

Standard solution may be used to bring the Reference Standard into solution prior to dilution with Medium.] Sample solution: Sample per Dissolution 711. Dilute with Medium, if necessary, and filter. Analysis Samples: Standard solution and Sample solution Determine the amount of C9H8O4 dissolved from UV absorbances at the wavelength of the isosbestic point of aspirin and salicylic acid at 265 2 nm of the Sample solution in comparison with a Standard solution having a known concentration. Tolerances: NLT 80% (Q) of the labeled amount of C9H8O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase and Diluent: Prepare as directed in the Assay. Standard solution: 0.015 mg/mL of salicylic acid in Diluent Sample solution: Use the Sample stock solution from the Assay. Chromatographic system: Proceed as directed in the Assay. System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and aspirin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin Relative standard deviation: NMT 4.0% for the salicylic acid peak responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6O3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of the salicylic acid from the Sample solution = peak response of the salicylic acid from the rS Standard solution = concentration of USP Salicylic Acid RS in the CS Standard solution (mg/mL) CU = concentration of aspirin in the Sample solution as determined in the Assay (mg/mL) Acceptance criteria: NMT 3.0% rU SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 1.9 mEq of acid is consumed for each 325 mg of aspirin in the Tablets. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
Aspirin Delayed-Release Tablets

(Comment on this Monograph)id=m6292=Aspirin DelayedRelease Tablets=A-Monos.pdf) DEFINITION Aspirin Delayed-Release Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of aspirin (C9H8O4).
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Analysis Samples: Standard solution and Sample solution Determine the quantity of C9H8O4 dissolved by determining UV absorbances at the wavelength of the isosbestic point of aspirin and salicylic acid (280 nm in the Acid stage, and 265 nm in the Buffer stage) of the Sample solution in comparison to the Standard solution. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase and Diluent: Prepare as directed in the Assay. Standard solution: 0.015 mg/mL of salicylic acid in Diluent Sample solution: Use the Sample stock solution from the Assay. Chromatographic system: Proceed as directed under the Assay. System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and aspirin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin Relative standard deviation: NMT 4.0% of the salicylic acid peak responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6O3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) = concentration of aspirin in the Sample solution as CU determined in the Assay (mg/mL) Acceptance criteria: NMT 3.0% rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label indicates that the Tablets are entericcoated. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
IDENTIFICATION A. PROCEDURE Sample: 1 Tablet Analysis: Crush and boil it with 50 mL of water for 5 min, cool, and add 1 or 2 drops of ferric chloride TS. Acceptance criteria: A violet-red color is produced. B. INFRARED ABSORPTION 197K Sample: Shake a quantity of finely powdered Tablets, equivalent to 500 mg of aspirin, with 10 mL of alcohol for several min. Centrifuge the mixture. Pour off the clear supernatant, and evaporate it to dryness. Dry the residue in a vacuum at 60 for 1 h. ASSAY PROCEDURE Mobile phase: 2 mg/mL of sodium 1-heptanesulfonate in acetonitrile and water (3:17), and adjust with glacial acetic acid to a pH of 3.4 Diluent: Acetonitrile and formic acid (99:1) Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Sample stock solution: Transfer an equivalent to 100 mg of aspirin, from finely powdered Tablets (NLT 20), to a suitable container. Add 20.0 mL of Diluent and 10 beads. Shake vigorously for 10 min, and centrifuge. Sample solution: Dilute a volume of the Sample stock solution with 9 volumes of Diluent. [NOTERetain the remaining portion of stock solution for the test for Procedure: Limit of free salicylic acid.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Aspirin RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 95.0%105.0% PERFORMANCE TESTS DISSOLUTION 711: Proceed as directed for Procedure for Method B under Procedure, Apparatus 1 and Apparatus 2, Delayed-Release Dosage Forms. Apparatus 1: 100 rpm Time: 90 min, for Buffer stage Diluent: 0.1 N hydrochloric acid and 0.20 M tribasic sodium phosphate (3:1), and adjust, if necessary, with 2 N hydrochloric acid or 2 N sodium hydroxide to a pH of 6.8 0.05 Standard solution: USP Aspirin RS of a known concentration in Medium Sample solution: Filtered portion of the solution under test, diluted, if necessary, with 0.1 N hydrochloric acid (analyzing the Acid stage) and with Diluent (analyzing the Buffer stage) rU rS CS
Aspirin Effervescent Tablets for Oral Solution

(Comment on this Monograph)id=m6293=Aspirin Effervescent Tablets for Oral Solution=A-Monos.pdf) DEFINITION Aspirin Effervescent Tablets for Oral Solution contain Aspirin and an effervescent mixture of a suitable organic acid and an alkali metal bicarbonate and/or carbonate. Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aspirin (C9H8O4).
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IDENTIFICATION A. PROCEDURE Analysis: Dissolve 1 Tablet in 50 mL of 1 N hydrochloric acid, boil for 5 min, and allow to cool. To 2 mL of the resulting solution add 2 or 3 drops of ferric chloride TS. Acceptance criteria: A violet-red color is produced. B. PROCEDURE Analysis: Add 1/2 of a Tablet to 50 mL of water in a flask, and immediately stopper with a stopper fitted with tubing so that the evolved gas passes through calcium hydroxide TS. Acceptance criteria: A white precipitate forms. ASSAY PROCEDURE Mobile phase: 2 mg/mL of sodium 1-heptanesulfonate in acetonitrile and water (3:17). Adjust with glacial acetic acid to a pH of 3.4. Diluent: Acetonitrile and formic acid (99:1) Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Sample stock solution: Transfer an equivalent to 100 mg of aspirin, from finely powdered Tablets (NLT 20), to a suitable container. Add 20.0 mL of Diluent and 10 beads. Shake vigorously for 10 min, and centrifuge. Sample solution: Quantitatively dilute a measured volume of the Sample stock solution with 9 volumes of Diluent. [NOTERetain the remaining portion of Sample stock solution for the test for Procedure: Limit of Free Salicylate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Aspirin RS in the Standard solution (mg/mL) CU = nominal concentration of aspirin in the Sample solution (tablet/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS SOLUTION TIME: 2 Tablets dissolve completely in 180 mL of water at 17.5 2.5 within 5 min. UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLATE Mobile phase and Diluent: Prepare as directed in the Assay. Standard solution: Dissolve a quantity of USP Salicylic Acid RS in the Standard solution, prepared as directed in the Assay, to obtain a solution having a known concentration of 0.015 mg/mL of salicylic acid. rU rS CS

Sample solution: Use the Sample stock solution prepared as directed in the Assay. Chromatographic system: Proceed as directed in the Assay. System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and aspirin are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin Relative standard deviation: NMT 4.0% of salicylic acid Analysis: Calculate the percentage of salicylic acid (C7H6O3) in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) = concentration of aspirin in the portion of tablets CU taken for the Sample solution, as determined in the Assay (mg/mL) Acceptance criteria: NMT 8.0% SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: consumed by 1 Tablet. NLT 5.0 mEq of acid is rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
Aspirin Extended-Release Tablets

(Comment on this Monograph)id=m6295=Aspirin ExtendedRelease Tablets=A-Monos.pdf) DEFINITION Aspirin Extended-Release Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of aspirin (C9H8O4). IDENTIFICATION A. PROCEDURE Sample: 1 Tablet Analysis: Crush and boil it with 50 mL of water for 5 min, cool, and add 1 or 2 drops of ferric chloride TS. Acceptance criteria: A violet-red color is produced. B. INFRARED ABSORPTION 197K Sample: Shake a quantity of finely powdered Tablets, equivalent to 500 mg of aspirin, with 10 mL of alcohol for several min. Centrifuge the mixture, pour off the clear supernatant, and evaporate it to dryness. Dry the residue in a vacuum at 60 for 1 h. ASSAY PROCEDURE Mobile phase: Dissolve 2 g of sodium 1-heptanesulfonate in a mixture of 850 mL of water and 150 mL of acetonitrile, and adjust with glacial acetic acid to a pH of 3.4. Diluent: Acetonitrile and formic acid (99:1) Standard solution: 0.5 mg/mL of USP Aspirin RS in Diluent Sample stock solution: Transfer an equivalent to 100 mg of aspirin, from finely powdered Tablets (NLT 20), to a suitable container. Add 20.0 mL of Diluent and 10 beads. Shake vigorously for 10 min, and centrifuge.
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Standard solution: USP Aspirin RS of a known concentration in Medium[NOTEPrepare the Standard solution at the time of use. An amount of alcohol not to exceed 5% of the total volume of the Standard solution may be used to bring the USP Reference Standard into solution prior to dilution with Medium.] Sample solutions: Filtered portions of the solution under test, suitably diluted with Medium, if necessary Analysis Samples: Standard solution and Sample solutions Determine the amount of C9H8O4 dissolved from UV absorbances at the wavelength of the isosbestic point of aspirin at about 265 nm of the Sample solutions in comparison with the Standard solution. Tolerances: The percentages of the labeled amount of C9H8O4 dissolved at the times specified conform to Acceptance Table 2.
Time (h) 1 2 4 8 Amount Dissolved (%) 1540 2560 3575 NLT 70
Sample solution: Dilute a volume of the Sample stock solution with 9 volumes of Diluent. [NOTERetain the remaining portion of Sample stock solution for the test for Procedure: Limit of Free Salicylic Acid.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of the aspirin from the Sample solution = peak response of the aspirin from the Standard rS solution = concentration of USP Aspirin RS in the Standard CS solution (mg/mL) = nominal concentration for the Sample solution CU (unit/mL) Acceptance criteria: 95.0%105.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1 [NOTEIf the product complies with this test, the labeling indicates that it meets USP Dissolution Test 1.] Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 60 rpm Time: 1 and 4 h Standard solution: USP Aspirin RS of a known concentration in Medium Sample solution: Filtered portions of the solution under test. Dilute with Medium, if necessary, and filter. Analysis Samples: Standard solution and Sample solution Determine the amount of C9H8O4 dissolved from UV absorbances at the wavelength of the isosbestic point of aspirin at 280 nm of the Sample solution in comparison with the Standard solution. Tolerances: The percentages of the labeled amount of C9H8O4 dissolved at the times specified conform to Acceptance Table 1.
Time (h) 1 4 Amount Dissolved (%) 2055 NLT 80
rU
IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase and Diluent: Prepare as directed in the Assay. Standard solution: 0.015 mg/mL of salicylic acid in the Standard solution prepared as directed in the Assay Sample solution: Use the Sample stock solution from the Assay. Chromatographic system: Proceed as directed in the Assay. System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and aspirin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin Relative standard deviation: NMT 4.0% of the salicylic acid peak responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6O3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of the salicylic acid from the Sample solution rS = peak response of the salicylic acid from the Standard solution CS = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) CU = concentration of aspirin in the Sample solution as determined in the Assay (mg/mL) Acceptance criteria: NMT 3.0% rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The labeling indicates the Dissolution Test with which the product complies.
Test 2 [NOTEIf the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2.] Medium: Water; 1000 mL Apparatus 2: 30 rpm Time: 1, 2, 4, and 8 h Detector: UV absorbances at the isosbestic point at 265 nm
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USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS

Mode: LC Detector: UV 280 nm Column: 4-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid, aspirin, phenacetin, and chloroform are about 0.3, 0.6, 1.0, and 1.8, respectively.] [NOTERecord each chromatogram until the chloroform peak.] Suitability requirements Resolution: NLT 2.0 between the salicylic acid, aspirin, and internal standard peaks Tailing factor: NMT 2.0 Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = ratio of the peak responses of aspirin and phenacetin from the Sample solution = ratio of the peak responses of aspirin and RS phenacetin from the Standard solution = concentration of USP Aspirin RS in the Standard CS solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% ALUMINUM HYDROXIDE Edetate disodium titrant: Dissolve 18.6 g of edetate disodium in water to make 1000 mL, and standardize the solution as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of edetate disodium titrant, and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: RU Result = W/26.98V = weight of aluminum in the portion of solution taken (mg) V = volume of edetate disodium titrant consumed (mL) Sample solution: To a portion of the powdered Tablets (NLT 20), equivalent to 250 mg of aluminum hydroxide, add 20 mL of water, stir, and slowly add 30 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, W
Aspirin, Alumina, and Magnesia Tablets

(Comment on this Monograph)id=m6300=Aspirin, Alumina, and Magnesia Tablets=A-Monos.pdf) DEFINITION Aspirin, Alumina, and Magnesia Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aspirin (C9H8O4), the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of aluminum hydroxide [Al(OH)3], and NLT 90.0% and NMT 110.0% of the labeled amount of magnesium hydroxide [Mg(OH)2]. IDENTIFICATION [NOTEThe Sample is prepared as follows.] Sample: To a 0.7-g portion of finely powdered Tablets, add 20 mL of 3 N hydrochloric acid and 5 drops of methyl red TS, heat to boiling, and add 6 N ammonium hydroxide until the color of the solution changes to deep yellow. Continue boiling for 2 min, and filter. The filtrate is used in Identification test B and the precipitate is used in Identification test C. A. The retention time of the aspirin peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Aspirin. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: The Sample filtrate is used. C. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the Sample precipitate with a hot solution of ammonium chloride (1 in 50), and dissolve the precipitate in hydrochloric acid. Acceptance criteria: The solution so obtained meets the requirements. ASSAY ASPIRIN Mobile phase: Mix 225 mg of tetramethylammonium hydroxide pentahydrate and 200 mg of sodium 1octanesulfonate in 700 mL of water. Add methanol, acetonitrile, and glacial acetic acid (150:150:1), and stir. Diluent: To 2 g of anhydrous citric acid, add 990 mL of acetonitrile, 990 mL of chloroform, and 20 mL of formic acid, and stir for 30 min. Allow to settle, and use the decanted clear solution. Internal standard solution: 2 mg/mL of phenacetin in Diluent Standard stock solution: 1 mg/mL of USP Salicylic Acid RS in Diluent Standard solution: Transfer about 325 mg of USP Aspirin RS to a 50-mL volumetric flask. Add 10.0 mL of Standard stock solution and 5.0 mL of Internal standard solution, dilute with Diluent to volume, and mix. Sample solution: Transfer a portion of the powdered Tablets (NLT 20), equivalent to about 325 mg of aspirin, to a screw-capped bottle, add 5.0 mL of Internal standard solution, and 45.0 mL of Diluent, cap the bottle, mix, and sonicate for 25 min. Centrifuge, and use the resultant clear solution. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 280 nm Column: 4-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid, aspirin, phenacetin, and chloroform are about 0.3, 0.6, 1.0, and 1.8, respectively.] [NOTERecord each chromatogram until the chloroform peak appears.] Suitability requirements Resolution: NLT 2.0 between the salicylic acid, aspirin, and internal standard peaks Tailing factor: NMT 2.0 Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of free salicylic acid in the Tablets taken: Result = (RU/RS) (CS/CU) 100 = ratio of the peak responses of salicylic acid and phenacetin from the Sample solution = ratio of the peak responses of salicylic acid and RS phenacetin from the Standard solution = concentration of USP Salicylic Acid RS in the CS Standard solution (mg/mL) = concentration of aspirin in the portion of Tablets CU taken for the Sample solution as determined in the Assay (mg/mL) Acceptance criteria: NMT 3.0% RU SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 1.9 mEq of acid is consumed for each 325 mg of aspirin in the Tablets. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
cool, and quantitatively transfer to a 200-mL volumetric flask with water and dilute with water to volume. Analysis: To 50 mL of Sample solution, add, in the order named and with continuous stirring, 25.0 mL of edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 50 mL of water for the Sample solution. Each mL of 0.05 M edetate disodium titrant consumed is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM HYDROXIDE Sample stock solution: Prepare as directed in the Assay for Aluminum hydroxide. Eriochrome black T indicator: Dissolve 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol. Analysis: To a volume of Sample solution, equivalent to 80 mg of magnesium hydroxide, add 200 mL of water, 20 mL of triethanolamine, 50 mL of ammoniaammonium chloride buffer TS, and 2 drops of Eriochrome black T indicator. Cool the solution to between 3 and 4 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS until the color changes to pure blue. Perform a blank determination, substituting water for the Sample solution, and make any necessary corrections. Each mL of 0.05 M edetate disodium is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g of sodium acetate (trihydrate), and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05; 900 mL Apparatus 2: 75 rpm Time: 45 min Detector: From UV absorbances at the wavelength of the isosbestic point of aspirin and salicylic acid at 265 2 nm Sample solutions: Filtered portions of the solution under test, suitably diluted with Medium Standard solution: USP Aspirin RS in Medium [NOTE Prepare the Standard solution at the time of use. An amount of methanol NMT 1% of the total volume of the Standard solution may be used to bring the Reference Standard into solution prior to dilution with Medium.] Tolerances: NLT 75% (Q) of the labeled amount of C9H8O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to aluminum hydroxide and to magnesium hydroxide, and for Content Uniformity with respect to aspirin. IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID [NOTEThe results from the Assay for Aspirin may be used for this test when calculated as described in the Analysis section of this test.] Mobile phase, Diluent, Internal standard solution, Standard stock solution, Standard solution, and Sample solution: Proceed as directed in Assay for Aspirin. Chromatographic system (See Chromatography 621, System Suitability.)
Aspirin, Alumina, and Magnesium Oxide Tablets

(Comment on this Monograph)id=m6302=Aspirin, Alumina, and Magnesium Oxide Tablets=A-Monos.pdf) DEFINITION Aspirin, Alumina, and Magnesium Oxide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of aspirin (C9H8O4), the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of aluminum hydroxide [Al(OH)3], and NLT 90.0% and NMT 110.0% of the labeled amount of magnesium oxide (MgO). IDENTIFICATION [NOTEThe Sample is prepared as follows.] Sample: To a 0.7-g portion of finely powdered Tablets, add 20 mL of 3 N hydrochloric acid and 5 drops of methyl red TS, heat to boiling, and add 6 N ammonium hydroxide until the color of the solution changes to deep yellow. Continue boiling
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for 2 min, and filter. The filtrate is used in Identification B and the precipitate is used in Identification C. A. The retention time of the aspirin peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Aspirin. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: The Sample filtrate is used C. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the Sample precipitate with a hot solution of ammonium chloride (1 in 50), and dissolve the precipitate in hydrochloric acid. The solution so obtained meets the requirements. D. PROCEDURE Analysis: Where the Tablets are composed of two layers, scrape a small amount of each layer into separate test tubes. Add 2 mL of water and 2 drops of methyl red TS to each tube, and shake for 15 s. Acceptance criteria: The solution from the aspirincontaining layer is red, and the solution from the buffercontaining layer is yellow. ASSAY ASPIRIN Mobile phase: Methanol, phosphoric acid, and water (30:3:70) Diluent: Hydrochloric acid and dehydrated alcohol (1:100) Aspirin standard stock solution: 5 mg/mL of USP Aspirin RS in Diluent, prepared by blending at high speed for 1.5 min Aspirin standard solution: 0.25 mg/mL of USP Aspirin RS from the Aspirin standard stock solution in dehydrated alcohol [NOTEUse these solutions within 1 h.] Salicylic acid standard stock solution: 5 mg/mL of USP Salicylic Acid RS in dehydrated alcohol. Transfer 3.0 mL of this solution to a 100-mL volumetric flask, and dilute with Diluent to volume. Salicylic acid standard solution: 7.5 g/mL of USP Salicyclic Acid RS from Salicylic acid standard stock solution in dehydrated alcohol System suitability solution: Transfer 5.0 mL of the Aspirin standard stock solution to a 100-mL volumetric flask, add 5.0 mL of the Salicylic acid standard stock solution, and dilute with dehydrated alcohol to volume. Sample solution: Transfer a counted number of Tablets, equivalent to 2500 mg of aspirin, to a 120-mL blender jar containing 100.0 mL of Diluent, and blend at high speed for 1.5 min. Immediately filter a portion of the mixture thus obtained, and transfer 1.0 mL of the filtrate to a 100-mL volumetric flask. Immediately dilute with dehydrated alcohol to volume. [NOTEPromptly inject this Sample solution into the chromatograph as directed for Analysis.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 205 nm Column: 4.6-mm 3-cm; 5-m packing L7 Flow rate: 3.5 mL/min Injection size: 10 L System suitability Samples: Aspirin standard solution, Salicylic acid standard solution, and System suitability solution [NOTEThe relative retention times for aspirin and salicylic acid are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the aspirin peak and the salicylic acid peak, System suitablity solution Tailing factor: NMT 2.0 for salicylic acid and aspirin peaks, Aspirin standard solution and Salicylic acid standard solution

Relative standard deviation: NMT 2.0% for salicylic acid and aspirin peaks, Aspirin standard solution and Salicylic acid standard solution Analysis Samples: Aspirin standard solution, Salicylic acid standard solution, and Sample solution Calculate the percentage of C9H8O4 in each Tablet taken: Result = (rU/rS) (CS/CU) 100 = aspirin peak responses from the Sample solution = aspirin peak responses from the Standard solution = concentration of USP Aspirin RS in the Aspirin standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% ALUMINUM HYDROXIDE Edetate disodium titrant: Dissolve 18.6 g of edetate disodium in water to make 1000 mL, and standardize the solution as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of edetate disodium titrant, and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: rU rS CS Result = W/Ar V = weight of aluminum in the portion of solution taken (mg) = atomic weight of aluminum, 26.98 Ar V = volume of Edetate disodium titrant consumed (mL) Sample solution: To a portion of the powdered Tablets (NLT 20), equivalent to 600 mg of aluminum hydroxide, add 20 mL of water, stir, and slowly add 30 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and transfer to a 200-mL volumetric flask. Wash the beaker with water, adding the washings to the flask, and add water to volume. Analysis: To 20 mL of the Sample solution, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acid-ammonium acetate buffer TS, and heat the solution near the boiling temperature for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from greenviolet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary corrections. Each mL of 0.05 to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM OXIDE Sample solution: Prepare as directed in the Assay for Aluminum hydroxide. Eriochrome black T indicator: Dissolve 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol. Analysis: To a volume of Sample solution equivalent to 40 mg of magnesium oxide add, mixing, 20 mL of W
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Mobile phase: Methanol, phosphoric acid, and water (30:3:70) Diluent: Hydrochloric acid and dehydrated alcohol (1:100) Aspirin standard stock solution: 5 mg/mL of USP Aspirin RS in Diluent, prepared by blending at high speed for 1.5 min Aspirin standard solution: 0.25 mg/mL of USP Aspirin RS from the Aspirin standard stock solution in dehydrated alcohol [NOTEUse these solutions within 1 h.] Salicylic acid standard stock solution: 5 mg/mL of USP Salicylic Acid RS in dehydrated alcohol. Transfer 3.0 mL of this solution to a 100-mL volumetric flask, and dilute with Diluent to volume. Salicylic acid standard solution: 7.5 g/mL of USP Salicylic Acid RS from Salicylic acid standard stock solution in dehydrated alcohol System suitability solution: Transfer 5.0 mL of the Aspirin standard stock solution to a 100-mL volumetric flask, add 5.0 mL of the Salicylic acid standard stock solution, and dilute with dehydrated alcohol to volume. Sample solution: Transfer a counted number of Tablets, equivalent to 2500 mg of aspirin, to a 120-mL blender jar containing 100.0 mL of Diluent, and blend at high speed for 1.5 min. Immediately filter a portion of the mixture thus obtained, and transfer 1.0 mL of the filtrate to a 100mL volumetric flask. Immediately dilute with dehydrated alcohol to volume.[NOTEPromptly inject this Sample solution into the chromatograph as directed for Analysis.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 205 nm Column: 4.6-mm 3-cm; 5-m packing L7 Flow rate: 3.5 mL/min Injection size: 10 L System suitability Samples: Aspirin standard solution, Salicylic acid standard solution, and System suitability solution [NOTEThe relative retention times for aspirin and salicylic acid are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the aspirin peak and the salicylic acid peak, System suitablity solution Tailing factor: NMT 2.0 for salicylic acid and aspirin peaks, Aspirin standard solution and Salicylic acid standard solution Relative standard deviation: NMT 2.0% for salicylic acid and aspirin peaks, Aspirin standard solution and Salicylic acid standard solution Analysis Samples: Aspirin standard solution, Salicylic acid standard solution, and Sample solution Calculate the percentage of free salicylic acid in the Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = salicylic acid peak responses from the Sample solution = peak responses from the Salicylic acid standard solution = concentration of USP Salicylic Acid RS in the Salicylic acid standard solution (g/mL) = nominal concentration of the Sample solution (mg/mL)
triethanolamine and 200 mL of water. Cool the solution for 10 min, while stirring, by immersion in an ice bath. Remove from the ice bath, and add 15 mL of ammoniaammonium chloride buffer TS and 2 drops of eriochrome black T indicator. Titrate with 0.05 M edetate disodium VS to a blue endpoint, allowing 60 s between drops of titrant as the endpoint is approached (after first color change is observed). [NOTEThe titration should be completed within 10 min after the addition of the buffer and indicator. If any precipitate is observed prior to titration, the solution should be discarded and a new solution prepared.] Perform a blank determination, substituting water for the Sample solution and make any necessary correction. Each mL of 0.05 M edetate disodium consumed is equivalent to 2.015 mg of MgO. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g of sodium acetate (trihydrate) and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05; 900 mL Apparatus 1 (10-mesh screen): 100 rpm Time: 45 min Detector: Determine the amount of C9H8O4 dissolved employing the following method. Alkaline detergent solution: 1 N sodium hydroxide and 30% solution of polyoxyethylene (23) lauryl ether (1000:0.5) pH 4.3 buffer detergent: 12.9 mg/mL of citric acid monohydrate and 20.6 mg/mL of dibasic sodium phosphate heptahydrate in water. Add 0.5 mL of a 30% solution of polyoxyethylene (23) lauryl ether. Standard solution: 0.45 mg/mL of USP Aspirin RS in Medium Analysis: Use an automatic analyzer consisting of (1) a liquid sampler, (2) a proportioning pump, (3) a suitable fluorometer equipped with a 0.4-cm flow cell and suitable recording devices, and (4) a manifold consisting of the components illustrated in the diagram in Automated Methods of Analysis 16. With the sample line pumping pH 4.3 buffer detergent, the other lines pumping their respective reagents, the fluorometer set at an excitation wavelength of 298 nm and an emission wavelength of 425 nm, adjust the system until a steady fluorescence baseline has been achieved. Start the sampler, and conduct determinations at a rate of 40/h, using a ratio of 5:1 for sample and wash time. Record the fluorescence values of the Standard solution and the solution under test. Calculate the percentage of C9H8O4 dissolved: Result = CS V (FU/FS) 100/L = concentration of USP Aspirin RS in the Standard solution (mg/mL) V = volume of medium (mL), 900 = fluorescence values of the solution under test FU = fluorescence values of the Standard solution FS L = Tablet label claim (mg) Tolerances: NLT 75% (Q) of the labeled amount of C9H8O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to aluminum hydroxide and to magnesium oxide, and for Content Uniformity with respect to aspirin IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID [NOTEThe results from the Assay for Aspirin may be used for this test when calculated as described in the Analysis section of this test.] CS
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System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 2.5 between the caffeine and dihydrocodeine peaks, NLT 1.0 between the dihydrocodeine and aspirin peaks, and NLT 1.5 between the aspirin and salicylic acid peaks, System suitability solution Relative standard deviation: NMT 2.0% for each analyte, Standard solution Analysis Samples: Standard solution, Salicylic acid standard solution, and Sample solution Calculate the percentage of C9H8O4, C8H10N4O2, and C18H23NO3 C4H6O6 in each Capsule taken: Result = (rU/rS) (CS/CU) 100 = peak response of the relevant analyte from the Sample solution = peak response of the relevant analyte from the rS Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of the appropriate analyte CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g of sodium acetate trihydrate and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05; 500 mL Apparatus 1: 50 rpm Time: 45 min Mobile phase and Chromatographic system: Prepare as directed in the Assay. Standard solution: Prepare a solution in Medium containing known concentrations of 0.002A mg of USP Aspirin RS, 0.002C mg of USP Caffeine RS, and 0.002D mg of USP Dihydrocodeine Bitartrate RS per mL, A, C, and D being the labeled amounts, in mg, of aspirin, caffeine, and dihydrocodeine bitartrate, respectively, in each Capsule. Sample solution: Filter a portion of the solution under test. Analysis: Separately inject equal volumes (10 L) of the Standard solution and the Sample solution into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the percentage of C9H8O4, C8H10N4O2, and C18H23NO3 C4H6O6 dissolved: Result = (rU/rS) CS V 100/L = peak response of the relevant analyte from the Sample solution = peak response of the relevant analyte from the rS Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) V = volume of the medium (mL), 500 L = Tablet label claim of the appropriate analyte (mg) Tolerances: NLT 75% (Q) of the labeled amount of C9H8O4, C8H10N4O2, and C18H23NO3 C4H6O6 is dissolved. rU
SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 1.9 mEq of acid is consumed for each 325 mg of aspirin in the Tablets. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Salicylic Acid RS
Aspirin, Caffeine, and Dihydrocodeine Bitartrate Capsules

(Comment on this Monograph)id=m6312=Aspirin, Caffeine, and Dihydrocodeine Bitartrate Capsules=A-Monos.pdf) DEFINITION Aspirin, Caffeine, and Dihydrocodeine Bitartrate Capsules contain NLT 90.0% and NMT 110.0% of the labeled amounts of aspirin (C9H8O4), caffeine (C8H10N4O2), and dihydrocodeine bitartrate (C18H23NO3 C4H6O6). IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution obtained as directed in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate and 2.3 g of monobasic ammonium phosphate in 850 mL of water. Add 150 mL of acetonitrile, degas, and adjust with phosphoric acid to a pH of 2.5. Diluent: Mix acetonitrile and water (46:53), and adjust with phosphoric acid to a pH of 2.5. Standard solution: 0001A mg/mL of USP Aspirin RS, 0.001C mg/mL of USP Caffeine RS, and 0.001D mg/mL of USP Dihydrocodeine Bitartrate RS in Diluent (A, C, and D being the labeled amounts, in mg, of aspirin, caffeine, and dihydrocodeine bitartrate, respectively, in each Capsule) [NOTEUse this solution within 3 h.] Salicylic acid standard solution: 0.005A mg/mL of USP Salicylic Acid RS in Diluent (A being the labeled amount, in mg, of aspirin per Capsule) [NOTEUse this solution within 3 h.] System suitability solution: 0.0001A mg/mL of USP Salicylic Acid RS (A being the labeled amount, in mg, of aspirin in each Capsule) [NOTEUse this solution within 3 h.] Sample solution: Transfer the contents of 10 Capsules to a 500-mL volumetric flask. Dilute with Diluent to volume. Transfer 5.0 mL of this mixture to a 100-mL volumetric flask, and dilute with Diluent to volume. Centrifuge a portion of this mixture, and use the clear supernatant as the Sample solution. [NOTEUse this solution within 3 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 215 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 2 mL/min Injection size: 10 L [NOTEThe relative retention times for caffeine, dihydrocodeine, aspirin, and salicylic acid are 0.2, 0.3, 0.7, and 1.0, respectively.]
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IDENTIFICATION PROCEDURE Diluent, Mobile phase, Sample solution, Chromatographic System, and System suitability: Proceed as directed in the Assay. Standard solution A: 3.3 mg/mL of USP Aspirin RS in Diluent Standard solution B: 1 mg/mL of USP Codeine Phosphate RS in Diluent Analysis Samples: Sample solution, Standard solution A, and Standard solution B Chromatograph these solutions as directed for the Assay. Acceptance criteria: The retention times of the major peaks of the Sample solution correspond to those of Standard solution A and Standard solution B. ASSAY PROCEDURE Mobile phase: Dissolve 225 mg of tetramethylammonium hydroxide pentahydrate and 200 mg of sodium 1octanesulfonate in 700 mL of water. Add 150 mL of methanol, 150 mL of acetonitrile, and 1.0 mL of glacial acetic acid, and stir. Diluent: To 15 g of anhydrous citric acid add 200 mL of methanol and 20 mL of glacial acetic acid, dilute with chloroform to 1000 mL, and mix until the citric acid is dissolved. Internal standard solution: 2 mg/mL of phenacetin in Diluent Salicylic acid standard stock solution: 1 mg/mL of USP Salicylic Acid RS in Diluent Salicylic acid standard solution: Salicylic acid standard stock solution, Internal standard solution, and Diluent (1:1:8) Codeine phosphate standard stock solution: Transfer 325J mg of USP Codeine Phosphate RS, to a 25-mL volumetric flask, J being the ratio of the labeled amount mg of codeine phosphate to the labeled amount mg of aspirin/Tablet. Dissolve in and dilute with Diluent to volume. Standard solution: 6.5 mg/mL of USP Aspirin RS in a solution of Codeine phosphate standard stock solution, Salicylic acid standard stock solution, Internal standard solution, and Diluent (5:1:1:3) Sample solution: Transfer an equivalent to 325 mg of aspirin, from finely powdered Tablets (NLT 20), to a screwcapped, 120-mL bottle, add 5.0 mL of Internal standard solution and 45.0 mL of Diluent, and sonicate for 25 min. Centrifuge, and use clear supernatant. [NOTEUse on the day prepared.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Samples: Salicylic acid standard solution and Standard solution [NOTEThe relative retention times for salicylic acid, aspirin, codeine, and phenacetin are about 0.3, 0.5, 0.8, and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin, between aspirin and codeine, and between codeine and phenacetin, Standard solution
IMPURITIES Organic Impurities PROCEDURE: LIMIT OF SALICYCLIC ACID [NOTEThe results from the Assay may be used for this test when calculated as described in the Analysis section of this test.] Mobile phase, Diluent, Standard solution, Salicylic acid standard solution, System suitability solution, and Sample solution: Proceed as directed in the Assay. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 215 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 2 mL/min Injection size: 10 L [NOTEThe relative retention times for caffeine, dihydrocodeine, aspirin, and salicylic acid are 0.2 , 0.3, 0.7, and 1.0, respectively.] System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 2.5 between the caffeine and dihydrocodeine peaks, NLT 1.0 between the dihydrocodeine and aspirin peaks, and NLT 1.5 between the aspirin and salicylic acid peaks, System suitability solution Relative standard deviation: NMT 2.0% for each analyte, Standard solution Analysis Samples: Standard solution, Salicylic acid standard solution, and Sample solution Calculate the percentage of salicylic acid in the Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response of salicylic acid from the Sample solution = peak response of salicylic acid from the Standard rS solution = concentration of USP Salicylic Acid RS in the CS Standard salicylic acid solution (mg/mL) = nominal concentration of aspirin in the portion CU of capsules taken of the Sample solution as determined in the Assay (mg/mL) Acceptance criteria: NMT 3.0% rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Caffeine RS USP Dihydrocodeine Bitartrate RS USP Salicylic Acid RS
Aspirin and Codeine Phosphate Tablets

(Comment on this Monograph)id=m6313=Aspirin and Codeine Phosphate Tablets=A-Monos.pdf) DEFINITION Aspirin and Codeine Phosphate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of aspirin (C9H8O4) and codeine phosphate hemihydrate (C18H21NO3 H3PO4 1/2H2O).
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Tailing factor: NMT 2.0 for each analyte peak Relative standard deviation: NMT 3.0% of the ratios of the peak responses of salicylic acid, aspirin, and codeine to the peak response of phenacetin, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 RU = peak response ratio of aspirin to phenacetin from the Sample solution = peak response ratio of aspirin to phenacetin RS from the Standard solution CS = concentration of USP Aspirin RS in the Standard solution (mg/mL) CU = nominal concentration for the Sample solution (mg aspirin/mL) Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in the portion of Tablets taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratio of codeine phosphate to phenacetin from the Sample solution = peak response ratio of codeine phosphate to RS phenacetin from the Standard solution = concentration of USP Codeine Phosphate RS in CS the Standard solution mg/mL = nominal concentration for the Sample solution CU (mg/mL) = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight of anhydrous codeine Mr2 phosphate, 397.37 Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS DISSOLUTION 711 0.05 M acetate buffer: Mix 2.99 g of sodium acetate trihydrate and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05. Medium: 0.05 M acetate buffer, 900 mL Apparatus 2: 75 rpm Time: 30 min Determine the amounts of C9H8O4 and C18H21NO3 H3PO4 1 /2H2O dissolved by employing the following method. Mobile phase and Diluent: Proceed as directed in the Assay. System suitability solution: Use the Standard solution from the Assay. Internal standard solution: 0.07 mg/mL of phenacetin in methanol Standard stock solution A: 0.36 mg/mL of USP Aspirin RS in Diluent Standard stock solution B: Transfer 12 mg of USP Codeine Phosphate RS and 25 mg of USP Salicylic Acid RS to a 50mL volumetric flask, and add 2.5 mL of methanol. Add Medium to volume. Pipet 10 mL of the resulting solution into a 100-mL volumetric flask, and add Medium to volume. Standard solutions A and B: Pipet 10 mL of Standard stock solution A and 10 mL of Standard stock solution B into separate containers, and add 3.0 mL of the Internal standard solution to each container. Sample solution: Withdraw a portion of the solution under test and filter, discarding the few mL of the filtrate. Pipet 10 mL of the filtrate and 3.0 mL of the Internal standard solution into a suitable container. Chromatographic system: Proceed as directed under Assay except to use the following injection size:

Injection size: 10 L for System suitability and 50 L for Analysis System suitability Sample: System suitability solution Proceed as directed in the Assay. Analysis Samples: Standard solution A, Standard solution B, and Sample solution Calculate the amount of codeine phosphate dissolved by comparison of the relative peak response ratios for the codeine phosphate peaks, obtained from Standard solution B and the Sample solution. Calculate the percentage of aspirin dissolved: Result = 100 [V C (RU/RS) + V C (RU/RS) (Mr1/Mr2)]/(L F) = volume of Medium (mL), 0.900 = concentration of USP Aspirin RS in Standard solution A (g/mL) = peak response ratio for the aspirin component RU from the Sample solution = peak response ratio for the aspirin component RS from the Standard solution A C = concentration of USP Salicylic Acid RS in Standard solution B (g/mL) RU = peak response ratio for the salicylic acid component from the Sample solution RS = peak response ratio for the salicylic acid component from the Standard solution B Mr1 = molecular weight of aspirin, 180.16 = molecular weight of salicylic acid, 138.12 Mr2 L = labeled amount of aspirin in 1 tablet (mg) F = conversion factor, 1000 g/mg Tolerances: NLT 75% (Q) of the labeled amounts of C9H8O4 and C18H21NO3 H3PO4 1/2H2O is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to aspirin and codeine phosphate. IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase, Diluent, Internal standard solution, Salicylic acid standard stock solution, Salicylic acid standard solution, Codeine phosphate standard stock solution, Standard solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Analysis Samples: Salicylic acid standard solution and Sample solution Calculate the percentage of free salicylic acid in the Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of salicylic acid to phenacetin from the Sample solution = peak response ratio of salicylic acid to RS phenacetin from the Salicylic acid standard solution = concentration of USP Salicylic Acid RS in the CS Salicylic acid standard solution (mg/mL) = nominal concentration of aspirin in the Sample CU solution Acceptance criteria: NMT 3.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. RU V C
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the labeled amount, in mg, of codeine phosphate to the labeled amount, in mg, of aspirin per Tablet.) Dissolve in and dilute with Diluent to volume. Standard solution: Transfer 65 mg of USP Aspirin RS to a 10-mL volumetric flask. Add 5.0 mL of Codeine phosphate standard stock solution, 1.0 mL of Salicylic acid standard stock solution, and 1.0 mL of Internal standard solution, and dilute with Diluent to volume. Sample solution: Transfer powdered Tablets (NLT 20), equivalent to 325 mg of aspirin, to a suitable vessel, add 5.0 mL of Internal standard solution and 45.0 mL of Diluent, and sonicate for 25 min. Centrifuge, and use the resultant clear solution. [NOTEUse on the day prepared.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Samples: Salicylic acid standard solution and Standard solution [NOTEThe relative retention times for salicylic acid, aspirin, codeine, and phenacetin are 0.3, 0.5, 0.8, and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin; NLT 2.0 between aspirin and codeine; NLT 2.0 between codeine and phenacetin, Standard solution Tailing factor: NMT 2.0 for each analyte peak, Salicylic acid standard solution and Standard solution Relative standard deviation: NMT 3.0% of the ratios of the peak responses of salicylic acid, aspirin, and codeine to the peak response of phenacetin, Salicylic acid standard solution and Standard solution Analysis Samples: Salicylic acid standard solution, Standard solution, and Sample solution Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of aspirin to phenacetin from the Sample solution RS = peak response ratio of aspirin to phenacetin from the Standard solution CS = concentration of USP Aspirin RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in the portion of Tablets taken: RU Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratio of codeine phosphate to phenacetin from the Sample solution RS = peak response ratio of codeine phosphate to phenacetin from the Standard solution CS = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Mr1 = molecular weight of codeine phosphate hemihydrate, 406.37 Mr2 = molecular weight of anhydrous codeine phosphate, 397.37 Acceptance criteria: 90.0%110.0% of the labeled amounts of C9H8O4 and C18H21NO3 H3PO4 1/2H2O. RU
USP REFERENCE STANDARDS 11 USP Aspirin RS USP Codeine Phosphate RS USP Salicylic Acid RS
Aspirin, Codeine Phosphate, Alumina, and Magnesia Tablets

(Comment on this Monograph)id=m6314=Aspirin, Codeine Phosphate, Alumina, and Magnesia Tablets=A-Monos.pdf) DEFINITION Aspirin, Codeine Phosphate, Alumina, and Magnesia Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of aspirin (C9H8O4), codeine phosphate hemihydrate (C18H21NO3 H3PO4 1/2H2O), aluminum hydroxide [Al(OH)3], and magnesium hydroxide [Mg(OH)2]. IDENTIFICATION PROCEDURE [NOTEThe Sample, Diluent, Aspirin standard solution, and Codeine phosphate standard solution are prepared as follows.] Sample: To a 0.7-g portion of finely powdered Tablets, add 10 mL of 3 N hydrochloric acid and 5 drops of methyl red TS, heat to boiling, and add 6 N ammonium hydroxide until the color of the solution changes to deep yellow. Continue boiling for 2 min, and filter. The filtrate is used in Identification B and the precipitate is used in Identification C. Diluent: To 15 g of anhydrous citric acid, add 200 mL of methanol and 20 mL of glacial acetic acid, dilute with chloroform to 1000 mL, and mix until the citric acid is dissolved. Aspirin standard solution: 3.3 mg/mL of USP Aspirin RS in the Diluent Codeine phosphate standard solution: 1 mg/mL of USP Codeine Phosphate RS in the Diluent A. The retention time of the aspirin and codeine peaks of the Sample solution, obtained as in the Assay for Aspirin and Codeine Phosphate, corresponds to that of the Aspirin standard solution and Codeine phosphate standard solution. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: The Sample filtrate is used. C. IDENTIFICATION TESTSGENERAL, Aluminum 191 Sample solution: Wash the Sample precipitate with a hot solution of ammonium chloride (1 in 50), and dissolve the precipitate in hydrochloric acid: the solution so obtained meets the requirements. ASSAY ASPIRIN AND CODEINE PHOSPHATE Mobile phase: Dissolve 225 mg of tetramethylammonium hydroxide pentahydrate and 200 mg of sodium 1octanesulfonate in 700 mL of water. Add 150 mL of methanol, 150 mL of acetonitrile, and 1.0 mL of glacial acetic acid, and stir. Diluent: To 15 g of anhydrous citric acid, add 200 mL of methanol and 20 mL of glacial acetic acid, dilute with chloroform to 1000 mL, and mix until the citric acid is dissolved. Internal standard solution: 2 mg/mL of phenacetin in Diluent Salicylic acid standard stock solution: 1 mg/mL of USP Salicylic Acid RS in Diluent Salicylic acid standard solution: Transfer 5.0 mL of Salicylic acid standard stock solution to a 50-mL volumetric flask, add 5.0 mL of Internal standard solution, and dilute with Diluent to volume. Codeine phosphate standard stock solution: 13J mg/mL of USP Codeine Phosphate RS in Diluent (J being the ratio of
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ALUMINUM HYDROXIDE Edetate disodium titrant: Dissolve 18.6 g of edetate disodium in water to make 1000 mL, and standardize the solution as follows. Weigh 2 g of aluminum wire, transfer to a 1000-mL volumetric flask, and add 50 mL of a mixture of hydrochloric acid and water (1:1). Swirl the flask to ensure contact of the aluminum and the acid, and allow the reaction to proceed until all of the aluminum has dissolved. Dilute with water to volume. Pipet 10 mL of this solution into a 250-mL beaker, add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS, and boil gently for 5 min. Cool, and add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS to a bright rose-pink color. Perform a blank determination, substituting 10 mL of water for the aluminum solution, and make any necessary correction. Calculate the molarity of the solution taken: Result = W/26.98V = weight of aluminum in the portion of solution taken (mg) V = volume of Edetate disodium titrant consumed (mL) Sample solution: Transfer an equivalent to 600 mg of aluminum hydroxide, from finely powdered Tablets (NLT 20), to a 150-mL beaker, add 20 mL of water, stir, and slowly add 30 mL of 3 N hydrochloric acid. Heat gently, if necessary, to aid solution, cool, and filter into a 200-mL volumetric flask. Wash the filter with water into the flask, and add water to volume. Analysis: Pipet 10 mL of Sample solution into a 250-mL beaker, add 20 mL of water, then add, in the order named and with continuous stirring, 25.0 mL of Edetate disodium titrant and 20 mL of acetic acidammonium acetate buffer TS. Add 50 mL of alcohol and 2 mL of dithizone TS. Titrate with 0.05 M zinc sulfate VS until the color changes from green-violet to rose-pink. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M Edetate disodium titrant is equivalent to 3.900 mg of Al(OH)3. Acceptance criteria: 90.0%110.0% MAGNESIUM HYDROXIDE Sample solution: Prepare as directed in the Assay for Aluminum hydroxide. Analysis: Pipet a volume of Sample solution, equivalent to 40 mg of magnesium hydroxide, into a 400-mL beaker, add 200 mL of water and 20 mL of triethanolamine, and stir. Add 10 mL of ammoniaammonium chloride buffer TS and 3 drops of an eriochrome black indicator solution (prepared by dissolving 200 mg of eriochrome black T in a mixture of 15 mL of triethanolamine and 5 mL of dehydrated alcohol, and mixing). Cool the solution to between 3 and 4 by immersion of the beaker in an ice bath, then remove, and titrate with 0.05 M edetate disodium VS to a blue endpoint. Perform a blank determination, substituting 10 mL of water for the Sample solution, and make any necessary correction. Each mL of 0.05 M edetate disodium consumed is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.05 M acetate buffer, prepared by mixing 2.99 g of sodium acetate trihydrate and 1.66 mL of glacial acetic acid with water to obtain 1000 mL of solution having a pH of 4.50 0.05; 900 mL Apparatus 2: 75 rpm Time: 30 min Determine the amounts of C9H8O4 and C18H21NO3 H3PO4 1 /2H2O dissolved by using the following method. W

Mobile phase, Diluent, and Aspirin and codeine phosphate standard solution: Prepare as directed in the Assay for Aspirin and codeine phosphate and Procedure: Limit of free salicylic acid. Internal standard solution: 0.07 mg/mL of phenacetin in methanol Standard stock solution A: 0.36 mg/mL of USP Aspirin RS in Diluent Standard solution A: To 10 mL of Standard stock solution A, add 3.0 mL of the Internal standard solution. Standard stock solution B: Transfer 12 mg of USP Codeine Phosphate RS and 25 mg of USP Salicylic Acid RS to a 50mL volumetric flask, and add 2.5 mL of methanol. Add Medium to volume. Pipet 10 mL of the resulting solution into a 100-mL volumetric flask, and add Medium to volume. Standard solution B: To 10 mL of Standard stock solution B, add 3.0 mL of the Internal standard solution. Sample solution: Withdraw a portion of the solution under test and filter, discarding the few mL of the filtrate. Pipet 10 mL of the filtrate and 3.0 mL of the Internal standard solution into a suitable container. Chromatographic system: Proceed as directed for Chromatographic system in the Assay for Aspirin and codeine phosphate except to use only the Standard solution for evaluation of the suitability of the system. Analysis: Proceed as directed in the Assay for Aspirin and codeine phosphate except to inject 50 L of Standard solution A, Standard solution B, and the Sample solution. Calculate the amount of codeine phosphate dissolved by comparison of the relative peak response ratios for the codeine phosphate peaks, of the Standard solution B, and the Sample solution. Calculate the percentage of aspirin dissolved: Result = [0.9 C (RU/RS) + 0.9C (RU/RS) (Mr1/Mr2)]/3.25 = concentration of USP Aspirin RS in Standard solution A (g/mL) = peak response ratio for the aspirin component RU from the Sample solution = peak response ratio for the aspirin component RS from Standard solution A C = concentration of USP Salicylic Acid RS in Standard solution B (g/mL) = peak response ratios for the salicylic acid RU component from the Sample solution = peak response ratio for the salicylic acid RS component from the Sample solution and Standard solution B = molecular weight of aspirin, 180.16 Mr1 = molecular weight of salicylic acid, 138.12 Mr2 Tolerances: NLT 75% (Q) of the labeled amounts of C9H8O4 and C18H21NO3 H3PO4 1/2H2O is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to aspirin and codeine phosphate and for Weight Variation with respect to aluminum hydroxide and magnesium hydroxide IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID [NOTEThe results from the Assay for Aspirin and codeine phosphate may be used for this test when calculated as described in the Analysis section of this test.] Mobile phase, Diluent, Salicylic acid standard stock solution, Salicylic acid standard solution, Codeine phosphate standard stock solution, Standard solution, and Sample solution: Proceed as directed in Assay for Aspirin and codeine phosphate. Internal standard solution: 2 mg/mL of phenacetin in Diluent C
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IDENTIFICATION INFRARED ABSORPTION 197K
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Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Samples: Salicylic acid standard solution and Standard solution [NOTEThe relative retention times for salicylic acid, aspirin, codeine, and phenacetin are 0.3, 0.5, 0.8, and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between salicylic acid and aspirin; NLT 2.0 between aspirin and codeine; NLT 2.0 between codeine and phenacetin Tailing factor: NMT 2.0 for each analyte peak Relative standard deviation: NMT 3.0% of the ratios of the peak responses of salicylic acid, aspirin, and codeine to the peak response of phenacetin Analysis: Calculate the percentage of free salicylic acid in the Tablets taken: Result = (RU/RS) (CS/CU) 100 RU = ratio of the peak response of salicylic acid to phenacetin from the Sample solution = ratio of the peak response of salicylic acid to RS phenacetin from the Salicylic acid standard solution = concentration of USP Salicylic Acid RS in the CS Salicylic acid standard solution (mg/mL) CU = nominal concentration of aspirin in the portion of tablets taken for the Sample solution as determined in the Assay for Aspirin and codeine phosphate (mg/mL) Acceptance criteria: NMT 3.0% NLT 1.9 mEq/Tablet
ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, diethylamine, and 0.13 M ammonium acetate (230:470:1.0:300). Adjust with glacial acetic acid to a pH of 7.5. Standard solution: 1 mg/mL of USP Astemizole RS in Mobile phase Sample solution: 1 mg/mL of Astemizole in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 4000 theoretical plates Tailing factor: NMT 1.8 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C28H31FN4O in the portion of Astemizole taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Astemizole RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Solution A: 17 mg/mL of tetrabutylammonium hydrogen sulfate in water Solution B: Acetonitrile Standard solution: 25 g/mL of USP Astemizole RS in methanol Sample solution: 10 mg/mL of Astemizole in methanol System suitability solution: 25 g/mL of USP Astemizole RS and 250 g/mL of ketoconazole in methanol Mobile phase: See the gradient table below.
Time (min) Equilibrate 5 0 15 Solution A (%) 0 95 95 80 80 0 0 Solution B (%) 100 5 5 20 20 100 100
SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Codeine Phosphate RS USP Salicylic Acid RS
Astemizole
(Comment on this Monograph)id=m6328=Astemizole=AMonos.pdf)
C28H31FN4O 458.58 1H-Benzimidazol-2-amine, 1-[(4-fluorophenyl)methyl]-N-[1-[2(4-methoxyphenyl)ethyl]-4-piperidinyl]-; 1-(p-Fluorobenzyl)-2-[[1-(p-methoxyphenethyl)-4piperidyl]amino]benzimidazole [68844-77-9]. DEFINITION Astemizole contains NLT 98.0% and NMT 102.0% of C28H31FN4O, calculated on the dried basis.
18 18.1 23
Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 278 nm Column: 4.6-mm 10-cm; contains base-deactivated 3m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 1.5 between the astemizole and ketoconazole peaks Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Astemizole taken: Result = (ri/rS) (CS/CU) 100 = peak response for each impurity = peak response from the Standard solution = concentration of USP Astemizole RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria Individual impurities: NMT 0.25% Total impurities: NMT 0.5% ri rS CS SPECIFIC TESTS LOSS ON DRYING 731: Dry it in a vacuum at 105 for 4 h: it loses NMT 0.5% of its weight. MELTING RANGE OR TEMPERATURE 741: 175178 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Astemizole RS
Official Monographs / Astemizole 297

ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, diethylamine and 0.13 M ammonium acetate (230:470:1.0:300). Adjust with glacial acetic acid to a pH of 7.5. Standard solution: 1 mg/mL of USP Astemizole RS in Mobile phase Sample solution: Equivalent to 1 mg/mL of astemizole, from powdered Tablets in Mobile phase [NOTEFinely powder NLT 20 Tablets. Mix a suitable quantity of powder with Mobile phase corresponding to 50% of the final volume for 30 min. Dilute to volume and centrifuge. Use the supernatant.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 4000 theoretical plates Tailing factor: NMT 1.8 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C28H31FN4O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Astemizole RS in the Standard solution (mg/mL) = nominal concentration of astemizole in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Simulated gastric fluid TS (without the enzyme); 800 mL Apparatus 2: 100 rpm Time: 45 min Detector: UV 285 nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Astemizole RS in Medium Tolerances: NLT 80% (Q) of the labeled amount of C28H31FN4O is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Mobile phase, Standard solution, and Chromatographic system: Proceed as directed in the Assay. Sample solution: Use the Sample solution from the Assay. Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Tablets taken: Result = (ri/rT) 100 ri rT = peak response for each impurity = sum of the responses of all of the peaks
Astemizole Tablets
(Comment on this Monograph)id=m6329=Astemizole Tablets=A-Monos.pdf) DEFINITION Astemizole Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of astemizole (C28H31FN4O). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY621 Standard solution: 1 mg/mL of USP Astemizole RS in methanol Sample solution: Equivalent to 1 mg/mL of Astemizole, from finely ground Tablets in methanol [NOTEFilter before use.] Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Toluene, dioxane, methanol, and ammonium hydroxide (60:30:10:1) Analysis Samples: Standard solution and Sample solution Develop the chromatogram in solvent system, until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, air-dry, and examine under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution.
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Astemizole / Official Monographs

rS CS
USP 32
= peak response from the Standard solution = concentration of USP Atenolol RS in the Standard solution (mg/mL) = concentration of Atenolol in the Sample solution CU (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% CHLORIDE AND SULFATE, Chloride 221 Sample solution: 10 mg/mL in 0.15 N nitric acid, made to 100 mL Acceptance criteria: Shows no more turbidity with 1 mL of silver nitrate TS than 1.4 mL of 0.020 N hydrochloric acid in 100 mL of 0.15 N nitric acid (0.1%) Organic Impurities PROCEDURE Mobile phase: Prepare as directed in the Assay. Sample solution 1: 0.1 mg/mL of Atenolol in Mobile phase Sample solution 2: 0.005 mg/mL Atenolol, made from Sample stock solution diluted with Mobile phase Chromatographic system: Proceed as directed in the Assay, except use the injection size listed below. Injection size: 50 L Analysis Samples: Sample solution 1 and Sample solution 2 [NOTEChromatograph Sample solution 1 for a period of time that is 6 times the retention time of the atenolol peak.] Calculate the percentage of each impurity observed in the chromatogram obtained from the Sample stock solution: Result = 0.5(ri/rA) = peak response of the Sample solution = response of the main atenolol peak of the Sample solution Acceptance criteria Individual impurities: NMT 0.25% Total impurities: NMT 0.5% ri rA SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 152156.5 LOSS ON DRYING 731: Dry it at 105 to constant weight: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at room temperature. USP REFERENCE STANDARDS 11 USP Atenolol RS
Acceptance criteria Individual impurities: NMT 0.25% Total impurities: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Astemizole RS
Atenolol
(Comment on this Monograph)id=m6330=Atenolol=AMonos.pdf)
266.34 C14H22N2O3 Benzeneacetamide, 4-[2-hydroxy-3-[(1methylethyl)amino]propoxy]-; 2-[p-[2-Hydroxy-3-(isopropylamino)propoxy]-phenyl]acetamide [29122-68-7]. DEFINITION Atenolol contains NLT 98.0% and NMT 102.0% of C14H22N2O3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 50 g/mL in methanol ASSAY PROCEDURE Mobile phase: 1.1 g of sodium 1-heptanesulfonate and 0.71 g of anhydrous dibasic sodium phosphate in 700 mL of water. Add 2 mL of dibutylamine, and adjust with 0.8 M phosphoric acid to a pH of 3.0. Add 300 mL of methanol, mix, and pass through a filter having a 0.5-m or finer porosity. Degas this solution before use. Standard solution: 0.01 mg/mL of USP Atenolol RS in Mobile phase Sample solution: 0.01 mg/mL of Atenolol in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 226 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 0.6 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H22N2O3 in the portion of Atenolol taken: Result = (rU/rS) (CS/CU) 100 rU = peak response from the Sample solution
Atenolol Injection
(Comment on this Monograph)id=m6332=Atenolol Injection=AMonos.pdf) DEFINITION Atenolol Injection is a sterile solution of Atenolol in Water for Injection. It contains a suitable buffering agent. It contains NLT 90.0% and NMT 110.0% of the labeled amount of atenolol (C14H22N2O3). IDENTIFICATION A. The retention time of the main peak of the Sample solution corresponds to that of the Standard solution, obtained in the Assay.
USP 32
B. ULTRAVIOLET ABSORPTION 197U Solution: 10 g/mL of atenolol in methanol ASSAY PROCEDURE Mobile phase: 930 mg of sodium octyl sulfate in 740 mL of water. Add 8 mL of 3.6 N sulfuric acid and pass through a 1-m or finer porosity filter. To the filtrate add 250 mL of acetonitrile. Citric acid buffer: 2.5 g of citric acid to a 500-mL volumetric flask. Add 400 mL of water, and swirl to dissolve. Adjust the solution with 2 N sodium hydroxide to a pH of 6.0, and dilute with water to volume. Standard stock solution: 50 mg of USP Atenolol RS to a 100-mL volumetric flask. Add 80 mL of Citric acid buffer, and sonicate for 30 s to achieve dissolution. Dilute with Citric acid buffer to volume. Standard solution: 0.2 mg/mL of USP Atenolol RS from Standard stock solution diluted with Citric acid buffer Sample solution: Nominally equivalent to 0.2 mg/mL of atenolol from Injection diluted with Citric acid buffer Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.7 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H22N2O3 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Atenolol RS in the Standard solution (mg/mL) CU = nominal concentration of atenolol in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 5.56.5 STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 33.3 USP Endotoxin Units/mg of atenolol. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, in a cool place or at a controlled room temperature, protected from light. Avoid freezing. USP REFERENCE STANDARDS 11 USP Atenolol RS USP Endotoxin RS rU rS CS
Official Monographs / Atenolol 299
Atenolol Oral Solution

(Comment on this Monograph)id=m386=Atenolol Oral Solution=A-Monos.pdf) DEFINITION Atenolol Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of C14H22N2O3. Prepare Atenolol Oral Solution at a 0.2% concentration, for example, as follows (see Pharmaceutical CompoundingNonsterile Preparations 795).
Atenolol Glycerin Vehicle for Oral Suspension Vehicle for Oral Solution, Sugar-Free To make 200 mg 5 mL 45 mL A sufficient quantity 100 mL
Calculate the quantity of each ingredient required for the total volume and atenolol strength to be prepared. Mix the Atenolol, previously pulverized, and Glycerin to form a smooth paste. Incorporate the Vehicle for Oral Suspension or an equal volume of Vehicle for Oral Solution, Sugar Free. [NOTEThe Vehicle for Oral Suspension may be omitted.] Incorporate sufficient Vehicle for Oral Solution, Sugar Free in increments to bring to volume, and mix well. [NOTEDo not use a sucrosecontaining vehicle for oral solution.] Package, and label. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Package in amber, tight containers, and store at controlled room temperature. LABELING: Label it to state that it is to be shaken well before use, and discarded after 60 days. Label it to state that it is to be kept out of reach of children. Label it to indicate the nominal atenolol concentration. BEYOND-USE DATE: NMT 60 days after preparation.
Atenolol Tablets
(Comment on this Monograph)id=m6335=Atenolol Tablets=AMonos.pdf) DEFINITION Atenolol Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of atenolol (C14H22N2O3). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Mix a quantity of powdered Tablets, equivalent to 100 mg of atenolol, with 15 mL of methanol, heat the mixture to 50, and shake for 5 min. Filter, and evaporate the filtrate on a water bath to dryness. Add 10 mL of 0.1 N hydrochloric acid to the residue, warm the solution, shake, and filter. To the filtrate add sufficient 1 N sodium hydroxide to make it alkaline, extract the solution with 10 mL of chloroform, drying the chloroform extract over anhydrous sodium sulfate. Filter the dried chloroform solution, evaporate the filtrate on a water bath to dryness, and dry the residue at 105 for 1 h. B. The retention time of the atenolol peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
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CS
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= concentration of USP Atenolol RS in the Standard solution (mg/mL) D = dilution factor for the Sample solution L = label claim of Tablet (mg atenolol) Tolerances: NLT 80% (Q) of the labeled amount of C14H22N2O3 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Atenolol RS
ASSAY PROCEDURE Mobile phase: 1.1 g of sodium 1-heptanesulfonate and 0.71 g of anhydrous dibasic sodium phosphate in 700 mL of water. Add 2 mL of dibutylamine, and adjust with 0.8 M phosphoric acid to a pH of 3.0. Add 300 mL of methanol and pass through a filter having a 0.5-m or finer porosity. Degas this solution before use. Standard solution: 0.01 mg/mL of USP Atenolol RS in Mobile phase Sample stock solution: Transfer 10 Tablets to a 1000-mL volumetric flask. Add 500 mL of Mobile phase, and sonicate for 15 min to disintegrate the Tablets. Dilute with Mobile phase to volume. Sample solution: Centrifuge a portion of the Sample stock solution, and dilute a volume of the supernatant with Mobile phase to obtain a solution nominally containing 0.01 mg/mL of atenolol. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 226 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 0.6 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H22N2O3 in each Tablet taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Atenolol RS in the Standard solution (mg/mL) = nominal concentration of atenolol in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N acetate buffer, pH 4.6, prepared by mixing 44.9 parts of 0.1 N sodium acetate with 55.1 parts of 0.1 N acetic acid solution; 900 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of C14H22N2O3 dissolved by using the following method. Mobile phase, Chromatographic system, and System suitability: Proceed as directed in the Assay. Standard solution: 0.01 mg/mL of USP Atenolol RS in Mobile phase Sample solution: Prepare a filtered portion of the solution under test. Quantitatively dilute an accurately measured volume of the filtrate with Mobile phase to obtain a solution estimated to contain 0.01 mg/mL of atenolol. Analysis: Proceed as directed in the Assay. Calculate the percentage of C14H22N2O3 dissolved: Result = V (rU/rS) CS D (100/L) V rU rS = volume of Medium, 900 mL = peak response from the Sample solution = peak response from the Standard solution
Atenolol and Chlorthalidone Tablets

(Comment on this Monograph)id=m6338=Atenolol and Chlorthalidone Tablets=A-Monos.pdf) DEFINITION Atenolol and Chlorthalidone Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of atenolol (C14H22N2O3) and chlorthalidone (C14H11ClN2O4S). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution A: 10 mg/mL of USP Chlorthalidone RS in methanol Standard solution B: 10J mg/mL of USP Atenolol RS in methanol, J being the ratio of the labeled amount, in mg, of atenolol to the labeled amount, in mg, of chlorthalidone/Tablet Sample solution: Equivalent to 10 mg/mL of chlorthalidone, from powdered Tablets in methanol. Shake a quantity of powdered Tablets in methanol for 15 min, and filter. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: n-Butyl alcohol and 1 N ammonium hydroxide (5:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Allow the spots to dry, and develop the chromatogram in a solvent system, until the solvent front has moved threefourths of the length of the plate. Locate the spots on the plate by viewing under short-wavelength UV light. Acceptance criteria: The principal spots of the Sample solution correspond in RF value, size, and intensity to those of the respective Standard solutions. B. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 250 mL of acetonitrile, 740 mL of water, and 8 mL of 3.6 N sulfuric acid. Add 930 mg of sodium octyl sulfate. Standard solution: Dissolve quantities of USP Atenolol RS and USP Chlorthalidone RS in a mixture of water and acetonitrile (3:1) to obtain a solution having concentrations of 0.25 mg of USP Chlorthalidone RS and 0.25J mg of USP Atenolol RS/mL, J being the ratio of the labeled amount, in mg, of atenolol to the labeled amount, in mg, of chlorthalidone/Tablet.
USP 32
Sample solution: Transfer 10 Tablets to a volumetric flask of such capacity that when filled to volume, a nominal concentration of 0.5 mg of chlorthalidone/mL is obtained. Add a mixture of water and acetonitrile (1:1) to half the capacity of the flask, and shake by mechanical means for NLT 15 min to disintegrate the Tablets. Dilute with a mixture of water and acetonitrile (1:1) to volume. Pass a portion of this stock solution through a filter having a 0.5m or finer porosity. Transfer 25.0 mL of the clear filtrate to a 50-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.7 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for atenolol and chlorthalidone are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the atenolol and chlorthalidone peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentages of C14H22N2O3 and C14H11ClN2O4S in each Tablet taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of the appropriate USP Reference Standard in the Standard solution (mg/mL) CU = nominal concentration of atenolol or chlorthalidone in the Sample solution (mg/mL) [NOTEIf a trailing peak or shoulder is observed on the chlorthalidone peak with a relative retention time of NMT 1.1 in the chromatograms of both the Standard solution and the Sample solution, sum the areas for the chlorthalidone peak with the trailing peak or shoulder to report the peak responses for chlorthalidone.] Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 45 min Determine the amounts of C14H22N2O3 and C14H11ClN2O4S dissolved by using the following method. Mobile phase, Chromatographic system, and System suitability: Proceed as directed in the Assay. Diluent: Acetonitrile and 3.6 N sulfuric acid (125:4) Standard solvent: Diluent and water (3:10) Standard solution: Dissolve USP Atenolol RS and USP Chlorthalidone RS in Standard solvent to obtain a solution having concentrations of 0.00085L mg of USP Atenolol RS and 0.00085L mg of USP Chlorthalidone RS/mL, L and L being the labeled amounts, in mg, of atenolol and chlorthalidone, respectively, per Tablet. Sample solution: Filtered solution under test and Diluent (10:3) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H22N2O3 and C14H11ClN2O4S dissolved: Result = V D (rU/rS) CS 100 rU rS CS V D rU rS CS = = = = =
Official Monographs / Atovaquone 301

volume of dissolution medium (mL), 900 dilution factor for the Sample solution peak response from the Sample solution peak response from the Standard solution nominal concentration of the appropriate USP Reference Standard in the Standard solution (mg/mL) Tolerances: NLT 80% (Q) of the labeled amount of C14H22N2O3, and NLT 70% (Q) of the labeled amount of C14H11ClN2O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity: Proceed as directed in the Assay, except prepare the Sample solution as follows. Transfer 1 Tablet to a volumetric flask of such capacity that when filled to volume, a nominal concentration of 0.25 mg of chlorthalidone/mL is obtained. Add a mixture of water and acetonitrile (1:1) to half the capacity of the flask, and shake by mechanical means for NLT 15 min to disintegrate the Tablet. Dilute with water to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity, and use the filtrate as the Sample solution. Calculate the percentage of C14H22N2O3 and C14H11ClN2O4S in the Tablet taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of the appropriate USP Reference Standard in the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Atenolol RS USP Chlorthalidone RS
Atovaquone
(Comment on this Monograph)id=m6342=Atovaquone=AMonos.pdf)
C22H19ClO3 366.84 1,4-Naphthalenedione, 2-[4-(4-chlorophenyl)cyclohexyl]-3hydroxy-, trans-; 2-[trans-4-(p-Chlorophenyl)cyclohexyl]-3-hydroxy-1,4naphthoquinone [95233-18-4]. DEFINITION Atovaquone contains NLT 97.5% and NMT 101.5% of C22H19ClO3, calculated on the anhydrous and organic solventfree basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
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USP 32
Analysis Samples: Standard solution, Sample solution, and Blank solution Transfer 12.0 mL of the Sample solution to a 50-mL colorcomparison tube, 10.0 mL of the Standard solution to another, and 10.0 mL of the Blank solution to a third. Then add 2.0 mL of the Sample solution to the Standard solution as well as the Blank solution. Add 2 mL of pH 3.5 Acetate Buffer (see Heavy Metals 231) to each of the three tubes. Add 1.2 mL of thioacetamide-glycerin base TS. Allow to stand for 2 min, and view downward over a white surface. Acceptance criteria: The solution from the Standard solution is slightly brown when compared with the solution from the Blank solution, and the color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (NMT 10 ppm). Organic Impurities PROCEDURE 1: RESIDUAL ORGANIC SOLVENTS Standard solution: 0.5 L of methanol, and 0.5 L of glacial acetic acid per mL of dimethylformamide Sample solution: 50 mg/mL of Atovaquone in dimethylformamide Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 2.8-m; contains 10% liquid phase G16 on S2 support Carrier gas: Nitrogen Temperature Column: 180 Detector: 250 Flow rate: 42.5 mL/min Injection size: 1 L System suitability Sample: Standard solution [NOTEThe relative retention times for methanol and acetic acid are about 0.4 and 1.0, respectively.] Suitability requirements Resolution: NLT 14 between methanol and acetic acid Column efficiency: NLT 700 for the acetic acid peak Tailing factor: NLT 0.8 for the acetic acid peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of methanol and acetic acid in the portion of Atovaquone taken: Result = (rU/rS) (CS/CU) F 100 = peak area of methanol or acetic acid from the Sample solution = peak area of methanol or acetic acid from the rS Standard solution = concentration of the Standard solution (mg/mL) CS = concentration of Atovaquone in the Sample CU solution (mg/mL) F = specific gravity (0.79 for methanol; 1.05 for glacial acetic acid) Acceptance criteria: NMT 0.2% methanol, NMT 0.2% acetic acid PROCEDURE 2 Analysis: Using the chromatograms of the Sample solution and the System suitability solution obtained in the Assay, calculate the percentage of atovaquone related compounds in the portion of Atovaquone taken: Result = (ri/rT) 100 ri = peak response of each impurity in the Sample solution rU
ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, water, and phosphoric acid (525:175:300:5) Diluent: Acetonitrile and water (4:1) Standard solution: 0.25 mg/mL of USP Atovaquone RS in Diluent System suitability solution: 0.25 mg/mL of USP Atovaquone RS and 0.02 mg/mL of USP Atovaquone Related Compound A RS in Diluent [NOTEStore in a low-actinic glass container.] Sample solution: 0.25 mg/mL of Atovaquone in Diluent [NOTEUse a low-actinic volumetric flask.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 3 mL/min Injection size: 20 L System suitability Sample: Standard solution and System suitability solution [NOTEThe relative retention times for atovaquone related compound A and atovaquone are about 0.85 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between atovaquone related compound A and atovaquone, System suitability solution Column efficiency: NLT 9000 theoretical plates, Standard solution Tailing factor: NMT 1.2, Standard solution Relative standard deviation: NMT 2%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H19ClO3 in the portion of Atovaquone taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Atovaquone RS in the Standard solution (mg/mL) = concentration of Atovaquone in the Sample CU solution (mg/mL) Acceptance criteria: 97.5%101.5% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS 231 Standard solution: Add 1.0 mL of Standard Lead Solution (see Heavy Metals 231, Special Reagents) to 0.5 g of magnesium oxide, and dry between 100 and 105. Proceed as directed for Sample solution, starting with Ignite to dull redness. Sample solution: Mix 1.0 g of Atovaquone with 0.5 g of magnesium oxide thoroughly in a silica crucible. Ignite to dull redness until a homogeneous white or grayish white mass is obtained. If the mixture remains colored after 30 min, allow to cool, mix using a fine glass rod, and repeat the ignition. If necessary, repeat the operation. Heat the residue at 800 for about 1 h. Cool, take up the residue in two 5-mL portions of 6 N hydrochloric acid, add 0.1 mL of phenolphthalein TS, and then add 13.5 N ammonium hydroxide until a pink color is obtained. Cool, add glacial acetic acid until the solution is decolorized, and add 0.5 mL in excess. Filter, if necessary, and wash the filter with water. Dilute with water to 20 mL. Blank solution: Proceed as directed for Sample solution, omitting the Atovaquone.
USP 32
rT = sum of all peak responses in the Sample solution Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 1.5 %
Impurity Table 1 Relative Retention Time 0.63 0.85 0.89 1.8 1.0 Acceptance Criteria, NMT(%) 0.5 1.0 0.3 0.5 0.2
Official Monographs / Atovaquone 303

with a mixture of methanol and water (1:1). [NOTE Minimize exposure of this solution to light.] Sample stock solution: Transfer a volume of the Oral Suspension equivalent to 5.2 g of the formulation to a lowactinic 250-mL volumetric flask. Add 50 mL of water, swirl for 5 min, add 150 mL of 0.1 M methanolic sodium hydroxide, and sonicate for 15 min. Allow to cool, and dilute with 0.1 M methanolic sodium hydroxide to volume. Immediately filter a 20-mL portion, discarding the first 5 mL of the filtrate. Sample solution: Transfer 3.0 mL of the clear filtrate of Sample stock solution to a low-actinic 100-mL volumetric flask, and dilute with a mixture of methanol and water (1:1) to volume. [NOTEMinimize exposure of this solution to light.] Chromatographic system (See Chromatography 621, System suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 12.5-cm; packing L1 Flow rate: 3 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for atovaquone related compound A and atovaquone are 0.86 and 1.0, respectively.] Suitability requirements Tailing factor: NMT 1, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H19ClO3 in each mL of the Oral Suspension taken: Result = (rU/rS) (CS/V) D (100/L) = atovaquone peak area from the Sample solution = atovaquone peak area from the Standard solution = concentration of USP Atovaquone RS in the Standard solution (mg/mL) V = volume of Oral Suspension taken to prepare the Sample solution (mL) D = dilution factor of the Sample solution L = label claim (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 For oral suspension packaged in single-unit containers: Meets the requirements DELIVERABLE VOLUME 698 For Oral Suspension packaged in multiple-unit containers: Meets the requirements IMPURITIES Organic Impurities PROCEDURE Analysis Samples: System suitability solution, Standard solution, and the Sample solution obtained in the Assay Using the chromatograms of the Samples, calculate the percentage of atovaquone-related compounds, based on the labeled strength of atovaquone: Result = (rU/rS) (CS/CU) (1/F) D (100/L)
Name Unnamed impurity Atovaquone related compound A Unnamed impurity Unnamed impurity Any other individual, unidentified impurity Sum of all other individual impurities
NMT 1.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Atovaquone RS USP Atovaquone Related Compound A RS
Atovaquone Oral Suspension

(Comment on this Monograph)id=m6344=Atovaquone Oral Suspension=A-Monos.pdf) DEFINITION Atovaquone Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H19ClO3. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Medium: Methanol and water (1:1) Sample solution: Dilute 5 mL of Sample solution from the Assay with Medium to 50 mL . Standard solution: Dilute 5 mL of Standard solution from Assay with Medium to 50 mL. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, phosphoric acid, and water (96:32:1:72) System suitability solution: 0.09 mg/mL of USP Atovaquone RS and 0.01 mg/mL of USP Atovaquone Related Compound A RS in 0.1 M methanolic sodium hydroxide [NOTEStore in a low-actinic glass container.] Standard stock solution: 30 mg of USP Atovaquone RS in a low-actinic 10-mL volumetric flask, and add 2 mL of water and 6 mL of 0.1 M methanolic sodium hydroxide. Sonicate for 5 min or until the material has dissolved. Allow to cool, and dilute with 0.1 M methanolic sodium hydroxide to volume. Standard solution: Transfer 3.0 mL of Standard stock solution to a low-actinic 100-mL volumetric flask, and dilute
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Atovaquone / Official Monographs

rU
USP 32
= individual peak response of an atovaquone related compound, if any, from the Sample solution rS = peak response of atovaquone from the Standard solution = concentration of USP Atovaquone RS in the CS Standard solution (mg/mL) = concentration of the Oral Suspension in the CU Sample solution (g/mL) F = relative response factor of an individual atovaquone related compound relative to the response of atovaquone: see Impurity Table 1 D = density of Oral Suspension (1.04 g/mL at 2025) L = labeled amount of atovaquone in the Oral Suspension (mg/mL) [NOTEDisregard any peak having a relative retention time of 0.3, which is due to photodegradation during preparation of the Sample solution.] Acceptance criteria: See Impurity Table 1
Impurity Table 1 Relative Retention Time 0.65 0.86 0.88 1.0 Relative Response Factor 1.08 0.85 1.0 1.0 1.0 Acceptance Criteria, NMT(%) 0.5 1.0 0.3 0.2 2.0
Atracurium Besylate
(Comment on this Monograph)id=m6350=Atracurium Besylate=A-Monos.pdf)
1243.48 C65H82N2O18S2 Isoquinolinium, 2,2-[1,5-pentanediylbis[oxy(3-oxo-3,1propanediyl)]]bis[1-[(3,4-dimethoxyphenyl)methyl]-1,2,3,4tetrahydro-6,7-dimethoxy-2-methyl-, dibenzenesulfonate; 2-(2-Carboxyethyl)-1,2,3,4-tetrahydro-6,7-dimethoxy-2methyl-1-veratrylisoquinolinium benzenesulfonate, pentamethylene ester [64228-81-5]. DEFINITION Atracurium Besylate contains NLT 96.0% and NMT 102.0% of C65H82N2O18S2, calculated on the anhydrous basis. It contains NLT 5.0% and NMT 6.5% of the trans-trans isomer, NLT 34.5% and NMT 38.5% of the cis-trans isomer, and NLT 55.0% and NMT 60.0% of the cis-cis isomer. IDENTIFICATION A. INFRARED ABSORPTION 197K: Meets the requirements B. The retention times of the three main isomeric peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer solution: 10.2 g of monobasic potassium phosphate in a 1000-mL volumetric flask. Dissolve in 950 mL of water. While stirring, adjust with phosphoric acid to a pH of 3.1, and dilute with water to volume. Solution A: Acetonitrile, methanol, and Buffer solution (4:1:15) Solution B: Acetonitrile, methanol, and Buffer solution (2:3:5) Mobile phase: See the gradient table below.
Name Atovaquone related compound Atovaquone related compound A Atovaquone related compound Atovaquone Any other atovaquone related compound Total related compound
SPECIFIC TESTS PH 791: 3.57.0 SEDIMENTATION For Oral Suspension packaged in multiple-unit containers Analysis: Transfer 50 mL of well-mixed Oral Suspension to a glass-stoppered graduated cylinder, and allow to stand for 16 h. Measure the volume, if any, of clear liquid observed in the cylinder. Acceptance criteria: NMT 1 mL of clear liquid ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Atovaquone RS USP Atovaquone Related Compound A RS
USP 32
Standard solution: 1 mg/mL of USP Atracurium Besylate RS in Solution A Sample solution: 1 mg/mL of Atracurium Besylate in Solution A Chromatographic system (See Chromatography, 621 System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; base-deactivated packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for the trans-trans isomer, the cis-trans isomer, and the cis-cis isomer are 0.8, 0.9, and 1.0 respectively.] Suitability requirements Resolution: NLT 1.1 between the trans-trans isomer and the cis-trans isomer and between the cis-trans isomer and the cis-cis isomer Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C65H82N2O18S2 in the portion taken: Result = (rT1/rT2) (CS/Cu) 100 = sum of the peak responses for the trans-trans isomer, the trans-cis isomer, and the cis-cis isomer from the Sample solution = sum of the peak responses for the trans-trans rT2 isomer, the trans-cis isomer, and the cis-cis isomer from the Standard solution CS = concentration of USP Atracurium Besylate RS in the Standard solution (mg/mL) = concentration of Atracurium Besylate in the CU Sample solution (mg/mL) Acceptance criteria: 96.0%102.0%, calculated on the anhydrous basis [NOTEIt contains 5.0%6.5% of the transtrans isomer, 34.5%38.5% of the cis-trans isomer, and 55.0%60.0% of the cis-cis isomer.] IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1 Buffer solution, Solution A, Solution B, and Mobile phase: Proceed as directed in the Assay. Standard solution: 1.0 mL of the Standard solution, prepared as directed in the Assay, diluted in Solution A to 100 mL Sample solution: Prepare as directed in the Assay Chromatographic system and System suitability: As directed in the Assay [NOTEFor identification purposes, the relative retention time for laudanosine is 0.3.] Analysis Samples: Standard solution and Sample solution Record the chromatograms, and measure all of the peak responses, except the three main isomeric peaks. Calculate the percentage of each impurity in the portion of C65H82N2O18S2 taken: Result = (ri/rS) (CS/CU) (1/F) 100 ri rS = peak response for each impurity from the Sample solution = cis-cis isomer peak response from the Standard solution rT1 CS
Official Monographs / Atracurium 305

= concentration of the cis-cis isomer in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU F = relative response factor of the impurity peak, which is 1.9 for laudanosine and 1.0 for all other unidentified impurities Acceptance criteria Laudanosine: NMT 0.5% Individual impurities: NMT 1.0% Total impurities: NMT 3.5% PROCEDURE 2: LIMIT OF METHYL BENZENESULFONATE Buffer solution, Solution A, Solution B: Prepare as directed in the Assay. Standard stock solution: 0.2 mg/mL of methyl benzenesulfonate in acetonitrile Standard solution: 1 g/mL of methyl benzenesulfonate from Standard stock solution diluted with Solution A Sample solution: 10 mg/mL of Atracurium Besylate in Solution A System suitability solution: Transfer 1 mL of the Sample solution and 5 mL of Standard stock solution to a 100-mL volumetric flask, and dilute with Solution A to volume. Mobile phase: See the gradient table below.
Time (min) 0 5 15 25 30 38 45 Solution A (%) 80 80 75 75 55 0 0 Solution B (%) 20 20 25 25 45 100 100
Chromatographic system Mode: LC Detector: UV 217 nm Column: 4.6-mm 25-cm; base-deactivated packing L1 Flow rate: 1 mL/min Injection size: 100 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 12.0 between the trans-trans isomer and methyl benzenesulfonate, System suitability solution Relative response: Responses for duplicate injections do not differ from each other by NMT 12% Analysis Samples: Standard solution and Sample solution Measure the responses for the methyl benzenesulfonate peaks. Acceptance criteria: NMT 0.01%, the peak response of the Sample solution being NMT that of the Standard solution PROCEDURE 3: LIMIT OF TOLUENE Standard solution: 100 g/mL of toluene in organic-free water (see Residual Solvents 467) Sample solution: 20 mg/mL of Atracurium Besylate in organic-free water (see Residual Solvents 467) Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.53-mm 30-m fused silica analytical column coated with a 5-m chemically cross-linked G27 stationary phase and a 0.53-mm 5-m silica guard column deactivated with phenylmethyl siloxane
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Atracurium / Official Monographs

Carrier gas: Helium with a linear velocity of 35 cm/s [NOTEWhen a makeup gas is used, nitrogen is recommended.] Temperature: See the temperature program table below.
Time (min) Injection port Detector port Column 0 5 22.5 24.9 40.9 Temperature 70 260 35 35 175 260 260
USP 32
Solution B: Acetonitrile, methanol, and Buffer solution (2:3:5) Mobile phase: See the gradient table below.
Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 15% of the toluene peak Analysis Samples: Standard solution and Sample solution Acceptance criteria: NMT 0.5% of toluene is found; the toluene peak from the Sample solution is NMT the toluene peak of the Standard solution. SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 5.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, in a cold place. [NOTEAtracurium Besylate is unstable at room temperature.] USP REFERENCE STANDARDS 11 USP Atracurium Besylate RS
Atracurium Besylate Injection

(Comment on this Monograph)id=m6356=Atracurium Besylate Injection=A-Monos.pdf) DEFINITION Atracurium Besylate Injection is a sterile solution containing NLT 90.0% and NMT 115.0% of the labeled amount of atracurium besylate (C65H82N2O18S2). It contains an amount of the transtrans-isomer equivalent to NLT 5.0% and NMT 6.5% of the labeled amount of atracurium besylate, an amount of the cistrans-isomer equivalent to NLT 34.5% and NMT 38.5% of the labeled amount of atracurium besylate, and an amount of the cis-cis-isomer equivalent to NLT 55.0% and NMT 60.0% of the labeled amount of atracurium besylate. [NOTEThe Injection is unstable at room temperature. Store all samples in the refrigerator. Analyze all preparations as soon as possible, or use a refrigerated injector.] IDENTIFICATION The retention times of the peaks of the three atracurium besylate isomers of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer solution: 10.2 g of monobasic potassium phosphate in a 1000-mL volumetric flask, and dissolve in 950 mL of water. While stirring, adjust with phosphoric acid to a pH of 3.1, and dilute with water to volume. Solution A: Acetonitrile, methanol, and Buffer solution (4:1:15)
Standard solution: 1 mg/mL of USP Atracurium Besylate RS in Solution A Sample solution: Nominally equivalent to 1 mg/mL of atracurium besylate from Injection diluted with Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; base-deactivated packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for atracurium besylate trans-trans-isomer, cis-trans-isomer, and cis-cis-isomer are about 0.8, 0.9, and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the atracurium besylate trans-trans-isomer and the cis-trans-isomer and between the atracurium besylate cis-trans-isomer and the cis-cisisomer Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Measure the responses for the three atracurium besylate isomer peaks. Calculate the percentage of C65H82N2O18S2 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = sum of the peak responses for the trans-trans isomer, the trans-cis isomer, and the cis-cis isomer from the Sample solution rS = sum of the peak responses for the trans-trans isomer, the trans-cis isomer, and the cis-cis isomer from the Standard solution = concentration of USP Atracurium Besylate RS in CS the Standard solution (mg/mL) = nominal concentration of atracurium besylate in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% of the labeled amount of C65H82 N2O18S2. It contains an amount of the trans-transisomer equivalent to 5.0%6.5% of the labeled amount of atracurium besylate, an amount of the cis-trans-isomer equivalent to 34.5%38.5% of the labeled amount of atracurium besylate, and an amount of the cis-cis-isomer equivalent to 55.0%60.0% of the labeled amount of atracurium besylate. rU IMPURITIES Organic Impurities PROCEDURE Buffer solution, Solution A, Solution B, Mobile phase, and Standard stock solution: Use the Standard solution prepared as directed in the Assay.
USP 32
System suitability solution: Heat a portion of the Standard stock solution at 90 for 30 min, and immediately chill to 5. Standard solution: 0.02 mg/mL from Standard stock solution diluted with Solution A Sample solution and Chromatographic system: Proceed as directed in the Assay. System suitability Sample: System suitability solution and Standard solution [NOTEThe retention times relative to the atracurium besylate cis-cis-isomer are 0.22 for the acidic compound; 0.29 for laudanosine; 0.44 and 0.50 for the trans- and cis-isomers, respectively, of the hydroxy compound; and 1.28 and 1.33 for the trans- and cis-isomers, respectively, of the monoacrylate.] Analysis Samples: Standard solution and Sample solution Measure the peak responses, except the peak due to benzenesulfonic acid occurring at a retention time of about 0.08 relative to the atracurium besylate cis-cisisomer. Calculate the percentage of each impurity in the portion of Sample solution taken: Result = (ri/rT) (CS/CU) 100 = peak response for each impurity from the Sample solution = sum of all the peak responses from the Standard rT solution = concentration of USP Atracurium Besylate RS in CS the Standard solution (mg/mL) = nominal concentration of atracurium besylate in CU the Sample solution (mg/mL) Acceptance criteria Acidic compound: NMT 6.0% Combined cis- and trans-isomers of the hydroxy compound: NMT 6.0% Laudanosine: NMT 3.0% Combined cis- and trans-isomers of the monoacrylate: NMT 3.0% Other known synthetic impurities: NMT 2.0% Other impurity: NMT 0.1 Total impurities: NMT 15.0% is found. ri SPECIFIC TESTS PH 791: 3.003.65 STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.56 USP Endotoxin Units/mg of atracurium besylate. INJECTIONS 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, in a refrigerator, and protect from freezing. Protect from light. USP REFERENCE STANDARDS 11 USP Atracurium Besylate RS
Official Monographs / Atropine 307
Atropine
(Comment on this Monograph)id=m6390=Atropine=AMonos.pdf)
289.37 C17H23NO3 Benzeneacetic acid, -(hydroxymethyl)-8-methyl-8-azabicyclo [3.2.1]oct-3-yl ester, endo-()-; 1H,5H-Tropan-3-ol ()-tropate (ester) [51-55-8]. DEFINITION Atropine contains NLT 99.0% and NMT 100.5% of C17H23NO3, calculated on the anhydrous basis. [CAUTIONHandle Atropine with exceptional care, since it is highly potent.] IDENTIFICATION A. PROCEDURE Standard solution: 36 mg of USP Atropine Sulfate RS Sample solution: 30 mg Analysis: Dissolve Standard solution and Sample solution in individual 60-mL separators with the aid of 5-mL portions of water. To each separator, add 1.5 mL of 1 N sodium hydroxide solution and 10 mL of chloroform. Shake for 1 min, allow the layers to separate, and pass the chloroform extracts through separate filters of 2 g of anhydrous granular sodium sulfate supported on pledgets of glass wool. Extract each aqueous layer with two additional 10-mL portions of chloroform, filtering and combining with the respective main extracts. Evaporate the chloroform solutions under reduced pressure to dryness, and dissolve each residue in 10 mL of carbon disulfide. Acceptance criteria: The IR absorption spectrum, determined in a 1-mm cell, of the solution of the Sample exhibits maxima only at the same wavelengths as that of the solution of the Standard. B. PROCEDURE Sample solution: A solution (1 in 50) in 3 N hydrochloric acid Analysis: Add gold chloride TS to the Sample solution. Acceptance criteria: A lusterless precipitate is formed (distinction from hyoscyamine, which, similarly treated, yields a lustrous precipitate). ASSAY PROCEDURE Sample: 400 mg of Atropine Analysis: Dissolve in 50 mL of glacial acetic acid. Titrate with 0.1 N perchloric acid VS to a green endpoint, using 1 drop of crystal violet TS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 28.94 mg of C17H23NO3.
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Atropine / Official Monographs

99.0%100.5%
USP 32
DEFINITION Atropine Sulfate contains NLT 98.5% and NMT 101.0% of (C17H23NO3)2 H2SO4, calculated on the anhydrous basis. [CAUTIONHandle Atropine Sulfate with exceptional care, because it is highly potent.] IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sulfate 191 Sample solution: 50 mg/mL Acceptance criteria: Meets the requirements ASSAY PROCEDURE Sample: 1 g Analysis: Dissolve in 50 mL of glacial acetic acid and titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 67.68 mg of (C17H23 NO3)2 H2SO4. Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities OTHER ALKALOIDS Sample: 150 mg Analysis: Dissolve in 10 mL of water. To 5 mL of the solution, add a few drops of platinic chloride TS: no precipitate is formed. To the remaining 5 mL of the solution, add 2 mL of 6 N ammonium hydroxide, and shake vigorously. Acceptance criteria: A slight opalescence may develop but no turbidity is produced. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class Ia 741: Not lower than 187, determined after drying at 120 for 4 h [NOTEBecause anhydrous Atropine Sulfate is hygroscopic, determine its melting temperature promptly on a specimen placed in the capillary tube immediately after drying.] OPTICAL ROTATION, Angular Rotation 781A: The observed rotation, in degrees, multiplied by 200, and divided by the length, in mm, of the polarimeter tube used, is between 0.60 and +0.05 (limit of hyoscyamine) Sample solution: 1 g, in water to make a volume of 20 mL at 25 ACIDITY Sample solution: 1.0 g Analysis: Dissolve in 20 mL of water. Titrate with 0.020 N sodium hydroxide. Acceptance criteria: NMT 0.30 mL is required to produce a yellow color. WATER DETERMINATION, Method I 921: NMT 4.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE 1: LIMIT OF FOREIGN ALKALOIDS AND OTHER IMPURITIES Standard solution: 24 mg/mL of USP Atropine Sulfate RS in methanol Sample solution A: 20 mg/mL of Atropine in methanol Sample solution B: 1 mg/mL of Atropine from Sample solution A diluted with methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.5-mm layer of chromatographic silica gel Application volume Standard solution: 5 L Sample solution A: 25 L Sample solution B: 1 L Developing solvent system: Chloroform, acetone, and diethylamine (5:4:1) Spray reagent: Potassium iodoplatinate TS Analysis: Proceed as directed under General Chapter. Allow the spots to dry, and develop the chromatogram in a solvent system, until the solvent front has moved threefourths of the length of the plate. Locate the spots on the plate by spraying with Spray reagent. Acceptance criteria: The RF value of the principal spot of each Sample solution corresponds to that of the Standard solution; no secondary spot of the Sample solution A exhibits intensity equal to or greater than the principal spot of the Sample solution B (NMT 0.2%). PROCEDURE 2: READILY CARBONIZABLE SUBSTANCES TEST 271 Sample solution: 200 mg in 5 mL of 2 N sulfuric acid Acceptance criteria: The solution has no more color than Matching Fluid A, and the solution is colored no more than light yellow upon the addition of 0.2 mL of nitric acid. SPECIFIC TESTS OPTICAL ROTATION, Angular Rotation 781A: 0.70 to +0.05 (limit of hyoscyamine) Sample solution: 1 g, previously dried at 105 for 1 h, in sufficient 50% alcohol (w/w) to obtain a volume of 20 mL at 25 (200-mm tube being used) MELTING RANGE OR TEMPERATURE 741: 114118 WATER DETERMINATION, Method I 921: NMT 0.2% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS
Atropine Sulfate
(Comment on this Monograph)id=m6400=Atropine Sulfate=AMonos.pdf) (C17H23NO3)2 H2SO4 H2O 694.83 676.83 (C17H23NO3)2 H2SO4 Benzeneacetic acid, -(hydroxymethyl)-, 8-methyl-8-azabicyclo [3.2.1]oct-3-yl ester, endo-()-, sulfate (2:1) (salt), monohydrate; 1H,5H-Tropan-3--ol ()-tropate (ester), sulfate (2:1) (salt) monohydrate [5908-99-6]. Anhydrous [55-48-1].
USP 32

Atropine Sulfate Injection

(Comment on this Monograph)id=m6410=Atropine Sulfate Injection=A-Monos.pdf) DEFINITION Atropine Sulfate Injection is a sterile solution of Atropine Sulfate in Water for Injection. It contains NLT 93.0% and NMT 107.0% of the labeled amount of (C17H23NO3)2 H2SO4 H2O. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: Chromatographic silica gel Sample solution: Use undiluted Application volume: 15 L Spray reagent: Potassium iodoplatinate TS Developing solvent system: Chloroform and diethylamine (9:1) Analysis: Proceed as directed under Thin-Layer Chromatographic Identification Test 201, the spots on the plate located by spraying with Spray reagent. Acceptance criteria: Meets the requirements ASSAY PROCEDURE Acetate buffer: 0.05 M sodium acetate, each L containing 2.9 mL of glacial acetic acid Mobile phase: 5.1 g of tetrabutylammonium hydrogen sulfate in a 1-L volumetric flask. Add 50 mL of acetonitrile, and dilute with Acetate buffer to volume. Adjust with 5 N sodium hydroxide to a pH of 5.5 0.1. Standard solution: 80 g/mL of USP Atropine Sulfate RS Sample solution: Nominally equivalent to 80 g/mL of atropine from Injection diluted with water System suitability solution: 2.5 g/mL of phydroxybenzoic acid. Dilute one volume of this solution with four volumes of the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 30-cm 3.9-mm; packing L1 Flow rate: 2 mL/min Injection size: 100 L System suitability Sample: Standard solution and System suitability solution [NOTEThe retention time of p-hydroxybenzoic acid is 1.6 relative to that of atropine.] Suitability requirements Resolution: NLT 2.2 between the p-hydroxybenzoic acid and atropine peaks, System suitability solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in each mL of the Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 Mr2 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Atropine Sulfate RS in the Standard solution (mg/mL) = nominal concentration of atropine in the Sample solution (mg/mL) = molecular weight of atropine sulfate monohydrate, 694.85 = molecular weight of anhydrous atropine sulfate, 676.83
SPECIFIC TESTS PH 791: 3.06.5 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 55.6 USP Endotoxin Units/mg of atropine sulfate. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS USP Endotoxin RS
Atropine Sulfate Ophthalmic Ointment

(Comment on this Monograph)id=m6420=Atropine Sulfate Ophthalmic Ointment=A-Monos.pdf) DEFINITION Atropine Sulfate Ophthalmic Ointment is Atropine Sulfate in a suitable ophthalmic ointment base. It contains NLT 90.0% and NMT 110.0% of the labeled amount of (C17H23NO3)2 H2SO4 H2O. It is sterile. IDENTIFICATION A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Sample solution: Transfer a portion of Ophthalmic Ointment, equivalent to 50 mg of atropine sulfate, to a suitable separator, and dissolve in 25 mL of ether. Add 25 mL of 0.01 N hydrochloric acid, shake vigorously, allow the layers to separate, and discard the organic phase. Heat the aqueous phase gently on a steam bath while passing nitrogen through the solution, to expel any residual ether. Analysis: Proceed as directed under IdentificationOrganic Nitrogenous Bases 181, beginning with In a second separator dissolve 50 mg. Acceptance critera: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Sulfate 191: Sample solution: Transfer 5 g of Ophthalmic Ointment to a separator, dissolve in 50 mL of ether, and extract with 20 mL of water. Acceptance critera: Meets the requirements ASSAY PROCEDURE pH 9.0 Buffer: 34.8 g of dibasic potassium phosphate in 900 mL of water. Adjust to a pH of 9.0 by the addition of 3 M hydrochloric acid or 1 M sodium hydroxide, as necessary, with mixing. Internal standard solution: 0.5 mg/mL of homatropine hydrobromide in water [NOTEPrepare fresh daily.] Standard solution: 0.1 mg/mL of USP Atropine Sulfate RS in water. Pipet 10 mL of this solution into a separator, add 2.0 mL of Internal standard solution and 5.0 mL of pH 9.0 Buffer, and adjust the solution in the separator with 1 M sodium hydroxide to a pH of 9.0. Extract with two 10-mL portions of methylene chloride, filter the methylene chloride extracts through 1 g of anhydrous sodium sulfate supported by a small cotton plug in a funnel into a 50-mL beaker, and evaporate under a stream of nitrogen to near-dryness. Dissolve the residue in 2.0 mL of methylene chloride. [NOTEPrepare fresh daily.] Sample solution: Transfer Ophthalmic Ointment, equivalent to 10 mg of atropine sulfate, to a separator containing 50 mL of ether, shake to dissolve, extract with three 25-mL portions of 0.1 M sulfuric acid, collect the acid extracts in a
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107.0% of the labeled amount of atropine sulfate [(C17H23NO3)2 H2SO4 H2O]. It may contain suitable stabilizers and antimicrobial agents. IDENTIFICATION A. INFRARED ABSORPTION 197M Sample: Ophthalmic Solution, equivalent to 30 mg, evaporated to dryness Standard: 36 mg USP Atropine Sulfate RS Analysis: Dissolve Sample and Standard in individual 60-mL separators with the aid of 5-mL portions of water. To each separator, add 1.5 mL of 1 N sodium hydroxide solution and 10 mL of chloroform. Shake for 1 min, allow the layers to separate, and filter the chloroform extracts through separate filters of 2 g of anhydrous granular sodium sulfate supported on pledgets of glass wool. Extract each aqueous layer with two additional 10-mL portions of chloroform, filtering and combining with the respective main extracts. Evaporate the chloroform solutions under reduced pressure to dryness, and dissolve each residue in 10 mL of carbon disulfide. Acceptance criteria: The IR absorption spectrum, determined in a 1-mm cell, of the solution of the Sample exhibits maxima only at the same wavelengths as that of the solution of the Standard. B. IDENTIFICATION TESTSGENERAL, Sulfate 191: Meets the requirements Sample solution: Evaporate to dryness a quantity of Ophthalmic Solution. Prepare a solution from the residue that contains the equivalent of 50 mg of atropine sulfate/mL. ASSAY PROCEDURE pH 9.0 buffer: 34.8 g of dibasic potassium phosphate in 900 mL of water. Adjust to a pH of 9.0, determined electrometrically, by the addition of 3 M hydrochloric acid or 1 M sodium hydroxide, as necessary, with mixing. Internal standard solution: 0.5 mg/mL of homatropine hydrobromide in water [NOTEPrepare fresh daily.] Standard solution: 0.1 mg/mL of USP Atropine Sulfate RS in water. [NOTEPrepare fresh daily.] Pipet 10 mL of this solution into a separator, add 2.0 mL of Internal standard solution and 5.0 mL of pH 9.0 buffer, and adjust the solution in the separator with 1 M sodium hydroxide to a pH of 9.0. Extract with two 10-mL portions of methylene chloride, filter the methylene chloride extracts through 1 g of anhydrous sodium sulfate supported by a small cotton plug in a funnel into a 50-mL beaker, and evaporate under a stream of nitrogen to near-dryness. Dissolve the residue in 2.0 mL of methylene chloride. Sample solution: Ophthalmic Solution, equivalent to 10 mg of atropine sulfate, in a 100-mL volumetric flask. Dilute with water to volume. Pipet 10 mL of this solution and treat as follows. Add 2.0 mL of Internal standard solution and 5.0 mL of pH 9.0 buffer, and adjust the solution in the separator with 1 M sodium hydroxide to a pH of 9.0. Extract with two 10-mL portions of methylene chloride, filter the methylene chloride extracts through 1 g of anhydrous sodium sulfate supported by a small cotton plug in a funnel into a 50-mL beaker, and evaporate under a stream of nitrogen to near-dryness. Dissolve the residue in 2.0 mL of methylene chloride. Chromatographic system Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m glass column packed with a 3% phase G3 on support S1AB
100-mL volumetric flask, dilute with 0.1 M sulfuric acid to volume. Pipet 10 mL of this solution and treat as follows. Add 2.0 mL of Internal standard solution and 5.0 mL of pH 9.0 Buffer, and adjust the solution in the separator with 1 M sodium hydroxide to a pH of 9.0. Extract with two 10-mL portions of methylene chloride, filter the methylene chloride extracts through 1 g of anhydrous sodium sulfate supported by a small cotton plug in a funnel into a 50-mL beaker, and evaporate under a stream of nitrogen to near-dryness. Dissolve the residue in 2.0 mL of methylene chloride. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m glass column packed with a 3% phase G3 on support S1AB Temperature: Column: 225 Injection port: 250 Detector: 250 Flow rate: 25 mL/min Carrier gas: Nitrogen Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 4.0 Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in the portion of Ophthalmic Ointment taken: Result = (RU/RS) (CS/CU) Mr1/Mr2 100 = peak area ratios of atropine sulfate to homatropine hydrobromide from the Sample solution = peak area ratios of atropine sulfate to RS homatropine hydrobromide from the Standard solution = concentration of USP Atropine Sulfate RS in the CS Standard solution (mg/mL) CU = concentration of the Sample solution (unit/mL) = molecular weight of atropine sulfate Mr1 monohydrate, 694.85 = molecular weight of anhydrous atropine sulfate, Mr2 676.83 Acceptance criteria: 90.0%110.0% SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements METAL PARTICLES IN OPHTHALMIC OINTMENTS 751: the requirements RU
Meets
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS
Atropine Sulfate Ophthalmic Solution

(Comment on this Monograph)id=m6430=Atropine Sulfate Ophthalmic Solution=A-Monos.pdf) DEFINITION Atropine Sulfate Ophthalmic Solution is a sterile, aqueous solution of Atropine Sulfate. It contains NLT 93.0% and NMT
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Temperature Column: 225 Injection port and detector: 250 Flow rate: 25 mL/min Carrier gas: Nitrogen Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 4.0 Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in each mL of Ophthalmic Solution taken: Result = (RU/RS) (CS/CU) Mr1/Mr2 100 = peak area ratios of atropine sulfate to homatropine hydrobromide from the Sample solution = peak area ratios of atropine sulfate to RS homatropine hydrobromide from the Standard solution = concentration of USP Atropine Sulfate RS in the CS Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of atropine sulfate Mr1 monohydrate, 694.85 = molecular weight of anhydrous atropine sulfate, Mr2 676.83 Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS PH 791: 3.56.0 STERILITY TESTS 71:

Adjust to a pH of 9.0, determined electrometrically, by the addition of 3 M hydrochloric acid or 1 M sodium hydroxide, as necessary, with mixing. Internal standard solution: 0.5 mg/mL of homatropine hydrobromide in water [NOTEPrepare fresh daily.] Standard solution: 0.1 mg/mL of USP Atropine Sulfate RS in water Pipet 10 mL of this solution into a separator, add 2.0 mL of Internal standard solution and 5.0 mL of pH 9.0 Buffer, and adjust the solution in the separator with 1 M sodium hydroxide to a pH of 9.0. Extract with two 10-mL portions of methylene chloride, filter the methylene chloride extracts through 1 g of anhydrous sodium sulfate supported by a small cotton plug in a funnel into a 50-mL beaker, and evaporate under a stream of nitrogen to near-dryness. Dissolve the residue in 2.0 mL of methylene chloride. [NOTEPrepare fresh daily.] Sample solution: Finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 1 mg of atropine sulfate, to a separator. Add 2.0 mL of Internal standard solution and 5.0 mL of pH 9.0 Buffer, and adjust the solution in the separator with 1 M sodium hydroxide to a pH of 9.0. Extract with two 10-mL portions of methylene chloride, filter the methylene chloride extracts through 1 g of anhydrous sodium sulfate supported by a small cotton plug in a funnel into a 50-mL beaker, and evaporate under a stream of nitrogen to near-dryness. Dissolve the residue in 2.0 mL of methylene chloride. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m glass column, packed with a 3% phase G3 on support S1AB Temperature Column: 225 Injection port and detector: 250 Flow rate: 25 mL/min Carrier gas: Nitrogen Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 4.0 Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of (C17H23NO3)2 H2SO4 H2O in the portion of Tablets taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 RU RS CS CU Mr1 Mr2 = peak area ratios of atropine sulfate to homatropine hydrobromide from the Sample solution = peak area ratios of atropine sulfate to homatropine hydrobromide from the Standard solution = concentration of USP Atropine Sulfate RS in the Standard solution (mg/mL) = nominal concentration of atropine sulfate in the Sample solution (mg/mL) = molecular weight of atropine sulfate monohydrate, 694.85 = molecular weight of anhydrous atropine sulfate, 676.83
Meets the requirements
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS
Atropine Sulfate Tablets

(Comment on this Monograph)id=m6440=Atropine Sulfate Tablets=A-Monos.pdf) DEFINITION Atropine Sulfate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of atropine sulfate [(C17H23NO3)2 H2SO4 H2O]. IDENTIFICATION A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181: Sample: A quantity of Tablets, equivalent to 5 mg of atropine sulfate Analysis: Triturate with 10 mL of water for a few minutes, and filter into a small separator. Render the solution alkaline with 6 N ammonium hydroxide, and extract with 50 mL of chloroform. Filter the chloroform layer, and evaporate to dryness. Acceptance criteria: The residue so obtained meets the requirements. B. IDENTIFICATION TESTSGENERAL, Sulfate 191: A filtered solution of Tablets meets the requirements of the tests. ASSAY PROCEDURE pH 9.0 Buffer: 34.8 g of dibasic potassium phosphate in 900 mL of water
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90.0%110.0%
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SPECIFIC TESTS LOSS ON DRYING 731: Dry at 105 to constant weight: it loses NMT 4.0% of its weight. POWDER FINENESS: Add 50 g to 450 mL of water containing 5 g of sodium pyrophosphate, and stir for 10 min. Pour the resulting dispersion slowly through a No. 325 standard sieve (see Particle Size Distribution Estimation by Analytical Sieving 786), and carefully wash the residue until clean. Dry the residue at 105 to constant weight: the dry weight of the residue so obtained is NMT 0.30% of the weight of the sample taken. Acceptance criteria: The dry weight of the residue is NMT 0.10% of the weight of the sample taken. MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the test for absence of Escherichia coli. PH 791: Disperse 1.0 g in 10 mL of carbon dioxide-free water: the pH of the mixed dispersion so obtained is 7.09.5. ADSORPTIVE CAPACITY: To 10 mL of a suspension (1 in 10) of the sample in water, add 80 mL of methylene blue solution (1 in 1000), and shake. Add 10 mL of barium chloride solution (1 in 50), and shake. Allow to stand for 15 min. Transfer 40 mL of the supernatant to a 50-mL centrifuge tube, and centrifuge. To 5 mL of the clear supernatant add 495 mL of water, and mix. Acceptance criteria: The color of the solution so obtained is not deeper than that of a solution containing 1.5 g of methylene blue per mL. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
PERFORMANCE TESTS DISINTEGRATION 701 Time: 15 min UNIFORMITY OF DOSAGE UNITS 905:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS
Activated Attapulgite
(Comment on this Monograph)id=m6470=Activated Attapulgite=A-Monos.pdf) DEFINITION Activated Attapulgite is a highly heat-treated, processed, native magnesium aluminum silicate. IDENTIFICATION PROCEDURE Analysis: Add 2 g in small portions to 100 mL of water, with vigorous agitation. Allow to stand for at least 12 h to ensure complete hydration. Place 2 mL of the resulting mixture on a suitable glass slide, and allow to air-dry at room temperature to produce a uniform film. Place the slide in a vacuum desiccator over a free surface of ethylene glycol. Evacuate the desiccator, and close the stopcock so that the ethylene glycol saturates the desiccator chamber. Allow to stand for 12 h. Record the X-Ray diffraction pattern (see X-Ray Diffraction 941), and calculate the d values. Acceptance criteria: Several peaks are observed; the characteristic peak corresponds to a d value between 10.3 and 10.7 Angstroms. IMPURITIES Inorganic Impurities LOSS ON IGNITION 733: When ignited at 1000 for 1 h, it loses 4.0%12.0% of its weight. ARSENIC AND LEAD: To 5.0 g add 50 mL of 1 N nitric acid, and boil for 30 min, adding 1 N nitric acid at times to maintain the volume. Filter into a 100-mL volumetric flask, wash the filter with water, and dilute the combined filtrate and washings with water to volume. Arsenic: Determine the arsenic in the solution by atomic absorption spectrometry (see Spectrophotometry and LightScattering 851), using a graphite furnace to volatilize the arsenic, as directed by the manufacturer of the instrument used, and measuring the absorbance at 189.0 nm against a standard. Acceptance criteria: NMT 2 ppm Lead: Determine the lead in the solution by atomic absorption spectrometry (see Spectrophotometry and LightScattering 851), using a graphite furnace to volatilize the lead, as directed by the manufacturer of the instrument used, and measuring the absorbance at 283.3 nm against a standard. Acceptance criteria: NMT 0.001% CARBONATE: Mix 1.0 g with 15 mL of 0.5 N sulfuric acid. Acceptance criteria: No effervescence occurs. ACID-SOLUBLE MATTER: Boil 2.0 g with 100 mL of 0.2 N hydrochloric acid for 5 min, and cool. Add water to adjust the volume to 100 mL, and filter. Evaporate 50 mL of the filtrate so obtained to dryness, and ignite the residue at 600. Acceptance criteria: NMT 0.25 g (25%). VOLATILE MATTER: Ignite at 600 for 1 h. Acceptance criteria: It loses 3.0%7.5% of its weight on the dried basis.
Colloidal Activated Attapulgite

(Comment on this Monograph)id=m6472=Colloidal Activated Attapulgite=A-Monos.pdf) DEFINITION Colloidal Activated Attapulgite is a purified native magnesium aluminum silicate. IDENTIFICATION PROCEDURE Analysis: Add 2 g in small portions to 100 mL of water, with vigorous agitation. Allow to stand for at least 12 h to ensure complete hydration. Place 2 mL of the resulting mixture on a suitable glass slide, and allow to air-dry at room temperature to produce a uniform film. Place the slide in a vacuum desiccator over a free surface of ethylene glycol. Evacuate the desiccator, and close the stopcock so that the ethylene glycol saturates the desiccator chamber. Allow to stand for 12 h. Record the X-Ray diffraction pattern (see X-Ray Diffraction 941), and calculate the d values: several peaks are observed. Acceptance criteria: The characteristic peak corresponds to a d value between 10.3 and 10.7 Angstroms. IMPURITIES Inorganic Impurities LOSS ON IGNITION 733: When ignited at 1000 for 1 h, it loses 17.0%27.0% of its weight. ARSENIC AND LEAD: To 5.0 g, add 50 mL of 1 N nitric acid, and boil for 30 min, adding 1 N nitric acid at times to maintain the volume. Filter into a 100-mL volumetric flask, wash the filter with water, and dilute the combined filtrate and washings with water to volume. Arsenic: Determine the arsenic in the solution by atomic absorption spectrometry (see Spectrophotometry and LightScattering 851), using a graphite furnace to volatilize the
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arsenic, as directed by the manufacturer of the instrument used, and measuring the absorbance at 189.0 nm against a standard. Acceptance criteria: NMT 2 ppm Lead: Determine the lead in the solution by atomic absorption spectrometry (see Spectrophotometry and LightScattering 851), using a graphite furnace to volatilize the lead, as directed by the manufacturer of the instrument used, and measuring the absorbance at 283.3 nm against a standard. Acceptance criteria: NMT 0.001% ACID-SOLUBLE MATTER: Boil 2.0 g with 100 mL of 0.2 N hydrochloric acid for 5 min, and cool. Add water to adjust the volume to 100 mL, and filter. Evaporate 50 mL of the filtrate so obtained to dryness, and ignite the residue at 600. Acceptance criteria: NMT 0.15 g (15%) CARBONATE: Mix 1.0 g with 15 mL of 0.5 N sulfuric acid. Acceptance criteria: No effervescence occurs. VOLATILE MATTER: When ignited at 600 for 1 h, it loses 7.5%12.5% of its weight on the dried basis. SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the test for absence of Escherichia coli. PH 791: Disperse 1.0 g in 10 mL of carbon dioxide-free water: the pH of the mixed dispersion so obtained is 7.09.5. LOSS ON DRYING 731: Dry at 105 to constant weight: it loses 5.0%17.0% of its weight. POWDER FINENESS: Add 50 g to 450 mL of water containing 5 g of sodium pyrophosphate, and stir for 10 min. Pour the resulting dispersion slowly through a No. 325 standard sieve (see Particle Size Distribution Estimation by Analytical Sieving 786), and carefully wash the residue until clean. Dry the residue at 105 to constant weight. Acceptance criteria: The dry weight of the residue so obtained is NMT 0.30% of the weight of the sample taken. ADSORPTIVE CAPACITY: To 10 mL of a suspension (1 in 10) of the specimen in water, add 80 mL of methylene blue solution (1 in 1000), and shake. Add 10 mL of barium chloride solution (1 in 50), and shake. Allow to stand for 15 min. Transfer 40 mL of the supernatant to a 50-mL centrifuge tube, and centrifuge. To 5 mL of the clear supernatant, add 495 mL of water, and mix. Acceptance criteria: The color of the solution so obtained is not deeper than that of a solution containing 1.5 g of methylene blue per mL. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Official Monographs / Aurothioglucose 313

DEFINITION Aurothioglucose contains NLT 95.0% and NMT 105.0% of C6H11AuO5S, calculated on the dried basis. It is stabilized by the addition of a small amount of Sodium Acetate. IDENTIFICATION A. PROCEDURE Standard solution: 4 mg/mL of USP Aurothioglucose RS Sample solution: 4 mg/mL Application volume: 10 L Developing solvent system: n-propyl alcohol, ethyl acetate, and water (3:1:3) Analysis Samples: Sample solution and Standard solution Apply Samples to a suitable thin-layer chromatographic glass microfilament sheet (see Chromatography 621) impregnated with silicic acid and a suitable fluorescing substance. Allow the spots to dry, and develop the chromatogram in a solvent system, until the solvent front has moved three-fourths of the length of the plate. Remove the sheet from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by examination under shortwavelength UV light. Acceptance criteria: The RF value of the principal spot obtained from the solution under test corresponds to that obtained from the Standard solution. B. PROCEDURE Analysis: To a portion of the filtrate obtained in the Assay, add barium chloride TS. Acceptance criteria: A heavy, white precipitate is formed. ASSAY PROCEDURE Analysis Sample: 1 g of Aurothioglucose Dissolve the Sample in 100 mL of water in a 300-mL Kjeldahl flask. Slowly add 10 mL of nitric acid, and when the reaction has subsided, boil the mixture for 5 min. Filter, wash well the separated gold with hot water, dry, and ignite to constant weight. The weight of the gold so obtained, multiplied by 1.991, represents the weight of C6H11AuO5S in the portion of Aurothioglucose taken. Acceptance criteria: 95.0%105.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +65 to +75 Sample solution: 10 mg/mL LOSS ON DRYING 731: Dry over phosphorus pentoxide for 24 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at room temperature. USP REFERENCE STANDARDS 11 USP Aurothioglucose RS
Aurothioglucose
(Comment on this Monograph)id=m6520=Aurothioglucose=AMonos.pdf)
Aurothioglucose Injectable Suspension

(Comment on this Monograph)id=m6545=Aurothioglucose Injectable Suspension=A-Monos.pdf) DEFINITION Aurothioglucose Injectable Suspension is a sterile suspension of Aurothioglucose in a suitable vegetable oil. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C6H11AuO5S. It may contain suitable thickening agents.
C6H11AuO5S Gold, (1-thio-D-glucopyranosato)-; (1-Thio-D-glucopyranosato)gold [12192-57-3].
392.18
314
Aurothioglucose / Official Monographs
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IDENTIFICATION PROCEDURE Standard solution: 4 mg/mL of USP Aurothioglucose RS Sample solution: Injectable Suspension, equivalent to 200 mg of aurothioglucose, in a centrifuge separator containing 20 mL of ethyl acetate and 50 mL of water Shake the mixture thoroughly, and centrifuge until the liquid phases have been clearly separated. Withdraw the lower, aqueous phase, and filter, discarding the first 10 mL of the filtrate. Collect the filtrate in a glass-stoppered vessel. Application volume: 10 L Developing solvent system: n-propyl alcohol, ethyl acetate, and water (3:1:3) Analysis Samples: Sample solution and Standard solution Apply the Sample to a suitable thin-layer chromatographic glass microfilament sheet (see Chromatography 621) impregnated with silicic acid and a suitable fluorescing substance. Allow the spots to dry, and develop the chromatogram in a solvent system, until the solvent front has moved about three-fourths of the length of the plate. Remove the sheet from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by examination under shortwavelength UV light. Acceptance criteria: The RF value of the principal spot from the solution under test corresponds to that from the Standard solution. ASSAY PROCEDURE Sample solution: Transfer with a pipet, calibrated to contain rather than to deliver, a measured volume of Injectable Suspension, equivalent to 200 mg of aurothioglucose, to a beaker containing 400 mL of acetone. Analysis: Wash the pipet into the beaker with a small quantity of acetone, allow the solids to settle, and decant the supernatant through a filter. Wash the solids with another 400-mL portion of acetone, and repeat the decantation. Transfer the solids to the filter with the aid of acetone, then transfer the filter and its contents to a shortnecked, 300-mL Kjeldahl flask, and add 5 mL of water and 20 mL of nitric acid. To this solution, add 15 mL of sulfuric acid slowly. Heat over a low flame, gently at first, and then increase the heat until fumes of sulfur trioxide are evolved. Allow the flask and contents to cool to room temperature, add 30 mL of water slowly, and 20 mL of hydrogen peroxide TS, again heat to fumes of sulfur trioxide, cool, and dilute with 30 mL of water. Pass the mixture through an ignited, tared filtering crucible, wash with water, heat the crucible and contents over a low flame to dry the precipitate, and ignite at 650 50 to constant weight. The weight of gold so obtained, multiplied by 1.991, represents the weight of C6H11AuO5S in the portion of Injectable Suspension taken. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS OTHER REQUIREMENTS: Injections 1. It meets the requirements under
Avobenzone
(Comment on this Monograph)id=m6560=Avobenzone=AMonos.pdf)
310.39 C20H22O3 1,3-Propanedione, 1-[4-(1,1-dimethylethyl)phenyl]-3-(4methoxyphenyl)-; 1-(p-tert-Butylphenyl)-3-(p-methoxyphenyl)-1,3-propanedione. [70356-09-1]. DEFINITION Avobenzone contains NLT 95.0% and NMT 105.0% of C20H22O3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 360 nm Solution: 5 g/mL in alcohol Absorptivities: Do not differ by more than 3.0% ASSAY PROCEDURE Standard solution: 50 mg/mL of USP Avobenzone RS in acetone Sample solution: 50 mg/mL of Avobenzone in acetone Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.32-mm 25-m fused silica capillary column coated with phase G1 Temperature: See the temperature program table below.
Time (min) Injection port Detector port Column (4/min) 0 20 Temperature 200 280 200 280
Carrier gas: Helium Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.0 between avobenzone and any adjacent peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C20H22O3 in the portion taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Avobenzone RS in the Standard solution (mg per mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass. Protect from light. USP REFERENCE STANDARDS 11 USP Aurothioglucose RS
USP 32
= concentration of avobenzone in the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% IMPURITIES Organic Impurities PROCEDURE Sample solution: Proceed as directed for the Sample solution in the Assay. Chromatographic system: Proceed as directed in the Assay. Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of C20H22O3 taken: Result = (rU/rT) 100 = peak response for each impurity, other than the avobenzone peak, of the Sample solution = sum of all of the peak responses of the Sample rT solution Acceptance criteria Individual impurities: NMT 3.0% Sum of all of the impurities: NMT 4.5% rU SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 8186 LOSS ON DRYING 731: Dry in vacuum at 70 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Avobenzone RS CU
Official Monographs / Azaperone 315

Azaperone
(Comment on this Monograph)id=m6570=Azaperone=AMonos.pdf)
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Adsorbent: 0.2-mm layer of chromatographic silica gel mixture with chemically bonded amino groups Allow the spots to dry. Standard solution A: 0.50 mg/mL of USP Azaperone RS in acetone and methylene chloride (5:1) Standard solution B: Quantitatively dilute Standard solution A with a (5:1) mixture of acetone and methylene chloride to obtain a 0.25 mg/mL solution of USP Azaperone RS. Sample solution: 50 mg/mL of Azaperone in acetone and methylene chloride (5:1) Application volume: 1 L Developing solvent system: Cyclohexane, acetone, and methanol (13:6:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Analysis: Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatograms in the Developing solvent system, until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chromatographic chamber, and allow the plate to air-dry. Examine the plate under shortwavelength UV light, and compare the intensities of any secondary spots, other than any spot at the origin, observed in the chromatogram of the Sample solution with those of the principal spots in the chromatograms of Standard solution A and Standard solution B. Acceptance criteria: The sum of the intensities of the secondary spots of the Sample solution corresponds to not more than the intensity of the principal spot of Standard solution A (1.0%). SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 9295 LOSS ON DRYING 731: Dry in vacuum at 60 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, protected from light. Store at room temperature. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Azaperone RS
327.40 C19H22FN3O 1-Butanone, 1-(4-fluorophenyl)-4-[4-(2-pyridinyl)-1-piperazinyl]-; 4-Fluoro-4-[4-(2-pyridyl)-1-piperazinyl]butyrophenone [1649-18-9]. DEFINITION Azaperone contains NLT 98.0% and NMT 102.0% of C19H22FN3O, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K: Previously dried
Azaperone Injection
(Comment on this Monograph)id=m6575=Azaperone Injection=A-Monos.pdf) DEFINITION Azaperone Injection is a sterile solution of Azaperone in Water for Injection, prepared with the aid of Tartaric Acid. It may contain a suitable preservative and a stabilizing agent. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C19H22FN3O. IDENTIFICATION The chromatogram of the Sample solution exhibits a major peak for azaperone, the retention time of which corresponds to that of the Standard solution, as obtained in the Assay.
ASSAY PROCEDURE Sample: 120 mg of Azaperone Analysis: Dissolve in 50 mL of a mixture of methyl ethyl ketone and glacial acetic acid (7:1). Add 3 drops of pnaphtholbenzein TS, and titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 16.37 mg of C19H22FN3O.
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Azaperone / Official Monographs
USP 32
ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.01 M dibasic potassium phosphate mixture (3:2) Adjust by the addition of dilute phosphoric acid (1 in 10) to a pH of 7.8 0.1. Internal standard solution: 0.5 mg/mL of benzophenone in methanol Standard stock solution: 0.5 mg/mL of USP Azaperone RS in methanol Standard solution: Combine 2.5 mL of Standard stock solution and 2.5 mL of Internal standard solution, and dilute quantitatively with methanol to 10.0 mL. Sample stock solution: 0.5 mg/mL of azaperone from Injection in methanol Sample solution: Combine 2.5 mL of Sample stock solution and 2.5 mL of Internal standard solution, and dilute quantitatively with methanol to 10.0 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 243 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.7 between the azaperone and the internal standard peaks, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H22FN3O in each mL of the Injection taken: Result = (RU/RS) (CS/CU) 100 = ratio of the azaperone peak to the benzophenone peak of the Sample solution = ratio of the azaperone peak to the RS benzophenone peak of the Standard solution = concentration of USP Azaperone RS in the CS Standard solution (mg/mL) = concentration of azaperone in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS PH 791: 4.05.6 OTHER REQUIREMENTS: Injections 1
Azatadine Maleate
(Comment on this Monograph)id=m6600=Azatadine Maleate=A-Monos.pdf)
522.55 C20H22N2 2C4H4O4 5H-Benzo[5,6]cyclohepta[1,2-b]pyridine, 6,11-dihydro-11-(1methyl-4-piperidinylidene)-, (Z)-2-butenedioate (1:2); 6,11-Dihydro-11-(1-methyl-4-piperidylidene)-5H-benzo [5,6]cyclohepta[1,2-b]pyridine maleate (1:2) [3978-86-7]. DEFINITION Azatadine Maleate contains NLT 98.0% and NMT 102.0% of C20H22N2 2C4H4O4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Solution: 40 g/mL Medium: 0.25 N hydrochloric acid in methanol ASSAY PROCEDURE Sample solution: Dissolve about 650 mg of Azatadine Maleate in 50 mL of glacial acetic acid, and add 2 drops of crystal violet TS. Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 26.13 mg of C20H22N2 2C4H4O4. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Sample solution: 7 mg/mL of the Azatadine Maleate in a mixture of toluene and methanol (1:1) Standard solution: 7 mg/mL of USP Azatadine Maleate RS in a mixture of toluene and methanol (1:1) Application volume: 100 L Developing solvent system: Toluene, isopropyl alcohol, and diethylamine (10:10:1) Analysis 1 Samples: Sample solution and Standard solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by visualization under short-wavelength UV light.
Meets the requirements under
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, protected from light. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Azaperone RS
USP 32
Separately transfer the silica gel mixture containing the principal spot from each track to suitable stoppered centrifuge tubes. Similarly transfer an equal amount of silica gel from a blank section of the plate to a separate, suitable stoppered centrifuge tube. To each of the three tubes, add 15.0 mL of a solvent mixture consisting of methanol and 0.5 N hydrochloric acid (4:1), shake vigorously for about 15 min, centrifuge, and use the supernatants for the next spectrometric Analysis. [NOTETake care to separate the principal spots from any adjacent spots.] Spectrometric conditions Cell: 1 cm Analytical wavelength: At 284 nm Blank: Solution obtained from the blank section of the plate Analysis 2 Samples: Sample solution and the Standard solution Concomitantly determine the absorbances. Calculate the chromatographic purity in the portion of Azatadine Maleate taken: (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Azatadine Maleate RS in the Standard solution (mg/mL) = nominal concentration of azatadine maleate in CU the Sample solution (mg/mL) Acceptance criteria: NLT 98.0% SPECIFIC TESTS LOSS ON DRYING 731: Dry it in vacuum at 60 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Azatadine Maleate RS AU AS CS
Official Monographs / Azatadine 317

solvent hexane extracts on a steam bath under a stream of nitrogen to dryness, pipet 1 mL of solvent hexane into each flask, insert the stoppers, and mix by use of a vortex mixer (or equivalent) until the residues have dissolved. Use these solutions as the Standard solution and the Sample solution, respectively. Apply 100 L of each of the Sample and Standard solutions separately to a suitable thin-layer chromatographic plate. Allow the spots to dry, and develop the chromatogram in the Developing solvent system, until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the plate to air-dry. Examine the plate under short-wavelength UV light. Acceptance criteria: The RF value and intensity of the principal spot in the chromatogram of the Sample solution correspond to those from the Standard solution. ASSAY PROCEDURE Standard solution: 0.06 mg/mL of USP Azatadine Maleate RS in 0.1 N hydrochloric acid Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, equivalent to 1.5 mg of azatadine maleate, to a 50-mL flask fitted with a glass stopper. Add 25.0 mL of 0.1 N hydrochloric acid, insert the stopper, and shake the mixture by mechanical means for 30 min. Filter the mixture into a suitable glass-stoppered vessel, discarding the first 5 mL of the filtrate (0.06 mg/mL of azatadine maleate). Analysis: Separately transfer 15.0 mL of the Standard solution, 15.0 mL of the Sample solution, and 15.0 mL of 0.1 N hydrochloric acid to provide the reagent blank to three 50-mL centrifuge tubes fitted with glass stoppers. To each centrifuge tube, add 10.0 mL of 1.0 N sodium hydroxide and 20 mL of solvent hexane, insert the stoppers, rotate the centrifuge tubes for 15 min, and centrifuge until the supernatants (solvent hexane phase) are clear. With the aid of separate syringes, transfer the supernatants to separate 50-mL centrifuge tubes fitted with glass stoppers. Rinse each syringe with 10 mL of solvent hexane, and add the rinse to the aqueous phase from which the respective supernatant was removed. Insert the stoppers, rotate each tube for 10 min, and centrifuge. Transfer each supernatant to the respective supernatant previously collected. Pipet 15 mL of 0.1 N hydrochloric acid into each centrifuge tube containing the combined supernatants, insert the stoppers, rotate each tube for 15 min, and centrifuge. Remove and discard the supernatants. Concomitantly determine the absorbances of the solutions. Spectrometric conditions Cell: 1 cm Analytical wavelength: At 283 nm Analysis Samples: Standard solution, Sample solution, and blank [NOTEUsing the prepared reagent blank, zero the spectrophotometer with 0.1 N hydrochloric acid.] Calculate the percentage of C20H22N2 2C4H4O4 in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Azatadine Maleate RS in the Standard solution (mg/mL) = nominal concentration of azatadine maleate in the Sample solution (mg/mL)
Azatadine Maleate Tablets

(Comment on this Monograph)id=m6630=Azatadine Maleate Tablets=A-Monos.pdf) DEFINITION Azatadine Maleate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C20H22N2 2C4H4O4. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 100 L Developing solvent system: Toluene, diethylamine, and isopropyl alcohol (10:1:10) Analysis Samples: Standard solution and Sample solution Transfer 15.0 mL of the Standard solution and 15.0 mL of the Sample solution, respectively, prepared as directed in the Assay, to separate 50-mL centrifuge tubes fitted with glass stoppers. To each centrifuge tube, add 10.0 mL of 1.0 N sodium hydroxide and 20 mL of solvent hexane, insert the stoppers, rotate the centrifuge tubes for 15 min, and centrifuge. Transfer the solvent hexane extracts (upper phase) from each centrifuge tube to separate 50mL conical flasks fitted with glass stoppers. Evaporate the
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Azatadine / Official Monographs

90.0%110.0%
USP 32
Application volume: 5 L Analysis Samples: Sample solution, Standard solution A, and Standard solution B Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Apply the Samples at points 2 cm from the bottom edge of a thin-layer chromatographic plate. Allow the spots to dry, and develop the chromatogram in a suitable chamber, using butyl alcohol, previously saturated with 6 N ammonium hydroxide, as the solvent, until the solvent front has moved 15 cm from the point of application. Remove the plate, air-dry, and locate the spots by viewing under short- and long-wavelength UV light. Acceptance criteria: Any spot from Azathioprine, other than the principal spot, is not more intense than the spot from USP Mercaptopurine RS (1.0%). SPECIFIC TESTS ACIDITY OR ALKALINITY: Shake 2.0 g with 100 mL of water for 15 min, and filter: 20.0 mL of the filtrate requires for neutralization NMT 0.10 mL of 0.020 N hydrochloric acid or NMT 0.10 mL of 0.020 N sodium hydroxide, methyl red TS being used as the indicator. LOSS ON DRYING 731: Dry it in vacuum at 105 for 5 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Azathioprine RS USP Mercaptopurine RS
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 283 nm Standard solution: USP Azatadine Maleate RS in Medium Sample solutions: Sample per Dissolution 711 Dilute with Medium to a concentration that is similar to that of the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C20H22N2 2C4H4O4 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Azatadine Maleate RS
Azathioprine
(Comment on this Monograph)id=m6690=Azathioprine=AMonos.pdf)
277.26 C9H7N7O2S 1H-Purine, 6-[(1-methyl-4-nitro-1H-imidazol-5-yl)thio]-; 6-[(1-Methyl-4-nitroimidazol-5-yl)thio]purine [446-86-6]. DEFINITION Azathioprine contains NLT 98.0% and NMT 101.5% of C9H7N7O2S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The principal spot from the Sample solution in the test for Limit of mercaptopurine shows the same RF value as that obtained from the solution of USP Azathioprine RS. ASSAY PROCEDURE Sample: 300 mg of Azathioprine Analysis: Dissolve in 80 mL of dimethylformamide. Add 5 drops of a 1 in 100 solution of thymol blue in dimethylformamide. Titrate with 0.1 N tetrabutylammonium hydroxide VS, using a magnetic stirrer, and taking precautions to prevent absorption of atmospheric carbon dioxide. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N tetrabutylammonium hydroxide is equivalent to 27.73 mg of C9H7N7O2S. Acceptance criteria: 98.0%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities LIMIT OF MERCAPTOPURINE Adsorbent: 0.25-mm layer of microcrystalline cellulose Standard solution A: 20 mg/mL of USP Azathioprine RS in 6 N ammonium hydroxide Standard solution B: 200 g/mL of USP Mercaptopurine RS, on the anhydrous basis in 6 N ammonium hydroxide Sample solution: 20 mg/mL of Azathioprine in 6 N ammonium hydroxide
Azathioprine Oral Suspension

(Comment on this Monograph)id=m1378=Azathioprine Oral Suspension=A-Monos.pdf) DEFINITION Azathioprine Oral Suspension contains NLT 90.0 %and NMT 110.0 %of the labeled amount of azathioprine (C9 H7 N7 O2 S). Prepare Azathioprine Oral Suspension 50 mg/mL as follows (See Pharmaceutical CompoundingNonsterile Solutions 795):
Azathioprine Vehicle: a mixture of Vehicle for Oral Solution, (regular or sugar-free), NF and Vehicle for Oral Suspension, NF (1:1) To make 5g A sufficient quantity
100 mL
If using Tablets, comminute them to a fine powder in a suitable mortar, or add Azathioprine powder to the mortar. Add about 10 mL of the Vehicle, and mix to a uniform paste. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add sufficient Vehicle to bring to final volume, and mix well. [CAUTIONAvoid skin contact or inhalation of azathioprine by using protective gloves and a fume hood or surgical mask.] ASSAY PROCEDURE Mobile phase: Dissolve 1.1 g of sodium-1-heptanesulfonate in 700 ml water, add 30 mL of methanol. Adjust with 1 N hydrochloric acid to a pH of 3.5. Standard solution: 25 mg of USP Azathioprine RS, in a 50mL volumetric flask
USP 32
Add 15 mL of methanol and 0.5 mL of ammonium hydroxide to the flask, swirl and sonicate for 2 mins. Dilute with methanol to volume. Transfer 10 mL of this solution to a 50-mL volumetric flask, and dilute with water to volume. Sample solution: Agitate the container of Oral Suspension for 30 mins on a rotating mixer, remove a 5-mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the sample from the freezer, allow it to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 1.0 mL of the Sample solution into a 100-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 1.3% Retention time: 4 min Analysis Samples: Sample solution and Standard solution Calculate the percentage of C9H7N7O2S in the volume of Oral Suspension : Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Azathioprine RS in the Standard solution (g/mL) = nominal concentration of the Sample solution CU (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 3.84.8 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at room temperature, or in a cold place. LABELING: Label it to state that it is to be well shaken before use, and to state the beyond-use date. Beyond-use date: 60 days after the day on which it was compounded USP REFERENCE STANDARDS 11 USP Azathioprine RS rU rS CS
Official Monographs / Azathioprine 319

Filter the solution. Chromatographic system Adsorbent: 0.25-mm layer of microcrystalline cellulose Application volume: 5 L Developing solvent system: Butyl alcohol, previously saturated with 6 N ammonium hydroxide Analysis Samples: Standard solution and Sample solution Proceed as directed in the chapter. ASSAY PROCEDURE Mobile phase: To 1.1 g of sodium 1-heptanesulfonate in 700 mL of water, add 300 mL of methanol. Adjust the solution with 1 N hydrochloric acid to a pH of 3.5. Filter the solution through a 0.8-m solvent-resistant membrane, and degas. Standard stock solution: Transfer 25 mg of USP Azathioprine RS to a 50-mL volumetric flask. Add 15 mL of methanol and 0.5 mL of ammonium hydroxide to the flask, swirl, and sonicate for 2 min. Dilute with methanol to volume. Standard solution: Transfer 10.0 mL of Standard stock solution to a 50-mL volumetric flask, and dilute with water to volume. Sample stock solution: Finely powder NLT 20 Tablets. Weigh a portion of the powder, equivalent to 50 mg of azathioprine, and transfer to a 100-mL volumetric flask. Add 25 mL of methanol and 1.0 mL of ammonium hydroxide to the flask, swirl, and sonicate for 2 min. Dilute with methanol to volume. Allow the excipients to settle. Sample solution: Transfer 10.0 mL of the Sample stock solution to a 50-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 800 theoretical plates, Standard solution Tailing factor: NMT 1.5 for the azathioprine peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H7N7O2S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response for azathioprine from the Sample solution = peak response for azathioprine from the Standard solution = concentration of USP Azathioprine RS in the Standard solution (mg/mL) = concentration of azathioprine in the Sample solution (mg/mL)
Azathioprine Tablets
(Comment on this Monograph)id=m6720=Azathioprine Tablets=A-Monos.pdf) DEFINITION Azathioprine Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of azathioprine (C9H7N7O2S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 20 mg/mL of USP Azathioprine RS in 6 N ammonium hydroxide Sample solution: Equivalent to 20 mg/mL of azathioprine, from Powdered Tablets in 6 N ammonium hydroxide
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Azathioprine / Official Monographs

93.0%107.0% CS
USP 32
= concentration of USP Azathioprine RS in the Standard solution (mg/mL) = concentration of azathioprine in the Sample CU solution (unit/mL) L = label claim (mg/unit) Acceptance criteria: 93.0%107.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 280 nm Sample solutions: Sample per Dissolution 711 Filter and dilute, if necessary, with Medium to a concentration that is similar to that of the Standard solution. Standard solution: USP Azathioprine RS in Medium Tolerances: NLT 75% (Q) of the labeled amount of C9H7N7O2S UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Protect from light. USP REFERENCE STANDARDS 11 USP Azathioprine RS
Azathioprine Sodium for Injection

(Comment on this Monograph)id=m6730=Azathioprine Sodium for Injection=A-Monos.pdf) DEFINITION Azathioprine Sodium for Injection is a sterile solid prepared by the freeze-drying of an aqueous solution of Azathioprine and Sodium Hydroxide. It contains NLT 93.0% and NMT 107.0% of the labeled amount of azathioprine (C9H7N7O2S). IDENTIFICATION The principal spot of the Sample solution obtained in the test for Limit of mercaptopurine shows the same RF value as that obtained with Standard solution A. ASSAY PROCEDURE Standard solution: Transfer 25 mg of USP Azathioprine RS to a 50-mL volumetric flask, dissolve in 2.5 mL of 0.1 N sodium hydroxide, and dilute with water to volume. Pipet 10.0 mL of this solution into a 50-mL volumetric flask, and dilute with 0.1 N sulfuric acid to volume. Sample solution: Transfer the contents of 1 vial of Azathioprine Sodium for Injection with the aid of water to a 100-mL volumetric flask, and dilute with water to volume. Pipet 10 mL of this solution into another 100-mL volumetric flask, and dilute with 0.1 N sulfuric acid to volume. Analysis: Transfer 20 mL each of the Standard solution and the Sample solution, separately, to polarographic cells, and deaerate for 10 min with nitrogen that previously has been saturated with 0.1 N sulfuric acid. Blanket the solution with saturated nitrogen, insert the dropping mercury electrode of a suitable polarograph, and record the polarogram from 0.60 to 1.00 V, using a saturated calomel electrode as the reference electrode. Determine the height of the diffusion current as the difference between the residual current and diffusion current plateau. Calculate the percentage of C9H7N7O2S in the volume of solution taken from the vial used for the Sample solution: Result = (id)U/(id)S (CS/CU) (100/L) (id)U (id)S = diffusion currents of the Sample solution = diffusion currents of the Standard solution
IMPURITIES Organic Impurities LIMIT OF MERCAPTOPURINE Standard solution A: 10 mg/mL of USP Azathioprine RS in dimethylformamide Standard solution B: 100 g/mL of USP Mercaptopurine RS in dimethylformamide Sample solution: 10 mg/mL of Azathioprine Sodium for Injection in dimethylformamide Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Adsorbent: 250-m layer of microcrystalline cellulose Application volumes Standard solution B: 15 L Sample solution and Standard solution A: 5 L Developing solvent system: Butyl alcohol, previously saturated with 5 N ammonium hydroxide Analysis Samples: Sample solution, Standard solution A, and Standard solution B Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Apply Samples, at points 2 cm from the bottom edge of a thin-layer chromatographic plate. Allow the spots to dry, and develop the chromatogram until the solvent front has moved 15 cm from the point of application. Remove the plate, air-dry, and locate the spots by viewing under short- and long-wavelength UV light. Acceptance criteria: Any spot from azathioprine, other than the principal spot, is not more intense than the spot obtained with USP Mercaptopurine RS (3.0%). SPECIFIC TESTS COMPLETENESS OF SOLUTION 641: The contents of 1 container are soluble in 10 mL of water, to give a clear, bright yellow solution, essentially free from foreign matter. PH 791 Sample solution: The contents of 1 container dissolved in 10 mL of water Acceptance criteria: 9.811.0 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.0 USP Endotoxin Unit/mg of azathioprine. WATER DETERMINATION, Method I 921: NMT 7.0%, the Sample solution being prepared as directed for a hygroscopic specimen. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Azathioprine RS USP Endotoxin RS USP Mercaptopurine RS
USP 32
Official Monographs / Azithromycin 321

Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Amperometric electrochemical detector Detector type: Dual glassy carbon electrodes Detector mode: Oxidative screen mode Detector settings: Electrode 1 set at +0.70 0.05 V, electrode 2 set at +0.82 0.05 V, background current optimized to 85 15 nanoamperes Column Guard column: 4.6-mm 5-cm; 5-m packing L29 Analytical column: 4.6-mm 15-cm; 5-m packing L29 or 3-m packing L49 without the guard column Flow rate: 1.5 mL/min Injection size: 50 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for azaerythromycin A and azithromycin with the L29 column are 0.7 and 1.0, respectively; the relative retention times for azaerythromycin A and azithromycin with the L49 column are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between azaerythromycin A and azithromycin, System suitability solution Column efficiency: NLT 1000 theoretical plates, Standard solution Tailing factor range: 0.91.5 for azithromycin, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C38H72N2O12 in each mg of Azithromycin taken: Result = (rU/rS) (CS/CU) P = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Azithromycin RS in the Standard solution = concentration of Azithromycin in the Sample CU solution P = potency of USP Azithromycin RS (g/mg of azithromycin) Acceptance criteria: Equivalent of 9451030 g IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.3%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid HEAVY METALS, Method II 231: NMT 25 ppm Organic Impurities PROCEDURE 1 [NOTEPerform either Procedure 1 or Procedure 2, depending on the manufacturing process used.] [NOTEUse water that has a resistivity of NLT 18 Mohmcm.] Mobile phase: Proceed as directed in the Assay. Solution A: 2.7 mg/mL of monobasic potassium phosphate in water Adjust with 10 N potassium hydroxide to a pH of 7.5 0.1. Diluent: Acetonitrile and Solution A (1:3) Standard stock solution: 45 g/mL of USP Desosaminylazithromycin RS, 105 g/mL of USP NDemethylazithromycin RS, and 160 g/mL of USP Azithromycin RS in acetonitrile Standard solution: 0.9 g/mL of USP Desosaminylazithromycin RS, 2.1 g/mL of USP NrU rS CS
Azithromycin
(Comment on this Monograph)id=m6740=Azithromycin=AMonos.pdf)
749.00 C38H72N2O12 xH2O (anhydrous) [83905-01-5]. 1-Oxa-6-azacyclopentadecan-15-one, 13-[(2,6-dideoxy-3-Cmethyl-3-O-methyl--L-ribo-hexopyranosyl)oxy]-2ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-heptamethyl-11-[ [3,4,6-trideoxy-3-(dimethylamino)--D-xylohexopyranosyl]oxy]- [2R(2R*,3S*,4R*,5R*,8R*,10R*,11R*,12S*, 13S*,14R*)]; (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-Cmethyl-3-O-methyl--L-ribo-hexopyranosyl)oxy]-2ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-heptamethyl-11-[ [3,4,6-trideoxy-3-(dimethylamino)--D-xylohexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one; 9-Deoxo-9a-aza-9a-methyl-9a-homoerythromycin A Monohydrate 767.02 [121479-24-4]. Dihydrate 785.02 [117772-70-0]. DEFINITION Azithromycin is anhydrous or contains one or two molecules of water of hydration. It contains the equivalent of NLT 945 g and NMT 1030 g of azithromycin (C38H72N2O12) per mg, calculated on the anhydrous basis. [NOTEUse water that has a resistivity of NLT 18 Mohm-cm (0.055S-cm) for all tests in this monograph.] IDENTIFICATION A. INFRARED ABSORPTION 197K: If a difference appears in the IR spectra of the analyte and the standard, dissolve equal portions of the test specimen and the Reference Standard in equal volumes of methanol. Evaporate the solutions to dryness on a water bath, and dry at 80 for 30 min under vacuum. Perform the test on the residues. B. The retention time of the azithromycin peak in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 5.8 g of monobasic potassium phosphate in 2130 mL of water, add 870 mL of acetonitrile. Adjust with 6 mL of 10 N potassium hydroxide to a pH of 11.0 0.1 and filter. Standard stock solution: 0.165 mg/mL of USP Azithromycin RS in acetonitrile Standard solution: 3.3 g/mL of USP Azithromycin RS, from the Standard stock solution in Mobile phase Sample stock solution: 0.165 mg/mL of Azithromycin in acetonitrile Sample solution: 3.3 g/mL of Sample stock solution in Mobile phase System suitability stock solution: 0.16 mg/mL of USP Azaerythromycin A RS in acetonitrile and Mobile phase (1:9) [NOTEDissolve in acetonitrile by swirling and with the aid of brief sonication, and then dilute with Mobile phase to volume.] System suitability solution: Transfer 2.0 mL of the System suitability stock solution and 2.0 mL of the Standard stock solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume.
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Acceptance criteria: See Impurity Table 1.
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Demethylazithromycin RS, and 3.2 g/mL of USP Azithromycin RS from Standard stock solution in Diluent Sample solution: 33 mg of Azithromycin in a 100-mL volumetric flask Add 5 mL of acetonitrile, and sonicate for about 20 s to dissolve. Dilute with Diluent to volume. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Amperometric electrochemical detector electrodes Detector type: Dual glassy carbon Detector mode: Oxidative screen mode Detector settings: Electrode 1 set at +0.70 0.05 V, electrode 2 set at +0.85 0.05 V, background current optimized to 95 25 nanoamperes [NOTEIn general, maintain electrode 1 at 0.12 V less than electrode 2, and maintain the electrodes at a constant temperature of about 26.] Column Guard column: 4.6-mm 5-cm; 5-m packing L29 Analytical column: 4.6-mm 15-cm; 5-m packing L29 or 3-m packing L49 without the guard column. Flow rate: 0.4 mL/min Injection size: 50 L System suitability Sample: Standard solution [NOTEThe relative retention times for desosaminylazithromycin, N-demethylazithromycin, and azithromycin are 0.38, 0.54, and 1.0.] Suitability requirements Column efficiency: NLT 1500 theoretical plates for the azithromycin peak Tailing factor: NMT 1.5 for each peak Relative standard deviation: NMT 5% for each of these compounds Analysis Samples: Standard solution and Sample solution [NOTERecord chromatograms for NLT 3.3 times the elution time of the azithromycin peak.] Calculate the percentages of desosaminylazithromycin and N-demethylazithromycin in the Azithromycin taken: Result = (rU/rS) (CS/CU) 100 = peak area for the relevant analyte from the Sample solution = peak area for the relevant analyte from the rS Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (g/mL) = concentration of the Sample solution CU Calculate the percentages of other related substances in the Azithromycin taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak area of each additional impurity from the Sample solution = peak area of the azithromycin peak from the Standard solution = concentration of USP Azithromycin RS in the Standard solution (g/mL) = concentration of the Sample solution (g/mL) rU
Impurity Table 1 Relative Retention Time 0.20 0.26 0.34 0.37 Acceptance Criteria (NMT %) 0.3 0.7 1.0 3.0
Name Desosaminylazithromycin N-demethylazithromycin Any other individual impurity Sum of all impurities
PROCEDURE 2 [NOTEPerform either Procedure 1 or Procedure 2, depending on the manufacturing process used.] Solution A: 8.7 mg/mL of dibasic potassium phosphate in water Adjust with 20% phosphoric acid to a pH of 8.2. Mobile phase: Acetonitrile and Solution A (3:2) Standard solution A: 35 g/mL of USP Azithromycin RS in Mobile phase Standard solution B: 7 mg/mL of USP Azithromycin Identity RS in Mobile phase Standard solution C: 14 g/mL of USP Azithromycin-NOxide RS in Mobile phase Sample solution: 7 mg/mL of Azithromycin in Mobile phase System suitability solution: 0.07 mg/mL of USP Azaerythromycin A RS and 7 mg/mL of USP Azithromycin RS in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 15-cm; 5-m packing L1 Temperature: 30 Flow rate: 0.9 mL/min Injection size: 50 L System suitability Sample: System suitability solution [NOTEFor the purpose of identification, the relative retention times for azaerythromycin A and azithromycin are 0.47 and 1.00, respectively.] Suitability requirements Resolution: NLT 8.0 between azaerythromycin A and azithromycin Tailing factor: NMT 2.5 for the azithromycin Analysis Samples: Mobile phase, Standard solution A, Standard solution B, Standard solution C, and Sample solution [NOTEDisregard any peak due to the solvent front and any peak corresponding to those obtained from the Mobile phase.] Calculate the percentage of each impurity in the portion of Azithromycin taken: Result = (rU/rS) (CS/CU) 100/RRF rU rS = peak area for each impurity from the Sample solution = peak area for each impurity from the Standard solution A
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CS CU RRF = concentration of USP Azithromycin RS in Standard solution A (mg/mL) = concentration of Sample solution (mg/mL) = relative response factor (see Impurity Table 2)
Impurity Table 2 Relative Retention Time 0.20 0.26 0.34 Relative Response Factor 0.45 1.8 4.1 Acceptance Criteria, (NMT %) 0.40 0.30 0.15

about 70, and 1.8%2.6% between the inflection point at about 70 and the inflection point at about 130. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate whether it is anhydrous, or the monohydrate or the dihydrate. The amorphous form is so labeled. Where the quantity of azithromycin is indicated in the labeling of any preparation containing Azithromycin, this shall be understood to be in terms of anhydrous azithromycin (C38H72N2O12). The labeling states with which Organic Impurities test the article complies, if other than Procedure 1. USP REFERENCE STANDARDS 11 USP Azaerythromycin A RS USP Azithromycin RS USP Azithromycin Identity RS USP Azithromycin-N-Oxide RS USP Desosaminylazithromycin RS USP N-Demethylazithromycin RS
Name Azithromycin-N-oxide 3-(N,N-didemethyl)-3-Nformylazithromycin 3-N-demethyl-3-Nformylazithromycin (rotamer 1) 3-N-demethyl-3-Nformylazithromycin (rotamer 2) 6-Demethylazithromycin (azaerythromycin A) 3-De(dimethylamino)-3oxoazithromycin 2-Desethyl-2propylazithromycin 3-Deoxyazithromycin (azithromycin B) 3-N-demethyl-3-N-[(4methylphenyl)sulfonyl]azithromycin Individual unknown impurity Total impurities
0.37
4.1
0.15
0.47 0.80 1.52 1.60 2.14
0.67 1.9 1.0 1.0 7.0
0.50 0.25 0.50 0.50 0.50
Azithromycin Capsules
(Comment on this Monograph)id=m6745=Azithromycin Capsules=A-Monos.pdf) DEFINITION Azithromycin Capsules contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of azithromycin (C38H72N2O12). IDENTIFICATION The retention time for the azithromycin peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEUse water that has a resistivity of NLT 18 Mohmcm.] Mobile phase: Dissolve 5.8 g of monobasic potassium phosphate in 2130 mL of water, and add 870 mL of acetonitrile. Adjust with 6 mL of 10 N potassium hydroxide to a pH of 11.0 0.1, and filter. Standard stock solution: 0.165 mg/mL of USP Azithromycin RS in acetonitrile [NOTEFirst dissolve in a small quantity of acetonitrile by swirling and with the aid of brief sonication, and then dilute it to volume.] Standard solution: 0.0033 mg/mL of USP Azithromycin RS, from Standard stock solution in Mobile phase Sample stock solution: Remove, as completely as possible, the contents of NLT 20 Capsules. Mix the combined contents, and transfer a quantity of the powder, equivalent to 250 mg of anhydrous azithromycin, to a 250-mL volumetric flask. Add 175 mL of acetonitrile, and shake by mechanical means for 30 min. Dilute with acetonitrile to volume. Place 40 mL of the resulting suspension in a centrifuge tube, and centrifuge. Transfer 2.0 mL of the clear supernatant to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution: Transfer 2.0 mL of Sample stock solution to a 25-mL volumetric flask, and dilute with Mobile phase to volume. System suitability stock solution: 0.16 mg/mL of USP Azaerythromycin A RS in acetonitrile and Mobile phase (1:9) [NOTEDissolve in acetonitrile by swirling and with the aid of brief sonication, and then dilute with Mobile phase to volume.] System suitability solution: Transfer 2.0 mL of the System suitability stock solution and 2.0 mL of Standard stock solution
1.0
0.20 2.0
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 45 to 49, at 20 Sample solution: 20 mg/mL, in dehydrated alcohol CRYSTALLINITY 695: Meets the requirements except, where it is labeled as amorphous, most of the particles do not exhibit birefringence and extinction positions PH 791: 9.011.0, in a mixture of methanol and water (1:1) containing 2 mg/mL, prepared by diluting a solution in methanol containing 4 mg/mL with an equal volume of water WATER DETERMINATION, Method I 921 Where it is labeled as anhydrous: NMT 2.0% Where it is labeled as the dehydrate: 4.0%5.0% Where it is labeled as the monohydrate: 1.8%4.0%, except that it may be 4.0%6.5% when the requirements of the Loss on Drying test are met LOSS ON DRYING 731 (where it is labeled as Azithromycin monohydrate and has a water content of 4.0%6.5%) (See Thermal Analysis 891.) [NOTEThe quantity taken for the determination may be adjusted, if necessary, for instrument sensitivity.] Procedure: Determine the percentage of volatile substances by thermogravimetric analysis in an appropriately calibrated instrument, using about 10 mg of Azithromycin. Heat the specimen at the rate of 10/min between ambient temperature and 150 in an atmosphere of nitrogen at a constant flow rate of about 35 mL/min. From the thermogram plot the derivatives of the loss on drying (percent loss per minute), identify the inflection points of the two weight loss steps at about 70 and 130. Acceptance criteria: It loses NMT 4.5% of its weight between ambient temperature and the inflection point at
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Azithromycin / Official Monographs
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solution to a second 25-mL volumetric flask, and dilute with Mobile phase to volume. Analysis: Determine the amount of C38H72N2O12 dissolved, using filtered portions of the Sample solution and using the Analysis in the Assay, making any necessary modifications. Calculate the quantity, in mg, of C38H72N2O12 dissolved: Result = 70.31 (rU/rS) (CS P) = Azithromycin peak response from the Sample solution = Azithromycin peak response from the Standard rS solution = concentration of USP Azithromycin RS in the CS Standard solution (mg/mL) P = potency of USP Azithromycin RS (g/mg) Tolerances: NLT 75% (Q) of the labeled amount of azithromycin is dissolved in 45 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 5.0%
to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Amperometric electrochemical detector with dual glassy carbon electrodes operated in the oxidative screen mode with electrode 1 set at +0.70 0.05 V and electrode 2 set at +0.82 0.05 V, and the background current optimized to 85 15 nanoamperes Columns Guard column: 4.6-mm 5-cm; 5-m packing L29 Analytical column: 4.6-mm 15-cm; 5-m packing L29 or 3-m packing L49 without the guard column Flow rate: 1.5 mL/min Injection size: 50 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for azaerythromycin A and azithromycin with the L29 column are 0.7 and 1.0, respectively; the relative retention times for azaerythromycin A and azithromycin with the L49 column are 0.8 and 1.0, respectively.)] Suitability requirements Resolution: NLT 2.5 between azaerythromycin A and azithromycin, System suitability solution Column efficiency: NLT 1000 theoretical plates, Standard solution Tailing factor: NLT 0.9 for the azithromycin peak and NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C38H72N2O12 in the portion of Capsules taken: Result = (rU/rS) (CS P/CU) 100 = Azithromycin area from the Sample solution = Azithromycin area response from the Standard solution CS = concentration of USP Azithromycin RS in Standard solution (mg/mL) P = potency of USP Azithromycin RS (g/mg) = concentration of Azithromycin in Sample solution CU (g/mL) Acceptance criteria: Equivalent of 90.0%110.0% rU rS PERFORMANCE TESTS DISSOLUTION 711 [NOTEUse water that has a resistivity of NLT 18 Mohmcm.] Solution A: Prepare 6 L of 0.1 M dibasic sodium phosphate, adjust with 40 mL of hydrochloric acid to a pH of 6.0 0.05, and add 600 mg of trypsin. Medium: Solution A: 900 mL Apparatus 2: 100 rpm Time: 45 min Standard solution: Transfer 15 mg of USP Azithromycin RS to a 50-mL volumetric flask. Add 25 mL of Medium, and sonicate briefly to dissolve. Dilute with Medium to volume. Transfer 2.0 mL of this solution to a 25-mL volumetric flask, and dilute with Mobile phase (prepared as directed in the Assay) to volume. Transfer 4.0 mL of this solution to a second 25-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution: Filter a portion of the Sample solution through a filter having a porosity of 0.5-m or less. Transfer 2.0 mL of the filtrate to a 25-mL volumetric flask, and dilute with Mobile phase to volume. Transfer 4.0 mL of this
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Where packaged in unit-of-use containers, each container contains six 250-mg Capsules, and the label indicates the intended sequential day of use for each Capsule. USP REFERENCE STANDARDS 11 USP Azaerythromycin A RS USP Azithromycin RS
Azithromycin for Oral Suspension

(Comment on this Monograph)id=m6750=Azithromycin for Oral Suspension=A-Monos.pdf) DEFINITION Azithromycin for Oral Suspension is a dry mixture of Azithromycin and one or more buffers, sweeteners, diluents, anticaking agents, and flavors. It contains NLT 90.0% and NMT 110.0% of the labeled amount of azithromycin (C38H72N2O12). IDENTIFICATION The Sample solution, obtained as directed in the Assay, exhibits a major peak for azithromycin, the retention time of which corresponds to that exhibited in the chromatogram of the Standard solution, obtained as directed in the Assay. ASSAY PROCEDURE [NOTEUse water that has a resistivity of NLT 18 Mohmcm.] Mobile phase: Dissolve 5.8 g of monobasic potassium phosphate in 2130 mL of water, add 870 mL of acetonitrile. Adjust with 6 mL of 10 N potassium hydroxide to a pH of 11.0 0.1, and filter. Standard stock solution: 0.165 mg/mL of USP Azithromycin RS in acetonitrile [NOTEFirst dissolve in a small quantity of acetonitrile by swirling and with the aid of brief sonication, and then dilute it to volume.] Standard solution: 0.0033 mg/mL of USP Azithromycin RS, from Standard stock solution in Mobile phase Solvent: Dissolve 2.2 g of monobasic potassium phosphate in 1590 mL of water, add 600 mL of isopropyl alcohol, 480 mL of alcohol, and 330 mL of acetonitrile. Adjust with 10 N potassium hydroxide to a pH of 8.4 0.1, and shake by mechanical means for 30 min.
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Sample solution A (where packaged in a single-unit container): Transfer the contents of a container of Azithromycin for Oral Suspension to a volumetric flask of such capacity, V, in mL, that when diluted to volume as directed in the following two sentences the solution will contain 2 mg of azithromycin/mL. Add a volume of Solvent equal to 70% of the volume of the flask, and shake by mechanical means for 30 min. Dilute with Solvent to volume. Transfer 40 mL of this suspension to a stoppered centrifuge tube, and centrifuge for 20 min. Transfer 2.0 mL of the clear supernatant to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Transfer 2.0 mL of this solution to a second 50-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution B (where packaged in a multiple-unit container): Constitute Azithromycin for Oral Suspension as directed in the labeling. Transfer 5.0 mL of the suspension so obtained, freshly mixed and free from air bubbles, to a volumetric flask of such capacity, Vm, in mL, that when diluted to volume as directed below, the solution will contain 0.4 mg of azithromycin/mL. Add a volume of Solvent equal to 70% of the volume of the flask, and shake by mechanical means for 30 min. Dilute with Solvent to volume. Transfer 40 mL of the suspension so obtained to a stoppered centrifuge tube, and centrifuge for 20 min. Transfer 5.0 mL of the clear supernatant to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Transfer 5.0 mL of this solution to a second 50-mL volumetric flask, and dilute with Mobile phase to volume. System suitability stock solution: 0.16 mg/mL of USP Azaerythromycin A RS in acetonitrile and Mobile phase (1:9) [NOTEDissolve in acetonitrile by swirling and with the aid of brief sonication, and then dilute with Mobile phase to volume.] System suitability solution: Transfer 2.0 mL of System suitability stock solution and 2.0 mL of Standard stock solutionto a 100 mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Amperometric electrochemical detector with dual glassy carbon electrodes operated in the oxidative screen mode with electrode 1 set at +0.70 0.05 V and electrode 2 set at +0.82 0.05 V, and the background current optimized to 85 15 nanoamperes Column Guard column: 4.6-mm 5-cm; 5-m packing L29 Analytical column: 4.6-mm 15-cm; 5-m packing L29 or 3-m packing L49 without the guard column Flow rate: 1.5 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for azaerythromycin A and azithromycin with the L29 column are 0.7 and 1.0, respectively; the relative retention times for azaerythromycin A and azithromycin with the L49 column are 0.8 and 1.0, respectively.]

Suitability requirements Resolution: NLT 2.5 between azaerythromycin A and azithromycin, System suitability solution Column efficiency: NLT 1000 theoretical plates, Standard solution Tailing factor: NLT 0.9 for the azithromycin peak and NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in mg, of C38H72N2O12 in the singleunit container of Azithromycin for Oral Suspension taken: Result = (rU/rS) (125V/200) CP = Azithromycin peak response from the Sample solution A = Azithromycin peak response from the Standard rS solution V = volume, 2 mg of azithromycin/mL C = concentration of USP Azithromycin RS in the Standard solution (mg/mL) P = potency of USP Azithromycin RS (g/mg) Calculate the percentage of C38H72N2O12 in each 5 mL of suspension taken from a container of Azithromycin for Oral Suspension where packaged in a multiple-unit container: rU Result = (rU/rS) (Vm/10) CP = Azithromycin peak response from Sample solution B rS = Azithromycin peak response from the Standard solution = volume, 0.4 mg of azithromycin/mL Vm C = concentration of USP Azithromycin RS in the Standard solution (mg/mL) P = potency of USP Azithromycin RS (g/mg) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791 For solid packaged in single-unit containers: 9.011.0, in the suspension constituted as directed in the labeling For solid packaged in multiple-unit containers: 8.511.0, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 1.5% PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905: It meets the requirements for solid packaged in single-unit containers. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Azithromycin RS rU
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Aztreonam / Official Monographs
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Tailing factor: NMT 2.0 for aztreonam Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C13H17N5O8S2 in the portion of Aztreonam taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Aztreonam RS in the Standard solution (mg/mL) CU = concentration of aztreonam in the Sample solution (mg/mL) Acceptance criteria: 90.0%105.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid HEAVY METALS, Method II 231: NMT 30 ppm SPECIFIC TESTS STERILITY TESTS 71: Where the label states that Aztreonam is sterile, it meets the requirements for Test for Sterility of the Product to Be ExaminedMembrane Filtration, using Fluid A, to which 23.4 g of sterile arginine/1000 mL has been added. WATER DETERMINATION, Method I 921: NMT 2.0%; if labeled as the hydrated form: 12.0%18.0% [NOTEThe term hydrated form refers to the -form of Aztreonam, which is not a stoichiometric hydrate.] BACTERIAL ENDOTOXINS TEST 85: Where the label states that aztreonam is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.17 USP Endotoxin Unit/mg of aztreonam. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. Where it is the hydrated form, the label so indicates. USP REFERENCE STANDARDS 11 USP Aztreonam RS USP Aztreonam E-Isomer RS USP Endotoxin RS rU rS CS
Aztreonam
(Comment on this Monograph)id=m6772=Aztreonam=AMonos.pdf)
C13H17N5 O8S2 435.43 Propanoic acid, 2-[[[1-(2-amino-4-thiazolyl)-2-[(2-methyl-4oxo-1-sulfo-3-azetidinyl)amino]-2oxoethylidene]amino]oxy]-2-methyl-, [2S-[2,3(Z)]]-; (Z)-2-[[[(2-Amino-4-thiazolyl)[[(2S,3S)-2-methyl-4-oxo-1-sulfo-3azetidinyl]carbamoyl]methylene]amino]oxy]-2methylpropionic acid [78110-38-0]. DEFINITION Aztreonam is anhydrous or hydrated. The anhydrous form contains NLT 90.0% and NMT 105.0% of C13H17N5 O8S2, calculated on the as-is basis. The hydrated form contains NLT 92.0% and NMT 105.0% of C13H17N5O8S2, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K: If a difference appears in the IR spectra of the analyte and the standard, dissolve equal portions of the test specimen and the Reference Standard in equal volumes of methanol. [NOTETo achieve a complete dissolution, it is suggested to use about 25 mL of methanol for each 50 mg of material, and stir the mixture for 40 min at room temperature.] Evaporate the solutions to dryness under vacuum, and dry at 40 for 4 h under vacuum. Perform the test on the residues. ASSAY PROCEDURE Buffer: 6.8 mg/mL of monobasic potassium phosphate in water; adjust with 1 M phosphoric acid to a pH of 3.0 0.1. Mobile phase: Methanol and Buffer (1:4) Standard solution: 1 mg/mL of USP Aztreonam RS in Mobile phase System suitability solution: 0.2 mg/mL of USP Aztreonam RS and 0.2 mg/mL of USP Aztreonam E-Isomer RS in Mobile phase Sample solution: 1 mg/mL of Aztreonam in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column Guard: 2-mm 10-cm; packing L2 Analytical: 4.6-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for aztreonam and aztreonam E-isomer are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between aztreonam and aztreonam E-isomer Column efficiency: NLT 1000 theoretical plates for aztreonam
Aztreonam Injection
(Comment on this Monograph)id=m6773=Aztreonam Injection=A-Monos.pdf) DEFINITION Aztreonam Injection is a sterile solution of Aztreonam and Arginine and a suitable osmolality adjusting substance in Water for Injection. It contains NLT 90.0% and NMT 120.0% of the labeled amount of aztreonam (C13H17N5O8S2). IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay.
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ASSAY PROCEDURE Mobile phase: 1.15 g of monobasic ammonium phosphate in 800 mL of water Adjust with phosphoric acid to a pH of 2.0 0.1, and dilute with water to 1000 mL. Prepare a suitable mixture of acetonitrile and this solution (3:1). Standard solution: 0.2 mg/mL each of USP Aztreonam RS and USP L-Arginine RS, in Mobile phase System suitability solution: 0.2 mg/mL of each, USP Aztreonam RS and USP Open Ring Aztreonam RS, in Mobile phase Sample solution: 0.2 mg/mL of aztreonam, from volume of Injection in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 206 nm Column: 4.6-mm 50-cm saturator precolumn containing packing L27, and a 4-mm 25-cm analytical column containing packing L20 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for aztreonam and open ring aztreonam are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between aztreonam and open ring aztreonam in the System suitability solution Column efficiency: NLT 1000 theoretical plates determined from the aztreonam peak, System suitability solution Tailing factor: NMT 2.0 for the aztreonam peak, System suitability solution Relative standard deviation: NMT 2.0%, System suitability solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C13H17N5O8S2 in the volume of Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Aztreonam RS in the Standard solution (mg/mL) = concentration of aztreonam in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%120.0% rU rS CS SPECIFIC TESTS PYROGEN TEST 151: It meets the requirements, the test dose being an accurately measured volume of Injection sufficient to provide 50 mg of aztreonam per kg. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 4.57.5 PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. Maintain in the frozen state. LABELING: It meets the requirements for Injections 1, Labels and Labeling. The label states that it is to be thawed just prior to use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen.
Official Monographs / Aztreonam 327

USP REFERENCE STANDARDS 11 USP L-Arginine RS USP Aztreonam RS USP Open Ring Aztreonam RS
Aztreonam for Injection

(Comment on this Monograph)id=m6781=Aztreonam for Injection=A-Monos.pdf) DEFINITION Aztreonam for Injection is a dry mixture of sterile Aztreonam and Arginine. It contains NLT 90.0% and NMT 105.0% of aztreonam (C13H17N5O8S2), calculated on the anhydrous and arginine-free basis. Each container contains NLT 90.0% and NMT 120.0% of the labeled amount of aztreonam (C13H17N5O8S2). IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 1.15 g of monobasic ammonium phosphate in 800 mL of water Adjust with phosphoric acid to a pH of 2.0 0.1, and dilute with water to 1000 mL. Prepare a suitable mixture of acetonitrile and this solution (3:1). Standard solution: 0.2 mg/mL each of USP Aztreonam RS and USP L-Arginine RS, in Mobile phase System suitability solution: 0.2 mg/mL each of USP Aztreonam RS and USP Open Ring Aztreonam RS in Mobile phase Sample solution A: Weigh one container of Aztreonam for Injection. Transfer the contents of the container to a 100-mL volumetric flask. Weigh the empty container, and calculate the weight, in mg, of Aztreonam for Injection removed. Dissolve the powder in the volumetric flask in Mobile phase, and dilute with Mobile phase to volume. Dilute a volume of this solution with Mobile phase to obtain a solution having a concentration of 0.2 mg/mL of aztreonam. Sample solution B: Constitute one container of Aztreonam for Injection with a volume of water, corresponding to the volume of solvent specified in the labeling, except where the capacity of the container is 100 mL or greater to constitute with 10 mL of water. Withdraw the total withdrawable contents of the container, and dilute with Mobile phase to obtain a solution containing 0.2 mg/mL of aztreonam. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 206 nm Column: 4.6-mm 50-cm saturator precolumn containing packing L27, and a 4-mm 25-cm analytical column containing packing L20 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for aztreonam and open ring aztreonam are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between aztreonam and open ring aztreonam, System suitability solution Column efficiency: NLT 1000 theoretical plates determined from the aztreonam peak, System suitability solution
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Aztreonam / Official Monographs
USP 32
Sample solution: Use Sample solution A, prepared as directed in the Assay. Analysis: Proceed as directed in the Assay. Calculate the percentage of C6H14N4O2 in the Aztreonam for Injection taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of USP L-Arginine RS in the Standard solution (mg/mL) = concentration of Aztreonam for Injection in CU Sample solution A, based on the weight of Aztreonam for Injection removed from the container (mg) and the extent of dilution (mg/mL) Use this percentage to calculate, on the anhydrous and arginine-free basis, the Assay result from Sample solution A, obtained as directed in the Assay.
Tailing factor: NMT 2.0 for the aztreonam peak, System suitability solution Relative standard deviation: NMT 2.0%, System suitability solution Analysis Samples: Standard solution, Sample solution A, and Sample solution B [NOTEThe relative retention times for aztreonam and arginine are 0.3 and 1.0, respectively.] Calculate the percentage of C13H17N5O8S2 in the Aztreonam for Injection taken: Result = 0.1(rU/rS) (CS PS/CU) = peak response from Sample solution A = peak response from the Standard solution = concentration of USP Aztreonam RS in the Standard solution (mg/mL) = assigned purity of USP Aztreonam RS (g/mg) PS = concentration of Aztreonam for Injection in CU Sample solution A (mg/mL), based on the weight of Aztreonam for Injection removed from the container (mg) and the extent of dilution Calculate the quantity, in mg, of C13H17N5O8S2 in the container of Aztreonam for Injection used to prepare Sample solution B: Result = (rU/rS) (CS PS L/1000 CU) = peak response from Sample solution B = peak response from the Standard solution = concentration of USP Aztreonam RS in the Standard solution (mg/mL) = assigned purity of USP Aztreonam RS (g/mg) PS L = labeled quantity of aztreonam in the container of Aztreonam for Injection (mg) = concentration of aztreonam in Sample solution B CU (mg/mL), on the basis of the labeled quantity of aztreonam in the container (mg) and the extent of dilution Acceptance criteria: 90.0%105.0% Each container contains 90.0%120.0% of the labeled amount of C13H17N5O8S2. rU rS CS OTHER COMPONENTS CONTENT OF ARGININE Mobile phase, Standard solution, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. rU rS CS
SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.17 USP Endotoxin Unit/mg of aztreonam. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791 Sample solution: 100 mg/mL Acceptance criteria: 4.57.5 WATER DETERMINATION, Method I 921: NMT 2.0% PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: It meets the requirements for Injections 1, Labeling. PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP L-Arginine RS USP Aztreonam RS USP Endotoxin RS USP Open Ring Aztreonam RS
USP 32
Bacampicillin Hydrochloride Bacampicillin Hydrochloride for Oral Suspension Bacampicillin Hydrochloride Tablets Bacitracin Bacitracin for Injection Bacitracin Ointment Bacitracin Ophthalmic Ointment Bacitracin and Polymyxin B Sulfate Topical Aerosol Soluble Bacitracin Methylene Disalicylate Bacitracin Methylene Disalicylate Soluble Powder Bacitracin Zinc Bacitracin Zinc Ointment Bacitracin Zinc Soluble Powder Bacitracin Zinc and Polymyxin B Sulfate Ointment Bacitracin Zinc and Polymyxin B Sulfate Ophthalmic Ointment Baclofen Baclofen Oral Suspension Baclofen Tablets Adhesive Bandage Gauze Bandage Barium Hydroxide Lime Barium Sulfate Barium Sulfate Paste Barium Sulfate Suspension Barium Sulfate for Suspension Barium Sulfate Tablets BCG Live BCG Vaccine Beclomethasone Dipropionate Belladonna Leaf Belladonna Extract Belladonna Extract Tablets Belladonna Tincture Benazepril Hydrochloride Benazepril Hydrochloride Tablets Bendroflumethiazide Bendroflumethiazide Tablets Benoxinate Hydrochloride Benoxinate Hydrochloride Ophthalmic Solution Benzethonium Chloride Benzethonium Chloride Concentrate Benzethonium Chloride Topical Solution
MONOGRAPH LIST /
Benzethonium Chloride Tincture Benzocaine Benzocaine Topical Aerosol Benzocaine Cream Benzocaine Gel Benzocaine Lozenges Benzocaine Ointment Benzocaine Otic Solution Benzocaine Topical Solution Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Aerosol Benzocaine, Butamben, and Tetracaine Hydrochloride Gel Benzocaine, Butamben, and Tetracaine Hydrochloride Ointment Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Solution Benzocaine and Menthol Topical Aerosol Benzoic Acid Benzoic and Salicylic Acids Ointment Benzoin Compound Benzoin Tincture Benzonatate Benzonatate Capsules Hydrous Benzoyl Peroxide Benzoyl Peroxide Gel Benzoyl Peroxide Lotion Benztropine Mesylate Benztropine Mesylate Injection Benztropine Mesylate Tablets Benzyl Benzoate Benzyl Benzoate Lotion Benzylpenicilloyl Polylysine Concentrate Benzylpenicilloyl Polylysine Injection Beta Carotene Beta Carotene Capsules Betahistine Hydrochloride Betaine Hydrochloride Betamethasone Betamethasone Cream Betamethasone Oral Solution Betamethasone Tablets Betamethasone Acetate Betamethasone Benzoate Betamethasone Benzoate Gel
/ MONOGRAPH LIST
Bismuth Subnitrate Bismuth Subsalicylate Bismuth Subsalicylate Magma Bismuth Subsalicylate Oral Suspension Bismuth Subsalicylate Tablets Bisoctrizole Bisoprolol Fumarate Bisoprolol Fumarate Tablets
USP 32
Betamethasone Dipropionate Betamethasone Dipropionate Topical Aerosol Betamethasone Dipropionate Cream Betamethasone Dipropionate Lotion Betamethasone Dipropionate Ointment Betamethasone Sodium Phosphate Betamethasone Sodium Phosphate Injection Betamethasone Sodium Phosphate and Betamethasone Acetate Injectable Suspension Betamethasone Valerate Betamethasone Valerate Cream Betamethasone Valerate Lotion Betamethasone Valerate Ointment Betaxolol Hydrochloride Betaxolol Ophthalmic Solution Betaxolol Tablets Bethanechol Chloride Bethanechol Chloride Injection Bethanechol Chloride Oral Solution Bethanechol Chloride Oral Suspension Bethanechol Chloride Tablets Bicalutamide Bicalutamide Tablets Biological Indicator for Dry-Heat Sterilization, Paper Carrier Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Liquid Spore Suspensions Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Nonpaper Carriers Biological Indicator for Steam Sterilization, Paper Carrier Biological Indicator for Steam Sterilization, Self-Contained Biotin Biperiden Biperiden Hydrochloride Biperiden Hydrochloride Tablets Biperiden Lactate Injection Bisacodyl Bisacodyl Suppositories Bisacodyl Rectal Suspension Bisacodyl Delayed-Release Tablets Milk of Bismuth Bismuth Citrate Bismuth Subcarbonate Bismuth Subgallate
Bisoprolol Fumarate and Hydrochlorothiazide Tablets Bleomycin Sulfate Bleomycin for Injection Anti-A Blood Grouping Serum Anti-B Blood Grouping Serum Blood Grouping Serums Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e Whole Blood Red Blood Cells Platelets Botulism Antitoxin Bovine Acellular Dermal Matrix Bretylium Tosylate Bretylium Tosylate Injection Bretylium Tosylate in Dextrose Injection Brinzolamide Brinzolamide Ophthalmic Suspension Bromocriptine Mesylate Bromocriptine Mesylate Capsules Bromocriptine Mesylate Tablets Bromodiphenhydramine Hydrochloride Bromodiphenhydramine Hydrochloride Oral Solution Bromodiphenhydramine Hydrochloride and Codeine Phosphate Oral Solution Brompheniramine Maleate Brompheniramine Maleate Injection Brompheniramine Maleate Oral Solution Brompheniramine Maleate Tablets Brompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution Budesonide Bumetanide Bumetanide Injection Bumetanide Tablets Bupivacaine Hydrochloride Bupivacaine Hydrochloride Injection
USP 32
Bupivacaine Hydrochloride in Dextrose Injection Bupivacaine Hydrochloride and Epinephrine Injection Buprenorphine Hydrochloride Bupropion Hydrochloride Bupropion Hydrochloride Tablets Bupropion Hydrochloride Extended-Release Tablets Buspirone Hydrochloride Buspirone Hydrochloride Tablets Busulfan Busulfan Tablets Butabarbital Butabarbital Sodium Butabarbital Sodium Oral Solution Butabarbital Sodium Tablets Butalbital
MONOGRAPH LIST /
Butalbital, Acetaminophen, and Caffeine Capsules Butalbital, Acetaminophen, and Caffeine Tablets Butalbital and Aspirin Tablets Butalbital, Aspirin, and Caffeine Capsules Butalbital, Aspirin, and Caffeine Tablets Butalbital, Aspirin, Caffeine, and Codeine Phosphate Capsules Butamben Butoconazole Nitrate Butoconazole Nitrate Vaginal Cream Butorphanol Tartrate Butorphanol Tartrate Injection Butorphanol Tartrate Nasal Solution
USP 32
Official Monographs / Bacampicillin 1

Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3000 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g/mg, of the ampicillin (C16H19N3O4S) equivalent in the portion of Bacampicillin Hydrochloride taken: Result = (rU/rS) (CS/CU) P (Mr1/Mr2) = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Bacampicillin RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU P = potency of USP Bacampicillin Hydrochloride RS (g of ampicillin/mg of RS) = molecular weight of anhydrous ampicillin, Mr1 349.41 = molecular weight of bacampicillin hydrochloride, Mr2 501.98 Acceptance criteria: 623727 g ampicillin in each mg of Bacampicillin Hydrochloride rU rS CS IMPURITIES Organic impurities DIMETHYLANILINE 223: Meets the requirements Sample solution: Transfer 1.0 g of the sample to a suitable centrifuge tube, add 5.0 mL of 2 N sodium hydroxide, swirl to dissolve the specimen, add 1.0 mL of Internal Standard Solution prepared as directed in the chapter, shake vigorously for 1 min, and centrifuge. Use the clear supernatant. SPECIFIC TESTS PH 791: 3.04.5, in a solution 20 mg/mL WATER DETERMINATION, Method I 921: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Bacampicillin Hydrochloride RS
Bacampicillin Hydrochloride
132270
(Comment on this Monograph)id=m6805=Bacampicillin Hydrochloride=B-Monos.pdf)
501.98 C21H27N3O7S HCl 4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6[(aminophenylacetyl)amino]-3,3-dimethyl-7-oxo,1[(ethoxycarbonyl)oxyethyl ester, monohydrochloride, [2S[2,5,6(S*)]]-; (2S,5R,6R)-6-[(R)-(2-Amino-2-phenylacetamido)]-3,3-dimethyl-7oxo-4-thia-1-azabicyclo[3.2.0] heptane-2-carboxylic acid ester with ethyl 1-hydroxyethyl carbonate, monohydrochloride [37661-08-8]. DEFINITION Bacampicillin Hydrochloride has a potency of NLT 623 g and NMT 727 g of ampicillin (C16H19N3O4S) per mg. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 2 mg/mL of USP Bacampicillin Hydrochloride RS in alcohol Sample solution: 2 mg/mL in alcohol Mode: TLC Application volume: 5 L Developing solvent system: Methylene chloride, chloroform, and alcohol (10:1:1) Analysis: Apply two 5-L portions of the Sample solution 4.0 cm apart. After the spots dry, apply two 5-L portions of the Standard solution, one midway between the Sample solution spots and the other to one of the Sample solution spots. When the solvent front has moved three-fourths of the length of the plate, remove the plate from the chamber, and allow to dry. Spray the plate with a spray reagent containing 1 g of ninhydrin and 1 mL of pyridine in each 100 mL of solution in butyl alcohol, and heat at 100 for 10 min. Acceptance criteria: Bacampicillin appears as a purple spot, and the RF values of the spots from the Sample solution and from the combined Sample solution and Standard solution, respectively, correspond to the RF value of the spot obtained from the Standard solution. ASSAY PROCEDURE Solution A: 0.02 M dibasic sodium phosphate. Adjust with 0.02 M monobasic sodium phosphate to a pH of 6.8 0.05. Mobile phase: Acetonitrile and Solution A (1:1), filtered Standard solution: 0.8 mg/mL of USP Bacampicillin Hydrochloride RS Sample solution: 0.8 mg/mL of Bacampicillin Hydrochloride [NOTESonicate the Standard solution and the Sample solution for about 20 min to achieve complete dissolution, and pass through a filter of 0.5 m or finer porosity.] Chromatographic system (See Chromatography 621, System Suitability.)
Bacampicillin Hydrochloride for Oral Suspension

(Comment on this Monograph)id=m6810=Bacampicillin Hydrochloride for Oral Suspension=B-Monos.pdf) DEFINITION Bacampicillin Hydrochloride for Oral Suspension contains an amount of Bacampicillin Hydrochloride equivalent to NLT 90.0% and NMT 125.0% of the labeled amount of ampicillin (C16H19N3O4S) when constituted as directed. It contains one or more suitable buffers, colors, flavors, suspending agents, and sweetening ingredients.
Bacampicillin / Official Monographs
USP 32
V = volume of constituted Sample solution taken (mL) Acceptance criteria: 90.0%125.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 Meets the requirements for Solids packaged in single-unit containers DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 6.58.0, in the suspension constituted as directed in the labeling LOSS ON DRYING 731: Dry about 100 mg in a capillarystoppered bottle in vacuum at 60 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Bacampicillin Hydrochloride RS
IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 2 mg/mL of USP Bacampicillin Hydrochloride RS in alcohol Sample solution: Equivalent to 1.4 mg/mL of ampicillin from Bacampicillin Hydrochloride for Oral Suspension in alcohol Constitute Bacampicillin Hydrochloride for Oral Suspension as directed in the labeling. Transfer a portion of the resulting suspension, equivalent to 140 mg of ampicillin, to a 100-mL volumetric flask, add 70 mL of alcohol, shake by mechanical means for 30 min, and dilute with alcohol to volume. Application volume: 5 L Developing solvent system: Methylene chloride, chloroform, and alcohol (10:1:1) Analysis Samples: Sample solution and Standard solution Apply two 5-L portions of the Sample solution 4.0 cm apart. After the spots dry, apply two 5-L portions of the Standard solution, one midway between the Sample solution spots and the other to one of the Sample solution spots, allow spots to dry and place the plate in the chamber. When the solvent front has moved three-fourths of the length of the plate, remove the plate from the chamber, and allow to dry. Spray the plate with a spray reagent containing 1 g of ninhydrin and 1 mL of pyridine in each 100 mL of solution in butyl alcohol, and heat at 100 for 10 min. Acceptance criteria: Bacampicillin appears as a purple spot, and the RF values of the spots from the Sample solution and from the combined Sample solution and Standard solution, respectively, correspond to the RF value of the spot obtained from the Standard solution. ASSAY PROCEDURE Standard solution: Using USP Ampicillin RS, prepare as directed for the Standard preparation under Iodometric AssayAntibiotics 425. Sample solution: Equivalent to 0.35 mg/mL of ampicillin from Bacampicillin Hydrochloride for Oral Suspension in alcohol and 0.1 M phosphoric acid (4:1) [NOTETransfer a volume of Bacampicillin Hydrochloride for Oral Suspension, constituted as directed in the labeling and free from bubbles, equivalent to about 87.5 mg of ampicillin, to a 250-mL volumetric flask. Add 200 mL of a solvent mixture consisting of alcohol and 0.1 M phosphoric acid (4:1). Shake by mechanical means for 30 min. Centrifuge a portion of the resulting suspension. Pipet 4.0 mL of the clear solution so obtained into each of two glass-stoppered, 125-mL conical flasks.] Analysis: Proceed as directed under Iodometric Assay Antibiotics 425, Procedure. Calculate the quantity, in mg, of C16H19N3O4S in each mL of the constituted Bacampicillin Hydrochloride for Oral Suspension taken: Result = (0.0625 F)(B I)/V F B I = microgram (or unit) equivalent = volume of 0.01 N sodium thiosulfate consumed in the Blank determination (mL) = volume of 0.01 N sodium thiosulfate consumed in the Inactivation and Titration (mL)
Bacampicillin Hydrochloride Tablets

(Comment on this Monograph)id=m6815=Bacampicillin Hydrochloride Tablets=B-Monos.pdf) DEFINITION Bacampicillin Hydrochloride Tablets contain the equivalent of NLT 90.0% and NMT 125.0% of the labeled amount of ampicillin (C16H19N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 2 mg/mL of USP Bacampicillin Hydrochloride RS in alcohol Sample solution: 2 mg/mL of ampicillin from powdered Tablets diluted in alcohol Developing solvent system: Methylene chloride, chloroform, and alcohol (10:1:1) Spray reagent: 10 mg/mL of ninhydrin in butyl alcohol with 1 mL of pyridine in each 100 mL of solution Chromatographic system (See Chromatography 621.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Analysis Samples: Sample solution and Standard solution Apply two 5-L portions of the Sample solution 4.0 cm apart on a suitable thin-layer chromatographic plate. After the spots dry, apply two 5-L portions of the Standard solution, one midway between the Sample solution spots and the other to one of the Sample solution spots. Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths the length of the plate. Spray the plate with the Spray reagent, and heat at 100 for 10 min. Acceptance criteria: Bacampicillin appears as a purple spot, and the RF values of the spots from the Sample solution and from the combined Sample solution and Standard solution, respectively, correspond to the RF value of the spot obtained from the Standard solution.
USP 32
ASSAY PROCEDURE Solution A: To 0.02 M dibasic sodium phosphate, add portions of 0.02 M monobasic sodium phosphate, until a pH 6.8 0.05 is reached. Mobile phase: Acetonitrile and Solution A (1:1) Standard solution: 0.8 mg/mL of USP Bacampicillin Hydrochloride RS. [NOTESonicate for 20 min to achieve complete dissolution. Pass through a filter of 0.5-m or finer porosity.] Sample solution: Transfer a portion of finely powdered Tablets (NLT 20 Tablets), equivalent to 56 mg of C16H19N3O4S to a 100-mL volumetric flask, add 90 mL of water, and sonicate for 20 min. Dilute to volume with water, and pass through a filter of 0.5-m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3000 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19N3O4S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bacampicillin Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of anhydrous ampicillin, Mr1 349.41 = molecular weight of bacampicillin hydrochloride, Mr2 501.99 Acceptance criteria: 90.0%125.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 30 min Standard solution: 0.3 mg/mL of USP Ampicillin RS in water Analysis: Determine the amounts of C16H19N3O4S dissolved as directed in AntibioticsHydroxylamine Assay, Automated Methods of Analysis 16, Procedure. Tolerances: NLT 85% (Q) of the labeled amount of C16H19N3O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 2.5%
Official Monographs / Bacitracin 3
Bacitracin
(Comment on this Monograph)id=m6830=Bacitracin=BMonos.pdf)
Bacitracin [1405-87-4]. DEFINITION Bacitracin is a polypeptide produced by the growth of an organism of the licheniformis group of Bacillus subtilis (Fam. Bacillacaea). It has a potency of NLT 40 Bacitracin Units/mg. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: Meets the requirements ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81: Proceed as directed in the chapter for Bacitracin. Acceptance criteria: NLT 40 Bacitracin Units/mg SPECIFIC TESTS PH 791: 5.57.5, in a solution containing 10,000 Bacitracin Units/mL LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 5.0% of its weight. STERILITY TESTS 71: Where the label states that the Bacitracin is sterile, it meets the requirements. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Bacitracin is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a cool place. LABELING: Where it is packaged for prescription compounding, label it to indicate that it is not sterile and that the potency cannot be assured for longer than 60 days after opening, and to state the number of Bacitracin Units per milligram. Where it is intended for use in preparing injectable or other sterile dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable or other sterile dosage forms. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS USP Endotoxin RS
Bacitracin for Injection

(Comment on this Monograph)id=m6840=Bacitracin for Injection=B-Monos.pdf) DEFINITION Bacitracin for Injection has a potency of NLT 50 Bacitracin Units /mg. It contains NLT 90.0% and NMT 115.0% of the labeled amount of bacitracin.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Ampicillin RS USP Bacampicillin Hydrochloride RS
Bacitracin / Official Monographs

IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: Meets the requirements
USP 32
IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: Meets the requirements ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 Sample solution 1: Constitute one container of Bacitracin for Injection as directed in the labeling. Using a suitable hypodermic needle and syringe, withdraw the contents of the container, and dilute quantitatively with Buffer No. 1 to obtain a solution containing about 100 Bacitracin Units/mL. Sample solution 2 (where the label states the number of Bacitracin Units in a given volume of constituted solution): Constitute one container of Bacitracin for Injection as directed in the labeling. Dilute a volume of the constituted solution with Buffer No. 1 to obtain a solution containing 100 Bacitracin Units/mL. Analysis Samples: Sample solution 1 and Sample solution 2 Proceed as directed. Add sufficient 0.01 N hydrochloric acid to the Sample solution so that the amount of hydrochloric acid in the Test Dilution will be the same as in the median dose level of the Standard, and dilute with Buffer No. 1 to obtain a Test Dilution having a bacitracin concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 Sample: Use a portion of Ointment shaken with about 50 mL of ether in a separator, and extracted with four 20-mL portions of Buffer No. 1. Combine the buffer extracts, and dilute with Buffer No. 1 to an appropriate volume to obtain a stock solution. Add sufficient 0.01 N hydrochloric acid to a measured portion of the stock solution so that the amount of hydrochloric acid in the Test Dilution will be the same as in the median dose level of the Standard, and quantitatively dilute with Test Dilution having a bacitracin concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%140.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5% [NOTEUse 20 mL of a mixture of toluene and methanol (7:3) in place of methanol in the titration vessel.] MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers containing NMT 60 g, unless labeled solely for hospital use, preferably at controlled room temperature. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 3.0%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid HEAVY METALS, Method II 231: NMT 30 ppm SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements under Injections 1. PH 791: 5.57.5, 10,000 Bacitracin Units/mL LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure of NMT 5 mm of mercury at 60 for 3 h: it loses NMT 5.0% of its weight. INJECTIONS 1: Meets the requirements STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.01 USP Endotoxin Unit/Bacitracin Unit. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids, and store in a cool place. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS USP Endotoxin RS
Bacitracin Ophthalmic Ointment

(Comment on this Monograph)id=m6870=Bacitracin Ophthalmic Ointment=B-Monos.pdf) DEFINITION Bacitracin Ophthalmic Ointment is a sterile preparation of Bacitracin in an anhydrous ointment base. It contains NLT 90.0% and NMT 140.0% of the labeled amount of bacitracin. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: Meets the requirements ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 Sample: Use a portion of Ophthalmic Ointment shaken with 50 mL of ether in a separator, and extracted with four 20-mL portions of Buffer No. 1. Combine the buffer extracts, and dilute with Buffer No. 1 to an appropriate volume to obtain a stock solution. Add sufficient 0.01 N hydrochloric acid to a portion of the stock solution so that the amount of hydrochloric acid in the Test Dilution will be the same as in the median dose level of the Standard, and quantitatively dilute with Test Dilution having a bacitracin concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%140.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5% [NOTEUse 20 mL of a mixture of toluene and methanol (7:3) in place of methanol in the titration vessel.] METAL PARTICLES IN OPTHALMIC OINTMENTS 751: Meets the requirements
Bacitracin Ointment
(Comment on this Monograph)id=m6860=Bacitracin Ointment=B-Monos.pdf) DEFINITION Bacitracin Ointment is Bacitracin in an anhydrous ointment base. It contains NLT 90.0% and NMT 140.0% of the labeled amount of bacitracin. It may contain a suitable anesthetic.
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STERILITY TESTS 71: Meets the requirements

toluene and methanol (7:3) in place of methanol in the titration vessel.] OTHER REQUIREMENTS It meets the requirements for Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers 601, Pressure Test, Minimum Fill, and Leakage Test. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in pressurized containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS USP Polymyxin B Sulfate RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS
Bacitracin and Polymyxin B Sulfate Topical Aerosol

(Comment on this Monograph)id=m6885=Bacitracin and Polymyxin B Sulfate Topical Aerosol=B-Monos.pdf) DEFINITION Bacitracin and Polymyxin B Sulfate Topical Aerosol is a suspension of Bacitracin and Polymyxin B Sulfate in a suitable vehicle, packaged in a pressurized container with a suitable inert propellant. It contains NLT 90.0% and NMT 130.0% of the labeled amounts of bacitracin and polymyxin B. It may contain a suitable local anesthetic. [NOTEPrepare the specimen for the following tests and assays as follows. Maintain the container in the inverted position throughout this procedure. Store the container in a freezer at 70 for 16 to 24 h. Remove the container from the freezer, promptly puncture the container, and allow the propellant to volatilize. Open the container, and mix the contents.] IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: A portion of the contents of one container prepared as directed above and tested as directed under For Creams, Lotions, and Ointments meets the requirements. ASSAY BACITRACIN (See AntibioticsMicrobial Assays 81.) Sample: A portion of the contents of one container, prepared as directed, equivalent to about 500 USP Bacitracin Units Analysis: Transfer to a suitable separator containing 50 mL of ether, and extract with three 25-mL portions of Buffer No. 1. Combine the buffer extracts in a 100-mL volumetric flask, dilute with Buffer No. 1 to volume, and mix. Add sufficient 0.01 N hydrochloric acid to this solution so that the amount of hydrochloric acid in the Test solution will be the same as in the median dose level of the Standard, and quantitatively dilute with Buffer No. 1 to obtain a Test Dilution having a bacitracin concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%130.0% POLYMYXIN B (See AntibioticsMicrobial Assays 81.) Sample: A portion of the contents of one container, prepared as directed, equivalent to 5000 USP Polymyxin B Units Analysis: Transfer to a suitable separator containing 50 mL of ether, and extract with three 25-mL portions of Buffer No. 6. Combine the buffer extracts in a 100-mL volumetric flask, dilute with Buffer No. 6 to volume, and mix. Dilute this solution, quantitatively and stepwise, with Buffer No. 6 to obtain a Test Dilution having a polymyxin B concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%130.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5% [NOTEUse a portion of the contents of one container, prepared as directed above, and 20 mL of a mixture of
Soluble Bacitracin Methylene Disalicylate

(Comment on this Monograph)id=m6890=Soluble Bacitracin Methylene Disalicylate=B-Monos.pdf) DEFINITION Soluble Bacitracin Methylene Disalicylate is a mixture of Bacitracin Methylene Disalicylate and Sodium Bicarbonate. It has a potency of NLT 8 Bacitracin Units/mg, calculated on the dried basis. ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 Sample stock solution: Transfer a sample to a high-speed glass blender jar, add 99.0 mL of a 20 mg/mL sodium bicarbonate solution and 1.0 mL of polysorbate 80, and blend for 3 min. Add a sufficient volume of 0.01 N hydrochloric acid so that the amount of hydrochloric acid will be the same as in the median dose level of the Standard. Sample solution: Dilute the Sample stock solution with Buffer No. 1 (see Phosphate Buffers and Other Solutions) to obtain a concentration of bacitracin assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed for Bacitracin in Antibiotics Microbial Assays 81. Acceptance criteria: NLT 8 Bacitracin Units/mg SPECIFIC TESTS PH 791: 8.09.5, in a solution of 25 mg/mL LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 8.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS
Bacitracin Methylene Disalicylate Soluble Powder

(Comment on this Monograph)id=m6894=Bacitracin Methylene Disalicylate Soluble Powder=B-Monos.pdf) DEFINITION Bacitracin Methylene Disalicylate Soluble Powder contains NLT 90.0% and NMT 120.0% of the labeled amount of bacitracin.
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Standard stock solution: Equivalent to 10 mg/mL of zinc (from zinc oxide) in 1 N hydrochloric acid [NOTEWarm to dissolve then allow to cool before diluting to volume.] Standard solutions: 0.5, 1.5, and 2.5 g/mL of zinc in 0.001 N hydrochloric acid, from Standard stock solution Sample stock solution: 2 mg/mL of Bacitracin Zinc in 0.01 N hydrochloric acid Sample solution: 0.02 mg/mL in 0.001 N hydrochloric acid, from Sample stock solution Blank: 0.001 N hydrochloric acid Spectrometric conditions (See Spectrophotometry and Light-Scattering 851). Mode: Atomic absorption spectrophotometry Detector: Zinc hollow-cathode lamp and an airacetylene flame Analytical wavelength: Zinc resonance line of 213.8 nm Analysis Samples: Standard solutions, Sample solution, and Blank. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of zinc, and draw the straight line best fitting the three plotted points and determine the concentration, in g/mL, of zinc in the Sample solution. Calculate the content of zinc, as a percentage, in the portion of Bacitracin Zinc taken: Result = C/Cu 100 = concentration of zinc in the Sample solution obtained from the curve (g/mL) Cu = concentration of Bacitracin Zinc in the Sample solution (g/mL) Acceptance criteria: 4.0%-6.0% of Zinc, calculated on the dried basis COMPOSITION Solution A: Dissolve 34.8 g of potassium phosphate, dibasic, in 1 L of water. Adjust with 27.2 g of potassium phosphate, monobasic, dissolved in 1 L of water, to a pH of 6.0. Mobile phase: Mixture of methanol, acetonitrile, Solution A, and water (26:2:5:15) Mix well, and degas. Diluent: Dissolve 40 g of sodium edentate in 1 L of water. Adjust with dilute sodium hydroxide to a pH of 7.0. System suitability solution: 2.0 mg/mL of USP Bacitracin Zinc RS in Diluent Reporting threshold solution: Dilute quantitatively, with water, a suitable volume of the System suitability solution to obtain a solution with a known concentration of 0.01 mg/mL. Peak identification solution: Dissolve a quantity of USP Bacitracin Zinc RS in a suitable volume of Diluent to obtain a 2.0 mg/mL solution. Heat in a boiling water bath for 30 min. Cool to room temperature. Sample solution: 2.0 mg/mL of Bacitracin Zinc in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 250-mm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: Peak identification solution and System suitability solution C
ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 Sample stock solution: Transfer a sample to a high-speed glass blender jar. Add 99.0 mL of sodium bicarbonate solution (1 in 50) and 1.0 mL of polysorbate 80, and blend for about 3 min. Add a sufficient volume of 0.01 N hydrochloric acid to a measured volume of Sample solution so that the amount of hydrochloric acid will be the same as in the median dose level of the Standard. Sample solution: Dilute the Sample stock solution with Buffer No. 1 (see Phosphate Buffers and Other Solutions) to obtain a concentration of bacitracin assumed to be equal to the median dose level of the Standard. SPECIFIC TESTS PH 791: 8.09.5, 50 mg/mL LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 8.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that it is for veterinary use only. Label it to state the content of bacitracin in terms of grams per pound, each gram of bacitracin being equivalent to 42,000 Bacitracin Units. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS
Bacitracin Zinc
(Comment on this Monograph)id=m6910=Bacitracin Zinc=BMonos.pdf) Bacitracins zinc complex [1405-89-6]. DEFINITION Bacitracin Zinc is the zinc complex of bacitracin, which consists of a mixture of antimicrobial polypeptides, the main components being bacitracins A, B1, B2, and B3. It has a potency of NLT 65 Bacitracin Units/mg, calculated on the dried basis. It contains NLT 4.0% and NMT 6.0% of zinc (Zn), calculated on the dried basis. IDENTIFICATION A. Thin-Layer Chromatographic Identification Test 201BNP: Meets the requirements B. Meets the requirements of the liquid chromatographic procedure in the test for Composition ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 : NLT 65 Bacitracin Units/mg, calculated on the dried basis SPECIFIC TESTS PH 791: 6.07.5, in a (saturated) solution 100 mg/mL LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at 60 for 3 h: it loses NMT 5.0% of its weight. STERILITY TESTS 71: Where the label states that it is sterile, it meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration, except to add 20g of edetate disodium to Fluid A. ZINC CONTENT [NOTEThe Standard solutions and the Sample solution may be quantitatively diluted with 0.001 N hydrochloric acid, if necessary, to obtain solutions of suitable concentrations, adaptable to the linear or working range of the instrument.]
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Suitability requirements Peak-to-valley ratio: NLT 1.2
Impurity Table 1 Relative Retention Time (approximate) 0.5 0.6 0.6 0.7 0.7 0.8 1.0 2.4

Acceptance criteria: The sum of bacitracin A, B1, B2, and B3 is NLT 70.0% of the Total area. Limit of Early Eluting Peptides Calculate the percentage of all peaks eluting before the peak due to bacitracin B1: Result = (rPreB1/Total area) 100 rPreB1 = sum of the responses of all peaks eluting before the peak for bacitracin B1 Acceptance criteria: The limit of early eluting peptides is NMT 20.0%. Limit of Bacitracin F Calculate the percentage of bacitracin F: Result = (rF/rA) 100 rF = response of bacitracin F from the Sample solution rA = response of bacitracin A from the Sample solution Acceptance criteria: The limit of bacitracin F, a known impurity, is NMT 6.0%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a cool place. LABELING: Label it to indicate that it is to be used in the manufacture of nonparenteral drugs only. Where it is packaged for prescription compounding, label it to indicate that it is not sterile and that the potency cannot be assured for longer than 60 days after opening, and to state the number of Bacitracin Units/milligram. Where it is intended for use in preparing sterile dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of sterile dosage forms. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS
Name Bacitracin C1 Bacitracin C2 Bacitracin C3 Bacitracin B1 Bacitracin B2 Bacitracin B3 Bacitracin A Bacitracin F
Chromatograph the Peak identification solution (detector set at 300 nm) and then the System suitability solution (detector set at 254 nm). Identify the location of bacitracin F, which is a known impurity, using the relative retention time shown in Impurity Table 1. Identify the peaks of the most active components of bacitracin (bacitracins A, B1, B2, and B3), early eluting peptides (those eluting before the peak due to bacitracin B1), and the impurity, bacitracin F, using the relative retention values in Impurity Table 1. Calculate the peak-to-valley ratio: Result = HP/HV HP HV = height above the baseline of the peak due to bacitracin B1 = height above the baseline of the lowest point of the curve separating the bacitracin B1 peak from the peak due to bacitracin B2
Analysis Sample: Diluent, Sample solution, and Reporting threshold solution. Record the chromatograms for about three times the retention time of bacitracin A. Identify the peaks using the relative retention times shown in Impurity Table 1. [NOTEDisregard any peak in the Sample solution having an area less than the area of the bacitracin A peak in the Reporting threshold solution; disregard any peak observed in the Diluent.] [NOTEThe total area in the following calculations is defined as the area of all peaks except the reporting threshold.] Content of Bacitracin A Calculate the percentage of bacitracin A: Result = (rA/Total area) 100 = area response from bacitracin A rA Acceptance criteria: Bacitracin A content is NLT 40.0% of the Total area. Content of Active Bacitracin Calculate the percentage of active bacitracin (bacitracin A, B1, B2, and B3): Result = (rS/Total area) 100 rS = rA, rB1, rB2, and rB3 area responses from bacitracin A, B1, B2, and B3, respectively
Bacitracin Zinc Ointment

(Comment on this Monograph)id=m6915=Bacitracin Zinc Ointment=B-Monos.pdf) DEFINITION Bacitracin Zinc Ointment is Bacitracin Zinc in an anhydrous ointment base. It contains NLT 90.0% and NMT 140.0% of the labeled amount of bacitracin. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: Meets the requirements ASSAY PROCEDURE (See AntibioticsMicrobial Assays 81.) Standard solution: For each test dilution of the Standard, add additional hydrochloric acid to obtain the same concentration of hydrochloric acid as in the Sample solution. Sample stock solution: Weigh a portion of Ointment and shake with about 50 mL of ether in a separator. Extract with four 20-mL portions of 0.01 N hydrochloric acid. Combine the acid extracts, and dilute with 0.01 N hydrochloric acid to an appropriate volume. Sample solution: Dilute the Sample stock solution quantitatively and stepwise with Buffer No. 1 to obtain a concentration assumed to be equal to the median dose level of the Standard solution.

Analysis: Proceed as directed. Acceptance criteria: NLT 90.0% and NMT 140.0%
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best fitting the three plotted points. From the graph so obtained, determine the concentration, in g/mL, of zinc in the Sample solution. Calculate the zinc content, in g, in relation to each 42,000 Bacitracin Units in the specimen taken: Result = 280,000C/(W A) = concentration of zinc in the Sample solution (g/mL) W = weight of the Powder taken (mg) A = bacitracin content in the Powder (Bacitracin Units/g) Acceptance criteria: NMT 2.0 g for each 42,000 Bacitracin Units SPECIFIC TESTS LOSS ON DRYING 731 Sample: 100 mg Analysis: Dry the Sample in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h. Acceptance criteria: The sample loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that it is for veterinary use only. Label it to state the content of bacitracin in terms of g/lb, each g of bacitracin being equivalent to 42,000 Bacitracin Units. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS C
SPECIFIC TESTS WATER DETERMINATION, Method I 921 Analysis: Proceed as directed, except use 20 mL of a mixture of toluene and methanol (7:3) in place of methanol in the titration vessel. Acceptance criteria: NMT 0.5% MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers containing NMT 60 g, unless labeled solely for hospital use, preferably at controlled room temperature. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS
Bacitracin Zinc Soluble Powder

(Comment on this Monograph)id=m6917=Bacitracin Zinc Soluble Powder=B-Monos.pdf) DEFINITION Bacitracin Zinc Soluble Powder is a mixture of Bacitracin Zinc and zinc proteinates. It contains NLT 90.0% and NMT 120.0% of the labeled amount of bacitracin. ASSAY PROCEDURE (See AntibioticsMicrobial Assays 81.) Standard solution: For each test dilution of the Standard, add additional hydrochloric acid to each to obtain the same concentration of hydrochloric acid as in the Sample solution. Sample stock solution: Equivalent to 100 Bacitracin Units/mL in 0.01 N hydrochloric acid Sample solution: Dilute the Sample stock solution stepwise with Buffer No. 1 to obtain a concentration assumed to be equal to the median dose level of the Standard solution. Analysis: Proceed as directed. Acceptance criteria: NLT 90.0% and NMT 120.0% OTHER COMPONENTS ZINC CONTENT [NOTEThe Standard solutions and the Sample solution may be diluted with 0.001 N hydrochloric acid, if necessary, to obtain solutions of suitable concentrations, adaptable to the linear or working range of the instrument.] Standard stock solution: Transfer 3.11 g of zinc oxide to a 250-mL volumetric flask, add 80 mL of 1 N hydrochloric acid, warm to dissolve, cool, and dilute with water to volume. This solution contains 10 mg/mL of zinc. Standard solutions: 0.5 g/mL, 1.5 g/mL, and 2.5 g/mL of zinc in 0.001 N hydrochloric acid: from the Standard stock solution Sample solution: Transfer an amount of Powder equivalent to 200 mg of Bacitracin Zinc to a 100-mL volumetric flask. Dissolve in and dilute with 0.01 N hydrochloric acid to volume. Dilute 2 mL of this solution with 0.001 N hydrochloric acid to 200 mL. Blank: 0.001 N hydrochloric acid Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometry Detector: Zinc hollow-cathode lamp and an air-acetylene flame Analytical wavelength: Zinc resonance line, 213.8 nm Analysis: Sample: Standard solutions, Sample solution, and Blank. Plot the absorbances of the Standard solutions versus concentration, in g/mL of zinc, and draw the straight line
Bacitracin Zinc and Polymyxin B Sulfate Ointment

(Comment on this Monograph)id=m6926=Bacitracin Zinc and Polymyxin B Sulfate Ointment=B-Monos.pdf) DEFINITION Bacitracin Zinc and Polymyxin B Sulfate Ointment contains the equivalent of NLT 90.0% and NMT 130.0% of the labeled amounts of bacitracin and polymyxin B. It may contain a suitable local anesthetic. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: Meets the requirements ASSAY BACITRACIN (See AntibioticsMicrobial Assays 81.) Analysis: Shake a portion of Ointment with about 50 mL of ether in a separator, and extract with four 20-mL portions of 0.01 N hydrochloric acid. Combine the acid extracts, and dilute with 0.01 N hydrochloric acid to an appropriate volume to obtain a Sample solution. Dilute this Sample solution quantitatively and stepwise with Buffer No. 1 to obtain a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard, adding additional hydrochloric acid to each Dilute solution of the Standard to obtain the same concentration of hydrochloric acid as in the Test Dilution. POLYMYXIN B (See AntibioticsMicrobial Assays 81.) Analysis: Transfer a portion of Ointment to a suitable separator containing 50 mL of ether, and extract with four 20-mL portions of Buffer No. 6. Combine the aqueous extracts, and dilute with Buffer No. 6 to an appropriate volume to obtain a Sample solution. Dilute this Sample solution quantitatively and stepwise with Buffer No. 6 to
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obtain a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%130.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS USP Polymyxin B Sulfate RS
Official Monographs / Baclofen 9

STERILITY TESTS 71 Analysis: Test as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. Acceptance criteria: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Bacitracin Zinc RS USP Polymyxin B Sulfate RS
Baclofen
(Comment on this Monograph)id=m6940=Baclofen=BMonos.pdf)
Bacitracin Zinc and Polymyxin B Sulfate Ophthalmic Ointment

(Comment on this Monograph)id=m6927=Bacitracin Zinc and Polymyxin B Sulfate Ophthalmic Ointment=B-Monos.pdf) DEFINITION Bacitracin Zinc and Polymyxin B Sulfate Ophthalmic Ointment contains the equivalent of NLT 90.0% and NMT 130.0% of the labeled amounts of bacitracin and polymyxin B. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201BNP: Meets the requirements ASSAY BACITRACIN (See AntibioticsMicrobial Assays 81.) Standard solution: For each test dilution of the Standard, add additional hydrochloric acid to each to obtain the same concentration of hydrochloric acid as in the Sample solution. Sample stock solution: Weigh a portion of Ophthalmic Ointment and shake with about 50 mL of ether in a separator. Extract with four 20-mL portions of 0.01 N hydrochloric acid. Combine the acid extracts, and dilute with 0.01 N hydrochloric acid to an appropriate volume. Sample solution: Dilute the Sample stock solution with Buffer No. 1 to obtain a concentration assumed to be equal to the median dose level of the Standard solution. Analysis: Proceed as directed. Acceptance criteria: NLT 90.0% and NMT 130.0% POLYMYXIN B (See AntibioticsMicrobial Assays 81.) Sample stock solution: Weigh a portion of Ophthalmic Ointment and shake with 50 mL of ether in a separator. Extract with four 20-mL portions of Buffer No. 6. Combine the aqueous extracts, and dilute with Buffer No. 6 to an appropriate volume. Sample solution: Dilute the Sample stock solution with Buffer No. 6 to obtain a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed. Acceptance criteria: NLT 90.0% and NMT 130.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921 Analysis: Proceed as directed, except use 20 mL of a mixture of toluene and methanol (7:3) in place of methanol in the titration vessel. Acceptance criteria: NMT 0.5% MINIMUM FILL 755: Meets the requirements METAL PARTICLES IN OPHTHALMIC OINTMENTS 751: Meets the requirements C10H12ClNO2 213.66 Butanoic acid, 4-amino-3-(4-chlorophenyl)-; -(Aminomethyl)-p-chlorohydrocinnamic acid [1134-47-0]. DEFINITION Baclofen contains NLT 99.0% and NMT 101.0% of C10H12ClNO2, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197M ASSAY PROCEDURE Sample solution: Dissolve 200 mg of Baclofen in a suitable volume of glacial acetic acid, sufficient to immerse the electrodes. Analysis: Titrate the Sample solution with 0.1 N perchloric acid VS, using a calomel electrode containing a saturated solution of lithium chloride in glacial acetic acid. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 21.37 mg of C10H12ClNO2. Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.3% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE [NOTEAvoid contact with o-tolidine when performing this test, and conduct the test in a well-ventilated hood.] Diluent: Alcohol and glacial acetic acid (4:1) Standard stock solution: 0.1 mg/mL of USP Baclofen RS in Diluent Standard solution A: 0.05 mg/mL of USP Baclofen RS in Diluent, from the Standard stock solution Standard solution B: 0.03 mg/mL of USP Baclofen RS in Diluent, from the Standard stock solution Identification solution: 0.1 mg/mL USP Baclofen Related Compound A RS in Diluent Sample solution: 10 mg/mL of Baclofen in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Adsorbent: 0.25-mm layer of chromatographic silica gel mixture
10
Baclofen / Official Monographs

Application volume: 10 L Developing solvent system: Butyl alcohol, glacial acetic acid, and water (4:1:1) Spray reagent: Dissolve 200 mg of o-tolidine in 2 mL of glacial acetic acid with the aid of a hot water bath. Dilute to 100 mL with water. Mix equal volumes of o-tolidine solution and 0.83% potassium iodide solution (1:1). Analysis Samples: Standard solution A, Standard solution B, Identification solution, and Sample solution Proceed as directed. Remove the plate from the developing chamber, and dry under a current of warm air. Transfer the dry plate to another chromatographic chamber containing a beaker with 1 g of potassium permanganate. Add 10 mL of 40% hydrochloric acid to the beaker. Cover the chamber, and allow the chamber to become saturated with chlorine gas. Expose the plate to the chlorine gas for 8 min. Remove the plate and expose to the air for 2 min, then spray with freshly prepared Spray reagent. Acceptance criteria: The intensity of the secondary spot from the Sample solution, corresponding in RF value to the principal spot from the Identification solution, is NMT that of the principal spot from the Identification solution (1.0%), and no other secondary spot from the Sample solution is greater in intensity than the principal spot from Standard solution A (0.5%). The sum of all secondary spots from the Sample solution is NMT 2.0%.
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Adjust with phosphoric acid to a pH of 3.5. Standard stock solution: 1.0 mg/mL of USP Baclofen RS in water Standard solution: 5 g/mL of USP Baclofen RS in water, from the Standard stock solution Sample solution: Shake thoroughly by hand each bottle of Oral Suspension. Pipet 0.5 mL of Oral Suspension from each bottle to a 500-mL volumetric flask, dilute with water to volume to obtain a concentration of 5 g/mL, and pass through a 0.22-m polyvinylidenefluoride (PVDF) filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 15 L System suitability Sample: Standard solution [NOTEThe retention time of baclofen is about 5.5 min.] Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H12ClNO2 in the volume of Oral Suspension: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Baclofen RS in the Standard solution (g/mL) CU = nominal concentration of baclofen in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.25.2 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store in a cold place. LABELING: Label it to state that it is to be well shaken, and to state the beyond-use date. BEYOND-USE DATE: 35 days after the day on which it was compounded USP REFERENCE STANDARDS 11 USP Baclofen RS rU rS CS
NMT 3.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Baclofen RS USP Baclofen Related Compound A RS
Baclofen Oral Suspension

(Comment on this Monograph)id=m1722=Baclofen Oral Suspension=B-Monos.pdf) DEFINITION Baclofen Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of baclofen (C10 H12 ClNO2). Prepare Baclofen Oral Suspension 5 mg/mL as follows (see Pharmaceutical CompoundingNonsterile Solutions 795):
Baclofen Vehicle: a mixture of Vehicle for Oral Solution(regular or sugar-free), NF, and Vehicle for Oral Suspension, NF (1:1) To make 500 mg A sufficient quantity
Baclofen Tablets
(Comment on this Monograph)id=m6970=Baclofen Tablets=BMonos.pdf) DEFINITION Baclofen Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C10H12ClNO2. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Equivalent to 5 mg/mL of baclofen from powdered Tablets in dehydrated alcohol and glacial acetic acid (4:1) [NOTEShake for 30 min and centrifuge.] Standard solution: 5 mg/mL of USP Baclofen RS in dehydrated alcohol and glacial acetic acid (4:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
100 mL
If using Baclofen Tablets, place the Tablets in a suitable mortar and comminute the Tablets to a fine powder, or add Baclofen powder. Add 5 mL of the Vehicle to wet the powder, and triturate the powder to form a fine paste. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer, stepwise and quantitatively, the contents of the mortar to a calibrated bottle. Add sufficient Vehicle to bring to final volume, and mix well. ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.05 M monobasic sodium phosphate (1:4)
USP 32
Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Developing solvent system: Butyl alcohol, glacial acetic acid, and water (4:1:1) Spray reagent: 0.4 g of ninhydrin in 95 mL of butyl alcohol mixed with 5 mL of dilute glacial acetic acid (1 in 10) Analysis: Proceed as directed. Develop the chromatogram until the solvent front has moved about three-fourths the length of the plate. Remove the plate from the developing chamber, and dry under a current of warm air. Spray the plate with the Spray reagent until the plate is slightly wet. Place the plate in an oven maintained at 100 for 10 min. Acceptance criteria: The RF value of the principal orangered spot of the Sample solution corresponds to that of the Standard solution. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol, glacial acetic acid, and water (15:2:33) Mobile phase: 0.3 N acetic acid, Methanol, and 0.36 N sodium 1-pentanesulfonate (55:44:2) Standard solution: 4 mg/mL of USP Baclofen RS in Diluent Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer an amount equivalent to 40 mg to a 50-mL flask. Add 10.0 mL of Diluent to the flask. Sonicate to disperse, and shake by mechanical means for 30 min. Centrifuge a portion of this solution for 5 min, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 0.6 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample Solution [NOTESample solution to elute at NLT three times the retention time of baclofen.] Result = (rU/rS) (CS/CU) 100 = peak area response of the Sample solution = peak area response of the Standard solution = concentration of USP Baclofen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution Cu (mg/mL) Acceptance criteria: 90.0%110.0% rU rS Cs PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: 0.01 N hydrochloric acid; for NMT 10 mg of drug, 500 mL for more than 10 mg of drug, 1000 mL Apparatus 2: 50 rpm Time: 30 min Mobile phase: Proceed as directed in the Assay. Standard solution: USP Baclofen RS in Medium Sample solution: Sample per Dissolution 711 Chromatographic system (See Chromatography 621, System Suitability.)
Official Monographs / Bandage 11

Mode: LC Detector: UV 265 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 0.6 mL/min Injection size: 190 L Analysis Samples: Sample solution and Standard solution Calculate the quantity of C10H12ClNO2 dissolved. Tolerances: NLT 75% (Q) of the labeled amount of C10H12ClNO2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Diluent: Proceed as directed in the Assay. Mobile phase: Proceed as directed in the Assay. Standard stock solution: 1 mg/mL of USP Baclofen Related Compound A RS in methanol Standard solution: 0.16 mg/mL of USP Baclofen Related Compound A in Diluent from the Standard stock solution Sample solution: Proceed as directed in the Assay. Chromatographic system and System suitability: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of baclofen related compound A in the portion of the Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak area response of the Sample solution = peak area response of the Standard solution = concentration of USP Baclofen Related Compound A RS in the Standard solution (mg/mL) = nominal concentration of baclofen in the Sample CU solution (mg/mL) Acceptance criteria: NMT 4.0% of baclofen related compound A rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Baclofen RS USP Baclofen Related Compound A RS
Adhesive Bandage
(Comment on this Monograph)id=m7020=Adhesive Bandage=B-Monos.pdf) DEFINITION Adhesive Bandage consists of a compress of four layers of Type I Absorbent Gauze or other suitable material affixed to a film or fabric coated with a pressure-sensitive adhesive substance. It is sterile. The compress may contain a suitable antimicrobial agent and may contain one or more suitable colors. The adhesive surface is protected by a suitable removable covering. SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Package Adhesive Bandage that does not exceed 15 cm (6 inches) in width individually in such manner that sterility is maintained until the individual
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Bandage / Official Monographs
USP 32
WIDTH: Measure width at each of the 5 points selected for the determination of the thread count: the average of 5 measurements is NMT 1.6 mm (1/16 inch) less than the labeled width of the Bandage. LENGTH: Measure the length of the unrolled Gauze Bandage, smoothed without tension, along the center line of the Gauze Bandage: the length is NLT 98.0% of the labeled length of the Bandage. WEIGHT: Weigh the entire Bandage: the calculated weight in g/0.894 m2 (1 linear yard Type I gauze), using the measurements obtained as described under Width and Length is NLT 39.2 g. ABSORBENCY: Hold a rolled Gauze Bandage horizontal to and almost in contact with the surface of water at 25, and allow to drop lightly upon the water: complete submersion takes place in NMT 30 s. IGNITED RESIDUE, ACID OR ALKALI, AND DEXTRIN OR STARCH, IN WATER EXTRACT Sample solution: Place 20 0.1 g of Gauze Bandage in 500 mL of water, and boil the mixture for 15 min, adding boiling water as necessary to maintain the original volume. Pour the water through a funnel into a 1000-mL volumetric flask, transfer the Absorbent Gauze to the funnel, press out the excess water with a glass rod, and wash it with two 250-mL portions of boiling water, pressing the gauze after each washing. Cool the combined washings, dilute to volume, and mix. Analysis 1 (Ignited Residue): Evaporate 400 mL of the Sample solution, filtering if necessary, in a suitable dish on a steam bath, and dry the residue at 105 to constant weight. Ignite the dried residue in a muffle furnace at a dull-red heat to constant weight. Acceptance criteria 1: The weight of the ignited residue does not exceed an amount, in mg, calculated as follows: Result = 20 0.14C = corrected percentage of cotton (13 mg maximum, or 0.16%) Analysis 2 (Acid or Alkali): To separate 200-mL portions of the Sample solution, add 3 drops of phenolphthalein TS and 1 drop of methyl orange TS, respectively. Acceptance criteria 2: No pink color develops in either portion. Analysis 3 (Dextrin or Starch): To a 200-mL portion of the Sample solution, add 1 drop of iodine TS. Acceptance criteria 3: No red, violet, or blue color develops. ALCOHOL-SOLUBLE DYES: Pack 10 g of Gauze Bandage in a narrow percolator, and extract slowly with alcohol until the percolate measures 50 mL: when observed downward in a column 20 cm in depth, the percolate may show a yellowish color, but neither a blue nor a green tint. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Gauze Bandage that has been rendered sterile is so packaged that the sterility of the contents is maintained until the package is opened for use. LABELING: The width and length of the Bandage, the number of pieces contained, and the name of the manufacturer, packer, or distributor, are stated on the package. The designation non-sterilized or not sterilized appears prominently on the package unless the Gauze Bandage has been rendered sterile, in which case it may be labeled to indicate that it is sterile and that the contents may not be sterile if the package bears evidence of damage or if the package has been previously opened. C
package is opened. Package individual packages in a second protective container. LABELING: The label of the second protective container bears a statement that the contents may not be sterile if the individual package has been damaged or previously opened, and it bears the names of any added antimicrobial agents. Each individual package is labeled to indicate the dimensions of the compress and the name of the manufacturer, packer, or distributor, and each protective container indicates also the address of the manufacturer, packer, or distributor.
Gauze Bandage
(Comment on this Monograph)id=m7030=Gauze Bandage=BMonos.pdf) DEFINITION Gauze Bandage is Type I Absorbent Gauze. Its length is NLT 98.0% of that declared on the label, and its average width is NMT 1.6 mm less than the declared width. It contains no dye or other additives. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION Sample: Place 5 g in a suitable dish, and moisten with 2 N sulfuric acid. Analysis: Gently heat the Sample mixture until it is charred, then ignite more strongly until the carbon is completely consumed. Acceptance criteria: The weight of the residue corresponds to NMT the percentage of the weight of the Gauze, calculated as follows: Result = 0.002C + 0.015(100 C) = corrected percentage of cotton (0.89% maximum) Organic Impurities FATTY MATTER Sample solution: Pack 10 0.01 g of Gauze Bandage in a continuous-extraction thimble with a tared flask, and extract with ether for 5 h, adjusting the rate so that the ether siphons NLT four times/h. [NOTEThe ether extract in the flask shows no trace of blue, green, or brownish color.] Analysis: Evaporate the Sample solution extract to dryness, and dry at 105 to constant weight. Acceptance criteria: The weight of the residue does not exceed an amount, in mg, calculated as follows: Result = 0.4C + 30 C = corrected percentage of cotton (70 mg maximum, or 0.7%) C
SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements THREAD COUNT: Count the number of warp and filling threads in areas of 1.27 cm (1/2 inch) square at 5 points evenly spread along the center line of the Bandage, no point being within 30.5 cm (12 inches) of either end of the Bandage, and calculate the average number of threads/2.54 cm (1 inch) in each direction. A variation of NMT 3 threads/inch is allowed in either warp or filling, provided that the combined variations do not exceed 5 threads/square inch. [NOTEBefore determining the thread count, dimensions, and weight, hold the Bandage, unrolled, for NLT 4 h in a standard atmosphere of 65 2% relative humidity at 21 1.1C (70 2F).]
USP 32
Official Monographs / Barium 13

Acceptance criteria: The increase in weight is NLT 19.0% of the weight of Barium Hydroxide Lime used for the test. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: If an indicator has been added, the name and color change of such indicator are stated on the container label. The container label indicates also the mesh size in terms of standard-mesh sieve sizes (see Powder Fineness 811).
Barium Hydroxide Lime

(Comment on this Monograph)id=m7080=Barium Hydroxide Lime=B-Monos.pdf) DEFINITION Barium Hydroxide Lime is a mixture of barium hydroxide octahydrate and Calcium Hydroxide. It may also contain Potassium Hydroxide and may contain an indicator that is inert toward anesthetic gases such as Ether, Cyclopropane, and Nitrous Oxide and that changes color when the Barium Hydroxide Lime no longer can absorb carbon dioxide. [CAUTIONBecause Barium Hydroxide Lime contains a soluble form of barium, it is toxic if swallowed.] IDENTIFICATION A. PROCEDURE Place a granule of it on a piece of moistened red litmus paper: the paper turns blue immediately. B. IDENTIFICATION TESTSGENERAL, Barium, Calcium, and Potassium 191: A 100 mg/mL solution in 6 N acetic acid responds to the tests for Barium and for Calcium, and it may respond also to the flame test for Potassium. SPECIFIC TESTS PARTICLE SIZE DISTRIBUTION ESTIMATION BY ANALYTICAL SIEVING 786: Screen 100 g for 5 min as directed in the chapter, using a mechanical shaker. It passes completely through a No. 2 standard-mesh sieve, and NMT 2.0% passes through a No. 40 standard-mesh sieve. NMT 7.0% is retained on the coarse-mesh sieve, and NMT 15.0% passes through the finemesh sieve designated on the label. LOSS ON DRYING 731: Weigh about 10 g in a tared weighing bottle, and dry at 105 for 2 h: it loses between 11.0% and 16.0% of its weight. HARDNESS Sample: 200 g Analysis: Screen the Sample on a mechanical sieve shaker (see Particle Size Distribution Estimation by Analytical Sieving 786) having a frequency of oscillation of 285 3 cycles/min, for 3 min, to remove granules coarser than 4mesh and finer than 8-mesh. Weigh 50 g of the granules retained on the screen, and place them in a hardness pan of the following description: the hardness pan has a diameter of 200 mm and a concave brass bottom, and the bottom of the pan is 7.9 mm thick at the circumference and 3.2 mm thick at the center and has an inside spherical radius of curvature of 109 cm. Add 15 steel balls of 7.9-mm diameter, and shake on a mechanical sieve shaker for 30 min. Remove the steel balls, brush the contents of the hardness pan onto a sieve of the fine-mesh size designated on the label, shake for 3 min on the mechanical sieve shaker, and weigh. Acceptance criteria: The percentage of Barium Hydroxide Lime retained on the screen is NLT 75.0%, and represents the hardness. CARBON DIOXIDE ABSORBENCY Analysis: Fill the lower transverse section of a U-shaped drying tube of about 15-mm internal diameter and 15-cm height with loosely packed glass wool. Place in one arm of the tube, about 5 g of anhydrous calcium chloride, and weigh the tube and the contents. Into the other arm of the tube, place 9.5 g10.5 g of Barium Hydroxide Lime, and again weigh. Insert stoppers in the open arms of the U-tube, and connect the side tube of the arm filled with Barium Hydroxide Lime to a calcium chloride drying tube, which in turn is connected to a suitable source of supply of carbon dioxide. Pass the carbon dioxide through the U-tube at a rate of 75 mL/min for 20 min, timed. Disconnect the Utube, cool to room temperature, remove the stoppers, and weigh.
Barium Sulfate
(Comment on this Monograph)id=m7110=Barium Sulfate=BMonos.pdf) BaSO4 Sulfuric acid, barium salt (1:1); Barium sulfate (1:1) [7727-43-7]. DEFINITION Barium Sulfate contains NLT 97.5% and NMT 100.5% of BaSO4. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL 191, Sulfate Sample solution: Mix 0.5 g of Barium Sulfate with 2 g each of anhydrous sodium carbonate and anhydrous potassium carbonate, heat the mixture in a crucible until fusion is complete, treat the resulting fused mass with hot water, and filter. Acceptance criteria: The filtrate, acidified with hydrochloric acid, meets the requirements. B. IDENTIFICATION TESTSGENERAL 191, Barium Sample solution: Dissolve a portion of the well-washed residue from Identification test A in 6 N acetic acid. Acceptance criteria: The solution meets the requirements. ASSAY PROCEDURE Sample: 0.58 g0.62 g, weighed in a tared platinum crucible Analysis: Add 10 g of anhydrous sodium carbonate to the crucible, and mix by rotating the crucible. Fuse over a blast burner until a clear melt is obtained, and heat for an additional 30 min. Cool, place the crucible in a 400-mL beaker, add 250 mL of water, stir with a glass rod, and heat to dislodge the melt. Remove the crucible from the beaker, and wash with water, collecting the washings in the beaker. Rinse the inside of the crucible with 2 mL of 6 N acetic acid and then with water, again collecting the washings in the beaker, and continue heating and stirring until the melt is disintegrated. Cool the beaker in an ice bath until the precipitate settles, decant the clear liquid through filter paper (Whatman No. 40, or equivalent), taking care to transfer as little precipitate as possible to the paper. Wash twice by decantation as follows. Wash down the inside of the beaker with 10 mL of cold sodium carbonate solution (1 in 50), swirl the contents of the beaker, allow the precipitate to settle, and decant the supernatant through the same filter paper as before, transferring as little precipitate as possible. Place the beaker containing the bulk of the barium carbonate precipitate under the funnel, wash the filter paper with five 1-mL portions of 3 N hydrochloric acid, and wash the paper with water. [NOTEThe solution may be slightly hazy.] Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL of ammonium acetate solution (2 in 5), 25 mL of potassium dichromate solution (1 in 10), and 10.0 g of urea. Cover the beaker with a watch glass, and digest at 80 to 85 for NLT 16 h. Filter while hot through a tared, 233.39
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90.0% and NMT 110.0% of the labeled amount of barium sulfate (BaSO4). It may contain one or more suitable colors, flavors, suspending or dispersing agents, and preservatives. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Sulfate 191 Sample: Ignite a quantity of Paste equivalent to 0.5 g of barium sulfate to constant weight. Analysis: Mix 0.5 g of the ignited Sample with 2 g each of anhydrous sodium carbonate and anhydrous potassium carbonate, heat the mixture in a crucible until fusion is complete, treat the resulting fused mass with hot water, and filter. Proceed as directed. Acceptance criteria: The filtrate, acidified with hydrochloric acid, meets the requirements. B. IDENTIFICATION TESTSGENERAL, Barium 191 Sample solution: Dissolve a portion of the well-washed residue from Identification test A in 6 N acetic acid. Acceptance criteria: The solution meets the requirements. ASSAY PROCEDURE Sample: Barium Sulfate Paste, equivalent to 0.60 g of barium sulfate, weighed in a tared platinum crucible Analysis: Ignite the Sample over a low flame until any organic matter is thoroughly carbonized. Cool, cautiously add 0.5 mL of nitric acid and 0.5 mL of sulfuric acid, and continue the ignition over a low flame until the residue becomes gray in color, then ignite over the full heat of a blast burner. Allow the contents of the crucible to cool to room temperature. [NOTEIf the specimen contains a silicate, such as bentonite, proceed as follows. Add 10 mL of water and 1 mL of sulfuric acid to the residue in the crucible, mix, and add 10 mL of hydrofluoric acid. Heat gently over a low flame until fumes of sulfur trioxide appear. Add 5 mL more of hydrofluoric acid, heat again over a low flame to the appearance of dense fumes, and continue heating until the sulfuric acid has been completely volatilized. Allow the contents of the crucible to cool.] [NOTEIf the specimen does not contain a silicate, omit the treatment of the specimen with hydrofluoric and sulfuric acids.] Add to the treated or untreated specimen in the platinum crucible, 10 g of anhydrous sodium carbonate, fuse over a blast burner until a clear melt is obtained, and heat for an additional 30 min. Cool, place the crucible in a 400-mL beaker, add 250 mL of water, stir with a glass rod, and heat to dislodge the melt. Remove the crucible from the beaker, and wash with water, collecting the washings in the beaker. Rinse the inside of the crucible with 2 mL of 6 N acetic acid and then with water, again collecting the washings in the beaker, and continue heating and stirring until the melt is disintegrated. Cool the beaker in an ice bath until the precipitate settles, decant the clear liquid through filter paper (Whatman No. 40, or equivalent), taking care to transfer as little precipitate as possible to the paper. Wash twice by decantation as follows. Wash down the inside of the beaker with 10 mL of cold sodium carbonate solution (1 in 50), swirl the contents of the beaker, allow the precipitate to settle, and decant the supernatant through the same filter paper as before, transferring as little precipitate as possible. Place the beaker containing the bulk of the barium carbonate precipitate under the funnel, wash the filter paper with five 1-mL portions of 3 N hydrochloric acid, and wash the paper with water. [NOTEThe solution may be slightly hazy.] Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL of ammonium acetate solution (2 in 5), 25 mL of potassium dichromate solution (1 in 10), and 10.0 g of
fine-porosity, sintered-glass crucible, transferring all of the precipitate with the aid of a rubber-tipped stirring rod. Wash the precipitate with potassium dichromate solution (1 in 200), and finally with 20 mL of water. Dry at 105 for 2 h, cool, and weigh. The weight of the barium chromate so obtained, multiplied by 0.9213, represents the weight of BaSO4. Acceptance criteria: 97.5%100.5% of BaSO4 IMPURITIES Inorganic Impurities HEAVY METALS 231: NMT 10 ppm Sample solution: Boil 4.0 g with a mixture of 2 mL of glacial acetic acid and 48 mL of water for 10 min. Dilute with water to 50 mL, filter, and use 25 mL of the filtrate. LIMIT OF SULFIDE Sample solution: Transfer 10 g to a 500-mL conical flask. Add 100 mL of 0.3 N hydrochloric acid. Control: 100 mL of 0.3 N hydrochloric acid containing 5 g of sulfide in a 500-mL conical flask Analysis: Cover the mouth of each conical flask with a circle of filter paper that has been moistened at the area over the mouth of the flask with 0.15 mL of lead acetate TS, the paper being held in place with a string tied around the neck of the flask. Boil each mixture gently for 10 min, taking care to avoid spattering the paper. Acceptance criteria: Any darkening of the paper is not greater than that produced by the similarly treated Control (NMT 0.5 ppm). LIMIT OF ACID-SOLUBLE SUBSTANCES Sample solution: Cool the mixture obtained in the test for Limit of Sulfide, add water to restore approximately the original volume, and filter it through paper that previously has been washed with a mixture of 10 mL of 3 N hydrochloric acid and 90 mL of water, returning the first portions, if necessary, to obtain a clear filtrate. Analysis: Evaporate 50 mL of the filtrate on a steam bath to dryness, and add 2 drops of hydrochloric acid and 10 mL of hot water. Filter again through acid-washed paper, prepared as directed above, wash the filter with 10 mL of hot water, and evaporate the combined filtrate and washings in a tared dish on a steam bath to dryness. Acceptance criteria: The residue, when dried at 105 for 1 h, weighs NMT 15 mg (NMT 0.3%). LIMIT OF SOLUBLE BARIUM SALTS Sample: Residue obtained in the test for Limit of AcidSoluble Substances Control: 10 mL of water containing 0.5 mL of 2 N sulfuric acid and 50 g of barium Analysis: Treat the Sample with 10 mL of water, pass the solution through a filter previously washed with 100 mL of 0.3 N hydrochloric acid, and add 0.5 mL of 2 N sulfuric acid. Acceptance criteria: Any turbidity formed within 30 min is NMT that produced in the similarly treated Control (NMT 0.001%). SPECIFIC TESTS PH 791: 3.510.0, in a 10% (w/w) aqueous suspension ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Barium Sulfate Paste

(Comment on this Monograph)id=m7120=Barium Sulfate Paste=B-Monos.pdf) DEFINITION Barium Sulfate Paste is a semisolid formulation of finely divided particles of Barium Sulfate in a suitable base. It contains NLT
USP 32
urea. Cover the beaker with a watch glass, and digest at 8085 for NLT 16 h. Filter while hot through a tared, fine-porosity, sintered-glass crucible, transferring all of the precipitate with the aid of a rubber-tipped stirring rod. Wash the precipitate with potassium dichromate solution (1 in 200), and finally with 20 mL of water. Dry at 105 for 2 h, cool, and weigh. The weight of the barium chromate so obtained, multiplied by 0.9213, represents the weight of BaSO4. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: For products labeled for oral administration or labeled for oral and rectal administration, the total aerobic microbial count does not exceed 100 cfu/g, and the total combined molds and yeasts count does not exceed 10 cfu/g. For products labeled for rectal administration, the total aerobic microbial count does not exceed 1000 cfu/g, and the total combined molds and yeasts count does not exceed 100 cfu/g. For all products, it meets the requirements of the tests for absence of Salmonella species, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, and the total enterobacterial count does not exceed 10 cfu/g. PH 791: 3.010.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from freezing and from excessive heat.
Official Monographs / Barium 15

acid and 0.5 mL of sulfuric acid, and continue the ignition over a low flame until the residue becomes gray in color, then ignite over the full heat of a blast burner. Allow the contents of the crucible to cool to room temperature. [NOTEIf the specimen contains a silicate, such as bentonite, proceed as follows. Add 10 mL of water and 1 mL of sulfuric acid to the residue in the crucible, mix, and add 10 mL of hydrofluoric acid. Heat gently over a low flame until fumes of sulfur trioxide appear. Add 5 mL more of hydrofluoric acid, heat again over a low flame to the appearance of dense fumes, and continue heating until the sulfuric acid has been completely volatilized. Allow the contents of the crucible to cool.] [NOTEIf the specimen does not contain a silicate, omit the treatment of the specimen with hydrofluoric and sulfuric acids.] Add to the treated or untreated specimen in the platinum crucible, 10 g of anhydrous sodium carbonate, fuse over a blast burner until a clear melt is obtained, and heat for an additional 30 min. Cool, place the crucible in a 400-mL beaker, add 250 mL of water, stir with a glass rod, and heat to dislodge the melt. Remove the crucible from the beaker, and wash with water, collecting the washings in the beaker. Rinse the inside of the crucible with 2 mL of 6 N acetic acid and then with water, again collecting the washings in the beaker, and continue heating and stirring until the melt is disintegrated. Cool the beaker in an ice bath until the precipitate settles, decant the clear liquid through filter paper (Whatman No. 40, or equivalent), taking care to transfer as little precipitate as possible to the paper. Wash twice by decantation as follows. Wash down the inside of the beaker with 10 mL of cold sodium carbonate solution (1 in 50), swirl the contents of the beaker, allow the precipitate to settle, and decant the supernatant through the same filter paper as before, transferring as little precipitate as possible. Place the beaker containing the bulk of the barium carbonate precipitate under the funnel, wash the filter paper with five 1-mL portions of 3 N hydrochloric acid, and wash the paper with water. [NOTEThe solution may be slightly hazy.] Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL of ammonium acetate solution (2 in 5), 25 mL of potassium dichromate solution (1 in 10), and 10.0 g of urea. Cover the beaker with a watch glass, and digest at 80 to 85 for NLT 16 h. Filter while hot through a tared, fine-porosity, sintered-glass crucible, transferring all of the precipitate with the aid of a rubber-tipped stirring rod. Wash the precipitate with potassium dichromate solution (1 in 200), and finally with 20 mL of water. Dry at 105 for 2 h, cool, and weigh. The weight of the barium chromate so obtained, multiplied by 0.9213, represents the weight of BaSO4. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: The total bacterial count does not exceed 100 cfu/mL; the total combined molds and yeasts count does not exceed 10 cfu/mL; and it meets the requirements of the tests for absence of Salmonella species, Staphylococcus aureus, and Pseudomonas aeruginosa. PH 791: 3.510.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing.
Barium Sulfate Suspension

(Comment on this Monograph)id=m7135=Barium Sulfate Suspension=B-Monos.pdf) DEFINITION Barium Sulfate Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of barium sulfate (BaSO4). It contains suitable dispersing and/or suspending agents so that when mixed as directed in the labeling, it yields a uniformly dispersed suspension. It may contain one or more suitable colors, flavors, fluidizing agents, and preservatives. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL 191, Sulfate Sample: Shake the Suspension and transfer a volume equivalent to 0.5 g of barium sulfate to a suitable container. Ignite to constant weight. Analysis: Mix 0.5 g of the ignited Sample with 2 g each of anhydrous sodium carbonate and anhydrous potassium carbonate, heat the mixture in a crucible until fusion is complete, treat the resulting fused mass with hot water, and filter. Proceed as directed. Acceptance criteria: The filtrate, acidified with hydrochloric acid, meets the requirements. B. IDENTIFICATION TESTSGENERAL 191, Barium Sample solution: Dissolve a portion of the well-washed residue from Identification test A in 6 N acetic acid. Acceptance criteria: The solution meets the requirements. ASSAY PROCEDURE Sample: A volume of Suspension, previously well shaken in its original container, equivalent to 0.60 g of barium sulfate, in a tared platinum crucible Analysis: Ignite over a low flame until any organic matter is thoroughly carbonized. Cool, cautiously add 0.5 mL of nitric
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Barium / Official Monographs
USP 32
portions of 3 N hydrochloric acid, and wash the paper with water. [NOTEThe solution may be slightly hazy.] Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL of ammonium acetate solution (2 in 5), 25 mL of potassium dichromate solution (1 in 10), and 10.0 g of urea. Cover the beaker with a watch glass, and digest at 80 to 85 for NLT 16 h. Filter while hot through a tared, fine-porosity, sintered-glass crucible, transferring all of the precipitate with the aid of a rubber-tipped stirring rod. Wash the precipitate with potassium dichromate solution (1 in 200), and finally with 20 mL of water. Dry at 105 for 2 h, cool, and weigh. The weight of the barium chromate so obtained, multiplied by 0.9213, represents the weight of BaSO4. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS LOSS ON DRYING 731: Dry at 105 for 4 h: it loses NMT 1.0% of its weight. PH 791: 3.510.0, in a 60% (w/w) aqueous suspension, or constituted for its intended use as directed in the labeling ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Barium Sulfate for Suspension

(Comment on this Monograph)id=m7140=Barium Sulfate for Suspension=B-Monos.pdf) DEFINITION Barium Sulfate for Suspension is a dry mixture of Barium Sulfate and one or more suitable dispersing and/or suspending agents. It contains NLT 90.0% and NMT 110.0% of the labeled amount of barium sulfate (BaSO4). It may contain one or more suitable colors, flavors, fluidizing agents, and preservatives. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL 191, Sulfate Sample: Ignite 1g to constant weight Analysis: Mix 0.5 g of the ignited Sample with 2 g each of anhydrous sodium carbonate and anhydrous potassium carbonate, heat the mixture in a crucible until fusion is complete, treat the resulting fused mass with hot water, and filter. Acceptance criteria: The filtrate, acidified with hydrochloric acid, meets the requirements. B. IDENTIFICATION TESTSGENERAL191, Barium Sample solution: Dissolve a portion of the well-washed residue from Identification test A in 6 N acetic acid. Acceptance criteria: The solution meets the requirements. ASSAY PROCEDURE Sample: Barium Sulfate for Suspension, equivalent to 0.60 g of barium sulfate, weighed in a tared platinum crucible Analysis: Ignite over a low flame until any organic matter is thoroughly carbonized. Cool, cautiously add 0.5 mL of nitric acid and 0.5 mL of sulfuric acid, and continue the ignition over a low flame until the residue becomes gray in color, then ignite over the full heat of a blast burner. Allow the contents of the crucible to cool to room temperature. [NOTEIf the specimen contains a silicate, such as bentonite, proceed as follows. Add 10 mL of water and 1 mL of sulfuric acid to the residue in the crucible, mix, and add 10 mL of hydrofluoric acid. Heat gently over a low flame until fumes of sulfur trioxide appear. Add 5 mL more of hydrofluoric acid, heat again over a low flame to the appearance of dense fumes, and continue heating until the sulfuric acid has been completely volatilized. Allow the contents of the crucible to cool.] [NOTEIf the specimen does not contain a silicate, omit the treatment of the specimen with hydrofluoric and sulfuric acids.] Add to the treated or untreated specimen in the platinum crucible, 10 g of anhydrous sodium carbonate, fuse over a blast burner until a clear melt is obtained, and heat for an additional 30 min. Cool, place the crucible in a 400-mL beaker, add 250 mL of water, stir with a glass rod, and heat to dislodge the melt. Remove the crucible from the beaker, and wash with water, collecting the washings in the beaker. Rinse the inside of the crucible with 2 mL of 6 N acetic acid and then with water, again collecting the washings in the beaker, and continue heating and stirring until the melt is disintegrated. Cool the beaker in an ice bath until the precipitate settles, decant the clear liquid through filter paper (Whatman No. 40, or equivalent), taking care to transfer as little precipitate as possible to the paper. Wash twice by decantation as follows. Wash down the inside of the beaker with 10 mL of cold sodium carbonate solution (1 in 50), swirl the contents of the beaker, allow the precipitate to settle, and decant the supernatant through the same filter paper as before, transferring as little precipitate as possible. Place the beaker containing the bulk of the barium carbonate precipitate under the funnel, wash the filter paper with five 1-mL
Barium Sulfate Tablets

(Comment on this Monograph)id=m7155=Barium Sulfate Tablets=B-Monos.pdf) DEFINITION Barium Sulfate Tablets are flat-sided disks between 11.5 mm and 13.5 mm in diameter and contain NLT 90.0% and NMT 110.0% of the labeled amount of BaSO4. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Sulfate 191 Sample: A portion of powdered Tablets equivalent to 0.6 g of barium sulfate Analysis: Mix the Sample with 2 g each of anhydrous sodium carbonate and anhydrous potassium carbonate, heat the mixture in a crucible until fusion is complete, treat the resulting fused mass with hot water, and filter. Proceed as directed. Acceptance criteria: The filtrate, acidified with hydrochloric acid, meets the requirements. B. IDENTIFICATION TESTSGENERAL, Barium 191 Sample solution: Dissolve a portion of the well-washed residue from Identification test A in 6 N acetic acid. Acceptance criteria: The solution meets the requirements. ASSAY PROCEDURE Sample: A portion of powdered Tablets, equivalent to 0.6 g of barium sulfate, weighed in a tared platinum crucible Analysis: Add 10 g of anhydrous sodium carbonate to the crucible, and mix by rotating the crucible. Fuse over a blast burner until a clear melt is obtained, and heat for an additional 30 min. Cool, place the crucible in a 400-mL beaker, add 250 mL of water, stir with a glass rod, and heat to dislodge the melt. Remove the crucible from the beaker, and wash with water, collecting the washings in the beaker. Rinse the inside of the crucible with 2 mL of 6 N acetic acid and then with water, again collecting the washings in the beaker, and continue heating and stirring until the melt is disintegrated. Cool the beaker in an ice bath until the precipitate settles, decant the clear liquid through filter paper (Whatman No. 40, or equivalent), taking care to transfer as little precipitate as possible to the paper. Wash
USP 32
twice by decantation as follows. Wash down the inside of the beaker with 10 mL of cold sodium carbonate solution (1 in 50), swirl the contents of the beaker, allow the precipitate to settle, and decant the supernatant through the same filter paper as before, transferring as little precipitate as possible. Place the beaker containing the bulk of the barium carbonate precipitate under the funnel, wash the filter paper with five 1-mL portions of 3 N hydrochloric acid, and wash the paper with water. [NOTEThe solution may be slightly hazy.] Add 100 mL of water, 5.0 mL of hydrochloric acid, 10.0 mL of ammonium acetate solution (2 in 5), 25 mL of potassium dichromate solution (1 in 10), and 10.0 g of urea. Cover the beaker with a watch glass, and digest at 8085 for NLT 16 h. Filter while hot through a tared, fine-porosity, sintered-glass crucible, transferring all of the precipitate with the aid of a rubber-tipped stirring rod. Wash the precipitate with potassium dichromate solution (1 in 200), and finally with 20 mL of water. Dry at 105 for 2 h, cool, and weigh. The weight of the barium chromate so obtained, multiplied by 0.9213, represents the weight of BaSO4. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701 Time: NLT 10 min and NMT 30 min UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Official Monographs / BCG 17

total of at least 4 mg of the Sample solution intramuscularly or subcutaneously in the rear left internal thigh, and observe them for a period of 6 weeks. Note the number of animals that survive at the end of the observation period, and then sacrifice them. Perform autopsies of all animals postmortem to examine them for evidence of tuberculous infections, particularly at the popliteal and inguinal lymph nodes, liver, spleen, pancreas, and lungs, as well as at the injection site. If any abnormalities are found, perform a histological examination using standard and Acid-Fast staining techniques to detect Acid-Fast organisms. Acceptance criteria: The product complies with the test if none of the animals show signs of tuberculosis and NMT one-third of the animals die during the observation period. SKIN REACTIVITY Sample solutions: Using the same diluent and the Sample solution, prepared as directed in the test for Virulent Mycobacteria, further dilute aseptically by making three serial tenfold dilutions. Analysis: Randomly select two guinea pigs (male or female), each weighing 250300 g. Inject 0.1 mL of each of the four suspensions intradermally at different sites on the back of each animal. After 4 weeks, the animals are shaved so that the injection sites and any reactions are made clearly visible. The diameters of the reactions are measured, and the presence of necrosis or nodules are noted. Acceptance criteria: The reaction for the largest dose is between 410 mm and the smallest dose induces a nodule less than or equal to 4 mm. Each animal gains weight during the observation period. TUBERCULIN SENSITIVITY Tuberculin solution: Use tuberculin, purified protein derivative, to prepare a solution containing 25 U.S. Tuberculin Units/0.1 mL. Dilute aseptically, if necessary, with sterile 0.9% sodium chloride solution. Analysis: Use the same animals on which the Skin Reactivity test is performed. After the Skin Reactivity test is completed, inject each animal intradermally on the back with 0.1 mL of the Tuberculin solution, and observe after 1824 h. Acceptance criteria: An erythematous reaction of NLT 10 mm in diameter is measured on each animal. RESIDUAL MOISTURE: NMT the limit approved for the particular product, determined by a suitable validated method Limits vary in accordance with the method. VIABILITY: Determine the potencies of BCG Live using NLT 5 containers before freeze-drying, and an equal number of containers after freeze-drying, following the procedure under Potency, except use the suspension before freeze-drying as is. The loss in viability due to freeze-drying is NMT 90%. POTENCY: Determine the number of viable units/mL by viable count on solid medium using a method suitable for the product to be examined. Alternatively, a validated biochemical method may be used.
BCG Live
(Comment on this Monograph)id=m703=BCG Live=BMonos.pdf) DEFINITION BCG Live (intravesical) for immunotherapy is a freeze-dried solution of attenuated live bacteria derived from a culture of Bacillus Calmette-Gu rin (Mycobacterium bovis, var. BCG) and e used intravesically in the treatment of carcinoma in situ and papilloma tumors of the urinary bladder. The bacteria are grown in a medium that does not contain substances known to cause toxic or allergic reactions in human beings or to cause the bacteria to become virulent for guinea pigs. The culture is harvested and formulated to contain one or more excipients. The freeze-dried solution is reconstituted and further diluted aseptically with a sterile diluent for use. A reconstituted dose contains 1.0-19.2 108 colony-forming units (cfu). BCG Live does not contain a preservative. IDENTIFICATION BCG Live is identified by microscopic examination of the bacilli in stained smears demonstrating their acid-fast property. Alternatively, validated molecular biology techniques may be used. SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Direct Inoculation of the Culture Medium. GENERAL SAFETY: It meets the requirements as set forth for under Biological Reactivity Tests, In Vivo 88, Safety Tests Biologicals, modified as follows. Guinea pigs are injected intraperitoneally with 3.0 mL of the reconstituted product. VIRULENT MYCOBACTERIA Sample solution: Reconstitute the freeze-dried BCG Live as per the manufacturers instructions for human use with the diluent recommended by the manufacturer, and dilute aseptically to about 2 mg/mL with sterile BCG diluents. Analysis: Randomly select NLT six guinea pigs of the same sex, each weighing 250300 g. Inject each animal with a
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: BCG Live is sensitive to light and therefore must be preserved and stored in a glass container where it is protected from direct light at a temperature between 2 and 8. EXPIRATION DATE: The product is stable for 3 years when stored between 2 and 8. LABELING: Label it to indicate the dry weight of bacteria in a vial, cfu/dose, the storage conditions, the expiration date, and that it is not to be used after the expiration date given on the package. Label it to state that it should be protected from light and that it should be used immediately after reconstitution/dilution. Label it to indicate that it is for intravesical use.
18
BCG / Official Monographs
USP 32
Sample solution: Transfer 4.0 mL of Sample stock solution to a suitable vial, and add 4.0 mL of Internal standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 and a pump capable of operating at a column pressure of up to 3500 psi [NOTEAdjust the specimen size and other operating parameters such that the peak from the internal standard in the Standard solution is about 0.60.9 full-scale.] Injection size: 525 L System suitability Sample: Standard solution [NOTEThe retention time of beclomethasone dipropionate is about 6 min, and that of testosterone propionate is about 10 min.] Suitability requirements Relative standard deviation: NMT 3.0% for five replicate injections Analysis: Samples: Sample solution and Standard solution Calculate the percentage of C28H37ClO7 in the portion of Beclomethasone Dipropionate taken: Result = (RU/RS) (CS/CU) 100 = peak height ratio of beclomethasone dipropionate to the internal standard from the Sample solution = peak height ratio of beclomethasone RS dipropionate to the internal standard from the Standard solution = concentration of USP Beclomethasone CS Dipropionate RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 97.0%103.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: RU
BCG Vaccine
(Comment on this Monograph)id=m7200=BCG Vaccine=BMonos.pdf) DEFINITION BCG Vaccine conforms to the regulations of the FDA concerning biologics (see Biologics 1041). It is a dried, living culture of the bacillus Calmette-Gu rin strain of Mycobacterium e tuberculosis var. bovis, grown in a suitable medium from a seed strain of known history that has been maintained to preserve its capacity for conferring immunity. It contains an amount of viable bacteria such that inoculation, in the recommended dose, of tuberculin-negative persons results in an acceptable tuberculin conversion rate. It is free from other organisms, and contains a suitable stabilizer. It contains no antimicrobial agent. [NOTEUse the Vaccine immediately after its constitution, and discard any unused portion after 2 h.] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in hermetic containers, preferably of Type I glass, at a temperature between 2 and 8. EXPIRATION DATE: The expiration date is not later than 6 months after date of issue, or not later than 1 year after date of issue if stored at a temperature below 5.
Beclomethasone Dipropionate
(Comment on this Monograph)id=m7250=Beclomethasone Dipropionate=B-Monos.pdf)
C28H37ClO7 Pregna-1,4-diene-3,20-dione, 9-chloro-11-hydroxy-16methyl-17,21-bis(1-oxopropoxy)-, (11,16)-; 9-Chloro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 17,21-dipropionate [5534-09-8]. Monohydrate
521.04
NMT 0.1%
539.07
DEFINITION Beclomethasone Dipropionate is anhydrous or contains one molecule of water of hydration. It contains NLT 97.0% and NMT 103.0% of C28H37ClO7, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197M ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:2) Internal standard solution: 1.2 mg/mL of USP Testosterone Propionate RS in methanol Standard stock solution: 1.4 mg/mL of USP Beclomethasone Dipropionate RS in methanol Standard solution: Transfer 4.0 mL of Standard stock solution to a suitable vial, and add 4.0 mL of Internal standard solution to obtain a solution having a known concentration of about 0.7 mg/mL of USP Beclomethasone Dipropionate RS and 0.6 mg/mL of USP Testosterone Dipropionate RS. Sample stock solution: 1.4 mg/mL of Beclomethasone Dipropionate in methanol
SPECIFIC TESTS SPECIFIC ROTATION 781S: +88 to +94 Sample solution: 10 mg/mL, in dioxane LOSS ON DRYING 731: Dry it at 105 for 3 h: the anhydrous form loses NMT 0.5% of its weight; the monohydrate form loses between 2.8% and 3.8% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS USP Testosterone Propionate RS
Belladonna Leaf
(Comment on this Monograph)id=m7410=Belladonna Leaf=BMonos.pdf) DEFINITION Belladonna Leaf consists of the dried leaf and flowering or fruiting top of Atropa belladonna Linn or of its variety e acuminata Royle ex Lindley (Fam. Solanaceae). Belladonna Leaf yields NLT 0.35% of the alkaloids of belladonna leaf.
USP 32
ASSAY PROCEDURE Phosphate buffer: 34.8 g of dibasic potassium phosphate in 900 mL of water Adjust to a pH of 9.5 by the addition of 3 N hydrochloric acid or sodium hydroxide, with mixing. Diluent: Dilute sulfuric acid (1 in 350) Internal standard solution: 0.8 mg/mL of USP Homatropine Hydrobromide RS in Diluent [NOTEPrepare fresh on the day of use.] Standard stock solution A: 1.0 mg/mL of USP Scopolamine Hydrobromide RS in Diluent Standard stock solution B: Dissolve 20 mg of USP Atropine Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask, add 2.0 mL of Standard stock solution A, and mix. Add Diluent to volume. [NOTEPrepare fresh on the day of use.] Standard solutions: Pipet into three separate 60-mL separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Standard stock solution A, and add 9.0, 8.0, and 7.0 mL, respectively, of Diluent. Add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake vigorously, allow the layers to separate, and discard the chloroform layer. [NOTEIf emulsions are formed, a mixed solvent consisting of chloroform-isopropyl alcohol (10:3) may be substituted for chloroform throughout the procedure.] Add another 15 mL of chloroform, and extract again, discarding the chloroform phase. Add 15 mL of Phosphate buffer and sufficient 1 N sodium hydroxide to yield a final pH between 9.0 and 9.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1 mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Sample solution: Moisten 10 g, previously reduced to a moderately coarse powder, with a mixture of 8 mL of ammonium hydroxide, 10 mL of alcohol, and 20 mL of ether, and extract the alkaloids by either Method I or Method II below. If necessary, reduce the volume of the extract to 100 mL by evaporation on a steam bath. Method I: Place the moistened drug in a continuousextraction thimble, and allow maceration to proceed overnight, then extract with ether for 3 h, or longer if necessary to effect complete extraction. Method II: Place the moistened drug in a small percolator, and allow maceration to proceed overnight. Percolate slowly with a mixture of 3 volumes of ether and 1 volume of chloroform. Continue the percolation until the residue from 3 to 4 mL of percolate last passed, when dissolved in dilute sulfuric acid (1 in 70) and treated with mercuric iodide TS, shows NMT a faint turbidity. Transfer the extract from Method I or Method II to a separator with the aid of ether. Extract with five 15-mL portions of dilute sulfuric acid (1 in 70), filtering each portion drawn off into a 100-mL volumetric flask. Wash the filter with dilute sulfuric acid (1 in 70), and collect the washings in the flask. Add dilute sulfuric acid (1 in 70) to volume, and mix. Dilute 20.0 mL of the resulting solution with the same dilute acid to 100.0 mL. Pipet 10 mL of this solution into a 60-mL separator. Add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake
Official Monographs / Belladonna 19

vigorously, allow the layers to separate, and discard the chloroform layer. [NOTEIf emulsions are formed, a mixed solvent consisting of chloroform-isopropyl alcohol (10:3) may be substituted for chloroform throughout the extraction procedure.] Add another 15 mL of chloroform, and extract again, discarding the chloroform phase. Add 15 mL of Phosphate buffer and sufficient 1 N sodium hydroxide to yield a final pH between 9.0 and 9.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate (see Suitability for alkaloid assays under Reagents, Indicators, and SolutionsSodium Sulfate, Anhydrous), previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1 mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Extraction blank: Place 10 mL of Diluent in a 60-mL separator. Proceed as directed under Sample solution, beginning with then add 15 mL of chloroform. The blank chromatogram contains no significant interferences at the locus of atropine, scopolamine, or homatropine. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.2-m 4-mm; glass column packed with 3% G3 on S1AB [NOTEThe column may be cured and conditioned as specified under Chromatography 621, Gas Chromatography.] Temperature Column: 215 Injector port: 240 Detector: 240 Carrier gas: Dry helium Flow rate: 65 mL/min Injection size: 5 L System suitability Sample: Sample solution (610 injections) Suitability requirements Resolution: NLT 3.0 between aH and aA (R) Tailing factor: NMT 2.0 (the sum of the distances from peak center to the leading edge and to the tailing edge divided by twice the distance from peak center to the leading edge), measured at 5% of the peak height of aA Relative standard deviation: The analytical system is suitable for conducting this Assay if the relative standard deviation for the ratio, RA, is NMT 2.0%, calculated as follows: (standard deviation/mean ratio) 100 Analysis Samples: Standard solutions and Sample solution Measure the areas, aA, aH, and aS, of the atropine, homatropine, and scopolamine peaks, respectively, in each chromatogram, and calculate the ratios AA and AS: Result = aA/aH and aS/aH Plot the standard curves of the values of RA and RS against the amounts, in mg, of atropine and scopolamine in the solutions. (The ratio of the molecular weight of atropine
20
Belladonna / Official Monographs
USP 32
having three germinal furrows and rows of pits between the ridges on the exine. Fruit: The epicarp exhibits polygonal epidermal cells with a striated cuticle and stomata. The mesocarp consists of large pulp cells some of which contain rosette aggregate crystals of calcium oxalate. Seed: The seed is characterized by an epidermis of large, wavy-walled cells with prominent ridges over the anticlinal walls. Powdered belladonna leaf: Light olive-brown to moderate olive-green in color. The following are among the elements of identification: the separate microcrystals, the dark gray crystal cells, the cuticular striping of the epidermal cells, the vessels with ellipsoidal bordered pits, the fibers of the stem, and occasional hairs and pollen grains. Rosette aggregates of calcium oxalate and fragments of the seed occur when the drug contains belladonna fruits. Examine Belladonna Leaf for hairs having a papillose cuticle and for raphides of calcium oxalate: their presence indicates adulteration. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers and avoid long exposure to direct sunlight. Preserve powdered Belladonna Leaf in light-resistant containers. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS USP Homatropine Hydrobromide RS USP Scopolamine Hydrobromide RS
to that of anhydrous atropine sulfate is 0.8551, and the ratio of the molecular weight of scopolamine to that of anhydrous scopolamine hydrobromide is 0.7894.) Inject a portion of the Sample solution into the chromatograph, obtain the chromatogram area ratios, measure the peak areas, and calculate the area ratios, as with the Standard solutions. Record from the standard curves the quantities, in mg, of atropine and scopolamine in the weight of the specimen taken. Add the quantity, in mg, of atropine and scopolamine, and multiply by 50 to obtain the weight, in mg, of alkaloids in the portion of Belladonna Leaf taken. Acceptance criteria: NLT 0.35% of the alkaloids of belladonna leaf IMPURITIES ARTICLES OF BOTANICAL ORIGIN, Acid-Insoluble Ash 561: NMT 3.0% BELLADONNA STEMS: The proportion of belladonna stems over 10 mm in diameter does not exceed 3.0%. SPECIFIC TESTS BOTANIC CHARACTERISTICS Belladonna leaf: Usually partly matted together, crumpled or broken leaves, together with some smaller stems and a number of flowers and fruits The leaves are thin and brittle, mostly light green to moderate olive-green. The lamina is mostly from 5 to 25 cm in length and from 4 to 12 cm in width and possesses an ovate-lanceolate to broadly ovate outline, an acute to acuminate apex, an entire margin, an acute to somewhat decurrent base and slightly hairy surface, the hairs being more abundant along the veins; when broken transversely, it shows numerous light-colored dots (crystal cells) visible with a lens. The petiole is slender and usually up to 4 cm in length. The flowers possess a campanulate corolla with five small, reflexed lobes, purplish to yellowish purple, becoming faded to brown or dusky yellow or yellow; a green, five-lobed calyx; five epipetalous stamens; and a superior, bilocular ovary with numerous ovules. The fruit is subglobular, dark yellow to yellowish brown to dusky red or black, up to 12 mm in width, and sometimes subtended by the persistent calyx and containing numerous flattened, somewhat reniform seeds, the latter up to 2 mm in width. The stems are more or less flattened and hollow and finely hairy when young. Histology Leaf: The epidermis of the lamina possesses wavy anticlinal walls and a distinctly striated cuticle. Stomata are more numerous in the lower epidermis and are surrounded by three or four neighboring cells, one of which is smaller than the others. The nonglandular hairs are uniseriate and up to six-celled. Short club-shaped glandular hairs with a one-celled stalk and multicellular head and long glandular hairs with a uniseriate stalk and unicellular head occur on both epidermises. The mesophyll consists of a single layer of palisade parenchyma beneath which occurs spongy parenchyma, the latter with scattered cells filled with microcrystals. The midrib contains an arc of bicollateral bundles, collenchyma beneath upper epidermis, and scattered parenchyma cells with microcrystals. Stem: The stem shows an epidermis with striated cuticle and few hairs; a distinct endodermis; small strands of long, thin-walled, slightly lignified pericyclic fibers; and a circle of bicollateral bundles. The parenchyma of the cortex and pith is interspersed with crystal cells. Flower: The calyx possesses numerous glandular hairs with uniseriate stalks and one- to three-celled glandular heads. The corolla shows a papillose inner epidermis and an outer epidermis with glandular hairs similar to those of the calyx. The pollen grains, when mounted in chloral hydrate solution, are subspherical, 40 m in diameter, tricolpate,
Belladonna Extract
(Comment on this Monograph)id=m7330=Belladonna Extract=B-Monos.pdf) DEFINITION Belladonna Extract contains, in each 100 g, NLT 1.15 g and NMT 1.35 g of the alkaloids of belladonna leaf. Pilular Belladonna Extract Prepare the extract by percolating 1000 g of Belladonna Leaf, using a mixture of 3 volumes of alcohol and 1 volume of water as the menstruum. Macerate the drug for 16 h, and then percolate it at a moderate rate. Evaporate the percolate under reduced pressure and at a temperature not exceeding 60 to a pilular consistency, and adjust the remaining extract, after assaying, by dilution with liquid glucose so that the finished Extract will contain, in each 100 g, 1.25 g of the alkaloids of belladonna leaf. Powdered Belladonna Extract Prepare the extract by percolating 1000 g of Belladonna Leaf, using alcohol as the menstruum. Macerate the drug for 16 h, and then percolate it slowly. Evaporate the percolate under reduced pressure and at a temperature not exceeding 60 to a soft extract, add 50 g of dry starch, and continue the evaporation, at the same temperature, until the product is dry. Powder the residue. The extract may be deprived of its fat by treating either the soft extract first obtained, or the dry and powdered extract, as directed under Pharmaceutical Dosage Forms 1151, Extracts. Assay the powdered residue, and add sufficient starch, previously dried at 100, to obtain a finished Extract containing 1.25 g of the alkaloids of belladonna leaf in each 100 g. Mix the powders, and pass the Extract through a fine sieve. ASSAY PROCEDURE Phosphate buffer: 34.8 g of dibasic potassium phosphate in 900 mL of water Adjust to a pH of 9.5 by the addition of 3 N hydrochloric acid or sodium hydroxide, with mixing.
USP 32
Diluent: Dilute sulfuric acid (1 in 350) Internal standard solution: 0.8 mg/mL of USP Homatropine Hydrobromide RS in Diluent [NOTEPrepare fresh on the day of use.] Standard stock solution A: 1.0 mg/mL of USP Scopolamine Hydrobromide RS in Diluent Standard stock solution B: Dissolve 20 mg of USP Atropine Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask, and add 2.0 mL of Standard stock solution A. Add Diluent to volume. [NOTEPrepare fresh on the day of use.] Standard solutions: Pipet into three separate 60-mL separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Standard solution A, and add 9.0, 8.0, and 7.0 mL, respectively, of Diluent. Add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake vigorously, allow the layers to separate, and discard the chloroform layer. (If emulsions are formed, a mixed solvent consisting of chloroform-isopropyl alcohol (10:3) may be substituted for chloroform throughout the procedure). Add another 15 mL of chloroform, and extract again, discarding the chloroform phase. Add 15 mL of Phosphate buffer and sufficient 1 N sodium hydroxide to yield a final pH between 9.0 and 9.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1 mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Extraction blank: Place 10 mL of Diluent in a 60-mL separator. Proceed as directed under Sample solution, beginning with then add 15 mL of chloroform. The blank chromatogram contains no significant interferences at the locus of atropine, scopolamine, or homatropine. Sample solution: Transfer 0.5 g of Extract to a 125-mL conical flask, and add 40 mL of Diluent. Heat to a temperature not above 45, and stir to hasten solution. Filter the solution through filter paper into a 100-mL volumetric flask. Wash the flask and the filter with two 20-mL portions of warmed Diluent, and collect the washings in the 100-mL volumetric flask. Add Diluent to volume. Pipet 10 mL of this solution into a 60-mL separator. To the separator, add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake vigorously, allow the layers to separate, and discard the chloroform layer. (If emulsions are formed, a mixed solvent consisting of chloroform and isopropyl alcohol (10:3) may be substituted for chloroform throughout the extraction procedure.) Add another 15 mL of chloroform, and extract again, discarding the chloroform phase. Add 15 mL of Phosphate buffer and sufficient 1 N sodium hydroxide to yield a final pH between 9.0 and 9.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate (see Suitability for alkaloid assays under Reagents, Indicators, and SolutionsSodium Sulfate, Anhydrous), previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel

with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1 mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.2-m 4-mm; glass column packed with 3% G3 on S1AB [NOTEThe column may be cured and conditioned as specified under Chromatography 621, Gas Chromatography.] Temperature Column: 215 Injector port: 240 Detector: 240 Carrier gas: Dry helium Flow rate: 65 mL/min Injection size: 5 L System suitability Sample: Standard solutions, six to ten injections Suitability requirements Resolution: NLT 3.0 between aH and aA (R) Tailing factor: NMT 2.0 (the sum of the distances from peak center to the leading edge and to the tailing edge divided by twice the distance from peak center to the leading edge), measured at 5% of the peak height of aA Relative standard deviation: The analytical system is suitable for conducting this assay if the relative standard deviation for the ratio, RA, is NMT 2.0% calculated as follows: (standard deviation/mean ratio) 100 Analysis Samples: Standard solutions and Sample solution Measure the areas, aA, aH, and aS, of the atropine, homatropine, and scopolamine peaks, respectively, in each chromatogram, and calculate the ratios AA and AS: Result = aA/aH and aS/aH Plot the curves of the Standard solutions of the values of RA and RS versus the amounts, in mg, of atropine and scopolamine in the solutions. (The ratio of the molecular weight of atropine to that of anhydrous atropine sulfate is 0.8551, and the ratio of the molecular weight of scopolamine to that of anhydrous scopolamine hydrobromide is 0.7894.) Inject a portion of the Sample solution into the chromatograph, obtain the chromatogram area ratios, measure the peak areas, and calculate the area ratios, as with the Standard solutions. Record from the curves of the Standard solutions the quantities, in mg, of atropine and scopolamine in the volume of specimen taken. Add the quantity, in mg, of atropine and scopolamine, and multiply by 10 to obtain the weight, in mg, of alkaloids in the portion of Extract taken. Acceptance criteria: 1.15 g1.35 g of the alkaloids of belladonna leaf/100 g ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at a temperature not exceeding 30. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS USP Homatropine Hydrobromide RS USP Scopolamine Hydrobromide RS
22
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mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Extraction blank: Place 10 mL of Diluent in a 60-mL separator. Proceed as directed under Sample solution, beginning with then add 15 mL of chloroform. The blank chromatogram contains no significant interferences at the locus of atropine, scopolamine, or homatropine. Sample solution: Transfer an equivalent to 600 g of atropine and scopolamine, from weighed and finely powdered Tablets (NLT 20), to a 60-mL separator, add 10.0 mL of Diluent, and sonicate to dissolve as much as possible of the specimen Add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake vigorously, allow the layers to separate, and discard the chloroform layer. [NOTEIf emulsions are formed, a mixed solvent consisting of chloroform and isopropyl alcohol (10:3) may be substituted for chloroform throughout the extraction procedure.] Add another 15 mL of chloroform, and extract again, discarding the chloroform phase. Add 15 mL of Phosphate Buffer and sufficient 1 N sodium hydroxide to yield a final pH between 9.0 and 9.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate (see Suitability for alkaloid assays under Reagents, Indicators, and SolutionsSodium Sulfate, Anhydrous), previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1 mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.2-m 4-mm; glass column packed with 3% G3 on S1AB [NOTEThe column may be cured and conditioned as specified under Chromatography 621, Gas Chromatography.] Temperature Column: 215 Injector port: 240 Detector: 240 Carrier gas: Dry helium Flow rate: 65 mL/min Injection size: 5 L System suitability Sample: Standard solutions for six to ten injections Suitability requirements Resolution: NLT 3.0 between aH and aA (R) Tailing factor: NMT 2.0 (the sum of the distances from peak center to the leading edge and to the tailing edge divided by twice the distance from peak center to the leading edge), measured at 5% of the peak height of aA Relative standard deviation: The analytical system is suitable for conducting this Assay if the relative standard deviation for the ratio, RA, is NMT 2.0%, calculated by the formula: RA = (standard deviation/mean ratio) 100 Analysis Samples: Standard solutions and Sample solution Measure the areas, aA, aH, and aS, of the atropine, homatropine, and scopolamine peaks, respectively, in each chromatogram, and calculate the ratios AA and AS: Result = aA/aH and aS/aH
Belladonna Extract Tablets

(Comment on this Monograph)id=m7360=Belladonna Extract Tablets=B-Monos.pdf) DEFINITION Belladonna Extract Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of the alkaloids of belladonna leaf. IDENTIFICATION [NOTEThe Sample to be used in Identification tests A and B is prepared as follows.] Sample: Macerate a quantity of powdered Tablets, equivalent to 5 mg of the alkaloids of belladonna extract, with 20 mL of water, and transfer to a separator. Render the solution alkaline with 6 N ammonium hydroxide, and extract the alkaloids with 50 mL of chloroform. Filter the chloroform layer, divide it into two equal portions, and evaporate to dryness. A. PROCEDURE Analysis: To one portion of the Sample add 2 drops of nitric acid, evaporate on a steam bath to dryness, and add a few drops of alcoholic potassium hydroxide TS. Acceptance criteria: A violet color is produced. B. PROCEDURE Analysis: Dissolve the other portion of the Sample in 1 mL of dilute hydrochloric acid (1 in 120), and add gold chloride TS, dropwise with shaking, until a definite precipitate separates. Slowly heat until the precipitate dissolves, and allow the solution to cool. Acceptance criteria: A lusterless precipitate is produced. ASSAY PROCEDURE Phosphate buffer: 34.8 g of dibasic potassium phosphate in 900 mL of water Adjust to a pH of 9.5 by the addition of 3 N hydrochloric acid or sodium hydroxide, with mixing Diluent: Dilute sulfuric acid (1 in 350) Internal standard solution: 0.8 mg/mL of USP Homatropine Hydrobromide RS in Diluent [NOTEPrepare fresh on the day of use.] Standard stock solution A: 1.0 mg/mL of USP Scopolamine Hydrobromide RS in Diluent Standard stock solution B: Dissolve 20 mg of USP Atropine Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask, and add 2.0 mL of Standard stock solution A. Add Diluent to volume. [NOTEPrepare fresh on the day of use.] Standard solutions: Pipet into three separate 60-mL separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Standard stock solution A, and add 9.0, 8.0, and 7.0 mL, respectively, of Diluent. Add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake vigorously, allow the layers to separate, and discard the chloroform layer. [NOTEIf emulsions are formed, a mixed solvent consisting of chloroform-isopropyl alcohol (10:3) may be substituted for chloroform throughout the procedure.] Add another 15 mL of chloroform, and extract again, discarding the chloroform phase. Add 15 mL of Phosphate buffer and sufficient 1 N sodium hydroxide to yield a final pH between 9.0 and 9.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1
USP 32
Plot the curves of the Standard solutions of the values of RA and RS versus the amounts, in mg, of atropine and scopolamine in the solutions. (The ratio of the molecular weight of atropine to that of anhydrous atropine sulfate is 0.8551, and the ratio of the molecular weight of scopolamine to that of anhydrous scopolamine hydrobromide is 0.7894.) Inject a portion of the Sample solution into the chromatograph, obtain the chromatogram area ratios, measure the peak areas, and calculate the area ratios, as with the Standard solutions. Record from the curves of the Standard solutions the quantities, in mg, of atropine and scopolamine in the weight of specimen taken. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701: 30 min UNIFORMITY OF DOSAGE UNITS 905:

allow the layers to separate, and discard the chloroform layer. [NOTEIf emulsions are formed, a mixed solvent consisting of chloroform-isopropyl alcohol (10:3) may be substituted for chloroform throughout the procedure.] Add another 15 mL of chloroform and extract again, discarding the chloroform phase. Add 15 mL of Phosphate buffer and sufficient 1 N sodium hydroxide to yield a final pH of 9.09.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1 mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Extraction blank: Place 10 mL of Diluent in a 60-mL separator. Proceed as directed under Sample solution, beginning with then add 15 mL of chloroform. The blank chromatogram contains no significant interferences at the locus of atropine, scopolamine, or homatropine. Sample solution: Moisten 10 g, previously reduced to a moderately coarse powder with a mixture of 8 mL of ammonium hydroxide, 10 mL of alcohol, and 20 mL of ether, and extract the alkaloids by either Method I or Method II below. If necessary, reduce the volume of the extract to 100 mL by evaporation on a steam bath. Method I: Place the moistened drug in a continuousextraction thimble, and allow maceration to proceed overnight, then extract with ether for 3 h, or longer if necessary to effect complete extraction. Method II: Place the moistened drug in a small percolator, and allow maceration to proceed overnight. Percolate slowly with a mixture of 3 volumes of ether and 1 volume of chloroform. Continue the percolation until the residue from 3 to 4 mL of percolate last passed, when dissolved in dilute sulfuric acid (1 in 70) and treated with mercuric iodide TS, shows not more than a faint turbidity. Transfer the extract to a separator with the aid of ether. Extract with five 15-mL portions of dilute sulfuric acid (1 in 70), filtering each portion drawn off into a 100-mL volumetric flask. Wash the filter with dilute sulfuric acid (1 in 70), and collect the washings in the flask. Add dilute sulfuric acid (1 in 70) to volume, and mix. Dilute 20.0 mL of the resulting solution with the same dilute acid to 100.0 mL. Pipet 2 mL of Tincture into a 60-mL separator containing 10 mL of Diluent. Add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake vigorously, allow the layers to separate, and discard the chloroform layer. [NOTEIf emulsions are formed, a mixed solvent consisting of chloroform-isopropyl alcohol (10:3) may be substituted for chloroform throughout the extraction procedure.] Add another 15 mL of chloroform, and extract again, discarding the chloroform phase. Add 15 mL of Phosphate buffer and sufficient 1 N sodium hydroxide to yield a final pH of 9.09.5. Add 15 mL of chloroform, shake vigorously, and allow the layers to separate. Filter the organic phase through 10 g of anhydrous sodium sulfate (see Suitability for alkaloid assays under Reagents, Indicators, and SolutionsSodium Sulfate, Anhydrous) previously washed with chloroform and supported in a funnel with a small pledget of glass wool, into a suitable
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS USP Homatropine Hydrobromide RS USP Scopolamine Hydrobromide RS
Belladonna Tincture
(Comment on this Monograph)id=m7440=Belladonna Tincture=B-Monos.pdf) DEFINITION Belladonna Tincture yields, from each 100 mL, NLT 27 mg and NMT 33 mg of the alkaloids of belladonna leaf.
Belladonna Leaf, in moderately coarse powder To make 100 g 1000 mL
Prepare a tincture by Process P as modified for assayed Tinctures (see Pharmaceutical Dosage Forms 1151), using a mixture of 3 volumes of alcohol and 1 volume of water as the menstruum. Finally adjust the Tincture to contain, in each 100 mL, 30 mg of the alkaloids of belladonna leaf. ASSAY PROCEDURE Phosphate buffer: 34.8 g of dibasic potassium phosphate in 900 mL of water Adjust to a pH of 9.5 by the addition of 3 N hydrochloric acid or sodium hydroxide, with mixing. Diluent: Dilute sulfuric acid (1 in 350) Internal standard solution: 0.8 mg/mL of USP Homatropine Hydrobromide RS in Diluent [NOTEPrepare fresh on the day of use.] Standard stock solution A: 1.0 mg/mL of USP Scopolamine Hydrobromide RS in Diluent Standard stock solution B: Dissolve 20 mg of USP Atropine Sulfate RS in 25 mL of Diluent in a 50-mL volumetric flask. Add 2.0 mL of Standard stock solution A. Add Diluent to volume. [NOTEPrepare fresh on the day of use.] Standard solutions: Pipet into three separate 60-mL separators 1.0-, 2.0-, and 3.0-mL portions, respectively, of Standard stock solution A, and add 9.0, 8.0, and 7.0 mL, respectively, of Diluent. Add 1.0 mL of Internal standard solution, then add 15 mL of chloroform, shake vigorously,
24

USP REFERENCE STANDARDS 11 USP Atropine Sulfate RS USP Homatropine Hydrobromide RS USP Scopolamine Hydrobromide RS
USP 32
container. Extract again with two 15-mL portions of chloroform, again collecting the clarified organic phase. Wash the sodium sulfate and the tip of the funnel with 5 mL of chloroform. Evaporate the combined organic phases under reduced pressure, at a temperature below 45, add 1 mL of chloroform, and mix to dissolve the alkaloids, taking care to wet the sides of the container. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.2-m 4-mm; glass column packed with 3% G3 on S1AB [NOTEThe column may be cured and conditioned as specified under Chromatography 621, Gas Chromatography.] Temperature Column: 215 Injector port: 240 Detector: 240 Carrier gas: Dry helium Flow rate: 65 mL/min Injection size: 5 L System suitability Sample: Sample solution for 610 injections Suitability requirements Resolution: NLT 3.0 between aH and aA (R) Tailing factor NMT 2.0, (the sum of the distances from peak center to the leading edge and to the tailing edge divided by twice the distance from peak center to the leading edge) measured at 5% of the peak height of aA Relative standard deviation: The analytical system is suitable for conducting this Assay if the relative standard deviation for the ratio, RA, is NMT 2.0%, calculated by the formula: (standard deviation/mean ratio) 100 Analysis Samples: Standard solutions and Sample solution Measure the areas, aA, aH, and aS, of the atropine, homatropine, and scopolamine peaks, respectively, in each chromatogram, and calculate the ratios AA and AS: Result = aA/aH and aS/aH Plot the Standard curves of the values of RA and RS against the amounts, in mg, of atropine and scopolamine in the solutions. (The ratio of the molecular weight of atropine to that of anhydrous atropine sulfate is 0.8551, and the ratio of the molecular weight of scopolamine to that of anhydrous scopolamine hydrobromide is 0.7894.) Inject a portion of the Sample solution into the chromatograph, obtain the chromatogram area ratios, measure the peak areas, and calculate the area ratios, as with the Standard solutions. Record from the Standard curve the quantities, in mg, of atropine and scopolamine in the specimen. Add the quantity, in mg, of atropine and scopolamine, and multiply by 50 to obtain the weight, in mg, of alkaloids/ 100 mL. Acceptance criteria: 2733 mg of the alkaloids of belladonna leaf/100 g OTHER COMPONENTS ALCOHOL DETERMINATION, Method II 611: 65.0%70.0% of C2H5OH, determined by the gas-liquid chromatographic procedure, acetone being used as the internal standard ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and avoid exposure to direct sunlight and to excessive heat.
Benazepril Hydrochloride
(Comment on this Monograph)id=m7490=Benazepril Hydrochloride=B-Monos.pdf)
460.95 C24H28N2O5 HCl 1H-1-Benzazepine-1-acetic acid, 3-[[1-(ethoxycarbonyl)-3phenylpropyl]amino]-2,3,4,5-tetrahydro-2-oxo-, monohydrochloride, [S-(R*,R*)]-; (3S)-3-[[(1S)-1-Carboxy-3-phenylpropyl]amino]-2,3,4,5tetrahydro-2-oxo-1H-1-benzazepine-1-acetic acid, 3-ethyl ester, monohydrochloride [86541-74-4]. DEFINITION Benazepril Hydrochloride contains NLT 98.0% and NMT 102.0% of C24H28N2O5 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Solution A: 0.81 g of tetrabutylammonium bromide in 360 mL of water containing 0.2 mL of glacial acetic acid Mobile phase: Methanol and Solution A (16:9) System suitability solution: 0.4 mg/mL each of USP Benazepril Hydrochloride RS and USP Benazepril Related Compound B RS in Mobile phase Standard solution: 0.2 mg/mL of USP Benazepril Hydrochloride RS in Mobile phase Sample solution: Transfer about 10.0 mL of the Sample solution (from either Procedure 1 or Procedure 2), prepared as directed in the tests for Impurities, Organic Impurities to a 50mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 3.9-mm 30-cm; packing L1 Guard column: 4.6-mm 3-cm; packing L1 Flow rate: 1 mL/min Injection size: 25 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 1.7 between benazepril hydrochloride and benazepril related compound B Relative standard deviation: NMT 2.0% for both benazepril hydrochloride and benazepril related compound B
USP 32
Analysis Samples: Sample solution and Standard solution Calculate the percentage of C24H28N2O5 HCl in the portion of Benazepril Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benazepril Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of benazepril in the CU Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: Ignite at 600. NMT 0.1% residue is found. HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE 1: TEST FOR BENAZEPRIL RELATED COMPOUND A Buffer solution: 9.66 mg/mL of monobasic potassium phosphate and 2.68 mg/mL of dibasic sodium phosphate, heptahydrate Mobile phase: Methanol and Buffer solution (1:4) System suitability solution: 1.0 mg/mL of USP Benazepril Hydrochloride RS and 0.005 mg/mL of USP Benazepril Related Compound A RS in Mobile phase Standard stock solution: 0.05 mg/mL of USP Benazepril Related Compound A RS in Mobile phase Standard solution A: 5 g/mL of USP Benazepril Related Compound A in Mobile phase, from the Standard stock solution Standard solution B: 1 g/mL of USP Benazepril Related Compound A in Mobile phase, from the Standard stock solution Sample solution: 1.0 mg/mL of Benazepril Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.0-mm 10-cm; packing L41 Temperature: 30 Flow rate: 0.9 mL/min Injection size: 50 L System suitability Samples: System suitability solution, Standard solution A, and Standard solution B [NOTEThe relative retention time of benazepril related compound A1 is about 2.3.] Suitability requirements Resolution: NLT 2.0 between benazepril hydrochloride and benazepril related compound A Signal-to-noise ratio: NLT 10:1, Standard solution B Relative standard deviation: NMT 10.0%, Standard solution A Analysis Samples: Sample solution and Standard solution Calculate the percentage of benazepril related compound A in the portion of Benazepril Hydrochloride taken: Result = (rU/rS) (CS/CU) 100
1((3R)-3-[[(1R)-1-(ethoxycarbonyl)-3-phenylpropyl]amino]-2,3,4,5-
Official Monographs / Benazepril 25

= peak response of benazepril related compound A from the Sample solution rS = peak response of benazepril related compound A from the Standard solution = concentration of USP Benazepril Related CS Compound A RS in the Standard solution (mg/mL) CU = nominal concentration of Benazepril Hydrochloride in the Sample solution (mg/mL) Acceptance criteria Individual impurities: NMT 0.1% PROCEDURE 2: TEST FOR BENAZEPRIL RELATED COMPOUNDS B, C, D, E, F, and G Solution A, Mobile phase, System suitability solution, Chromatographic system and System suitability: Proceed as directed in the Assay. Standard solution: 1 g/mL of USP Benazepril Hydrochloride RS, 10 g/mL each of related compound USP Benazepril Related Compound B RS, USP Benazepril Related Compound C RS, USP Benazepril Related Compound D RS, USP Benazepril Related Compound E RS, USP Benazepril Related Compound F RS, and USP Benazepril Related Compound G RS in Mobile phase Sample solution: 1.0 mg/mL of Benazepril Hydrochloride in Mobile phase Analysis Samples: Sample solution and Standard solution Calculate the percentage of benazepril related compounds in the portion of Benazepril Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response for the relevant benazepril related compound from the Sample solution = peak response for the relevant benazepril related rS compound from the Standard solution = concentration of relevant USP Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of Benazepril CU Hydrochloride in the Sample solution (mg/mL) Acceptance criteria Individual impurities: See Impurity Table 1 [NOTEIn addition to not exceeding the limits for benazepril related compounds in Impurity Table 1, NMT 0.1% of any other single impurity is found.] For calculating any other single unspecified impurity: = concentration of the USP Benazepril CS Hydrochloride RS in the Standard solution Total impurities: NMT 2.0% of total impurities (excluding benazepril related compound A from Procedure 1) is found.
Impurity Table 1 Relative Retention Time 0.4 0.5 0.6 1.5 1.7 Acceptance Criteria, NMT (%) 0.2 0.2 0.3 0.5 0.2
rU
rU
Name Benazepril Related Compound E1 Benazepril Related Compound F2 Benazepril Related Compound C3 Benazepril Related Compound B4 Benazepril Related Compound D5
tetrahydro-2-oxo-1H-1-benzazepine-1-acetic acid, monohydrochloride.
26
Benazepril / Official Monographs

Impurity Table 1 (continued) Relative Retention Time 2.0 Acceptance Criteria, NMT (%) 0.2
acid.
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B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.81 g of tetrabutylammonium bromide in 360 mL of water containing 0.2 mL of glacial acetic acid Mobile phase: Methanol and Solution A (16:9) System suitability solution: 0.4 mg/mL each of USP Benazepril Hydrochloride RS and USP Benazepril Related Compound B RS in Mobile phase Standard solution: 0.2 mg/mL of USP Benazepril Hydrochloride RS in Mobile phase Sample solution: Finely powder NLT 20 Tablets, and transfer a portion of the powder, equivalent to 50 mg of benazepril hydrochloride, to a 250-mL volumetric flask. Add about 150 mL of Mobile phase, and shake by mechanical means for 30 min. Dilute with Mobile phase to volume, mix, and centrifuge. Pass an aliquot of the supernatant through a suitable filter, discarding the first 6 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 3.9-mm 30-cm; packing L1 Guard column: 4.6-mm 3-cm; packing L1 Flow rate: 1 mL/min Injection size: 25 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 1.7 between benazepril hydrochloride and benazepril related compound B Relative standard deviation: NMT 2.0% for each from benazepril hydrochloride and benazepril related compound B Analysis Samples: Standard solution and Sample solution Calculate the percentage of C24H28N2O5 HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benazepril Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of benazepril CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 500 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of C24H28N2O5 HCl dissolved by employing the following method. Solution A, Mobile phase, System suitability solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Analysis: Inject 60 L, or an amount of a filtered portion of the solution under test, equivalent to 1.2 g of benazepril, into the chromatograph. The amount of benazepril injected should not exceed 1.5 g. Record the chromatogram, and measure the responses for the major peaks. Determine the percentage of C24H28N2O5 HCl dissolved in comparison with a Standard solution having a known concentration of USP Benazepril Hydrochloride RS in the same Medium and similarly chromatographed. rU rS CS
Name Benazepril Related Compound G6
13-Amino-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine-1-acetic
2t-Butyl-3-amino-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine-1-acetic
acid. 33-(1-Carboxy-3-phenyl-(1S)-propyl)amino-2,3,4,5-tetrahydro-2-oxo-1H-1(3S)-benzazepine)-1-acetic acid. 4Mixture of diastereoisomers (3-(1-ethoxycarbonyl-3-phenyl-(1R)propyl)amino-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine)-1-acetic acid and (3-(1-ethoxycarbonyl-3-phenyl-(1S)-propyl)amino-2,3,4,5tetrahydro-2-oxo-1H-1-(3R)-benzazepine)-1-acetic acid. 53-(1-Ethoxycarbonyl-3-cyclohexyl-(1S)-propyl)amino-2,3,4,5tetrahydro-2-oxo-1H-1-(3S)-benzazepine)-1-acetic acid monohydrochloride. 63-(1-Ethoxycarbonyl-3-phenyl-(1S)-propyl)amino-2,3,4,5-tetrahydro-2oxo-1H-1-(3S)-benzazepine)-1-acetic acid ethyl ester.
SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.5% of its weight. ABSORBANCE OF SOLUTION: The absorbance of a solution (1 in 100) of it in methanol, determined in a 1-cm cell at 420 nm, is NMT 0.015, methanol being used as the blank. ABSORPTIVITY Sample solution: 0.025 mg/mL of Benazepril Hydrochloride in methanol Analysis: Proceed as directed under Spectrophotometry and Light-Scattering 851, and measure the absorbance at 238 nm. Acceptance criteria: 21.023.2 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store at a temperature below 30, preferably between 15 and 30. USP REFERENCE STANDARDS 11 USP Benazepril Hydrochloride RS USP Benazepril Related Compound A RS USP Benazepril Related Compound B RS USP Benazepril Related Compound C RS USP Benazepril Related Compound D RS USP Benazepril Related Compound E RS USP Benazepril Related Compound F RS USP Benazepril Related Compound G RS
Benazepril Hydrochloride Tablets

(Comment on this Monograph)id=m7495=Benazepril Hydrochloride Tablets=B-Monos.pdf) DEFINITION Benazepril Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of benazepril hydrochloride (C24H28N2O5 HCl). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Equivalent to 1.0 mg/mL of benazepril hydrochloride from powdered Tablets (NLT 20) in methanol [NOTEShake by mechanical means for 15 min. Dilute with methanol to volume, mix, and centrifuge. Pass an aliquot of the supernatant through a suitable filter, discarding the first 6 mL of the filtrate.] Application volume: 20 L Developing solvent system: Ethyl acetate, methanol, and ammonium hydroxide (80:20:15)
USP 32
Tolerances: NLT 80% (Q) of the labeled amount of C24H28N2O5 HCl UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Solution A, Mobile phase, System suitability solution, Standard solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Sample solution: Transfer 1 Tablet to a suitable volumetric flask, add a volume of Mobile phase equivalent to 50% of the volume of the flask, sonicate for 5 min, and then shake by mechanical means for NLT 10 min. Dilute quantitatively, and stepwise if necessary, with Mobile phase to obtain a final concentration of nominally 0.2 mg/mL, mix, and pass a portion of the solution through a suitable filter, discarding the first 6 mL of the filtrate. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C24H28N2O5 HCl in the Tablet: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benazepril Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of benazepril hydrochloride in the Sample solution (mg/mL)
Official Monographs / Bendroflumethiazide 27

Acceptance criteria Individual impurities: NMT 3.0% of benazepril related compound C is found; NMT 1.0% of any other individual impurity is found Total impurities: NMT 2.0% (excluding benazepril related compound C) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Benazepril Hydrochloride RS USP Benazepril Related Compound B RS USP Benazepril Related Compound C RS
Bendroflumethiazide
(Comment on this Monograph)id=m7520=Bendroflumethiazide=B-Monos.pdf)
IMPURITIES Organic Impurities PROCEDURE Solution A, Mobile phase, System suitability solution, and System suitability: Proceed as directed in the Assay. Standard solution: 0.006 mg/mL of USP Benazepril Related Compound C RS in Mobile phase Sample solution: Use the Sample solution as directed in the Assay. Chromatographic system (See Chromatography 621, System Suitability.) Proceed as directed under Assay, except use following Injection size. Injection size: 80 L Analysis Samples: Standard solution and Sample solution Record the chromatograms, and measure the responses of the peaks for benazepril related compound C. Calculate the percentage of benazepril related compound C in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benazepril Related Compound C RS in the Standard solution (mg/mL) = nominal concentration of benazepril CU hydrochloride in the Sample solution (mg/mL) Calculate the percentage of each impurity (other than benazepril related compound C) in the portion of Tablets: rU rS CS Result = (ru/rT) 100 ru rT = peak response for each impurity from the Sample solution = sum of the responses of all the peaks (including benazepril related compound C) from the Sample solution
421.42 C15H14F3N3O4S2 2H-1,2,4-Benzothiadiazine-7-sulfonamide, 3,4-dihydro- 3(phenylmethyl)-6-(trifluoromethyl)-, 1,1-dioxide, ()-; ()-3-Benzyl-3,4-dihydro-6-(trifluoromethyl)-2H-1,2,4benzothiadiazine-7-sulfonamide 1,1-dioxide [73-48-3]. DEFINITION Bendroflumethiazide contains NLT 98.0% and NMT 102.0% of C15H14F3N3O4S2, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K: Previously dried over silica gel for 4 h B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 271 nm Solution: 10 g/mL in methanol Acceptance criteria: Absorptivities, calculated on the anhydrous basis, do not differ by more than 4.0%. C. PROCEDURE Sample solution: Mix 5 mL of dilute hydrochloric acid (1 in 2) with 20 mg of Bendroflumethiazide, boil gently for 1 min, and cool in an ice bath. Analysis: To the Sample solution, add in succession, 0.5 mL of sodium nitrite solution (1 in 1000), 0.5 mL of ammonium sulfamate solution (1 in 200), and 0.5 mL of N-(1naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: A deep red color is produced. ASSAY PROCEDURE Sample: 190 mg of Bendroflumethiazide Analysis: Dissolve the Sample in 80 mL of pyridine in a 250mL tall-form beaker in a well-ventilated hood. Add 3 drops of a saturated solution of azo-violet in methanol, cover the beaker, and gently bubble nitrogen through the solution for 5 min, being careful to avoid any contact between the solution and the cover. Raise the nitrogen delivery tube above the solution surface and, maintaining a gentle flushing with nitrogen and stirring with a magnetic or mechanical stirring device, add 0.1 N sodium methoxide VS from a 10-mL buret inserted through an opening in the
28
Bendroflumethiazide / Official Monographs

USP 2,4-Disulfamyl-5-trifluoromethylaniline RS
USP 32
cover. Titrate to a blue endpoint, approaching the endpoint at a rate of 1 or 2 drops/s. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N sodium methoxide is equivalent to 21.07 mg of C15H14F3N3O4S2. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% SELENIUM 291: The absorbance from the Sample solution prepared with 100 mg of Bendroflumethiazide and 100 mg of magnesium oxide, is NMT one-half that from the Standard solution (NMT 30 ppm). HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities LIMIT OF 2,4-DISULFAMYL-5-TRIFLUOROMETHYLANILINE [NOTEUse low-actinic glassware for the Sample solution and the Standard solution.] Mobile phase: Dissolve 5.62 g of sodium chloride and 1.97 g of anhydrous sodium sulfate in 1000 mL of water in a 2-L volumetric flask. Add 4.0 mL of glacial acetic acid and 800 mL of methanol, and dilute with water to volume. Standard solution: 0.75 g/mL of USP 2,4-Disulfamyl-5trifluoromethylaniline RS in methanol Sample solution: 50 g/mL of Bendroflumethiazide in methanol Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 4.6-mm 30-cm; packing L11 Temperature: 35 5 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.4 between the methanol and 2,4disulfamyl-5-trifluoromethylaniline Relative standard deviation: NMT 3.0% for five replicate injections Analysis Samples: Sample solution and Standard solution Calculate the percentage of 2,4-disulfamyl-5trifluoromethylaniline in the portion of Bendroflumethiazide taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP 2,4-Disulfamyl-5trifluoromethylaniline RS in the Standard solution (g/mL) CU = concentration of bendroflumethiazide in the Sample solution (g/mL) Acceptance criteria: NMT 1.5% rU rS CS SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5%
Bendroflumethiazide Tablets
(Comment on this Monograph)id=m7550=Bendroflumethiazide Tablets=B-Monos.pdf) DEFINITION Bendroflumethiazide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C15H14F3N3O4S2. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEUse low-actinic glassware for the Sample solution and the Standard solution.] Mobile phase: Dissolve 5.62 g of sodium chloride and 1.97 g of anhydrous sodium sulfate in 1000 mL of water in a 2-L volumetric flask. Add 4.0 mL of glacial acetic acid and 800 mL of methanol, and dilute with water to volume. Standard solution: 50 g/mL of USP Bendroflumethiazide RS in methanol Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, nominally equivalent to about 5 mg of bendroflumethiazide, to a 100-mL volumetric flask, add about 70 mL of methanol, and sonicate for 15 min, with occasional shaking. Dilute with methanol to volume, mix, and centrifuge a portion of the solution for 15 min. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 4.6-mm 30-cm; packing L11 Temperature: 35 5 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 3.0% for five replicate injections Analysis Samples: Sample solution and Standard solution Calculate the percentage of C15H14F3N3O4S2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bendroflumethiazide RS in the Standard solution (g/mL) = nominal concentration of bendroflumethiazide in the Sample solution (mg/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Bendroflumethiazide RS
USP 32
Official Monographs / Benoxinate 29

PERFORMANCE TESTS DISSOLUTION 711 [NOTEProtect solutions from light throughout this test.] Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 45 min Detector: UV 271 nm Sample solutions: Sample per Dissolution 711 Standard solution: USP Bendroflumethiazide RS in Medium Analysis: Determine the amount of C15H14F3N3O4S2 dissolved by employing UV absorption on filtered portions of the Sample solution, suitably diluted with water, if necessary, in comparison with a Standard solution having a known concentration of USP Bendroflumethiazide RS. Tolerances: NLT 75% (Q) of the labeled amount of C15H14F3N3O4S2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Bendroflumethiazide RS
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities ORDINARY IMPURITIES 466 Sample solution: Methanol Standard solution: Methanol Eluant: A mixture of chloroform, cyclohexane, and diethylamine (5:4:1) Visualization: 12 SPECIFIC TESTS PH 791: 5.06.0, in a solution (1 in 100) LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Benoxinate Hydrochloride RS
Benoxinate Hydrochloride
(Comment on this Monograph)id=m7600=Benoxinate Hydrochloride=B-Monos.pdf)
Benoxinate Hydrochloride Ophthalmic Solution

(Comment on this Monograph)id=m7630=Benoxinate Hydrochloride Ophthalmic Solution=B-Monos.pdf) DEFINITION Benoxinate Hydrochloride Ophthalmic Solution is a sterile solution of Benoxinate Hydrochloride in water. It contains NLT 95.0% and NMT 105.0% of the labeled amount of C17H28N2O3 HCl. IDENTIFICATION IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Sample solution: Equivalent to 50 mg of benoxinate hydrochloride from a volume of Solution in 25 mL 0.01 N hydrochloric acid Analysis: Proceed as directed under IdentificationOrganic Nitrogenous Bases 181, beginning with Transfer the liquid to a separator. Acceptance criteria: The solution meets the requirements of the test. ASSAY PROCEDURE Standard solution: 400 g/mL of USP Benoxinate Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: Transfer the equivalent of 20 mg of benoxinate hydrochloride from a volume of Ophthalmic Solution, to a separator containing 15 mL of water, add 1 mL of ammonium hydroxide, and extract with five 20-mL portions of ether. Wash the combined ether extracts with 10 mL of water, extract the water washing with 10 mL of ether, and add this ether extract to the main extract. Extract the ether solution with three 5-mL portions of 0.1 N hydrochloric acid, collect the acid extracts in a 50-mL volumetric flask, and dilute with 0.1 N hydrochloric acid to volume. Blank: 0.1 N hydrochloric acid Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.)
C17H28N2O3 HCl 344.88 Benzoic acid, 4-amino-3-butoxy-, 2-(diethylamino)ethyl ester, monohydrochloride; 2-(Diethylamino)ethyl 4-amino-3-butoxybenzoate monohydrochloride [5987-82-6]. DEFINITION Benoxinate Hydrochloride contains NLT 98.5% and NMT 101.5% of C17H28N2O3 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K: Previously dried B. ULTRAVIOLET ABSORPTION 197U Sample solution: 15 g/mL in water C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements of the tests Sample solution: 10 mg/mL ASSAY PROCEDURE Sample solution: Dissolve 250 mg of Benoxinate Hydrochloride in a mixture of 20 mL of glacial acetic acid and 20 mL of acetic anhydride. Analysis: Titrate with 0.1 N perchloric acid VS, determining the endpoint potentiometrically. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 34.49 mg of C17H28N2O3 HCl.
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Benoxinate / Official Monographs
USP 32
mixture for 10 min. Cool, add 0.2 g of sodium nitrite to 1 mL of the clear liquid, and add this mixture to 20 mg of naphthol dipotassium disulfonate or naphthol disodium disulfonate in 1 mL of ammonium hydroxide. Acceptance criteria: The solution turns orange-red, and a brown precipitate may be formed. ASSAY PROCEDURE Sample: 0.3 g of Benzethonium Chloride Analysis: Dissolve the Sample in 75 mL of water contained in a glass-stoppered, 250-mL flask. Add 0.4 mL of bromophenol blue solution (1 in 2000), 10 mL of chloroform, and 1 mL of 1 N sodium hydroxide. Titrate with 0.02 M sodium tetraphenylboron VS until the blue color disappears from the chloroform layer. Add the last portions of the sodium tetraphenylboron solution dropwise, agitating vigorously after each addition. Each mL of 0.02 M sodium tetraphenylboron is equivalent to 8.962 mg of C27H42ClNO2. Acceptance criteria: 97.0%103.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities LIMIT OF AMMONIUM COMPOUNDS: To 5 mL of a solution (1 in 50), add 3 mL of 1 N sodium hydroxide, and heat to boiling: the odor of ammonia is not perceptible. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 158163, the specimen having been dried previously LOSS ON DRYING 731: Dry at 105 for 4 h: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.
Mode: UV Analytical wavelength: 308 nm Cell: 1 cm Analysis Samples: Blank, Sample solution, and Standard solution Transfer 5.0 mL each of the Standard solution, Sample solution, and Blank, to separate 200-mL volumetric flasks. Dilute the contents of each flask with water to volume. Concomitantly determine the absorbances of the solutions, using the Blank to set the instrument. Calculate the percentage of C17H28N2O3 HCl in each mL of the Ophthalmic Solution: Result = (AU/AS) (CS/CU) 100 = absorbance from the Sample solution = absorbance from the Standard solution = concentration of USP Benoxinate Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of benoxinate CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% SPECIFIC TESTS STERILITY TESTS 71: PH 791: 3.06.0 Meets the requirements AU AS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Benoxinate Hydrochloride RS
Benzethonium Chloride
(Comment on this Monograph)id=m7960=Benzethonium Chloride=B-Monos.pdf)
Benzethonium Chloride Concentrate

(Comment on this Monograph)id=m7965=Benzethonium Chloride Concentrate=B-Monos.pdf) 448.08 C27H42ClNO2 Benzenemethanaminium, N,N-dimethyl-N-[2-[2-[4-(1,1,3,3tetramethylbutyl)phenoxy]ethoxy]ethyl]-, chloride; Benzyldimethyl[2-[2-[p-(1,1,3,3tetramethylbutyl)phenoxy]ethoxy]ethyl]ammonium chloride [121-54-0]. DEFINITION Benzethonium Chloride contains NLT 97.0% and NMT 103.0% of C27H42ClNO2, calculated on the dried basis. IDENTIFICATION A. PROCEDURE Sample solution: 10 mg/mL Analysis: To 1 mL of the Sample solution, add 2 mL of alcohol, 0.5 mL of 2 N nitric acid, and 1 mL of silver nitrate TS. Acceptance criteria: A white precipitate, which is insoluble in 2 N nitric acid but soluble in 6 N ammonium hydroxide, is formed. B. A solution (1 in 100) forms precipitates with 2 N nitric acid and with mercuric chloride TS, both of which dissolve upon the addition of alcohol. C. PROCEDURE Sample solution: 0.1 g in 1 mL of sulfuric acid Analysis: Add 0.1 g of potassium nitrate, and heat on a steam bath for 3 min. Cautiously dilute the solution with water to 10 mL, add 0.5 g of granulated zinc, and warm the DEFINITION Benzethonium Chloride Concentrate contains NLT 94.0% and NMT 106.0% of the labeled amount of benzethonium chloride (C27H42ClNO2). IDENTIFICATION A. PROCEDURE Sample solution: Evaporate a volume of Concentrate, equivalent to 200 mg of benzethonium chloride, on a steam bath. Analysis: To the residue, add 2 mL of alcohol, 0.5 mL of 2 N nitric acid, and 1 mL of silver nitrate TS. Acceptance criteria: A white precipitate, which is insoluble in 2 N nitric acid but soluble in 6 N ammonium hydroxide, is formed. B. PROCEDURE: Evaporate a volume of Concentrate, equivalent to 200 mg of benzethonium chloride, on a steam bath. Acceptance criteria: The residue so obtained forms precipitates with 2 N nitric acid and with mercuric chloride TS, both of which dissolve upon the addition of alcohol. C. PROCEDURE Sample solution: Evaporate a volume of Concentrate, equivalent to 200 mg of benzethonium chloride, on a steam bath. Analysis: To the residue, add 0.1 g of potassium nitrate, and heat on a steam bath for 3 min. Cautiously dilute the solution with water to 10 mL, add 0.5 g of granulated zinc,
USP 32
and warm the mixture for 10 min. Cool, add 0.2 g of sodium nitrite to 1 mL of the clear liquid, and add this mixture to 20 mg of naphthol dipotassium disulfonate or naphthol disodium disulfonate in 1 mL of ammonium hydroxide. Acceptance criteria: The solution turns orange-red, and a brown precipitate may be formed. ASSAY PROCEDURE Sample solution: Equivalent to 200 mg of benzethonium chloride from a volume of Concentrate, in a glass-stoppered flask Analysis: Add 0.4 mL of bromophenol blue solution (1 in 2000), 10 mL of chloroform, and 1 mL of 1 N sodium hydroxide. Titrate with 0.02 M sodium tetraphenylboron VS until the blue color disappears from the chloroform layer. Add the last portions of the sodium tetraphenylboron solution dropwise, agitating vigorously after each addition. Each mL of 0.02 M sodium tetraphenylboron is equivalent to 8.962 mg of C27H42ClNO2. Acceptance criteria: 94.0%106.0% IMPURITIES Inorganic Impurities LIMIT OF NITRITES: To 1 drop of Concentrate on a spot plate, add 1 drop each of glacial acetic acid, sulfanilic acid in acetic acid solution (1 in 100), and 1-naphthylamine-acetic acid solution (prepared by boiling 30 mg of 1naphthylamine in 70 mL of water, decanting the colorless solution from the blue-violet residue, and mixing with 30 mL of glacial acetic acid): no red color develops in the resulting solution within 10 min. SPECIFIC TESTS OXIDIZING SUBSTANCES: To 5 mL of Concentrate, add 0.5 mL of potassium iodide TS and a few drops of 3 N hydrochloric acid: the solution does not acquire a yellow color. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at room temperature. LABELING: The label states that this article is not intended for direct administration to humans or animals.
Official Monographs / Benzethonium 31

Acceptance criteria: The residue so obtained, forms precipitates with 2 N nitric acid and with mercuric chloride TS, both of which dissolve upon the addition of alcohol. C. PROCEDURE Sample solution: Evaporate a volume of Topical Solution, equivalent to 200 mg of benzethonium chloride, on a steam bath. Analysis: To the residue, add 0.1 g of potassium nitrate, and heat on a steam bath for 3 min. Cautiously dilute the solution with water to 10 mL, add 0.5 g of granulated zinc, and warm the mixture for 10 min. Cool, add 0.2 g of sodium nitrite to 1 mL of the clear liquid, and add this mixture to 20 mg of naphthol dipotassium disulfonate or naphthol disodium disulfonate in 1 mL of ammonium hydroxide. Acceptance criteria: The solution turns orange-red, and a brown precipitate may be formed. ASSAY PROCEDURE Sample solution: Equivalent to 200 mg of benzethonium chloride from a volume of Topical Solution, in a glassstoppered flask. Analysis: Add 0.4 mL of bromophenol blue solution (1 in 2000), 10 mL of chloroform, and 1 mL of 1 N sodium hydroxide. Titrate with 0.02 M sodium tetraphenylboron VS until the blue color disappears from the chloroform layer. Add the last portions of the sodium tetraphenylboron solution dropwise, agitating vigorously after each addition. Each mL of 0.02 M sodium tetraphenylboron is equivalent to 8.962 mg of C27H42ClNO2. Acceptance criteria: 95.0%105.0% IMPURITIES Inorganic Impurities LIMIT OF NITRITES: To 1 drop of Topical Solution on a spot plate, add 1 drop each of glacial acetic acid, sulfanilic acid in acetic acid (1 in 100), and 1-naphthylamine-acetic acid solution (prepared by boiling 30 mg of 1-naphthylamine in 70 mL of water, decanting the colorless solution from the blue-violet residue, and mixing with 30 mL of glacial acetic acid): no red color develops in the resulting solution within 10 min. SPECIFIC TESTS OXIDIZING SUBSTANCES: To 5 mL, add 0.5 mL of potassium iodide TS and a few drops of 3 N hydrochloric acid: the solution does not acquire a yellow color. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.
Benzethonium Chloride Topical Solution

(Comment on this Monograph)id=m7990=Benzethonium Chloride Topical Solution=B-Monos.pdf) DEFINITION Benzethonium Chloride Topical Solution contains NLT 95.0% and NMT 105.0% of the labeled amount of benzethonium chloride (C27H42ClNO2). IDENTIFICATION A. PROCEDURE Sample solution: Evaporate a volume of Topical Solution, equivalent to 200 mg of benzethonium chloride, on a steam bath. Analysis: To the residue, add 2 mL of alcohol, 0.5 mL of 2 N nitric acid, and 1 mL of silver nitrate TS. Acceptance criteria: A white precipitate, which is insoluble in 2 N nitric acid but soluble in 6 N ammonium hydroxide, is formed. B. Evaporate a volume of Topical Solution, equivalent to 200 mg of benzethonium chloride, on a steam bath.
Benzethonium Chloride Tincture

(Comment on this Monograph)id=m8000=Benzethonium Chloride Tincture=B-Monos.pdf) DEFINITION Benzethonium Chloride Tincture contains, in each 100 mL, NLT 190 mg and NMT 210 mg of C27H42ClNO2. Benzethonium Chloride Tincture is prepared as follows.
Benzethonium Chloride Alcohol 2g 685 mL
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Benzethonium / Official Monographs

100 mL a sufficient quantity 1000 mL
USP 32
Analysis: From the respective chromatograms obtained as described previously, calculate the ratios of peak areas for alcohol to internal standard and for acetone to internal standard. Calculate the percentage of alcohol and of acetone in the portion of Tincture taken: Result = [A(Y Z) + B(Z X)]/(Y X) = percentage of alcohol, or of acetone, in the lower standards B = percentage of alcohol, or of acetone, in the higher standards X = ratio of the alcohol peak areas, or the acetone peak areas, to the internal standard peak areas for the lower standard Y = ratio of the alcohol peak areas, or the acetone peak areas, to the internal standard peak areas for the higher standard Z = ratio of the alcohol peak areas, or the acetone peak areas, to the internal standard peak areas for the Sample solution Acceptance criteria C2H5OH: 62.0%68.0% C3H6O: 9.0%11.0% SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.8680.876 A
Acetone Purified Water To make
Dissolve the Benzethonium Chloride in a mixture of the Alcohol and the Acetone. Add sufficient Purified Water to make 1000 mL. [NOTEBenzethonium Chloride Tincture may be colored by the addition of any suitable color or combination of colors certified by the FDA for use in drugs.] IDENTIFICATION A. PROCEDURE Sample: 50 mL Analysis: To the residue obtained by evaporating the Sample on a steam bath, add 2 mL of alcohol, 0.5 mL of 2 N nitric acid, and 1 mL of silver nitrate TS. Acceptance criteria: A white precipitate, which is insoluble in 2 N nitric acid but soluble in 6 N ammonium hydroxide, is formed. B. Evaporate 50 mL on a steam bath. Acceptance criteria: The residue obtained forms precipitates with 2 N nitric acid and with mercuric chloride TS, both of which dissolve upon the addition of alcohol. ASSAY PROCEDURE Sample solution: Place 50.0 mL of Tincture in a 150-mL beaker, and add, with continuous stirring, 10 mL of sodium tetraphenylboron solution (1 in 40). Cover, and allow to stand for 16 h. Decant the supernatant into a tared sinteredglass crucible, applying vacuum filtration. Suspend the precipitate in 20 mL of water. Analysis: Transfer the precipitate to the crucible, washing well with water. Dry the precipitate and the crucible at 105 for 1 h, cool, and weigh. The weight of the precipitate so obtained, multiplied by 0.6122, represents its equivalent of C27H42ClNO2. Acceptance criteria: 190210 mg OTHER COMPONENTS ALCOHOL AND ACETONE CONTENT Standard solution A: Dehydrated alcohol, methanol as the internal standard, and water (11:5:84) Standard solution B: Dehydrated alcohol, methanol as the internal standard, and water (14:5:81) Standard solution C: Acetone, methanol as the internal standard, and water (1.7:5:93.3) Standard solution D: Acetone, methanol as the internal standard, and water (2.2:5:92.8) Sample solution: Benzethonium Chloride Tincture, methanol as the internal standard, and water (4:1:15) Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 120-cm 4-mm column packed with a suitable type of support, such as 80- to 100-mesh S3 Carrier gas: Dry helium Temperatures Column: 120 Injection port: 240 Detector block: 240 Flow rate: 90 mL/min Injection size: 0.8 L Analysis Samples: Standard solutions A, B, C, and D and Sample solution
Benzocaine
(Comment on this Monograph)id=m8050=Benzocaine=BMonos.pdf)
C9H11NO2 Benzoic acid, 4-amino-, ethyl ester; Ethyl p-aminobenzoate [94-09-7].
165.19
DEFINITION Benzocaine, dried over phosphorus pentoxide for 3 h, contains NLT 98.0% and NMT 102.0% of C9H11NO2. IDENTIFICATION A. INFRARED ABSORPTION 197K: Previously dried over phosphorus pentoxide for 3 h B. ULTRAVIOLET ABSORPTION 197U Wavelength: 278 nm Solution: 5 g/mL in chloroform Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0%. C. PROCEDURE Sample solution: 20 mg in 10 mL of water with the aid of a few drops of 3 N hydrochloric acid Analysis: Add 5 drops of a solution of sodium nitrite (1 in 10), followed by 2 mL of a solution of 100 mg of 2naphthol in 5 mL of 1 N sodium hydroxide.
USP 32
Acceptance criteria: An orange-red precipitate is formed.
Official Monographs / Benzocaine 33

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Benzocaine RS
ASSAY PROCEDURE Solution A: Acetic acid, triethylamine, and water (20:1:980) The pH should be between 2.953.0 (adjust as needed). Mobile phase: Methanol and Solution A (2:3) Standard solution: 0.024 mg/mL of USP Benzocaine RS in Mobile phase Sample solution: 0.024 mg/mL of Benzocaine in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 285 nm Column: 2.0-mm 15-cm; 5-m packing L11 Flow rate: 0.2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis: Samples: Standard solution and Sample solution Calculate the percentage of C9H11NO2 in the portion taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benzocaine RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE: To a solution of 200 mg in 5 mL of alcohol, previously acidified with a few drops of diluted nitric acid, add a few drops of silver nitrate TS: no turbidity is produced immediately. HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities ORDINARY IMPURITIES 466 Sample solution: Dehydrated alcohol Standard solution: Dehydrated alcohol Application volume: 10 L Eluant: Chloroform containing about 0.75% of dehydrated alcohol as a preservative, in a nonequilibrated chamber Visualization: 1 Limit: The total of any ordinary impurities observed does not exceed 1%. READILY CARBONIZABLE SUBSTANCES TEST 271: Dissolve 500 mg in 5 mL of sulfuric acid TS: the solution has no more color than Matching Fluid A. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 8892, but the range between beginning and end of melting does not exceed 2 REACTION: Dissolve 1.0 g in 10 mL of neutralized alcohol: a clear solution results. Dilute this solution with 10 mL of water, and add 2 drops of phenolphthalein TS and 1 drop of 0.10 N sodium hydroxide: a red color is produced. LOSS ON DRYING 731: Dry over phosphorus pentoxide for 3 h: it loses NMT 1.0% of its weight.
Benzocaine Topical Aerosol

(Comment on this Monograph)id=m8060=Benzocaine Topical Aerosol=B-Monos.pdf) DEFINITION Benzocaine Topical Aerosol is a solution of Benzocaine in a pressurized container. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H11NO2. IDENTIFICATION PROCEDURE Sample solution: Spray a portion of Topical Aerosol, equivalent to 5 mg of benzocaine, into a beaker, and heat on a steam bath for a few min to expel residual propellant. Add 20 mL of 0.5 N hydrochloric acid, warm gently to disperse, cool, and filter. Analysis: To 10 mL of the filtrate, add 5 drops of sodium nitrite solution (1 in 10) and 2 drops of methyl red TS, and neutralize with 1 N sodium hydroxide. Add 2 mL of a solution of 100 mg of 2-naphthol in 5 mL of 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. ASSAY PROCEDURE Sample solution: Spray a portion of the Topical Aerosol into a beaker, and heat on a steam bath for a few minutes to expel residual propellant. Cool, and transfer a portion of the solution, equivalent to 200 mg of benzocaine, to a 250mL beaker. Add 50 mL of water and 5 mL of hydrochloric acid, and stir. Cool the solution in an ice bath to 10. Analysis: Titrate slowly with 0.1 M sodium nitrite VS, using a calomelplatinum electrode system. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 M sodium nitrite is equivalent to 16.52 mg of C9H11NO2. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS OTHER REQUIREMENTS: It meets the requirements for Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers 601, Pressure Test, Minimum Fill, and Leakage Test. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in pressurized containers, and avoid exposure to excessive heat.
Benzocaine Cream
(Comment on this Monograph)id=m8070=Benzocaine Cream=B-Monos.pdf) DEFINITION Benzocaine Cream contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H11NO2 in a suitable cream base. IDENTIFICATION PROCEDURE Sample solution: Place an amount equivalent to 50 mg of benzocaine from Cream in a small beaker, add 20 mL of 0.5
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USP 32
filtrate. Transfer 1.0 mL of the clear solution to a second 100-mL volumetric flask, and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 294 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C9H11NO2 in the portion of Gel taken: Result = (rU/rS) (CS/CU) 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Benzocaine RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Benzocaine RS rU rS CS
N hydrochloric acid, warm gently to disperse the Cream, cool, and filter. Analysis: To 10 mL of the filtrate add 5 drops of a solution of sodium nitrite (1 in 10) and 2 drops of methyl red TS, and neutralize with 1 N sodium hydroxide. Add 2 mL of a 20 mg/mL solution of 2-naphthol in 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. ASSAY PROCEDURE Sample solution: An amount of Cream, nominally equivalent to 200 mg of benzocaine in a tared 250-mL beaker Analysis: Add 50 mL of water and 5 mL of hydrochloric acid, and stir by mechanical means, with gentle warming, until a solution is effected. Cool the solution in an ice bath to about 10. Titrate slowly with 0.1 M sodium nitrite VS, using a calomel-platinum electrode system. Each mL of 0.1 M sodium nitrite is equivalent to 16.52 mg of C9H11NO2. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, and avoid prolonged exposure to temperatures exceeding 30.
Benzocaine Gel
(Comment on this Monograph)id=m8073=Benzocaine Gel=BMonos.pdf) DEFINITION Benzocaine Gel contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H11NO2. IDENTIFICATION A. PROCEDURE Sample solution: Place an amount equivalent to 5 mg of benzocaine from Gel in a beaker, add 20 mL of 0.5 N hydrochloric acid, warm gently to disperse the Gel, cool, and filter, if necessary, to obtain a clear solution. Analysis: To 10 mL of the clear solution, add 5 drops of a solution of sodium nitrite (1 in 10) and 2 drops of methyl red TS, and neutralize with 1 N sodium hydroxide. Add 2 mL of a solution of 100 mg of 2-naphthol in 5 mL of 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol and water (1:1) Mobile phase: Methanol, glacial acetic acid, and water (14:1:10) Standard solution: 0.02 mg/mL of USP Benzocaine RS in Diluent Sample solution: Place an amount equivalent to 200 mg of benzocaine from Gel, in a 100-mL volumetric flask, add 80 mL of Diluent, and shake by mechanical means for 15 min. Dilute with Diluent to volume, and filter, if necessary, to obtain a clear solution, discarding the first 5 mL of the
Benzocaine Lozenges
(Comment on this Monograph)id=m8077=Benzocaine Lozenges=B-Monos.pdf) DEFINITION Benzocaine Lozenges contain NLT 85.0% and NMT 120.0% of the labeled amount of C9H11NO2. IDENTIFICATION A. PROCEDURE Sample solution: Place an amount equivalent to 20 mg of benzocaine from powdered Lozenges, in 10 mL of water with the aid of a few drops of 3 N hydrochloric acid. Filter, if necessary, to obtain a clear solution. Analysis: Add 5 drops of a 100 mg/mL sodium nitrite solution, followed by 2 mL of a 20 mg/mL solution of 2naphthol in 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, 1.0 M monobasic potassium phosphate solution previously adjusted with phosphoric acid to a pH of 3.0, and water (5:1:14) Standard solution A: 0.01 mg/mL of USP Benzocaine RS in 0.1 N hydrochloric acid
USP 32
Standard solution B: 0.01 mg/mL of USP Benzocaine RS in a mixture of acetonitrile and water (1:1) Sample stock solution A: Add the equivalent of 40 mg of benzocaine from powdered Lozenges (NLT 20 Lozenges), to a 200-mL volumetric flask, add 150 mL of 0.1 N hydrochloric acid, and stir for NLT 2 h. Dilute with 0.1 N hydrochloric acid to volume. Sample solution A: Equivalent to 0.02 mg/mL of benzocaine in 0.1 N hydrochloric acid from Sample stock solution A Sample stock solution B: Add the equivalent of 40 mg of benzocaine from powdered Lozenges (NLT 20 Lozenges), to a 200-mL volumetric flask, add 150 mL of a mixture of acetonitrile and water (1:1), and stir for NLT 30 min. Dilute with the mixture of acetonitrile and water (1:1) to volume. Sample solution B: Equivalent to 0.02 mg/mL of benzocaine in a mixture of acetonitrile and water (1:1), from Sample stock solution B Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: Standard solution A and Standard solution B Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution A, Sample solution B, Standard solution A, and Standard solution B Calculate the percentage of total C9H11NO2 taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution A = peak response from the Standard solution A = concentration of USP Benzocaine RS in Standard solution A (mg/mL) = nominal concentration of Sample solution A CU (mg/mL) Calculate the percentage of free C9H11NO2 taken: Result = (rU/rS) (CS/CU) 100 = peak response from Sample solution B = peak response from Standard solution B = concentration of USP Benzocaine RS in Standard solution B (mg/mL) = nominal concentration of Sample solution B CU (mg/mL) Acceptance criteria: 85.0%120.0% rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Benzocaine RS rU rS CS

IDENTIFICATION OINTMENTS HAVING WATER-SOLUBLE BASES Sample solution: Transfer a portion of Ointment, equivalent to 5 mg of benzocaine, to a small beaker. Add 20 mL of 0.5 N hydrochloric acid, warm gently to disperse the Ointment, cool, and filter, if necessary. Analysis: To 10 mL of the filtrate, add 5 drops of a solution of sodium nitrite (1 in 10) and 2 drops of methyl red TS, and neutralize with 1 N sodium hydroxide. Add 2 mL of a 20 mg/mL solution of 2-naphthol in sodium hydroxide TS. Acceptance criteria: An orange-red precipitate is formed. OINTMENTS HAVING WATER-INSOLUBLE BASES Sample solution: Transfer a portion of Ointment, equivalent to 2.5 mg of benzocaine, to a small separator, and dissolve in 20 mL of ether. Extract with 10 mL of 0.5 N hydrochloric acid, and filter the aqueous phase through paper into a small beaker. Analysis: Add 5 drops of a solution of sodium nitrite (1 in 10) and 2 drops of methyl red TS, and neutralize with 1 N sodium hydroxide. Add 2 mL of a 20 mg/mL solution of 2naphthol in 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. ASSAY OINTMENTS HAVING WATER-SOLUBLE BASES Sample solution: Ointment, nominally equivalent to 200 mg of benzocaine, in a tared 250-mL beaker Analysis: Add 50 mL of water and 5 mL of hydrochloric acid, and stir by mechanical means until a solution is effected. Cool the solution in an ice bath to about 10. Titrate slowly with 0.1 M sodium nitrite VS, using a calomelplatinum electrode system. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 M sodium nitrite is equivalent to 16.52 mg of C9H11NO2. Acceptance criteria: 90.0%110.0% OINTMENTS HAVING WATER-INSOLUBLE BASES Sample solution: Ointment, nominally equivalent to 200 mg of benzocaine Analysis: Transfer to a 125-mL separator. Suspend the Ointment in 50 mL of ether by shaking, and extract with three successive 25-mL portions of 1 N hydrochloric acid, filtering each portion into a 250-mL beaker. Cool the solution in an ice bath to about 10. Titrate slowly with 0.1 M sodium nitrite VS, using a calomelplatinum electrode system. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 M sodium nitrite is equivalent to 16.52 mg of C9H11NO2. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, and avoid prolonged exposure to temperatures exceeding 30.
Benzocaine Ointment
(Comment on this Monograph)id=m8080=Benzocaine Ointment=B-Monos.pdf) DEFINITION Benzocaine Ointment contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H11NO2 in a suitable ointment base.
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USP 32
Analysis: Add 50 mL of water and 5 mL of hydrochloric acid, and stir. Cool the solution in an ice bath to about 10. Titrate slowly with 0.1 M sodium nitrite VS using a calomelplatinum electrode system. Perform a blank determination, and make any necessary correction (See Titrimetry 541). Each mL of 0.1 M sodium nitrite is equivalent to 16.52 mg of C9H11NO2. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, and avoid prolonged exposure to temperatures exceeding 30.
Benzocaine Otic Solution

(Comment on this Monograph)id=m8090=Benzocaine Otic Solution=B-Monos.pdf) DEFINITION Benzocaine Otic Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H11NO2. IDENTIFICATION PROCEDURE Sample solution: Place an amount equivalent to 2.5 mg of benzocaine from a volume of Otic Solution in 10 mL of water, and dissolve with the aid of a few drops of 3 N hydrochloric acid. Analysis: Add 5 drops of a sodium nitrite solution (1 in 10), then add 2 mL of a 20 mg/mL solution of 2-naphthol in 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. ASSAY PROCEDURE Sample: A volume of Otic Solution nominally equivalent to 400 mg of benzocaine Analysis: Add 150 mL of cold 1.6 N hydrochloric acid, and stir until a fine dispersion is obtained. Cool the solution in an ice bath to 10. Titrate slowly with 0.1 M sodium nitrite VS, using a calomelplatinum electrode system. Perform a blank determination, and make any necessary correction (See Titrimetry 541). Each mL of 0.1 M sodium nitrite is equivalent to 16.52 mg of C9H11NO2. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Salmonella species and Escherichia coli and for absence of Staphylococcus aureus and Pseudomonas aeruginosa; the total aerobic microbial count is less than 100 cfu/mL. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.
Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Aerosol

(Comment on this Monograph)id=m8120=Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Aerosol=BMonos.pdf) DEFINITION Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Aerosol is Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Solution packaged in a pressurized container with a suitable inert propellant. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of benzocaine (C9H11NO2), butamben (C11H15NO2), and tetracaine hydrochloride (C15H24N2O2 HCl). IDENTIFICATION The retention times of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol and water (3:2) Mobile phase: Methanol, 0.25 M sodium 1heptanesulfonate, and water (25:1:25) Standard solution: Transfer 140 mg of USP Benzocaine RS to a 100-mL volumetric flask. Swirl with the aid of 25 mL of methanol, and transfer 140 J mg of USP Butamben RS to the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, as a percentage, of butamben to the labeled amount, as a percentage, of benzocaine in the Topical Aerosol. Transfer 140 J mg of USP Tetracaine Hydrochloride RS into the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, as a percentage, of tetracaine hydrochloride to the labeled amount, as a percentage, of benzocaine in the Topical Aerosol. Sonicate for 1 min, and dilute with Diluent to volume. Sample stock solution: Fit the valve of the Topical Aerosol container with a cannula, and expel the contents of the container into a 100-mL, glass-stoppered, round-bottom flask. Attach the flask to a rotary evaporator, and evaporate under a vacuum of 600 mm of mercury for 2.5 h. Transfer a portion of the material in the round-bottom flask, equivalent to 1400 mg of benzocaine, to a 100-mL volumetric flask. Add 75 mL of methanol, and mix. Sonicate for 1 min, dilute with methanol to volume, and mix.
Benzocaine Topical Solution

(Comment on this Monograph)id=m8100=Benzocaine Topical Solution=B-Monos.pdf) DEFINITION Benzocaine Topical Solution is a solution of Benzocaine in a suitable solvent. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H11NO2. It contains a suitable antimicrobial agent. IDENTIFICATION PROCEDURE Sample solution: Transfer an amount of Topical solution, equivalent to 5 mg of benzocaine, to a small beaker and add 20 mL of 0.5 N hydrochloric acid, warm gently to disperse, cool, and filter. Analysis: To 10 mL of the filtrate, add 5 drops of sodium nitrite solution (1 in 10) and 2 drops of methyl red TS, and neutralize with 1 N sodium hydroxide. Add 2 mL of a 20 mg/mL solution of 2-naphthol in 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. ASSAY PROCEDURE Sample solution: Topical Solution equivalent to 200 mg of benzocaine in a 250-mL separator
USP 32
Sample solution: 1.4 mg/mL from the Sample stock solution in Diluent [NOTEPrepare by adding a quantity of Sample stock solution corresponding to 10% of the total volume to a quantity of Diluent corresponding to 75% of the total volume. Shake, and dilute with Diluent to volume.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 313 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for benzocaine and butamben are 0.3 and 0.8, respectively.] Suitability requirements Resolution: NLT 2 between the benzocaine peak and the butamben peak and between the butamben peak and the tetracaine peak Relative standard deviation: NMT 2.0% for each of the three analyte peaks Analysis Samples: Standard solution and Sample solution Calculate the individual percentages of C9H11NO2, C11H15NO2, and C15H24N2O2 HCl in the portion of the residue of Topical Aerosol taken from the round-bottom flask: Result = (RU/RS) (CS/CU) 100 = peak response from the corresponding analyte of the Sample solution = peak response from the corresponding analyte of RS the Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of the individual CU quantities in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS OTHER REQUIREMENTS: It meets the requirements for Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers 601, Pressure Test, Minimum Fill, and Leakage Test. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in pressurized containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Benzocaine RS USP Butamben RS USP Tetracaine Hydrochloride RS

ASSAY PROCEDURE Diluent: Methanol and water (3:2) Mobile phase: Methanol, 0.25 M sodium 1heptanesulfonate, and water (25:1:25) Standard solution: Transfer 140 mg of USP Benzocaine RS, in a 100-mL volumetric flask with the aid of 25 mL of methanol, and swirl. Transfer 140 J mg of USP Butamben RS to the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, in percentage of butamben to the labeled amount, in percentage of benzocaine in the topical solution. Transfer 140 J mg of USP Tetracaine Hydrochloride RS, into the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, in percentage of tetracaine hydrochloride to the labeled amount, in percentage of benzocaine in the topical solution. Sonicate for 1 min, and dilute with Diluent to volume. Sample stock solution: Nominally equivalent to 14 mg/mL of benzocaine from Gel in methanol [NOTESonicate for about 1 min.] Sample solution: Nominally 1.4 mg/mL from the Sample stock solution diluted with Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 313 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for benzocaine, butamben, and tetracaine are about 0.3, 0.8, and 1.0, respectively.] Suitability requirements Resolution: NLT 2 between the benzocaine peak and the butamben peak, and between the butamben peak and the tetracaine peak Relative standard deviation: NMT 2.0% for each of the three analyte peaks Analysis Samples: Sample solution and Standard solution Calculate the individual percentages of C9H11NO2, C11H15NO2, and C15H24N2O2 HCl in the portion of Gel taken: Result = (rU/rS) (CS/CU) 100 = peak area in the corresponding analyte from the Sample solution = peak area in the corresponding analyte from the rS Standard solution = concentration of the appropriate Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of the analyte in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing. USP REFERENCE STANDARDS 11 USP Benzocaine RS USP Butamben RS USP Tetracaine Hydrochloride RS
Benzocaine, Butamben, and Tetracaine Hydrochloride Gel

(Comment on this Monograph)id=m8104=Benzocaine, Butamben, and Tetracaine Hydrochloride Gel=B-Monos.pdf) DEFINITION Benzocaine, Butamben, and Tetracaine Hydrochloride Gel is Benzocaine, Butamben, and Tetracaine Hydrochloride in a suitable gel base. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of benzocaine (C9H11NO2), butamben (C11H15NO2), and tetracaine hydrochloride (C15H24N2O2 HCl). IDENTIFICATION The retention times of the Sample solution correspond to those of the Standard solution, as obtained in the Assay.
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USP 32
Analysis Samples: Sample solution and Standard solution Calculate the individual percentages of C9H11NO2, C11H15NO2, and C15H24N2O2 HCl in the portion of Ointment taken: Result = (rU/rS) (CS/CU) 100 = peak response from the corresponding analyte from the Sample solution = peak response from the corresponding analyte rS from the Standard solution CS = concentration of the appropriate Reference Standard in the Standard solution (mg/mL) = nominal concentration of the analyte in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU SPECIFIC TESTS MINIMUM FILL 755: Meets the requirements
Benzocaine, Butamben, and Tetracaine Hydrochloride Ointment

(Comment on this Monograph)id=m8106=Benzocaine, Butamben, and Tetracaine Hydrochloride Ointment=BMonos.pdf) DEFINITION Benzocaine, Butamben, and Tetracaine Hydrochloride Ointment is Benzocaine, Butamben, and Tetracaine Hydrochloride in a suitable ointment base. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of benzocaine (C9H11NO2), butamben (C11H15NO2), and tetracaine hydrochloride (C15H24N2O2 HCl). IDENTIFICATION The retention times of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol and water (3:2) Mobile phase: Methanol, 0.25 M sodium 1heptanesulfonate, and water (25:1:25) Standard solution: Transfer 140 mg of USP Benzocaine RS, in a 100-mL volumetric flask with the aid of 25 mL of methanol, and swirl. Transfer 140 J mg of USP Butamben RS to the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, as a percentage, of butamben to the labeled amount, as a percentage, of benzocaine in the Ointment. Transfer 140 J mg of USP Tetracaine Hydrochloride RS, into the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, as a percentage, of tetracaine hydrochloride to the labeled amount, as a percentage, of benzocaine in the Ointment. Sonicate for about 1 min, and dilute with Diluent to volume. Sample stock solution: Equivalent to 14 mg/mL of benzocaine from Ointment in methanol Prepare by adding to a quantity of methanol corresponding to 75% of the total volume. Sonicate for about 1 min, and dilute to volume. Sample solution: 1.4 mg/mL from the Sample stock solution in Diluent Prepare by adding a quantity of the Sample stock solution corresponding to 10% of the total volume to a quantity of Diluent corresponding to 75% of the total volume. Shake, and dilute to volume with Diluent. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 313 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for benzocaine, butamben, and tetracaine are 0.3, 0.8, and 1.0, respectively.] Suitability requirements Resolution: NLT 2 between the benzocaine peak and the butamben peak and between the butamben peak and the tetracaine peak Relative standard deviation: NMT 2.0% for each of the three analyte peaks
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing. USP REFERENCE STANDARDS 11 USP Benzocaine RS USP Butamben RS USP Tetracaine Hydrochloride RS
Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Solution

(Comment on this Monograph)id=m8108=Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Solution=BMonos.pdf) DEFINITION Benzocaine, Butamben, and Tetracaine Hydrochloride Topical Solution contains NLT 90.0% and NMT 110.0% of the labeled amounts of benzocaine (C9H11NO2), butamben (C11H15NO2), and tetracaine hydrochloride (C15H24N2O2 HCl). IDENTIFICATION The retention times of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol and water (3:2) Mobile phase: Methanol, 0.25 M sodium 1heptanesulfonate, and water (25:1:25) Standard solution: Transfer 140 mg of USP Benzocaine RS, in a 100-mL volumetric flask with the aid of 25 mL of methanol, and swirl. Transfer 140 J mg of USP Butamben RS to the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, in percentage of butamben to the labeled amount, in percentage of benzocaine in the Topical Solution. Transfer 140 J mg of USP Tetracaine Hydrochloride RS, into the same volumetric flask with the aid of 25 mL of water, J being the ratio of the labeled amount, in percentage of tetracaine hydrochloride to the labeled amount, in percentage of benzocaine in the Topical Solution. Sonicate for 1 min, and dilute with Diluent to volume.
USP 32
Sample stock solution: Equivalent to 14 mg/mL of benzocaine from a volume of Topical Solution in methanol Prepare by adding to a quantity of methanol corresponding to 75% of the total volume. Sonicate for 1 min, and dilute to volume. Sample solution: 1.4 mg/mL from the Sample stock solution in Diluent Prepare by adding a quantity of the Sample stock solution corresponding to 10% of the total volume to a quantity of Diluent corresponding to 75% of the total volume, shake, and dilute to volume with Diluent. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 313 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for benzocaine, butamben, and tetracaine are 0.3, 0.8, and 1.0, respectively.] Suitability requirements Resolution: NLT 2 between the benzocaine peak and the butamben peak and between the butamben peak and the tetracaine peak Relative standard deviation: NMT 2.0% for each of the three analyte peaks Analysis Samples: Sample solution and Standard solution Calculate the individual percentages of C9H11NO2, C11H15NO2, and C15H24N2O2 HCl in the portion of Topical Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the corresponding analyte from the Sample solution = peak response from the corresponding analyte rS from the Standard solution = concentration of the appropriate Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of the analyte in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MINIMUM FILL 755: Meets the requirements rU

IDENTIFICATION A. PROCEDURE Sample solution: Transfer an amount of Topical Aerosol, equivalent to 5 mg of benzocaine, to a beaker, add 20 mL of 0.5 N hydrochloric acid, warm gently to disperse the solution, cool, and filter, if necessary, to obtain a clear solution. Analysis: To 10 mL of the clear Sample solution, add 5 drops of a solution of sodium nitrite (1 in 10), and 2 drops of methyl red TS, and neutralize with 1 N sodium hydroxide. Add 2 mL of 20 mg 2-naphthol/mL 1 N sodium hydroxide. Acceptance criteria: An orange-red precipitate is formed. B. The retention time of the major peak for benzocaine of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. The retention time of the major peak for menthol of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY BENZOCAINE Diluent: Methanol and water (1:1) Mobile phase: Methanol, glacial acetic acid, and water (14:1:10) Standard solution: 0.02 mg/mL of USP Benzocaine RS in Diluent Sample stock solution: Spray the Topical Aerosol into a flask, and dilute a volume of this solution with Diluent to prepare a nominal 2 mg/mL solution. Sample solution: Equivalent to 0.02 mg/mL from the Sample stock solution, diluted with Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 294 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H11NO2 in the portion of Topical Aerosol taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Benzocaine RS in the Standard solution (mg/mL) CU = nominal concentration in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% MENTHOL Internal standard solution: 1 mg/mL of decanol in nhexane Standard stock solution: 1 mg/mL of USP Menthol RS in n-hexane Standard solution: Transfer 5.0 mL of Standard stock solution and 5.0 mL of Internal standard solution to a 100-mL volumetric flask, and dilute with ether to volume. Sample stock solution: Spray the Topical Aerosol into a flask. Transfer an amount nominally equivalent to 50 mg of menthol to a separator, and extract with three 15-mL portions of ether, collecting the ether extracts in a 50-mL volumetric flask. Dilute with ether to volume. rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing. USP REFERENCE STANDARDS 11 USP Benzocaine RS USP Butamben RS USP Tetracaine Hydrochloride RS
Benzocaine and Menthol Topical Aerosol

(Comment on this Monograph)id=m8102=Benzocaine and Menthol Topical Aerosol=B-Monos.pdf) DEFINITION Benzocaine and Menthol Topical Aerosol is a solution of Benzocaine and Menthol with suitable propellants in a pressurized container. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of benzocaine (C9H11NO2) and menthol (C10H20O).
40
USP 32
Sample solution: Transfer 5.0 mL of the Sample stock solution to a 100-mL volumetric flask, add 5.0 mL of nhexane and 5.0 mL of Internal standard solution, and dilute with ether to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m column packed with 10% phase G16 on support S1AB Carrier gas: Dry helium Temperature Column: 170 Injection port: 260 Detector: 240 Flow rate: 50 mL/min Injection size: 2 L System suitability Sample: Standard solution [NOTEThe relative retention times for menthol and decanol are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between the menthol and decanol peaks Relative standard deviation: NMT 2.0% of the ratio of the peak response of menthol to that of decanol Analysis Samples: Standard solution and Sample solution Calculate the percentage C10H20O in each mL of Topical Aerosol taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of menthol to decanol from the Sample solution RS = peak response ratio of menthol to decanol from the Standard solution CS = concentration of USP Menthol RS in the Standard solution (mg/mL) CU = nominal concentration of menthol in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements for absence of Staphylococcus aureus and Pseudomonas aeruginosa. OTHER REQUIREMENTS: It meets the requirements under Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers 601, Pressure Test and Leakage Test. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, pressurized containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Benzocaine RS USP Menthol RS RU
Benzoic Acid
(Comment on this Monograph)id=m8130=Benzoic Acid=BMonos.pdf)
C7H6O2 Benzoic acid; Benzoic acid [65-85-0].
122.12
DEFINITION Benzoic Acid contains NLT 99.5% and NMT 100.5% of C7H6O2, calculated on the anhydrous basis. IDENTIFICATION PROCEDURE Sample solution: Prepare a saturated solution of benzoic acid in water, and filter twice. Analysis 1: To one portion of the filtrate, add ferric chloride TS. Acceptance criteria 1: A salmon-colored precipitate is formed. Analysis 2: To a separate 10-mL portion of the filtrate, add 1 mL of 7 N sulfuric acid and cool the mixture. Acceptance criteria 2: A white precipitate forms in 10 min; this precipitate is soluble in ether. ASSAY PROCEDURE Sample: 500 mg of benzoic acid Analysis: Dissolve in 25 mL of diluted alcohol that previously has been neutralized with 0.1 N sodium hydroxide, add phenolphthalein TS, and titrate with 0.1 N sodium hydroxide VS to a pink color. Each mL of 0.1 N sodium hydroxide is equivalent to 12.21 mg of C7H6O2. Acceptance criteria: 99.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.05% HEAVY METALS 231: NMT 10 ppm Sample solution: Dissolve 2.0 g in 25 mL of acetone, and add 2 mL of water. Analysis: Add 1.2 mL of thioacetamide-glycerin base TS and 2 mL of pH 3.5 Acetate Buffer, and allow to stand for 5 min. Acceptance criteria: Any color produced is not darker than that of a control made with 25 mL of acetone, 2.0 mL of Standard Lead Solution, and treated in the same manner. SPECIFIC TESTS CONGEALING TEMPERATURE 651: 121123 WATER DETERMINATION, Method I 921: NMT 0.7%, a 1 in 2 solution of methanol in pyridine being used as the solvent
USP 32
READILY CARBONIZABLE SUBSTANCES TEST 271: 500 mg in 5 mL of sulfuric acid TS: the solution has no more color than Matching Fluid Q. READILY OXIDIZABLE SUBSTANCES: Add 1.5 mL of sulfuric acid to 100 mL of water, heat to boiling, and add 0.1 N potassium permanganate, dropwise, until the pink color persists for 30 s. Dissolve 1.00 g of benzoic acid in the hot solution, and titrate with 0.1 N potassium permanganate VS to a pink color that persists for 15 s: NMT 0.50 mL of 0.10 N potassium permanganate is consumed. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Official Monographs / Benzoic 41

Similarly, mix 4 g of chromatographic siliceous earth with 3 mL of Ferric chloride-urea reagent, and pack uniformly over the first layer. Cover the column with a pad of glass wool. Column B: Insert a small pledget of glass wool above the stem constriction of a second 20- 2.5-cm chromatographic tube. Mix 4 g of chromatographic siliceous earth with 2 mL of sodium bicarbonate solution (1 in 12), prepared just prior to use, to a uniform, fluffy mixture, and pack evenly over the glass wool. Cover the column with a pad of glass wool. Diluent: Mixture of chloroform and glacial acetic acid (97:3) Standard solution A: 20 g/mL of USP Salicylic Acid RS in Diluent Standard solution B: 40 g/mL of USP Benzoic acid RS in Diluent Sample solution: Transfer an amount of the Ointment equivalent to 100 mg of benzoic acid and 50 mg of salicylic acid to a 250-mL volumetric flask, and dissolve in 150 mL of chloroform by warming on a steam bath. Cool, dilute with chloroform to volume to obtain a solution having a nominal concentration of 200 g/mL of salicylic acid and 400 g/mL of benzoic acid. Analysis Samples: Standard solution A, Standard solution B, and Sample solution Mount Column A directly over Column B, then pipet 10 mL of Sample solution onto Column A, and allow it to pass into the column. Wash the columns with two 40-mL portions of chloroform, allowing the first portion to recede to the top of each column before adding the second portion. Discard the eluates, and separate the columns. Salicylic acid content: Elute Column A with 95 mL of Diluent collecting the eluate in a 100-mL volumetric flask. Dilute the contents of the flask with Diluent to volume, and mix. Concomitantly determine the absorbances of the eluate and Standard solution A in 1-cm cells at the wavelength of maximum absorbance at 311 nm, with a suitable spectrophotometer, using the Diluent as the blank. Calculate the percentage of C7H6O3 in the portion of Ointment: Result = (AU/AS) (CS/CU) F 100 = absorbance of the diluted eluate from Column A = absorbance of Standard solution A = concentration of USP Salicylic Acid RS in Standard solution A (g/mL) = nominal concentration of the salicylic acid in the CU Sample solution (g/mL) F = sample dilution factor, 10 Acceptance criteria: 90.0%110.0% Benzoic acid content: Elute Column B with 95 mL of a solution (3 in 100) of glacial acetic acid in chloroform, collecting the eluate in a 100-mL volumetric flask. Dilute the contents of the flask with the same solvent to volume, and mix. Concomitantly determine the absorbances of eluate and Standard solution B in 1-cm cells at the wavelength of maximum absorbance at 275 nm, with a suitable spectrophotometer, using the Diluent as the blank. Calculate the percentage of C7H6O2 in the portion of Ointment taken: AU AS CS Result = (AU/AS) (CS/CU) F 100 AU AS CS CU F = absorbance of the diluted eluate from Column B = absorbance of Standard solution B = concentration of USP Benzoic Acid RS in Standard solution B (g/mL) = nominal concentration of benzoic acid in the Sample solution (g/mL) = sample dilution factor,10
Benzoic and Salicylic Acids Ointment

(Comment on this Monograph)id=m8160=Benzoic and Salicylic Acids Ointment=B-Monos.pdf) DEFINITION Benzoic and Salicylic Acids Ointment is Benzoic Acid and Salicylic Acid, present in a ratio of 2 to 1, in a suitable ointment base. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of benzoic acid (C7H6O2) and salicylic acid (C7H6O3). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Diluent: Mixture of chloroform and methanol (1:1) Standard solution A: 2.4 mg/mL of USP Benzoic Acid RS in Diluent Standard solution B: 1.2 mg/mL of USP Salicylic Acid RS in Diluent Sample solution: Equivalent to 60 mg of benzoic acid and 30 mg of salicylic acid from Ointment, in 25 mL of Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L of each solution at separate points 2.5 cm from the bottom edge of a 20- 20-cm thin-layer chromatographic plate Developing solvent system: Chloroform, acetone, isopropyl alcohol, methanol, and ammonium hydroxide (30:30:15:15:10) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chromatographic chamber, mark the solvent front, and allow the solvent to evaporate. View the chromatogram under short-wavelength (254 nm) UV radiation. Acceptance criteria: The two major fluorescent spots from the Sample solution correspond in color and in RF value to those from the respective Standard solution A and B. ASSAY PROCEDURE Ferric chloride-urea reagent: On the day of use, dissolve, without heating, 18 g of urea in a mixture of 2.5 mL of ferric chloride solution (6 in 10) and 12.5 mL of 0.05 N hydrochloric acid. Column A: Insert a small pledget of glass wool above the stem constriction of a 20- 2.5-cm chromatographic tube. Mix 1 g of chromatographic siliceous earth with 0.5 mL of dilute phosphoric acid (3 in 10) to form a uniform, fluffy mixture, transfer to the chromatographic tube, and pack evenly over the glass wool, exerting gentle pressure.
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USP 32
the Benzoin taken for the Assay, and subtract it from the original weight of the Benzoin taken. The difference between this result and the weight of the residue in the extraction thimble represents the alcohol-soluble extractive. Acceptance criteria: The alcohol-soluble extractive is NLT 75.0% in Sumatra Benzoin and NLT 90.0% in Siam Benzoin. OTHER COMPONENTS CONTENT OF BENZOIC ACID Sample solution: 1 g of powdered Benzoin with 15 mL of warm carbon disulfide Filter through a small pledget of cotton, wash the cotton with an additional 5 mL of carbon disulfide, and allow the filtrate to evaporate spontaneously. Acceptance criteria: Compared to the weight of Benzoin taken, the weight of the residue is NLT 6.0% in Sumatra Benzoin and NLT 12.0% in Siam Benzoin. This residue meets the requirements for Identification TestsGeneral 191, Benzoate. SPECIFIC TESTS ARTICLES OF BOTANICAL ORIGIN, Acid-Insoluble Ash 561: NMT 1.0% in Sumatra Benzoin; NMT 0.5% in Siam Benzoin ARTICLES OF BOTANICAL ORIGIN, Foreign Organic Matter 561: NMT 1.0% in Siam Benzoin ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate whether it is Sumatra Benzoin or Siam Benzoin.
PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and avoid exposure to temperatures exceeding 30. LABELING: Label Ointment to indicate the concentrations of Benzoic Acid and Salicylic Acid and to indicate whether the ointment base is water-soluble or water-insoluble. USP REFERENCE STANDARDS 11 USP Benzoic Acid RS USP Salicylic Acid RS
Benzoin
(Comment on this Monograph)id=m8190=Benzoin=BMonos.pdf) DEFINITION Benzoin is the balsamic resin obtained from Styrax benzoin Dryander or Styrax paralleloneurus Perkins, known in commerce as Sumatra Benzoin, or from Styrax tonkinensis (Pi rre) Craib ex e Hartwich, or other species of the Section Anthostyrax of the genus Styrax, known in commerce as Siam Benzoin (Fam. Styraceae). Sumatra Benzoin yields NLT 75.0% of alcohol-soluble extractive, and Siam Benzoin yields NLT 90.0% of alcohol-soluble extractive. Botanic characteristics Sumatra Benzoin: Blocks or lumps of varying size, made up of tears, compacted together, with a reddish brown, reddish gray, or grayish brown resinous mass; the tears are externally yellowish or rusty brown, milky white on fresh fracture; hard and brittle at ordinary temperatures, but softened by heat and becoming gritty on chewing. Siam Benzoin: Pebble-like tears of variable size and shape, compressed, yellowish brown to rusty brown externally, milky white on fracture, separate or very slightly agglutinated; hard and brittle at ordinary temperatures, but softened by heat and becoming plastic on chewing. IDENTIFICATION A. A solution in alcohol becomes milky upon the addition of water, and the mixture is acid to litmus paper. B. Heat a few fragments in a test tube. Sumatra Benzoin evolves a sublimate consisting of plates and small, rod-like crystals of cinnamic acid and its esters that strongly polarize light. Siam Benzoin evolves a sublimate directly above the melted mass, consisting of numerous long, rod-shaped crystals of benzoic acid that do not strongly polarize light. C. Heat 500 mg in a test tube with 10 mL of potassium permanganate TS: only the Sumatra variety develops a faint odor of benzaldehyde. ASSAY PROCEDURE Sample: Place 2 g of Benzoin in a tared extraction thimble, and insert the thimble in a continuous-extraction apparatus. Place 100 mg of sodium hydroxide in the receiving flask of the apparatus, and extract the Benzoin with alcohol for 5 h, or until completely extracted. Dry the extraction thimble containing the insoluble residue at 105 for 2 h. Analysis: On a separate portion of Benzoin, determine the water content as directed for Water Determination 921, Method II. Calculate the weight of water in the quantity of
Compound Benzoin Tincture

(Comment on this Monograph)id=m8220=Compound Benzoin Tincture=B-Monos.pdf) DEFINITION Prepare Compound Benzoin Tincture as follows.
Benzoin, in moderately coarse powder Aloe, in moderately coarse powder Storax Tolu Balsam To make 100 g 20 g 80 g 40 g 1000 mL
Prepare a Tincture by Process M (see Pharmaceutical Dosage Forms 1151), using alcohol as the menstruum. OTHER COMPONENTS Alcohol Determination, Method II 611: 74.0%80.0% of C2H5OH, the dilution to approximately 2% alcohol being made with methanol instead of with water SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.8700.885 LIMIT OF NONVOLATILE RESIDUE: Evaporate 3 mL of Tincture in a suitable tared dish on a steam bath, and dry the residue at 100 for 2 h. Acceptance criteria: The weight of the residue is NLT 525 mg and NMT 675 mg. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and avoid exposure to direct sunlight and to excessive heat.
USP 32
LABELING: Label it to indicate that it is flammable.
Official Monographs / Benzonatate 43
Benzonatate Capsules
(Comment on this Monograph)id=m8260=Benzonatate Capsules=B-Monos.pdf) DEFINITION Benzonatate Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of benzonatate (C30H53NO11 av.). IDENTIFICATION A. INFRARED ABSORPTION 197F [NOTEIf a difference is observed, or if excipients are present, use the equivalent of 100 mg of benzonatate from Capsules. Mix with 25 mL of 0.01 N hydrochloric acid, and proceed as directed under IdentificationOrganic Nitrogenous Bases 181, beginning with Transfer the liquid to a separator.] B. ULTRAVIOLET ABSORPTION 197U Solution: 15 g/mL from Capsules ASSAY PROCEDURE Standard solution: 0.5 mg/mL of USP Benzonatate RS Sample stock solution: Mix a number of capsules, equivalent to 500 mg of benzonatate, with 40 mL of chloroform in a suitable high speed blender, and dilute with chloroform to 100 mL. Sample solution: 10.0 mL of Sample stock solution, in a 100-mL volumetric flask, evaporate the chloroform on a steam bath with the aid of a current of air. Dissolve the residue in water and dilute to volume. Blank: Water Spectrometric conditions Mode: UV-Vis Analytical wavelength: 500 nm Cell: 1 cm Analysis Samples: Standard solution, Sample solution, and Blank Transfer 4.0 mL each of the Standard solution, the Sample solution, and Blank, to separate test tubes. To each tube add in succession 1.0 mL of 1 M hydroxylamine hydrochloride and 1.0 mL of 3.5 N sodium hydroxide, mixing after each addition. Allow to stand for 10 min, accurately timed, then add 1.0 mL of 3.5 N hydrochloric acid, mix, add 1.0 mL of an 80 mg/mL ferric chloride solution, and mix. Allow to stand for 30 min, accurately timed. Gently swirl the tubes for 1 min to remove any gas bubbles present, then concomitantly determine the absorbances of the solution Calculate the percentage of C30H53NO11 av in the number of Capsules: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Benzonatate RS in the Standard solution (g/mL) = nominal concentration of benzonatate in the CU Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of benzonatate dissolved by employing the following method. Mobile phase: Acetonitrile and 0.04 M monobasic potassium phosphate (3:1) AU AS CS
Benzonatate
(Comment on this Monograph)id=m8250=Benzonatate=BMonos.pdf)
C30H53NO11 (av.) 603.00 (av.) Benzoic acid, 4-(butylamino)-, 2,5,8,11,14,17,20,23,26nonaoxaoctacos-28-yl ester; 2,5,8,11,14,17,20,23,26-Nonaoxaoctacosan-28-yl p(butylamino)benzoate [104-31-4]. DEFINITION Benzonatate contains NLT 95.0% and NMT 105.0% of C30H53NO11. IDENTIFICATION A. INFRARED ABSORPTION 197F B. ULTRAVIOLET ABSORPTION 197U Solution: 15 g/mL ASSAY PROCEDURE Sample: 5 g of Benzonatate Analysis: Reflux with 25.0 mL of 0.5 N sodium hydroxide VS for 1 h. Cool, add 25 mL of water and 10 drops of bromothymol blue TS, and titrate the excess alkali with 0.5 N hydrochloric acid VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.5 N sodium hydroxide is equivalent to 301.5 mg of C30H53NO11. Acceptance criteria: 95.0%105.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221: Mix 20 mL of a solution (1 in 10) with 20 mL of water and 1 mL of nitric acid, shake for 1 h, and allow to stand for 1 h. Pass through a filter having a porosity of 0.2 m, and to the filtrate add 1 mL of silver nitrate TS. Dilute with water to 50 mL, mix, and allow to stand protected from light for 10 min. Acceptance criteria: the turbidity does not exceed that produced by 0.10 mL of 0.020 N hydrochloric acid (35 ppm). CHLORIDE AND SULFATE, Sulfate 221: Mix 5 mL of a solution (1 in 20) with 5 mL of water and 1 mL of 3 N hydrochloric acid, shake for 1 h, and allow to stand for 1 h. Pass through a filter having a porosity of 0.2 m, and to the filtrate add 1 mL of barium chloride TS. Mix, and allow to stand for 10 min. Acceptance criteria: the turbidity does not exceed that produced by 0.10 mL of 0.020 N sulfuric acid (400 ppm). HEAVY METALS, Method II 231: NMT 10 ppm SPECIFIC TESTS REFRACTIVE INDEX 831: 1.5091.511 at 20 WATER DETERMINATION, Method I 921: NMT 0.3% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Benzonatate RS
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USP 32
Sample solution: 10 mg/mL of benzoyl peroxide in methanol Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Toluene, dichloromethane, and glacial acetic acid (50:2:1) Analysis Samples: Standard solution and Sample solution Analysis: Place the plate in a developing chamber containing, and equilibrated with the Developing solvent system. Develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate, and allow the solvent to evaporate. Observe the plate under short-wavelength UV light. Aceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. The Sample solution in the Organic Impurities test exhibits a major peak for benzoyl peroxide, the retention time of which corresponds to that exhibited by the Standard solution. ASSAY PROCEDURE Sample: 300 mg of previously mixed Hydrous Benzoyl Peroxide in a conical flask fitted with a ground-glass stopper, and weigh again to obtain the weight of the Sample. Analysis: Add 30 mL of glacial acetic acid, previously sparged with carbon dioxide for not less than 2 min just before use, and swirl the flask gently to effect solution. Add 5 mL of potassium iodide solution (1 in 2), and mix. Allow the solution to stand for 1 min. Titrate the liberated iodine with 0.1 N sodium thiosulfate VS. As the endpoint is approached, add 1 drop of starch iodide paste TS, or equivalent, and continue the titration to the discharge of the blue color. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N sodium thiosulfate is equivalent to 12.11 mg of C14H10O4. Acceptance criteria: 65.0%82.0% IMPURITIES Organic Impurities PROCEDURE Solution A: Prepare a mixture of acetonitrile and glacial acetic acid (1000:1). Solution B: Prepare a mixture of water and glacial acetic acid (1000:1). Mobile phase: Use variable mixtures of Solution A and Solution B as directed for Chromatographic system. Sample solution: 0.32 mg/mL of benzoyl peroxide in acetonitrile Standard solution: Dissolve a quantity of Hydrous Benzoyl Peroxide, previously subjected to the Assay, in acetonitrile to obtain a solution containing 0.32 mg/mL. System suitability solution: 100 g/mL of benzoic acid and 60 g/mL of methylparaben in acetonitrile Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L The chromatograph is programmed as follows.
Standard solution: 0.1 mg/mL of USP Benzonatate RS [NOTESonicate as necessary.] Sample solutions: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Pass a portion of the solution under test through a 0.45-m filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 310 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 15 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of benzonatate dissolved: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benzonatate RS in the Standard solution (mg/mL) = nominal concentration of benzonatate in the CU Sample solution (mg/mL) Tolerances: NLT 80% (Q) of the labeled amount of benzonatate UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Benzonatate RS
Hydrous Benzoyl Peroxide

(Comment on this Monograph)id=m8310=Hydrous Benzoyl Peroxide=B-Monos.pdf)
C14H10O4 (anhydrous) Peroxide, dibenzoyl; Benzoyl peroxide [94-36-0].
242.23
DEFINITION Hydrous Benzoyl Peroxide contains NLT 65.0% and NMT 82.0% of C14H10O4. It contains 26% of water for the purpose of reducing flammability and shock sensitivity. [CAUTIONHydrous Benzoyl Peroxide may explode at temperatures higher than 60 or cause fires in the presence of reducing substances. Store it in the original container, treated to reduce static charges.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 10 mg/mL of Hydrous Benzoyl Peroxide, previously subjected to the Assay, in methanol
USP 32
Time (min) 0 20 30 Solution A (%) 18 60 60 Solution B (%) 82 40 40
Official Monographs / Benzoyl 45

Sample stock solution: Equivalent to 40 mg of benzoyl peroxide from Gel. In a 50-mL volumetric flask, add 40 mL of acetonitrile, and shake until the material is thoroughly dispersed. Sonicate the mixture for 5 min, dilute with acetonitrile to volume, mix, and filter. Sample solution: 10 mL of Sample stock solution and 5 mL of Internal standard solution. Dilute with acetonitrile to 25 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution (three replicate injections) [NOTEThe lowest and highest peak response ratios (RS) agree within 2.0%. The retention times for ethyl benzoate and benzoyl peroxide are 7 and 14 min, respectively.] Suitability requirements Resolution: NLT 2.0 between ethyl benzoate and benzoyl peroxide Tailing factors: NMT 2.0 for the ethyl benzoate and benzoyl peroxide peaks Analysis Samples: Sample solution and Standard solution Calculate the percentage of C14H10O4 in the portion of Gel taken: Result = (RU/RS) (CS/CU) 100 = peak response ratios of benzoyl peroxide to ethyl benzoate from the Sample solution RS = peak response ratios of benzoyl peroxide to ethyl benzoate from the Standard solution = concentration of benzoyl peroxide in the CS Standard solution (mg/mL) = nominal concentration of benzoyl peroxide in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%125.0% IMPURITIES Organic Impurities PROCEDURE Solution A: Acetonitrile and glacial acetic acid (1000:1) Solution B: Glacial acetic acid and water (1:1000) Mobile phase: See the gradient table below.
System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 2.0 between benzoic acid and methylparaben Tailing factors: NMT 2.0 for the benzoic acid and methylparaben peaks Analysis Samples: Sample solution and Standard solution Calculate the area, as a percentage, of each peak in the chromatogram of the Sample solution: Result = (rU/rT) 100 = peak response for any individual peak other than the principal peak in the Sample solution = sum of the peak responses of all the individual rT peaks including the principal peak in the Sample solution Acceptance criteria: The area of any individual peak other than the principal peak is NMT 1.5% of the total area. The sum of the areas of all peaks other than the principal peak is NMT 2.0% of the total area. rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Store in the original container, at room temperature. [NOTEDo not transfer Hydrous Benzoyl Peroxide to metal or glass containers fitted with friction tops. Do not return unused material to its original container, but destroy it by treatment with sodium hydroxide solution (1 in 10) until addition of a crystal of potassium iodide results in no release of free iodine.]
RU
Benzoyl Peroxide Gel

(Comment on this Monograph)id=m8320=Benzoyl Peroxide Gel=B-Monos.pdf) DEFINITION Benzoyl Peroxide Gel is benzoyl peroxide in a suitable gel base. It contains NLT 90.0% and NMT 125.0% of the labeled amount of benzoyl peroxide (C14H10O4). IDENTIFICATION The retention time of the major peak for benzoyl peroxide of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile in water (5 in 10) Internal standard solution: 3.6 mg/mL of ethyl benzoate in acetonitrile Standard stock solution: A suitable quantity of benzoyl peroxide, recently subjected to the Assay under Hydrous Benzoyl Peroxide, in a weighed conical flask fitted with a glass stopper. Weigh again to obtain the weight of the specimen, and quantitatively dissolve in acetonitrile to obtain a solution containing 0.8 mg/mL. Standard solution: 10 mL of Standard stock solution and 5 mL of Internal standard solution Dilute with acetonitrile to 25 mL. [NOTEThis Standard solution contains 0.32 mg/mL of benzoyl peroxide.]
System suitability solution: 100 g/mL of benzoic acid and 60 g/mL of methylparaben in acetonitrile Standard solution A: 500 g/mL of benzoic acid in acetonitrile Standard solution B: 20 g/mL of ethyl benzoate in acetonitrile Standard solution C: 20 g/mL of benzaldehyde in acetonitrile Standard solution D: Equivalent to 40 g/mL of anhydrous benzoyl peroxide, previously subjected to the Assay under Hydrous Benzoyl Peroxide, in acetonitrile Sample solution: Equivalent to 100 mg of benzoyl peroxide from Gel. To a 50-mL volumetric flask, add 25 mL of acetonitrile, shake vigorously to disperse the specimen,
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sonicate for 5 min, dilute with acetonitrile to volume, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 2.0 between benzoic acid and methylparaben Tailing factors: NMT 2.0 for the benzoic acid and methylparaben peaks Analysis Samples: Sample solution and Standard solution Acceptance criteria: The responses of any peaks from the Sample solution corresponding to benzoic acid, ethyl benzoate, and benzaldehyde are NMT those of the main peaks from Standard solution A (25%), Standard solution B (1%), and Standard solution C (1%), respectively. The response of any other impurity peak from the Sample solution, other than the main benzoyl peroxide peak, any benzoic acid, ethyl benzoate, benzaldehyde, methylparaben, or propylparaben peak, and any solvent peak, is NMT that from Standard solution D (2%); and the sum of the responses of all the impurity peaks, other than those of benzoic acid, ethyl benzoate, and benzaldehyde, is NMT that from Standard solution D (2%).
USP 32
Acceptance criteria: The RF value of the principal spot from the solution under test corresponds to that from the Standard solution. B. The retention time of the major peak for benzoyl peroxide of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile in water (5 in 10) Internal standard solution: 3.6 mg/mL of ethyl benzoate in acetonitrile Standard stock solution: A suitable quantity of benzoyl peroxide, recently subjected to the Assay under Hydrous Benzoyl Peroxide, in a weighed conical flask fitted with a glass stopper. Weigh again to obtain the weight of the specimen, and quantitatively dissolve in acetonitrile to obtain a solution containing 0.8 mg/mL. Standard solution: 10 mL of Standard stock solution and 5 mL of Internal standard solution. Dilute with acetonitrile to 25 mL. [NOTEThis Standard solution contains 0.32 mg/mL of benzoyl peroxide.] Sample stock solution: Equivalent to 40 mg of benzoyl peroxide from Lotion. In a 50-mL volumetric flask, add 40 mL of acetonitrile. Shake vigorously to disperse the specimen, and sonicate until the material is thoroughly dispersed. Sonicate the mixture for 5 min, dilute with acetonitrile to volume, mix, and filter. Sample solution: 10 mL of Sample stock solution and 5 mL of Internal standard solution. Dilute with acetonitrile to 25 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution (three replicate injections) [NOTEThe lowest and highest peak response ratios (RS) agree within 2.0%. The retention times for ethyl benzoate and benzoyl peroxide are 7 and 14 min, respectively.] Suitability requirements Resolution: NLT 2.0 between ethyl benzoate and benzoyl peroxide Tailing factors: NMT 2.0 for the ethyl benzoate and benzoyl peroxide peaks Analysis Samples: Sample solution and Standard solution Calculate the percentage of C14H10O4 in the portion of Lotion taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = peak response ratios of benzoyl peroxide to ethyl benzoate from the Sample solution = peak response ratios of benzoyl peroxide to ethyl benzoate from the Standard solution = concentration of benzoyl peroxide in the Standard solution (mg/mL) = nominal concentration of benzoyl peroxide in the Sample solution (mg/mL)
SPECIFIC TESTS PH 791: 2.86.6 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Benzoyl Peroxide Lotion

(Comment on this Monograph)id=m8330=Benzoyl Peroxide Lotion=B-Monos.pdf) DEFINITION Benzoyl Peroxide Lotion is benzoyl peroxide in a suitable lotion base. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C14H10O4. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 10 mg/mL of Hydrous Benzoyl Peroxide, previously subjected to the Assay, in methanol Sample solution: Equivalent to 10 mg/mL of benzoyl peroxide from Lotion in acetone Application volume: 5 L Developing solvent system: Toluene, dichloromethane, and glacial acetic acid (50:2:1) Analysis Sample: Sample solution and Standard solution Place the plate in a developing chamber containing, and equilibrated, with Developing solvent system. Develop the chromatogram until the solvent front has moved threefourths of the length of the plate. Remove the plate, and allow the solvent to evaporate. Observe the plate under short-wavelength UV light.
USP 32
Official Monographs / Benztropine 47

SPECIFIC TESTS PH 791: 2.86.6 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
IMPURITIES Organic Impurities PROCEDURE Solution A: Acetonitrile and glacial acetic acid (1000:1) Solution B: Glacial acetic acid and water (1:1000) Mobile phase: See the gradient table below.
Benztropine Mesylate
(Comment on this Monograph)id=m8500=Benztropine Mesylate=B-Monos.pdf)
System suitability solution: 100 g/mL of benzoic acid and 60 g/mL of methylparaben in acetonitrile Standard solution A: 500 g/mL of benzoic acid in acetonitrile Standard solution B: 20 g/mL of ethyl benzoate in acetonitrile Standard solution C: 20 g/mL of benzaldehyde in acetonitrile Standard solution D: Equivalent to 40 g/mL of anhydrous benzoyl peroxide, previously subjected to the Assay under Hydrous Benzoyl Peroxide, in acetonitrile Sample solution: Equivalent to 100 mg of benzoyl peroxide from Lotion. In a 50-mL volumetric flask, add 25 mL of acetonitrile, shake vigorously to disperse the specimen, sonicate for 5 min, dilute with acetonitrile to volume, mix, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 2.0 between benzoic acid and methylparaben Tailing factors: NMT 2.0 for the benzoic acid and methylparaben peaks Analysis Samples: Sample solution and Standard solution Acceptance criteria: The responses of any peaks from the Sample solution corresponding to benzoic acid, ethyl benzoate, and benzaldehyde are NMT those of the main peaks from Standard solution A (25%), Standard solution B (1%), and Standard solution C (1%), respectively. The response of any other impurity peak from the Sample solution, other than the main benzoyl peroxide peak, any benzoic acid, ethyl benzoate, benzaldehyde, methylparaben, or propylparaben peak, and any solvent peak, is NMT that from Standard solution D (2%); and the sum of the responses of all the impurity peaks, other than those of benzoic acid, ethyl benzoate, and benzaldehyde, is NMT that from Standard solution D (2%).
403.54 C21H25NO CH4O3S 8-Azabicyclo[3.2.1]octane, 3-(diphenylmethoxy)-, endo-, methanesulfonate; 3-(Diphenylmethoxy)-1H,5H-tropane methanesulfonate [132-17-2]. DEFINITION Benztropine Mesylate contains NLT 98.0% and NMT 100.5% of C21H25NO CH4O3S, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 60 mg of Benztropine Mesylate Analysis: Dissolve in 25 mL of water, add 5 mL of sodium carbonate TS, and extract with four 10-mL portions of chloroform. Wash the combined chloroform extracts with about 10 mL of water, and extract the wash solution with 5 mL of chloroform. Filter the combined chloroform extracts through a tightly packed pledget of cotton, and wash the cotton with about 5 mL of chloroform. Add methyl red TS, and titrate the chloroform solution with 0.01 N perchloric acid in dioxane VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.01 N perchloric acid is equivalent to 4.035 mg of C21H25NO CH4O3S. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
NMT 0.1%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 141148 LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Benztropine Mesylate RS
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USP 32
Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C21H25NO CH4O3S in each mL of the Injection: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benztropine Mesylate RS in the Standard solution (mg/mL) = nominal concentration of benztropine mesylate CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 55.6 USP Endotoxin Units/mg of benztropine mesylate PH 791: 5.08.0 OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Benztropine Mesylate RS USP Endotoxin RS
Benztropine Mesylate Injection

(Comment on this Monograph)id=m8530=Benztropine Mesylate Injection=B-Monos.pdf) DEFINITION Benztropine Mesylate Injection is a sterile solution of Benztropine Mesylate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C21H25NO CH4O3S. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard stock solution: 10 mg of USP Benztropine Mesylate RS in 50 mL in a separator (0.2 mg/mL) Standard solution: To the separator containing the Standard stock solution, add 2 mL of 1 N sodium hydroxide. Extract with three 10-mL portions of chloroform, collecting the chloroform extracts to a 50-mL beaker. Evaporate the chloroform extracts with the aid of gentle heat and a current of air to dryness, and dissolve the residue in 1 mL of chloroform. Sample stock solution: Equivalent to 10 mg of Benztropine Mesylate from a volume of Injection to 50 mL in a separator (0.2 mg/mL) Sample solution: Repeat the procedure for the Standard solution, using the Sample stock solution. Application volume: 1 L Developing solvent system: Chloroform, methanol, and a 1-in-4 solution of ammonium hydroxide (40:10:1) Analysis Samples: Sample solution and Standard solution Allow the applications to dry, and develop the chromatogram in the Developing solvent system, until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by lightly spraying with potassium iodoplatinate TS. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Solution A: 0.005 M Octylamine phosphate buffer Transfer 0.83 mL of octylamine to a 1-L volumetric flask, dilute to volume, adjust with phosphoric acid to a pH of 3.0, and pass the solution through a membrane filter. Mobile phase: Acetonitrile and Solution A (13:7) Standard solution: 1 mg/mL of USP Benztropine Mesylate RS Sample solution: Equivalent to 1 mg/mL of benztropine mesylate from the volume of Injection Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 259 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min [NOTEAdjust the flow rate to obtain a retention time of 7 min for benztropine mesylate.]
Benztropine Mesylate Tablets

(Comment on this Monograph)id=m8540=Benztropine Mesylate Tablets=B-Monos.pdf) DEFINITION Benztropine Mesylate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C21H25NO CH4O3S. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard stock solution: 10 mg of USP Benztropine Mesylate RS in 50 mL in a separator (0.2 mg/mL) Standard solution: To the separator containing the Standard stock solution, add 2 mL of 1 N sodium hydroxide. Extract with three 10-mL portions of chloroform, collecting the chloroform extracts to a 50-mL beaker. Evaporate the chloroform extracts with the aid of gentle heat and a current of air to dryness, and dissolve the residue in 1 mL of chloroform. Sample stock solution: Add equivalent to 10 mg of Benztropine Mesylate from a portion of finely powdered Tablets in 50 mL, shake by mechanical means for 30 min, and filter into a separator (0.2 mg/mL). Sample solution: Repeat the procedure for the Standard solution, using the Sample stock solution.
USP 32
Application volume: 1 L Developing solvent system: Chloroform, methanol, and a 1-in-4 solution of ammonium hydroxide (40:10:1) Analysis Samples: Sample solution and Standard solution Allow the applications to dry, and develop the chromatogram in the Developing solvent system, until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by lightly spraying with potassium iodoplatinate TS. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Solution A: Transfer 0.83 mL of octylamine to a 1-L volumetric flask, dilute to volume, adjust with phosphoric acid to a pH of 3.0, and pass the solution through a membrane filter. Diluent: Isopropyl alcohol, phosphoric acid, and water (40:0.1:60) Mobile phase: Acetonitrile and Solution A (9:11) Standard solution: 0.04 mg/mL of USP Benztropine Mesylate RS in Diluent Sample solution: Equivalent to 0.04 mg/mL of benztropine mesylate from powdered Tablets (NLT 20 Tablets) in Diluent [NOTEAdd powdered tablets to a portion of Diluent corresponding to 60% of the final volume. Mix by mechanical means for NLT 60 min, and dilute with Diluent to volume. Centrifuge a portion of this mixture, and filter the supernatant layer.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 259 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 0.7 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 4.0 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C21H25NO CH4O3S in the portion of Tablets: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benztropine Mesylate RS in the Standard solution (mg/mL) CU = nominal concentration of benztropine mesylate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of C21H25NO CH4O3S dissolved by using the following method.
Official Monographs / Benzyl 49

Solution A: Proceed as directed in the Assay. Mobile phase: Acetonitrile and Solution A (13:7) Sample solution: Sample per Dissolution 711. Use a filtered portion of the solution under test from the dissolution vessel. Standard solution: USP Benztropine Mesylate RS in Medium. Dilute to obtain a solution having a known concentration similar to that of the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 2 mL/min Injection size: 300 L Analysis Samples: Sample solution and Standard solution Calculate the percentage of C21H25NO CH4O3S dissolved: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Benztropine Mesylate RS in the Standard solution (mg/mL) = nominal concentration of benztropine mesylate CU in the Sample solution (mg/mL) Acceptance criteria: NLT 80% (Q) of the labeled amount of C21H25NO CH4O3S UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Benztropine Mesylate RS rU rS CS
Benzyl Benzoate
(Comment on this Monograph)id=m8600=Benzyl Benzoate=BMonos.pdf)
C14H12O2 Benzoic acid, phenylmethyl ester; Benzyl benzoate [120-51-4].
212.24
DEFINITION Benzyl Benzoate contains NLT 99.0% and NMT 100.5% of C14H12O2. IDENTIFICATION INFRARED ABSORPTION 197F ASSAY PROCEDURE Sample: 2 g of Benzyl Benzoate Analysis: Transfer to a conical flask fitted with a reflux condenser, add 50.0 mL of 0.5 N alcoholic potassium hydroxide VS, and boil gently for 1 h. Cool, add phenolphthalein TS, and titrate with 0.5 N hydrochloric acid VS. Perform a blank determination (see Titrimetry 541).
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Benzyl / Official Monographs

Acceptance criteria: SPECIFIC TESTS PH 791: 8.59.2 26.0%30.0% (w/w)
USP 32
Each mL of 0.5 N alcoholic potassium hydroxide is equivalent to 106.1 mg of C14H12O2. Acceptance criteria: 99.0%100.5% SPECIFIC TESTS SPECIFIC GRAVITY 841: 1.1161.120 CONGEALING TEMPERATURE 651: NLT 18.0 Congelation may be brought about by the addition of a fragment of previously congealed Benzyl Benzoate when the temperature has reached the expected congealing temperature. REFRACTIVE INDEX 831: 1.5681.570 at 20 ALDEHYDE: Transfer 10 g to a 125-mL conical flask containing 50 mL of alcohol and 5 mL of hydroxylamine hydrochloride solution (3.5 in 100), mix, and allow to stand for 10 min. Add 1 mL of bromophenol blue TS, and titrate with 0.1 N sodium hydroxide VS to a light green endpoint. Perform a blank determination, and match the color of the endpoint with that of the titrated Sample solution. The net volume of 0.1 N sodium hydroxide consumed does not exceed 0.50 mL (0.05% as benzaldehyde). ACIDITY: Add 2 drops of phenolphthalein TS to 25 mL of alcohol, and add 0.020 N sodium hydroxide until a pink color is produced. Add 5.0 g of Benzyl Benzoate, and titrate with 0.020 N sodium hydroxide: NMT 1.5 mL of 0.020 N sodium hydroxide is required to restore the pink color. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, well-filled, lightresistant containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Benzyl Benzoate RS
Benzylpenicilloyl Polylysine Concentrate

(Comment on this Monograph)id=m8650=Benzylpenicilloyl Polylysine Concentrate=B-Monos.pdf) DEFINITION Benzylpenicilloyl Polylysine Concentrate has a molar concentration of benzylpenicilloyl moeity (C16H19N2O5S) of NLT 0.0125 M and NMT 0.020 M. It contains one or more suitable buffers. ASSAY PROCEDURE Solution A: 9 g of sodium chloride and 1.38 g of monobasic sodium phosphate in 900 mL. Adjust with 5 N sodium hydroxide or phosphoric acid to a pH of 7.6. Dilute to 1000 mL. Solution B: 0.07 mg/mL of mercuric chloride Sample solution: Concentrate and Solution A (1.0 in 500) Blank: Solution A Analysis Sample: Blank and Sample solution Transfer 3.0 mL of Sample solution to a spectrophotometric cell. Using a suitable spectrophotometer and using Blank, determine the initial absorbance at the wavelength of maximum absorbance at 282 nm. Add 0.02 mL of Solution B to the Sample solution in the spectrophotometric cell and determine the absorbance at the same wavelength after 1 and 3 min. Repeat the addition of 0.02-mL portions of Solution B until a maximum absorbance reading is obtained. Calculate the molar concentration of benzylpenicilloyl moiety in the Concentrate: Result = 500{[AM(3 + 0.02N)/3] AI}/MAB = highest absorbance observed = number of 0.02-mL portions of Solution B added to the Sample solution to obtain the maximum absorbance = initial absorbance AI = molar absorptivity of the penamaldate formed MA by the reaction of benzylpenicilloyl with mercuric chloride at pH 7.6, 22,325 B = length of the cell (cm) Acceptance criteria: 0.01250.020 M OTHER COMPONENTS BENZYLPENICILLOYL SUBSTITUTION Solution A: 19.69 g of sodium citrate dihydrate, 0.1 mL of pentachlorophenol, and 5 mL of 2,2-thiodiethanol in 900 mL of 0.2 N hydrochloric acid. Adjust with hydrochloric acid to a pH of 2.2, and dilute with water to 1000 mL. Ninhydrin reagent: 18 g of ninhydrin and 0.7 g of hydrindantin in 675 mL of dimethyl sulfoxide. Add 225 mL of 4 M lithium acetate solution previously adjusted with glacial acetic acid to a pH of 5.2. AM N
Benzyl Benzoate Lotion

(Comment on this Monograph)id=m8630=Benzyl Benzoate Lotion=B-Monos.pdf) DEFINITION Benzyl Benzoate Lotion contains NLT 26.0% and NMT 30.0% (w/w) of C14H12O2.
Benzyl Benzoate Triethanolamine Oleic Acid Purified Water To make about 250 mL 5g 20 g 750 mL 1000 mL
Mix the Triethanolamine with the Oleic Acid, and add the Benzyl Benzoate. Transfer the mixture to a suitable container of 2000-mL capacity, add 250 mL of Purified Water, and shake the mixture thoroughly. Finally add the remaining Purified Water, and again shake thoroughly. ASSAY PROCEDURE Sample: 5 g of Lotion in a conical flask Analysis: Add 25 mL of alcohol and 2 drops of phenolphthalein TS. Cool the solution to 15, and titrate quickly with 0.1 N sodium hydroxide to a slight pink color. Add 50.0 mL of 0.5 N alcoholic potassium hydroxide VS, connect the flask to a reflux condenser, and boil gently for 1 h. Cool, promptly add phenolphthalein TS and titrate with 0.5 N hydrochloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.5 N alcoholic potassium hydroxide is equivalent to 106.1 mg of C14H12O2.
USP 32
Standard solution: 91 g/mL (5 104 M) of USP L-Lysine Hydrochloride RS in Solution A Sample solution: 1.0 mL of Concentrate in a 10-mL volumetric flask, and dilute with water to volume. Transfer 1.0 mL of this solution to an ampul, add 1.5 mL of 6 N hydrochloric acid, and seal the ampul under nitrogen. Heat the ampul at 110 for 22 h. Transfer the contents of the ampul to a round-bottom, 50-mL flask, and dry by vacuum rotary evaporation. Dissolve the residue three times, using 5mL portions, evaporating to dryness after each dissolution. Dissolve the residue in 10 mL of Solution A. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: 570 nm Column: 1.75-mm 50-cm; packing of 8-m 8% crosslinked sulfonated divinylbenzene polystyrene cationexchange resin [NOTEThe column effluent is mixed continuously with flowing Ninhydrin reagent, and the flowing mixture is heated at 130 for 1.5 min in a reaction coil. The absorbance of the reaction mixture is measured continuously.] Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates Relative standard deviation: NMT 4.0% Analysis Sample: Standard solution and Sample solution [NOTEThe retention time for L-lysine is 57 min.] Calculate the molar concentration of lysine in the Concentrate: Result = (rU/rS) (C/1000 * Mr) F = peak response of the Sample solution = peak response of the Standard solution = concentration of USP L-Lysine Hydrochloride RS in the Standard solution (g/mL) = molecular weight of anhydrous lysine Mr hydrochloride, 182.65 F = dilution factor of the Sample solution (100) Calculate the percentage of benzylpenicilloyl substitution: rU rS C Result = (B/L) 100 = molar concentration of benzylpenicilloyl moiety in the Concentrate, as determined in the Assay L = molar concentration of lysine in the Concentrate Acceptance criteria: 50%70% IMPURITIES Organic Impurities PROCEDURE: LIMIT OF PENICILLENATE AND PENAMALDATE Sample solution: 1 mL of Concentrate in a 50-mL volumetric flask. Dilute with Solution A, prepared as directed in the Assay, to volume. Using a suitable spectrophotometer and using Solution A as a Blank, determine the absorbances at the wavelengths of maximum absorption at 322 nm and 282 nm. Calculate the molar concentration of penicillenate taken: Result = 50A322/MA1B A322 MA1 B = absorbance at 322 nm = molar absorptivity of the penicillenate moiety at pH 7.6, 26,600 = length of the cell (cm) AI MA B
Official Monographs / Benzylpenicilloyl 51

Acceptance criteria: NMT 0.00020 M Calculate the molar concentration of penamaldate taken: Result = 50A282/MA2B = absorbance at 282 nm = molar absorptivity of the penamaldate moiety at pH 7.6, 22,325 B = length of the cell (cm) Acceptance criteria: NMT 0.00060 M A282 MA2 SPECIFIC TESTS PH 791: 6.58.5, using the undiluted Concentrate ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label states that this article is not intended for direct administration to humans or animals. USP REFERENCE STANDARDS 11 USP L-Lysine Hydrochloride RS
Benzylpenicilloyl Polylysine Injection

(Comment on this Monograph)id=m8654=Benzylpenicilloyl Polylysine Injection=B-Monos.pdf) DEFINITION Benzylpenicilloyl Polylysine Injection has a molar concentration of benzylpenicilloyl moiety (C16N2H19O5S) of NLT 5.4 105 M and NMT 7.0 105 M. It contains one or more suitable buffers. ASSAY PROCEDURE Solution A: 9 g of sodium chloride and 1.38 g of monobasic sodium phosphate in 900 mL. Adjust with 5 N sodium hydroxide or phosphoric acid to a pH of 7.6. Dilute to 1000 mL. Solution B: 0.07 mg/mL of mercuric chloride Sample solution: Combine the contents of a sufficient number of containers to obtain NLT 3 mL of Injection. Transfer 3.0 mL of Injection to a 10-mL volumetric flask, and dilute with Solution A to volume. Blank: Solution A Analysis Samples: Blank and Sample solution Transfer 3.0 mL of Sample solution to a spectrophotometric cell. Using a suitable spectrophotometer and using the Blank, determine the initial absorbance at the wavelength of maximum absorbance at 282 nm. Add 0.02 mL of Solution B to the Sample solution in the spectrophotometric cell, and determine the absorbance at the same wavelength after 1 and 3 min. Repeat the addition of 0.02mL portions of Solution B until a maximum absorbance reading is obtained. Calculate the molar concentration of benzylpenicilloyl moiety in the Injection: Result = (10/3){[AM(3 + 0.02N)/3] AI}/MAB AM N = highest absorbance observed = number of 0.02-mL portions of Solution B added to the Sample solution to obtain the maximum absorbance = initial absorbance = molar absorptivity of the penamaldate formed by the reaction of benzylpenicilloyl with mercuric chloride at pH 7.6: 22,325
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USP 32
B = length of the cell (cm) Acceptance criteria: NLT 5.4 105 M and NMT 7.0 105 M SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 5833.0 USP Endotoxin Units/mL STERILITY TESTS 71: Meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration PH 791: 6.58.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, in a refrigerator. USP REFERENCE STANDARDS 11 USP Endotoxin RS
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% [NOTE2 g of specimen being used] HEAVY METALS, Method II 231: NMT 10 ppm SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 176182, with decomposition LOSS ON DRYING 731: Dry it in a vacuum over phosphorus pentoxide at 40 for 4 h: It loses NMT 0.2% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers
Beta Carotene
(Comment on this Monograph)id=m8730=Beta Carotene=BMonos.pdf)
Beta Carotene Capsules

(Comment on this Monograph)id=m8735=Beta Carotene Capsules=B-Monos.pdf) DEFINITION Beta Carotene Capsules contain NLT 90.0% and NMT 125.0% of the labeled amount of C40H56. IDENTIFICATION Grind a portion of the Capsule contents, equivalent to 10 mg of beta carotene, transfer to a centrifuge tube, add 5 mL of chloroform, shake for 1 min, and centrifuge for 3 min. Filter the supernatant layer, collecting 2 mL of the filtrate in a 25mL conical flask. Add 5 mL of antimony trichloride TS to the filtrate: A transient purple or blue color forms. ASSAY PROCEDURE [NOTEPerform this analysis in subdued light, using lowactinic glassware.] Cyclohexane: Spectrophotometric grade, or material that has been purified by being passed through a column of activated silica gel and distilled. Iodine solution: 0.01 mg/mL of iodine in cyclohexane. [NOTEPrepare this solution fresh daily.] Sample solution 1: Combine the contents of NLT 20 Capsules, and grind, using a freezer mill, to a fine powder of uniform color. Transfer a quantity of the finely ground Capsule contents, equivalent to 75 mg of beta carotene, to a 1000-mL volumetric flask, add 500 mL of water, and heat at 60 for 15 min. Cool to ambient temperature, and dilute with water to volume. Sample solution 2: Transfer 5.0 mL of Sample solution 1 to a glass-stoppered, 50-mL centrifuge tube. Add 3 g of sodium sulfate decahydrate, 2 mL of 1 N hydrochloric acid, and 20.0 mL of chloroform. Shake by mechanical means for 10 min, centrifuge for 5 min, and remove the aqueous layer without disturbing the chloroform layer. Add 2 g of anhydrous sodium sulfate to the chloroform layer, shake vigorously, and allow to settle. Sample solution 3: Transfer 5.0 mL of Sample solution 2 to a 50-mL volumetric flask, and add 30 mL of cyclohexane. Add 0.05 mL of Iodine solution, dilute with cyclohexane to volume, and allow to stand for 3 h. Transfer 20 mL of this solution to a centrifuge tube, and centrifuge for 2 min. Standard solution 1: 17 mg of beta carotene, previously subjected to the Assay and previously dried in vacuum over phosphorus pentoxide at 40 for 4 h, and transfer to a 1000-mL volumetric flask. Add 10 mL of water, heat at 60 for 15 min, and cool to room temperature. Add 3 g of sodium sulfate decahydrate and 2 mL of 1 N hydrochloric acid, and shake by mechanical means for 10 min. Add 200
536.87 C40H56 ,-Carotene; all-trans--Carotene; (all-E)-1,1-(3,7,12,16-Tetramethyl-1,3,5,7,9,11,13,15,17octadecanonaene-1,18-diyl)bis[2,6,6-trimethylcyclohexene] [7235-40-7]. DEFINITION Beta Carotene contains NLT 96.0% and NMT 101.0% of C40H56. IDENTIFICATION A. Determine the absorbances of the Assay Sample solution at 455 nm and at 483 nm: The ratio of the absorbance at 455 nm to that at 483 nm is 1.141.18. B. Determine the absorbance of the Assay Sample solution at 455 nm and that of the Assay Sample stock solution at 340 nm: The ratio of the absorbance at 455 nm to that at 340 nm is NLT 1.5. ASSAY PROCEDURE [NOTEPerform this procedure in subdued light, using lowactinic glassware.] Sample stock solution: Dissolve 50 mg of Beta Carotene in 10 mL of acid-free chloroform in a 100-mL volumetric flask. Dilute with cyclohexane to volume. Pipet 5 mL into a 100mL volumetric flask, and dilute with cyclohexane. Sample solution: Sample stock solution with cyclohexane (1 in 10) Spectrometric conditions Analytical wavelength: 455 nm Blank: Cyclohexane Sample: Sample solution Analysis: Determine the absorbance of Sample solution using the Blank. Calculate the percentage of C40H56 in the portion of Beta Carotene taken: Result = 400 (AU/250) 100 AU 250 = absorbance of the Sample solution = absorptivity of pure beta carotene
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mL of chloroform, and shake for 10 min. Add 750 mL of chloroform, shake, and dilute with chloroform to volume, disregarding the aqueous layer. Standard solution 2: Shake Standard solution 1 vigorously, and allow the layers to separate completely. Transfer 20 mL of the chloroform layer to a centrifuge tube, add 2 g of anhydrous sodium sulfate, shake vigorously, and allow to settle. Transfer 5.0 mL to a 50 mL volumetric flask, add 30 mL of cyclohexane. Add 0.05 mL of Iodine solution, dilute with cyclohexane to volume, and allow to stand for 3 h. Transfer 20 mL of Standard solution 2 to a centrifuge tube, and centrifuge for 2 min. Spectrometric conditions Analytical wavelength: 452 nm Blank: Cyclohexane Analysis Samples: Sample solution 3 and Standard solution 2 Measure the absorbances using the Blank. Calculate the percentage, of C40H56 in the portion of Capsule contents taken: Result = (AU/AS) (CS/CU) 100 = absorbance of Sample solution 3 = absorbance of Standard solution 2 = concentration of beta carotene in the Standard solution (g/mL) = nominal concentration of beta carotene in CU Sample solution 3 (g/mL) Acceptance criteria: 90.0%125.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements AU AS CS
Official Monographs / Betahistine 53

Standard solution: 0.38 mg/mL of USP Betahistine Hydrochloride RS in Mobile phase Sample solution: 0.38 mg/mL of Betahistine Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.0-mm 15-cm; packing L1 Temperature: 40 Flow rate: 0.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C8H12N2 2HCl in the portion of Betahistine Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Betahistine Hydrochloride RS in the Standard solution (mg/mL) = concentration of Betahistine Hydrochloride taken CU to prepare the Sample solution (mg/mL) Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Mobile phase and Chromatographic system: Proceed as directed in the Assay. Sample solution: 0.38 mg/mL of Betahistine Hydrochloride in Mobile phase Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Betahistine Hydrochloride taken: Result = (ru/rT) F 100 = peak response for each impurity = sum of the responses of all of the peaks, adjusted for the relative response factor F = response factor of the respective impurity (see Impurity Table) and 1.0 for all other peaks Acceptance criteria Individual impurities: See Impurity Table. Total impurities: NMT 0.5%
Impurity Table Relative Retention Time 0.3 0.4 Relative Response Factor 0.5 0.4 Acceptance Criteria, NMT (%) 0.2 0.2
rU rS CS
Betahistine Hydrochloride
(Comment on this Monograph)id=m8750=Betahistine Hydrochloride=B-Monos.pdf)
C8H12N2 2HCl 2-Pyridineethanamine, N-methyl-, dihydrochloride; 2-[2-(Methylamino)ethyl]pyridine; dihydrochloride [5579-84-0].
209.12
ru rT
DEFINITION Betahistine Hydrochloride contains NLT 99.0% and NMT 101.0% of C8H12N2 2HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.69 mg/mL of ammonium acetate in water, adjust with glacial acetic acid to a pH of 4.7 Mobile phase: Acetonitrile and Solution A (7:13), containing 2.88 mg/mL of sodium lauryl sulfate
Name 2-(2-Hydroxyethyl) pyridine 2-Vinylpyridine
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Betahistine / Official Monographs

Impurity Table (continued) Relative Retention Time 2.4 Relative Response Factor 1.4 Acceptance Criteria, NMT (%) 0.2
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Name N-Methyl-N,N-bis(2pyridin-2-yl-ethyl) amine Any other individual impurities
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS 231: NMT 10 ppm SPECIFIC TESTS PH 791: 0.81.2, in a solution (1 in 4) WATER DETERMINATION, Method I 921: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Betaine Hydrochloride RS
0.1
SPECIFIC TESTS PH 791: 2.03.0, in a solution (1 in 10) LOSS ON DRYING 731: Dry it between 100 and 105 to constant weight: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS USP REFERENCE STANDARDS 11 USP Betahistine Hydrochloride RS
Betamethasone
(Comment on this Monograph)id=m8760=Betamethasone=BMonos.pdf)
Betaine Hydrochloride
(Comment on this Monograph)id=m8755=Betaine Hydrochloride=B-Monos.pdf) 392.46 C22H29FO5 Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17,21-trihydroxy-16methyl-, (11,16)-; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione [378-44-9]. 153.61 C5H11NO2 HCl Methanaminium, 1-carboxy-N, N, N-trimethyl-, chloride; Betaine hydrochloride; (Carboxymethyl)trimethylammonium chloride [590-46-5]. DEFINITION Betaine Hydrochloride contains NLT 98.0% and NMT 100.5% of C5H11NO2 HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Chloride 191: mg/mL solution meets the requirements. DEFINITION Betamethasone contains NLT 97.0% and NMT 103.0% of C22H29FO5, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 0.5 mg/mL Betamethasone in dehydrated alcohol Developing solvent system: Chloroform and diethylamine (2:1) Analysis: Proceed as directed in the chapter, except to locate the spots by lightly spraying with dilute sulfuric acid (1 in 2) and heating on a hot plate or under a lamp until spots appear. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (37:63) Internal standard stock solution: 0.25 mg/mL propylparaben in alcohol Standard stock solution: 0.2 mg/mL USP Betamethasone RS in alcohol Standard solution: Internal standard stock solution and Standard stock solution (1:1)
A 50
ASSAY PROCEDURE Sample solution: Transfer 400 mg of Betaine Hydrochloride to a conical flask, add 50 mL of glacial acetic acid, and heat gently with swirling until solution is complete. Add 25 mL of mercuric acetate TS, cool, and add 2 drops of crystal violet TS. Analysis: Titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 15.36 mg of C5H11NO2 HCl.
USP 32
Sample stock solution: 0.2 mg/mL Betamethasone in alcohol Sample solution: Internal standard stock solution and Sample stock solution (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone and propylparaben are 1.0 and 1.4, respectively.] Suitability requirements Resolution: NLT 3.0 between betamethasone and propylparaben Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 taken: Result = (RU/RS) (CS/CU) 100 = peak height ratio of the betamethasone peak to the internal standard peak in the Sample solution = peak height ratio of the betamethasone peak to RS the internal standard peak in the Standard solution = concentration of USP Betamethasone RS in the CS Standard solution (mg/mL) = concentration of Betamethasone in the Sample CU solution (mg/mL) Acceptance criteria: 97.0%103.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2%, a platinum crucible being used Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Sample solution: Methanol Standard solution: Methanol Application volume: 10 L Eluant: Toluene, acetone, methyl ethyl ketone, and formic acid (55:20:20:5), in a nonequilibrated chamber Visualization: 5 SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +118 to +126, calculated on the dried basis Sample solution: 5 mg/mL, in methanol LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 2 and 30. USP REFERENCE STANDARDS 11 USP Betamethasone RS RU
Official Monographs / Betamethasone 55

IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 1 mg/mL, prepared by concentrating 10 mL of the Sample solution from the Assay on a steam bath to 1 mL Standard solution: 1 mg/mL of USP Betamethasone RS in dehydrated alcohol Developing solvent system: Chloroform and diethylamine (2:1) Spray reagent: Methanol, sulfuric acid, and nitric acid (10:10:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter, except to spray the plate with the Spray reagent, and heat at 105 for 10 min. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (37:63) Internal standard solution: 0.25 mg/mL of propylparaben in alcohol Standard stock solution: 0.2 mg/mL USP Betamethasone RS in alcohol Standard solution: Internal Standard solution and Standard stock solution (1:1) Sample solution: To a portion of Cream equivalent to 2 mg of betamethasone, add 10.0 mL of Internal standard solution and 10.0 mL of alcohol. Mix by rotation for 20 min. Centrifuge at 2500 rpm for 10 min. Transfer a portion of the supernatant to a suitable vial. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone and propylparaben are 1.0 and 1.4, respectively.] Suitability requirements Resolution: NLT 3.0 between betamethasone and propylparaben Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the label claim percentage of C22H29FO5 in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 = peak height ratio of the betamethasone peak to the internal standard peak from the Sample solution RS = peak height ratio of the betamethasone peak to the internal standard peak from the Standard solution = concentration of USP Betamethasone RS in the CS Standard solution (mg/mL) = nominal concentration of betamethasone in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for the absence of Staphylococcus aureus and Pseudomonas aeruginosa. RU
Betamethasone Cream
(Comment on this Monograph)id=m8770=Betamethasone Cream=B-Monos.pdf) DEFINITION Betamethasone Cream contains NLT 90.0% and NMT 115.0% of the labeled amount of C22H29FO5 in a suitable cream base.
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USP 32
rotate for 20 min. Centrifuge to clarify. Transfer 10-mL portions of the supernatant to separate, stoppered tubes. To each tube add 1 mL of blue tetrazolium TS, followed by 1 mL of Reagent, and mix. Heat in a 35 water bath for 1 h. Remove from the water bath, add 1 mL of glacial acetic acid to each tube, and mix. Cool to room temperature. Concomitantly determine the absorbances of the solutions in 1-cm cells at the wavelength of maximum absorbance at 525 nm, with a suitable spectrophotometer. Calculate the percentage of C22H29FO5 in each mL of the Oral Solution taken: Result = (AU AB)/(AS AB) (CS/CU) 100 absorbance of the Sample solution absorbance of the blank absorbance of the Standard solution concentration of USP Betamethasone RS in the Standard solution (mg/mL) = nominal concentration of betamethasone in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% AU AB AS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Store between 2 and 25, excursions permitted up to 30, protected from light. Preserve in a tight container. Protect from freezing. USP REFERENCE STANDARDS 11 USP Betamethasone RS = = = =
MINIMUM FILL 755:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight containers. USP REFERENCE STANDARDS 11 USP Betamethasone RS
Betamethasone Oral Solution

(Comment on this Monograph)id=m8780=Betamethasone Oral Solution=B-Monos.pdf) DEFINITION Betamethasone Oral Solution contains NLT 90.0 % and NMT 115.0 % of the labeled amount of betamethasone (C22H29FO5). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Evaporate 1 mL of the Sample solution, prepared as directed in the Assay, on a steam bath just to dryness, and dissolve the residue in 0.5 mL of alcohol. Developing solvent system: Chloroform and diethylamine (2:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter. Locate the spots by lightly spraying with dilute sulfuric acid (1 in 2) and heating on a hot plate or under a lamp until spots appear. ASSAY PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 0.6 mg/mL of USP Betamethasone RS in a mixture of chloroform and methanol (1:1) Sample solution: Equivalent to 1.2 mg of Betamethasone from Oral Solution in a 50-mL centrifuge tube. Rinse the pipet with 15 mL of 0.1 N hydrochloric acid, then with 20 mL of ethyl acetate, and add the rinsings to the centrifuge tube. Rotate for 10 min, or shake manually for 1 min. [NOTEDo not use a mechanical shaker.] Centrifuge to separate the phases. Transfer the upper phase (ethyl acetate) to a small, pear-shaped flask. Extract the aqueous phase twice more with 20-mL portions of ethyl acetate, and add the extracts to the pear-shaped flask. Evaporate the combined extracts on a steam bath under a gentle stream of nitrogen to dryness. Allow to cool to room temperature. Dissolve the residue in 0.5 mL of chloroform and methanol (1:1), using a vortex mixer. Transfer the solution to a 2-mL volumetric flask, with small portions of a mixture of chloroform and methanol (1:1), and dilute with the mixture of chloroform and methanol (1:1) to volume. Application volume: 200 L Developing solvent system: Chloroform, methanol, and ammonium hydroxide (175:20:1) Reagent: Tetramethylammonium hydroxide TS in alcohol (1 in 5) Analysis Samples: Sample solution and Standard solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow the spots to dry, and develop the chromatogram, using the Developing solvent system, until the solvent front has moved 15 cm. Remove the plates from the developing chamber, and allow them to dry for 15 min. Mark the betamethasone bands, using shortwavelength UV light, to include similar zones of silica gel for the Sample solution, the Standard solution, and a zone containing no betamethasone for the blank. Scrape off these zones, and transfer them to separate 50-mL centrifuge tubes. Add 15 mL of alcohol to each, and
Betamethasone Tablets
(Comment on this Monograph)id=m8790=Betamethasone Tablets=B-Monos.pdf) DEFINITION Betamethasone Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Evaporate 50 mL of the Sample solution, prepared as directed in the Assay, on a steam bath just to dryness, and dissolve the residue in 1 mL of chloroform. Developing solvent system: Chloroform and diethylamine (2:1) Application volume: 10 L Analysis Sample: Sample solution Proceed as directed in the chapter, except to locate the spots by lightly spraying with dilute sulfuric acid (1 in 2) and heating on a hot plate or under a lamp until spots appear. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (1:2) Internal standard solution: 0.125 mg/mL of beclomethasone in methanol Standard stock solution: 0.1 mg/mL of USP Betamethasone RS in methanol Standard solution: 0.05 mg/mL from a mixture of Standard stock solution and Internal standard solution (1:1) Sample solution: Equivalent to 0.5 mg of betamethasone from powdered Tablets (NLT 20 Tablets), into a 125-mL separator. Add 25 mL of water, and shake by mechanical means for 15 min. Add 5.0 mL of Internal standard solution. Extract with four 25-mL portions of chloroform. Filter the chloroform extracts through 4 g of chloroform-washed anhydrous sodium sulfate, collecting the extracts in a 150-
USP 32
mL beaker. Evaporate the extracts on a steam bath with the aid of a stream of nitrogen to dryness, taking care to avoid overheating. Dissolve the residue in 2 mL of methanol, and transfer to a 10-mL volumetric flask. Rinse the beaker with small portions of methanol, transferring the rinses to the same flask. Dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for beclomethasone and betamethasone are 1.4 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.7 between the analyte and internal standard peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak height ratio of the Sample solution = peak height ratio of the Standard solution = concentration of USP Betamethasone RS in the Standard solution (mg/mL) = nominal concentration of betamethasone in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU RS CS PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: 900 mL water Add 1.0 mL of Internal standard solution to each vessel. Apparatus 2: 50 rpm Time: 45 min Mobile phase: Methanol and water (3:2) Internal standard solution: 0.5 mg/mL of testosterone in methanol Standard stock solution: 0.5 mg/mL of USP Betamethasone RS in methanol Standard solution: 1 mL of Standard stock solution, and 1 mL of Internal standard solution. Dilute to 900 mL. Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 200 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone and testosterone are 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between betamethasone and testosterone Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity of C22H29FO5 dissolved in comparison with the Standard solution, similarly chromatographed.

Tolerances: NLT 75% (Q) of the labeled amount of C22H29FO5 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Standard solution: 12 g/mL instead of 10 g/mL of USP Betamethasone RS, as directed under Assay for Steroids 351 Sample solution: Weigh and finely powder 1 Tablet. Transfer to a 125-mL separator, add 20 mL of water, and shake. Extract the betamethasone completely, using three 15-mL portions of chloroform and filtering each extract through chloroform-washed cotton into a 50-mL volumetric flask. Dilute with chloroform to volume, and mix. Transfer 20.0 mL of this solution to a glass-stoppered, 50-mL conical flask, evaporate the chloroform on a steam bath just to dryness, cool, and dissolve the residue in 20.0 mL of alcohol. Analysis: Proceed as directed under Assay for Steroids 351, except to keep the flasks in a constant-temperature bath at 45 1 for 90 min, then add 1.0 mL of glacial acetic acid, and cool. Calculate the percentage of C22H29FO5 in the Tablet: (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Betamethasone RS in the Standard solution (g/mL) = concentration of the Sample solution, based the extent of dilution (g/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 2 and 25, excursions permitted between 15 and 30. [NOTEProtect the 21-tablet pack from excessive moisture.] USP REFERENCE STANDARDS 11 USP Betamethasone RS
Betamethasone Acetate
(Comment on this Monograph)id=m8820=Betamethasone Acetate=B-Monos.pdf) C24H31FO6 434.50 Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17-dihydroxy-16methyl-21-(acetyloxy)-, (11,16)-; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 21-acetate [987-24-6]. DEFINITION Betamethasone Acetate contains NLT 97.0% and NMT 103.0% of C24H31FO6, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 0.5 mg/mL in dehydrated alcohol Developing solvent system: Chloroform and diethylamine (2:1) Analysis: Proceed as directed in the chapter. Locate the spots on the plate by lightly spraying with 10% sulfuric acid in alcohol and heating on a hot plate or under a lamp until the spots appear. ASSAY PROCEDURE Mobile phase: (700:1.5:800)
Acetonitrile, glacial acetic acid, and water
58
USP 32
Internal standard solution: 0.7 mg/mL of progesterone in Mobile phase Standard stock solution: 0.5 mg/mL of USP Betamethasone Acetate RS in Mobile phase Standard solution: 10.0 mL Internal standard solution and 10.0 mL Standard stock solution. Dilute to 50 mL with Mobile phase. The concentration of USP Betamethasone Acetate RS is 0.1 mg/mL. Sample stock solution: 0.5 mg/mL of Betamethasone Acetate in Mobile phase Sample solution: 10.0 mL Internal standard solution and 10.0 mL Sample stock solution. Dilute to 50 mL with Mobile phase. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for progesterone and betamethasone acetate are 3 and 1.0, respectively.] Suitability requirements Resolution: NLT 2 between the analyte and internal standard peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C24H31FO6 in the portion taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the Sample solution = peak response ratio of the Standard solution = concentration of USP Betamethasone Acetate RS in the Standard solution (mg/mL) CU = concentration of Betamethasone in the Sample solution (mg/mL) Acceptance criteria: 97.0%103.0% RU RS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2%, a platinum crucible being used Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Sample solution: Methanol Standard solution: Methanol Application volume: 10 L Eluant: A mixture of toluene and isopropyl alcohol (90:10), in a nonequilibrated chamber Visualization: 5 SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +120 to +128 Sample solution: 10 mg/mL, in dioxane WATER DETERMINATION, Method I 921: NMT 4.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 2 and 30. USP REFERENCE STANDARDS 11 USP Betamethasone Acetate RS
Betamethasone Benzoate
(Comment on this Monograph)id=m8870=Betamethasone Benzoate=B-Monos.pdf) C29H33FO6 496.58 Pregna-1,4-diene-3,20-dione, 17-(benzoyloxy)-9-fluoro-11,21dihydroxy-16-methyl-, (11,16)-; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 17-benzoate [22298-29-9]. DEFINITION Betamethasone Benzoate contains NLT 98.0% and NMT 102.0% of C29H33FO6, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197M ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:2) Internal standard solution: 0.6 mg/mL of betamethasone dipropionate in methanol Standard stock solution: 0.6 mg/mL of USP Betamethasone Benzoate RS in methanol Standard solution: 0.2 mg/mL from Standard stock solution and Internal standard solution (1:2) Sample stock solution: 0.6 mg/mL of Betamethasone Benzoate in methanol Sample solution: 0.2 mg/mL from Sample stock solution and Internal standard solution (1:2) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 15 L System suitability Sample: Standard solution (three replicate injections) [NOTEThe retention times for betamethasone benzoate and betamethasone dipropionate are 5 min and 7 min, respectively.] Suitability requirements Resolution: NLT 3, between betamethasone benzoate and the internal standard Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C29H33FO6 in the portion of Betamethasone Benzoate taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = peak response ratios of the betamethasone benzoate peak to the internal standard peak from the Sample solution = peak response ratios of the betamethasone benzoate peak to the internal standard peak from the Standard solution = concentration of USP Betamethasone Benzoate RS in the Standard solution (mg/mL) = concentration of Betamethasone Benzoate in the Sample solution (mg/mL)
USP 32

Sample solution: Transfer the equivalent of 0.5 mg of betamethasone benzoate from the Gel to a 125-mL separatory funnel, add 20 mL of water and 2 mL of saturated sodium acetate solution, shake to disperse the Gel, and add 2 mL of Internal standard solution. Extract this solution with one 50-mL portion of chloroform followed by three 40-mL portions of chloroform. Discard the aqueous layer. Wash the chloroform extract with 10 mL of water, allow to stand for 10 min, then pass through chloroformwetted glass fiber filter paper and anhydrous sodium sulfate into a suitable container. Evaporate to dryness under a vacuum at 30. Dissolve the residue in 10 mL of methanol. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 236 nm Column: 3.9-mm 30-cm; packing L1 Temperature: 30 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for methyltestosterone and betamethasone benzoate are about 1.0 and 1.33, respectively.] Suitability requirements Resolution: NLT 3.0 between methyltestosterone and betamethasone benzoate Relative standard deviation: NMT 1.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage label claim of C29H33FO6 in the portion of Gel taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of betamethasone benzoate to methyltestosterone from the Sample solution = peak response ratio of betamethasone benzoate RS to methyltestosterone from the Standard solution CS = concentration of USP Betamethasone Benzoate RS in the Standard solution (g/mL) = nominal concentration of betamethasone CU benzoate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25, excursions permitted between 15 and 30. Protect from freezing. USP REFERENCE STANDARDS 11 USP Betamethasone Benzoate RS USP Methyltestosterone RS
IMPURITIES Organic Impurities PROCEDURE: RELATED STEROIDS Sample solution: 20.0 mg/mL in methanol Standard solution A: 5 mg/mL of USP Betamethasone Benzoate RS in methanol Standard solution B: 100 g/mL from Standard solution A in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Toluene, acetone, and methanol (75:25:4) Analysis Samples: Sample solution and Standard solutions Examine the plate under short-wavelength UV light. Acceptance criteria: The principal spot from the Sample solution corresponds in RF value to that of Standard solution A; and the Sample solution shows NMT 3 additional spots, the intensity and size of which do not exceed those of the spot from Standard solution B. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +60 to +66 Sample solution: 40 mg/mL, in dioxane LOSS ON DRYING 731: Dry 200 mg at 105 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store between 2 and 30. USP REFERENCE STANDARDS 11 USP Betamethasone Benzoate RS
Betamethasone Benzoate Gel

(Comment on this Monograph)id=m8890=Betamethasone Benzoate Gel=B-Monos.pdf) DEFINITION Betamethasone Benzoate Gel contains an amount of betamethasone benzoate (C29H33FO6) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone benzoate (C29H33FO6). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, acetonitrile, and water (23:9:18) Internal standard solution: 250 g/mL of USP Methyltestosterone RS in methanol Standard stock solution: 0.5 mg/mL of USP Betamethasone Benzoate RS in methanol Standard solution: 5.0 mL of Standard stock solution, 10 mL of Internal standard solution, diluted with methanol to 50 mL
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the peak obtained from the Internal standard in the Standard solution is 0.6 full-scale.] Detector: UV 254 nm or 240 nm Column: 4-mm 30-cm; packing L1 Temperature: Room temperature Column pressure: Capable up to 3500 psi Injection size: 5 L25 L System suitability Sample: Standard solution (three successive injections) Suitability requirements Relative standard deviation: NMT 2.0% between the lowest and highest peak area ratios Analysis Sample: Standard solution and Sample solution Determine the ratio of the peak heights, at equivalent retention times, obtained with the Sample solution and the Standard solution. Calculate the percentage of C28H37FO7 in the portion of Betamethasone Dipropionate taken: Result = (RU/RS) (CS/CU) 100 = peak height ratios of betamethasone dipropionate to the internal standard of the Sample solution = peak height ratios of betamethasone RS dipropionate to the internal standard of the Standard solution CS = concentration of USP Betamethasone Dipropionate RS in the Standard solution (mg/mL) CU = nominal concentration of Betamethasone Dipropionate in the Sample solution (mg/mL) Acceptance criteria: 97.0%103.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2%, a platinum crucible being used Organic Impurities PROCEDURE Mobile phase: Acetonitrile and water (13:7) System suitability solution: 0.05 mg/mL of each USP Betamethasone Dipropionate RS and USP Betamethasone Valerate RS in Mobile phase Sample solution: 0.3 mg/mL of Betamethasone Dipropionate in Mobile phase, and shake until dissolved Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 4.0 between betamethasone valerate and betamethasone dipropionate RU
Betamethasone Dipropionate
(Comment on this Monograph)id=m8960=Betamethasone Dipropionate=B-Monos.pdf)
504.60 C28H37FO7 Pregna-1,4-diene-3,20-dione, 9-fluoro-11-hydroxy-16methyl-17,21-bis(1-oxopropoxy)-, (11,16); 9-Fluoro-11,17,21-trihydroxy-16methylpregna-1,4-diene-3,20-dione 17,21-dipropionate [5593-20-4]. DEFINITION Betamethasone Dipropionate contains NLT 97.0% and NMT 103.0% of C28H37FO7, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 1 mg/mL, in chloroform Developing solvent system: Chloroform and acetone (7:1) ASSAY PROCEDURE Mobile phase: Acetonitrile solution (1 in 2), degassed by ultrasonic vibration for 5 to 10 min, such that the retention time of betamethasone dipropionate is approximately 14 min and that of beclomethasone dipropionate is approximately 18 min. [NOTEDo not leave the Mobile phase in the column overnight, but flush the system after use with water for 15 min, followed by methanol for 15 min.] Internal standard solution: 0.9 mg/mL of USP Beclomethasone Dipropionate RS in a solution of acetic acid in methanol (1 in 1000) Standard stock solution: 0.6 mg/mL of USP Betamethasone Dipropionate RS in a solution of acetic acid in methanol (1 in 1000) Standard solution: Standard stock solution and Internal standard solution (1:1) [NOTEThe concentrations for the Standard solution are 0.3 mg/mL of betamethasone dipropionate and 0.45 mg/mL of beclomethasone dipropionate.] Sample stock solution: 0.6 mg/mL of Betamethasone Dipropionate in a solution of acetic acid in methanol (1 in 1000) Sample solution: Sample stock solution and Internal standard solution (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC [NOTESeparately inject equal volumes (5 L25 L) of the Sample solution and Standard solution by means of a suitable microsyringe or sampling valve, adjusting the specimen size and other operating parameters such that
USP 32
Column efficiency: NLT 8000 theoretical plates Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Betamethasone Dipropionate taken: Result = (ru/rT) 100 = peak response for each impurity ru = sum of the responses for all the peaks rT Acceptance criteria Individual impurities: NMT 1.0% of any individual impurity is found Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +63 to +70 Sample solution: 10 mg/mL, in dioxane LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS USP Betamethasone Dipropionate RS USP Betamethasone Valerate RS

Diluent: Isopropyl alcohol containing acetic acid (1 in 1000) Internal standard solution: 0.90 mg/mL of USP Beclomethasone Dipropionate RS in Diluent Standard stock solution: 0.642 mg/mL of USP Betamethasone Dipropionate RS in Diluent Standard solution: 10.0 mL Standard stock solution and 10.0 mL Internal standard solution, dilute with Diluent to 100 mL [NOTEThe concentrations for the Standard solution are 0.09 mg/mL of beclomethasone dipropionate and 0.0642 mg/mL of betamethasone dipropionate.] Sample solution: Discharge the entire contents of the container of Topical Aerosol into a 100-mL volumetric flask. Allow the solution to warm to room temperature slowly to prevent it from boiling out of the flask, then evaporate the propellant by swirling the flask in a water bath at 25 until the solution stops bubbling. Add 10.0 mL of Internal standard solution, and dilute with Diluent to volume. Pass the solution through a 0.45-m filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC [NOTESeparately inject equal volumes (525 L) of the Standard solution and Sample solution by means of a suitable microsyringe or sampling valve, adjusting the specimen size and other operating parameters such that the peak obtained from the internal standard in the Standard solution is 0.6 full-scale.] Detector: UV 254 nm or 240 nm Column: 4-mm 30-cm; packing L1 Temperature: Room temperature Column pressure: Capable up to 3500 psi Injection size: 525 L System suitability Sample: Standard solution [NOTEthree successive injections] Suitability requirements Relative standard deviation: NMT 2.0% between the lowest and highest peak area ratios Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 equivalent to the quantity of C28H37FO7 in the container of the Topical Aerosol: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak height ratios of the betamethasone dipropionate to beclomethasone dipropionate peaks from the Sample solution RS = peak height ratios of the betamethasone dipropionate to beclomethasone dipropionate peaks from the Standard solution CS = concentration of USP Betamethasone Dipropionate RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Mr1 = molecular weight of betamethasone, 392.46 Mr2 = molecular weight of betamethasone dipropionate, 504.60 Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS OTHER REQUIREMENTS: It meets the requirements for Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers 601, Pressure Test, Minimum Fill, and Leakage Test.
Betamethasone Dipropionate Topical Aerosol

(Comment on this Monograph)id=m8970=Betamethasone Dipropionate Topical Aerosol=B-Monos.pdf) DEFINITION Betamethasone Dipropionate Topical Aerosol is a solution, in suitable propellants in a pressurized container, of betamethasone dipropionate (C28H37FO7) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 3.2 mg/mL of USP Betamethasone Dipropionate RS in methanol Sample solution: Place the container in a dry icemethanol bath for 5 min. Open the can by means of a tube-cutter, and allow the propellant to evaporate under a gentle stream of nitrogen for 1 h. Transfer 3 mL of the residue to a 50-mL centrifuge tube. Add 10 mL of a mixture of methanol and water (4:1), and shake vigorously. Centrifuge to clarify. Application volume: 25 L Developing solvent system: Toluene and ethyl acetate (1:1) Analysis Samples: Standard solution and Sample solution Proceed as directed in the chapter. Spray the plate with a mixture of sulfuric acid, methanol, and nitric acid (10:10:1), and heat at 105 for 15 min. ASSAY PROCEDURE Mobile phase: Acetonitrile solution (1 in 2), degassed by ultrasonic vibration for 510 min, such that the retention time of betamethasone dipropionate is approximately 14 min and that of beclomethasone dipropionate is approximately 18 min. [NOTEDo not leave the Mobile phase in the column overnight, but flush the system after use with water for 15 min, followed by methanol for 15 min.]
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for 5 min. Transfer a portion of the supernatant to a suitable vial. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC [NOTESeparately inject equal volumes (5 L25 L) of the Sample solution and Standard solution by means of a suitable microsyringe or sampling valve, adjusting the specimen size and other operating parameters such that the peak obtained from the Internal standard solution in the Standard solution is 0.6 full-scale.] Detector: UV 254 nm or 240 nm Column: 4-mm 30-cm; packing L1 Temperature: Room temperature Column pressure: Capable up to 3500 psi Injection size: 5 L25 L System suitability Sample: Standard solution (three successive injections) Suitability requirements Relative standard deviation: NMT 2.0% between the lowest and highest peak area ratios Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H29FO5 in the portion of Cream taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak height ratio of the betamethasone dipropionate peak and the internal standard peak obtained from the Sample solution = peak height ratio of the betamethasone RS dipropionate peak and the internal standard peak obtained from the Standard solution = concentration of USP Betamethasone CS Dipropionate RS in the Standard solution (mg/mL) = nominal concentration of betamethasone CU dipropionate in the Sample solution (mg/mL) = molecular weight of betamethasone, 392.46 Mr1 = molecular weight of betamethasone Mr2 dipropionate, 504.60 Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers. Store at 25; excursions permitted between 15 and 30. Protect from freezing. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS USP Betamethasone Dipropionate RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, pressurized containers, and avoid exposure to excessive heat. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS USP Betamethasone Dipropionate RS
Betamethasone Dipropionate Cream

(Comment on this Monograph)id=m8980=Betamethasone Dipropionate Cream=B-Monos.pdf) DEFINITION Betamethasone Dipropionate Cream contains an amount of betamethasone dipropionate (C28H37FO7) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5), in a suitable cream base. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 150 g/mL of USP Betamethasone Dipropionate RS in chloroform Sample solution: 1.5 g of Cream to a glass-stoppered, 50mL centrifuge tube. Add 15 mL of a methanolhydrochloric acid solution prepared by mixing 1 volume of dilute hydrochloric acid (1 in 120) with 4 volumes of methanol. Shake to obtain a homogeneous mixture. Add 30 mL of solvent hexane, mix for 10 min, and centrifuge. Using a suitable syringe, transfer the lower aqueous phase to a second centrifuge tube, add 20 mL of water. Extract this aqueous mixture with chloroform by shaking, centrifuging, and removing the lower, chloroform phase with a syringe. Evaporate the chloroform on a steam bath with the aid of a stream of nitrogen to dryness, cool, and dissolve the residue in chloroform to obtain a solution containing 150 g/mL of betamethasone dipropionate. Application volume: 40 L Developing solvent system: Chloroform and acetone (7:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter. ASSAY PROCEDURE Mobile phase: Acetonitrile solution (1 in 2), degassed by ultrasonic vibration for 5 to 10 min, such that the retention time of betamethasone dipropionate is approximately 14 min and that of beclomethasone dipropionate is approximately 18 min. [NOTEDo not leave the Mobile phase in the column overnight, but flush the system after use with water for 15 min, followed by methanol for 15 min.] Diluent: Acetic acid in methanol (1 in 1000) Internal standard solution: 0.45 mg/mL of USP Beclomethasone Dipropionate RS in Diluent Standard stock solution: 0.2 mg/mL of USP Betamethasone Dipropionate RS in Diluent Standard solution: Standard stock solution and Internal standard solution (2:1) [NOTEThe concentrations for the Standard solution are 0.133 mg/mL of betamethasone dipropionate and 0.15 mg/mL of beclomethasone dipropionate.] Sample solution: Equivalent to 2 mg of betamethasone dipropionate from Cream, into a capped 50-mL centrifuge tube. Add 5.0 mL of Internal standard solution and 10.0 mL of Diluent. Heat in a water bath at 60, shaking intermittently, until the specimen melts. Remove from the bath, and shake vigorously until the specimen has resolidified. Repeat the heating and shaking. Freeze in an icemethanol bath for 15 min, and centrifuge at 2500 rpm
Betamethasone Dipropionate Lotion

(Comment on this Monograph)id=m8990=Betamethasone Dipropionate Lotion=B-Monos.pdf) DEFINITION Betamethasone Dipropionate Lotion contains an amount of betamethasone dipropionate (C28H37FO7) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5), in a suitable lotion base. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 150 g/mL of USP Betamethasone Dipropionate RS in chloroform
USP 32
Sample solution: Equivalent to 0.6 mg of betamethasone dipropionate from Lotion. In a 50-mL vial, add 10 mL of 0.1 N hydrochloric acid, then add 4 mL of chloroform. Disperse on a vortex mixer for about 1 min, then shake vigorously for 10 min, and centrifuge at 2000 rpm for about 5 min. Transfer the chloroform layer to a suitable vial. Application volume: 40 L Developing solvent system: Chloroform and acetone (7:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter. ASSAY PROCEDURE Mobile phase: Acetonitrile solution (about 1 in 2), degassed by ultrasonic vibration for 5 to 10 min, such that the retention time of betamethasone dipropionate is approximately 14 min and that of beclomethasone dipropionate is approximately 18 min [NOTEDo not leave the Mobile phase in the column overnight, but flush the system after use with water for 15 min, followed by methanol for 15 min.] Internal standard solution: 0.9 mg/mL of USP Beclomethasone Dipropionate RS in chloroform Standard stock solution: 0.6 mg/mL of USP Betamethasone Dipropionate RS in chloroform Standard solution: Standard stock solution and Internal standard solution (1:1) [NOTEThe concentrations for the Standard solution are 0.3 mg/mL of betamethasone dipropionate and 0.45 mg/mL of beclomethasone dipropionate.] To 10.0 mL of 0.1 N hydrochloric acid in a capped 5-mL centrifuge tube add 4.0 mL of the solution, and treat this solution as follows. Cap, and shake vigorously for about 2 min, or disperse on a vortex mixer for about 1 min. Centrifuge at 2500 rpm for about 3 min. Transfer the chloroform phase to a suitable vial. Evaporate the chloroform under a stream of nitrogen at a slightly elevated temperature to dryness. Cool the vial to room temperature, add 4.0 mL of methanol, and swirl to dissolve the residue. Sample solution: Equivalent to 1.2 mg of betamethasone dipropionate from Lotion. In a capped 50-mL centrifuge tube, add 10.0 mL of 0.1 N hydrochloric acid, shake to disperse, then add 2.0 mL of Internal standard solution and 2.0 mL of chloroform. Cap, and shake vigorously for about 2 min, or disperse on a vortex mixer for about 1 min. Centrifuge at 2500 rpm for about 3 min. Transfer the chloroform phase to a suitable vial. Evaporate the chloroform under a stream of nitrogen at a slightly elevated temperature to dryness. Cool the vial to room temperature, add 4.0 mL of methanol, and swirl to dissolve the residue. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 or 240 nm Column: 4-mm 30-cm; packing L1 Temperature: Room temperature Column pressure: Capable up to 3500 psi Injection size: 525 L System suitability Samples: Standard solution [NOTEthree successive injections] Suitability requirements Relative standard deviation: NMT 2.0% between the lowest and highest peak area ratios Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H29FO5 in the portion of Lotion taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 RU

= peak height ratios of the betamethasone dipropionate peak to the internal standard peak from the Sample solution = peak height ratios of the betamethasone RS dipropionate peak to the internal standard peak from the Standard solution = concentration of USP Betamethasone CS Dipropionate RS in the Standard solution (mg/mL) = nominal concentration of betamethasone CU dipropionate in the Sample solution (mg/mL) = molecular weight of betamethasone, 392.46 Mr1 = molecular weight of betamethasone Mr2 dipropionate, 504.60 Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MINIMUM FILL 755: Meets the requirements
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25; excursions permitted between 15 and 30. Protect from light and freezing. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS USP Betamethasone Dipropionate RS
Betamethasone Dipropionate Ointment

(Comment on this Monograph)id=m9020=Betamethasone Dipropionate Ointment=B-Monos.pdf) DEFINITION Betamethasone Dipropionate Ointment contains an amount of betamethasone dipropionate (C28H37FO7) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5), in a suitable ointment base. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 150 g/mL of USP Betamethasone Dipropionate RS in chloroform Sample solution: 1.5 g of Ointment to a glass-stoppered, 50-mL centrifuge tube. Add 15 mL of methanolhydrochloric acid solution prepared by mixing 1 volume of dilute hydrochloric acid (1 in 120) with 4 volumes of methanol. Shake to obtain a homogeneous mixture. Add 30 mL of solvent hexane, mix for 10 min, and centrifuge. Using a suitable syringe, transfer the lower aqueous phase to a second centrifuge tube, add 20 mL of water, and mix. Extract this aqueous mixture with chloroform by shaking, centrifuging, and removing the lower, chloroform phase with a syringe. Evaporate the chloroform on a steam bath with the aid of a stream of nitrogen to dryness, cool, and dissolve the residue in chloroform to obtain a solution containing 150 g/mL of betamethasone dipropionate. Application volume: 40 L Developing solvent system: Chloroform and acetone (7:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter. ASSAY PROCEDURE Mobile phase: Acetonitrile solution (1 in 2), degassed by ultrasonic vibration for 510 min, such that the retention time of betamethasone dipropionate is approximately 14 min and that of beclomethasone dipropionate is approximately 18 min. [NOTEDo not leave the Mobile phase in the column overnight, but flush the system after
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USP Betamethasone Dipropionate RS
USP 32
use with water for 15 min, followed by methanol for 15 min.] Diluent: Acetic acid in alcohol (1 in 1000) Internal standard solution: 0.45 mg/mL of USP Beclomethasone Dipropionate RS in Diluent Standard stock solution: 0.2 mg/mL of USP Betamethasone Dipropionate RS in Diluent Standard solution: Standard stock solution and Internal standard solution (2:1) [NOTEThe concentrations for the Standard solution are 0.133 mg/mL of betamethasone dipropionate and 0.15 mg/mL of beclomethasone dipropionate.] Sample solution: Equivalent to 2 mg of betamethasone dipropionate from Ointment, in a capped 50-mL centrifuge tube. Add 5.0 mL of Internal standard solution and 10.0 mL of Diluent. Heat in a water bath at 70, shaking intermittently until the sample melts. Remove from the bath, and shake vigorously until the ointment has solidified. Repeat the heating and shaking operation. Freeze in an icemethanol bath for 15 min, and centrifuge at 2500 rpm for 5 min. Transfer a portion of the supernatant to a suitable vial. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC [NOTEUse a suitable microsyringe or sampling valve, and adjust the specimen size and other operating parameters such that the peak obtained from the internal standard in the Standard solution is 0.6 full-scale.] Detector: UV 254 nm or 240 nm Column: 4-mm 30-cm; packing L1 Temperature: Room temperature Column pressure: Capable up to 3500 psi Injection size: 525 L System suitability Sample: Standard solution [NOTEthree successive injections] Suitability requirements Relative standard deviation: NMT 2.0% between the lowest and highest peak area ratios Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H29FO5 in the portion of Ointment: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak height ratio of the betamethasone dipropionate peak and the internal standard peak from the Sample solution RS = peak height ratio of the betamethasone dipropionate peak and the internal standard peak from the Standard solution CS = concentration of USP Betamethasone Dipropionate RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Mr1 = molecular weight of betamethasone, 392.46 Mr2 = molecular weight of betamethasone dipropionate, 504.60 Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers. Store at 25, excursions permitted between 15 and 30. Protect from freezing. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS
Betamethasone Sodium Phosphate

(Comment on this Monograph)id=m9070=Betamethasone Sodium Phosphate=B-Monos.pdf) C22H28FNa2O8P 516.40 Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17-dihydroxy-16methyl-21-(phosphonooxy)-, disodium salt, (11,16)-; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 21-(disodium phosphate) [151-73-5]. DEFINITION Betamethasone Sodium Phosphate contains NLT 97.0% and NMT 103.0% of C22H28FNa2O8P, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 1 mg/mL of USP Betamethasone Sodium Phosphate RS in methanol Sample solution: 1 mg/mL Developing solvent system: 500 mL of butyl alcohol and 200 mL of dilute hydrochloric acid (1 in 12) Place in a separatory funnel, and mix. Use the organic layer as the developing solvent. Spray reagent: A mixture of sulfuric acid, methanol, and nitric acid (10:10:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in Thin-Layer Chromatographic Identification Test 201, except to spray the plate with Spray reagent, and heat at 105 for 10 min. C. IDENTIFICATION TESTSGENERAL, Sodium 191 AND Phosphate 191: Ignite it at 800 (see Residue on Ignition 281): the residue meets the requirements. ASSAY PROCEDURE Mobile phase: Methanol and 0.07 M anhydrous monobasic potassium phosphate (3:2) Standard solution: 0.17 mg/mL of USP Betamethasone Sodium Phosphate RS in a mixture of methanol and water (3:2) Sample solution: 0.17 mg/mL of Betamethasone Sodium Phosphate in a mixture of methanol and water (3:2) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 3.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H28FNa2O8P in the portion of Betamethasone Sodium Phosphate taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
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= concentration of USP Betamethasone Sodium Phosphate RS in the Standard solution (mg/mL) = concentration of Betamethasone Sodium CU Phosphate in the Sample solution (mg/mL) Acceptance criteria: 97.0%103.0% IMPURITIES Inorganic Impurities LIMIT OF PHOSPHATE IONS Standard phosphate solution and Phosphate reagent A: Prepare as directed under Phosphate in Reagents (see Reagents, Indicators, and SolutionsGeneral Tests for Reagents). Phosphate reagent B: 350 mg of p-methylaminophenol sulfate in 50 mL of water. Add 20 g of sodium metabisulfite, mix to dissolve, and dilute with water to 100 mL. Sample solution: 50 mg of Betamethasone Sodium Phosphate in a mixture of 10 mL of water and 5 mL of 2 N sulfuric acid contained in a 25-mL volumetric flask, by warming if necessary. Add 1 mL each of Phosphate reagent A and Phosphate reagent B, dilute with water to 25.0 mL, mix, and allow to stand at room temperature for 30 min. Standard solution: 5.0 mL of Standard phosphate solution in a mixture of 10 mL of water and 5 mL of 2 N sulfuric acid contained in a 25-mL volumetric flask, by warming if necessary. Add 1 mL each of Phosphate reagent A and Phosphate reagent B, dilute with water to 25.0 mL, mix, and allow to stand at room temperature for 30 min. Blank: Water Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 730 nm Cell: 1 cm Analysis Samples: Sample solution and Standard solution Acceptance criteria: The absorbance of the Sample solution is NMT that of the Standard solution. The limit is 1.0% of phosphate (PO4). Organic Impurities PROCEDURE: LIMIT OF FREE BETAMETHASONE Sample solution: 25.0 mg of Betamethasone Sodium Phosphate in water to make 25.0 mL. Transfer 5.0 mL of the solution to a separator, and extract with three 25-mL portions of chloroform. Filter each extract through a chloroform-saturated cotton pledget, combining the filtrates in a conical flask. Evaporate the chloroform on a steam bath with the aid of a current of air to dryness, and dissolve the residue in methanol to make 25.0 mL. Blank solution: 5.0 mL of water transferred to a separator Proceed as directed under Sample solution. The methanolic solution so obtained is the Blank solution. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 239 nm Blank: Blank solution Cell: 1 cm Analysis Sample: Sample solution Calculate the quantity, in mg, of free betamethasone in the portion of Betamethasone Sodium Phosphate taken: Result = A 3.125 CS

A = absorbance of the Sample solution Acceptance criteria: 250 g (1.0%) SPECIFIC TESTS SPECIFIC ROTATION 781S: +99 to +105 Sample solution: 10 mg/mL WATER DETERMINATION, Method I 921: NMT 10.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Betamethasone Sodium Phosphate RS
Betamethasone Sodium Phosphate Injection

(Comment on this Monograph)id=m9080=Betamethasone Sodium Phosphate Injection=B-Monos.pdf) DEFINITION Betamethasone Sodium Phosphate Injection is a sterile solution of Betamethasone Sodium Phosphate in Water for Injection. It contains an amount of betamethasone sodium phosphate (C22H28FNa2O8P) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: Chromatographic silica gel mixture Standard solution: 2 mg/mL of USP Betamethasone Sodium Phosphate RS in methanol Sample solution: 2 mg/mL of betamethasone sodium phosphate from Injection in methanol Application volume: 10 L Analysis Samples: Sample solution and Standard solution Develop the chromatogram in an equilibrated chamber containing n-butyl alcohol previously shaken with 1 N hydrochloric acid, until the solvent front has moved threefourths of the length of the plate. Remove the plate from the developing chamber, air-dry, then spray with a mixture of sulfuric acid, methanol, and nitric acid (10:10:1). Heat the plate at 105 for 10 min. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol and 0.05 M monobasic potassium phosphate (1:1) Internal standard solution: 1 mg/mL of butylparaben in methanol Standard stock solution: 4 mg/mL of USP Betamethasone Sodium Phosphate RS Standard solution: 3.0 mL of Standard stock solution and 5.0 mL of Internal standard solution. Dilute to 25 mL. Sample solution: Equivalent to 9 mg of betamethasone from a volume of Injection, in a 25-mL volumetric flask. Add 5.0 mL of the Internal standard solution, and dilute to volume. Chromatographic system (See Chromatography 621, System Suitability.)
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IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Absorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 2 mg/mL of USP Betamethasone Sodium Phosphate RS in methanol and water solution (1:1) Sample solution: Dilute 2 mL with 2 mL of methanol. Application volume: 10 L Developing solvent system: Place 500 mL of butyl alcohol and 200 mL of dilute hydrochloric acid (1 in 12) in a separatory funnel, and mix. Use the organic layer as the developing solvent. Spray reagent: A mixture of sulfuric acid, methanol, and nitric acid (10:10:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter except to spray the plate with Spray reagent, and heat at 105 for 10 min. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Absorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 1.5 mg/mL of USP Betamethasone Acetate RS in methanol and water solution (1:1) Sample solution: Dilute 2 mL with 2 mL of methanol. Application volume: 10 L Developing solvent system: Chloroform and diethylamine (2:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter, except to locate the spots by lightly spraying with dilute sulfuric acid (1 in 2), and heating on a hot plate or under a lamp until spots appear. ASSAY PROCEDURE Mobile phase: Methanol and 0.075 M monobasic potassium phosphate (7:5) Internal standard solution: 1 mg/mL of methyltestosterone in methanol Standard stock solution A: 2.52 mg/mL of USP Betamethasone Sodium Phosphate RS in Mobile phase Standard stock solution B: 1.8 mg/mL of USP Betamethasone Acetate RS in methanol Standard solution: 5 mL Standard stock solution A, 5 mL Standard stock solution B, 10 mL Internal Standard solution. Dilute to 100 mL with Mobile phase. [NOTEThe Standard solution has a known concentration of 126 g/mL USP Betamethasone Sodium Phosphate RS and 90 g/mL USP Betamethasone Acetate RS.] Sample solution: Equivalent to 9 mg of betamethasone acetate. Using a To contain pipet, transfer a volume of the well-mixed Injectable Suspension to a 100-mL volumetric flask. Rinse the pipet with 10 mL of Mobile phase, collecting the rinse in the volumetric flask. Add 10 mL of Internal standard solution, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone phosphate, methyltestosterone, and betamethasone acetate are 0.5, 1.7, and 1.0, respectively.]
Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone sodium phosphate and butylparaben are 1.0 and 2.4, respectively.] Suitability requirements Resolution: NLT 5 between the analyte and internal standard peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in each mL of Injection taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratio of the Sample solution = peak response ratio of the Standard solution = concentration of USP Betamethasone Sodium Phosphate RS in the Standard solution (mg/mL) = nominal concentration of betamethasone in the CU Sample solution (mg/mL) = molecular weight of betamethasone, 392.47 Mr1 = molecular weight of betamethasone sodium Mr2 phosphate, 516.41 Acceptance criteria: 90.0%110.0% RU RS CS SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 29.2 USP Endotoxin Units/mg of betamethasone PH 791: 8.09.0 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Betamethasone Sodium Phosphate RS USP Endotoxin RS
Betamethasone Sodium Phosphate and Betamethasone Acetate Injectable Suspension

(Comment on this Monograph)id=m9125=Betamethasone Sodium Phosphate and Betamethasone Acetate Injectable Suspension=B-Monos.pdf) DEFINITION Betamethasone Sodium Phosphate and Betamethasone Acetate Injectable Suspension is a sterile solution of Betamethasone Sodium Phosphate in solution and Betamethasone Acetate in suspension in Water for Injection. It contains an amount of betamethasone sodium phosphate (C22H28FNa2O8P) equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of betamethasone (C22H29FO5), and NLT 90.0% and NMT 115.0% of the labeled amount of betamethasone acetate (C24H31FO6).
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Suitability requirements Resolution: NLT 5.0 between the betamethasone phosphate and betamethasone acetate peaks; NLT 3.0 between the betamethasone acetate and internal standard peaks Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C24H31FO6 in each mL of Injectable Suspension taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio for betamethasone acetate and methyltestosterone from the Sample solution RS = peak response ratio for betamethasone acetate and methyltestosterone from the Standard solution CS = concentration of USP Betamethasone Acetate RS in the Standard solution (g/mL) CU = nominal concentration of the Sample solution (mg/mL) Calculate the percentage of C22H29FO5 equivalent to the quantity of C22H28FNa2O8P in each mL of Injectable Suspension taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratio for betamethasone phosphate and methyltestosterone from the Sample solution = peak response ratio for betamethasone RS phosphate and methyltestosterone from the Standard solution = concentration of USP Betamethasone Sodium CS Phosphate RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of betamethasone, 392.46 Mr1 = molecular weight of betamethasone sodium Mr2 phosphate, 516.41 Acceptance criteria: 90.0%115.0% RU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 29.2 USP Endotoxin Units/mg of betamethasone PH 791: 6.87.2 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Betamethasone Acetate RS RU

USP Betamethasone Sodium Phosphate RS USP Endotoxin RS
Betamethasone Valerate
(Comment on this Monograph)id=m9180=Betamethasone Valerate=B-Monos.pdf)
C27H37FO6 476.59 Pregna-1,4-diene-3,20-dione, 9-fluoro-11,21-dihydroxy-16methyl-17-[(1-oxopentyl)oxy]-, (11,16)-; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 17-valerate [2152-44-5]. DEFINITION Betamethasone Valerate contains NLT 97.0% and NMT 103.0% of C27H37FO6, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 1 mg/mL, in alcohol Developing solvent system: Toluene and ethyl acetate (1:1) Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter. Spray the plate with a mixture of sulfuric acid, methanol, and nitric acid (10:10:1), and heat at 105 for 15 min. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:2) Diluent: Glacial acetic acid in methanol (1 in 1000) Internal standard solution: 0.4 mg/mL of USP Beclomethasone Dipropionate RS in Diluent Standard stock solution: 0.6 mg/mL of USP Betamethasone Valerate RS in Diluent Standard solution: Standard stock solution and Internal standard solution (1:2) [NOTEThis solution has a concentration of 0.2 mg/mL of USP Betamethasone Valerate RS.] Sample stock solution: 0.6 mg/mL of Betamethasone Valerate in Diluent Sample solution: Sample stock solution and Internal standard solution (1:2) Chromatographic system (See Chromatography 621, System Suitability.)
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS USP Betamethasone Valerate RS
Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone valerate and beclomethasone dipropionate are 1.0 and 1.7, respectively.] Suitability requirements Resolution: NLT 4.5 between betamethasone valerate and beclomethasone dipropionate Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C27H37FO6 in the portion of Betamethasone Valerate taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the Sample solution = peak response ratio of the Standard solution = concentration of USP Betamethasone Valerate RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 97.0%103.0% RU RS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2%, using a platinum crucible Organic Impurities PROCEDURE Mobile phase: Acetonitrile, glacial acetic acid, and water (550:1:450) Sample solution: 0.4 mg/mL of Betamethasone Valerate in Mobile phase. Shake until dissolved. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Sample solution Suitability requirements Resolution: NLT 1.5 between betamethasone valerate and any impurity Column efficiency: NLT 9000 theoretical plates Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Betamethasone Valerate taken: Result = (rU/rT) 100 = peak response for each impurity rU rT = sum of all the peak responses Acceptance criteria Individual impurities: NMT 1.0% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +75 to +82 Sample solution: 10 mg/mL, in dioxane LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 0.5% of its weight.
Betamethasone Valerate Cream

(Comment on this Monograph)id=m9200=Betamethasone Valerate Cream=B-Monos.pdf) DEFINITION Betamethasone Valerate Cream contains an amount of betamethasone valerate (C27H37FO6) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5), in a suitable cream base. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Transfer the equivalent to 2 mg of betamethasone from Cream to a separator, add 20 mL of water and 2 mL of dilute hydrochloric acid (1 in 120), and mix. Extract with four 50-mL portions of chloroform, and combine the extracts. Filter through a cotton pledget, previously layered over with anhydrous sodium sulfate. Evaporate the filtrates on a steam bath under a stream of dry nitrogen to dryness. Dissolve the residue in alcohol to obtain a solution containing about 1 mg/mL. Developing solvent system: Toluene and ethyl acetate (1:1) Application volume: 10 L Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter. Spray the plate with a mixture of sulfuric acid, methanol, and nitric acid (10:10:1), and heat at 105 for 15 min. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:2) Diluent: Glacial acetic acid in methanol (1 in 1000) Internal standard solution: 0.4 mg/mL of USP Beclomethasone Dipropionate RS in Diluent Standard stock solution: 0.6 mg/mL of USP Betamethasone Valerate RS in Diluent Standard solution: Standard stock solution and Internal standard solution (1:2) [NOTEThis solution has a concentration of 0.2 mg/mL of USP Betamethasone Valerate RS.] Sample solution: Equivalent to 2.5 mg of betamethasone from Cream, in a 50-mL centrifuge tube. Add 10.0 mL of the Internal standard solution and 5.0 mL of Diluent. Insert the stopper into the tube, and place in a water bath held at 60 until the specimen melts. Remove from the bath, and shake vigorously until the specimen resolidifies. Repeat the heating and shaking two more times. Place the tube in an icemethanol bath for 20 min, then centrifuge to separate the phases. Decant the clear supernatant into a suitable stoppered flask, and allow to warm to room temperature. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for beclomethasone dipropionate and betamethasone valerate are 1.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.5 between betamethasone valerate and beclomethasone dipropionate Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H29FO5 in the portion of Cream taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Betamethasone Valerate RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of betamethasone, 392.46 Mr1 = molecular weight of betamethasone valerate, Mr2 476.59 Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight containers. USP REFERENCE STANDARDS 11 USP Beclomethasone Dipropionate RS USP Betamethasone Valerate RS RU RS CS

Analysis Samples: Sample solution and Standard solution Allow the spots to dry, and develop the chromatogram in a solvent system, until the solvent front has moved threefourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. View the spots under UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:2) Diluent: Glacial acetic acid in methanol (1 in 1000) Internal standard solution: 2 mg/mL of beclomethasone dipropionate in chloroform Standard stock solution: 1.6 mg/mL of USP Betamethasone Valerate RS in chloroform Standard solution: Pipet 2 mL of Standard stock solution into a 50-mL centrifuge tube, add 10 mL of 0.1 N hydrochloric acid, then add 2.0 mL of Internal standard solution. Insert the stopper into the tube, shake vigorously for 2 min, and centrifuge to separate the phases. Using a syringe, transfer the lower chloroform phase to a small stoppered vial. Evaporate the chloroform on a steam bath, at low heat, with the aid of a stream of nitrogen. Add 4.0 mL of Diluent, and swirl to dissolve the residue. Sample solution: Equivalent to 2.5 mg of betamethasone from Lotion in a stoppered, 50-mL centrifuge tube. Add 10.0 mL of 0.1 N hydrochloric acid, insert the stopper, and shake to disperse the specimen. Add 2.0 mL of chloroform and 2.0 mL of Internal standard solution. Insert the stopper into the tube, shake vigorously for 2 min, and centrifuge to separate the phases. Using a syringe, transfer the lower chloroform phase to a small stoppered vial. Evaporate the chloroform on a steam bath, at low heat, with the aid of a stream of nitrogen. Add 4.0 mL of Diluent, and swirl to dissolve the residue. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone valerate and beclomethasone dipropionate are 1.0 and 1.7, respectively.] Suitability requirements Resolution: NLT 4.5 between betamethasone valerate and beclomethasone dipropionate Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H29FO5 in the portion of Lotion taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 RU RS CS CU Mr1 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Betamethasone Valerate RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL) = molecular weight of betamethasone, 392.46
Betamethasone Valerate Lotion

(Comment on this Monograph)id=m9210=Betamethasone Valerate Lotion=B-Monos.pdf) DEFINITION Betamethasone Valerate Lotion contains an amount of Betamethasone Valerate (C27H37FO6) equivalent to NLT 95.0% and NMT 115.0% of the labeled amount of betamethasone (C22H29FO5). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 0.6 mg/mL of USP Betamethasone Valerate RS in a mixture of methanol and chloroform (2:1) Sample solution: Equivalent to 0.5 mg/mL of betamethasone from Lotion, in a mixture of methanol and chloroform (2:1) Application volume: 20 L Developing solvent system: Chloroform and ethyl acetate (1:1)
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the phases. Decant the clear supernatant into a suitable stoppered flask, and allow to warm to room temperature. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone valerate and beclomethasone dipropionate are 1.0 and 1.7, respectively.] Suitability requirements Resolution: NLT 4.5 between betamethasone valerate and beclomethasone dipropionate Relative standard deviation: NMT 2.0% Analysis Sample: Sample solution and Standard solution Calculate the percentage of C22H29FO5 in the portion of Ointment taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Betamethasone Valerate RS in the Standard solution (mg/mL) = nominal concentration of betamethasone in the CU Sample solution (mg/mL) Mr1 = molecular weight of betamethasone, 392.46 Mr2 = molecular weight of betamethasone valerate, 476.59 Acceptance criteria: 90.0%110.0% RU RS CS SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Betamethasone Valerate RS
= molecular weight of betamethasone valerate, 476.59 Acceptance criteria: 95.0%115.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa MINIMUM FILL 755: Meets the requirements PH 791: 4.06.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Betamethasone Valerate RS
Betamethasone Valerate Ointment

(Comment on this Monograph)id=m9220=Betamethasone Valerate Ointment=B-Monos.pdf) DEFINITION Betamethasone Valerate Ointment contains an amount of betamethasone valerate (C27H37FO6) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5), in a suitable ointment base. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Transfer the equivalent to 2 mg of betamethasone from Ointment to a separator. Add 20 mL of water and 2 mL of dilute hydrochloric acid (1 in 120). Extract with four 50-mL portions of chloroform, and combine the extracts. Filter through a cotton pledget, previously layered over with anhydrous sodium sulfate. Evaporate the filtrates on a steam bath under a stream of dry nitrogen to dryness. Dissolve the residue in alcohol to obtain a solution containing 1 mg/mL. Developing solvent system: Toluene and ethyl acetate (1:1) Application volume: 10 L Analysis Samples: Sample solution and Standard solution Proceed as directed in the chapter. Spray the plate with a mixture of sulfuric acid, methanol, and nitric acid (10:10:1), and heat at 105 for 15 min. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:2) Diluent: Glacial acetic acid in methanol (1 in 1000) Internal standard solution: 0.4 mg/mL of beclomethasone dipropionate in Diluent Standard stock solution: 0.6 mg/mL of USP Betamethasone Valerate RS in Diluent Standard solution: Standard stock solution and Internal standard solution (1:2) [NOTEThis solution has a known concentration of 0.2 mg/mL of USP Betamethasone Valerate RS.] Sample solution: Equivalent to 2.5 mg of betamethasone from Ointment, in a 50-mL centrifuge tube. Add 10.0 mL of the Internal standard solution and 5.0 mL of a solution of glacial acetic acid in alcohol (1 in 1000). Insert the stopper into the tube, and place in a water bath held at 70 until the specimen melts. Remove from the bath, and shake vigorously until the specimen resolidifies. Repeat the heating and shaking two more times. Place the tube in an icemethanol bath for 20 min, then centrifuge to separate
Betaxolol Hydrochloride
(Comment on this Monograph)id=m9230=Betaxolol Hydrochloride=B-Monos.pdf)
343.89 C18H29NO3 HCl 2-Propanol, 1-[4-[2-(cyclopropylmethoxy)ethyl]phenoxy]-3-(1methylethyl)amino-, hydrochloride, ()-; ()-1-[p-[2-(Cyclopropylmethoxy) ethyl]phenoxy]-3(isopropylamino)-2-propanol hydrochloride. [63659-18-7]. DEFINITION Betaxolol Hydrochloride contains NLT 98.5% and NMT 101.5% of C18H29NO3 HCl, calculated on the dried basis.
USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Chloride 191: It meets the requirements of the test, when tested as directed for alkaloidal hydrochlorides. ASSAY PROCEDURE Sample: 300 mg of Betaxolol Hydrochloride Analysis: Dissolve in 50 mL of glacial acetic acid. Add 7 mL of mercuric acetate TS and titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 34.39 mg of C18H29NO3 HCl. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Solution A: 0.025 M monobasic potassium phosphate containing 0.1% (w/v) of tetrabutylammonium bromide. Adjust with 0.025 M phosphoric acid to a pH of 3.0. Mobile phase: Acetonitrile and Solution A (3:17) System suitability solution: 2 mg/mL of USP Betaxolol Hydrochloride RS and 1 mg/mL of alprenolol hydrochloride in Mobile phase Sample solution: 2.0 mg/mL of Betaxolol Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 273 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for alprenolol and betaxolol are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.0 between the two peaks Tailing factor: NMT 2.0 for the two peaks Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution [NOTEAllow the elution to continue for five times the elution time of the betaxolol peak before making the next injection.] Calculate the percentage of each impurity in the portion of Betaxolol Hydrochloride taken: Result = (rU/rT) 100 = area of each individual peak, other than the main betaxolol peak of the Sample solution rT = sum of the areas of all the peaks of the Sample solution Acceptance criteria: Sum of all impurities, NMT 1.0% rU SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 113117 PH 791: 4.56.5, in a solution (1 in 50) LOSS ON DRYING 731: Dry in vacuum at 65 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Betaxolol Hydrochloride RS
Official Monographs / Betaxolol 71
Betaxolol Ophthalmic Solution

(Comment on this Monograph)id=m9235=Betaxolol Ophthalmic Solution=B-Monos.pdf) DEFINITION Betaxolol Ophthalmic Solution is a sterile, aqueous, isotonic solution of Betaxolol Hydrochloride. It contains a suitable antimicrobial preservative. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of betaxolol (C18H29NO3). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 2.75 mg/mL of USP Betaxolol Hydrochloride RS in water Sample solution: Equivalent to 2.5 mg/mL of betaxolol from a portion of Ophthalmic Solution diluted with water Mode: TLC Adsorbent: 0.25-mm layer of silica gel Application volume: 5 L Developing solvent system: Chloroform, isopropyl alcohol, and ammonium hydroxide (70:30:2) Spray reagent: A solution (1 in 1000) of ninhydrin in isopropyl alcohol Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram in a chromatographic chamber, using the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Spray the plate with Spray reagent and heat the plate at 105 for 10 min. Locate the spots on the plate. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Solution A: 7.1 g/L of anhydrous dibasic sodium phosphate in water, adjust with phosphoric acid to a pH of 3.0 Mobile phase: Acetonitrile and Solution A (1:1) Standard solution: 0.11 mg/mL of USP Betaxolol Hydrochloride RS in Solution A Sample solution: Nominally 0.1 mg/mL. Transfer a volume of Ophthalmic Solution, equivalent to about 10 mg of betaxolol, to a 100-mL volumetric flask. Dilute with Solution A to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: For the main betaxolol peak, between 1 and 3 Column efficiency: NLT 750 theoretical plates Tailing factor: 0.82.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H29NO3 in each mL of the Ophthalmic Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
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Betaxolol / Official Monographs

CS rU rS CS
USP 32
= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Betaxolol Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of betaxolol hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 274 nm Sample solutions: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Standard solution: USP Betaxolol Hydrochloride RS in Medium [NOTEA 5-cm pathlength cell may be used for lower dosage levels.] Tolerances: NLT 80% (Q) of the labeled amount of C18H29NO3 HCl UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Standard solution: 0.1 mg/mL of USP Betaxolol Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: Place 1 Tablet in a volumetric flask of appropriate size to obtain a concentration of 0.1 mg/mL based on the labeled claim. Add an amount of 0.1 N hydrochloric acid equal to 70% of the volume of the flask, shake by mechanical means until dissolved, dilute with 0.1 N hydrochloric acid to volume, and mix. Filter the mixture, discarding the first 20 mL of the filtrate. Blank: 0.1 N hydrochloric acid Mode: Spectrophotometry Analytical wavelength: 274 nm Cell: 1 cm Analysis Samples: Blank, Standard solution, and Sample solution Calculate the percentage of C18H29NO3 HCl in the Tablet taken: (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Betaxolol Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of betaxolol hydrochloride in the Sample solution (mg/mL)
= concentration of USP Betaxolol Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of betaxolol, 307.43 Mr1 = molecular weight of betaxolol hydrochloride, Mr2 343.89 Acceptance criteria: 90.0%110.0% SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791: 4.08.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Betaxolol Hydrochloride RS
Betaxolol Tablets
(Comment on this Monograph)id=m9238=Betaxolol Tablets=BMonos.pdf) DEFINITION Betaxolol Tablets contain an amount of Betaxolol Hydrochloride equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betaxolol hydrochloride (C18H29NO3 HCl). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Acetonitrile and water (1:1) Mobile phase: Acetonitrile, methanol, and 0.025 M pH 6.0 ammonium phosphate buffer (7:6:7) Mix, and degas under vacuum while stirring. Standard solution: 2 mg/mL of USP Betaxolol Hydrochloride RS in Diluent Sample solution: Dissolve NLT 20 Tablets in an appropriate, accurately measured volume of Diluent so that the final concentration is nominally 2 mg/mL of betaxolol hydrochloride. Sonicate until the Tablets are disintegrated. Cool to room temperature, dilute with Diluent to volume, and filter. Use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 273 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 3.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H29NO3 HCl in each Tablet: Result = (rU/rS) (CS/CU) 100
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Tablets to state both the content of the betaxolol active moiety and the content of betaxolol hydrochloride used in formulating them. USP REFERENCE STANDARDS 11 USP Betaxolol Hydrochloride RS
USP 32
Official Monographs / Bethanechol 73

Suitability requirements Resolution: NLT 0.8 between 2-hydroxypropyltrimethyl ammonium chloride and bethanechol, System suitability solution Tailing factor: NMT 3.5, Standard solution Relative standard deviation: NMT 3.0%, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of C7H17ClN2O2 in the portion of Bethanechol Chloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bethanechol Chloride RS in the Standard solution (mg/mL) = concentration of Bethanechol Chloride in the CU Sample solution (mg/mL) Acceptance criteria: 98.0%101.5% rU rS CS OTHER COMPONENTS CONTENT OF CHLORIDE Sample: 400 mg, previously dried Analysis: Dissolve in 30 mL of water, and add 40.0 mL of 0.1 N silver nitrate VS, then add 3 mL of nitric acid and 5 mL of nitrobenzene. Shake for a few min, add 2 mL of ferric ammonium sulfate TS, and titrate the excess silver nitrate with 0.1 N ammonium thiocyanate VS. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Cl. Acceptance criteria: 17.7%18.3% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method I 231: NMT 30 ppm Sample solution: 667 mg in 10 mL, add 2 mL of 1 N acetic acid, and dilute to 25 mL Organic Impurities PROCEDURE Solution A: 0.48 mg/mL of methanesulfonic acid Mobile phase, System suitability solution, and Sample solution: Proceed as directed in the Assay. Standard solution: 1 g/mL of USP Bethanechol Chloride RS in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode, Detector, Column, Temperature, and Flow rate: Proceed as directed in the Assay. Injection size: 50 L System suitability: Proceed as directed in the Assay. Suitability requirements Resolution: Proceed as directed in the Assay. Relative standard deviation: NMT 10.0% for bethanechol chloride in the Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of each impurity in the portion of Bethanechol Chloride: Result = (rU/rS) (CS/CU) F 100 rU rS CS = peak response for any impurity in the Sample solution = peak response of USP Bethanechol Chloride RS in the Standard solution = concentration of USP Bethanechol Chloride RS in the Standard solution (mg/mL)
Bethanechol Chloride
(Comment on this Monograph)id=m9350=Bethanechol Chloride=B-Monos.pdf)
196.67 C7H17ClN2O2 1-Propanaminium, 2-(aminocarbonyl)oxy-N,N,N-trimethyl-, chloride, ()-; ()-(2-Hydroxypropyl)trimethylammonium chloride carbamate [590-63-6]. DEFINITION Bethanechol Chloride contains NLT 98.0% and NMT 101.5% of C7H17ClN2O2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. PROCEDURE Sample solution: 50 mg in 2 mL Analysis: To the Sample solution, add 0.1 mL of cobaltous chloride solution (1 in 100), then add 0.1 mL of potassium ferrocyanide TS. Acceptance criteria: An emerald-green color is produced, and almost entirely fades in 510 min (as distinguished from choline chloride, which gives the same reaction but the color does not fade). C. PROCEDURE Sample solution: 10 mg/mL Analysis: To 1 mL of a solution, add 0.1 mL of iodine TS. Acceptance criteria: A brown precipitate is formed, and it rapidly changes to a dark olive-green color. D. IDENTIFICATION TESTSGENERAL, Chloride 191 ASSAY PROCEDURE Solution A: Transfer 29 mg of edetic acid to a 1000-mL volumetric flask, and dissolve in 500 mL. Add 300 L of nitric acid to the volumetric flask, and dilute to volume. Pass through a 0.45-m nylon membrane filter. Mobile phase: Acetonitrile and Solution A (1:19) System suitability solution: 25 mg of Bethanechol Chloride in 10 mL of 0.1 N sodium hydroxide in a 250-mL volumetric flask. Allow to stand for 15 min. Add 10 mL of 0.1 N hydrochloric acid. Dissolve in and dilute with Mobile phase to volume. Standard solution: 0.1 mg/mL of USP Bethanechol Chloride RS in Mobile phase Sample solution: 0.1 mg/mL of Bethanechol Chloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Conductivity Column: 3.9- 150-mm; packing L55 Temperature Detector: 35 Column: 30 Flow rate: 1 mL/min Injection size: 25 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for 2hydroxypropyltrimethyl ammonium chloride and bethanechol are 0.9 and 1.0, respectively.]
74
Bethanechol / Official Monographs

= concentration of Bethanechol Chloride taken to prepare the Sample solution (mg/mL) F = relative response factor, 0.79 for 2hydroxypropyltrimethyl ammonium and 1.0 for any other impurity Acceptance criteria Individual impurities: NMT 1.0% of 2hydroxypropyltrimethyl ammonium is found; NMT 0.1% of any other impurity is found. Total impurities: NMT 1.5% CU
USP 32
Mode: LC Detector: Conductivity Column: 4-mm 25-cm; packing L53 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: System suitability solution [NOTEThe relative retention times for sodium, magnesium, calcium, 2-hydroxypropyltrimethyl ammonium chloride, and bethanechol are 1.0, 1.4, 1.6, 2.0, and 2.8, respectively. ] Suitability requirements Resolution: NLT 2.0, between the calcium ion and 2hydroxypropyltrimethyl ammonium chloride Column efficiency: NLT 350 theoretical plates, determined from the bethanechol peak Tailing factor: NMT 4.5 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C7H17ClN2O2 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bethanechol Chloride RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 95.0%105.0% rU rS CS IMPURITIES Organic Impurities PROCEDURE: LIMIT OF 2-HYDROXYPROPYLTRIMETHYL AMMONIUM CHLORIDE Diluent, Mobile phase, System suitability solution, and Chromatographic system: Prepare as directed in the Assay. 2-Hydroxypropyltrimethyl ammonium chloride solution: Transfer 50 mg of bethanechol chloride to a 50-mL volumetric flask. Add 40 mL of 0.1 N sodium hydroxide, and sonicate until fully dissolved. Dilute with 0.1 N sodium hydroxide to volume, and allow to stand for 5 days to allow adequate time for conversion from bethanechol to 2hydroxypropyltrimethyl ammonium chloride. Chromatograph as directed in Analysis to verify the presence and location of the peak for 2hydroxypropyltrimethyl ammonium chloride. Standard solution: Use the Standard solution, prepared as directed in the Assay. Sample solution: Use the Sample solution, prepared as directed in the Assay. System suitability: Proceed as directed in the Assay. Analysis Samples: 2-Hydroxypropyltrimethyl ammonium chloride solution, Sample solution, and Standard solution Calculate the percentage of 2-hydroxypropyltrimethyl ammonium chloride in each mL of Injection taken: Result = (rU/rS) (CS/CU) 100 rU = peak response for 2-hydroxypropyltrimethyl ammonium chloride from the Sample solution
SPECIFIC TESTS PH 791: 5.56.5, in a solution (1 in 100) LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Bethanechol Chloride RS
Bethanechol Chloride Injection

(Comment on this Monograph)id=m9400=Bethanechol Chloride Injection=B-Monos.pdf) DEFINITION Bethanechol Chloride Injection is a sterile solution of Bethanechol Chloride in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of C7H17ClN2O2. IDENTIFICATION A. PROCEDURE Sample solution: 25 mg/mL of bethanechol chloride from the Injection Analysis: To the Sample solution, add 0.1 mL of cobaltous chloride solution (1 in 100), then add 0.1 mL of potassium ferrocyanide TS. Acceptance criteria : An emerald-green color is produced, and almost entirely fades in 5 to 10 min (distinction from choline chloride, which gives the same reaction but the color does not fade). B. PROCEDURE Sample solution : 10 mg/mL from the Injection Analysis: To 1 mL of Sample solution add 0.1 mL of iodine TS. Acceptance criteria: A brown precipitate is formed, and it rapidly changes to a dark olive-green color. C. IDENTIFICATION TESTSGENERAL, Chloride 191 ASSAY PROCEDURE Diluent: 0.1 mg/mL of calcium chloride and 0.1 mg/mL of magnesium chloride Mobile phase: 20 mM methanesulfonic acid System suitability solution: To 25 mg of USP Bethanechol Chloride RS add 15 mL of water, 2 mL of Diluent, and 0.5 mL of 0.1 N of sodium hydroxide. Dilute with water to 25 mL. Standard solution: 1 mg/mL of USP Bethanechol Chloride RS Sample solution: 1 mg/mL of Bethanechol Chloride from a volume of Injection Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
= peak response for bethanechol from the Standard solution = concentration of USP Bethanechol Chloride RS CS in the Standard solution (mg/mL) = concentration of bethanechol chloride in the CU Sample solution (mg/mL) Acceptance criteria: NMT 4.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 25.0 USP Endotoxin Units/mg of bethanechol chloride PH 791: 5.57.5 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Bethanechol Chloride RS USP Endotoxin RS rS
Official Monographs / Bethanechol 75

Suitability requirements Relative standard deviation: NMT 3.1% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H17ClN2O2 in the volume of Oral Solution: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bethanechol Chloride RS in the Standard solution (g/mL) = nominal concentration of bethanechol chloride CU in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 3.94.9 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at room temperature or in a cold place. LABELING: Label it to state the beyond-use date. BEYOND-USE DATE: 60 days after the day on which it was compounded. USP REFERENCE STANDARDS 11 USP Bethanechol Chloride RS rU rS CS
Bethanechol Chloride Oral Solution

(Comment on this Monograph)id=m1598=Bethanechol Chloride Oral Solution=B-Monos.pdf) DEFINITION Bethanechol Chloride Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of bethanechol chloride (C7H17ClN2O2). Prepare Bethanechol Chloride Oral Solution 5 mg/mL as follows (See Pharmaceutical CompoundingNonsterile Preparations 795).
Bethanechol Chloride Vehicle for Oral Solution (regular or sugarfree), NF To make 500 mg A sufficient quantity 100 mL
Bethanechol Chloride Oral Suspension

(Comment on this Monograph)id=m1381=Bethanechol Chloride Oral Suspension=B-Monos.pdf) DEFINITION Bethanechol Chloride Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of bethanechol chloride (C7H17ClN2O2). Prepare Bethanechol Chloride Oral Suspension 5 mg/mL as follows (See Pharmaceutical CompoundingNonsterile Preparations 795).
Bethanechol Chloride Vehicle: a mixture of Vehicle for Oral Solution, (regular or sugar-free), NF and Vehicle for Oral Suspension, NF (1:1) To make 500 mg A sufficient quantity 100 mL
Add Bethanechol Chloride powder and about 20 mL of Vehicle to a mortar, and mix. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add enough Vehicle to bring to final volume, and mix well. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (33:67) Standard solution: 500 g/mL of USP Bethanechol Chloride RS in Mobile phase Sample solution: Agitate the container of Oral Solution for 30 min on a rotating mixer, remove a 10-mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the sample from the freezer, allow it to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 2.0 mL of the sample into a 20-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 200 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 0.7 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTERetention time: about 3 min]
If using Bethanechol Chloride Tablets, add to a suitable mortar and comminute to a fine powder, or add the Bethanechol Chloride powder to the mortar. Add about 20 mL of the Vehicle, and mix to a uniform paste. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add sufficient Vehicle to final volume, and mix well. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (33:67) Standard solution: 500 g/mL of USP Bethanechol Chloride RS in Mobile phase Sample solution: Agitate the container of Oral Suspension for 30 min on a rotating mixer, remove a 10-mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the sample from the freezer, allow it to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 5.0 mL of the sample into a 50-mL volumetric flask, and dilute with Mobile phase to volume.
76
Bethanechol / Official Monographs
USP 32
0.1 N hydrochloric acid. Dissolve in and dilute with Mobile phase to volume. Standard solution: 0.1 mg/mL of USP Bethanechol Chloride RS in Mobile phase Sample solution: Equivalent to 0.1 mg/mL of bethanechol chloride from powdered Tablets (NLT 20 Tablets) in Mobile phase [NOTEIn a suitable volumetric flask, transfer a portion of the powder equivalent to 1 Tablet. Add an amount of Mobile phase, 60% to 70% of the total volume of the flask. Sonicate for 20 min. Shake by mechanical means for 15 min. Dilute with Mobile phase to volume, and mix. Allow to stand for 10 min, and pass the solution through a 1-m glass filter, discarding the first 3 mL of the filtrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Conductivity Column: 3.9- 150-mm; packing L55 Temperature Detector: 35 Column: 30 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for 2hydroxypropyltrimethyl ammonium chloride and bethanechol are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 0.8 between 2-hydroxypropyltrimethyl ammonium chloride and bethanechol, System suitabillity solution Tailing factor: NMT 3.5, Standard solution Relative standard deviation: NMT 3.0%, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of C7H17ClN2O2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bethanechol Chloride RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of C7H17ClN2O2 dissolved using the following method. Solution A, Mobile phase, and Chromatographic system: Proceed as directed in the Assay. Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Standard solution: USP Bethanechol Chloride RS in Medium System suitability: Proceed as directed in the Assay. Analysis Samples: Sample solution and Standard solution Calculate the quantity of C7H17ClN2O2 dissolved based on the peak responses obtained from the Sample solution and the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C7H17ClN2O2 rU rS CS
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 200 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 0.7 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTERetention time: about 3 min] Suitability requirements Relative standard deviation: NMT 3.1% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H17ClN2O2 in the volume of Oral Suspension: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bethanechol Chloride RS in the Standard solution (g/mL) = nominal concentration of bethanechol chloride CU in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 3.94.9 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at room temperature, or in a cold place. LABELING: Label it to state that it is to be well shaken, and to state the beyond-use date. Beyond-Use Date: 60 days after the day on which it was compounded. USP REFERENCE STANDARDS 11 USP Bethanechol Chloride RS
Bethanechol Chloride Tablets

(Comment on this Monograph)id=m9450=Bethanechol Chloride Tablets=B-Monos.pdf) DEFINITION Bethanechol Chloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of bethanechol chloride (C7H17ClN2O2). IDENTIFICATION INFRARED ABSORPTION 197M Sample solution: Equivalent to 100 mg of bethanechol chloride from pulverized Tablets. Add 15 mL of ether, and allow to digest for 15 min. Decant the ether, again extract the residue with 10 mL of ether, and discard the ether extracts. To the residue add 30 mL of alcohol, shake for 10 min, and allow to stand for 1 h with frequent agitation. Filter with suction, and evaporate the filtrate on a steam bath to dryness: the bethanechol chloride so obtained is recrystallized from alcohol and dried at 105 for 2 h. ASSAY PROCEDURE Solution A: Transfer 29 mg of edetic acid to a 1000-mL volumetric flask, and dissolve in 500 mL. Add 300 L of nitric acid to the volumetric flask, and dilute to volume. Pass through a 0.45-m nylon membrane filter. Mobile phase: Acetonitrile and Solution A (1:19) System suitability solution: Transfer 25 mg of bethanechol chloride in 10 mL of 0.1 N sodium hydroxide to a 250-mL volumetric flask. Allow to stand for 15 min. Add 10 mL of
USP 32
Official Monographs / Bicalutamide 77

Total impurities: NMT 1.5%
IMPURITIES Organic Impurities PROCEDURE Solution A: 0.48 mg/mL of methanesulfonic acid Mobile phase: Acetonitrile and Solution A (1:19) System suitability solution: Transfer 25 mg of bethanechol chloride in 10 mL of 0.1 N sodium hydroxide to a 250 mL volumetric flask. Allow to stand for 15 min. Add 10 mL of 0.1 N hydrochloric acid. Dissolve in and dilute with Mobile phase to volume. Standard solution: 1 g/mL of USP Bethanechol Chloride RS in Mobile phase Sample solution: Equivalent to 0.1 mg/mL of bethanechol chloride from powdered Tablets (NLT 20 Tablets) in Mobile phase [NOTEIn a suitable volumetric flask, transfer a portion of the powder equivalent to 1 Tablet. Add an amount of Mobile phase, 60% to 70% of the total volume of the flask. Sonicate for 20 min. Shake by mechanical means for 15 min. Dilute with Mobile phase to volume, and mix. Allow to stand for 10 min, and pass the solution through a 1-m glass filter, discarding the first 3 mL of the filtrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Conductivity Column: 3.9- 150-mm; packing L55 Temperature Detector: 35 Column: 30 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for 2hydroxypropyltrimethyl ammonium chloride and bethanechol are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 0.8 between 2-hydroxypropyltrimethyl ammonium chloride and bethanechol, System suitability solution Relative standard deviation: NMT 10.0% for bethanechol chloride, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of each impurity in the portion of Tablets taken: Result = (ru/rS) (CS/CU) F 100 = peak response for any impurity in the Sample solution rS = peak response of USP Bethanechol Chloride RS in the Standard solution CS = concentration of USP Bethanechol Chloride RS in the Standard solution (mg/mL) CU = concentration of bethanechol chloride in the portion taken for the Sample solution as determined in the Assay (mg/mL) F = relative response factor equal to 0.79 for 2hydroxypropyltrimethyl ammonium and 1.0 for any other impurity Acceptance criteria Individual impurities: NMT 1.0% of 2hydroxypropyltrimethyl ammonium chloride is found; NMT 0.2% of any other impurity is found ru
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Bethanechol Chloride RS
Bicalutamide
(Comment on this Monograph)id=m2641=Bicalutamide=BMonos.pdf)
430.37 C18H14F4N2O4S Propanamide, N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(4fluorophenyl)sulfonyl]-2-hydroxy-2-methyl-,(+)(+)-4-Cyano-,,-trifluoro-3-[(p-fluorophenyl) sulfonyl]-2-methyl-m-lactotoluidide [90357-06-5]. DEFINITION Bicalutamide contains NLT 98.0% and NMT 102.0% of C18H14F4N2O4S, calculated on the anhydrous and solvent-free basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, tetrahydrofuran, and water (6:3:11) Standard solution: 0.05 mg/mL of USP Bicalutamide RS in a minimum amount of tetrahydrofuran, and diluted with Mobile phase Sample solution: 0.05 mg/mL of Bicalutamide in a minimum amount of tetrahydrofuran, and diluted with Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 5-mm 25-cm; 5-m packing L1 Temperature: 3540 Flow rate: 1.8 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C18H14F4N2O4S in the portion of Bicalutamide taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
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CS
Impurity Table (continued) Relative Retention Time
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= concentration of USP Bicalutamide RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Mobile phase: Proceed as directed in the Assay. System suitability solution: 0.05 mg/mL of USP Bicalutamide RS and 0.02 mg/mL of USP Bicalutamide Related Compound B RS in a minimum amount of tetrahydrofuran, and dilute with Mobile phase Standard solution: 0.02 mg/mL of USP Bicalutamide RS in a minimum amount of tetrahydrofuran, and dilute with Mobile phase Sample solution: 4.0 mg/mL of Bicalutamide in a minimum amount of tetrahydrofuran, and dilute with Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 5-mm 25-cm; 5-m packing L1 Temperature: 3540 Flow rate: 1.8 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 2.0 between bicalutamide and bicalutamide related compound B Tailing factor: NMT 1.3 for bicalutamide Relative standard deviation: NMT 4.0% for bicalutamide Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Bicalutamide taken: Result = (rU/rS) (CS/CU) 100 = peak response for each impurity from the Sample solution rS = peak response for bicalutamide from the Standard solution CS = concentration of USP Bicalutamide RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria Individual impurities: See Impurity Table. Total impurities: NMT 0.5%
Impurity Table Relative Retention Time 0.74 0.78 1.19 Acceptance Criteria, NMT (%) 0.2 0.2 0.2
Name Individual unspecified impurity Total unspecified impurities

1
Acceptance Criteria, NMT (%) 0.1 0.3
(RS)-4-cyano-3-phenylsulfonyl-2-hydroxy-2-methyl-3-(trifluoromethyl)propionanilide. 2(RS)-4-cyano-3-(2-fluorophenylsulfonyl)-2-hydroxy-2-methyl-3(trifluoromethyl)-propionanilide. 3(RS)-4-cyano-3-(4-fluorophenylsulfonyl)-2-methyl-3(trifluoromethyl)propionanilide.
SPECIFIC TESTS WATER, Method I 921:
NMT 0.2%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at room temperature. USP REFERENCE STANDARDS 11 USP Bicalutamide RS USP Bicalutamide Related Compound B RS
Add the following:

v
Bicalutamide Tablets
(Comment on this Monograph)id=m9465=Bicalutamide Tablets=B-Monos.pdf) DEFINITION Bicalutamide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of bicalutamide (C18H14F4N2O4S). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Tetrahydrofuran, acetonitrile, and water (20:15:65). System suitability stock solution: 0.8 mg/mL of USP Bicalutamide RS and 0.4 mg/mL of USP Bicalutamide Related Compound B RS in tetrahydrofuran System suitability solution: 0.04 mg/mL of USP Bicalutamide RS and 0.02 mg/mL of USP Bicalutamide Related Compound B RS, from System suitability stock solution, in Mobile phase Standard stock solution: 0.8 mg/mL of USP Bicalutamide RS in tetrahydrofuran Standard solution: 0.04 mg/mL of USP Bicalutamide RS, from Standard stock solution, in Mobile phase Sample stock solution: Equivalent to 50 mg of bicalutamide, from powdered Tablets, in 50 mL of tetrahydrofuran. Sonicate for NLT 10 min to complete dissolution, and allow to cool. Dilute with tetrahydrofuran to 100.0 mL, and pass through a suitable filter having a porosity of 0.45 m. Sample solution: 0.04 mg/mL of bicalutamide, from Sample stock solution, in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
rU
Name Des-fluoro analog1 2-Fluoro isomer2 Des hydroxy analog3

1
(RS)-4-cyano-3-phenylsulfonyl-2-hydroxy-2-methyl-3-(trifluoromethyl)propionanilide. 2(RS)-4-cyano-3-(2-fluorophenylsulfonyl)-2-hydroxy-2-methyl-3(trifluoromethyl)-propionanilide. 3(RS)-4-cyano-3-(4-fluorophenylsulfonyl)-2-methyl-3(trifluoromethyl)propionanilide.
USP 32
Mode: LC Detector: UV 270 nm Column: 5-mm 12.5-cm; 3-m packing L1 Temperature: 50 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 1.9 between bicalutamide and bicalutamide related compound B Tailing factor: Less than 1.3 for bicalutamide Relative standard deviation: NMT 2.0% for bicalutamide Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H14F4N2O4S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU = peak area from the Sample solution = peak area from the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 1.0% (w/v) sodium lauryl sulfate in water; 1000 mL Apparatus 2: 50 rpm Time: 45 min Detector: UV 270 nm Standard solution: 0.05 mg/mL of USP Bicalutamide RS in Medium [NOTEDissolve the RS in a volume of tetrahydrofuran equal to 1% of the final solution volume before dilution with Medium.] Sample solution: Sample per Dissolution 711. Pass through a suitable filter having a porosity of 0.45 m. Blank: Medium Analysis Samples: Standard solution and Sample solution Determine the amount of C18H14F4N2O4S dissolved: Result = (AU/AS) (CS V)/L 100 = absorbance of the Sample solution AU = absorbance of the Standard solution AS = concentration of the Standard solution (mg/mL) CS V = volume of Medium (1000 mL) L = label claim (mg) Tolerances: NLT 80% (Q) of the labeled amount of C18H14F4N2O4S UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Spectrometric conditions Analytical wavelength: UV 270 nm Blank: Diluent Diluent: 10 mg/mL of sodium lauryl sulfate in water Standard solution: 0.05 mg/mL of USP Bicalutamide RS in Medium [NOTEDissolve the RS in a minimum volume of tetrahydrofuran before dilution with Diluent.] Sample solution: Transfer 1 Tablet to a 100-mL volumetric flask, add about 10 mL of water, and sonicate for approximately 30 min. Add about 80 mL of tetrahydrofuran, and sonicate for 30 min to complete dissolution of the bicalutamide. Allow to cool to room temperature, and dilute with tetrahydrofuran to volume. Pass a portion of the solution through a filter having a
Official Monographs / Bicalutamide 79

porosity of 0.45 m, and dilute 10.0 mL with Diluent to 100.0 mL. Analysis Samples: Standard solution, Sample solution, and Blank Calculate the percentage of C18H14F4N2O4S in each Tablet: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = = = = absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of the Sample solution (mg/mL)
IMPURITIES Organic Impurities PROCEDURE: LIMIT OF 4-AMINO-2-(TRIFLUOROMETHYL)BENZONITRILE Mobile phase and System suitability solution: Proceed as directed in the Assay. Standard stock solution: 0.2 mg/mL of USP Bicalutamide RS in tetrahydrofuran Standard solution: 0.02 mg/mL of USP Bicalutamide RS, from Standard stock solution, in Mobile phase Sample solution: Transfer an equivalent to 50 mg of bicalutamide, from powdered Tablets, to a 25-mL volumetric flask. Add 2.0 mL of tetrahydrofuran, and allow to stand for 5 min. Add 20 mL of Mobile phase, sonicate for NLT 10 min, and allow to cool. Dilute with Mobile phase to volume, and pass through a suitable filter having a porosity of 0.2 m. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 5-mm 12.5-cm; 3-m packing L1 Temperature: 50 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times of 4-amino-2(trifluoromethyl)benzonitrile and bicalutamide related compound B are about 0.4 and about 1.1, respectively.] Suitability requirements Resolution: NLT 1.9 between bicalutamide and bicalutamide related compound B Tailing factor: Less than 1.3 for bicalutamide Relative standard deviation: NMT 2.0% for bicalutamide Analysis Sample: Sample solution and Standard solution Calculate the percentage of 4-amino-2(trifluoromethyl)benzonitrile in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (1/RRF) x 100 = peak response of 4-amino-2(trifluoromethyl)benzonitrile from the Sample solution = peak response of bicalutamide from the rS Standard solution = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU RRF = relative response factor of 4-amino-2(trifluoromethyl)benzonitrile (1.4) Acceptance criteria: NMT 0.1% of 4-amino-2(trifluoromethyl)benzonitrile rU
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TOTAL VIABLE SPORE COUNT Analysis: Proceed as directed for Total Viable Spore Count. Acceptance criteria: The requirements of the test are met if the log average number of viable spores per carrier is NLT 0.3 log of the labeled spore count per carrier and does not exceed the log labeled spore count per carrier by 0.48. SPECIFIC TESTS PURITY Presence of contamination by other microorganisms: By examination of the spores on a suitable plate culture medium, there is no evidence of contamination with other microorganisms. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in the original package under the conditions recommended on the label, and protect from light, toxic substances, excessive heat, and moisture. The packaging and container materials do not adversely affect the performance of the article used as directed in the labeling. EXPIRATION DATE: The expiration date is determined on the basis of stability studies and is NLT 18 months from the date of manufacture. The date of manufacture is the date on which the first determination of the total viable spore count was made. LABELING: Label it to state that it is a Biological Indicator for Dry-Heat Sterilization, Paper Carrier; to indicate its D value and the method used to determine such D value, i.e., by spore count or fraction negative procedure after graded exposures to the sterilization conditions; the survival time and kill time under the specified sterilization conditions stated on the label; its particular total viable spore count, with a statement that such count has been determined after preliminary heat treatment; and its recommended storage conditions. State in the labeling the size of the paper carrier, the strain and ATCC number from which the spores were derived, and instructions for spore recovery and for safe disposal of the indicator. Indicate in the labeling that the stated D value is reproducible only under the exact conditions under which it was determined, that the user would not necessarily obtain the same result, and that the user should determine the suitability of the biological indicator for the particular use. DISPOSAL: Before destruction or discard, sterilize it by steam at 121 for NLT 30 min, or by NLT an equivalent method recommended by the manufacturer. This includes a test strip used in any test procedure for the strips themselves.
NMT 5.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Bicalutamide RS USP Bicalutamide Related Compound B RSvUSP32
Biological Indicator for Dry-Heat Sterilization, Paper Carrier

(Comment on this Monograph)id=m9497=Biological Indicator for Dry-Heat Sterilization, Paper Carrier=B-Monos.pdf) DEFINITION Biological Indicator for Dry-Heat Sterilization, Paper Carrier, is a defined solution of viable spores made from a culture derived from a specified strain of Bacillus subtilis subspecies niger, on a suitable grade of paper carrier, individually packaged in a container readily penetrable by dry heat, and characterized for predictable resistance to dry-heat sterilization. The packaged Biological Indicator for Dry-Heat Sterilization, Paper Carrier, has a particular labeled spore count per carrier of NLT 104 and NMT 109 spores. When labeled for and subjected to dry-heat sterilization conditions at a particular temperature, it has a survival time and kill time appropriate to the labeled spore count and to the decimal reduction value (D value, in min) of the solution, specified by: Survival time (in min) = NLT (labeled D value) (log labeled spore count per carrier 2); and Kill time (in min) = NMT (labeled D value) (log labeled spore count per carrier + 4) IDENTIFICATION The biological indicator organism complies substantially with the morphological, cultural, and biochemical characteristics of the strain of Bacillus subtilis, ATCC No. 9372, designated subspecies niger, detailed for that biological indicator organism under Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier. PERFORMANCE TESTS [NOTESee Biological IndicatorsResistance Performance Tests 55] D VALUE Analysis: Proceed as directed for D Value Determination. Acceptance criteria: The requirements of the test are met if the determined D value is within 20% of the labeled D value for the selected sterilizing temperature, and if the confidence limits of the estimate are within 10% of the determined D value. SURVIVAL TIME and KILL TIME Analysis: Proceed as directed for Survival Time and Kill Time. Acceptance criteria: The requirements of the test are met if all of the specimens subjected to dry-heat sterilization for the survival time show evidence of growth, while none of the specimens subjected to dry-heat sterilization for the kill time show growth. If for either the survival time test or the kill time test NMT one specimen out of both groups fails the survival requirement or the kill requirement (whichever is applicable), continue the corresponding test with four additional groups, each consisting of 20 specimens, according to the procedure described. If all of the additional specimens subjected to dry-heat sterilization either meet the survival requirement for the survival time test, or meet the kill requirement for the kill time test, whichever is applicable, the requirements of the test are met.
Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier

(Comment on this Monograph)id=m9500=Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier=B-Monos.pdf) DEFINITION Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier, is a defined solution of viable spores made from a culture derived from a specified strain of Bacillus subtilis subspecies niger on a suitable grade of paper carrier, individually packaged in a suitable container readily penetrable by ethylene oxide sterilizing gas mixture, and characterized for predictable resistance to sterilization with such gas mixture. The packaged Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier, has a particular labeled spore count per carrier of NLT 104 and NMT 109 spores. Where labeled for and subjected to particular ethylene oxide sterilization conditions of a stated gaseous mixture, temperature, and relative humidity, it has a survival time and kill time
USP 32
appropriate to the labeled spore count and to the decimal reduction value (D value, in min) of the solution, specified by: Survival time (in min) = NLT (labeled D value) (log labeled spore count per carrier 2), and Kill time (in min) = NMT (labeled D value) (log labeled spore count per carrier + 4). IDENTIFICATION The biological indicator organism complies substantially with the morphological, cultural, and biochemical characteristics of the strain of Bacillus subtilis, ATCC No. 9372, designated subspecies niger: under microscopic examination, it consists of Gram-positive rods of width 0.7 to 0.8 m, and length 2 to 3 m; the endospores are oval and central, and the cells are not swollen; when incubated aerobically in appropriate media at 3035, growth occurs within 24 h, and similar inoculated media incubated concomitantly at 5560 show no evidence of growth in the same period; agar colonies have a dull appearance and may be cream or browncolored; when incubated in nutrient broth, it develops a pellicle and shows little or no turbidity; when examined under conventional biochemical tests for microbial characterization, develops a black pigment with tyrosine, it liquefies gelatin, utilizes citrate but not propionate or hippurate, reduces nitrate, and hydrolyzes both starch and glucose with no gas production; it shows a positive catalase reaction and gives a positive result with the Voges-Proskauer test. PERFORMANCE TESTS [NOTESee Biological IndicatorsResistance Performance Tests 55.] D VALUE Analysis: Proceed as directed in the relevant procedure for D value. Acceptance criteria: The requirements of the test are met if the determined D value is within 20% of the labeled D value for the selected sterilizing temperature, and if the confidence limits of the estimate are within 10% of the determined D value. SURVIVAL TIME and KILL TIME Analysis: Proceed as directed for Survival Time and Kill Time, using the procedure for Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier. Acceptance criteria: The requirements of the test are met if all of the specimens subjected to the ethylene oxide sterilization conditions for the survival time show evidence of growth, while none of the specimens subjected to the ethylene oxide sterilization conditions for the kill time shows evidence of growth. If for either the survival time test or the kill time test, NMT 1 specimen out of both groups fails the survival requirement or the kill requirement (whichever is applicable), continue the corresponding test with 4 additional groups, each consisting of 20 specimens, according to the procedure described. If all the additional specimens subjected to ethylene oxide sterilization meet either the survival requirement for the Survival time test or the kill requirement for the Kill time test, whichever is applicable, the requirements of the test are met. TOTAL VIABLE SPORE COUNT Analysis: Proceed as directed for Total Viable Spore Count, using the procedure for Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier. Acceptance criteria: The requirements of the test are met if the log average number of viable spores per carrier is NLT 0.3 log of the labeled spore count per carrier and does not exceed the log labeled spore count per carrier by 0.48.
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SPECIFIC TESTS PURITY (Presence of contamination by other microorganisms): By examination of the spores on a suitable plate culture medium, there is no evidence of contamination with other microorganisms. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in the original package under the conditions recommended on the label, and protect it from light, toxic substances, excessive heat, and moisture. The packaging and container material shall be such that it does not adversely affect the performance of the article used as directed in the labeling. EXPIRATION DATE: The expiration date is determined on the basis of stability studies and is NLT 18 months from the date of manufacture, the date of manufacture being the date on which the first determination of the total viable spore count was made. LABELING: Label it to state that it is a Biological Indicator for Ethylene Oxide Sterilization, Paper Carrier; to indicate its D value, the method used to determine such D value, i.e., by spore count or fraction negative procedure after graded exposures to the sterilization conditions; the Survival time and Kill time under specified sterilization conditions stated on the label; its particular Total viable spore count, with a statement that such count has been determined after preliminary heat treatment; and its recommended storage conditions. State in the labeling the size of the paper carrier, the strain, and ATCC number from which the spores were derived, and instructions for spore recovery and for safe disposal of the indicator. Indicate in the labeling that the stated D value is reproducible only under the exact conditions under which it was determined, that the user would not necessarily obtain the same result, and that the user should determine the suitability of the biological indicator for the particular use. DISPOSAL: Prior to destruction or discard, sterilize it by steam at 121 for NLT 30 min, or by NLT an equivalent method recommended by the manufacturer. This includes a strip used in test procedures for strips themselves.
Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Liquid Spore Suspensions
(Comment on this Monograph)id=m825=Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Liquid Spore Suspensions=B-Monos.pdf) DEFINITION Liquid spore suspensions may be used to prepare biological indicators for moist heat, dry heat, and gaseous modes of sterilization. On the basis of the intended sterilization use, the suspension is prepared inoculated from a culture of viable spores derived from one of several sterilization resistant microorganisms. Cultures used for liquid spore suspensions include, among others, the following: Clostridium sporogenes, Geobacillus stearothermophilus (formerly B. stearothermophilus), Bacillus atrophaeus (formerly B. subtilis), Bacillus subtilis, or Bacillus coagulans. Each tube or container containing the spore suspension is individually packaged for use. The packaged biological indicator spore suspension has a particular labeled spore count of NLT 103, and NMT 109, spores/mL of suspension. The suspending medium or vehicle is identified according to chemical composition. It has a survival time and
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toxic substances, and excessive heat. The materials of composition of the tube or container must not adversely affect the performance of the spore suspension. EXPIRATION DATE: The expiration date is determined on the basis of stability studies. The date of manufacture is the date on which the first determination of the total viable count was made. SHIPMENT: Spore suspensions must be shipped following EPA requirements for the shipment of biological and/or etiological agents. DISPOSAL: Spore suspensions that a user or manufacturer wishes to dispose of are first sterilized by moist heat by a process that achieves temperatures of approximately 121 for NLT 30 min. Alternative sterilization methods yielding equivalent or greater levels of lethality may be used. LABELING: Label the spore suspension tube or container or package insert to state that it is a biological indicator spore suspension for use in label specified applications for moistheat, dry-heat, and/or gaseous sterilization. State the biological indicator D value obtained under defined exposures to stated sterilization conditions using the Survival Curve Method of D value analysis. State the Survival time and kill time for the biological indicator suspension under specified conditions on the label. The total viable spore count per mL of the suspension following heat shock treatment must also appear on the label. State in the labeling the strain and ATCC number of the microorganisms used in the spore suspension and instructions for spore recovery and for safe disposal of the suspension. Indicate in the labeling that the stated D value is reproducible only under the exact conditions determined by the manufacturer and that the user would not necessarily obtain the same results if different exposure conditions were used. State that the user should determine the suitability of the biological indicator spore suspension for the users particular purpose and exposure conditions.
kill time appropriate to the labeled spore count, and to the decimal reduction value (the D value, in min), specified by the following: Survival time (in min) = NLT (labeled D value) (log of labeled spore count per mL from 1:100 dilution of original suspension 2); and Kill time (in min) = NMT (labeled D value) (log of labeled spore count per mL from 1:100 dilution of original suspension + 4). IDENTIFICATION Identification for the biological indicator is of lesser importance than the more relevant concerns of population and resistance to the sterilization processes. The manufacturer should identify the species used. PERFORMANCE TESTS [NOTESee Biological IndicatorsResistance Performance Tests 55, Total Viable Spore Count] D VALUE Analysis: If the biological indicators are being used in moist-heat or dry-heat sterilization, proceed as directed in the relevant procedure in D Value Determination. Acceptance criteria: The requirements of the test are met if the determined D value is within 20% of the labeled D value for the selected sterilizing conditions, and if the confidence limits of the estimate are within 10% of the determined D value. The D value determination method used should be that identified by the biological indicator manufacturer. SURVIVAL TIME and KILL TIME Analysis: Follow the procedure under Survival Time and Kill Time in D Value Determination. The test is conducted using 1:100 dilution aliquots of the original suspension to inoculate carrier substrates that are most likely to be used by the purchaser of the spore suspensions for a given mode of sterilization. Following a total viable count analysis, the inoculated substrates are subjected to sterilization exposure conditions intended to indicate survival. Acceptance criteria: The inoculated carriers must show evidence of growth among the exposed carriers. Analysis: A second study is conducted to demonstrate the conditions necessary to result in total kill of the carriers. Acceptance criteria: None of the carriers subjected to conditions designed to induce total kill should show growth. If for either the survival-time test or the kill-time test, NMT one carrier out of both groups fails the survival or kill requirements, continue the corresponding test with four additional groups, each consisting of 10 carriers, according to the procedure described. For biological indicators for use with moist-heat or dry-heat sterilization, if all of the additional specimens subjected to the specific sterilization process either meet the survival requirements for the survival-time test or meet the kill requirement for the killtime test, whichever is applicable, the requirements are met. TOTAL VIABLE SPORE COUNT: Analysis: Proceed as directed for Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Liquid Spore Suspensions in Total Viable Spore Count Acceptance criteria The requirements for this test are met if the total viable spore count within the suspension is within 1 log of the value stipulated by the manufacturer. 103 - 109 spores/mL of suspension. SPECIFIC TESTS PURITY: There is no evidence of contamination with other microorganisms following examination of spores recovered from the metal carriers using suitable plate-culture medium. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in the original tube or container under the conditions recommended on the label, and protect the contents of the tube or container from light,
Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Nonpaper Carriers
(Comment on this Monograph)id=m663=Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Nonpaper Carriers=B-Monos.pdf) DEFINITION Biological indicators for moist heat, dry heat, and gaseous modes of sterilization may be nonpaper carriers inoculated with a culture of viable spores derived from one of several sterilization-resistant microorganisms, based on the intended sterilization use. Cultures used for inoculation of carriers include, among others, Clostridium sporogenes, Geobacillus stearothermophilus (formerly B. stearothermophilus), Bacillus atrophaeus (formerly B. subtilis), or Bacillus coagulans. The carriers should be individually packaged for use either within the package or for use upon removal from the package as an unpackaged biological indicator. The packaged biological indicator on the carrier has a particular labeled spore count of NLT 103 and NMT 109 spores/carrier. It has a survival time and kill time appropriate to the labeled spore count and to the decimal reduction value (D value, in min), specified by: Survival time (in min) = NLT (labeled D value) (log of labeled spore count per carrier 2), and Kill time (in min) = NMT (labeled D value) (log of labeled spore count per carrier + 4). IDENTIFICATION Identification for the biological indicator is of less importance than the more relevant concerns of population and
USP 32
resistance to the sterilization processes. The manufacturer should identify the species used. PERFORMANCE TESTS [NOTESee Biological IndicatorsResistance Performance Tests 55] D VALUE Analysis: Proceed as directed in the relevant procedure for D Value Determination. Acceptance criteria: The requirements of the test are met if the determined D value is within 20% of the labeled D value for the selected sterilizing conditions, and if the confidence limits of the estimate are within 10% of the determined D value. The D value determination method used should be that identified by the biological indicator manufacturer. SURVIVAL TIME and KILL TIME Analysis: Follow the procedure in the subsection Survival Time and Kill Time in D Value Determination. Acceptance criteria: The requirements of the test are met if all of the carriers subjected to sterilization exposure conditions intended to indicate survival show evidence of growth among the exposed carriers, while none of the carriers subjected to conditions designed to induce total kill show growth. If for either the survival test or the kill time test, NMT one carrier out of both groups fails the survival or kill requirements, continue the corresponding test with four additional groups, each consisting of 10 carriers, according to the procedure described. If all of the additional specimens subjected to the specific sterilization process either meet the survival requirements for the survival test time or meet the kill requirement for the kill test, whichever is applicable, the requirements are met. TOTAL VIABLE SPORE COUNT Analysis: Proceed as directed in the subsection Biological Indicators for Moist Heat, Dry Heat, and Gaseous Modes of Sterilization, Nonpaper Carriers in Total Viable Spore Count. Acceptance criteria: The requirements of the test are met if the average number of viable spores/carrier are within 50% and +300% of the labeled count/carrier or within a lesser range that may be stated by the manufacturer. 103109 spores/carrier of labeled spore count. SPECIFIC TESTS PURITY: There is no evidence of contamination with other microorganisms following examination of spores recovered from the carriers using a suitable plate culture medium. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in the original package under the conditions recommended on the label, and protect the package from light, toxic substances, excessive heat, and high relative humidity or moisture. The packaging or container materials do not adversely affect the performance of the article used as directed in the labeling. EXPIRATION DATE: The expiration date is determined on the basis of stability studies and is NMT 18 months from the date of manufacture. The date of manufacture is the date on which the first determination of the total viable count was made. DISPOSAL: Prior to destruction or discarding the carriers, sterilize by moist heat sterilization to ensure that the carrier surface is exposed to 121 for NLT 30 min, or by an equivalent method recommended by the manufacturer. LABELING: Label the package or package insert to state that it is a biological indicator prepared on a carrier for use in label-specified applications for moist heat, dry heat, and/or gaseous sterilization. State the biological indicator D value obtained under defined exposures to stated sterilization conditions using the Survival Curve method, SpearmanKarber method, or Stumbo-Murphy-Cochran method of D value analysis. State the survival time and kill time for the
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biological indicator carrier under specified conditions on the label. The total viable spore count per carrier following heat shock treatment must also appear on the label or package insert. State in the labeling the strain and ATCC number of the spore suspension used to inoculate the carriers and instructions for spore recovery and for safe disposal of the carriers. Indicate in the labeling that the stated D value is reproducible only under the exact conditions determined by the manufacturer and that the user would not necessarily obtain the same results if different exposure conditions were used. State that the user should determine the suitability of the carrier biological indicator for the users particular purpose and exposure conditions.
Biological Indicator for Steam Sterilization, Paper Carrier

(Comment on this Monograph)id=m9510=Biological Indicator for Steam Sterilization, Paper Carrier=B-Monos.pdf) DEFINITION Biological Indicator for Steam Sterilization, Paper Carrier, is a defined solution of viable spores made from a culture derived from a specified strain of Bacillus stearothermophilus, on a suitable grade of paper carrier, individually packaged in a suitable container readily penetrable by steam, and characterized for predictable resistance to steam sterilization. The packaged Biological Indicator for Steam Sterilization, Paper Carrier, has a particular labeled spore count per carrier of NLT 104 and NMT 109 spores. When labeled for and subjected to steam sterilization conditions at a particular temperature, it has a survival time and kill time appropriate to the labeled spore count and to the decimal reduction value (D Value, in min) of the solution, specified by: Survival time (in min) = NLT (labeled D Value) (log labeled spore count per carrier 2); and Kill time (in min) = NMT (labeled D Value) (log labeled spore count per carrier + 4) IDENTIFICATION The biological indicator organism complies substantially with the morphological, cultural, and biochemical characteristics of the strain of Bacillus stearothermophilus, ATCC No. 7953 or 12980, whichever is stated in the labeling: under microscopic examination it consists of Gram-positive rods with oval endospores in subterminally swollen cells; when incubated in nutrient broth for 17 h and used to inoculate appropriate solid media, growth occurs when the inoculated media are incubated aerobically for 24 h at 55 to 60, and similar inoculated media incubated concomitantly at 30 to 35 show no evidence of growth in the same period. When examined under conventional biochemical tests for microbial characterization, it shows a delayed weak positive catalase reaction, it does not utilize citrate, propionate or hippurate, it reduces nitrate, but it does not liquefy gelatin, and it gives a negative result with the Voges-Proskauer test. Organisms derived from ATCC strain No. 7953 show negative egg yolk and starch hydrolysis reactions, while those derived from ATCC strain No. 12980 show positive reactions in both tests. PERFORMANCE TESTS [NOTESee Biological IndicatorsResistance Performance Tests 55.] D VALUE Analysis: Proceed as directed for D Value, using the procedure for Biological Indicator for Steam Sterilization, Paper Carrier. Acceptance criteria: The determined D Value is within 20% of the labeled D Value for the selected sterilizing
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temperature, and the confidence limits of the estimate are within 10% of the determined D Value. SURVIVAL TIME and KILL TIME Analysis: Proceed as directed for Survival Time and Kill Time, using the procedure for Biological Indicator for Steam Sterilization, Paper Carrier. Acceptance criteria Survival time: All specimens show evidence of growth. Kill time: No specimen shows growth. If for either the Survival time test or the Kill time test, NMT 1 specimen out of both groups fails the survival requirement or the kill requirement (whichever is applicable), continue the corresponding test with four additional groups, each consisting of 20 specimens, according to the procedure described. If all of the additional specimens subjected to steam sterilization either meet the survival requirement for the Survival time test or meet the kill requirement for the Kill time test, whichever is applicable, the requirements of the test are met. TOTAL VIABLE SPORE COUNT Analysis: Proceed as directed for Total Viable Spore Count, using the procedure for Biological Indicator for Steam Sterilization, Paper Carrier. Acceptance criteria: The log average number of viable spores per carrier is NLT 0.3 log of the labeled spore count per carrier and does not exceed the log labeled spore count per carrier by 0.48. SPECIFIC TESTS PURITY Presence of contamination by other microorganisms: By examination of the spores on a suitable plate culture medium, there is no evidence of contamination with other microorganisms. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in the original package under the conditions recommended on the label, and protect it from light, toxic substances, excessive heat, and moisture. The packaging and container materials do not adversely affect the performance of the article used as directed in the labeling. EXPIRATION DATE: The expiration date is determined on the basis of stability studies and is NLT 18 months from the date of manufacture, the date of manufacture being the date on which the first determination of the total viable spore count was made. DISPOSAL: Prior to destruction or discard, sterilize it by steam at 121 for NLT 30 min, or by no less than an equivalent method recommended by the manufacturer. This includes a test strip employed in any test procedures for the strips themselves. LABELING: Label it to state that it is a Biological Indicator for Steam Sterilization, Paper Carrier; to indicate its D Value, the method used to determine such D Value, i.e., by spore count or fraction negative procedure after graded exposures to the sterilization conditions; the Survival time and Kill time under specified sterilization conditions stated on the label; its particular total viable spore count with a statement that such count has been determined after preliminary heat treatment; and its recommended storage conditions. State in the labeling the size of the paper carrier, the strain and ATCC number from which the spores were derived, and instructions for spore recovery and for safe disposal of the indicator. Indicate in the labeling that the stated D Value is reproducible only under the exact conditions under which it was determined, that the user would not necessarily obtain the same result and that the user should determine the suitability of the biological indicator for the particular use.
Biological Indicator for Steam Sterilization, Self-Contained

(Comment on this Monograph)id=m9512=Biological Indicator for Steam Sterilization, Self-Contained=B-Monos.pdf) DEFINITION Biological Indicator for Steam Sterilization, Self-Contained, is a Biological Indicator for Steam Sterilization, Paper Carrier individually packaged in a suitable container readily penetrable by steam and designed to hold an appropriate bacteriological culture medium, so as to enable the packaged carrier, after subjection to saturated steam sterilization conditions, to be incubated in the supplied medium in a self-contained system. The supplied medium may contain a suitable indicator as a convenience for determining by a color change whether or not spores have survived. The design of the self-contained system is such that, after exposure to the specified sterilization conditions and inoculation of the medium under closed conditions as stated in the labeling, there is no loss of medium and inoculum during subsequent transport and handling, if done according to the provided instructions. The materials of which the self-contained system are made are such that there is no retention or release of any substance that may cause inhibition of growth of surviving spores under the incubation conditions stated in the labeling. IDENTIFICATION The biological indicator organism complies substantially with the morphological, cultural, and biochemical characteristics of the strain of Bacillus stearothermophilus, ATCC No. 7953 or 12980, whichever is stated in the labeling: under microscopic examination it consists of Gram-positive rods with oval endospores in subterminally swollen cells; when incubated in nutrient broth for 17 h and used to inoculate appropriate solid media, growth occurs when the inoculated media are incubated aerobically for 24 h at 5560, and similar inoculated media incubated concomitantly at 3035 show no evidence of growth in the same period. When examined under conventional biochemical tests for microbial characterization, it shows a delayed weak positive catalase reaction; it does not utilize citrate, propionate, or hippurate; it reduces nitrate, but it does not liquefy gelatin; and it gives a negative result with the Voges-Proskauer test. Organisms derived from ATCC strain No. 7953 show negative egg yolk and starch hydrolysis reactions, while those derived from ATCC strain No. 12980 show positive reactions in both tests. PERFORMANCE TESTS [NOTESee Biological IndicatorsResistance Performance Tests 55.] D VALUE Analysis: Proceed as directed for the relevant procedure for D Value Determination. Acceptance criteria: The determined D value is within 20% of the labeled D value for the selected sterilizing temperature, and the confidence limits of the estimate are within 10% of the determined D value. SURVIVAL TIME and KILL TIME Analysis: Proceed as directed for Survival Time and Kill Time, using the procedure for Biological Indicator for Steam Sterilization, Self-Contained. Acceptance criteria Survival time: All specimens show evidence of growth. Kill time: No specimen shows growth. If for either the Survival time test or the Kill time test, NMT 1 specimen out of both groups fails the survival requirement or the kill requirement (whichever is applicable), continue the
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corresponding test with 4 additional groups, each consisting of 20 specimens, according to the procedure described. If all of the additional specimens subjected to steam sterilization either meet the survival requirement for the Survival time test or meet the kill requirement for the Kill time test, whichever is applicable, the requirements of the test are met. TOTAL VIABLE SPORE COUNT Analysis: Proceed as directed for Total Viable Spore Count, using the procedure for Biological Indicator for Steam Sterilization, Paper Carrier. Acceptance criteria: The average number of viable spores per carrier is NLT 0.3 log of the labeled spore count per carrier and does not exceed the log labeled spore count per carrier by 0.48. SPECIFIC TESTS MEDIUM SUITABILITY Sterility: Incubate 10 self-contained biological indicator systems at 5560, or at the optimal recovery temperature specified by the manufacturer, for 48 h, making sure that there is no contact between the individual spore strips and the supplied medium. Examine the incubated medium visually (for change in color indicator or turbidity) and microscopically (for absence of microbial growth). Growth promotion of medium prior to sterilization treatment Analysis: Submerge 10 self-contained units in a water bath maintained at 95100 for 15 min. Start timing when the temperature of the container contents reach 95. Cool rapidly in an ice-water bath (04). Remove the units from the ice-water bath, submerge each spore strip with the selfcontained medium, incubate at 5560, or at the optimal recovery temperature specified by the manufacturer, and examine visually after 48 h for growth (for turbidity or change in color), and microscopically (for microbial growth). Acceptance criteria: All the specimens under test show growth. If one or more of the specimens do not show growth, repeat the test with 20 additional units. The additional units all show growth. Growth promotion of medium after exposure to sterilization conditions: Expose the specified number of units for both the Survival time and Kill time stated in the labeling, as described in the section Biological Indicator for Steam Sterilization, Self-Contained under Biological IndicatorsResistance Performance Tests 55. Incubate the spore strips submerged in the self-contained medium according to the instructions of the manufacturer. At the end of the incubation period confirm the existence of growth in each of the specimens that were exposed for each Survival time and the absence of growth in each of the specimens that were exposed for each Kill time by visual inspection (turbidity or color indicator change) and by separate microscopic examination of each specimen and confirm, where applicable, correspondence of the labeled color to the appearance of growth in the supplied medium. Ability of medium to support growth after exposure to the sterilization conditions Analysis: Take a stated number of units (e.g., 10) after they have been exposed for each Kill time stated in the labeling as directed in the preceding section. Aseptically remove and pool the medium from each unit. Prepare a suspension of the indicator microorganism as directed for Total Viable Spore Counts under Biological Indicator for Steam Sterilization, Paper Carrier. Prepare a dilution of that suspension so as to contain 1001000 viable
Official Monographs / Biotin 85

microorganisms in 1 mL. Inoculate the pooled medium with enough suspension to contain a total of 1001000 microorganisms in a 10 mL aliquot of NMT the volume from 10 units of the pooled medium. Incubate the inoculated pooled medium as directed for Total Viable Spore Count. Acceptance criteria: Clear evidence of growth is obtained within 7 days. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in the original package under the conditions recommended on the label, and protect from light, from substances that may adversely affect the contained microorganisms, from excessive heat, and from moisture. EXPIRATION DATE: The expiration date is determined on the basis of stability studies and is NLT 18 months from the date of manufacture, the date of manufacture being the date on which the first determination of the total viable spore count was made. DISPOSAL: Prior to destruction or discard, sterilize it by steam at 121 for NLT 30 min, or by NLT an equivalent method recommended by the manufacturer. This includes test strips employed in any test procedures for the strips themselves. LABELING: Label it to state that it is a Biological Indicator for Steam Sterilization, Self-Contained; to indicate the D value of the self-contained system, the method used to determine such D value (i.e., by spore count or fraction negative procedure after graded exposures to the sterilization conditions); the Survival time and Kill time under the specified conditions stated on the label; its particular total viable spore count, with a statement that such count has been determined after preliminary heat treatment; and its recommended storage conditions. State on the labeling that the supplied bacteriological medium will meet requirements for growth-promoting ability, the strain and ATCC number from which the spores were derived, and the instructions for spore recovery and for safe disposal of the indicator unit. Also indicate in the labeling that the stated resistance characteristics are reproducible only under steam sterilization conditions at the stated temperature and only under the exact conditions under which it was determined, that the user would not necessarily obtain the same result, and that the user should determine the suitability of the biological indicator for the particular use.
Biotin
(Comment on this Monograph)id=m9515=Biotin=B-Monos.pdf)
244.31 C10H16N2O3S 1H-Thieno3,4-dimidazole-4-pentanoic acid, hexahydro-2-oxo-, 3aS-(3a,4,6a)-; (3aS,4S,6aR)-Hexahydro-2-oxo-1H-thieno3,4-dimidazole-4-valeric acid [58-85-5]. DEFINITION Biotin contains NLT 97.5% and NMT 100.5% of C10H16N2O3S.
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ASSAY PROCEDURE Sample: 500 mg Analysis: Dissolve the Sample in 20 mL of benzene, add 2 drops of crystal violet TS, and titrate with 0.1 N perchloric acid VS to a blue endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 31.15 mg of C21H29NO. Acceptance criteria: NLT 98.0% and NMT 101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Standard solution: Prepare in methanol as directed. Sample solution: Prepare in methanol as directed. Eluant: A mixture of methanol and ammonium hydroxide (100:1.5) Visualization: 17 Analysis: Proceed as directed. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: Between 112 and 116 LOSS ON DRYING 731 Analysis: Dry a sample at 105 for 3 h. Acceptance criteria: The sample loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Biperiden RS
IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 500 mg of Biotin Analysis: Mix the sample with 100 mL of water. Add phenolphthalein TS and titrate the suspension slowly with 0.1 N sodium hydroxide VS, while heating and stirring continuously. Each mL of 0.1 N sodium hydroxide is equivalent to 24.43 mg of C10H16N2O3S. Acceptance criteria: NLT 97.5% and NMT 100.5% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S Sample solution: 20 mg/mL in 0.1 N sodium hydroxide Acceptance criteria: Between +89 and +93 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Store in tight containers. USP REFERENCE STANDARDS 11 USP Biotin RS
Biperiden
(Comment on this Monograph)id=m9530=Biperiden=BMonos.pdf)
311.46 C21H29NO 1-Piperidinepropanol, -bicyclo[2.2.1]hept-5-en-2-yl--phenyl-; -5-Norbornen-2-yl--phenyl-1-piperidinepropanol [514-65-8]. DEFINITION Biperiden contains NLT 98.0% and NMT 101.0% of C21H29NO, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Solution: Transfer 180 mg of biperiden to a 200-mL volumetric flask, add 1 mL of lactic acid and dilute with water to volume (0.9 mg/mL). Acceptance criteria: Absorptivities at 257 nm, calculated on the dried basis, do not differ by more than 3.0%. C. PROCEDURE Sample: 20 mg Analysis: Dissolve the Sample in 5 mL of phosphoric acid. Acceptance criteria: A green color is produced. D. PROCEDURE Sample solution: Dissolve 200 mg in 80 mL of water with the aid of 0.5 mL of 3 N hydrochloric acid, warming, if necessary, to effect solution, and then cool. Analysis: To 5 mL of Sample solution, add 1 drop of hydrochloric acid and several drops of mercuric chloride TS. Acceptance criteria: A white precipitate is formed. E. PROCEDURE Sample solution: Use Sample solution from Procedure D Analysis: To a second 5-mL portion of the Sample solution, add bromine TS dropwise. Acceptance criteria: A yellow precipitate forms, which redissolves on shaking, and finally, upon the addition of more bromine TS, a permanent precipitate is formed.
Biperiden Hydrochloride
(Comment on this Monograph)id=m9550=Biperiden Hydrochloride=B-Monos.pdf) C21H29NO HCl 347.92 1-Piperidinepropanol, -bicyclo[2.2.1]hept-5-en-2-yl--phenyl-, hydrochloride; -5-Norbornen-2-yl--phenyl-1-piperidinepropanol hydrochloride [1235-82-1]. DEFINITION Biperiden Hydrochloride contains NLT 98.0% and NMT 101.0% of C21H29NO HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Solution: 1 mg/mL Medium: Methanol Acceptance criteria: Absorptivities at 257 nm do not differ by more than 3.0%. C. PROCEDURE Sample: 20 mg Analysis: Dissolve the Sample in 5 mL of phosphoric acid. Acceptance criteria: A green color is produced. D. PROCEDURE Sample solution: 1 in 500 Analysis: To a 5-mL portion of the Sample solution add bromine TS dropwise.
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Acceptance criteria: A yellow precipitate, which dissolves on shaking, is formed. Addition of more bromine TS produces a precipitate that does not dissolve on shaking. E. IDENTIFICATION TESTSGENERAL, Chloride 191 Sample solution: 1 in 500 Acceptance criteria: Meets the requirements of the tests ASSAY PROCEDURE Sample: 500 mg Analysis: Dissolve the Sample in 80 mL of glacial acetic acid, warming slightly, if necessary, to effect solution. Cool, add 1 drop of crystal violet TS and 10 mL of mercuric acetate TS, and titrate with 0.1 N perchloric acid VS to a blue endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 34.79 mg of C21H29NO HCl. Acceptance criteria: NLT 98.0% and NMT 101.0% IMPURITIES Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Standard solution: Prepare in methanol as directed. Sample solution: Prepare in methanol as directed. Eluant: A mixture of methanol and ammonium hydroxide (100:1.5) Visualization: 17 Analysis: Proceed as directed. SPECIFIC TESTS LOSS ON DRYING 731 Analysis: Dry a sample at 105 for 3 h. Acceptance criteria: The sample loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Biperiden Hydrochloride RS
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Visualization: Iodine vapor, 10 min Analysis: Separately apply the Sample solution and the Standard solution to the chromatographic plate. Allow the applications to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by exposing the plate for 10 min to iodine vapors in a pre-equilibrated closed chamber, on the bottom of which there are iodine crystals. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Solution A: 38 mg/mL of monobasic sodium phosphate and 2 mg/mL of anhydrous dibasic sodium phosphate in water. Adjust to a pH of 5.3 0.1, if necessary. Solution B: Dissolve 400 mg of bromocresol purple in 30 mL of water, add 6.3 mL of 0.1 N sodium hydroxide, and dilute with water to 500 mL. Phosphate bufferbromocresol purple solution: Mix equal volumes of Solution A, Solution B, and chloroform, shake in a separator, and discard the chloroform. If appreciable color is extracted, repeat with additional portions of chloroform until no color is extracted. Standard stock solution: 0.8 mg/mL of USP Biperiden Hydrochloride RS in methanol Standard solution: 40 g/mL of USP Biperiden Hydrochloride RS from Standard stock solution: transfer 5.0 mL of Standard stock solution to a 100-mL volumetric flask, add 25 mL of water, and dilute with methanol to volume. Sample solution: Transfer a portion of finely powdered Tablets, equivalent to 2 mg of biperiden hydrochloride, from NLT 20 Tablets, to a 50-mL volumetric flask, add 12.5 mL of water, and heat on a steam bath for 15 min. Cool, and dilute with methanol to volume. Blank: Methanol and water (3:1) Analysis Samples: Standard solution, Sample solution, and Blank Transfer 5 mL each of the Standard solution, the Sample solution, and the Blank to individual separators, each containing 10 mL of Phosphate bufferbromocresol purple solution. Extract the solution in each separator with 20 mL of chloroform for 2 min. After the layers have separated, pass each chloroform extract through filter paper (Whatman No. 31 or equivalent) into separate glassstoppered, 50-mL volumetric flasks. In the same manner, extract the solution in each separator with another 20-mL portion of chloroform, filter, and wash each filter with 8 mL of chloroform, collecting each combined filtrate and washing, respectively, in the 50-mL volumetric flask containing the first extract. Dilute each with chloroform to volume. Concomitantly determine the absorbances of the solutions in 1-cm cells at the wavelength of maximum absorbance at about 408 nm, with a suitable spectrophotometer, using the Blank to set the instrument. Calculate the percentage of C21H29NO HCl in the portion of Tablets taken: Result = (AU/AS) (CS/CU) F 100/L AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Biperiden Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of the Sample solution (g/mL)
Biperiden Hydrochloride Tablets

(Comment on this Monograph)id=m9580=Biperiden Hydrochloride Tablets=B-Monos.pdf) DEFINITION Biperiden Hydrochloride Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of C21H29NO HCl. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture. Condition by heating the plate at 105 for 1 h and allowing to cool. Standard solution: Proceed as directed for the Sample solution using 10 mg of USP Biperiden Hydrochloride RS in place of the powdered Tablets. Sample solution: To a quantity of finely powdered Tablets, equivalent to 10 mg of biperiden hydrochloride, add 5 mL of water, mix, and sonicate to disperse the powder. Add 5 mL of methanol to the flask, mix, and sonicate for 15 min. Filter the solution into a separator, add 2 mL of 1 N sodium hydroxide and 10 mL of chloroform, and shake for 3 min. Filter the chloroform layer into a stoppered flask, and use the chloroform filtrate as the Sample solution. Application volume: 20 L Developing solvent system: Methanol and ammonium hydroxide (100:1.5)
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IDENTIFICATION IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Sample solution: A volume of Injection equivalent to 50 mg of biperiden lactate Standard solution: 50 mg of USP Biperiden RS in 25 mL of 0.01 N hydrochloric acid Analysis: Proceed as directed under IdentificationOrganic Nitrogenous Bases 181, beginning with Transfer the liquid to a separator. Acceptance criteria: Meets the requirements ASSAY PROCEDURE Solution A: 38 mg/mL of monobasic sodium phosphate and 2 mg/mL of anhydrous dibasic sodium phosphate in water Adjust the pH of the solution to 5.3 0.1, if necessary. Solution B: Dissolve 400 mg of bromocresol purple in 30 mL of water. Add 6.3 mL of 0.1 N sodium hydroxide, and dilute with water to 500 mL. Phosphate bufferbromocresol purple solution: Mix equal volumes of Solution A, Solution B, and chloroform. Shake in a separator, and discard the chloroform. If appreciable color is extracted, repeat with additional portions of chloroform until no color is extracted. Standard stock solution: 0.8 mg/mL of USP Biperiden RS in methanol Standard solution: 40 g/mL of USP Biperiden RS from Standard stock solution Transfer 5.0 mL of Standard stock solution to a 100-mL volumetric flask, add 25 mL of water, and dilute with methanol to volume. Sample solution: Transfer an amount of the Injection, equivalent to 5 mg of biperiden lactate, to a 100-mL volumetric flask. Add 25 mL of water, and dilute to volume with methanol. Blank: Methanol and water (3:1) Analysis Samples: Standard solution, Sample solution, and Blank Transfer 5.0 mL each of the Standard solution, the Sample solution, and the Blank to individual separators, each containing 10.0 mL of Phosphate bufferbromocresol purple solution. Extract the solution in each separator with 20.0 mL of chloroform for 2 min. After the layers have separated, pass each chloroform extract through filter paper (Whatman No. 31 or equivalent) into separate glass-stoppered, 50-mL volumetric flasks. In the same manner, extract the solution in each separator with another 20.0-mL portion of chloroform, filter, and wash each filter with 8 mL of chloroform, collecting each combined filtrate and washing, respectively, in the 50-mL volumetric flask containing the first extract. Dilute each with chloroform to volume. Concomitantly determine the absorbances of the solutions in 1-cm cells at the wavelength of maximum absorbance at about 408 nm, with a suitable spectrophotometer, using the blank to set the instrument. Calculate the percentage of C21H29NO C3H6O3 in the portion of Injection taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) F 100 AU AS CS CU Mr1 = absorbance from the Sample solution = absorbance from the Standard solution = concentration of USP Biperiden Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of biperiden lactate in the Sample solution (mg/mL) = molecular weight of biperiden lactate, 401.55
F = conversion factor, 0.001 (g to mg) Acceptance criteria: NLT 93.0% and NMT 107.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 45 min [NOTEDetermine the amount of C21H29NO HCl dissolved by employing the following method.] Phosphate bufferbromocresol purple solution: Prepare as directed for Assay. Standard stock solution: 0.8 mg/mL of USP Biperiden Hydrochloride RS in methanol Standard solution: 2 g/mL of USP Biperiden Hydrochloride RS, prepared as follows: Pipet 5 mL of Standard stock solution into a 500-mL volumetric flask, and add 0.01 N hydrochloric acid to volume. Pipet 25 mL of this solution into a suitable beaker, and adjust with 0.01 N sodium hydroxide to a pH of 5.3. Transfer this solution to a 100-mL volumetric flask with the aid of water, and dilute with water to volume. Sample solution: Sample per Dissolution 711. Filter 75 mL of the solution under test, pipet 50 mL of the clear filtrate into a suitable beaker, and adjust with 0.01 N sodium hydroxide to a pH of 5.3. Transfer this solution to a 100-mL volumetric flask with the aid of water, and dilute with water to volume. Blank: Water Analysis Samples: Standard solution, Sample solution, and Blank Pipet 20 mL each of the Standard solution, the Sample solution, and the Blank into individual separators, each containing 10.0 mL of Phosphate bufferbromocresol purple solution. Extract the solution in each separator with 40.0 mL of chloroform for 10 min. After the layers have separated, pass each chloroform extract through filter paper into separate, glass-stoppered containers, discarding the first 10 mL of each filtrate. Determine the amount of C21H29NO HCl dissolved from absorbances at the wavelength of maximum absorbance at about 408 nm (10-cm cells) of the extract from the Sample solution in comparison with that of the extract from the Standard solution, using the Blank to set the instrument. Acceptance criteria: NLT 75% (Q) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Biperiden Hydrochloride RS
Biperiden Lactate Injection

(Comment on this Monograph)id=m9610=Biperiden Lactate Injection=B-Monos.pdf) 401.54 C21H29NO C3H6O3 1-Piperidinepropanol, -bicyclo[2.2.1]hept-5-en-2-yl--phenyl-, compd. with 2-hydroxypropanoic acid (1:1); -5-Norbornen-2-yl--phenyl-1-piperidinepropanol lactate (salt) [7085-45-2]. DEFINITION Biperiden Lactate Injection is a sterile solution of biperiden lactate (C21H29NO C3H6O3) in Water for Injection, prepared from Biperiden with the aid of Lactic Acid. It contains NLT 95.0% and NMT 105.0% of the labeled amount of C21H29NO C3H6O3.
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Mr2 = molecular weight of biperiden, 311.47 F = conversion factor, 0.001 (g to mg) Acceptance criteria: 95.0%105.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 83.3 USP Endotoxin Units/mg of biperiden lactate PH 791: 4.85.8 OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Biperiden RS USP Endotoxin RS
Official Monographs / Bisacodyl 89

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Bisacodyl RS
Bisacodyl Suppositories
(Comment on this Monograph)id=m9685=Bisacodyl Suppositories=B-Monos.pdf) DEFINITION Bisacodyl Suppositories contain NLT 90.0% and NMT 110.0% of the labeled amount of C22H19NO4. IDENTIFICATION A. PROCEDURE Sample: Transfer the equivalent to 150 mg of bisacodyl from Suppositories, to a 500-mL conical flask, add 75 mL of solvent hexane, and heat on a steam bath until melted. Filter the solution, with the aid of a vacuum, through a medium-porosity, sintered-glass funnel, and wash the residue with about 100 mL of warm solvent hexane until it is free from fat. Continue the vacuum until the residue appears dry. Dissolve the residue by rinsing the filter with about 50 mL of warm acetone, collecting the filtrate in a 150-mL beaker, and evaporating the filtrate on a steam bath to a volume of about 5 mL. To the residual liquid add about 75 mL of water, heat on a steam bath for 15 min, and cool. Scratch the sides of the beaker to induce crystallization, filter the crystals, and dry at 100 for about 15 min. Analysis 1: The bisacodyl so obtained melts between 129 and 135. Analysis 2: Infrared Absorption 197S Cell: 1.0 mm Sample solution: Dissolve a portion of the crystalline Sample in chloroform (1 in 200). B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.074 M sodium acetate in water, adjusted with 2.5% acetic acid to a pH of 7.4 Mobile phase: Acetonitrile and Solution A (9:11) Standard solution: 0.5 mg/mL of USP Bisacodyl RS in acetonitrile Sample solution: To the equivalent to 100 mg of bisacodyl from Suppositories, in a 500-mL separator, add 150 mL of n-hexane, and shake until all the suppositories are dissolved. Add 50 mL of acetonitrile, shake for 1 min, and allow the layers to separate. Drain the lower layer into a 200-mL volumetric flask, and extract the n-hexane layer remaining in the separator with two 50-mL portions of acetonitrile, combining the lower layers in the volumetric flask. Dilute the combined extracts in the volumetric flask with acetonitrile to volume, mix, and filter. Chromatographic system (See Chromatography 621, System Suitability.)
Bisacodyl
(Comment on this Monograph)id=m9660=Bisacodyl=BMonos.pdf)
361.39 C22H19NO4 Phenol, 4,4-(2-pyridinylmethylene)bis-, diacetate (ester); 4,4-(2-Pyridylmethylene)diphenol diacetate (ester). [603-50-9]. DEFINITION Bisacodyl contains NLT 98.0% and NMT 101.0% of C22H19NO4, calculated on the dried basis. [CAUTIONAvoid inhalation and contact with the eyes, skin, and mucous membranes.] IDENTIFICATION A. INFRARED ABSORPTION 197S Cell: 1.0 mm Solution: 1 in 200 solution in chloroform, previously dried B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 263 nm Solution: 20 g/mL in 0.05 N hydrochloric acid Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0%. ASSAY PROCEDURE Sample solution: Dissolve 250 mg of Bisacodyl in 70 mL of glacial acetic acid, and add 3 drops of p-naphtholbenzein TS. Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 36.14 mg of C22H19NO4. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 131135 LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.5% of its weight.
90
Bisacodyl / Official Monographs
USP 32
Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for bisacodyl and ethylparaben are about 2.0 and 1.0, respectively.] Suitability requirements Resolution: NLT 7.0 between bisacodyl and the internal standard Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 1.2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percent label claim of C22H19NO4: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the bisacodyl peak to the internal standard peak from the Sample solution = peak response ratio of the bisacodyl peak to the RS internal standard peak from the Standard solution = concentration of USP Bisacodyl RS in the CS Standard solution (g/mL) CU = concentration of bisacodyl in the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% RU SPECIFIC TESTS PH 791: 5.06.8 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in unit-dose containers at a temperature not exceeding 30. USP REFERENCE STANDARDS 11 USP Bisacodyl RS
Mode: LC Detector: UV 265 nm Guard column: Packing L2 Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% of replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H19NO4 taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bisacodyl RS in the Standard solution (mg/mL) = concentration of bisacodyl in the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers at a temperature not exceeding 30. USP REFERENCE STANDARDS 11 USP Bisacodyl RS
Bisacodyl Rectal Suspension

(Comment on this Monograph)id=m9680=Bisacodyl Rectal Suspension=B-Monos.pdf) DEFINITION Bisacodyl Rectal Suspension is a suspension of Bisacodyl in a suitable aqueous medium. It contains NLT 90.0% and NMT 115.0% of the labeled amount of C22H19NO4. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol and 0.01 M monobasic potassium phosphate (3:2) Internal standard solution: Dissolve ethylparaben in methanol and dilute with an equal volume of water (5.0 mg/mL). Standard solution: Dissolve a quantity of USP Bisacodyl RS in methanol, add a volume of Internal standard solution, and dilute with methanol to obtain a solution of 67 g/mL of bisacodyl and 250 g/mL of ethylparaben. Sample solution: Transfer a measured volume of Rectal Suspension equivalent to 6.7 mg of bisacodyl to a 100-mL volumetric flask. Add 5.0 mL Internal standard solution and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.)
Bisacodyl Delayed-Release Tablets

(Comment on this Monograph)id=m9695=Bisacodyl DelayedRelease Tablets=B-Monos.pdf) DEFINITION Bisacodyl Delayed-Release Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C22H19NO4. Bisacodyl Delayed-Release Tablets are enteric coated. IDENTIFICATION A. INFRARED ABSORPTION 197S Cell: 1.0 mm Sample solution: Macerate the equivalent of 300 mg of bisacodyl from powdered Tablets, with 100 mL of acetone. Heat on a steam bath to boiling, filter, and evaporate to about 20 mL. Add 200 mL of water, and warm the mixture on the steam bath, passing a stream of nitrogen over the surface to evaporate the acetone. After 30 min, cool the mixture, and filter through a sintered-glass funnel. Discard the filtrate, and dissolve the crystals in 50 mL of acetone. Evaporate the solution to about 15 mL, add about 75 mL of water, heat on a steam bath for 15 min, and then cool.
USP 32
Scratch the sides of the beaker to induce crystallization, filter the crystals, and dry at 100 for about 15 min. Using the crystals so obtained prepare a solution (1 in 200) in chloroform. B. The retention time of the major peak for bisacodyl of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.074 M sodium acetate in water, adjusted with 2.5% acetic acid to a pH of 7.4 Mobile phase: Acetonitrile and Solution A (9:11) Standard solution: 0.5 mg/mL of USP Bisacodyl RS in acetonitrile Sample solution: Equivalent to 100 mg of bisacodyl from powdered Tablets, in a 200-mL volumetric flask, add 25 mL of water, and shake by mechanical means for 15 min followed by sonication for 15 min. Add 100 mL of acetonitrile, and shake by mechanical means for 15 min followed by sonication for 15 min. Dilute with acetonitrile to volume, mix, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Guard column: Packing L2 Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percent label claim of C22H19NO4: Result = (rU/rs) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bisacodyl RS in the Standard solution (mg/mL) CU = concentration of bisacodyl in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISINTEGRATION 701: Proceed as directed for DelayedRelease (Enteric Coated) Tablets: the tablets do not disintegrate after 1 h of agitation in simulated gastric fluid TS, but then disintegrate within 45 min in simulated intestinal fluid TS. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE Preserve in well-closed containers at a temperature not exceeding 30. LABELING: Label the Tablets to indicate that they are enteric coated. USP REFERENCE STANDARDS 11 USP Bisacodyl RS
Official Monographs / Bismuth 91
Milk of Bismuth
(Comment on this Monograph)id=m9750=Milk of Bismuth=BMonos.pdf) DEFINITION Milk of Bismuth contains bismuth hydroxide and bismuth subcarbonate in suspension in water, and yields NLT 5.2% and NMT 5.8% (w/w) of bismuth trioxide (Bi2O3).
Bismuth Subnitrate Nitric Acid Ammonium Carbonate Strong Ammonia Solution Purified Water, each To make 80 g 120 mL 10 g A sufficient quantity 1000 mL
Mix the Bismuth Subnitrate with 60 mL of Purified Water and 60 mL of the Nitric Acid in a suitable container, and agitate, warming gently until solution is effected. Pour this solution, with constant stirring, into 5000 mL of Purified Water containing 60 mL of the Nitric Acid. Dilute 160 mL of Strong Ammonia Solution with 4300 mL of Purified Water in a glazed or glass vessel of at least 12-L capacity. Dissolve the Ammonium Carbonate in this solution, and then pour the bismuth solution quickly into it with constant stirring. Add sufficient 6 N ammonium hydroxide, if necessary, to render the mixture distinctly alkaline, allow to stand until the precipitate has settled, then pour or siphon off the supernatant, and wash the precipitate twice with Purified Water, by decantation. Transfer the magma to a strainer of close texture, so as to provide continuous washing with Purified Water, the outlet tube being elevated to prevent the surface of the magma from becoming dry. When the washings no longer yield a pink color with phenolphthalein TS, drain the moist solution, transfer to a graduated vessel, add sufficient Purified Water to make 1000 mL, and mix. [NOTEThis method of solution may be varied, provided the product meets the following requirements.] IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Bismuth 191 and Carbonate 191: It meets the requirements. B. Add 1 mL of 3 N hydrochloric acid to 1 mL of Milk of Bismuth: a clear solution is produced. Pour the clear solution into 10 volumes of water: a white precipitate is formed. ASSAY PROCEDURE Sample solution: Evaporate a quantity of Milk of Bismuth to dryness, and ignite the residue to constant weight. From the weight of the Bi2O3 so obtained determine the percentage in the Assay sample. Acceptance criteria: 5.2%5.8% IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Sample solution: Evaporate 3.75 mL on a steam bath to dryness, add 2 mL of sulfuric acid, and heat until copious fumes of sulfur trioxide are evolved.
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Bismuth / Official Monographs
USP 32
Allow to cool, add 2 mL of nitric acid to the residue, dropwise, and warm until complete solution has been effected. Add about 60 mL of water and 0.3 mL of xylenol orange TS. Analysis Sample: Sample solution Titrate with 0.05 N edetate disodium VS to a yellow endpoint. Each mL of 0.05 N edetate disodium is equivalent to 10.45 mg of Bi. Acceptance criteria: 49%54% IMPURITIES Inorganic Impurities ARSENIC, Method I 211 Sample: 300 mg of Bismuth Citrate Analysis: Triturate Sample with an equal weight of calcium hydroxide, and ignite. Dissolve the residue in 5 mL of 3 N hydrochloric acid. Acceptance criteria: NMT 10 ppm LIMIT OF NITRATE Sample solution: The second portion of the cooled solution reserved from Identification test C Analysis: To the Sample solution, add an equal volume of sulfuric acid, and allow to cool. Into the liquid, drop a crystal of ferrous sulfate, and allow to stand for 30 min. Acceptance criteria: No brown or brownish black color appears around the crystal. LIMIT OF COPPER, LEAD, AND SILVER Standard solution: Prepare a solution containing 1000 g/mL of copper, a solution containing 1000 g/mL of lead, and a solution containing 1000 g/mL of silver. Transfer 3.0 mL of each solution to a 2000-mL volumetric flask, dilute with 1 N nitric acid to volume. [NOTEThe concentrations of copper, lead, and silver in this solution may be modified by using a different quantity or by further dilution to bring the absorption responses within the working range of the atomic absorption spectrophotometer.] Sample solution: Ignite 3 g of Bismuth Citrate, in a porcelain crucible, cool, and cautiously add 6 N nitric acid to dissolve the residue. Add 100 mL of water. A white precipitate forms. Filter this mixture, evaporate on a steam bath to obtain about 15 mL of solution, and filter again. Dilute the filtrate with water to 20.0 mL. Analysis Samples: Standard solution and Sample solution Concomitantly determine the absorbances of the Standard solution and the Sample solution at the emission lines of 324.7, 217, and 328.1 nm for copper, lead, and silver, respectively, with an atomic absorption spectrophotometer (see Spectrophotometry and LightScattering 851) equipped with copper, lead, and silver hollow-cathode lamps and an oxidizing flame. Acceptance criteria: The absorbances of the Sample solution do not exceed those of the Standard solution for each element (NMT 10 ppm). LIMIT OF SOLUBLE BISMUTH Standard solution: 242.0 mg of bismuth nitrate pentahydrate to a 100-mL volumetric flask. Add 3 mL of 1.5 N nitric acid, swirl to dissolve, and dilute with water to volume. Transfer 1.0 mL of this solution to a 500-mL volumetric flask, add 250 mL of 1.5 N nitric acid, and dilute with water to volume. This solution contains 2.0 g/mL of Bi. [NOTEThe concentration of bismuth in this solution may be modified by using a different quantity or by further dilution to bring the absorption responses within the working range of the atomic absorption spectrophotometer.] Sample solution: Prepare a mixture of 5.0 g of Bismuth Citrate in 100 mL of water, and stir by mechanical means
Acceptance criteria: NMT 0.8 ppm LEAD: To 5 mL add warm nitric acid, dropwise, until it is just dissolved, and pour the solution into 50 mL of water: a white precipitate may form. Filter, if necessary, evaporate the filtrate on a steam bath to 15 mL, again filter, and to 10 mL of the filtrate add an equal volume of 2 N sulfuric acid. Acceptance criteria: No precipitate is formed. LIMIT OF ALKALIES AND ALKALINE EARTHS Sample solution: 2.0 mL in 5 mL of hydrochloric acid, dilute with water to 100 mL, add hydrogen sulfide to precipitate the bismuth completely, and filter Analysis: To 50 mL of the clear filtrate add 5 drops of sulfuric acid, evaporate to dryness, and ignite. Acceptance criteria: The weight of the residue does not exceed 3 mg (0.3%). WATER-SOLUBLE SUBSTANCES: Boil 10 mL with 90 mL of water for 10 min, cool, add water to make the total volume 100 mL, mix, and filter. Evaporate 50 mL of the filtrate to dryness, and ignite it gently: the weight of the residue does not exceed 5 mg (0.1%). SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: The total bacterial count does not exceed 100 cfu/mL and the test for Escherichia coli is negative. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and protect from freezing.
Bismuth Citrate
(Comment on this Monograph)id=m9755=Bismuth Citrate=BMonos.pdf)
BiC6H5O7 [813-93-4].
398.08
DEFINITION Bismuth Citrate contains NLT 49% and NMT 54% of bismuth (Bi). IDENTIFICATION A. INFRARED ABSORPTION 197K: On the undried specimen B. When strongly heated, the salt chars, and on ignition leaves a more or less blackened residue having a yellow surface. The residue is soluble in warm nitric acid, and this solution, when dropped into a large excess of water, produces a white turbidity. C. PROCEDURE Sample solution: 1 g in ammonia TS Analysis: When treated with hydrogen sulfide in excess, a black precipitate is obtained. Filter this mixture, drive off the excess hydrogen sulfide by heating, and allow to cool. To a portion of this cooled solution add an excess of calcium hydroxide TS, and boil. Acceptance criteria: A white precipitate is formed. Reserve a second portion of the cooled solution for the test for Limit of nitrate. ASSAY PROCEDURE Sample solution: Transfer 300 mg of Bismuth Citrate to a porcelain crucible, and ignite.
USP 32
the suspension thus obtained for 2 h. Pass through filter paper. Pass the filtrate thus obtained through a filter having a 0.1-m or finer porosity. To 10.0 mL of the filtrate add 0.1 mL of nitric acid. Analysis Samples: Standard solution and Sample solution Concomitantly determine the absorbances of the Standard solution and the Sample solution at the emission line of 223.06 nm for bismuth with an atomic absorption spectrophotometer (see Spectrophotometry and LightScattering 851) equipped with a bismuth hollowcathode lamp and an oxidizing flame. Acceptance criteria: The absorbances of the Sample solution do not exceed those of the Standard solution (NMT 40 ppm). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, store at controlled room temperature, and prevent exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Bismuth Citrate RS

Analysis: To the Standard solution and the Sample solution, add 0.05 mL of Indigo carmine titrant. Carefully add 30 mL of sulfuric acid, and immediately titrate with Indigo carmine titrant to a stable blue endpoint. Acceptance criteria: The volume of Indigo carmine titrant consumed by the Sample solution does not exceed that consumed by the Standard solution (0.4%). LIMIT OF SILVER Standard solution: 7.87 g/mL of silver nitrate Sample solution: 2.0 g of Bismuth Subcarbonate, add 1 mL of water and 4 mL of nitric acid Analysis: Heat gently to achieve dissolution, add water to obtain 11 mL of solution, and cool. Add 2 mL of 1 N hydrochloric acid, and allow to stand in a dark place for 5 min. Acceptance criteria: No more turbidity is produced than corresponds to that produced with 10 mL of the Standard solution concomitantly treated with 1 mL of nitric acid and 2 mL of 1 N hydrochloric acid (0.0025%). LIMIT OF COPPER Standard stock solution: 1.34 g of cupric chloride, 10 g of ammonium chloride, and 3 mL of sodium metabisulfite solution (275 mg/mL) to 100.0 mL. This stock solution contains the equivalent of 5 mg/mL of copper. Standard solution: equivalent of 10 g/mL of copper made from the Standard stock solution in 2 N nitric acid. Mix 0.25 mL of this solution and 9.75 mL of water. Sample solution: 5 mL of the Sample stock solution retained from the test for Chloride, add 2 mL of 6 N ammonium hydroxide, dilute with water to 50 mL, mix, and filter. Analysis: To 10 mL each of the Standard solution and the Sample solution, add 1 mL of a solution of sodium diethyldithiocarbamate (1 in 1000). Acceptance criteria: No more color is obtained from the Sample solution than is obtained from the Standard solution (0.005%). LIMIT OF LEAD Diluent: 6 N nitric acid, lead-free Standard stock solution: 0.1598 mg/mL of lead nitrate in Diluent [NOTEThis solution contains 100 g/mL of lead.] Standard solution: 1.0 g/mL, 2.0 g/mL, and 3.0 g/mL of lead from Standard stock solution in Diluent Sample solution: 12.5 g of Bismuth Subcarbonate in 75 mL of Diluent. Heat to boiling for 1 min, cool, and dilute with water to 100 mL. Analysis: Concomitantly determine the absorbances of the Standard solutions and the Sample solution at the lead emission line of 283.3 nm with an atomic absorption spectrophotometer (See Spectrophotometry and Lightscattering 851) equipped with a lead hollow-cathode lamp and an airacetylene flame, using a 1:5 dilution of the Diluent as the blank. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of lead, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of lead in the Sample solution. Calculate the percentage of lead (Pb) in the portion of Bismuth Subcarbonate taken: Result = C/12,500 C = concentration of lead in the Sample solution LIMIT OF ALKALIES AND ALKALINE EARTHS Sample solution: Boil 1.0 g with 20 mL of a mixture of acetic acid and water (1:1). After 2 min, cool and filter. Analysis: Collect the filtrate, wash the residue with 20 mL of water, and add the washing to the filtrate. To this solution add 2 mL of 2 N hydrochloric acid and 20 mL of water. Heat to boiling and precipitate the bismuth by adding hydrogen sulfide. Cool the mixture, and filter.
Bismuth Subcarbonate
(Comment on this Monograph)id=m9760=Bismuth Subcarbonate=B-Monos.pdf) DEFINITION Bismuth Subcarbonate contains NLT 97.6% and NMT 100.7% of (BiO)2CO3, calculated on the dried basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Bismuth 191 and Carbonate 191 ASSAY PROCEDURE Sample solution: 500 mg of Bismuth Subcarbonate in 3 mL of nitric acid. Dilute with water to 250 mL, add 0.3 mL of xylenol orange TS Analysis: Titrate with 0.05 M edetate disodium VS to a yellow endpoint. Each mL of 0.05 M edetate disodium is equivalent to 12.75 mg of (BiO)2CO3. Acceptance criteria: 97.6%100.7% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221 Sample stock solution: 5.0 g in 10 mL of water. Add 20 mL of nitric acid, warm to achieve dissolution, allow to cool, and dilute with water to obtain 100 mL of solution. Sample solution: To 6.6 mL of the Sample stock solution add 4 mL of nitric acid, and dilute with water to 50 mL. Acceptance criteria: A 15.0-mL portion of the Sample solution shows no more chloride than corresponds to 70 L of 0.020 N hydrochloric acid (0.05%). LIMIT OF ARSENIC, Method I 211 Sample solution: 600 mg in 35 mL of 3 N hydrochloric acid Acceptance criteria: NMT 5 ppm LIMIT OF NITRATE Indigo carmine titrant: 4 g of indigo carmine in 900 mL of water, add 2 mL of sulfuric acid, and dilute with water to 1000 mL Standard solution: 0.0815 mg/mL of potassium nitrate (equivalent to 0.05 mg/mL of nitrate). Place 20.0 mL in a 125-mL conical flask. Sample solution: 250 mg of Bismuth Subcarbonate in a 125-mL conical flask, add 20 mL of water, and swirl to suspend
94

Collect the filtrate, wash the residue with water, and add the washing to the filtrate. Evaporate this solution to dryness on a water bath. To the residue add 0.5 mL of sulfuric acid, dry slowly, and cool. Acceptance criteria: The weight of the residue does not exceed 10 mg (1.0%).
USP 32
Acceptance criteria: No reddish brown color appears at the zone of contact of the two liquids. COPPER, LEAD, AND SILVER Sample: Ignite 3 g in a porcelain crucible, cool, and cautiously add, dropwise, just sufficient nitric acid to dissolve the residue upon warming. Evaporate the solution to dryness, again ignite, and cool. Cautiously dissolve the residue in just sufficient nitric acid with the aid of gentle heat, concentrate the solution to about 4 mL, pour it into 100 mL of water, filter, evaporate the filtrate on a steam bath to 20 mL, again filter, and divide this filtrate into portions of 5 mL each. Acceptance criteria Copper: Add a slight excess of 6 N ammonium hydroxide: the liquid does not exhibit a bluish color. Lead: Add 5 mL of 2 N sulfuric acid: the liquid does not become cloudy. Silver: Add hydrochloric acid, dropwise: no precipitate is formed that is insoluble in a slight excess of hydrochloric acid. LIMIT OF ALKALIES AND ALKALINE EARTHS Sample solution: Boil 1.0 g with 20 mL of a mixture of equal volumes of 6 N acetic acid and water, cool, and filter. Analysis: Precipitate the bismuth from the filtrate by the addition of hydrogen sulfide, boil the mixture, and filter. Add 5 drops of sulfuric acid to the filtrate, evaporate to dryness, and ignite to constant weight. Acceptance criteria: The weight of the residue does not exceed 5 mg (0.5%). Organic Impurities FREE GALLIC ACID Analysis: Shake 1.0 g with 20 mL of alcohol for 1 min, filter and evaporate the filtrate to dryness on a steam bath, and dry the residue at 105 for 1 h. Acceptance criteria: The weight of the residue does not exceed 5 mg (0.5%). SPECIFIC TESTS LOSS ON DRYING 731: 7.0% of its weight. Dry it at 105 for 3 h: it loses NMT
SPECIFIC TESTS LOSS ON DRYING 731: Dry at 105 to constant weight: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, protected from light.
Bismuth Subgallate
(Comment on this Monograph)id=m9770=Bismuth Subgallate=B-Monos.pdf)
C7H5BiO6 Gallic acid bismuth basic salt [99-26-3].
394.09
DEFINITION Bismuth Subgallate is a basic salt which, when dried at 105 for 3 h, contains the equivalent of NLT 52.0% and NMT 57.0% of Bi2O3. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Bismuth 191: Meets the requirements Sample: When heated to redness, it at first chars, leaving finally a yellow residue. B. PROCEDURE Analysis: Agitate thoroughly 100 mg with an excess of hydrogen sulfide TS, filter, and boil the filtrate to expel the dissolved gas. Cool, and add 1 drop of ferric chloride TS. Acceptance criteria: A purplish blue mixture is produced. ASSAY PROCEDURE Sample solution: Dry 1 g of Bismuth Subgallate at 105 for 3 h, then weigh accurately and ignite in a porcelain crucible. Allow it to cool and add nitric acid to the residue, dropwise, warming until complete solution has been effected. Analysis: Evaporate Sample solution to dryness and carefully ignite the residue to constant weight. From the weight of the residue so obtained, determine the percentage of Bi2O3 in the portion of Bismuth Subgallate taken. Acceptance criteria: 52.0%57.0% IMPURITIES Inorganic Impurities ARSENIC 211 Sample: Triturate 400 mg with an equal weight of calcium hydroxide, and ignite. Dissolve the residue in 5 mL of 3 N hydrochloric acid. Acceptance criteria: The solution, without further treatment, meets the requirements (NMT 7.5 ppm). LIMIT OF NITRATE Sample: Mix 100 mg with 5 mL of 2 N sulfuric acid and 5 mL of ferrous sulfate TS, filter the mixture, and carefully superimpose the filtrate, without mixing, on 5 mL of sulfuric acid, in a test tube.
Bismuth Subnitrate
(Comment on this Monograph)id=m9780=Bismuth Subnitrate=B-Monos.pdf) 1461.99 Bi5O(OH)9(NO3)4 Bismuth hydroxide nitrate oxide Bi5O(OH)9(NO3)4 [1304-85-4]. DEFINITION Bismuth Subnitrate is a basic salt that contains the equivalent of NLT 79.0% of bismuth trioxide (Bi2O3), calculated on the dried basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Bismuth 191 and Nitrate 191: Meets the requirements ASSAY PROCEDURE Sample solution: 400 mg of Bismuth Subnitrate to a 250mL beaker. Add 5 mL of water, then add 2 mL of nitric acid, and warm, if necessary, to effect solution. Dilute to 100 mL, and add 0.3 mL of xylenol orange TS.
USP 32
Analysis: Titrate with 0.05 M edetate disodium VS to a yellow endpoint. Each mL of 0.05 M edetate disodium is equivalent to 11.65 mg of Bi2O3. Acceptance criteria: NLT 79.0% of Bi2O3, calculated on the dried basis. IMPURITIES Inorganic Impurities CARBONATE: Add 3 g to 3 mL of warm nitric acid: no effervescence occurs. Pour the solution into 100 mL of water: a white precipitate forms. Filter, evaporate the filtrate on a steam bath to 30 mL, again filter the liquid, divide the latter filtrate into portions of 5 mL each, and use these several portions in the tests for Chloride, Sulfate, Copper, Lead, and Silver. CHLORIDE AND SULFATE, Chloride 221: A 10-mL portion of the test liquid retained in the test for Carbonate shows no more chloride than corresponds to 0.50 mL of 0.020 N hydrochloric acid (0.035%). CHLORIDE AND SULFATE, Sulfate 221: To a 5-mL portion of the test liquid retained in the test for Carbonate, add 5 drops of barium nitrate TS. Acceptance criteria: No turbidity is produced immediately. ARSENIC, Method I 211: Mix 375 mg with 5 mL of water, cautiously add 2 mL of sulfuric acid, and heat the mixture until fumes of sulfur trioxide are copiously evolved. Cool, cautiously add 10 mL of water, and again evaporate to strong fuming, repeating, if necessary, to remove any trace of nitric acid. Acceptance criteria: NMT 8 ppm COPPER: To a 5-mL portion of the test liquid retained in the test for Carbonate, add a slight excess of 6 N ammonium hydroxide. Acceptance criteria: The liquid does not exhibit a bluish color. LEAD: Mix a 5-mL portion of the test liquid retained in the test for Carbonate with an equal volume of 2 N sulfuric acid. Acceptance criteria: The liquid does not become cloudy. SILVER: To a 5-mL portion of the test liquid retained in the test for Carbonate, add hydrochloric acid, dropwise. Acceptance criteria: No precipitate is formed that is insoluble in a slight excess of hydrochloric acid, but that is soluble in 6 N ammonium hydroxide. LIMIT OF ALKALIES AND ALKALINE EARTHS: Boil 1.0 g with 20 mL of a mixture of equal volumes of 6 N acetic acid and water, cool, and filter. Add 2 mL of 3 N hydrochloric acid, precipitate the bismuth by the addition of hydrogen sulfide, boil the mixture, and filter it. Add 5 drops of sulfuric acid to the filtrate, evaporate to dryness, and ignite to constant weight. Acceptance criteria: The weight of the residue does not exceed 5 mg (0.5%). LIMIT OF AMMONIUM SALTS: Boil about 100 mg with 5 mL of 1 N sodium hydroxide. Acceptance criteria: The vapor does not turn moistened red litmus paper blue. SPECIFIC TESTS LOSS ON DRYING 731: 3.0% of its weight. Dry it at 105 for 2 h: it loses NMT

Bismuth Subsalicylate
(Comment on this Monograph)id=m9785=Bismuth Subsalicylate=B-Monos.pdf) C7H5BiO4 362.09 (2-Hydroxybenzoato-O1)-oxobismuth; 2-Hydroxybenzoic acid bismuth (3+) salt, basic [14882-18-9]. DEFINITION Bismuth Subsalicylate is a basic salt that when dried at 105 for 3 h contains NLT 56.0% and NMT 59.4% of bismuth (Bi) and NLT 36.5% and NMT 39.3% of total salicylates. IDENTIFICATION A. INFRARED ABSORPTION 197M B. IDENTIFICATION TESTSGENERAL, Bismuth 191: requirements
Meets the
ASSAY BISMUTH Sample solution: 300 mg of Bismuth Subsalicylate, previously dried at 105 for 3 h in a porcelain crucible Ignite, allow it to cool, and add about 2 mL of nitric acid to the residue, dropwise, warming until complete solution has been effected. Add about 60 mL of water and 0.3 mL of xylenol orange TS. Analysis: Titrate with 0.05 M edetate disodium VS to a yellow endpoint. Each mL of 0.05 M edetate disodium is equivalent to 10.45 mg of Bi. Acceptance criteria: 56.0%59.4% TOTAL SALICYLATES Ferric ammonium sulfate solution: Ferric ammonium sulfate TS, 1 N hydrochloric acid, and water (4:1:15) Standard stock solution: 0.2 mg/mL of USP Salicylic Acid RS in water Standard solution: 0.05 mg/mL salicylic acid in water [NOTEAdjust with 0.5 N sodium hydroxide or 1 N hydrochloric acid to a pH of 4.5, prior to final dilution.] Sample solution: 52 mg of Bismuth Subsalicylate, previously dried at 105 for 3 h, in a 200-mL volumetric flask Add 10 mL of 0.5 N sodium hydroxide, heat on a steam bath for 15 min, allow to cool, and dilute with water to volume. Centrifuge about 70 mL of this solution, then transfer 50.0 mL of the clear supernatant to a beaker. Add about 40 mL of water, and adjust with 0.5 N sodium hydroxide or 1 N hydrochloric acid to a pH of 4.5. Transfer this solution to a 100-mL volumetric flask with the aid of water, and dilute with water to volume. Blank: Water, adjusted with 0.5 N sodium hydroxide or 1 N hydrochloric acid to a pH of 4.5 Analysis Samples: Standard solution, Sample solution, and Blank Prepare the following solutions in separate 50-mL conical flasks.
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Reacted standard solution: To 25.0 mL of Standard solution, add 1.0 mL of Ferric ammonium sulfate solution. Reacted sample solution: To 25.0 mL of Sample solution, add 1.0 mL of Ferric ammonium sulfate solution. Reacted blank solution: To 25.0 mL of Blank, add 1.0 mL of Ferric ammonium sulfate solution. Unreacted standard solution: To 25.0 mL of the Standard solution, add 1.0 mL of 0.05 N hydrochloric acid. Unreacted sample solution: To 25.0 mL of the Sample solution, add 1.0 mL of 0.05 N hydrochloric acid. Unreacted blank solution: To 25.0 mL of Blank, add 1.0 mL of 0.05 N hydrochloric acid. Concomitantly determine the absorbances of these six solutions at the wavelength of maximum absorption at about 525 nm, using water to zero the spectrophotometer. Calculate the percentage of total salicylates in the portion of C7H5BiO4 taken: Result = [(AUR AUU B)/(ASR ASU B)] (CS/W) 10,000
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Scattering 851) equipped with copper, lead, and silver hollow-cathode lamps and an oxidizing flame. Acceptance criteria: NMT 10 ppm LIMIT OF SOLUBLE BISMUTH Standard solution: 242.0 mg of bismuth nitrate pentahydrate in a 100-mL volumetric flask Add 3 mL of 1.5 M nitric acid, swirl to dissolve, and dilute with water to volume. Transfer 1.0 mL of this solution to a 500-mL volumetric flask, add 250 mL of 1.5 M nitric acid, dilute with water to volume, and mix. This solution contains 2 g/mL of bismuth. [NOTEThe concentration of bismuth in this solution may be modified by using a lesser dilution or by further dilution to bring the absorption response within the working range of the atomic absorption spectrophotometer.] Sample solution: 5.0 g of Bismuth Subsalicylate in 100 mL of water Stir the suspension thus obtained for 2 h at 2023. Filter through filter paper. Filter the filtrate thus obtained through a filter having a porosity of 0.1 m or less. To 10.0 mL of the filtrate add 0.1 mL of nitric acid. [NOTEThe concentrate of Bismuth Subsalicyclate may be modified by using the same proportions used for modifying the Standard Solution, by using a different quantity, or by further dilution.] Analysis Samples: Standard solution and Sample solution Concomitantly determine the absorbances of the Standard solution and the Sample solution at the emission line of 223.06 nm for bismuth with an atomic absorption spectrophotometer (see Spectrophotometry and Light-Scattering 851) equipped with a bismuth hollow-cathode lamp and an oxidizing flame. Acceptance criteria: NMT 40 ppm LIMIT OF NITRATE Sample solution: To 0.1 g add 10 mL of water, and carefully add 20 mL of sulfuric acid. Standard solution: Add to 0.1 g of salicylic acid, 6 mL of water, 4.0 mL of a solution containing 100 g of nitrate per mL, and 20 mL of sulfuric acid. Acceptance criteria: The Sample solution should not be more yellow than the Standard solution (0.4%), prepared concomitantly. Organic Impurities LIMIT OF FREE SALICYLIC ACID Mobile phase: Methanol and 0.06 M acetic acid (11:9) Diluent: Acetonitrile and water (1:1) Standard solution: 0.02 mg/mL of USP Salicylic Acid RS in Diluent Sample solution: 260 mg of Bismuth Subsalicylate in a glass centrifuge tube Add about 12 mL of acetonitrile, shake by mechanical means for 20 min, and centrifuge. Decant the supernatant into a suitable container. Repeat the acetonitrile addition, shaking, centrifuging, and decanting, combining the decanted liquid with the first decantate. Pass the combined liquid through a filter having a 0.5-m or finer porosity, collecting the filtrate in a 50-mL volumetric flask. Wash the container with 5 mL of acetonitrile, and filter the wash, collecting the filtrate in the volumetric flask. Dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.)
= absorbance of the Reacted sample solution = absorbance of the Unreacted sample solution = difference in the absorption of the Reacted blank solution and the absorption of the Unreacted blank solution = absorbance of the Reacted standard solution ASR = absorbance of the Unreacted standard solution ASU = concentration of USP Salicylic Acid RS in the CS Standard stock solution (mg/mL) W = weight, in mg, of Bismuth Subsalicylate taken Acceptance criteria: 36.5%39.3% AUR AUU B IMPURITIES Inorganic Impurities ARSENIC, Method I 211: Triturate 300 mg of bismuth subsalicylate with 300 mg of calcium hydroxide, and ignite. Dissolve the residue in 5 mL of 3 N hydrochloric acid. Acceptance criteria: NMT 10 ppm LIMIT OF COPPER, LEAD, AND SILVER Standard solution: 3.0 mL each of solutions containing 1000 g/mL of copper, lead, and silver, respectively, to a 2000-mL volumetric flask Dilute with 1 M nitric acid to volume. [NOTEThe concentrations of copper, lead, and silver may be modified by using different volumes or concentrations to bring the absorption response within the working range of the atomic absorption spectrophotometer.] Sample solution: Ignite 3 g of sample in a porcelain crucible, cool, cautiously add 6 M nitric acid to dissolve the residue, and evaporate on a steam bath. Ignite the residue, cool, and transfer the residue to a tared conical flask. Wash the flask with about 5 mL of 6 M nitric acid, adding the wash to the conical flask. Dissolve the residue with the aid of heat, and add water to obtain a solution weighing 20.0 g. [NOTEThe concentrate of Bismuth Subsalicyclate may be modified by using the same proportions used for modifying the Standard solution, by using a different quantity, or by further dilution.] Analysis Samples: Standard solution and Sample solution Concomitantly determine the absorbances of the Standard solution and the Sample solution at the emission lines of 324.7 nm, 217 nm, and 328.1 nm for copper, lead, and silver, respectively, with an atomic absorption spectrophotometer (see Spectrophotometry and Light-
USP 32
Mode: LC Detector: UV 300 nm Column: 4.6-mm 30-cm; 5-m packing L1 Guard column: 3.2-mm 1.5-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of free salicylic acid in C7H5BiO4: Result = (rU/rS) (CS/CU) 100 = peak area response of salicylic acid in the Sample solution = peak area response of the Standard solution rS = concentration of USP Salicylic Acid RS in the CS Standard solution (mg/mL) = concentration of the Bismuth Subsalicylate in CU the Sample solution (mg/mL) Acceptance criteria: NMT 0.2% rU SPECIFIC TESTS PH 791 Analysis: Place 10 g of Bismuth Subsalicylate in 90 mL of water, shake by mechanical means for 10 min, and filter. Acceptance criteria: 2.75.0 LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Bismuth Subsalicylate RS USP Salicylic Acid RS

Acceptance criteria: 56.0%59.4% of Bi TOTAL SALICYLATES Solution A: Ferric ammonium sulfate TS, 1 N hydrochloric acid, and water (4:1:15) Standard stock solution: 0.2 mg/mL of USP Salicylic Acid RS in water Standard solution: 0.05 mg/mL salicylic acid in water, adjust with 0.5 N sodium hydroxide or 1 N hydrochloric acid to a pH of 4.5, prior to final dilution Reacted standard solution: To 25.0 mL of Standard solution, add 1.0 mL of Solution A. Unreacted standard solution: To 25.0 mL of the Standard solution, add 1.0 mL of 0.05 N hydrochloric acid. Sample solution: Equivalent to 52 mg of bismuth subsalicylate from dried magma in a 200-mL volumetric flask. Add 10 mL of 0.5 N sodium hydroxide, heat on a steam bath for 15 min, allow to cool, and dilute with water to volume. Centrifuge and then transfer 50.0 mL of the clear supernatant to a beaker. Add about 40 mL of water, and adjust with 0.5 N sodium hydroxide or 1 N hydrochloric acid to a pH of 4.5. Transfer this solution to a 100-mL volumetric flask with the aid of water, and dilute with water to volume. Reacted sample solution: To 25.0 mL of Sample solution, add 1.0 mL of Solution A. Unreacted sample solution: To 25.0 mL of the Sample solution, add 1.0 mL of 0.05 N hydrochloric acid. Blank: Water, adjusted with 0.5 N sodium hydroxide or 1 N hydrochloric acid to a pH of 4.5 Reacted blank solution: To 25.0 mL of Blank, add 1.0 mL of Solution A. Unreacted blank: To 25.0 mL of Blank, add 1.0 mL of 0.05 N hydrochloric acid. Analysis Samples: Reacted standard solution, Unreacted standard solution, Reacted sample solution, Unreacted sample solution, Reacted blank solution, and Unreacted blank Concomitantly determine the absorbances of the Samples at the wavelength of maximum absorption at about 525 nm, using water to zero the spectrophotometer. Calculate the percentage of total salicylates in the portion of dried magma taken: Result = [(AUR AUU B)/(ASR ASU B)] (CS/W) 10,000 = absorbance of the Reacted sample solution = absorbance of the Unreacted sample solution = difference in the absorption of the Reacted blank solution and the absorption of the Unreacted blank ASR = absorbance of the Reacted standard solution ASU = absorbance of the Unreacted standard solution CS = concentration of USP Salicylic Acid RS in the Standard stock solution (mg/mL) W = weight, in mg, of Bismuth Subsalicylate taken for the Sample solution Acceptance criteria: 36.5%39.3% of total salicylates AUR AUU B IMPURITIES Inorganic Impurities LIMIT OF COPPER, LEAD, AND SILVER Standard solution: 1.5 g/mL of copper, 1.5 g/mL of lead, and 1.5 g/mL of silver, in 1 M nitric acid [NOTEThe concentrations of copper, lead, and silver may be modified by using different volumes or concentrations to bring the absorption response within the working range of the atomic absorption spectrophotometer.] Sample solution: Ignite 3 g of sample in a porcelain crucible, cool, and cautiously add 6 M nitric acid to dissolve the residue, and evaporate on a steam bath. Ignite the residue, cool, and transfer the residue to a tared conical flask, and wash the flask with about 5 mL of 6 M nitric
Bismuth Subsalicylate Magma

(Comment on this Monograph)id=m9786=Bismuth Subsalicylate Magma=B-Monos.pdf) DEFINITION Bismuth Subsalicylate Magma is a suspension of Bismuth Subsalicylate in water that contains NLT 90.0% and NMT 110.0% of the labeled amount of C7H5O4Bi. Bismuth Subsalicylate is a basic salt that when dried at 105 for 3 h contains NLT 56.0% and NMT 59.4% bismuth and NLT 36.5% and NMT 39.3% of total salicylates. [NOTEDry at 105 for 3 h to determine the solids content and, after determining the solids content, perform all tests on a portion of the dried magma.] IDENTIFICATION A. INFRARED ABSORPTION 197M B. IDENTIFICATION TESTSGENERAL, Bismuth 191: requirements
Meets the
ASSAY BISMUTH Sample solution: Ignite an equivalent to 300 mg of bismuth subsalicylate from dried magma. Allow it to cool, and add about 2 mL of nitric acid to the residue, dropwise, warming until complete solution has been effected. Add about 60 mL of water and 0.3 mL of xylenol orange TS. Analysis: Titrate the Sample solution with 0.05 M edetate disodium VS to a yellow endpoint. Each mL of 0.05 M edetate disodium is equivalent to 10.45 mg of bismuth (Bi).
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USP 32
decanting, combining the decanted liquid with the first decantate. Pass the combined liquid through a filter having a 0.5-m or finer porosity, collecting the filtrate in a 50-mL volumetric flask. Wash the container with 5 mL of acetonitrile, and filter the wash, collecting the filtrate in the volumetric flask. Dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 300 nm Guard column: 3.2-mm 1.5-cm; 5-m packing L1 Analytical column: 4.6-mm 30-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of free salicylic acid in the Bismuth Subsalicylate taken: Result = (rU/rS) (CS/CU) 100 = peak area of salicylic acid in the Sample solution = peak area of the Standard solution = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) = concentration of the bismuth subsalicylate in CU the Sample solution (mg/mL) Acceptance criteria: NMT 0.2% rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: The label states that this article is not intended for direct administration to humans or animals. USP REFERENCE STANDARDS 11 USP Bismuth Subsalicylate RS USP Salicylic Acid RS
acid, adding the wash to the conical flask. Dissolve the residue with the aid of heat, and add water to obtain a solution weighing 20.0 g. [NOTEThe concentrate of bismuth subsalicyclate may be modified by using the same proportions used for modifying the Standard solution, by using a different quantity, or by further dilution.] Analysis Samples: Standard solution and Sample solution Concomitantly determine the absorbances of the Standard solution and the Sample solution at the emission lines of 324.7 nm, 217 nm, and 328.1 nm, for copper, lead, and silver, respectively, with an atomic absorption spectrophotometer (see Spectrophotometry and Lightscattering 851) equipped with copper, lead, and silver hollow-cathode lamps and an oxidizing flame. Acceptance criteria: The absorbances of the Sample solution do not exceed those of the Standard solution for each element (NMT 10 ppm). LIMIT OF SOLUBLE BISMUTH Standard solution: 242.0 mg of bismuth nitrate pentahydrate in a 100-mL volumetric flask, add 3 mL of 1.5 M nitric acid and swirl to dissolve, dilute with water to volume, and mix. Transfer 1.0 mL of this solution to a 500mL volumetric flask, add 250 mL of 1.5 M nitric acid, dilute with water to volume, and mix (2 g/mL of bismuth). [NOTEThe concentration of bismuth in this solution may be modified by using a lesser dilution or by further dilution to bring the absorption response within the working range of the atomic absorption spectrophotometer.] Sample solution: 5.0 g of bismuth subsalicylate in 100 mL of water, and stir the suspension thus obtained for 2 h at 2023. Pass through filter paper. Pass the filtrate thus obtained through a filter having a porosity of 0.1 m or less. To 10.0 mL of the filtrate add 0.1 mL of nitric acid. [NOTEThe concentrate of bismuth subsalicyclate may be modified by using the same proportions used for modifying the Standard solution, by using a different quantity, or by further dilution.] Analysis Samples: Standard solution and Sample solution Concomitantly determine the absorbances of the Standard solution and the Sample solution at the emission line of 223.06 nm for bismuth with an atomic absorption spectrophotometer (see Spectrophotometry and Lightscattering 851) equipped with a bismuth hollowcathode lamp and an oxidizing flame. Acceptance criteria: The absorbances of the Sample solution do not exceed those of the Standard solution (NMT 40 ppm). LIMIT OF NITRATE Sample solution: To 0.1 g add 10 mL of water, carefully add 20 mL of sulfuric acid, and mix. Standard solution: Add to 0.1 g of salicylic acid, 6 mL of water, 4.0 mL of a solution containing 100 g of nitrate (NO3) per mL, and 20 mL of sulfuric acid Acceptance criteria: The Sample solution should not be more yellow than the Standard solution (0.4%), prepared concomitantly. Organic Impurities LIMIT OF FREE SALICYLIC ACID Mobile phase: Methanol and 0.06 M acetic acid (11:9) Diluent: Acetonitrile and water (1:1) Standard solution: 0.02 mg/mL of USP Salicylic Acid RS in Diluent Sample solution: 260 mg of bismuth subsalicylate, in a glass centrifuge tube, add about 12 mL of acetonitrile, shake by mechanical means for 20 min, and centrifuge. Decant the supernatant into a suitable container. Repeat the acetonitrile addition, shaking, centrifuging, and
Bismuth Subsalicylate Oral Suspension

(Comment on this Monograph)id=m9787=Bismuth Subsalicylate Oral Suspension=B-Monos.pdf) DEFINITION Bismuth Subsalicylate Oral Suspension is a suspension that contains NLT 90.0% and NMT 110.0% of the labeled amount of C7H5BiO4. It may contain one or more suitable buffers, coloring agents, flavors, preservatives, stabilizers, sweeteners, and suspending agents. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Bismuth 191 B. IDENTIFICATION TESTSGENERAL, Salicylate 191: acidifying with nitric acid
After
ASSAY PROCEDURE Standard stock solution: 500 mg of bismuth metal, in a 200-mL volumetric flask, dissolve in 12 mL of nitric acid, and dilute with 0.01 N nitric acid to volume Standard solution: 50 g/mL from Standard stock solution in 1 N nitric acid Sample solution: 10 g of Oral Suspension, previously wellshaken in its original container to ensure homogeneity, in a 200-mL volumetric flask. Add about 100 mL of 1 N nitric acid, mix, and dilute with 1 N nitric acid to volume. Mix
USP 32
well without shaking, transfer 10.0 mL of this mixture to a 100-mL volumetric flask, and dilute with 1 N nitric acid to volume. Centrifuge about 20 mL at 4500 rpm for at least 10 min. Spectrometric conditions Mode: UV-Vis Analytical wavelength: 463 nm Cell: 1 cm Analysis Samples: Standard solution and Sample solution Transfer an accurately measured volume of the Sample solution that contains 0.9 mg of bismuth subsalicylate and 10 mL of the Standard solution to separate 50-mL volumetric flasks. Add 10.0 mL of 10% ascorbic acid solution and 25.0 mL of 20% potassium iodide solution into each volumetric flask, dilute with water to volume, and mix well. Concomitantly determine the absorbances of both solution, using the reagent blank to set the spectrophotometer. Calculate the percentage of bismuth subsalicylate (C7H5BiO4) in the portion of Oral Suspension: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of bismuth in the Standard solution (g/mL) = nominal concentration of the Sample solution CU (g/mL) = molecular weight of bismuth subsalicylate, Mr1 362.09 = molecular weight of bismuth, 208.98 Mr2 Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62 Total aerobic microbial count: NMT 100 cfu/g Total combined yeasts and molds count: NMT 50 cfu/g It meets the requirements of the tests for absence of Escherichia coli. PH 791: 3.05.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Protect from freezing. Avoid excessive heat (over 40). AU AS CS
Official Monographs / Bisoctrizole 99

Sample stock solution: Equivalent to 90 mg of bismuth subsalicylate from powdered Tablets, in a 200-mL volumetric flask, add 150 mL of 1 N nitric acid, and sonicate for 2 min. Dilute with 1 N nitric acid to volume. Sample solution: Sample stock solution and 1 N nitric acid (1:4). Centrifuge a portion at 4500 rpm for at least 10 min. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 463 nm Cell: 1 cm Analysis Samples: Standard solution and Sample solution Analysis: Transfer 10.0 mL of the Sample solution and the Standard solution to separate 50.0-mL volumetric flasks. Add 10.0 mL of 10% ascorbic acid solution and 25.0 mL of 20% potassium iodide solution into each volumetric flask, and dilute with 1 N nitric acid to volume. Concomitantly determine the absorbance of the solutions at the wavelength of maximum absorbance at 463 nm with a suitable spectrophotometer using the combined reagent solutions as the blank. Calculate the percentage of C7H5BiO4 in the portion of Tablets: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of bismuth in the Standard solution (g/mL) CU = nominal concentration of bismuth in the Sample solution (g/mL) Mr1 = molecular weight of bismuth subsalicylate, 362.09 = molecular weight of bismuth, 208.98 Mr2 Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701 Time: 10 min [NOTEThis test does not apply for Tablets labeled as chewable.] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Avoid excessive heat (over 40). LABELING: Label chewable Tablets to indicate that they are to be chewed before swallowing. AU AS CS
Bismuth Subsalicylate Tablets

(Comment on this Monograph)id=m9788=Bismuth Subsalicylate Tablets=B-Monos.pdf) DEFINITION Bismuth Subsalicylate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of bismuth subsalicylate (C7H5BiO4). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Bismuth 191 B. IDENTIFICATION TESTSGENERAL, Salicylate 191: After acidifying with nitric acid, it meets the requirements of the test with ferric chloride TS. ASSAY PROCEDURE Standard stock solution: 500 mg of bismuth, in a 200-mL volumetric flask, dissolve in 12 mL of nitric acid, and dilute with 0.01 N nitric acid to volume Standard solution: 50 g/mL of bismuth from Standard stock solution and 1 N nitric acid (1:49)
Bisoctrizole
(Comment on this Monograph)id=m1129=Bisoctrizole=BMonos.pdf)
C41H50N6O2 658.90 Phenol, 2,2-methylenebis[6-(2H-benzotriazol-2-yl)-4-(1,1,3,3tetramethylbutyl)]-; 2,2-Methylenebis[6-(2H-benzotriazol-2-yl)-4-(1,1,3,3tetramethylbutyl)phenol] [103597-45-1]. DEFINITION Bisoctrizole contains NLT 96.0% and NMT 102.0% of C41H50N6O2, calculated on the as-is basis.
100
Bisoctrizole / Official Monographs

USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Tetrahydrofuran and a 0.2% (w/v) aqueous solution of 1-pentane sulfonic acid sodium salt (3:2) Solution A: 0.4 g of 1-pentane sulfonic acid sodium salt, 800 mL of methanol, 200 mL of water, and 0.5 mL of phosphoric acid Solution B: 0.4 g of 1-pentane sulfonic acid sodium salt, 1000 mL of methanol, and 0.5 mL of phosphoric acid Mobile phase: See gradient table below.
IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1 Diluent, Solution A, Solution B, Mobile phase, Standard solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Bisoctrizole taken: Result = (ru/rT) 100 ru = peak response for each individual impurity rT = sum of the responses of all the peaks Acceptance criteria Individual impurities: NMT 0.1% of any individual impurity, excluding bisoctrizole related compound A and the bisoctrizole isomer Total impurities: NMT 4.0% of total impurities, including bisoctrizole related compound A, and the bisoctrizole isomer, determined in the test for Limit of bisoctrizole related compound A and the bisoctrizole isomer PROCEDURE 2: LIMIT OF BISOCTRIZOLE RELATED COMPOUND A AND THE BISOCTRIZOLE ISOMER Diluent, Solution A, Solution B, Mobile phase, Sample solution, and Chromatographic system: Proceed as directed in the Assay. Standard stock solution A: 0.65 mg/mL of USP Bisoctrizole RS in tetrahydrofuran Standard stock solution B: 0.40 mg/mL of USP Bisoctrizole Related Compound A RS in tetrahydrofuran Standard solution: Standard stock solution A, Standard stock solution B, tetrahydrofuran, and Diluent (5:1:60:34) System suitability Sample: Standard solution [NOTEThe relative retention times for bisoctrizole related compound A and the bisoctrizole isomer are about 0.42 and about 1.1, respectively.] Suitability requirements Resolution: NLT 1.5 between bisoctrizole and the bisoctrizole isomer Analysis Samples: Standard solution and Sample solution Calculate the percentage of bisoctrizole related compound A taken: Result = (rU/rS) (CS/CU) 100 = peak response for the bisoctrizole related compound A in the Sample solution = peak response for bisoctrizole related compound rS A in the Standard solution CS = concentration of USP Bisoctrizole Related Compound A RS in the Standard solution (mg/mL) = nominal concentration of bisoctrizole related CU compound A in the Sample solution (mg/mL) Acceptance criteria: NMT 0.5% of the bisoctrizole related compound A rU
System suitability solution: 0.8 mg/mL of bisoctrizole from USP Bisoctrizole Resolution Mixture RS dissolved in tetrahydrofuran, and dilute with Diluent Standard solution: 80 mg of USP Bisoctrizole RS to a 100mL volumetric flask. Dissolve in 60 mL of tetrahydrofuran, and dilute with Diluent to volume. Sample solution: 80 mg of Bisoctrizole to a 100-mL volumetric flask. Dissolve in 60 mL of tetrahydrofuran, and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 346 nm Column: 3.0-mm 25-cm; packing L1 Temperature: 40 Flow rate: 0.8 mL/min Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for bisoctrizole and the bisoctrizole isomer are about 1.0 and 1.1, respectively.] Suitability requirements Resolution: NLT 1.5 between bisoctrizole and the bisoctrizole isomer, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C41H50N6O2 in the portion of Bisoctrizole taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bisoctrizole RS in the Standard solution (mg/mL) = nominal concentration of bisoctrizole in the Sample solution (mg/mL)
USP 32
Calculate the percentage of bisoctrizole isomer taken: Result = (rU/rS) (CS/CU) 100 = peak response for the bisoctrizole isomer in the Sample solution = peak response for Bisoctrizole in the Standard rS solution = concentration of USP Bisoctrizole RS in the CS Standard solution (mg/mL) = nominal concentration of bisoctrizole in the CU Sample solution (mg/mL) Acceptance criteria: NMT 4.0% of the bisoctrizole isomer rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Bisoctrizole RS USP Bisoctrizole Related Compound A RS USP Bisoctrizole Resolution Mixture RS
Official Monographs / Bisoprolol 101

Sample solution: 1 mg/mL of Bisoprolol Fumarate in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 273 nm Column: 4.6-mm 12.5-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 7.0 between bisoprolol and propranolol in the System suitability solution Tailing factor: NMT 2.0 in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C18H31NO4)2 C4H4O4 in the portion of Bisoprolol Fumarate taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bisoprolol Fumarate RS in the Standard solution (mg/mL) = concentration of bisoprolol fumarate in the CU Sample solution (mg/mL) Acceptance criteria: 97.5%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method I 231: NMT 20 ppm Organic Impurities PROCEDURE Diluent, Mobile phase, System suitability solution, Standard solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay. System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 7.0 between bisoprolol and propranolol in the System suitability solution Tailing factor: NMT 2.0 in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Sample: Sample solution Calculate the percentage of total impurities in the portion of Bisoprolol Fumarate taken: Result = (ru/rT) 100 Ru RT = sum of areas for all the peaks, excluding the fumaric acid and bisoprolol peaks = sum of the areas of all the peaks in the chromatogram rU rS CS
Bisoprolol Fumarate
(Comment on this Monograph)id=m9792=Bisoprolol Fumarate=B-Monos.pdf)
766.96 (C18H31NO4)2 C4H4O4 2-Propanol, 1-[4-[[2-(1-methylethoxy)ethoxy]methyl] phenoxy]-3-[(1-methylethyl)amino]-, ()-, (E)-2-butenedioate (2:1) (salt); ()-1-[[-(2-Isoproproxyethoxy)-p-tolyl]oxy]-3(isopropylamino)-2-propanol fumarate (2:1) (salt) [104344-23-2]. DEFINITION Bisoprolol Fumarate contains NLT 97.5% and NMT 102.0% of (C18H31NO4)2 C4H4O4, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. LIQUID CHROMATOGRAPHY: The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Acetonitrile and water (7:13) Mobile phase: To a 1-L portion of Diluent add 5 mL of heptafluorobutyric acid, 5 mL of diethylamine, and 2.5 mL of formic acid. System suitability solution: 0.5 mg/mL of propranolol hydrochloride and 1 mg/mL of Bisoprolol Fumarate in Diluent Standard solution: 1 mg/mL of USP Bisoprolol Fumarate RS in Diluent
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Bisoprolol / Official Monographs
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Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 273 nm Column: 4.6-mm 12.5-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 7.0 between bisoprolol and propranolol, System suitability solution Tailing factor: NMT 2.0 in the Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C18H31NO4)2 C4H4O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bisoprolol Fumarate RS in the Standard solution (mg/mL) = nominal concentration of bisoprolol fumarate in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%105.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 20 min Analysis: Determine the amount of (C18H31NO4)2 C4H4O4 dissolved by employing the following method. Diluent: Methanol, triethylamine, phosphoric acid, and water (160:5:2.5:35) Mobile phase: Methanol, triethylamine, and water (34:1:50), adjust with phosphoric acid to a pH of 4.0 0.1. Standard stock solution: USP Bisoprolol Fumarate RS in water to obtain a solution having a known concentration of about twice the concentration of bisoprolol fumarate in the Sample solution Standard solution: Standard stock solution and Diluent (1:1) Sample solution: Sample per Dissolution 711. Withdraw a portion of the solution under test, filter, and dilute with an equal volume of Diluent. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 227 nm Column: 4.6-mm 33-cm; packing L7 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C18H31NO4)2 C4H4O4 dissolved. Tolerances: NLT 80% (Q) of the labeled amount of (C18H31NO4)2 C4H4O4 is dissolved. Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. rU rS CS
Acceptance criteria Total impurities: NMT 0.5% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781 Sample solution: 10 mg/mL, in methanol Acceptance criteria: 2 to +2 WATER DETERMINATION, Method I 921: NMT 0.5% CONTENT OF FUMARIC ACID Sample solution: 500 mg of Bisoprolol Fumarate in 70 mL of dehydrated alcohol Analysis: Add 8.0 mL of 0.1 N tetrabutylammonium hydroxide VS, and stir for 2 min. Titrate with 0.1 N tetrabutylammonium hydroxide VS, using a glass-calomel electrode system. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N tetrabutylammonium hydroxide is equivalent to 5.804 mg of fumaric acid. Acceptance criteria: NLT 14.8% and NMT 15.4% of fumaric acid is found, calculated on the anhydrous basis. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Bisoprolol Fumarate RS
Bisoprolol Fumarate Tablets

(Comment on this Monograph)id=m9794=Bisoprolol Fumarate Tablets=B-Monos.pdf) DEFINITION Bisoprolol Fumarate Tablets contain NLT 90.0% and NMT 105.0% of the labeled amount of bisoprolol fumarate [(C18H31NO4)2 C4H4O4]. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Equivalent to 40 mg of bisoprolol fumarate, from powdered Tablets (NLT 5), in a 50-mL flask. Add about 20 mL of a mixture of dichloromethane and methanol (7:3), shake for 30 min, centrifuge, and use the clear solution. Application volume: 20 L Developing solvent system: Dichloromethane, methanol, and ammonia TS, stronger (70:10:0.8) Analysis Sample: Sample solution Proceed as directed in the chapter, except to develop the chromatogram until the solvent front has moved about two-thirds of the length of the plate and to dry the plate in a current of cold air. ASSAY PROCEDURE Diluent: Acetonitrile and water (7:13) Mobile phase: 1-L portion of Diluent, add 5 mL of heptafluorobutyric acid, 5 mL of diethylamine, and 2.5 mL of formic acid System suitability solution: 0.5 mg/mL of propranolol hydrochloride and 1 mg/mL of bisoprolol fumarate in Diluent Standard solution: 1 mg/mL of USP Bisoprolol Fumarate RS in Diluent Sample solution: Transfer an equivalent of 25 mg of bisoprolol fumarate, from powdered Tablets (NLT 20), to a 25-mL volumetric flask. Add 10 mL of Diluent, and sonicate for 10 min. Cool, dilute with Diluent to volume, and mix. Centrifuge for 20 min, and use the clear supernatant.
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Medium: 0.5 M sodium chloride; 900 mL Apparatus 2: 75 rpm Time: 20 min Analysis: Proceed as directed in Performance Tests, Test 1, Analysis. Tolerances: NLT 80% (Q) of the labeled amount of (C18H31NO4)2 C4H4O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. LABELING: When more than one Dissolution test is given, the labeling states the Dissolution test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Bisoprolol Fumarate RS Mobile phase:
Time (min) 0 9.0 9.1 12.0
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See the gradient table below
Solution A (%) 100 40 100 100 Solution B (%) 0 60 0 0
Bisoprolol Fumarate and Hydrochlorothiazide Tablets

(Comment on this Monograph)id=m9796=Bisoprolol Fumarate and Hydrochlorothiazide Tablets=B-Monos.pdf) DEFINITION Bisoprolol Fumarate and Hydrochlorothiazide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of bisoprolol fumarate [(C18H31NO4)2 C4H4O4] and hydrochlorothiazide (C7H8ClN3O4S2). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution A: 1 mg/mL of USP Bisoprolol Fumarate RS in methanol Standard solution B: 1 mg/mL of USP Hydrochlorothiazide RS in methanol Sample solution: Finely powder 1 Tablet, and transfer the powder to a 5-mL volumetric flask. Dilute with methanol to volume, sonicate for 5 min, centrifuge, and use the supernatant. Application volume: 25 L Developing solvent system: Methylene chloride, methanol, and 14.5 M ammonium hydroxide solution (43:20:8) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Locate the spots on the plate under short-wavelength UV light and by exposure to iodine vapors. Acceptance criteria: The RF values of the principal spots of the Sample solution correspond to the principal spots of Standard solution A and Standard solution B. B. LIQUID CHROMATOGRAPHY: The retention times of the major peaks of the Sample solution A and Sample solution B corresponds to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: 10 mL of 1 M dibutylammonium phosphate per L of acetronitrile and water (1:1) Solution A: 1 M dibutylammonium phosphate and water (1:100) Solution B: 10 mL of 1 M dibutylammonium phosphate per L of acetronitrile and water (3:2), stir vigorously for 2 min, filter
System suitability solution: 40 g/mL of USP Chlorothiazide RS and 40 g/mL of USP Hydrochlorothiazide RS in Diluent Standard solution: 100 g/mL of USP Bisoprolol Fumarate RS and 100 g/mL of USP Hydrochlorothiazide RS in Diluent. Stir by mechanical means for 1 h. Sample stock solution: Weigh 10 Tablets, and transfer to a 100-mL volumetric flask. Add about 50 mL of Diluent, sonicate for 10 min, and cool. Dilute with Diluent to volume, stir by mechanical means for 1 h, and centrifuge. Sample solution A: 100 g/mL of bisoprolol fumarate from Sample stock solution in Diluent Sample solution B: 62.5 g/mL of hydrochlorothiazide from Sample stock solution in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 225 nm Column: 8-mm 10-cm; packing L11 Flow rate: 3 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.5 between chlorothiazide and hydrochlorothiazide in the System suitability solution Tailing factor: NMT 1.3 for the hydrochlorothiazide peak in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution, Sample solution A, and Sample solution B Calculate the percentage of (C18H31NO4)2 C4H4O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response for bisoprolol fumarate in Sample solution A = peak response for bisoprolol fumarate in the rS Standard solution = concentration of USP Bisoprolol Fumarate RS in CS the Standard solution (mg/mL) = nominal concentration of bisoprolol fumarate in CU Sample solution A (mg/mL) Calculate the percentage of C7H8ClN3O4S2 in the portion of Tablets taken: rU Result = (rU/rS) (CS/CU) 100 rU rS = peak response for hydrochlorothiazide in Sample solution B = peak response for hydrochlorothiazide in the Standard solution
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CS
Bisoprolol / Official Monographs
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Tailing factor: NMT 1.3 in Standard solution Relative standard deviation: NMT 2.0% in Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Tablets taken: Result = (ru/rS) (WB/WH) (CS/CU) 1/F 100 = peak response of each of the two impurities from the Sample solution = peak response for hydrochlorothiazide in the rS Standard solution = labeled quantity of bisoprolol fumarate in each WB Tablet (mg) = labeled quantity of hydrochlorothiazide in each WH Tablet (mg) = concentration of USP Hydrochlorothiazide RS in CS the Standard solution (mg/mL) = concentration of bisoprolol fumarate in the CU Sample solution (mg/mL) F = relative response factor, see Impurity Table 1. Acceptance criteria Individual impurities: See Impurity Table 1.
Impurity Table Relative Retention Time 0.69 1.2 1.0 Relative Response Factor 1.2 1.4 Acceptance Criteria, NMT (%) 1.0 2.0
= concentration of USP Hydrochlorothiazide RS in the Standard solution (mg/mL) = nominal concentration of hydrochlorothiazide in CU the Sample solution B (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 75 rpm Times: 20 min for bisoprolol fumarate; 30 min for hydrochlorothiazide Solution A: Triethylamine and water (1:499), adjust with phosphoric acid to a pH of 3.0 Mobile phase: Acetonitrile and Solution A (1:4) Standard stock solution A: 0.5 mg/mL of USP Bisoprolol Fumarate RS in Medium Standard stock solution B: Transfer 30 mg of USP Hydrochrolothiazide RS to a 50-mL volumetric flask, dissolve with 5 mL of methanol, and dilute with Medium to volume (0.6 mg/mL). Standard solution: Dilute Standard stock solution A and Standard stock solution B in Medium to obtain a solution having known concentrations of bisoprolol fumarate and hydrochlorothiazide corresponding to those of the solution under test. Sample solution: Sample per Dissolution 711 Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 227 nm for bisoprolol and 272 nm for hydrochlorothiazide Column: 3.9-mm 15-cm; packing L11 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantities, in mg, of (C18H31NO4)2 C4H4O4 and C7H8ClN3O4S2 dissolved. Tolerances: NLT 80% (Q) of the labeled amount of (C18H31NO4)2 C4H4O4 and NLT 80% (Q) of the labeled amount of C7H8ClN3O4S2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements with respect to bisoprolol fumarate and to hydrochlorothiazide IMPURITIES Organic Impurities PROCEDURE Diluent, Solution A, Solution B, Mobile phase, System suitability solution, and Sample stock solution: Proceed as directed in the Assay. Standard solution: 2 g/mL of USP Hydrochlorothiazide RS in Diluent Sample solution: 100 g/mL of bisoprolol fumarate from Sample stock solution in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 260 nm Column: 8-mm 10-cm; packing L11 Flow rate: 3 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.5 between chlorothiazide and hydrochlorothiazide in the System suitability solution
ru
Impurity Name Compound 1 Compound 2 Hydrochlorothiazide
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Bisoprolol Fumarate RS USP Chlorothiazide RS USP Hydrochlorothiazide RS
Bleomycin Sulfate
(Comment on this Monograph)id=m9835=Bleomycin Sulfate=BMonos.pdf) Bleomycin sulfate (salt) [9041-93-4]. DEFINITION Bleomycin Sulfate is the sulfate salt of bleomycin, a mixture of basic cytotoxic glycopeptides produced by the growth of Streptomyces verticillus, or produced by other means. It has a potency of NLT 1.5 Bleomycin Units/mg and NMT 2.0 Bleomycin Units/mg. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sulfate 191 ASSAY PROCEDURE Sample solution: Dissolve a suitable quantity of Bleomycin Sulfate, in Buffer No. 16, and dilute with Buffer No. 16 to obtain a solution having a convenient concentration. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81, using an measured volume of Sample solution diluted with Buffer No. 16 to yield a Sample Dilution having
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a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 1.52.0 Bleomycin Units/mg OTHER COMPONENTS CONTENT OF BLEOMYCINS Mobile phase: 960 mg of sodium 1-pentanesulfonate in 1000 mL of deaerated 0.08 N acetic acid, adjust with ammonium hydroxide to a pH of 4.3, filter, and degas. [NOTE1.86 g of edetate disodium may be included if needed to obtain satisfactory chromatography.] Use a linear gradient of 10%40% methanol mixed with this solution, with a gradient mixing time of 60 min, and allow chromatography to proceed with the final gradient mixture for a further 20 min or until demethylbleomycin A2 has been eluted. Sample solution: 2.5 Bleomycin Units/mL from Bleomycin Sulfate in deaerated water [NOTEStore this solution in a refrigerator until just prior to use.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 250-mm stainless steel; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L Analysis Sample: Sample solution Analysis: The elution order is bleomycinic acid, bleomycin A2 (major peak), bleomycin A5, bleomycin B2 (major peak), bleomycin B4, and demethylbleomycin A2. Calculate the percentage contents of bleomycin A2, bleomycin B2, and bleomycin B4 taken: Result = (ru/rT) 100 = peak response corresponding to the particular bleomycin rT = total of the responses of all peaks Acceptance criteria: The content of bleomycin A2 is 55%70%; the content of bleomycin B2 is 25%32%; the content of bleomycin B4 is NMT 1%; and the combined percentage of bleomycin A2 and bleomycin B2 is NLT 90%. IMPURITIES Inorganic Impurities COPPER: NMT 0.1% Solution A: Nitric acid and water (1:99) Standard stock solution: 1.000 g of copper to a 1000-mL volumetric flask, dissolve in 20 mL of nitric acid, and dilute with Solution A to volume. Store in a polyethylene bottle. This solution contains 1000 g/mL of copper. Standard solutions: Transfer 5.0 mL of Standard stock solution to a 100-mL volumetric flask, and dilute with Solution A to volume. Transfer 3.0, 9.0, and 15.0 mL, respectively, of Standard stock solution to separate 100-mL volumetric flasks, dilute the contents of each flask with Solution A to volume, and mix. These Standard solutions contain, respectively, 1.5, 4.5, and 7.5 g/mL of copper. Sample solution: 7.5 mg/mL of Bleomycin Sulfate in Solution A Spectrometric conditions Mode: Atomic absorption spectrophotometry (see Spectrophotometry and Light-Scattering 851), equipped with a copper hollow-cathode lamp and an airacetylene flame Analytical wavelength: Copper emission line at 324.8 nm ru
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Blank: Solution A Analysis Samples: Standard solutions, Sample solution, and Blank Analysis: Plot the absorbances of the Standard solutions versus concentration, in g/mL, of copper, and draw the straight line best fitting the three plotted points. Calculate the percentage of copper in the portion of Bleomycin Sulfate: Result = C/W 100 C W = concentration of copper from the graph obtained, in the Sample solution (g/mL) = weight of Bleomycin Sulfate taken to prepare the Sample solution (mg)
SPECIFIC TESTS PH 791: 4.56.0, in a solution containing 10 Bleomycin Units/mL LOSS ON DRYING 731: Dry it in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 6.0% of its weight. OTHER REQUIREMENTS Where the label states that Bleomycin Sulfate is sterile, it meets the requirements for the following tests: Bacterial Endotoxins Test 85: NMT 10.0 USP Endotoxin Units/Bleomycin Unit Sterility Tests 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. Where the label states that Bleomycin Sulfate must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements for: Bacterial Endotoxins Test 85: NMT 10.0 USP Endotoxin Units/Bleomycin Unit ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Bleomycin Sulfate RS USP Endotoxin RS
Bleomycin for Injection

(Comment on this Monograph)id=m9840=Bleomycin for Injection=B-Monos.pdf) DEFINITION Bleomycin for Injection contains an amount of Bleomycin Sulfate equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of bleomycin. ASSAY PROCEDURE Sample solution: Constitute Bleomycin for Injection as directed in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and quantitatively dilute with Buffer No. 16 to obtain a solution having a convenient concentration. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81, using the Sample solution diluted quantitatively and stepwise with Buffer No. 16 to yield a dilution having a
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Bleomycin / Official Monographs
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Adjust with ammonium hydroxide to a pH of 4.3. [NOTE1.86 g of edetate disodium may be included if needed to obtain satisfactory chromatography.] Use a linear gradient of 10% to 40% methanol mixed with this solution, with a gradient mixing time of 60 min, and allow chromatography to proceed with the final gradient mixture for a further 20 min or until demethylbleomycin A2 has been eluted. Sample solution: 2.5 Bleomycin Units/mL from Bleomycin for Injection in water [NOTEStore this solution in a refrigerator until just prior to use.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 250-mm stainless steel; packing L1 Flow rate: 1.2 mL/min Injection size: 10 L Analysis Sample: Sample solution The elution order is bleomycinic acid, bleomycin A2 (major peak), bleomycin A5, bleomycin B2 (major peak), bleomycin B4, and demethylbleomycin A2. Calculate the percentage contents of bleomycin A2, bleomycin B2, and bleomycin B4: Result = (ru/rT) 100 = peak response corresponding to the particular bleomycin = sum of the responses of all peaks rT Acceptance criteria The content of bleomycin A2 is 55%70%; the content of bleomycin B2 is 25%32%. The combined percentage of bleomycin A2 and bleomycin B2 is NLT 90%. The content of bleomycin B4 is NMT 1%. OTHER REQUIREMENTS: Meets the requirements for Identification tests, under Bleomycin Sulfate and for Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Bleomycin Sulfate RS USP Endotoxin RS ru
concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
IMPURITIES Inorganic Impurities COPPER Solution A: Nitric acid and water (1:100) Standard stock solution: Transfer 1.000 g of copper to a 1000-mL volumetric flask, dissolve in 20 mL of nitric acid, dilute with Solution A to volume, and mix. Store in a polyethylene bottle. This solution contains 1000 g of copper per mL. Standard solutions: 1.5 g/mL, 4.5 g/mL, and 7.5 g/mL from Standard stock solution in Solution A Sample solution: 7.5 mg/mL of Bleomycin for Injection in Solution A Spectrometric conditions Mode: Atomic absorption spectrophotometry equipped with a copper hollow-cathode lamp and an airacetylene flame (see Spectrophotometry and Light-Scattering 851) Analytical wavelength: The copper emission line at 324.8 nm Analysis Samples: Blank (Solution A), Sample solution, and Standard solution Plot the absorbances of the Standard solutions versus concentration of copper, in g/mL, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration (C) of copper, in g/mL, in the Sample solution. Calculate the percentage of copper in the portion of Bleomycin Sulfate taken: Result = (C/CU) F 100 = concentration of copper measured in the Sample solution (g/mL) = concentration of Bleomycin Sulfate for injection CU in the Sample solution (mg/mL) F = unit conversion factor (0.001 [mg/g]) Acceptance criteria: NMT 0.1% SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: NMT 10.0 USP Endotoxin Units/Bleomycin Unit STERILITY TESTS 71: Meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration, the entire contents of each container being used. WATER DETERMINATION, Method Ic 921 Sample solution: Use a dry syringe to inject 4 mL of anhydrous methanol through the stoppers of each of two tared containers, respectively, and shake to dissolve. Using the same syringe, aspirate the contents of the two containers, transfer to the titration vessel, and titrate. Perform a blank determination on 8 mL of the anhydrous methanol. Determine the weights of the empty containers, and calculate the percentage of water. Acceptance criteria: NMT 6.0% PH 791: 4.56.0 in a solution containing 10 Bleomycin Units/mL CONTENT OF BLEOMYCINS Mobile phase: 960 mg of sodium 1-pentanesulfonate in 1000 mL of 0.08 N acetic acid C
Anti-A Blood Grouping Serum

(Comment on this Monograph)id=m9880=Anti-A Blood Grouping Serum=B-Monos.pdf) DEFINITION Anti-A Blood Grouping Serum conforms to the regulations of the federal Food and Drug Administration concerning biologics (660.20 to 660.29) (see Biologics 1041). It is a sterile, liquid or dried preparation containing the particular blood group antibodies derived from high-titered blood plasma or serum of human subjects, with or without stimulation by the injection of Blood Group Specific Substance A (or AB). It agglutinates human red cells containing A-antigens, i.e., blood groups A and AB (including subgroups A1, A2, A1B, and A2B but not necessarily weaker subgroups). It contains a suitable antimicrobial preservative. It meets the requirements to the test for potency, in parallel with, and not less than equivalent to, the U.S. Reference Blood Grouping Serum Anti-A, in
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agglutinating red blood cells from Group A1 and Group A2B donors. It meets the requirements of the tests for specificity with Group A1, A2B, B, and O cells and confirms the absence of contaminating antibodies reactive with Mg, Wra antigens as well as other antigens having an incidence of 1% or greater in the general population (see Biologics 1041, Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e). It meets the requirements of the tests for avidity with Group A1 and A2B cells. All fresh or frozen red blood cell suspensions used for these tests are prepared under specified conditions and meet specified criteria. Anti-A Blood Grouping Serum may be artificially colored blue. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve at a temperature between 2 and 8. EXPIRATION DATE: The expiration date for liquid Serum is not later than 1 year, and for dried Serum not later than 5 years after date of issue from manufacturers cold storage (5, 1 year; or 0, 2 years), provided that the expiration date for dried Serum is not later than 1 year after constitution. LABELING: Label it to state that the source material was not reactive for hepatitis B surface antigen, but that no known test method offers assurance that products derived from human blood will not transmit hepatitis. Label it also to state that it is for in vitro diagnostic use. [NOTEThe labeling is in black lettering imprinted on paper that is white or is colored completely or in part to match the specified blue color standard.]
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human blood will not transmit hepatitis. Label it also to state that it is for in vitro diagnostic use. [NOTEThe labeling is in black lettering imprinted on paper that is white or is colored completely or in part to match the specified yellow color standard.]
Blood Grouping Serums

(Comment on this Monograph)id=m9900=Blood Grouping Serums=B-Monos.pdf) DEFINITION [NOTEThis monograph deals with those Blood Grouping Serums for which there are no individual monographs and which are not routinely used or required for the testing of blood or blood products for transfusion.] Blood Grouping Serums conform to the regulations of the federal Food and Drug Administration concerning biologics (see Sections 660.20 to 660.29; see also Biologics 1041). Each is a sterile, liquid or dried preparation containing one or more of the particular blood group antibodies derived from hightitered blood plasma or serum of human subjects, with or without stimulation by the injection of red cells or other substances, or of animals after stimulation by substances that cause such antibody production. It causes, either directly or indirectly, by the antiglobulin test, the visible agglutination of human red cells containing the particular antigen(s) for which it is specific. It contains a suitable antimicrobial preservative. It meets the requirements of the test for potency, (1) in the case of tube test reagents, when tested by the specified method, of agglutinating red blood cells containing the specified antigens with the specified degree of reactivity, as defined, as follows: not less than a 1+ reaction (i.e., agglutinated cells dislodged into finely granular, but definite, small clumps) with a 1:8 dilution of Serum for Anti-K, Anti-k, Anti-Jka, Anti-Fya, Anti-Cw; NLT a 1+ reaction with a 1:4 dilution of Serum for Anti-S, Antis, Anti-P1, Anti-M, Anti-I, Anti-e (saline), Anti-c (saline) and Anti-A1; and not less than a 2+ reaction (i.e., agglutinated cells dislodged into many small clumps of equal size) with undiluted Serum for Anti-U, Anti-Kpa, Anti-Kpb, Anti-Jsa, AntiFyb, Anti-N, Anti-Lea, Anti-Leb, Anti-Dia, Anti-Mg, Anti-Jkb, and Anti-Xga; and (2) in the case of reagents recommended for slide test methods, of agglutinating red blood cells, with the specified degree of reactivity, as defined, with both undiluted Serum and with a 1:2 dilution of Serum, when tested by the manufacturers recommended method(s) using cells heterozygous for the corresponding antigen(s). It meets the requirements of the tests for specificity by the most sensitive method recommended by the manufacturer, in which not less than 4 positive and 4 negative phenotypes are included, and confirms the absence of contaminating antibodies reactive with Mg, Wra antigens as well as other antigens having an incidence of 1% or greater in the general population (see Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e). It meets the requirements of the tests for avidity with the manufacturers recommended method, red blood cells heterozygous for the corresponding antigen(s) being used. All fresh or frozen red blood cell suspensions used for these tests are prepared under specified conditions and meet specified criteria. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve at a temperature between 2 and 8. EXPIRATION DATE: The expiration date for liquid Serum is not later than 1 year and for dried Serum not later than 5 years after date of issue from manufacturers cold storage (5, 1 year; or 0, 2 years), provided that the expiration date for dried Serum is not later than 1 year after constitution.
Anti-B Blood Grouping Serum

(Comment on this Monograph)id=m9890=Anti-B Blood Grouping Serum=B-Monos.pdf) DEFINITION Anti-B Blood Grouping Serum conforms to the regulations of the federal Food and Drug Administration concerning biologics (660.20 to 660.29) (see Biologics 1041). It is a sterile, liquid or dried preparation containing the particular blood group antibodies derived from high-titered blood plasma or serum of human subjects, with or without stimulation by the injection of Blood Group Specific Substance B (or AB). It agglutinates human red cells containing B-antigens, i.e., blood groups B and AB (including subgroups A1B and A2B). It contains a suitable antimicrobial preservative. It meets the requirements of the test for potency, in parallel with, and not less than equivalent to, the U.S. Reference Blood Grouping Serum AntiB, in agglutinating red blood cells from Group B donors. It meets the requirements of the tests for specificity with Group A1, B, and O cells and confirms the absence of contaminating antibodies reactive with Mg, Wra antigens as well as other antigens having an incidence of 1% or greater in the general population (see under Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e). It meets the requirements of the test for avidity with Group B cells. All fresh or frozen red blood cell suspensions used for these tests are prepared under specified conditions and meet specified criteria. Anti-B Blood Grouping Serum may be artificially colored yellow. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve at a temperature between 2 and 8. EXPIRATION DATE: The expiration date for liquid Serum is not later than 1 year and for dried Serum not later than 5 years after date of issue from manufacturers cold storage (5, 1 year; or 0, 2 years), provided that the expiration date for dried Serum is not later than 1 year after constitution. LABELING: Label it to state that the source material was not reactive for hepatitis B surface antigen, but that no known test method offers assurance that products derived from
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Table 2 Serum Anti-D Anti-C Anti-E Anti-CD Anti-DE Anti-CDE Anti-c Anti-e Phenotype of Cells cDe Ccde cdEe cDe, Ccde cDe, cdEe cdEe, cDe, Ccde CcDEe cdEe
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LABELING: Label each to state the source of the product if other than human and, if of human origin, to state that the source material was not reactive for hepatitis B surface antigen, but that no known test method offers assurance that products derived from human blood will not transmit hepatitis. Label each also to state that it is for in vitro diagnostic use.
Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e

(Comment on this Monograph)id=m9910=Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e=B-Monos.pdf) Anti-Rh Blood Grouping Serums DEFINITION Blood Grouping Serums Anti-D, Anti-C, Anti-E, Anti-c, Anti-e (Anti-Rh Group) conform to the regulations of the federal Food and Drug Administration concerning biologics (660.20 to 660.29) (see Biologics 1041). They are sterile, liquid, or dried preparations derived from the blood plasma or serum of human subjects who have developed specific Rh antibodies. They are free from agglutinins for the A or B antigens and from alloantibodies other than those for which claims are made in the labeling. They contain a suitable antimicrobial preservative. Liquid serums are not artificially colored. Two varieties of Anti-Rh Blood Grouping Serums are recognized, i.e., (1) saline agglutinating complete antiserums, which specifically agglutinate human red blood cells suspended in saline TS, and (2) blocking or incomplete antiserums, which contain protein or other macromolecular substances, usually require the cells to be suspended in serum or plasma, and generally are for slide or rapid tube tests. The most commonly used of these blood grouping serums are listed in Table 1, each reacting with the antigen(s) designated by the corresponding letter(s) with the alternative nomenclature indicated parenthetically.
Table 1 Serum Anti-D (Anti-Rho) Anti-C (Anti-rh) Anti-E (Anti-rh) Anti-CD (Anti-Rho) Anti-DE (Anti-Rho) Anti-CDE (Anti-Rho) Anti-c (Anti-hr) Anti-e (Anti-hr) Antigen(s) Reacting D (Rho) C (rh) E (rh) D (Rho) and C (rh) D (Rho) and E (rh) D (Rho), C (rh), and E (rh) c (hr) e (hr)
Each serum meets the requirements of the tests for specificity by the most sensitive method recommended by the manufacturer, in which NLT 4 positive and 4 negative phenotypes are included (see Table 3), and confirms the absence of contaminating antibodies reactive with Mg, Wra antigens as well as other antigens having an incidence of 1% or greater in the general population, except where some of these confirmatory tests cannot be done by the manufacturer, in which event such omissions are noted. Antigens having such population incidence in the United States [other than low-incidence antigens, serum proteins (e.g., Gm, Km), leukocyte factors, drugs, chemicals, or polyagglutinable cells, which are not necessarily excluded] are the following: A, B, H, Lea, Leb, Lec, Led, I, K, k, Kpa, Kpb, Jsb, P1, D, C, E, c, e, Cw, M, N, S, s, U, Lua, Lub, Jka, Jkb, Fya, Fyb, Xga, Doa, Dob, Yta, Ytb, Lan, Coa, Cob, Mg, Wra, and Sda.
Table 3 Serum Anti-D Cells CcDe, cDe, Ccde, cdEe, A1 cde, B cde, O cde, and where recommended for use by indirect antiglobulin technique, cde Bg(a+) cells from 3 different donors cDe, Ccde, cdEe, C + rhi neg. cells, A1 cde, B cde, O cde cDe, Ccde, cdEe, A1 cde, B cde, O cde cDe, Ccde, cdEe, A1 cde, B cde, O cde, and where recommended for detection of the G antigen, rGr cDe, Ccde, cdEe, A1 cde, B cde, O cde cDe, Ccde, cdEe, A1 cde, B cde, O cde, and where recommended for detection of the G antigen, rGr Ccde, A1 CDe, B CDe, O CDe, and CDEe or CDE or CdE cdEe, A1 cDE, B cDE, O cDE, and CcDE or CDE or CdE
Anti-C Anti-E Anti-CD
Anti-DE Anti-CDE
Anti-c Anti-e
Each Serum meets the requirements of the test for potency in the case of serums for saline tube test in parallel with, and NLT equivalent to, the U.S. Reference Blood Grouping Serum for Anti-D, Anti-C, or Anti-E, whichever is applicable, or, in the case of Anti-c and Anti-e for saline tube test which have no reference preparations, the test for minimum agglutination reactivity at a specified dilution; and in the case of serums for slide or rapid tube test in parallel with, and NLT than equivalent to, the U.S. Reference Blood Grouping Serum for Anti-D, Anti-C, Anti-E, Anti-c, or Anti-e, whichever is applicable, in agglutinating as a minimum red blood cells from the donors indicated in Table 2 (which may be from Group A, B, AB, or O). Each serum for slide or rapid tube test meets the requirements of the tests for avidity with the cells as indicated under tests for potency above.
All fresh or frozen red blood cell suspensions used for these tests are prepared under specified conditions and meet specified criteria. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve at a temperature between 2 and 8. EXPIRATION DATE: The expiration date for liquid Serums is not later than 1 year and for dried Serums not later than 5 years after date of issue from manufacturers cold storage (5, 1 year; or 0, 2 years), provided that the expiration date for dried Serums is not later than 1 year after constitution. LABELING: Label each to state that the source material was not reactive for hepatitis B surface antigen, but that no
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known test method offers assurance that products derived from human blood will not transmit hepatitis. Label each to state that it is for in vitro diagnostic use.

Analysis Samples: Control solutions and Sample solution On a suitable U-bottomed microtiter plate, place 1 drop of 0.9% saline in each of three different wells in a row (Blank Row). Place 1 drop from one of the lots of Anti-A reagent in each of three different wells in a second row. Place 1 drop from the second lot of Anti-A reagent in each of three different wells in a third row. Repeat the same with two different lots of Anti-B reagent and one lot of Anti-AB reagent in separate rows. To each row, add 1 drop of Control solution A1, Control solution B, and the Sample solution in the first, second, and the third well, respectively, of each row. Mix the contents of the wells by gently tapping the sides of the plate. Centrifuge the plate at the appropriate conditions established for the centrifuge. Resuspend the cell buttons by manually tapping the plate, or with the aid of a suitable mechanical shaker. Read the optical densities at different wells using a suitable automated photometric microtiter plate reader. Compare the optical density of each well in the Blank Row with the optical density of the wells to which the corresponding Control solution A1, Control solution B, or Sample solution were added. [NOTEA high optical density comparable to those obtained for the wells in the Blank Row indicates negative results (no hemagglutination), which can be corroborated by visual observation of smooth suspensions. A low optical density indicates positive results (hemagglutination), which can be corroborated by visual observation of formation of clumps.] The blood group of Red Blood Cells is A, B, AB, or O, accordingly, as the Sample solution is hemagglutinated by Anti-A reagent, Anti-B reagent, both reagents, or neither, respectively. The blood group of the Sample solution conforms to the blood group indicated on the label. The test is not valid if Control solutions for Blood Group A and Blood Group B red blood cells are not agglutinated by Anti-A reagent and Anti-B reagent, respectively, or if both Control solutions are not agglutinated by Anti-AB reagent. The test is also not valid if the Sample solution is not agglutinated by Anti-AB reagent but is agglutinated by either Anti-A reagent or Anti-B reagent, or is agglutinated by Anti-AB reagent but not by either Anti-A reagent or Anti-B reagent. B. RH TYPE Test 1 Anti-D (Rho) reagent: Use anti-D (Rho) blood grouping reagent approved for use in microtiter plate tests. Dilute, if necessary, following the manufacturers instructions. Sample solution: Prepare as directed for Identification test A. Analysis: On a suitable U-bottom microtiter plate, place 1 drop each of 0.9% saline and Anti-D (Rho) reagent in two separate wells. Label them as the B well (Blank) and the T well, respectively. Add 1 drop of Sample solution to each well, and mix by gently tapping the side of the plate. Centrifuge the plate at appropriate conditions established for the centrifuge. Resuspend the cell buttons by manually tapping the plate, or with the aid of a suitable mechanical shaker. Read the optical densities of the wells using a suitable automated photometric microtiter plate reader, and determine if the Red Blood Cells in the T well are agglutinated as described under Identification test A. If the T well indicates negative results (no hemagglutination), incubate the plate at 37 for 15 min, centrifuge, resuspend the cells, and read the optical densities of the wells as above. Agglutination of cells after immediate-spin or after 37 incubation indicates Rh positive typing of Red Blood Cells. The test is valid if cells in the B well are not agglutinated. If the cells are not agglutinated in the T well, proceed as directed in Test 2.
Whole Blood
(Comment on this Monograph)id=m9950=Whole Blood=BMonos.pdf) ACD Whole Blood CPD Whole Blood CPDA-1 Whole Blood Heparin Whole Blood DEFINITION Whole Blood conforms to the regulations of the federal Food and Drug Administration concerning biologics (21 CFR 640.1 to 640.6) (see Biologics 1041). It is blood that has been collected from suitable human donors under rigid aseptic precautions, for transfusion to human recipients or for further processing into one or more of its components for transfusion. It contains a citrate-based anticoagulant. Whole Blood must be tested for syphilis, hepatitis B virus, Human T-Cell Lymphotropic Virus (HTLV) type I and type II, and tested for blood group and Rh factors and unexpected antibodies to red cell antigens using FDA-licensed commercially available test kits, the results of which must be below the approved upper limit of detection specified in the respective test kits. In addition, Whole Blood must also be tested for hepatitis C and HIV using FDA-approved nucleic acid assays, the results of which must be below the approved upper limit of detection for the method. A unit (dose) of Whole Blood contains a minimum of 50 g (based on a minimum donor hematocrit of 38%) of hemoglobin in a total volume of 450 mL 10% or 500 mL 10% as indicated on the label. Whole Blood may be further processed. When filtered for removal of leukocytes, the quantity of residual leukocytes in the unit of Whole Blood must be less than 5 106. IDENTIFICATION A. ABO BLOOD GROUP Anti-A reagent: Use approved commercially available monoclonal or polyclonal anti-A blood grouping reagent, two different lots from the same or different manufacturers. Use in accordance with manufacturers instructions. Anti-B reagent: Use approved commercially available monoclonal or polyclonal anti-B blood grouping reagent, two different lots from the same or different manufacturers. Use in accordance with manufacturers instructions. Anti-AB reagent: Use approved commercially available antiAB blood grouping reagent. Use in accordance with manufacturers instructions. Control solutions: On the day of use, dilute Blood Group A1 (Control solution A1) and Blood Group B (Control solution B) red blood cells, obtained from an approved commercial source or prepared by the testing laboratory, with 0.9% saline to suspensions of approximately the same concentration between 2% to 5% (v/v). [NOTEIf the Blood Group A1 and Blood Group B red blood cells are prepared in the testing laboratory from whole blood of a known blood group, it must be prepared on the day of use following the procedure for the Sample solution under Whole Blood.] Sample solution: Centrifuge at 4 a suitable volume of Whole Blood at 5000 g for 5 min. Remove the plasma from the top, taking care not to disturb the pellet of red blood cells at the bottom. Add 0.9% saline to obtain a final volume that is equal to the volume of Whole Blood. Resuspend the pellet, and centrifuge as above. Repeat the procedure once more. Dilute the red blood cells with 0.9% saline to a suspension to obtain a concentration of red blood cells that is the same as those of the Control solutions.
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Sample solution to each of T1 and T2, rinsing the pipet tip three to four times with Drabkins solution, and mix. Allow to stand for at least 15 min at room temperature. [NOTE Red Blood cells with appreciable carboxyhemoglobin content, such as those obtained from smokers, may require longer reaction time. If the donor characteristics are not known, the incubation time should be optimized prior to testing]. Read the absorbances of the solutions against the solution in tube B1 at 540 nm. The absorbance of the solution in tube B2 is recorded at the end. The test is not valid if the absorbance of the solution in tube B2 is not within 0.005. Calculations: Calculate the concentrations, in mg/mL, of hemoglobin in Standard solutions A, B, and C. Plot a calibration curve of absorbance values against the hemoglobin concentration by drawing a best-fit straight line using the least-square linear regression analysis. From the absorbance value of the Sample solution, obtain the concentration, in mg/mL, of hemoglobin in the Sample solution. Calculate the total hemoglobin content in the Red Blood Cells unit, in g, by the formula: Conc. of hemoglobin (mg/mL) the volume of the Red Blood Cells unit (mL)/103 Leukocyte count (for units labeled as Whole Blood, Leukocytes Reduced) Sample solution: Pipet 40 L of a suitable red cell-lysing agent into a clean test tube, add 100 L of Whole Blood diluted with 0.9% saline, if necessary, such that the hematocrit of Red Blood Cells is not greater than 60%. Analysis: Mix by pipetting up and down several times. Add 360 L of 0.01% (w/v) crystal violet in 15% (v/v) acetic acid into the mixture, and mix thoroughly. Fit a hemocytometer with a 50-L counting volume and a bright background, with a cover slip, and load the counting chamber with the mixture until the counting area is completely covered, but not overfull. Cover the counting chamber with a suitable moist lid to prevent evaporation, and allow to settle undisturbed for 10 to 15 min. Remove the lid, place the chamber on the stage of a light microscope fitted with 10 ocular lens and 20 objective. Count the leukocytes in the entire 50-L counting volume. Calculate the leukocyte count in Red Blood Cells, expressed in leukocytes/L, by dividing the observed leukocyte count by 10. Calculate the total number of leukocytes in the Red Blood Cells unit by using the following formula: Total leukocytes = leukocytes/L 103 volume of Red Blood Cells unit (mL) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Collect into an approved container (see Transfusion and Infusion Assemblies and Similar Medical Devices 161) containing a sterile, pyrogen-free approved anticoagulant (see USP monographs Anticoagulant Citrate Dextrose Solution, Anticoagulant Citrate Phosphate Dextrose Solution, or Anticoagulant Citrate Phosphate Dextrose Adenine Solution). Store Whole Blood in the original container, or transfer to an equivalent one using a technique that does not compromise sterility. Whole Blood is stored at a temperature between 1 and 6 unless platelets are to be prepared, in which case the blood is stored for no longer than 8 h following collection at room temperature. (See General Notices and Requirements.) EXPIRATION DATE: Whole Blood collected in Anticoagulant Citrate Dextrose Solution, Anticoagulant Citrate Phosphate Dextrose Solution, or in Anticoagulant Citrate Phosphate DextroseDextrose Solution may be stored for up to 21 days at 16 after the blood has been drawn. Whole Blood collected in Anticoagulant Citrate Phosphate Dextrose Adenine Solution may be stored for up to 35 days at 16.
Test 2 Anti-D reagent: Use anti-D blood grouping reagent approved for use for a weak D blood group test. Antihuman globulin reagent: Use polyspecific or anti-IgG antihuman globulin reagent. Dilute, if necessary, following the manufacturers instructions. Control solution: Use IgG-coated red cells approved for use as a control for Rh typing. Dilute with 0.9% saline to obtain a 2% solution. Sample solution: Prepare as directed for Test 1. Analysis: Place 1 drop of 0.9% saline into a suitable test tube and 1 drop of Anti-D reagent in another, and mark them as the Blank and Anti-D, respectively. To each tube, add 1 drop of the Sample solution, mix, and incubate at 37 for 15 to 30 min. Centrifuge the tubes at 1000 g for 15 to 30 s. Gently resuspend the cell buttons, and examine them for hemagglutination (formation of clump) by visual examination. The Rh typing of Red Blood Cells is positive or negative according to whether the cells in Anti-D tube are agglutinated or not. If the cells are not agglutinated, add 1 mL of 0.9% saline to the Anti-D tube, and resuspend the cells. Centrifuge the tube at 1000 g for 1 min, and remove the saline completely. Repeat the step two to three times more to wash the Red Blood Cells. Add 1 drop of 0.9% saline and 1 to 2 drops of Antihuman globulin reagent to the Anti-D tube. Mix gently, and centrifuge the tube at 1000 g for 15 to 30 s. Gently resuspend the cell button, add 1 drop of Control solution, mix gently, centrifuge as above, and examine for agglutination. The Rh Type of the Sample solution conforms to the Rh Type indicated on the label. The test is not valid if the cells in the Anti-D tube are not agglutinated after addition of the Control solution. Also, for the test to be valid, the cells in the Blank tube must not be agglutinated. SPECIFIC TESTS VISUAL INSPECTION: Inspect visually during storage and immediately prior to use. If the color or physical appearance is abnormal or there is any indication or suspicion of microbial contamination, the unit is unsuitable for transfusion. HEMOGLOBIN CONTENT Drabkins solution: Dissolve Drabkins reagent in a suitable volume of water, and add a suitable volume of a 30% (w/v) polyoxyethylene (23) lauryl ether solution such that the final concentrations of potassium cyanide, potassium ferrocyanide, and polyoxyethylene (23) lauryl ether in the solution are approximately 0.75 mM, 0.6 mM, and 0.015%, respectively. Store the solution in the dark between 1826. [CautionDrabkins reagent and Drabkins solution contain cyanide and are HIGHLY TOXIC. Do not inhale or swallow or allow contact with skin or eyes. Wear suitable protective clothing, gloves, and eye and face protection. Do not mix with acids. Contact with acids liberates a very toxic gas. If ingested, perform gastric lavage, and immediately call a physician.] Blank solution: Water Standard solution A: 300 mg of USP Hemoglobin RS in a 2-mL volumetric tube, add 1 mL water, dissolve in and dilute with water to volume. Standard solution B: Mix 50 L of Standard solution A with 25 L of water. Standard solution C: Mix 50 L of Standard solution A with 100 L of water. Sample solution: 20 L of Whole Blood (without dilution) Analysis: Label suitable tubes as B1, B2, SA1, SA2, SB1, SB2, SC1, SC2, T1, and T2. Add 5.0 mL of Drabkins solution to each tube. Add 20 L of water to each of B1 and B2, 20 L of Standard solution A to each of SA1 and SA2, 20 L of Standard solution B to each of SB1 and SB2, 20 L of Standard solution C to each of SC1 and SC2, and 20 L of
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If the hermetic seal of the container is broken during collection, solution, or further processing, the expiration date is not later than 24 h after the seal is broken (when blood is stored at 16), but not to exceed the original expiration date of the unit. LABELING: Label the container to indicate the volume of the Whole Blood collected from the donor, the collection date, the donation number or other coding means to uniquely identify the unit and to provide traceability to the donor, the storage temperature, and the expiration date (see below). Label to indicate the type of anticoagulant or other preservative solution used to collect it. Label also to identify donor status (i.e., volunteer or paid). Label also with the following statements: See Circular of Information for indications, contraindications, cautions, and methods of infusion.; Properly identify recipient; and [CAUTIONRx only.] Label to indicate the ABO group and Rh type, as indicated in Table 1. [NOTEEach Whole Blood product must have a determination made as to its ABO group and Rh type and specificity of unexpected red cell antibodies, if any.]
Table 1 ABO Group A A B B AB AB O O Rh Type Positive Negative Positive Negative Positive Negative Positive Negative

approved and licensed commercially available test kits, the results of which must be below the limits of detection specified in the respective test kits by the manufacturers. In addition, the source blood must also be tested for hepatitis C and HIV using FDA-approved nucleic acid assays, the results of which must be below the approved limits of detection for the method. A unit (dose) of Red Blood Cells contains a minimum of 50 g of hemoglobin in a total volume of about 180325 mL. A unit (dose) of Red Blood Cells, Leukocytes Reduced contains a minimum of 42.5 g of hemoglobin in a total volume of about 150275 mL, and has a residual leukocyte count of less than 5 106. A unit (dose) of Red Blood Cells, Deglycerolized contains a minimum of 40 g of hemoglobin in a total volume of about 180325 mL. A unit (dose) of Red Blood Cells, Leukocytes Reduced and Deglycerolized contains a minimum of 34 g of hemoglobin in a total volume of approximately 180325 mL and has a residual leukocyte count of less than 5 106. A unit (dose) of Red Blood Cells, Pheresis contains a mean hemoglobin content of 60 g of hemoglobin. A unit (dose) of Red Blood Cells, Pheresis, Leucocytes Reduced contains a mean hemoglobin content of 51 g (or 153 mL packed red cell volume) and has a residual leukocyte count of less than 5 106. IDENTIFICATION A. ABO BLOOD GROUP Anti-A reagent: Use approved commercially available monoclonal or polyclonal anti-A blood grouping reagent, two different lots from the same or different manufacturers. Use in accordance with manufacturers instructions. Anti-B reagent: Use approved commercially available monoclonal or polyclonal anti-B blood grouping reagent, two different lots from the same or different manufacturers. Use in accordance with manufacturers instructions. Anti-AB reagent: Use approved commercially available antiAB blood grouping reagent. Use in accordance with manufacturers instructions. Control solutions: On the day of use, dilute Blood Group A1 (Control solution A1) and Blood Group B (Control solution B) red blood cells, obtained from an approved commercial source or prepared by the testing laboratory, with 0.9% saline to suspensions of about the same concentration between 2% and 5%. [NOTEIf the Blood Group A1 and Blood Group B red blood cells are prepared in the testing laboratory from whole blood of a known blood group, it must be prepared on the day of use according to the following procedure: Centrifuge at 4 a suitable volume of Whole Blood at 5000 g for 5 min. Remove the plasma from the top, taking care not to disturb the pellet of red blood cells at the bottom. Add 0.9% saline to obtain a final volume that is about equal to the volume of Whole Blood. Resuspend the pellet, and centrifuge as above. Repeat the procedure once more. Dilute the red blood cells with 0.9% saline to a suspension to obtain a concentration of red blood cells that is about the same as those of the Control solutions.] Sample solution: On the day of use, dilute Red Blood Cells with 0.9% saline to a suspension of about the same concentration as the Control solutions. Analysis Samples: Control solutions and Sample solution On a suitable U-bottomed microtiter plate, place 1 drop of 0.9% saline in each of three different wells in a row (Blank Row). Place 1 drop from one of the lots of Anti-A reagent in each of three different wells in a second row. Place 1 drop from the second lot of Anti-A reagent in each of three different wells in a third row. Repeat the same with two different lots of Anti-B reagent and one lot of Anti-AB reagent in separate rows. To each row, add 1 drop of Control solution A1, Control solution B, and the Sample
If an ABO blood group color scheme is used, use the following labeling color: Group A (yellow), Group B (pink), Group O (blue), and Group AB (white). Label Whole Blood with the type and results of tests for adventitious agents. If it has been issued prior to determination of the test results, label also with the warning Donor Untested and to further specify Uncrossmatched Blood, when appropriate. If the unit of Whole Blood was filtered to reduce leukocytes, label as Whole Blood, Leukocytes Reduced. USP REFERENCE STANDARDS 11 USP Hemoglobin RS
Red Blood Cells

(Comment on this Monograph)id=m9940=Red Blood Cells=BMonos.pdf) DEFINITION Red Blood Cells is the portion of blood that contains hemoglobin and is derived from human whole blood, from which plasma and platelets are removed by centrifugation, sedimentation, or by apheresis. In the case of apheresis, the plasma is automatically removed and returned directly to the donor. Red Blood Cells derived from whole blood may be prepared at any time during the dating period of the whole blood from which it is derived. Red Blood Cells may be further processed by addition of red cell preservatives, irradiation to inactivate lymphocytes, filtration for removal of leukocytes, washing to remove proteins, freezing and thawing, or rejuvenation using validated and approved procedures. The source blood for Red Blood Cells must be tested for syphilis, hepatitis B virus, hepatitis C virus, Human T-cell Lymphotropic Virus (HTLV) type I and type II, human immunodeficiency virus (HIV) type 1 and type 2, and unexpected antibodies to red cell antigens using FDA-
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them as the Blank and Anti-D, respectively. To each tube, add 1 drop of the Sample solution, and incubate at 37 for 1530 min. Centrifuge the tubes at about 1000 g for 1530 s. Gently resuspend the cell buttons, and examine them for hemagglutination (formation of clump) by visual examination. The Rh typing of Red Blood Cells is positive or negative according to whether the cells in Anti-D tube are agglutinated or not. If the cells are not agglutinated, add 1 mL of 0.9% saline to the Anti-D tube, and resuspend the cells. Centrifuge the tube at about 1000g for 1 min, and remove the saline completely. Repeat the step two to three times more to wash the Red Blood Cells. Add 1 drop of 0.9% saline and 1 to 2 drops of Antihuman globulin reagent to the Anti-D tube. Mix gently, and centrifuge the tube at about 1000 g for 1530 s. Gently resuspend the cell button, add 1 drop of Control solution, mix gently, centrifuge as above, and examine for agglutination. The Rh Type of the Sample solution conforms to the Rh Type indicated on the label. The test is not valid if the cells in the Anti-D tube are not agglutinated after adding the Control solution. Also, for the test to be valid, the cells in the Blank tube must not be agglutinated. SPECIFIC TESTS VISUAL INSPECTION: Inspect visually during storage and immediately prior to use. If the color or physical appearance is abnormal or there is any indication or suspicion of microbial contamination, the unit is unsuitable for transfusion. HEMOGLOBIN CONTENT Drabkins solution: Dissolve Drabkins reagent in a suitable volume of water, and add a suitable volume of a 30% (w/v) polyoxyethylene (23) lauryl ether solution such that the final concentrations of potassium cyanide, potassium ferrocyanide, and polyoxyethylene (23) lauryl ether in the solution are approximately 0.75 mM, 0.6 mM, and 0.015%, respectively. Store the solution in the dark between 18 and 26. [CautionDrabkins reagent and Drabkins solution contain cyanide and are HIGHLY TOXIC. Do not inhale, swallow, or allow contact with skin or with eyes. Wear suitable protective clothing, gloves, and eye and face protection. Do not mix with acids. Contact with acids liberates a very toxic gas. If ingested, perform gastric lavage, and immediately call a physician.] Blank solution: Water Standard solution A: 300 mg USP Hemoglobin RS in a 2mL volumetric tube, add 1 mL water, then dissolve in and dilute with water to volume Standard solution B: Mix 50 L of Standard solution A with 25 L of water. Standard solution C: Mix 50 L of Standard solution A with 100 L of water. Sample solution: Mix 50 L of Red Blood Cells with 50 L of water. Analysis: Label suitable tubes as B1, B2, SA1, SA2, SB1, SB2, SC1, SC2, T1, and T2. Add 5.0 mL of Drabkins solution to each tube. Add 20 L of water to each of B1 and B2, 20 L of Standard solution A to each of SA1 and SA2, 20 L of Standard solution B to each of SB1 and SB2, 20 L of Standard solution C to each of SC1 and SC2, and 20 L of Sample solution to each of T1 and T2, rinsing the pipet tip three to four times with Drabkins solution. Allow to stand for at least 15 min at room temperature. [NOTERed Blood Cells with appreciable carboxyhemoglobin content, such as those obtained from smokers, may require longer reaction time. If the donor characteristics are not known, the incubation time should be optimized prior to testing.] Read the absorbances of the solutions against the solution in tube B1 at 540 nm. The absorbance of the solution in tube B2 is
solution in the first, second, and the third well, respectively, of each row. Mix the contents of the wells by gently tapping the sides of the plate. Centrifuge the plate at the appropriate conditions established for the centrifuge. Resuspend the cell buttons by manually tapping the plate, or with the aid of a suitable mechanical shaker. Read the optical densities at different wells using a suitable automated photometric microtiter plate reader. Compare the optical density of each well in the Blank Row with the optical density of the wells to which the corresponding Control solution A1, Control solution B, or Sample solution were added. [NOTEA high optical density comparable to those obtained for the wells in the Blank Row indicates negative results (no hemagglutination), which can be corroborated by visual observation of smooth suspensions. A low optical density indicates positive results (hemagglutination), which can be corroborated by visual observation of formation of clumps.] The blood group of Red Blood Cells is A, B, AB, or O, accordingly, as the Sample solution is hemagglutinated by Anti-A reagent, Anti-B reagent, both reagents, or neither, respectively. The blood group of the Sample solution conforms to the blood group indicated on the label. The test is not valid if Control solutions for Blood Group A and Blood Group B red blood cells are not agglutinated by Anti-A reagent and Anti-B reagent, respectively, or if both Control solutions are not agglutinated by Anti-AB reagent. The test is also not valid if the Sample solution is not agglutinated by Anti-AB reagent but is agglutinated by either Anti-A reagent or Anti-B reagent, or is agglutinated by Anti-AB reagent but not by either Anti-A reagent or Anti-B reagent. B. RH TYPE Test 1 Anti-D (Rho) reagent: Use anti-D (Rho) blood grouping reagent approved for use in microtiter plate tests. Dilute, if necessary, following the manufacturers instructions. Sample solution: On the day of use, dilute Red Blood Cells with 0.9% saline to obtain a 2%5% suspension. Analysis: On a suitable U-bottom microtiter plate, place 1 drop each of 0.9% saline and Anti-D (Rho) reagent in two separate wells. Label them as the B well (Blank) and the T well, respectively. Add 1 drop of Sample solution to each well, and mix by gently tapping the side of the plate. Centrifuge the plate at appropriate conditions established for the centrifuge. Resuspend the cell buttons by manually tapping the plate or with the aid of a suitable mechanical shaker. Read the optical densities of the wells using a suitable automated photometric microtiter plate reader, and determine if the Red Blood Cells in the T well is agglutinated as described under Identification test A. If the T well indicates negative results (no hemagglutination), incubate the plate at 37 for 15 min, centrifuge, resuspend the cells, and read the optical densities of the wells as above. Agglutination of cells after immediate-spin or after 37 incubation indicates Rh positive typing of Red Blood Cells. The test is valid if cells in the B well are not agglutinated. If the cells are not agglutinated in the T well, proceed as directed in Test 2. Test 2 Anti-D reagent: Use an anti-D blood grouping reagent approved for use for a weak D blood group test. Antihuman globulin reagent: Use a polyspecific or antiIgG antihuman globulin reagent. Dilute, if necessary, following the manufacturers instructions. Control solution: Use IgG-coated red cells approved for use as a control for Rh typing. Dilute with 0.9% saline to obtain a 2% solution. Sample solution: Prepare as directed for Test 1. Analysis: Place 1 drop of 0.9% saline into a suitable test tube and 1 drop of Anti-D reagent in another, and mark
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recorded at the end. The test is not valid if the absorbance of the solution in tube B2 is not within 0.005. Calculations: Calculate the concentrations, in mg/mL, of hemoglobin in Standard solutions A, B, and C. Plot a calibration curve of absorbance values against the hemoglobin concentration by drawing a best-fit straight line using the least-square linear regression analysis. From the absorbance value of the Sample solution, obtain the concentration, in mg/mL, of hemoglobin in the Sample solution. Multiply the value by 2 to obtain the concentration, in mg/mL, of hemoglobin in Red Blood Cells. Calculate the total hemoglobin content in the Red Blood Cells unit, in g: Result = Conc. of hemoglobin (mg/mL) the volume of the Red Blood Cells unit (mL)/103 LEUKOCYTE COUNT Sample solution: Pipet 40 L of a suitable red cell-lysing agent into a clean test tube, add 100 L of Red Blood Cells diluted with 0.9% saline, if necessary, such that the hematocrit of Red Blood Cells is not greater than 60%. Analysis: Mix by pipetting up and down several times. Add 360 L of 0.01% (w/v) crystal violet in 15% acetic acid into the mixture, and mix thoroughly. Fit a hemocytometer with a 50-L counting volume and a bright background, with a cover slip, and load the counting chamber with the mixture until the counting area is completely covered but not overfull. Cover the counting chamber with a suitable moist lid to prevent evaporation, and allow to settle undisturbed for 1015 min. Remove the lid, and place the chamber on the stage of a light microscope fitted with 10 ocular lens and 20 objective. Count the leukocytes in the entire 50-L counting volume. Calculate the leukocyte count in Red Blood Cells, expressed in leukocytes/L, by dividing the observed leukocyte count by 10. Calculate the total number of leukocytes in the Red Blood Cells unit: Result = leukocytes/L 103 the volume of the Red Blood Cells unit (mL) ADEQUACY OF DEGLYCEROLIZATION (for Red Blood Cells, Deglycerolized): Interrupt the last wash cycle of the deglycerolization process at a point where the wash fluid is visible in the clear tubing segment leading to the waste receptacle. Hold the tubing against a well-lighted, white background. Note the coloration of the fluid in the tubing, and compare it to a suitable hemolysis color comparator standard. The color of the fluid should be no stronger than the block indicating 3% hemolysis. [NOTEIf the level of hemolysis is more than 3%, continue the wash process, and repeat the test until the color is within acceptable limits.] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Collect into an approved container (see Transfusion and Infusion Assemblies and Similar Medical Devices 161) containing a sterile, pyrogen-free, approved anticoagulant (see Anticoagulant Citrate Dextrose Solution, Anticoagulant Citrate Phosphate Dextrose, or Anticoagulant Citrate Phosphate Dextrose Adenine Solution). An approved additive solution may be added after removal of the plasma. Store Red Blood Cells in the original container, or transfer to an equivalent container using a technique that does not compromise sterility. Liquid Red Blood Cells is stored at a temperature between 1 and 6 (see General Notices and Requirements). Frozen Red Blood Cells is stored at 65 or colder. EXPIRATION DATE: Red Blood Cells in Anticoagulant Citrate Dextrose Solution, in Anticoagulant Citrate Phosphate Dextrose Solution, or in Anticoagulant Citrate Phosphate DextroseDextrose Solution may be stored for up to 21 days at 16

after the blood has been drawn. Red Blood Cells in Anticoagulant Citrate Phosphate Dextrose Adenine Solution may be stored for up to 35 days at 16. Red Blood Cells may be stored in an approved additive solution (AS), for up to 42 day at 16. If the hermetic seal of the container is broken during collection, solution, or further processing, the expiration date is not later than 24 h after the seal is broken (when blood is stored at 16). The expiration date for frozen Red Blood Cells prepared with low glycerol content (20% glycerol) is not later than 10 years from the date of collection when stored at 120 or colder, except when Red Blood Cells is prepared for freezing with high glycerol content (40% glycerol), in which case it may be stored at 65 or colder for no later than 10 years from date of collection. If the frozen Red Blood Cells is processed for freezing or for thawing, in an open system, the expiration date for the thawed Red Blood Cells is 24 h after removal from 65 storage, provided it is then stored at the temperature of unfrozen Red Blood Cells. LABELING: Label the container to indicate the volume of the whole human blood collected from the donor, the collection date, the donation number or other coding means to uniquely identify the unit and to provide traceability to the donor, and the expiration date. Label it to indicate the type of anticoagulant used to collect whole human blood and any additive solutions added subsequent to collection. Label it also to identify donor status (i.e., volunteer or paid). Label it also with the following statements: See Circular of Information for indications, contraindications, cautions, and methods of infusion; Properly identify recipient; and Caution: Rx only. In addition, label it to indicate the product name as indicated in Table 1. [NOTEThe name is determined by the method of solution of the Red Blood Cells (derived from whole human blood or from apheresis) and by performing the necessary testing to ensure that the product meets the minimum requirements for the named products, as indicated in Table 1.]
Table 1. Red Blood Cells Solutions Product Name Red Blood Cells Red Blood Cells, Pheresis Red Blood Cells, Leukocyte Reduced Red Blood Cells, Frozen Method of Solution Prepared from whole human blood Prepared using automated apheresis systems Prepared from Red Blood Cells or Red Blood Cells, Pheresis (total leukocytes count <5 106) Prepared from Red Blood Cells or Red Blood Cells, Pheresis suspended in cryoprotective solution (glycerol) and frozen at an appropriate temperature Prepared from Red Blood Cells, Frozen, from which glycerol is removed by washing by an approved procedure
Red Blood Cells, Deglycerolyzed
Label it to indicate, the ABO Group/Rh Type, as indicated in Table 2. [NOTEEvery Red Blood Cell product must have a determination made as to its ABO Group and Rh Type and specificity of unexpected red cell antibodies, if any] If an ABO blood group color scheme is used, use the following labeling color: Group A (yellow), Group B (pink), Group O (blue), and Group AB (white). Label the Red Blood Cells with names of the adventitious agents tested and the results of the tests. If it has been issued prior to determination of the test results, label it also
AS
contains sodium chloride, dextrose, adenine, and other substances that support red cell survival and function. Examples of such solutions are AS-1, AS-3, and AS-5.
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with a warning Donor Untested and to specify Uncrossmatched Blood, when appropriate.
Table 2. Blood Group and Rh Type ABO Group A A B B AB AB O O Rh Type Positive Negative Positive Negative Positive Negative Positive Negative
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Analysis: Place a small drop of the diluted platelets onto the end of a clean glass microscope slide. Obtain a second clean glass microscope slide, and draw its edge across the drop of platelets so that capillary action spreads the drop across the first slide. Push the second slide in one smooth motion across the first slide to make a smear of platelets. Allow the smear to dry. Apply a liberal amount (1 to 2 mL) of Wrights stain to the platelet smear, and allow to stand for 2 min. Dip the slide in deionized water, and gently blot dry with absorbent paper. Examine the slide using a microscope at 100 with bright illumination. Acceptance criteria: Platelets appear as anuclear round or oval shapes approximately 0.1 to 0.2 m in diameter, with some fine purple granulation. The platelets may occasionally appear to have a purple center with clear cytoplasm at the periphery SPECIFIC TESTS PLATELET COUNT: Use a commercially available, validated hematology analyzer to determine platelet count. Proceed as directed in the instruments operating manual RESIDUAL LEUKOCYTE COUNT: Sample solution: Transfer 100 L of Platelets into a suitable test tube and add 400 L of 0.01% (w/v) crystal violet in 15% (v/v) acetic acid, and mix thoroughly. Analysis: Fit a hemocytometer with a 50-L counting volume and a bright background with a cover slip. Load the counting chamber with the mixture until the counting area is completely covered, but do not overfill. Cover the counting chamber with a suitable moist lid to prevent evaporation, and allow to settle undisturbed for 10 to 15 min. Remove the lid, and place the chamber on the stage of a light microscope fitted with a 10 ocular lens and 20 objective. Count the leukocytes in the entire 50-L counting volume. Calculate the leukocyte count in the platelets, expressed in leukocytes/L, by dividing the observed leukocyte count by 10. Calculate the total number of leukocytes in the red blood cell unit. Total leukocytes = leukocytes/L 103 volume of the platelet unit in mL PH 791 (for stored Platelets): Transfer aseptically a volume appropriate for the equipment: the pH must be greater than 6.2 throughout the storage period. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Store Platelets in an approved container. Platelets may be stored in plasma or in an approved additive solution at 20 to 24 with continuous gentle agitation for no more than 5 days after date of preparation. LABELING: Label the container to indicate the collection date, the donation number or other coding means to uniquely identify the unit and to provide traceability to the donor, approximate volume, storage temperature, and its expiration date. Indicate the anticoagulant solution used and any additive solutions added subsequent to collection. Also label the container to identify donor status (for example, volunteer or paid). Label it also with the following statements: See Circular of Information for the Use of Human Blood and Blood Components for indications, contraindications, cautions, and methods of infusion; Properly identify intended recipient; This product may transmit infectious agents; and Rx only. In addition, label the container to indicate the product name as indicated in Table 1. [NOTEThe name is determined by the method of platelets solution (derived from whole blood or by apheresis) and by performing the necessary testing to ensure that the product meets the minimum requirements for the named products, as indicated in Table 1.].
USP REFERENCE STANDARDS 11 USP Hemoglobin RS
Platelets
(Comment on this Monograph)id=m763=Platelets=BMonos.pdf) DEFINITION Platelets is the portion of blood that contains platelet cells. It is derived from human whole blood from which red blood cells and a portion of the plasma are removed by centrifugation, sedimentation, or apheresis. In the apheresis removal method, the red blood cells and plasma are automatically removed and returned directly to the donor. Platelets derived from whole blood must be prepared within 4 h after collecting the whole blood from which it is derived, or within the time frame specified for the blood collecting, processing, and storage system used. Platelets may be derived from whole blood collected in any approved anticoagulant solution (see USP monographs for anticoagulant solutions). Platelets prepared by apheresis must be collected using Anticoagulant Citrate Dextrose Solution A as the anticoagulant solution. Platelets derived from whole blood should have a minimum of 5.5 1010 platelet cells suspended in a volume of 40 to 70 mL of original plasma. Platelets produced by apheresis should have a minimum of 3.0 1011 platelet cells, suspended in 100 to 500 mL of original plasma or in an approved additive solution. Platelets derived from whole blood or by apheresis may be further processed by filtration for removal of leukocytes, or by irradiation to inactivate lymphocytes. Platelets derived from whole blood may be pooled from multiple donors to form one dose of platelets. Leukocytes may be removed from platelets by filtration, using an approved platelet leukoreduction filter. Platelets derived from whole blood must contain less than 8.3 105 leukocytes after filtration. The source blood for platelets must be tested for syphilis, hepatitis B, and human T-cell Lymphotropic Virus (HTLV) Type I and Type II, using FDA-approved and -licensed commercially available test kits. The test results must be below the limits of detection specified by the manufacturers of the respective test kits. The source blood must also be tested for hepatitis C and HIV Type 1 and Type 2, using FDA-approved nucleic acid assays. The test results must be below the approved limits of detection for the tests used. IDENTIFICATION PROCEDURE Sample solution: Dilute a small volume of Platelets 1:1000 with 0.9% sodium chloride solution
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Table 1. Names of Platelet Solutions Product Name Platelets Platelets, Pooled Method of Solution Prepared from a single unit of whole human blood within 8 h of collection. Individual platelet units derived from whole human blood and pooled by aseptic techniques. [NOTELabel this solution
Official Monographs / Bovine 115

plastic, and other reconstructive procedures to contribute to the repair, reinforcement, and generation of tissue. The sterile material is surgically secured, onlayed, and/or packed into deficient soft tissues such as skin, tendon, muscle, and dura mater. The source fetal or neonatal bovine skin is mechanically and chemically processed to isolate the dermis and remove cells and cellular components. To prevent the transmission of infectious disease, the manufacturing process has been validated to inactivate viruses potentially present in the source material. To prevent the spread of transmissible spongiform encephalopathies, the source material is acquired from appropriate geographic locations in accordance with relevant guidelines subject to governmental oversight. The product is inspected and tested to assure the product meets specifications. IMPURITIES Inorganic Impurities ASH DETERMINATION Analysis: Place a sample of the final product, 5.0 g, in a kiln-dried, porcelain crucible. Record the weight to the nearest 0.1 mg. Place the crucible containing the sample into an oven at 125 for 24 h. Then place the crucible containing the sample into a cool muffle furnace. Heat the furnace to 350, and maintain the temperature until smoking ceases (generally about 20 min). Heat the furnace to 550. Maintain the temperature for 2 h. Cool the crucible in a desiccator. Weigh the crucible, and record the weight to the nearest 0.1 mg. Calculate the percentage of ash: Result = [(W1 W2)/W3] 100 = weight of crucible and residue (g) W1 = weight of crucible (g) W2 W3 = weight of sample (g) Acceptance criteria: NMT 0.3% SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements BACTERIAL ENDOTOXINS TEST 85: It meets the requirements as directed under Transfusion and Infusion Assemblies and Similar Medical Devices 161. HISTOLOGICAL EVALUATION 1% Acid alcohol: 70% ethyl alcohol and hydrochloric acid (37.5%) (99:1) Potassium alum solution: 100 mg/mL of potassium alum in distilled water, dissolve with the aid of heat and a magnetic stirrer Hematoxylinalcohol solution: 100 mg/mL of hematoxylin (see Reagents, Indicators, and SolutionsReagent Specifications) in 100% ethyl alcohol, dissolve at room temperature Hematoxylin solution: Slowly combine the 1000 mL of the Potassium alum solution with 50 mL of the Hematoxylinalcohol solution. Bring to a boil as rapidly as possible. Remove from heat and slowly add 2.5 g of mercuric oxide. Return the solution to heat until it becomes dark purple, remove from heat, and cool in a sink of cold water. Eosin solution: Dissolve 1.0 g of eosin Y, water soluble, in 100 mL of distilled water. Dissolve 1.0 g of phloxine B in 100.0 mL of distilled water. Combine 100 mL of eosin Y solution with 10 mL of phloxine B solution, 780 mL of 95% ethyl alcohol, and 4.0 mL of glacial acetic acid. [NOTEFilter daily before use.] Bluing agent: 15.4 mg/mL of lithium carbonate in distilled water 10% Neutral buffered formalin: To 6.5 g of dibasic sodium phosphate (anhydrous) and 4.0 g of monobasic
with a unique identifying number related to the number of individual units pooled, and with an expiration date of 4 h after pooling of the individual units.]
Platelets, Pheresis Platelets, Leukocyte Reduced
Prepared by apheresis from a single donor. Prepared from whole blood, either by centrifugation or by sedimentation, and filtered using an approved platelet leukoreduction filter to yield less than 8.3 105 white blood cells in the final container. Contains less than 5 106 white blood cells, prepared by apheresis, with or without a filter.
Platelets, Pheresis, Leukocyte Reduced
[NOTEPlatelets prepared by apheresis should be labeled with the donors ABO blood group and Rh factors. Test the donors whole blood or red blood cells as directed under Whole Blood or Red Blood Cells, respectively.]
Botulism Antitoxin
(Comment on this Monograph)id=m10120=Botulism Antitoxin=B-Monos.pdf) DEFINITION Botulism Antitoxin conforms to the regulations of the federal Food and Drug Administration concerning biologics (See Biologics 1041). It is a sterile, nonpyrogenic solution of the refined and concentrated antitoxic antibodies, chiefly globulins, obtained from the blood of healthy horses that have been immunized against the toxins produced by the type A and type B and/or type E strains of Clostridium botulinum. Its potency is determined with the U.S. Standard Botulism Antitoxin of the relevant type, tested by neutralizing activity in mice of the corresponding U.S. Control Botulism Test Toxin. It contains NMT 20.0% of solids, and contains a suitable antimicrobial agent. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers only, at a temperature between 28. EXPIRATION DATE: The expiration date for Antitoxin containing a 20% excess of potency is not later than 5 years after date of issue from manufacturers cold storage (5, 1 year; or 0, 2 years). LABELING: Label it to state that it was prepared from horse blood.
Bovine Acellular Dermal Matrix

(Comment on this Monograph)id=m1676=Bovine Acellular Dermal Matrix=B-Monos.pdf) DEFINITION Bovine Acellular Dermal Matrix is a remodelable collagen scaffold derived from fetal or neonatal bovine skin. It is presented to the physician as a flat white sheet that is cut to size and hydrated in room temperature sterile saline solution prior to implantation. It is utilized as a structural scaffold in orthopedic, neurosurgical, urogynecological, dermatological,
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Bovine / Official Monographs
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LIPID ANALYSIS Analysis: A standard Soxhlet extraction apparatus is required. Dry flasks in an oven/dessicator and weigh, recording the weight to the nearest 0.0001 g. Grind or cut into small pieces 3.04.0 g of test material and place into a thimble. Record the weight of the test material to the nearest 0.0001 g. Place the thimble of material and 8090 mL of petroleum ether into an extraction flask, and place into the Soxhlet extraction tube. Reflux for 4 h. Collect all of the ether into the flask, and evaporate. Weigh the flask, recording weight to the nearest 0.0001 g. Calculation: For the weight of lipid, substract the weight of the clean flask from the final weight of the flask. Calculate the percent of lipid based on the weight of the starting material. Acceptance criteria: The percentage of lipid in 3.04.0 g of Dermal Matrix sample is between 0% and 1.5%. MOISTURE CONTENT Analysis: Proceed as directed under Loss on Drying 731 to calculate the moisture content, with the following specifics. Mince approximately 5.0 g of Dermal Matrix; place it into an aluminum dish. Dry the sample in an air oven for 1618 h at 100102. Calculate the percentage of moisture in the sample taken: Result 1 = [(W1 W2)/W3] 100 W1 W2 W3 = weight of dried sample and pan (g) = weight of pan (g) = weight of sample (g) Result 2 = 100 Result 1 Acceptance criteria: 10.0%12.0% of the original sample weight CARBOHYDRATES Analysis: The percentage of carbohydrates is determined: Carbohydrate% = 100% (lipid% + protein% + moisture% + ash%) Acceptance criteria: The percentage of carbohydrates is equal to or less than 0.0%. Because this is a calculated value, influenced by the error inherent in the test methods above (Lipid analysis, Moisture content, and Ash determination), a calculated value less than 0.0% is acceptable. GEL ELECTROPHORESIS: Use the electrophoresis determination method as directed under Biotechnology-Derived Articles Polyacrylamide Gel Electrophoresis 1056 with the following specifics. Collagen extraction solution: Prepare a 0.5M acetic acid solution containing 2 mM ethylenediaminetetraacetic acid (EDTA). 2X Tris-glycine sample buffer: Prepare a 2X solution containing 63 mM Tris-HCl pH 6.8, 10% glycerol, 2% sodium dodecyl sulfate (SDS), 0.05% 2-mercaptoethanol, and 0.25% bromophenol blue4. 1X Sample buffer: 2X Tris-glycine sample buffer and water (1: 1) SDS-PAGE running buffer: Prepare a solution containing 25 mM Tris pH 8.3, 192 mM glycine, and 0.1% SDS5. Polyacrylamide gel: Prepare a Tris-HCl polyacrylamide gel with a 4% to 20% gradient6.
suitable sample buffer can be obtained from Invitrogen Corporation, 1600 Faraday Ave., P.O. Box 6482, Carlsbad, CA 92008. 5A suitable gel running buffer can be obtained from Bio-Rad Laboratories, 1000 Alfred Nobel Dr., Hercules, CA 94547. 6A suitable precast acrylamide gel can be obtained from Bio-Rad Laboratories, 1000 Alfred Nobel Dr., Hercules, CA 94547.
4A
sodium phosphate, add 900 mL of distilled water and 100 mL of formaldehyde (37%40%). Sample solution and staining: Remove a sample of finished product with an 8.0-mm biopsy punch. Place the sample in a labeled tissue cassette, and fix for 24 h in 10% Neutral buffered formalin. Dehydrate the sample in sequential soaks of the following: 70% ethyl alcohol (45 min), 80% ethyl alcohol (45 min), 95% ethyl alcohol (90 min), 100% ethyl alcohol (180 min), and xylene (90 min). Embed the sample in melted paraffin, cool, and cut 5-m thick sections with a microtome. Collect sections on microscope slides. Deparaffinize the slide with xylene and hydrate with distilled water. Stain in Hematoxylin solution for 615 min. Wash in running tap water for 25 min. Dip two times in 1% Acid alcohol. Wash briefly in tap water. Place in Bluing agent until the sections are bright blue. Wash in running tap water for 10 min. Place in 80% ethyl alcohol for 12 min. Dehydrate and clear through two changes each of 95% ethyl alcohol, 100% ethyl alcohol, and xylene, 2 min each. Affix a coverslip over the tissue using an appropriate resinous mounting media. The nuclei stains blue, the cytoplasm stains from pink to red, and the collagen fibers stain from pink to red. Microscopic and morphological characteristics: The collagen fibers of the Dermal Matrix stain pink-red, and no evidence of cell nuclei or cytoplasm are apparent in prepared histological sections as shown in the USP Bovine Acellular Dermal Matrix Reference Photomicrographs of products with acceptable histological appearance. PROTEIN DETERMINATION: Use the Kjeldahl nitrogen (protein) determination method to calculate the percent protein of the final product as directed under Nitrogen Determination 461 with the following specifics. Suitable equipment and procedures are readily available1. Digestion: Prepare a rack of 1520 Kjeldahl digestion tubes. In each, place 2.02.2 g of final product, 0.2 0.05 g of ammonium sulfate, a metallic catalyst tablet,2 and boiling chips.3 Prepare a blank tube with catalyst tablets and boiling chips (reagent blank). To each tube add 15 mL of concentrated sulfuric acid, and then, very slowly, 3 mL of hydrogen peroxide (30%35%). Place the digestion tubes on a digestion block, and heat to 410. Digest for 60 5 min. The mixture in the tubes should be a clear green. Distillation: Add excess base (50% sodium hydroxide). Generally, for each 5 mL of concentrated sulfuric acid used in the digestion, 20 mL of 40% (w/w) sodium hydroxide is required to make the digest strongly alkaline (pH > 11). Mix each tube and let cool to room temperature. Distill each tube to collect approximately 125 mL of total distillate in a flask containing 25 mL of 4% boric acid. A reagent blank is run with each set. Titration: Titrate the collected distillate with standardized 0.2 N sulfuric acid to a neutral gray color endpoint. Record the volume of sulfuric acid used. Analysis: Calculate the percentage of protein: Result = [(S1 B) S2 W1 P]/W2 = sulfuric acid (mL) S1 B = blank (mL) = N of sulfuric acid S2 = milliequivalent weight N 100 (%), 1.4007 W1 P = protein factor for meat, 6.25 = weight of sample (g) W2 Acceptance criteria: The percentage of protein in 2.02.2 g of Dermal Matrix sample is between 90.0% and 95.0%.
1A
suitable device and associated procedures can be obtained from Labconoco, 8811 Prospect Ave., Kansas City, MO. 2A suitable catalyst is Pro-Pac CT-37, Alfie Packers, 8901 J St., Omaha, NE. 3Commonly referred to as Henger granules.
USP 32
Molecular weight marker: Use a suitable molecular weight marker containing protein bands between 10 and 250 kilodaltons (kDa). Staining solution: Prepare a solution containing 0.25% (w/v) Coomassie brilliant blue R-250 (see Reagents, Indicators, and SolutionsReagent Specifications) in 10% acetic acid and 10% npropanol. Destain solution: Water, acetic acid, and npropanol (8:1:1) Collagen preparations: Mince 0.5 g of Dermal Matrix final product. Weigh a sample of minced Dermal Matrix, and add to a volume of Collagen extraction solution to obtain a concentration of 5 mg/mL (dry weight of Dermal Matrix). Extract on a rocking platform at room temperature for 72 h. Analysis: Dilute acid-extracted collagen samples in 2X Trisglycine sample buffer to a concentration of 0.5 mg/mL, and incubate for 5 min at 100. Load the Polyacrylamide gel in the electrophoresis apparatus, and add SDS-PAGE running buffer to the top and bottom reservoirs. Load 10 L of Molecular weight markers in the first well of the Polyacrylamide gel and 10 L of 1X Sample buffer in the second well. Load 10 L (5 g) of each collagen sample into subsequent gel wells. Attach the cathode and anode to the appropriate terminals, and apply 110 V to the gel. Run the gel until the bromophenol blue reaches the bottom of the gel. Remove the gel from the electrophoresis apparatus, and place it in a tray containing enough Staining solution to cover the gel. Incubate the gel for 3 h on a rocker at room temperature. Completely remove the Staining solution from the tray, cover the gel with Destain solution, and slowly rock the gel for 20 min. Remove the Destain solution, and repeat the destaining procedure three times. Inspect the gel for bands that have migrated from the origin. System suitability: All bands between 20 and 200 kDa are present. The lane containing 1X Sample buffer does not contain any bands. Data analysis: Where a protein band appears in the gel, the molecular weight of this protein is determined by comparing the position of the band to that of the known Molecular weight marker. Specificity: The lanes of the Polyacrylamide gel that correspond to Dermal Matrix show four major protein bands. Two bands, when compared to the Molecular weight marker, appear at 96 and 94 kDa. These two bands correspond to the monomeric alpha 1 and alpha 2 chains of collagen Type I, respectively. Another two bands appear close together at 200 kDa, which correspond to alpha 1 and alpha 1/alpha 2 collagen dimers. TENSILE STRENGTH Analysis: Cut samples 5-mm wide 50-mm long from representative pieces from final product lots. Measure the thickness of the sample. Test the samples with a commercially available material test system.7 Mount and align the specimen, gripping 1 cm of the sample on both ends to ensure a sample gauge length of 3 cm. Pull the grips apart at 30 mm/min while concurrently measuring the force exerted on the sample. Record the maximum force (N) measured during the test. Calculate the tensile strength (n/mm2): Result = F/W T F W
7A
Official Monographs / Bovine 117

T = thickness (mm) Acceptance criteria: The measured tensile strength for each lot is NLT 5 N/mm2. SUTURE RETENTION FORCE Analysis: Cut representative 1 1 cm samples from final product lots. Using an appropriate suture material (e.g., 40 polypropylene suture), thread the suture 3 mm from the edge of the sample in the center and pull through. Clamp approximately 5 mm of the opposite, unsutured end of the sample in the upper pneumatic grip of a commercially available material test system.8 The suture tails are hanging freely. Clamp the suture tails to the lower grip. Pull the grips apart at 20 mm/min while concurrently measuring the force exerted. Record the maximum force (N) measured. Acceptance criteria: The suture retention force measured for each lot is NLT 5N for a 1 1 cm sample of the Dermal Matrix. THERMAL ANALYSIS Analysis: A final product sample of approximately 1020 mg is heated at 2/min from 3090, hydrated with water, and the thermal characteristics of each processed skin is measured with a differential scanning calorimeter as directed under Thermal Analysis 891. Dermal Matrix displays a single thermal transition peak between 58 and 67. VISUAL INSPECTION Analysis: Each piece of final product is visually inspected under a white light at a distance of 3045 cm for color, the presence of particulates, and holes. Dermal Matrix is white, and neither particulates nor holes are visible. HYDRATION RATE Analysis: Cut a sample of finished product lot approximately 1 1 cm. The sample fully hydrates, as indicated by a change in color from white to gray, in less than 3 min when placed in room temperature saline solution.
= maximum force (N) = width, 5 mm
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: The package is a sealed, foil pouch that provides an effective moisture, light, gas, and sterility barrier. Store in clean, dry conditions between 15 and 30. LABELING: Label it to indicate that it is derived from bovine origin. The product is labeled to indicate the products intended clinical use. It is labeled with the dimensions of the product, the expiration date, the required storage conditions, lot number, part number, and the manufacturers name and address. The label indicates that the product is sterile and nonpyrogenic and is designed for single patient, one-time use. The labeling cautions the user to inspect the packaging for damage and to discard the product if the packaging has been compromised. The labeling also cautions the user to hydrate the product only in room temperature sterile saline solution. USP REFERENCE STANDARDS 11 USP Endotoxin RS USP AUTHENTIC VISUAL REFERENCES 11 USP Bovine Acellular Dermal Matrix Reference Photomicrographs. These Photomicrographs show the histological appearance of failed, cell-containing source material (Photomicrographs 1 and 2) and of passing, processed, decellularized material (Photomicrographs 3 and 4). The samples were prepared as directed in the test for Histological evaluation.
8A
suitable material test system is available from Instron Corporation, 825 University Ave., Norwood, MA.
suitable material test system is available from Instron Corporation, 825 University Ave., Norwood, MA.
118
Bretylium / Official Monographs
USP 32
from the Sample solution is NMT two times the bretylium response from the Standard solution (2%); and no individual peak response is greater than that of the bretylium peak from the Standard solution (1%). SPECIFIC TESTS LOSS ON DRYING 731: Dry it in a vacuum at 75 for 2 h: it loses NMT 3.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Bretylium Tosylate RS
Bretylium Tosylate
(Comment on this Monograph)id=m10170=Bretylium Tosylate=B-Monos.pdf)
414.36 C18H24BrNO3S Benzenemethanaminium, 2-bromo-N-ethyl-N,N-dimethyl-, salt with 4-methylbenzenesulfonic acid (1:1); (o-Bromobenzyl)ethyldimethylammonium p-toluenesulfonate [61-75-6; 59-41-6]. DEFINITION Bretylium Tosylate contains NLT 98.0% and NMT 101.0% of C18H24BrNO3S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Organic ImpuritiesProcedure. ASSAY PROCEDURE Sample solution: 6 mg/mL of Bretylium Tosylate in dioxane Titrant: 0.025 N perchloric acid in dioxane, standardized as described in perchloric acid, tenth-normal in dioxane VS Analysis: To 50 mL of Sample solution, add 2 drops of crystal violet TS, and titrate with Titrant to a blue-green endpoint. Perform a blank determination (see Titrimetry 541), and make any necessary correction. Each mL of Titrant is equivalent to 10.36 mg of C18H24BrNO3S. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method I 231: NMT 20 ppm Organic Impurities PROCEDURE Solution A: 0.01 M 1-sodium octanesulfonate Mobile phase: Acetonitrile, glacial acetic acid, triethylamine, and Solution A (19:2:0.5:81) Standard solution: 0.02 mg/mL of USP Bretylium Tosylate RS in Mobile phase Sample solution: 2 mg/mL of Bretylium Tosylate in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 4.6-mm 25-cm; packing L11 Flow rate: 1.9 mL/min Injection size: 30 L System suitability Sample: Standard solution [NOTEThe relative retention times for tosylate ion, o-bromobenzyldimethylamine, bretylium, m-bromobenzyldimethylamine, and pbromobenzyldimethylamine are 0.25, 0.74, 1.0, 1.27, and 1.40, respectively.] Suitability requirements Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Acceptance criteria: The sum of the responses for all the peaks, excluding those of the bretylium and tosylate peaks,
Bretylium Tosylate Injection

(Comment on this Monograph)id=m10180=Bretylium Tosylate Injection=B-Monos.pdf) DEFINITION Bretylium Tosylate Injection is a sterile solution of Bretylium Tosylate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C18H24BrNO3S. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.38 mg of monobasic sodium phosphate and 2.0 mL of 25% tetra-methylammonium hydroxide solution in methanol in 800 mL of water, adjust with phosphoric acid to a pH of 3.1 0.1, dilute with water to 1000 mL Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A (15:3:182) Standard solution: 0.2 mg/mL of USP Bretylium Tosylate RS Sample solution: Equivalent to 0.2 mg/mL of bretylium tosylate from a volume of Injection in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for tosylate and bretylium are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0, between the bretylium and tosylate peaks Relative standard deviation: NMT 1.4% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H24BrNO3S in each mL of the Injection: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response of bretylium in the Sample solution = peak response of bretylium in the Standard solution = concentration of USP Bretylium Tosylate RS in the Standard solution (mg/mL)
USP 32
= concentration of bretylium tosylate taken to prepare the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Unit/mg of bretylium tosylate PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 3.57.0 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Bretylium Tosylate RS USP Endotoxin RS CU
Official Monographs / Bretylium 119

Suitability requirements Resolution: NLT 3.0, between the bretylium and tosylate peaks Relative standard deviation: NMT 1.4% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H24BrNO3S in each mL of the Injection: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response of bretylium from the Sample solution = peak response of bretylium from the Standard solution = concentration of USP Bretylium Tosylate RS in the Standard solution (mg/mL) = concentration of dextrose injection taken to prepare the Sample solution (mg/mL)
Bretylium Tosylate in Dextrose Injection

(Comment on this Monograph)id=m10190=Bretylium Tosylate in Dextrose Injection=B-Monos.pdf) DEFINITION Bretylium Tosylate in Dextrose Injection is a sterile solution of Bretylium Tosylate and Dextrose in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amounts of bretylium tosylate (C18H24BrNO3S) and dextrose (C6H12O6 H2O). It contains no antimicrobial agents. IDENTIFICATION A. The retention time of the major peak in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay, Bretylium tosylate B. Add a few drops of a solution (1 in 20) to 5 mL of hot alkaline cupric tartrate TS: a copious red precipitate of cuprous oxide is formed. ASSAY BRETYLIUM TOSYLATE Solution A: 1.38 mg of monobasic sodium phosphate and 2.0 mL of 25% tetra-methylammonium hydroxide solution in methanol in 800 mL of water. Adjust with phosphoric acid to a pH of 3.1 0.1, dilute with water to 1000 mL. Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A (15:3:182) Standard solution: 0.2 mg/mL of USP Bretylium Tosylate RS Sample solution: Equivalent to 0.2 mg/mL of bretylium tosylate from a volume of Injection in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for tosylate and bretylium are about 0.7 and 1.0, respectively.]
DEXTROSE Sample solution: Equivalent to 25 g of dextrose of the Dextrose Injection, to a 100-mL volumetric flask. Add 0.2 mL of 6 N ammonium hydroxide, and dilute with water to volume. Analysis: Determine the angular rotation in a suitable polarimeter tube (see Optical Rotation 781). Calculate the percentage (g/100 mL) of C6H12O6 H2O in the portion of Injection: (Mr1/Mr2) A R (100/Sr) = molecular weight for dextrose monohydrate, 198.17 Mr2 = molecular weight for anhydrous dextrose, 180.16 A = 100 mm divided by the length of the polarimeter tube (mm) R = observed rotation (degrees) = midpoint of the specific rotation range for Sr anhydrous dextrose (degrees), 52.9 Acceptance criteria: 95.0%105.0% Mr1 SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Unit/mg of bretylium tosylate PH 791: 3.06.5 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose glass or plastic containers. Glass containers are preferably of Type I or Type II glass. USP REFERENCE STANDARDS 11 USP Bretylium Tosylate RS USP Endotoxin RS
120
Brinzolamide / Official Monographs

CU
USP 32
= nominal concentration of Brinzolamide in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1 Mobile phase: Dehydrated alcohol, chromatographic solvent hexane, methanol, and diethylamine (55:40:5:0.2) System suitability solution: 0.4 mg/mL of USP Brinzolamide RS and 0.02 mg/mL of USP Brinzolamide Related Compound A RS in dehydrated alcohol Sample solution: 0.5 mg/mL of Brinzolamide in dehydrated alcohol Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L51 Flow rate: 0.75 mL/min Injection size: 5 L System suitability Sample: System suitability solution [NOTEThe relative retention times for brinzolamide and brinzolamide related compound A are 1.0 and 1.2, respectively.] Suitability requirements Resolution: NLT 1.8 between brinzolamide and brinzolamide related compound A Column efficiency: NLT 2000 theoretical plates from brinzolamide Tailing factor: NMT 1.8 for the brinzolamide peak Analysis Sample: Sample solution Calculate the percentage of brinzolamide related compound A in the portion of Brinzolamide taken: Result = (rU/rS) 100 = peak response for brinzolamide related compound A = sum of the peak responses for brinzolamide and rS brinzolamide related compound A Acceptance criteria: NMT 0.5% of brinzolamide related compound A is found PROCEDURE 2 Buffer: Prepare as directed in the Assay. Mobile phase A: Prepare as directed for Mobile phase in the Assay. Mobile phase B: Acetonitrile and Buffer (7:13) System suitability solution: 0.1 mg/mL of each of USP Brinzolamide RS and USP Brinzolamide Related Compound B RS in Mobile phase A Sample solution: 1 mg/mL of Brinzolamide in Mobile phase A Chromatographic system (See Chromatography 621, System Suitability.) rU
Brinzolamide
(Comment on this Monograph)id=m10206=Brinzolamide=BMonos.pdf)
C12H21N3O5S3 383.51 2H-Thieno[3,2-e]-1,2-thiazine-6-sulfonamide, 4(ethylamino)-3,4-dihydro-2-(3-methoxypropyl)-, 1,1-dioxide, (R)-; (R)-4-(Ethylamino)-3,4-dihydro-2-(3-methoxypropyl)-2H-thieno [3,2-e]-1,2-thiazine-6-sulfonamide 1,1-dioxide [138890-62-7]. DEFINITION Brinzolamide contains NLT 98.0% and NMT 102.0% of C12H21N3O5S3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the System suitability solution, as obtained in Organic Impurities, Procedure 1. ASSAY PROCEDURE Buffer: Add 4.0 mL of triethylamine to 1000 mL of water, and adjust with phosphoric acid to a pH of 3.0. Mobile phase: Acetonitrile and Buffer (1:3) Standard solution: 0.1 mg/mL of USP Brinzolamide RS in Mobile phase Sample solution: 0.1 mg/mL of Brinzolamide in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1200 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H21N3O5S3 in the portion of Brinzolamide taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Brinzolamide RS in the Standard solution (mg/mL)
USP 32
Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution Equilibrate the system with Mobile phase A. [NOTEThe relative retention times for brinzolamide related compound B and brinzolamide are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between brinzolamide and brinzolamide related compound B Column efficiency: NLT 1200 theoretical plates from brinzolamide Tailing factor: NMT 2.0 for the brinzolamide peak Analysis A Samples: Mobile phase A and Sample solution Allow the elution to continue for 20 min, and measure the areas for all the peaks, excluding the peaks of Mobile phase A. Calculate the percentage of each impurity in the portion of Brinzolamide taken: Result = (ri/rS) 100 ri = peak response for each impurity = sum of the responses for all the peaks rS Acceptance criteria: NMT 0.3% of any individual impurity Analysis B Equilibrate the system with Mobile phase B. Samples: Sample solution Again, record the chromatograms, allowing the elution to continue for 20 min, and measure the areas for brinzolamide and all the peaks having a relative retention time greater than 6. Calculate the percentage of each impurity in the portion of Brinzolamide taken: Result = (ri/rS) 100 = peak response for each impurity ri = sum of the responses for all the peaks rS Acceptance criteria: NMT 0.3% of any individual impurity; NMT 1.0% of total impurities from Analysis A and Analysis B SPECIFIC TESTS LOSS ON DRYING 731: Dry in vacuum at 100105 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Brinzolamide RS USP Brinzolamide Related Compound A RS USP Brinzolamide Related Compound B RS
Official Monographs / Brinzolamide 121

IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: 11.75 g/L of ammonium acetate in water, adjust with acetic acid to a pH of 5.2 Mobile phase: Methanol and Buffer (7:13) Standard solution: 0.2 mg/mL of USP Brinzolamide RS in Mobile phase Sample solution: Transfer a volume of Ophthalmic Suspension, equivalent to about 10 mg of brinzolamide, to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Nominal concentration 0.2 mg/mL System suitability solution: 0.06 mg/mL of USP Brinzolamide Related Compound B RS in Standard solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for brinzolamide related compound B and brinzolamide are about 0.48 and 0.61, respectively.] Suitability requirements Resolution: NLT 4.5 between the brinzolamide and brinzolamide related compound B, System suitability solution Column efficiency: NLT 2500 theoretical plates, System suitability solution Tailing factor: NMT 2.0, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H21N3O5S3 in the portion of Ophthalmic Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Brinzolamide RS in the Standard solution (mg/mL) CU = concentration of Brinzolamide in the Sample solution Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE 1 Mobile phase: Dehydrated alcohol, hexane, methanol, and diethylamine (55:40:5:0.2) System suitability solution: 0.4 mg/mL of USP Brinzolamide RS and 0.02 mg/mL of USP Brinzolamide Related Compound A RS in dehydrated alcohol Sample solution: Transfer volume of Ophthalmic Suspension, equivalent to about 10 mg of brinzolamide, to a 25-mL volumetric flask. Dilute with dehydrated alcohol to volume. Nominal concentration 0.2 mg/mL Chromatographic system (See Chromatography 621, System Suitability.) rU rS CS
Brinzolamide Ophthalmic Suspension

(Comment on this Monograph)id=m10208=Brinzolamide Ophthalmic Suspension=B-Monos.pdf) DEFINITION Brinzolamide Ophthalmic Suspension is a sterile, aqueous suspension of Brinzolamide containing a suitable antimicrobial preservative. It contains NLT 90.0% and NMT 110.0% of the labeled amount of brinzolamide (C12H21N3O5S3).
122
Brinzolamide / Official Monographs

PH 791: 6.58.5
USP 32
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L51 Flow rate: 0.75 mL/min Injection size: 5 L System suitability Sample: System suitability solution [NOTEThe relative retention times for brinzolamide and brinzolamide related compound A are about 1.0 and 1.2, respectively.] Suitability requirements Resolution: NLT 1.8 between brinzolamide and brinzolamide related compound A Column efficiency: NLT 2000 theoretical plates from brinzolamide Tailing factor: NMT 1.8 for the brinzolamide peak Analysis Sample: Sample solution Calculate the percentage of brinzolamide related compound A in the portion of Brinzolamide taken: Result = (rU/rS) 100 = peak response of brinzolamide related compound A in the Sample solution = sum of the peak responses for brinzolamide and rS brinzolamide related compound A in the Sample solution Acceptance criteria: NMT 1.5% of brinzolamide related compound A is found PROCEDURE 2 Buffer and Mobile phase: Proceed as directed in the Assay. Standard solution: 2.5 g/mL of USP Brinzolamide Related Compound B RS in Mobile phase Sample solution: Use the Sample solution as prepared in the Assay. Chromatographic system: Proceed as directed in the Assay. System suitability Proceed as directed in Assay. Analysis: Proceed as directed in the Assay Calculate the percentage of each impurity in the portion of Ophthalmic Suspension taken: rU Result = (rU/rS) (CS/CU) (Mr1/Mr2) F 100 = peak response for each impurity from the Sample solution rS = peak response for USP Brinzolamide Related Compound B RS from the Standard solution CS = concentration of USP Brinzolamide Related Compound B RS in the Standard solution (mg/mL) = nominal concentration of brinzolamide in the CU Sample solution (mg/mL) = molecular weight of des-ethyl brinzolamide, Mr1 356.46 = molecular weight of des-ethyl brinzolamide Mr2 oxalate, 445.49 F = unit conversion factor, 0.001 mg/g Acceptance criteria: NMT 0.5% of any individual impurity is found Total impurities: NMT 2.0% rU SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at a temperature between 4 and 30. USP REFERENCE STANDARDS 11 USP Brinzolamide RS USP Brinzolamide Related Compound A RS USP Brinzolamide Related Compound B RS
Bromocriptine Mesylate
(Comment on this Monograph)id=m10230=Bromocriptine Mesylate=B-Monos.pdf)
C32H40BrN5O5 CH4SO3 Ergotaman-3,6,18-trione, 2-bromo-12-hydroxy-; 2-(1-methylethyl)-5-(2-methylpropyl)-; monomethanesulfonate (salt), (5)-; 2-Bromoergocryptine monomethanesulfonate (salt) [22260-51-1].
750.70
DEFINITION Bromocriptine Mesylate contains NLT 98.0% and NMT 102.0% of C32H40BrN5 O5 CH4SO3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M: Undried B. ULTRAVIOLET ABSORPTION 197U Solution: 50 g/mL in 0.1 M methanolic methanesulfonic acid ASSAY PROCEDURE Sample solution: 600 mg of Bromocriptine Mesylate into a titration vessel, dissolve in 80 mL of a mixture of acetic anhydride and glacial acetic acid (7:1) Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 75.07 mg of C32H40BrN5O5 CH4SO3. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1: LIMIT OF METHANESULFONIC ACID CONTENT: NLT 12.5% and NMT 13.4% of CH3SO3H Sample solution: 400 mg into a titration vessel, dissolve in 70 mL of methanol Analysis: Titrate under nitrogen with 0.1 N methanolic potassium hydroxide VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N methanolic potassium hydroxide is equivalent to 9.61 mg of CH3SO3H. PROCEDURE 2 Solution A: 0.1 N citric acid solution, adjust with hydrochloric acid to a pH of 2.0 and mix.
USP 32
Diluent: Methanol and Solution A (1:1) Solution B: Acetonitrile and 0.01 M phosphate buffer, pH 7.0 (43:57) Solution C: Acetonitrile and 0.01 M phosphate buffer, pH 7.0 (3:2) Mobile phase: See the gradient table below.
Time (min) 0 18 30 40 41 Solution B (%) 100 100 0 0 100 Solution C (%) 0 0 100 100 0
Official Monographs / Bromocriptine 123

chloride CS, ferric chloride CS, cupric sulfate CS, and dilute hydrochloric acid (1 in 40): A: 3.0:3.0:2.4:31.6 B: 1.0:2.4:0.4:36.2 C: 0.6:2.4:0:37.0 Sample solution: 10 mg/mL of Bromocriptine Mesylate in methanol Procedure: Compare this solution with 10-mL portions of the Matching solutions in suitable matched tubes. Acceptance criteria: The solution is clear and not darker in color than Matching solutions A, B, and C. SPECIFIC ROTATION 781S: +95 to +105 Sample solution: 10 mg/mL, in a mixture of methylene chloride and methanol (1:1) LOSS ON DRYING : (See Thermal Analysis 891.) Determine the percentage of volatile substances by thermogravimetric analysis on an appropriately calibrated instrument, using 10 mg of Bromocriptine Mesylate. Heat the specimen under test at the rate of 10/min in an atmosphere of nitrogen at a flow rate of 45 mL/min. Record the thermogram from ambient temperature to 160: it loses NMT 4.0% of its weight ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, in a cold place. USP REFERENCE STANDARDS 11 USP Bromocriptine Mesylate RS
System suitability solution: 2.0 mg/mL of each of ergocryptine and Bromocriptine Mesylate in Diluent Standard solution: Dissolve USP Bromocriptine Mesylate RS in methanol, dilute quantitatively with an equal volume of Solution A, and dilute quantitatively and stepwise if necessary, with Diluent to obtain a solution having a concentration of 4.6 g per mL. Sample solution: 46 mg of Bromocriptine Mesylate, in a 10-mL volumetric flask, dissolve in 5.0 mL of methanol, dilute with Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 300 nm Column: 4.6-mm 15-cm; 3-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 15 between -ergocryptine and bromocriptine mesylate Tailing factor: NMT 1.5 Relative standard deviation: NMT 10.0% [NOTEThe Relative retention times are about 0.46 for -ergocryptine and 1.0 for bromocriptine mesylate.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Bromocriptine Mesylate taken: Result = (ri/rS) (CS/W) F 1000 = peak response for each impurity of the Sample solution rS = peak response for bromocriptine of the Standard solution CS = concentration of USP Bromocriptine Mesylate RS in the Standard solution (mg/mL) W = weight, in mg, of Bromocriptine Mesylate taken to prepare the Sample solution F = relative response factor is equal to 0.7 for any peaks eluting at a relative retention time of about 0.9 or less, and is equal to 1.0 for all other peaks Acceptance criteria Individual impurities: NMT 0.4% of bromcriptinin is found; NMT 0.1% of any individual impurity is found Total impurities: NMT 1.0% SPECIFIC TESTS COLOR OF SOLUTION 631 Matching solutions: Prepare three solutions, A, B, and C, containing, respectively, the following parts of cobaltous ri
Bromocriptine Mesylate Capsules

(Comment on this Monograph)id=m10234=Bromocriptine Mesylate Capsules=B-Monos.pdf) DEFINITION Bromocriptine Mesylate Capsules contain C32H40BrN5O5 CH4SO3 equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of C32H40BrN5O5. IDENTIFICATION The principal spot of the Sample solution corresponds, in RF value and color, to that from the Standard solution, as obtained under the Procedure for Organic Impurities. ASSAY PROCEDURE [NOTEConduct this procedure without exposure to daylight and with minimum exposure to artificial light.] Solution A: 0.125 mg/mL ammonium carbonate Mobile phase: Acetonitrile and Solution A (3:2) Standard solution: 1.0 mg/mL of bromocriptine from USP Bromocriptine Mesylate RS in dehydrated alcohol Sample solution: Remove, as completely as possible, the contents of NLT 10 Capsules. Weigh and determine the average weight/Capsule. Mix the combined contents, and transfer a weighed quantity of the powder, nominally equivalent to 50 mg of bromocriptine, to a 50-mL volumetric flask. Add 30 mL of dehydrated alcohol, and shake for 15 min. Dilute with dehydrated alcohol to volume, mix, and filter. [NOTEUse this solution without delay.] Chromatographic system (See Chromatography 621, System Suitability.)
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AU AS CS CU Mr1 Mr2
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= absorbance of the Sample solution = Absorbance of the Standard solution = concentration of USP Bromocriptine Mesylate RS in the Standard solution (mg/mL) = nominal concentration of bromocriptine in the Sample solution (mg/mL) = molecular weight of bromocriptine, 654.59 = molecular weight of bromocriptine mesylate, 750.70
Mode: LC Detector: UV 300 nm Column: 4-mm 25-cm; packing L7 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C32H40BrN5O5 in the Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bromocriptine Mesylate RS in the Standard solution (mg/mL) = nominal concentration of bromocriptine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 60 min Determine the amount of C32H40BrN5O5 CH4SO3 dissolved by the following: Sample solution: Sample per Dissolution 711, passed through a glass-fiber filter. Standard solution: USP Bromocriptine Mesylate RS at a known concentration in Medium. [NOTEA volume of alcohol not to exceed 5% of the total volume of the Standard solution may be used to bring the standard into solution before dilution with Medium.] Fluorometric conditions Excitation wavelength: 315 nm Emmision wavelength: 445 nm Blank: Medium Analysis Sample solutions: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C32H40BrN5O5 CH4SO3 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Procedure for content uniformity: [NOTEProtect all solutions from light.] Solution A: Dissolve 1.0 g of tartaric acid in 500 mL of water, add 500 mL of methanol, and mix. Standard solution: 0.04 mg/mL of USP Bromocriptine Mesylate RS in Solution A Sample solution: Transfer the contents of 1 Capsule into a 25-mL volumetric flask. Add 15 mL of Solution A, and shake by mechanical means for 20 min. Dilute with Solution A to volume, and mix. Filter and dilute 10.0 mL of the clear filtrate with Solution A to 50.0 mL. Spectrometric conditions Mode: UV Analytical wavelength: 306 nm Cell: 1 cm Blank: Solution A Analysis Samples: Blank, Sample solution, and Standard solution Calculate the percentage of C32H40BrN5O5 in the Capsule taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 rU rS CS
IMPURITIES Organic Impurities PROCEDURE [NOTEConduct this test without exposure to daylight and with minimum exposure to artificial light. Perform the test rapidly, preparing and spotting the Sample solution last.] Standard stock solution: 2.3 mg/mL of USP Bromocriptine Mesylate RS in methanol Standard solutions: Equivalent to 0.06, 0.04, 0.02, and 0.01 mg/mL of bromocriptine (equivalent to 3.0%, 2.0%, 1.0%, and 0.5%, respectively) from Standard stock solution diluted with methanol Sample solution: Equivalent to 20 mg of bromocriptine from Capsule contents, into a conical flask. Add 10 mL of methanol, and stir by mechanical means for 20 min. Centrifuge the suspension for 10 min at about 3500 rpm. The clear supernatant is the Sample solution. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: As 1.5-cm bands, 50-L portions of the Standard stock solution and of each of the four Standard solutions and 50 L of the Sample solution Developing solvent system: Methylene chloride, dioxane, alcohol, and ammonium hydroxide (180:15:5:1) Spray reagent: 2-in-1000 solution of o-phthalaldehyde in sulfuric acid Analysis Samples: Standard solution and Sample solution Develop under the exclusion of light in a tank lined with filter paper, previously equilibrated for 30 min, using Developing solvent system until the solvent front has moved a distance of 15 cm on the plate. Dry the plate briefly in a current of cold air. Spray evenly with the Spray reagent, and view the plate under long-wavelength UV light. Acceptance criteria: Any major secondary spot, other than the principal spot, obtained from the Sample solution is not greater in size and intensity than the spot obtained from the Standard solution corresponding to 3.0%, and any remaining spots are not greater in size and intensity than the spot obtained from the Standard solution corresponding to 1.0%. The sum of the related substances is NMT 5.0%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Bromocriptine Mesylate RS
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Sample solution: Sample per Dissolution711, passed through a glass-fiber filter. Standard solution: USP Bromocriptine Mesylate RS at a known concentration in Medium. [NOTEA volume of alcohol not to exceed 5% of the total volume of the Standard solution may be used to bring the standard into solution before dilution with Medium.] Fluorometric conditions Excitation wavelength: 315 nm Emmision wavelength: 445 nm Analysis Sample solutions: Standard solution and Sample solution Tolerances: NLT 80% (Q) of the labeled amount of C32H40BrN5O5 is dissolved. Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Medium: 0.1 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 30 min Mobile phase: Acetonitrile and 0.01 M ammonium carbonate (13:7) Sample solution: Sample per 711 Dissolution. Dilute with Medium to concentration that is similar to the Standard solution. Standard solution: USP Bromocriptine Mesylate RS in methanol, and quantitatively dilute with Dissolution Medium to obtain a solution having a known concentration similar to the expected concentration of the Sample solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 300 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C32H40BrN5O5 dissolved by comparison of the peak responses from the Standard solution and the solution under test. Tolerances: NLT 80% (Q) of the labeled amount of C32H40BrN5O5 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Procedure for content uniformity: [NOTEProtect all solutions from light.] Solution A: Dissolve 1.0 g of tartaric acid in 500 mL of water, add 500 mL of methanol, and mix. Standard solution: 0.04 mg/mL of USP Bromocriptine Mesylate RS in Solution A Sample solution: Transfer 1 Tablet into a 25-mL volumetric flask. Add 15 mL of Solution A, and shake by mechanical means for 30 min. Dilute with Solution A to volume, and mix. Filter and dilute 10.0 mL of the clear filtrate with Solution A to 50.0 mL.
Bromocriptine Mesylate Tablets

(Comment on this Monograph)id=m10260=Bromocriptine Mesylate Tablets=B-Monos.pdf) DEFINITION Bromocriptine Mesylate Tablets contain bromocriptine mesylate (C32H40BrN5O5 CH4SO3) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of bromocriptine (C32H40BrN5O5). IDENTIFICATION The principal spot from the Sample solution corresponds, in RF value and color, to that from the Standard solution, as obtained under the Procedure for Organic Impurities. ASSAY PROCEDURE [NOTEConduct this procedure without exposure to daylight and with minimum exposure to artificial light.] Mobile phase: Acetonitrile and 0.01 M ammonium carbonate (13:7) Standard solution: 0.22 mg/mL of USP Bromocriptine Mesylate RS in methanol Sample solution: Transfer an equivalent to 10 mg of bromocriptine, from powdered Tablets (NLT 20 Tablets), to an appropriate container. Add 40 mL of methanol, and stir for 20 min, protected from light. Quantitatively pass through a fine glass filtering funnel into a 50-mL volumetric flask. Rinse the filter with methanol, adding the rinsing to the filtrate, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 300 nm Column: 250- 4-mm stainless steel; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Coefficient of variation: NMT 3.0% for 3 replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C32H40BrN5O5 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bromocriptine Mesylate RS in the Standard solution (mg/mL) = nominal concentration of bromocriptine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 1. Medium: 0.1 N hydrochloric acid; 500 mL Apparatus 2: 120 rpm Time: 60 min Determine the amount of C32H40BrN5O5 CH4SO3 dissolved by: rU rS CS
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and any remaining spots are not greater in size and intensity than the spot obtained from the 1.0% Standard solution. The sum of the related compounds is NMT 5.0%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: The labeling indicates the Dissolution Test with which the product complies. USP REFERENCE STANDARDS 11 USP Bromocriptine Mesylate RS
Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 306 nm Cell: 1 cm Blank: Solution A Analysis Samples: Blank, Sample solution, and Standard solution Calculate the percentage of C32H40BrN5O5 in the Tablet taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 AU AS CS CU = absorbance of the solution from the Tablet = absorbance of the Standard solution = concentration of USP Bromocriptine Mesylate RS in the Standard solution (mg/mL) = nominal concentration of bromocriptine in the solution from the Tablet, based on the labeled quantity per Tablet and the extent of dilution (mg/mL) = molecular weight of bromocriptine, 654.59 = molecular weight of bromocriptine mesylate, 750.70
Bromodiphenhydramine Hydrochloride
(Comment on this Monograph)id=m10310=Bromodiphenhydramine Hydrochloride=B-Monos.pdf)
Mr1 Mr2
IMPURITIES Organic Impurities PROCEDURE [NOTEConduct this test without exposure to daylight and with minimum exposure to artificial light. Perform the test rapidly, preparing and spotting the Sample solution last.] Standard stock solution: 1.15 mg/mL of USP Bromocriptine Mesylate RS in methanol Standard solution: Equivalent to 0.1, 0.3, and 0.5 mg/mL of bromocriptine (equivalent to 1.0, 3.0, and 5.0%, respectively) from Standard stock solution in methanol Sample solution: Transfer an equivalent to 20 mg of bromocriptine, from powdered Tablets, to a conical flask. Add 10 mL of methanol, and stir by mechanical means for 20 min. Centrifuge the suspension for 10 min at about 4000 rpm. The clear supernatant is the Sample solution. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: As 1.5-cm bands, 10-L portions of the Standard stock solution and of each of the three Standard solutions and 50 L of the Sample solution Developing solvent system: Methylene chloride, dioxane, alcohol, and ammonium hydroxide (180:15:5:0.1) Spray reagent: 2-in-1000 solution of o-phthalaldehyde in sulfuric acid Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Dry the plate for 5 min in a current of cold air. Develop in a tank lined with filter paper, previously equilibrated for 20 min, using Developing solvent system until the solvent front has moved a distance of 10 cm on the plate. Dry the plate under vacuum at room temperature for 15 min. Spray evenly with the Spray reagent, and view the plate under long-wavelength UV light. Acceptance criteria: Any spot, other than the principal spot, from the Sample solution is not greater in size and intensity than the spot from the 3.0% Standard solution;
C17H20BrNO HCl 370.71 Ethanamine, 2-(4-bromophenyl)phenylmethoxy-N,N-dimethyl-, hydrochloride; 2-(p-Bromo--phenylbenzyl)oxy-N,N-dimethylethylamine hydrochloride [1808-12-4]. DEFINITION Bromodiphenhydramine Hydrochloride contains NLT 98.0% and NMT 101.0% of C17H20BrNO HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 228 nm Solution: 15 g/mL in 0.1 N sulfuric acid Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0%. ASSAY PROCEDURE Sample solution: Dissolve 700 mg of Bromodiphenhydramine Hydrochloride in 50 mL of glacial acetic acid, and add 10 mL of benzene and 15 mL of mercuric acetate TS. Add 2 drops of crystal violet TS. Analysis: Titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 37.07 mg of C17H20BrNO HCl. Acceptance criteria: 98.0%101.0% SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 148152 LOSS ON DRYING 731 Sample: Dry it at 105 for 3 h. Acceptance criteria: It loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Bromodiphenhydramine Hydrochloride RS
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Bromodiphenhydramine Hydrochloride Oral Solution

(Comment on this Monograph)id=m10330=Bromodiphenhydramine Hydrochloride Oral Solution=B-Monos.pdf) DEFINITION Bromodiphenhydramine Hydrochloride Oral Solution contains NLT 93.0% and NMT 107.0% of the labeled amount of bromodiphenhydramine hydrochloride (C17H20BrNO HCl). IDENTIFICATION INFRARED ABSORPTION 197K Sample solution: Transfer the final solution obtained from the titration in the Assay to a separator, add about 1 mL of 0.1 N sulfuric acid, and shake with 25 mL of ether. (Methyl red enters the ether phase.) Drain the aqueous layer into another separator, add 5 mL of 1 N sodium hydroxide, and shake with 10 mL of chloroform. Drain the chloroform layer into a small flask containing 2 g of anhydrous sodium sulfate, and swirl. Pour the chloroform solution through a small cotton pledget, pre-rinsed with chloroform, into a beaker, and evaporate to 5 mL. Apply a few drops of the solution directly to a potassium bromide plate, and completely remove the chloroform by warming for 23 min under an IR lamp. ASSAY PROCEDURE Sample solution: Evaporate a volume of Oral Solution nominally equivalent to 250 mg of bromodiphenhydramine hydrochloride to about half the original volume, using a suitable vacuum evaporator. Transfer the concentrated solution to a 250-mL separator, with the aid of sufficient warm water to bring the volume to the original volume. Add 20 g of sodium chloride, and shake until dissolved. Add 5 mL of 1 N sodium hydroxide, shake with 100 mL of ether, and drain the aqueous layer into a second separator containing 50 mL of ether. Shake, and discard the aqueous layer. Wash the ether solutions with two 20-mL portions of water, shaking each aqueous portion successively in the two separators, and then discard the aqueous solutions. Extract the ether solutions successively with 10.0 mL of 0.1 N sulfuric acid VS, followed by two 5-mL portions of water, and collect the aqueous extracts in a conical flask. Add methyl red TS to the solution in the flask. Analysis: Titrate the excess acid with 0.02 N sodium hydroxide VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.1 N sulfuric acid is equivalent to 37.07 mg of C17H20BrNO HCl. Acceptance criteria: 93.0%107.0% OTHER COMPONENTS ALCOHOL DETERMINATION, Method I 611: C2H5OH 12.0%15.0% of
Bromodiphenhydramine Hydrochloride and Codeine Phosphate Oral Solution

(Comment on this Monograph)id=m10340=Bromodiphenhydramine Hydrochloride and Codeine Phosphate Oral Solution=B-Monos.pdf) DEFINITION Bromodiphenhydramine Hydrochloride and Codeine Phosphate Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amounts of bromodiphenhydramine hydrochloride (C17H20 BrNO HCl) and codeine phosphate hemihydrate (C18H21NO3 H3PO4 1/2H2O). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 10 mg/mL of each of USP Bromodiphenhydramine Hydrochloride RS and USP Codeine Phosphate RS in methanol Sample solution: Transfer an equivalent to 10 mg of codeine phosphate, from a volume of Oral Solution, into a separator, and add 5 mL of water, 5 mL of methylene chloride, and 1 mL of ammonium hydroxide. Shake for 1 min, allow the layers to separate, and use the clear, lower layer. Developing solvent system: Alcohol and ammonium hydroxide (49:1) B. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol and water (4:1) Mobile phase: Methanol, water, 0.1 N ammonium hydroxide solution, and 0.1 N ammonium nitrate solution (27:3:2:1) Standard solution: 100 g/mL of USP Bromodiphenhydramine Hydrochloride RS and 80 g/mL of USP Codeine Phosphate RS in Diluent Sample solution: Nominally 10 mg/mL of bromodiphenhydramine hydrochloride and 0.08 mg/mL of codeine phosphate from Oral Solution diluted with Diluent. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30.0-cm; packing L3 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for bromodiphenhydramine and codeine are about 1.0 and 1.4, respectively.] Suitability requirements Resolution: NLT 2.0 between bromodiphenhydramine and codeine
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Bromodiphenhydramine Hydrochloride RS
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Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H20BrNO HCl in each mL of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response for bromodiphenhydramine from the Sample solution = peak response for bromodiphenhydramine from rS the Standard solution CS = concentration of USP Bromodiphenhydramine Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of the Sample solution (mg/mL) Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in each mL of Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response for codeine from the Sample solution = peak response for codeine from the Standard rS solution = concentration of USP Codeine Phosphate RS in CS the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight anhydrous codeine phosphate, Mr2 397.36 Acceptance criteria: 90.0%110.0% rU OTHER COMPONENTS ALCOHOL DETERMINATION, Method II 611: found. 4.0%6.0% is rU
Brompheniramine Maleate
(Comment on this Monograph)id=m10390=Brompheniramine Maleate=B-Monos.pdf)
C16H19BrN2 C4H4O4 435.31 2-Pyridinepropanamine, -(4-bromophenyl)-N,N-dimethyl-, ()-, (Z)-2-butenedioate (1:1); ()-2-p-Bromo--2-(dimethylamino)ethylbenzylpyridine maleate (1:1) [980-71-2]. DEFINITION Brompheniramine Maleate, dried at 105 for 3 h, contains NLT 98.0% and NMT 100.5% of C16H19BrN2 C4H4O4. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 35 g/mL in methanol ASSAY PROCEDURE Sample solution: 425 mg of Brompheniramine Maleate, previously dried, in 50 mL of glacial acetic acid. Add 1 drop of crystal violet TS. Analysis: Titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 21.77 mg of C16H19BrN2 C4H4O4. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Sample solution: 40 mg/mL of Brompheniramine Maleate in methylene chloride Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.2-m 4-mm; glass column containing 3% phase G3 on support S1AB Temperature: Column, 190; injection port and detector, 250
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Salmonella species, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. The total aerobic microbial count does not exceed 100 cfu/mL, and the total combined molds and yeasts count does not exceed 50 cfu/mL. PH 791: 4.56.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: Label it to indicate the alcohol content. USP REFERENCE STANDARDS 11 USP Bromodiphenhydramine Hydrochloride RS USP Codeine Phosphate RS
USP 32
Carrier gas: Dry helium Flow rate: Adjust to get a retention time of 67 min for the main peak. Injection size: 1 L System suitability Sample: Sample solution Suitability requirements Tailing factor: NMT 1.8 for the brompheniramine maleate peak Analysis Sample: Sample solution Record the chromatogram for a total time of NLT twice the retention time of the brompheniramine peak, and measure the areas of the peaks. Acceptance criteria: The total relative area of all extraneous peaks (except that of the solvent peak and maleic acid, if observed) is NMT 2.0%. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 130135 PH 791: 4.05.0, in a solution (1 in 100) LOSS ON DRYING 731: Drya sample at 105 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Brompheniramine Maleate RS CU
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= nominal concentration of brompheniramine maleate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 35.7 USP Endotoxin Units/mg of brompheniramine maleate PH 791: 6.37.3 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Brompheniramine Maleate RS USP Endotoxin RS
Brompheniramine Maleate Oral Solution

(Comment on this Monograph)id=m10400=Brompheniramine Maleate Oral Solution=B-Monos.pdf) DEFINITION Brompheniramine Maleate Oral Solution contains NLT 95.0% and NMT 105.0% of the labeled amount of brompheniramine maleate (C16H19BrN2 C4H4O4). IDENTIFICATION PROCEDURE Proceed as directed under IdentificationOrganic Nitrogenous Bases 181. Sample solution: Equivalent to 50 mg of brompheniramine maleate from a volume of Oral Solution, to a separator. Render distinctly alkaline with 1 N sodium hydroxide, and extract with two 50-mL portions of chloroform, shaking gently to avoid emulsification. Wash the combined chloroform extracts with 10 mL of water, and discard the aqueous phase. Filter the combined chloroform extracts into a conical flask, and evaporate the solvent on a steam bath, with the aid of a current of air. To the residue add 25 mL of dilute hydrochloric acid (1 in 1200), and proceed as directed under IdentificationOrganic Nitrogenous Bases 181, beginning with Transfer the liquid to a separator. Acceptance criteria: The Oral Solution meets the requirements. ASSAY PROCEDURE Sample solution: Equivalent to 20 mg of brompheniramine maleate from a volume of Oral Solution, to a separator. Render distinctly alkaline with 1 N sodium hydroxide, and extract with ten 10-mL portions of chloroform, shaking gently to avoid emulsification. Wash the combined chloroform extracts with 10 mL of water, wash the latter with 20 mL of chloroform, and discard the aqueous phase. Quantitatively filter the combined chloroform extracts and washings into a conical flask, and evaporate the solvent on a steam bath, with the aid of a current of air. To the residue add 25 mL of glacial acetic acid and 5 mL of acetic anhydride, agitate, and allow to stand for about 15 min. Add 1 drop of crystal violet TS. Analysis: Titrate with 0.01 N perchloric acid VS to a bluegreen endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.01 N perchloric acid is equivalent to 2.177 mg of brompheniramine maleate.
Brompheniramine Maleate Injection

(Comment on this Monograph)id=m10410=Brompheniramine Maleate Injection=B-Monos.pdf) DEFINITION Brompheniramine Maleate Injection is a sterile solution of Brompheniramine Maleate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C16H19BrN2 C4H4O4. IDENTIFICATION IDENTIFICATIONORGANIC NITROGENOUS BASES 181: Meets the requirements Sample solution: Equivalent to 50 mg of brompheniramine maleate from a volume of Injection, with dilute hydrochloric acid (1 in 1200) to 25 mL, and proceed as directed under IdentificationOrganic Nitrogenous Bases 181, beginning with Transfer the liquid to a separator. ASSAY PROCEDURE Analysis: Proceed with the Injection as directed under Salts of Organic Nitrogenous Bases 501 to prepare the solution used for the determination of the absorbance, AU, at 262 nm. For the determination of AS, dissolve 25 mg of USP Brompheniramine Maleate RS in 20 mL of dilute sulfuric acid (1 in 350), and treat this solution the same as the portion of Injection being assayed. Calculate the quantity, as a percentage, of C16H19BrN2 C4H4O4 in the Injection taken: Result = (AU/AS) (CS/CU) 100 AU AS CS = absorbance of Brompheniramine Maleate from the Sample solution = absorbance of USP Brompheniramine Maleate from the Standard solution = concentration of USP Brompheniramine Maleate RS in the Standard solution (mg/mL)
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95.0%105.0% 2.7%3.3% of Acceptance criteria: 95.0%105.0%
USP 32
OTHER COMPONENTS ALCOHOL DETERMINATION, Method I 611: C2H5OH SPECIFIC TESTS PH 791: 2.53.5
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Brompheniramine Maleate RS
Brompheniramine Maleate Tablets

(Comment on this Monograph)id=m10420=Brompheniramine Maleate Tablets=B-Monos.pdf) DEFINITION Brompheniramine Maleate Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of C16H19BrN2 C4H4O4. IDENTIFICATION Tablets meet the requirements under IdentificationOrganic Nitrogenous Bases 181. ASSAY PROCEDURE Sample solution: Weigh and finely powder NLT 20 Tablets. Weigh a portion of Tablets nominally equivalent to 4 mg of brompheniramine maleate, mix with 50 mL of water for 10 min, adjust with sodium hydroxide solution (1 in 10) to a pH of 11, and cool to room temperature. Extract the mixture with two 75-mL portions of solvent hexane, and combine the extracts in a second separator. Extract the solvent hexane solution with three 50-mL portions of dilute hydrochloric acid (1 in 120), combining the acid extracts in a 200-mL volumetric flask, and dilute with hydrochloric acid (1 in 120) to volume. Standard stock solution: 160 g/mL of USP Brompheniramine Maleate RS Standard solution: Transfer 25.0 mL of Standard stock solution to a separator containing 25 mL of water, mix, and proceed as directed under Sample solution, beginning with adjust with sodium hydroxide solution (1 in 10) to a pH of 11. The concentration of USP Brompheniramine Maleate RS in the Standard solution is 20 g/mL. Blank: Dilute hydrochloric acid (1 in 120) Spectrometric conditions Mode: UV Analytical wavelength: 264 nm Cell: 1 cm Analysis Samples: Blank, Sample solution, and Standard solution Calculate the percentage of C16H19Br N2 C4H4O4 in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance from the Sample solution = absorbance from the Standard solution = concentration of USP Brompheniramine Maleate RS in the Standard solution (g/mL) = nominal concentration of brompheniramine maleate in the Sample solution (g/mL)
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 500 mL Apparatus 1: 100 rpm Time: 45 min Standard solution: USP Brompheniramine Maleate RS at a known concentration in Medium Sample solution: Sample per 711 Dissolution, suitably diluted with 3 N hydrochloric acid. Spectrometric conditions Mode: UV Analytical wavelength: 264 Cuvette: 5 cm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C16H19BrN2 C4H4O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Brompheniramine Maleate RS
Brompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution

(Comment on this Monograph)id=m10427=Brompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution=BMonos.pdf) DEFINITION Brompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amounts of brompheniramine maleate (C16H19BrN2 C4H4O4) and pseudoephedrine sulfate [(C10H15NO)2 H2SO4]. IDENTIFICATION A. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sulfate 191 C. THIN-LAYER CHROMATOGRAPHY Standard solution: 1.2 mg/mL of USP Brompheniramine Maleate RS and 9 mg/mL of USP Pseudoephedrine Sulfate RS in methanol Sample solution: Equivalent to 6 mg of brompheniramine maleate in a separator. Add 0.5 mL of ammonium hydroxide and 5 mL of methylene chloride, shake for 1 min, and allow the layers to separate. Use the clear, lower layer as the Sample solution. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Ethyl ether, methanol, and ammonium hydroxide (16:3:1) Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the
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developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF values of the two principal spots from the Sample solution correspond to those from the Standard solution. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, tetrahydrofuran, and water (320:80:50:550) Transfer 1.0 mL of phosphoric acid, followed by 4.33 g of dodecyl sulfate sodium to this mixture, and mix. Adjust with ammonium hydroxide to a pH of 3.50 0.05 (see Chromatography 621, System Suitability). [NOTEThe pH of the Mobile phase is critical and may cause a 14 min difference in the retention times of the Internal standard solution and brompheniramine maleate.] Internal standard solution: 0.5 mg/mL of naphazoline hydrochloride in Mobile phase Solution P: 6000J g/mL of USP Brompheniramine Maleate RS in Mobile phase, J being the ratio of the labeled amount, in mg/mL, of brompheniramine maleate to the labeled amount, in mg/mL, of pseudoephedrine sulfate (Solution P) Standard solution: Transfer 30 mg of USP Pseudoephedrine Sulfate RS into a 25-mL volumetric flask. Add 5.0 mL each of Solution P and Internal standard solution, and dilute with Mobile phase to volume. [NOTEThe concentrations of the Standard solution are 1200J g of USP Brompheniramine Maleate RS/mL and about 1.2 mg of USP Pseudoephedrine Sulfate RS/mL.] Sample solution: Transfer a volume of Oral Solution, nominally equivalent to 30 mg of pseudoephedrine sulfate, to a 25-mL volumetric flask. Add 5.0 mL of Internal standard solution and dilute to volume with Mobile phase. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L11 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for pseudoephedrine sulfate, naphazoline hydrochloride, and brompheniramine maleate are 1.0, 1.5, and 2.5, respectively.] Suitability requirements Resolution: NLT 3 between the pseudoephedrine sulfate and naphazoline hydrochloride peaks; NLT 3 between the brompheniramine maleate and naphazoline hydrochloride peaks Relative standard deviation: NMT 2.0% Analysis Calculate the percentage of C16H19BrN2 C4H4O4 in the sample taken: Result = (RU/RS) CS/CU) 100 = peak response ratio for brompheniramine maleate to naphazoline hydrochloride from the Sample solution RS = peak response ratio for brompheniramine maleate to naphazoline hydrochloride from the Standard solution CS = concentration of USP Brompheniramine Maleate RS in the Standard solution (mg/mL) CU = nominal concentration of brompheniramine maleate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amounts of C16H19BrN2 C4H4O4 and [(C10H15NO)2 H2SO4] RU
Official Monographs / Budesonide 131

PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements for Oral Solution packaged in multiple-unit containers UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for Oral Solution packaged in multiple-unit containers ADDITIONAL REQUIREMENTS USP REFERENCE STANDARDS 11 USP Brompheniramine Maleate RS USP Pseudoephedrine Sulfate RS
Budesonide
(Comment on this Monograph)id=m10458=Budesonide=BMonos.pdf)
430.53 C25H34O6 Pregna-1,4-diene-3,20-dione, 16,17-butylidenebis(oxy)-11,21dihydroxy-, [11,16(R)], and 16,17-[(S)Butylidenebis(oxy)]-11,21-dihydroxypregna-1,4-diene-3,20dione; (RS)-11,16,17,21-Tetrahydroxypregna-1,4-diene-3,20-dione cyclic 16,17-acetal with butyraldehyde [51372-29-3; 51372-28-2; 51333-22-3]. DEFINITION Budesonide is a mixture of two epimeric forms, epimer A(C-22S) and epimer B(C-22R). It contains NLT 44.0% and NMT 51.0% of epimer A, and the sum of both epimers is NLT 98.0% and NMT 102.0% of C25H34O6, calculated on the dried basis. [NOTEProtect all solutions containing budesonide from light.] IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 25 g/mL Medium: Methanol ASSAY PROCEDURE Solution A: 3.17 mg/mL of monobasic sodium phosphate and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1. Mobile phase: Acetonitrile and Solution A (32:68) Standard solution: Dissolve a quantity of USP Budesonide RS in acetonitrile and dilute quantitatively with Solution A to obtain a solution having a concentration of 0.5 mg/mL, keeping the proportion of acetonitrile in this solution to NMT 30%. Sample solution: Dissolve 25 mg of Budesonide in 15 mL of acetonitrile in a 50-mL volumetric flask, and dilute to volume with Solution A. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention time for epimer A is 1.1 with respect to epimer B.]
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Analysis Sample: Sample solution Calculate the percentage of the 21-acetate of budesonide in the portion of C25H34O6 taken: Result = (rT1/rT2) 100 = sum of the peak areas for the two epimers of the 21-acetate of budesonide = sum of the areas of the two budesonide peaks rT2 Acceptance criteria: NMT 0.10% of the 21-acetate of budesonide is found. PROCEDURE 2: LIMIT OF 11-KETOBUDESONIDE Solution A: 3.17 mg/mL of monobasic sodium phosphate and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1. Mobile phase: Acetonitrile, isopropanol, and Solution A (26:9:65) Standard solution: Dissolve a quantity of USP Budesonide RS in acetonitrile and dilute quantitatively with Solution A to obtain a solution having a concentration of 0.5 mg/mL, keeping the proportion of acetonitrile in this solution to NMT 30%. Sample solution: Dissolve 25 mg of Budesonide in 15 mL of acetonitrile in a 50-mL volumetric flask, and dilute to volume with Solution A. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 3.5-m packing L1 Column temperature: 50 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for the two epimers of 11-ketobudesonide are 0.73 and 0.78, respectively; the relative retention times for 21-dehydrobudesonide, 14,15-dehydrobudesonide, and the first eluted epimer of budesonide (epimer B) are 0.68, 0.84, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.0 between the first epimer of 11ketobudesonide and 21-dehydrobudesonide; NLT 1.2 between the second epimer of 11-ketobudesonide and 14,15-dehydrobudesonide Column efficiency: NLT 5500 theoretical plates, determined from the budesonide epimer B peak Analysis Sample: Sample solution Calculate the percentage of the 11-ketobudesonide in the portion of C25H34O6 taken: rT1 Result = (rT1/rT2) 100 = sum of the peak areas for the two ketobudesonide peaks = sum of the areas of the two budesonide peaks rT2 Acceptance criteria: NMT 0.2% of the 11-ketobudesonide is found. PROCEDURE 3 Solution A: 3.17 mg/mL of monobasic sodium phosphate and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1 Mobile phase: Acetonitrile and Solution A (32:68) Standard solution: Dissolve a quantity of USP Budesonide RS in acetonitrile, and dilute quantitatively with Solution A to obtain a solution having a concentration of 0.5 mg/mL, rT1
Suitability requirements Resolution: NMT 1.5 between the two budesonide epimer peaks Column efficiency: NLT 5500 theoretical plates, determined from the budesonide epimer B peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of epimer A (C25H34O6) in the portion taken: Result = [rUA/(rUA + rUB)] 100 = epimer A peak area response from the Sample solution = epimer B peak area response from the Sample rUB solution Calculate the percentage of budesonide (C25H34O6) in the portion taken: Result = [(rUA + rUB)/(rSA + rSB)] (CS/CU) 100 = epimer A peak area response from the Sample solution = epimer B peak area response from the Sample rUB solution = epimer A peak area response from the Standard rSA solution = epimer B peak area response from the Standard rSB solution = concentration of USP Budesonide RS in the CS Standard solution (mg/mL) = concentration of Budesonide in the Sample CU solution (mg/mL) Acceptance criteria Epimer A: 44.0%51.0% Both epimers: 98.0%102.0% rUA IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF 21-ACETATE OF BUDESONIDE Solution A: 3.17 mg/mL of monobasic sodium phosphate and 0.23 mg/mL of phosphoric acid. The pH is 3.2 0.1. Mobile phase: Acetonitrile and Solution A (45:55) Standard solution: Dissolve a quantity of USP Budesonide RS in acetonitrile, and dilute quantitatively with Solution A to obtain a solution having a concentration of 0.5 mg/mL, keeping the proportion of acetonitrile in this solution to NMT 30%. Sample solution: Dissolve 25 mg of Budesonide in 15 mL of acetonitrile in a 50-mL volumetric flask, and dilute to volume with Solution A. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for the first eluted epimer of 21-acetate of budesonide, the second eluted epimer of 21-acetate of budesonide, the first eluted epimer of budesonide (epimer B), and the second eluted epimer of budesonide (epimer A) are 3.1, 3.2, 1.0, and 1.1, respectively.] Suitability requirements Column efficiency: NLT 5500 theoretical plates, determined from the budesonide epimer B peak rUA
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keeping the proportion of acetonitrile in this solution to NMT 30%. Sample solution: Dissolve 25 mg of Budesonide in 15 mL of acetonitrile in a 50-mL volumetric flask, and dilute to volume with Solution A. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 5500 theoretical plates, determined from the budesonide epimer B peak Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of C25H34O6 taken: Result = (rU/rT) 100 rU = peak area response for each impurity = sum of the responses of all of the peaks rT Acceptance criteria: The impurities meet the requirements listed in Impurity Table 1.
Impurity Table 1 Relative Retention Time 0.11 0.36 0.61; 0.66
d
Official Monographs / Bumetanide 133
Bumetanide
(Comment on this Monograph)id=m10463=Bumetanide=BMonos.pdf)
364.42 C17H20N2O5S Benzoic acid, 3-(aminosulfonyl)-5-(butylamino)-4-phenoxy-; 3-(Butylamino)-4-phenoxy-5-sulfamoylbenzoic acid [28395-03-1]. DEFINITION Bumetanide contains NLT 98.0% and NMT 102.0% of C17H20N2O5S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Sample solution: 50 g/mL in isopropyl alcohol C. The principal spot from the Sample solution exhibits an RF value corresponding to that of Standard solution A, as obtained in the Procedure for Organic Impurities. ASSAY PROCEDURE Sample solution: Dissolve 1 g of Bumetanide in 150 mL of alcohol, and add phenol red TS. Analysis: Titrate with 0.1 N sodium hydroxide VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N sodium hydroxide is equivalent to 36.44 mg of C17H20N2O5S. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1%, a 1-g specimen being used HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Standard solution A: 25 mg/mL of USP Bumetanide RS in methanol Standard solution B: 50 g/mL from Standard solution A in methanol Standard solution C: 50 g/mL of USP Bumetanide Related Compound B RS in methanol Standard solution D: 25 g/mL of USP Bumetanide Related Compound A RS in methanol Standard solution E: 25 g/mL of USP Butyl 3(butylamino)-4-phenoxy-5-sulfamoylbenzoate RS in methanol Sample solution: 25 mg/mL of Bumetanide in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Developing solvent system: Chloroform, cyclohexane, glacial acetic acid, and methanol (160:20:20:5)
Name 16-Hydroxylprednisolonea
D-Homobudesonideb
Relative Response Factor
Acceptance Criteria, NMT (%) 0.2 0.10 0.07 0.10 0.4 0.10 0.4
21-Dehydrobudesonide (epimers)c 14,15-Dehydrobudesonide Total specified impurities Any other individual impurity Total unspecified impurities
a
0.86
11,16,17,21-Tetrahydroxypregna-1,4-diene-3,20-dione. b 16,17-[(1RS)-Ethylidenebis(oxo)]-11,21-dihydroxypregna-1,4diene-3,20-dione. c 16,17-[(1RS)-Butylidenebis(oxo)]-11-hydroxy-3,20-dioxopregna-1,4dien-21-al. d 16,17-[(1RS)-Butylidenebis(oxo)]-11,21-dihydroxypregna-1,4,14triene-3,20-dione.
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62 Acceptance criteria Total aerobic microbial count: NMT 1000 cfu/g Total combined molds and yeast count: NMT 100 cfu/g LOSS ON DRYING 731 Analysis: Dry a sample at 105 to constant weight; it loses NMT 0.3% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Budesonide RS
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Internal standard stock solution: 0.5 mg/mL of 4ethylbenzaldehyde in methanol Internal standard solution: Add 10 mL of Internal standard stock solution, 10 mL of tetrahydrofuran, and 4.0 mL of glacial acetic acid to a 100-mL volumetric flask, and dilute to volume with methanol. Standard stock solution: 250 g/mL of USP Bumetanide RS in Internal standard solution Standard solution: 125 g/mL from Standard stock solution in water Sample solution: Transfer a volume equivalent to 250 g of bumetanide to a flask. Add an equal volume of Internal standard solution, and mix well. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times of ethylbenzaldehyde and bumetanide are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between the analyte and internal standard peaks Tailing factor: NMT 1.4 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H20N2O5S in each mL of the Injection: Result = (RU/RS) (CS/CU) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Bumetanide RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE Identification solution: 10 mg/mL of USP Bumetanide RS in methanol Standard solution A: 80 g/mL of USP Bumetanide RS from Identification solution, in methanol Standard solution B: 60 g/mL of USP Bumetanide RS from Standard solution A, in methanol Standard solution C: 40 g/mL of USP Bumetanide RS from Standard solution A, in methanol Standard solution D: 20 g/mL of USP Bumetanide RS from Standard solution A, in methanol Standard solution E: 10 g/mL of USP Bumetanide RS from Standard solution A, in methanol Standard solution F: 20 g/mL of USP Bumetanide Related Compound A RS, in methanol Sample solution: Transfer a volume of Injection, nominally equivalent to 5 mg of bumetanide, into a 125-mL separator, and adjust with 0.1 N sodium hydroxide to a pH of 12. Extract with two 20-mL portions of ethyl ether, discard the ethyl ether extracts, and adjust the aqueous layer with 1 N acetic acid to a pH of 4. Extract with two 20-mL portions of ethyl ether, passing the extracts through anhydrous sodium sulfate. Wash the sodium sulfate with about 5 mL of ethyl ether. Evaporate the combined ethyl RU RS CS
Visualization: Short-wavelength UV light Analysis Samples: Standard solution A, Standard solution B, Standard solution C, Standard solution D, Standard solution E, and Sample solution Proceed as directed in Chromatography 621, except after drying the application spots, place the plate in an unlined and unsaturated chromatographic chamber. Acceptance criteria: Any secondary spots from the Sample solution are not larger or more intense than the corresponding principal spots from the corresponding standard solution identified in Impurity Table 1. Individual impurities: See Impurity Table 1. Sum of other impurities: See Impurity Table 1.
Impurity Table 1 Corresponding Standard Solution Standard solution D Standard solution C Standard solution E Acceptance Criteria, NMT (%) 0.1 0.2 0.1
Name Bumetanide related compound A Bumetanide related compound B Paroxetine butyl 3(butylamino)-4-phenoxy-5sulfamoylbenzoate Other individual impurities Sum of other individual impurities
Standard solution B
0.2 0.4
SPECIFIC TESTS LOSS ON DRYING 731: Dry a sample at 105 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Bumetanide Related Compound A RS USP Bumetanide Related Compound B RS USP Bumetanide RS USP Butyl 3-(butylamino)-4-phenoxy-5-sulfamoylbenzoate RS
Bumetanide Injection
(Comment on this Monograph)id=m10465=Bumetanide Injection=B-Monos.pdf) DEFINITION Bumetanide Injection is a sterile solution of Bumetanide in Water for Injection, prepared with the aid of Sodium Hydroxide. It contains NLT 90.0% and NMT 110.0% of the labeled amount of bumetanide (C17H20N2O5S). IDENTIFICATION A. The relative retention time of the major peak in the Sample solution corresponds to that in the Standard solution, both relative to the internal standard, as obtained in the Assay. B. The principal spot from the Sample solution exhibits an RF value corresponding to that of the Identification solution, as obtained in the Procedure for Organic Impurities. ASSAY PROCEDURE Mobile phase: Methanol, tetrahydrofuran, glacial acetic acid, and water (50:5:2:45)
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ether extracts with the aid of a stream of nitrogen to dryness, and dissolve the residue in 0.5 mL of methanol. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 50 L Visualization: Short-wavelength UV light Developing solvent system: Chloroform, cyclohexane, glacial acetic acid, and methanol (160:20:20:5) Analysis Samples: Standard solution A, Standard solution B, Standard solution C, Standard solution D, Standard solution E, Standard solution F, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Acceptance criteria: Any secondary spots from the Sample solution are not larger or more intense than the corresponding principal spots from the corresponding standard solution identified in Impurity Table 1. Individual impurities: See Impurity Table 1. Sum of all other impurities: See Impurity Table 1.
Impurity Table 1 Corresponding Standard Solution Standard solution F Standard solutions AE Acceptance Criteria, NMT (%) 0.2 0.2 0.8
Official Monographs / Bumetanide 135

ASSAY PROCEDURE Mobile phase: Methanol, tetrahydrofuran, glacial acetic acid, and water (50:5:2:45) Internal standard stock solution: 0.5 mg/mL of 4ethylbenzaldehyde in methanol Internal standard solution: Add 10 mL of Internal standard stock solution, 10 mL of tetrahydrofuran, and 4.0 mL of glacial acetic acid to a 100-mL volumetric flask, and dilute with methanol to volume. Standard stock solution: 250 g/mL of USP Bumetanide RS in Internal standard solution Standard solution: 125 g/mL from Standard stock solution in water Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, nominally equivalent to 0.5 mg of bumetanide, to a 10-mL volumetric flask. Add 2.0 mL of Internal standard solution and sonicate for 5 min. Add 2.0 mL of water. Cool, and filter, discarding the first 1 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for 4ethylbenzaldehyde and bumetanide are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between the analyte and internal standard peaks Tailing factor: NMT 1.4 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H20N2O5S in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Bumetanide RS in the Standard solution (mg/mL) = nominal concentration of the bumetanide taken CU to prepare the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE Sample solution: Equivalent to 10 mg of bumetanide from powdered Tablets in a 50-mL centrifuge tube. Add 20 mL of acetone (spectrophotometric or HPLC quality), and shake by mechanical means for 10 min. Centrifuge for 10 min, decant the supernatant into a glass-stoppered, 25-mL conical flask, and evaporate with the aid of a stream of nitrogen to dryness. Dissolve the residue in 0.5 mL of methanol. Identification solution: 20 mg/mL of USP Bumetanide RS in methanol Standard solution A: 160 g/mL of USP Bumetanide RS from Identification solution, in methanol Standard solution B: 120 g/mL of USP Bumetanide RS from Standard solution A, in methanol Standard solution C: 80 g/mL of USP Bumetanide RS from Standard solution A, in methanol RU RS CS
Name Bumetanide related compound A Any other individual impurity Sum of all other individual impurities
SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 350 USP Endotoxin Units/mg of bumetanide PH 791: 6.87.8 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Bumetanide RS USP Bumetanide Related Compound A RS USP Endotoxin RS
Bumetanide Tablets
(Comment on this Monograph)id=m10468=Bumetanide Tablets=B-Monos.pdf) DEFINITION Bumetanide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of bumetanide (C17H20N2O5S). IDENTIFICATION A. The relative retention time of the major peak in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. B. The principal spot of the Sample solution exhibits an RF value corresponding to that of the Identification solution, as obtained in the Procedure for Organic impurities.
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Standard solution D: 40 g/mL of USP Bumetanide RS from Standard solution A, in methanol Standard solution E: 20 g/mL of USP Bumetanide RS from Standard solution A, in methanol Standard solution F: 0.04 mg/mL of USP Bumetanide Related Compound A RS in methanol Chromatographic system (see Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 25 L Visualization: Short-wavelength UV light Developing solvent system: Chloroform, cyclohexane, glacial acetic acid, and methanol (160:20:20:5) Analysis Samples: Sample solution, Standard solution A, Standard solution B, Standard solution C, Standard solution D, Standard solution E, and Standard solution F Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Acceptance criteria: Any secondary spots of the Sample solution are not larger or more intense than the corresponding principal spots of the corresponding Standard solution identified in the Impurity Table 1. Individual Impurities: See Impurity Table 1. Sum of all other impurities: See Impurity Table 1.
Impurity Table 1 Corresponding Standard Solution Standard solution F Standard solutions AE Acceptance Criteria, NMT (%) 0.2 0.2 0.8
Bupivacaine Hydrochloride
(Comment on this Monograph)id=m10490=Bupivacaine Hydrochloride=B-Monos.pdf)
C18H28N2O HCl H2O Change to read:
342.90
324.90 C18H28N2O HCl 2-Piperidinecarboxamide, 1-butyl-N-(2,6-dimethylphenyl)-, monohydrochloride, monohydrate, ()-; ()-1-Butyl-2,6-pipecoloxylidide monohydrochloride, monohydrate [v73360-54-0vUSP32]. Anhydrous [18010-40-7]. DEFINITION Bupivacaine Hydrochloride contains NLT 98.5% and NMT 101.5% of C18H28N2O HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197S Sample solution: Dissolve 230 mg in 15 mL of water in a separator, add 1 mL of 6 N ammonium hydroxide, and extract with three 30-mL portions of chloroform. Evaporate the chloroform at room temperature with the aid of a stream of nitrogen, and dry the residue in a vacuum. Add 2 mL of chloroform to the residue, and dissolve. B. ULTRAVIOLET ABSORPTION 197U Sample solution: 500 g/mL Medium: 0.1 N hydrochloric acid Analytical wavelength: 271 nm Acceptance criteria: Absorptivities do not differ by more than 3.0%, calculated on the anhydrous basis. C. IDENTIFICATION TESTSGENERAL, Chloride 191 Sample solution: Dissolve 50 mg in 10 mL of water in a small separator, render alkaline with 6 N ammonium hydroxide, and extract with 10 mL of ether. Acceptance criteria: The aqueous layer of the Sample solution meets the requirements. ASSAY PROCEDURE Sample: 600 mg Analysis: Transfer the Sample to a 250-mL conical flask, and dissolve in 20 mL of glacial acetic acid. Add 10 mL of mercuric acetate TS and 3 drops of crystal violet TS, and titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 32.49 mg of C18H28N2O HCl. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE 1: LIMIT OF RESIDUAL SOLVENTS Standard solution A: Pipet 2 mL of dehydrated alcohol into a 100-mL volumetric flask, dilute with water to volume, and mix. Transfer 2.0 mL of this solution to a 50-
Name Bumetanide related compound A Any other individual impurity Sum of all other individual impurities
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Stock solution A: 7.505 mg/mL of glycine and 5.85 mg/mL of sodium chloride in water Solution A: Stock solution A, 0.1 N hydrochloric acid, and water (4:1:45). Adjust, if necessary, with 0.1 N hydrochloric acid or 0.1 N sodium hydroxide to a pH of 2.9. Sample solutions: Sample per Dissolution 711. Dilute with Solution A as needed. Standard solution: USP Bumetanide RS at a known concentration in Medium Fluorometric conditions Excitation wavelength: 350 nm Emission wavelength: 450 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 85% (Q) of the labeled amount of C17H20N2O5S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Bumetanide RS USP Bumetanide Related Compound A RS
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mL volumetric flask, dilute with water to volume, and mix. The resulting solution contains 0.08% alcohol. Standard solution B: Pipet 2 mL of isopropyl alcohol into a 1000-mL volumetric flask, dilute with water to volume, and mix. Transfer 2.0 mL of this solution to a 100-mL volumetric flask, dilute with water to volume, and mix. The resulting solution contains 0.004% isopropyl alcohol. Sample solution: 40 mg/mL of Bupivacaine Hydrochloride Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 2-m; packing S3 Temperature Column: 175 Injection port: 200 Detector: 280 Flow rate: 40 mL/min Injection size: 5 L Carrier gas: Nitrogen Analysis Samples: Standard solution A, Standard solution B, and Sample solution Determine the percentage of alcohol taken: Result = 2(rU/rS) Determine the percentage of isopropyl alcohol taken: Result = 0.1(rU/rS) = peak response of the respective analytes in the Sample solution = peak response of the corresponding analytes in rS Standard solution A and Standard solution B Acceptance criteria: The sum of the content of alcohol and the content of isopropyl alcohol is NMT 2%. PROCEDURE 2 Solution A: Chloroform and isopropylamine (99:1) Sample solution: 20.0 mg/mL of Bupivacaine Hydrochloride in Solution A Standard solution A: 20.0 mg/mL of USP Bupivacaine Hydrochloride RS in Solution A Standard solution B: 100 g/mL of USP Bupivacaine Hydrochloride RS from Standard solution A diluted with Solution A Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Hexanes and isopropylamine (97:3) Samples: Sample solution, Standard solution A, and Standard solution B Analysis: Proceed as directed under Chromatography 621. Develop the chromatogram in a suitable chamber until the solvent has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and dry it in warm air. Place the plate in a closed chamber with a dish containing 1 g of iodine in a shallow layer, and allow to remain for about 5 min. Remove the plate from the chamber, spray it with 7 N sulfuric acid, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A, and the estimated size and intensity of any other spot of rU
Official Monographs / Bupivacaine 137

the Sample solution does not exceed that of the principal spot of the Standard solution B (0.5%). The total of the estimated sizes and intensities of all of the other spots of the Sample solution does not exceed four times that of the principal spot of the Standard solution B (2.0%). SPECIFIC TESTS PH 791 Sample solution: 1 in 100 Acceptance criteria: Between 4.5 and 6.0 WATER DETERMINATION, Method I 921: Between 4.0% and 6.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Bupivacaine Hydrochloride RS
Bupivacaine Hydrochloride Injection

(Comment on this Monograph)id=m10498=Bupivacaine Hydrochloride Injection=B-Monos.pdf) DEFINITION Bupivacaine Hydrochloride Injection is a sterile solution of Bupivacaine Hydrochloride in Water for Injection. It contains NLT 93.0% and NMT 107.0% of the labeled amount of C18H28N2O HCl. IDENTIFICATION A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Sample solution: Dilute a volume of Injection in 0.01 N hydrochloric acid to give a nominal concentration of 2 mg/mL of bupivacaine hydrochloride. Analysis: Proceed as directed in the General Chapter beginning with Transfer the liquid to a separator. Acceptance criteria: The Injection meets the requirements of the test. B. The retention time of the bupivacaine peak of the Sample solution corresponds to that of the bupivacaine peak of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.94 mg/mL of monobasic potassium phosphate and 2.48 mg/mL of dibasic potassium phosphate in water Adjust, if necessary, with 1 N potassium hydroxide or 1 M phosphoric acid to a pH of 6.8. Mobile phase: Acetonitrile and Solution A (65:35) Adjust, if necessary, with 1 M phosphoric acid to a pH of 7.7 0.2. Filter the solution through a membrane filter of 1-m or finer porosity and degas. Internal standard solution: 1.3 mg/mL of dibutyl phthalate in methanol Standard solution: Dissolve 50 mg of USP Bupivacaine Hydrochloride RS in 10.0 mL of water, using sonication if necessary, in a 100-mL volumetric flask. Add 10 mL of Internal standard solution, and dilute with methanol to volume. Sample solution: Transfer the amount of Injection equivalent to 50 mg of bupivacaine hydrochloride to a 100mL volumetric flask, add 10.0 mL of Internal standard solution, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.)
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solution of USP Dextrose RS in water, and (C) a solution of USP Bupivacaine Hydrochloride RS in Standard solution B to obtain solutions having concentrations corresponding to the labeled concentrations of bupivacaine hydrochloride and dextrose in the Injection. Sample solution: Bupivacaine Hydrochloride in Dextrose Injection Naphthalenediol reagent: Dissolve 20 mg of 1,3naphthalenediol in 10 mL of dehydrated alcohol containing 0.2 mL of sulfuric acid. Iodoplatinate reagent: Mix equal volumes of platinic chloride solution (3 in 1000) and potassium iodide solution (6 in 100). Application volume: See Analysis. Developing solvent system: Butyl alcohol, dehydrated alcohol, glacial acetic acid, and water (6:1:1:2) Analysis: Separately apply 10 L each of the Sample solution, Standard solution A, and Standard solution C to a portion of the chromatographic plate, and separately apply 1 L each of the Sample solution and Standard solution B to the remaining portion of the plate. Dry the applications in a current of warm air, develop the chromatograms in the Developing solvent system, remove the plate from the developing chamber, and mark the solvent front. Dry the plate in warm circulating air, and examine the plate under short-wavelength UV light. [NOTEAcceptance criteria 1 should be met.] Spray the plate with Naphthalenediol reagent, heat at 90 for 5 min, and examine the plate. [NOTEAcceptance criteria 2 should be met.] Cool the plate, spray it with Iodoplatinate reagent, and examine the plate. [NOTEAcceptance criteria 3 should be met. Also, bupivacaine appears as a blue-purple spot on a salmon-colored background, and the dextrose spots fade slightly.] Acceptance criteria 1: The RF value of the principal spot from the Sample solution corresponds to those of Standard solutions A and C. Acceptance criteria 2: The RF value of the principal bluepurple spot from the Sample solution corresponds to that from Standard solution B. Acceptance criteria 3: The RF value of the bupivacaine spot from the Sample solution corresponds to those from Standard solutions A and C. B. The retention time of the bupivacaine peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY BUPIVACAINE HYDROCHLORIDE Solution A: 1.94 mg/mL of monobasic potassium phosphate and 2.48 mg/mL of dibasic potassium phosphate Adjust, if necessary, with 1 N potassium hydroxide or 1 M phosphoric acid to a pH of 6.8. Mobile phase: Acetonitrile and Solution A (65:35) Adjust, if necessary, with 1 M phosphoric acid to a pH of 7.7 0.2. Pass the solution through a membrane filter of 1-m or finer porosity, and degas. Internal standard solution: 1.3 mg/mL of dibutyl phthalate in methanol Standard solution: Dissolve 50 mg of USP Bupivacaine Hydrochloride RS in 10.0 mL of water, using sonication if necessary, in a 100-mL volumetric flask. Add 10 mL of Internal standard solution and dilute with methanol to volume. Sample solution: Transfer the amount of Injection nominally equivalent to 50 mg of bupivacaine hydrochloride to a 100-mL volumetric flask, add 10.0 mL of Internal standard solution, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.)
Mode: LC Detector: UV 263 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for bupivacaine hydrochloride and dibutyl phthalate are about 1.0 and 1.2, respectively.] Suitability requirements Resolution: NLT 2.0 between bupivacaine hydrochloride and dibutyl phthalate Relative standard deviation: NMT 1.0% for the ratios of the bupivacaine hydrochloride peak to the dibutyl phthalate peak (three replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H28N2O HCl in the Injection: Result = (RU/RS) (CS/CU) 100 = peak response ratio of bupivacaine hydrochloride to the internal standard from the Sample solution = peak response ratio of bupivacaine hydrochloride RS to the internal standard from the Standard solution = concentration of USP Bupivacaine Hydrochloride CS RS, calculated on the anhydrous basis, in the Standard solution (mg/mL) = nominal concentration of bupivacaine CU hydrochloride taken to prepare the Sample solution (mg/mL) Acceptance criteria: 93.0%107.0% RU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 2.5 USP Endotoxin Units/mg of bupivacaine hydrochloride PH 791: 4.06.5 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass. Injection labeled to contain 0.5% or less of bupivacaine hydrochloride may be packaged in 50-mL, multiple-dose containers. USP REFERENCE STANDARDS 11 USP Bupivacaine Hydrochloride RS USP Endotoxin RS
Bupivacaine Hydrochloride in Dextrose Injection

(Comment on this Monograph)id=m10502=Bupivacaine Hydrochloride in Dextrose Injection=B-Monos.pdf) DEFINITION Bupivacaine Hydrochloride in Dextrose Injection is a sterile solution of Bupivacaine Hydrochloride and Dextrose in Water for Injection. It contains NLT 93.0% and NMT 107.0% of the labeled amounts of bupivacaine hydrochloride (C18H28N2O HCl) and dextrose (C6H12O6). It contains no preservative. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: Chromatographic silica gel mixture; 0.25 mm Standard solutions A, B, and C: Separately prepare (A) a solution of USP Bupivacaine Hydrochloride RS in water, (B) a
USP 32
Mode: LC Detector: UV 263 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for dibutyl phthalate and bupivacaine hydrochloride are about 1.2 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between bupivacaine hydrochloride and dibutyl phthalate Relative standard deviation: NMT 1.0% for the ratios of the bupivacaine hydrochloride peak to the dibutyl phthalate peak (three replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H28N2O HCl in the portion of Injection taken: Result = (RU/RS) (CS/CU) 100/L = peak response ratio of bupivacaine hydrochloride to the internal standard from the Sample solution RS = peak response ratio of bupivacaine hydrochloride to the internal standard from the Standard solution CS = concentration of USP Bupivacaine Hydrochloride RS, calculated on the anhydrous basis, in the Standard solution (mg/mL) CU = nominal concentration of bupivacaine hydrochloride in the Sample solution (mg/mL) L = label claim (mg/volume of Injection) Acceptance criteria: 93.0%107.0% DEXTROSE Analysis: Determine the angular rotation of the Injection in a suitable polarimeter tube (see Optical Rotation 781). Calculate the percentage of C6H12O6 in the portion of Injection taken: RU Result = {[A R)/F] 100} 100/L = 100 mm divided by the length of the polarimeter tube (mm) R = observed rotation (degrees) F = midpoint of the specific rotation range for anhydrous dextrose, 52.9 L = label claim of dextrose (g/100 mL) Acceptance criteria: 93.0%107.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 1.8 USP Endotoxin Units/mg of bupivacaine hydrochloride PH 791: Between 4.0 and 6.5 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Bupivacaine Hydrochloride RS USP Dextrose RS USP Endotoxin RS A
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Bupivacaine Hydrochloride and Epinephrine Injection

(Comment on this Monograph)id=m10505=Bupivacaine Hydrochloride and Epinephrine Injection=B-Monos.pdf) DEFINITION Bupivacaine Hydrochloride and Epinephrine Injection is a sterile solution of Bupivacaine Hydrochloride and Epinephrine or Epinephrine Bitartrate in Water for Injection. It contains NLT 93.0% and NMT 107.0% of the labeled amount of bupivacaine hydrochloride (C18H28N2O HCl). The content of epinephrine (C9H13NO3) does not exceed 0.001% (1 in 100,000). It contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amount of epinephrine (C9H13NO3). IDENTIFICATION A. PROCEDURE Sample solution 1: Dilute a volume of Injection in 0.01 N hydrochloric acid to give a concentration of 2 mg/mL of bupivacaine hydrochloride. Analysis 1: Proceed as directed under Identification Organic Nitrogenous Bases 181, beginning with Transfer the liquid to a separator. Acceptance criteria 1: The Injection meets the requirements of the test. Sample 2: Sample solution and Standard solution, as obtained from the Assay. Analysis 2: The retention time of the bupivacaine peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. PROCEDURE Sample: Volume of Injection, nominally equivalent to 50 g of epinephrine Analysis: Pipet the Sample into a suitable container, add 0.1 mL of Ferro-citrate solution and 2.0 mL of Buffer solution (prepared as directed under Epinephrine Assay 391), and allow the solution to stand for 10 min. Filter the solution. Acceptance criteria: The filtrate is violet in color and may turn brownish. ASSAY BUPIVACAINE HYDROCHLORIDE Solution A: 1.94 mg/mL of monobasic potassium phosphate and 2.48 mg/mL of dibasic potassium phosphate in water. Adjust, if necessary, with 1 N potassium hydroxide or 1 M phosphoric acid to a pH of 6.8. Mobile phase: Acetonitrile and Solution A (65:35). Adjust, if necessary, with 1 M phosphoric acid to a pH of 7.7 0.2. Filter the solution through a membrane filter of 1-m or finer porosity. Internal standard solution: 1.3 mg/mL of dibutyl phthalate in methanol Standard solution: Dissolve 50 mg of USP Bupivacaine Hydrochloride RS in 10.0 mL of water, using sonication if necessary, in a 100-mL volumetric flask. Add 10 mL of Internal standard solution, and dilute with methanol to volume. Sample solution: Transfer a volume of Injection, nominally equivalent to 50 mg of bupivacaine hydrochloride, to a 100mL volumetric flask. Add 10.0 mL of Internal standard solution, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.)
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Suitability requirements Resolution: NLT 6.0 between the epinephrine and dopamine peaks, System suitability solution Relative standard deviation: NMT 2.0% for replicate injections, Standard solution Analysis Samples: Standard solution and the Sample solution Calculate the percentage of C9H13NO3 in each mL of the Injection: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Epinephrine Bitartrate RS in the Standard solution (g/mL) = nominal concentration of epinephrine taken to CU prepare the Sample solution (g of epinephrine/mL) = molecular weight of epinephrine, 183.21 Mr1 = molecular weight of epinephrine bitartrate, Mr2 333.30 Acceptance criteria: 93.0%107.0% rU rS CS SPECIFIC TESTS COLOR AND CLARITY Standard solution: 0.1 N iodine VS and water (1:249) Sample: Bupivacaine Hydrochloride and Epinephrine Injection Analysis: Visually examine a portion of the Sample in a suitable clear glass test tube against a white background. [NOTEAcceptance criteria 1 should be met.] If any yellow color is observed in the Sample, concomitantly determine the absorbances of the Sample and the Standard solution in 1-cm cells with a suitable spectrophotometer set at 460 nm. [NOTEAcceptance criteria 2 should be met.] Acceptance criteria 1: The Sample is not pinkish, and it contains no precipitate. Acceptance criteria 2: The absorbance of the Sample does not exceed that of the Standard solution. BACTERIAL ENDOTOXINS TEST 85: NMT 1.6 USP Endotoxin Units/mg of bupivacaine hydrochloride PH 791: 3.35.5 INJECTIONS 1: It meets the requirements. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, protected from light. Injection labeled to contain 0.5% or less of bupivacaine hydrochloride may be packaged in 50-mL multiple-dose containers. LABELING: The label indicates that the Injection is not to be used if its color is pinkish or darker than slightly yellow or if it contains a precipitate. USP REFERENCE STANDARDS 11 USP Bupivacaine Hydrochloride RS USP Endotoxin RS USP Epinephrine Bitartrate RS
Mode: LC Detector: UV 263 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for bupivacaine hydrochloride and dibutyl phthalate are about 1.0 and 1.2, respectively.] Suitability requirements Resolution: NLT 2.0 between bupivacaine hydrochloride and dibutyl phthalate Relative standard deviation: NMT 1.0% for the ratios of the bupivacaine hydrochloride peak to the dibutyl phthalate peak (three replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H28N2O HCl in the Injection: Result = (RU/RS) (CS/CU) 100 = peak response ratio of bupivacaine hydrochloride to the internal standard peak from the Sample solution = peak response ratio of bupivacaine hydrochloride RS to the internal standard peak from the Standard solution = concentration of USP Bupivacaine Hydrochloride CS RS, calculated on the anhydrous basis, in the Standard solution (mg/mL) CU = nominal concentration of bupivacaine hydrochloride taken to prepare the Sample solution (mg/mL) Acceptance criteria: 93.0%107.0% EPINEPHRINE Mobile phase: Prepare a mixture of water, methanol, and 2 M monobasic sodium phosphate (900:50:50), containing 40 mg/mL of edetate disodium, 0.4 mL/L of phosphoric acid, and 0.4 mg/mL of sodium 1-octanesulfonate. Make adjustments, if necessary, to obtain a retention time of NLT 11 min for the epinephrine peak. System suitability solution: 2 g/mL of each of epinephrine bitartrate and dopamine hydrochloride in Mobile phase Standard solution: 2 g/mL of USP Epinephrine Bitartrate RS in Mobile phase Sample solution: Nominally equivalent to 1 g/mL of epinephrine, from a volume of Injection, diluted with Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Electrochemical detector held at a potential of +0.75 volt Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for epinephrine and dopamine are about 1.0 and 2.0, respectively, in the chromatograph of the System suitability solution. ] RU
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Column efficiency: NLT 6500 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Allow the Sample solution to elute for NLT two times the retention time of buprenorphine hydrochloride. Calculate the percentage of each impurity in the portion of Buprenorphine Hydrochloride taken: Result = (rU/rS) (CS/CU) 100
Buprenorphine Hydrochloride
(Comment on this Monograph)id=m10510=Buprenorphine Hydrochloride=B-Monos.pdf)
C29H41NO4 HCl 504.10 6,14-Ethenomorphinan-7-methanol, 17-(cyclopropyl-methyl)-(1,1-dimethylethyl)-4,5-epoxy-18,19-dihydro-3-hydroxy-6methoxy--methyl-, hydrochloride, [5,7 (S)]-; 21-Cyclopropyl-7-[(S)-1-hydroxy-1,2,2-trimethylpropyl]-6,14endo-ethano-6,7,8,14-tetrahydrooripavine hydrochloride [53152-21-9]. DEFINITION Buprenorphine Hydrochloride contains NLT 98.5% and NMT 101.0% of C29H41NO4 HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solution: 50 mg/mL of Buprenorphine Hydrochloride in methanol Analysis: To 0.5 mL of the Sample solution add 0.2 mL of a freshly prepared solution (1 in 10) of potassium ferricyanide TS and 0.5 mL of ferric chloride TS. Acceptance criteria: A blue color appears immediately. C. IDENTIFICATION TESTSGENERAL, Chloride 191 Sample solution: 1 in 100 ASSAY PROCEDURE Sample solution: 0.8 g of Buprenorphine Hydrochloride in 50 mL of glacial acetic acid. Add 10 mL of mercuric acetate. Analysis: Titrate Sample solution with 0.1 N perchloric acid VS, determining the green endpoint with 2 drops of crystal violet TS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 50.41 mg of C29H41NO4 HCl. Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Mobile phase: Methanol, 1% solution of ammonium acetate, and glacial acetic acid (60:10:0.01) Standard solution: 12.5 g/mL each of USP Buprenorphine Hydrochloride RS and USP Buprenorphine Related Compound A RS in Mobile phase Sample solution: 5 mg/mL of Buprenorphine Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 288 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 3.0 between buprenorphine hydrochloride and buprenorphine related compound A
= peak response for each impurity from the Sample solution rS = peak response of buprenorphine hydrochloride from the Standard solution CS = concentration of USP Buprenorphine Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of Buprenorphine Hydrochloride in the Sample solution (mg/mL) Acceptance criteria Individual impurity: NMT 0.25% Total impurities: NMT 0.65% rU SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 92 to 98 Sample solution: 20 mg/mL, in methanol PH 791: 4.06.0 in a solution containing 10 mg/mL WATER DETERMINATION, Method I 921: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Buprenorphine Hydrochloride RS USP Buprenorphine Related Compound A RS
Bupropion Hydrochloride
(Comment on this Monograph)id=m10520=Bupropion Hydrochloride=B-Monos.pdf)
C13H18ClNO HCl 276.21 1-Propanone, 1-(3-chlorophenyl)-2-[(1,1-dimethylethyl)amino]-, hydrochloride, ()-; ()-2-(tert-Butylamino)-3-chloropropiophenone hydrochloride [31677-93-7]. DEFINITION Bupropion Hydrochloride contains NLT 98.0% and NMT 102.0% of C13H18ClNO HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements for the silver nitrate precipate test
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1 mg/mL of Bupropion Hydrochloride
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Analysis Samples: Standard solutions and Sample solution Proceed as directed under the General Chapter. Locate and quantitate the spots obtained by scanning the entire plate with a suitable densitometer at 235 nm. Plot a standard curve of area versus concentrations of the Standard solutions. From the standard curve, determine the percentages of m-chlorobenzoic acid and any other impurity present. Acceptance criteria: NMT 0.2% of m-chlorobenzoic acid is found, and NMT 0.1% of any other individual impurity is found. PROCEDURE 2 Diluent, Solution A, Mobile phase, Solution B, Standard solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Bupropion Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response for each impurity from the Sample solution = peak response for bupropion hydrochloride from rS the Standard solution = concentration of USP Bupropion Hydrochloride CS RS in the Standard solution (mg/mL) = concentration of bupropion hydrochloride in the CU Sample solution (mg/mL) Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: The limits of impurities are specified in the accompanying table: NMT 0.3% of total unidentified impurities is found; and NMT 1.0% of total impurities is found, the results of Procedure 1 and Procedure 2 being added.
Impurity Table 1 Relative Retention Time Relative Response Factor 1.5 Acceptance Criteria, NMT (%) 0.5
Sample solution:
ASSAY PROCEDURE Diluent: Methanol and water (1:1) Solution A: Dissolve 6.8 g of monobasic potassium phosphate in 1900 mL of water. Adjust with 1 N sodium hydroxide to a pH of 7.0 and dilute with water to 2000 mL. Mobile phase: Methanol, tetrahydrofuran, and Solution A (39:11:51) Standard stock solution: 0.025 mg/mL of each of USP Bupropion Hydrochloride Related Compound A RS and USP Bupropion Hydrochloride Related Compound B RS in Diluent Standard solution: Transfer 25 mg of USP Bupropion Hydrochloride RS into a 25-mL volumetric flask. Dissolve in a portion of Diluent, pipet 2.0 mL of Solution B into the flask, and dilute with Diluent to volume. Sample solution: 1 mg/mL of Bupropion Hydrochloride in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 250 nm Column: 3.9-mm 15-cm; 5-m packing L7 Flow rate: 1.1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for bupropion hydrochloride related compound A, bupropion hydrochloride, and bupropion hydrochloride related compound B are about 0.92, 1.0, and 1.4, respectively.] Suitability requirements Resolution: NLT 1.3 between bupropion hydrochloride related compound A and bupropion hydrochloride Relative standard deviation: NMT 2.0% determined from bupropion hydrochloride; NMT 5.0% determined from bupropion hydrochloride related compound B Analysis Calculate the percentage of C13H18ClNO HCl in the portion of Bupropion Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Bupropion Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of bupropion hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Organic Impurities PROCEDURE 1 Standard solutions: 0.3, 0.2, and 0.1 mg/mL from Standard stock solution in methanol Sample solution: 100 mg/mL of Bupropion Hydrochloride in methanol Standard stock solution: 0.5 mg/mL of m-chlorobenzoic acid in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: A 0.25-mm layer of high-performance silica gel, previously washed with methanol Application volume: 4 L Developing solvent system: Toluene, cyclohexane, and glacial acetic acid (47:47:6) rU rS CS
rU
Name
2-(tert0.38 Butylamino)propiophenone hydrochloride 1-(3Chlorophenyl)-1,2propanedione 0.58
1.0
0.2
2-(tert0.71 Butylamino)-2chloropropiophenone hydrochloride 30.78 Chloropropiophenone Bupropion hydrochloride related compound A Bupropion
a
0.45
0.1
1.2 1.4
0.1 0.2
0.92
1.0
The percentage is determined by direct comparison to the area of the peak for bupropion hydrochloride related compound B obtained from the System suitability solution.
USP 32
Impurity Table 1 (continued) Relative Retention Time 1.14 Relative Response Factor Acceptance Criteria, NMT (%) 0.2a

concentration of nominally 3.0 mg/mL of bupropion hydrochloride. Add a portion of Diluent, equivalent to about one-half of the flask volume, and shake by mechanical means until the Tablets have disintegrated (between 30 and 60 min). Sonicate for 5 min, dilute with Diluent to volume, and mix. Allow to stand for at least 30 min, and pipet 10.0 mL of the supernatant into a 50-mL volumetric flask. Dilute with Diluent to volume, mix, and pass through a suitable filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 224 nm Column: 4.6-mm 15-cm; 5-m base-deactivated packing L1 Flow rate: 1.2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Calculate the pecentage of C13H18ClNO HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak area response from the Sample solution = peak area response from the Standard solution = concentration of USP Bupropion Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of bupropion hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Standard solution: USP Bupropion Hydrochloride RS in Medium at a known concentration Sample solution: Sample per Dissolution 711, diluted if necessary with 0.1 N hydrochloric acid. Spectrometric conditions Mode: UV Analytical wavelength: 252 nm Analysis Samples Standard solution and Sample solution Tolerances: NLT 80% (Q) of the labeled amount of C13H18ClNO HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Bupropion Hydrochloride RS rU rS CS
Name Bupropion hydrochloride related compound B
2-Bromo-31.63 chloropropiophenone 2-(tert2.30 Butylamino)-3,4chloropropiophenone hydrochloride 2-(tert2.74 Butylamino)-3,5chloropropiophenone hydrochloride Unknown

a
0.88 1.1
0.1 0.2
0.69
0.2
All other peaks
1.00
0.1, individual
The percentage is determined by direct comparison to the area of the peak for bupropion hydrochloride related compound B obtained from the System suitability solution.
NMT 0.5%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Bupropion Hydrochloride RS USP Bupropion Hydrochloride Related Compound A RS USP Bupropion Hydrochloride Related Compound B RS
Bupropion Hydrochloride Tablets

(Comment on this Monograph)id=m10523=Bupropion Hydrochloride Tablets=B-Monos.pdf) DEFINITION Bupropion Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of bupropion hydrochloride (C13H18ClNO HCl). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Crush 1 Tablet using a mortar and pestle. Prepare an approximate 1% (w/w) dispersion of the sample in potassium bromide. Acceptance criteria: The Sample shows strong bands at about 1690, 1560, and 1240 cm1 and a weaker band at about 740 cm1 similar to the reference preparation. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol and water (13:7) Buffer: 6.8 g of monobasic potassium phosphate and 1.164 g of sodium hydroxide in 1 L of water Mobile phase: Methanol and Buffer (13:7) Standard solution: 0.6 mg/mL of USP Bupropion Hydrochloride RS in Diluent Sample solution: Transfer a number of Tablets to a volumetric flask suitable to obtain a solution having a final
Bupropion Hydrochloride ExtendedRelease Tablets

(Comment on this Monograph)id=m10524=Bupropion Hydrochloride Extended-Release Tablets=B-Monos.pdf) DEFINITION Bupropion Hydrochloride Extended-Release Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of bupropion hydrochloride (C13H18ClNO HCl).
144
USP 32
Suitability requirements Resolution: NLT 1.5, between bupropion hydrochloride related compound C and bupropion hydrochloride related compound F, System Suitability solution A Tailing factor: NMT 1.9, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Relative response factor: Between 0.22 and 0.26 for mchlorobenzoic acid [NOTEUse the responses from System suitability solution B and Standard solution.] Analysis: Calculate the percentage of C13H18ClNO HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response for bupropion hydrochloride from the Sample solution = peak response for bupropion hydrochloride from rS the Standard solution = concentration of USP Bupropion Hydrochloride CS RS in the Standard solution (mg/mL) CU = nominal concentration of bupropion hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 For products labeled for dosing every 12 h Test 1 Medium: Water; 900 mL Sample solution: Sample per Dissolution 711, diluted with Medium as necessary. Apparatus 2: 50 rpm Times: 1, 4, and 8 h Standard solution: USP Bupropion Hydrochloride RS at a known concentration in the same Medium Spectrometric conditions Mode: UV spectrometry Analytical wavelength: 298 nm Cell: 1.0 cm Analysis Samples: Standard solution and Sample solution Tolerances: The percentage of the labeled amount of C13H18ClNO HCl dissolved at the times specified conforms to Acceptance Table 1.
Time (h) 1 4 8 Amount Dissolved 25%45% 60%85% NLT 80%
IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Crush 1 Tablet using a mortar and pestle. Prepare an approximate 1% (w/w) dispersion of the sample in potassium bromide. Acceptance criteria: The sample shows strong bands at about 1690, 1560, and 1240 cm1 and a weaker band at about 740 cm1 similar to the reference preparation. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol and 0.001 N hydrochloric acid (2:8) Solution A: Acetonitrile, trifluoroacetic acid, and water (10:0.04:90) Solution B: Acetonitrile, trifluoroacetic acid, and water (95:0.03:5) Mobile phase: See the gradient table below.
Time (min) 0 3.4 10.0 10.1 13.0 13.2 19.0 Solution A (%) 90 87 15 0 0 90 90 Solution B (%) 10 13 85 100 100 10 10
rU
System suitability solution A: Separate solutions of 0.20 mg/mL each of USP Bupropion Hydrochloride Related Compound C RS and USP Bupropion Hydrochloride Related Compound F RS in methanol. Transfer suitable volumes of each solution to a single volumetric flask to obtain 0.0018 mg/mL of USP Bupropion Hydrochloride Related Compound C RS and 0.018 mg/mL of USP Bupropion Hydrochloride Related Compound F RS. Dilute with Diluent to volume. System suitability stock solution B: 0.09 mg/mL of mchlorobenzoic acid in methanol System suitability solution B: 0.0018 mg/mL of mchlorobenzoic acid from System suitability stock solution B and Diluent Standard solution: 0.6 mg/mL of USP Bupropion Hydrochloride RS in Diluent Sample solution: Transfer a number of Tablets to a suitable homogenizer vessel containing sufficient methanol to obtain a concentration of 3.0 mg of bupropion hydrochloride/mL. Immediately homogenize the sample for 30 s at 20,000 rpm. Allow extraction for 3 min, and follow by two additional 10-s pulses, each at 20,000 rpm, pausing 3 min between these pulses to ensure complete extraction. Pass a portion of the solution through a nylon filter having a 0.45m porosity, discarding the first 24 mL of the filtrate. Pipet 10.0 mL of the filtrate into a 50-mL volumetric flask, and add about 25 mL of 0.001 N hydrochloric acid. Allow to cool to room temperature, and dilute with 0.001 N hydrochloric acid to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 226 nm Column: 4.6-mm 10-cm; 3.5-m packing L1 Flow rate: 1.5 mL/min Injection size: 5 L System suitability Samples: System suitability solution A, System suitability solution B, and Standard solution
Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Medium: 0.1 N hydrochloric acid, pH 1.5 (prepared by transferring 50 mL of concentrated hydrochloric acid to 6000 mL of water, adding 18 g of sodium hydroxide, mixing, and adjusting with either diluted sodium hydroxide or hydrochloric acid to a pH of 1.5 0.05); 900 mL Apparatus 1: 50 rpm Times: 1, 2, 4, and 6 h Determine the percentages of the labeled amount of C13H18ClNO HCl dissolved by using the following method. Buffer: 3.45 g of sodium phosphate monobasic monohydrate in 996 mL of water. Add 4.0 mL of triethylamine, and adjust with phosphoric acid to a pH of 2.80 0.05.
USP 32
Mobile phase: Methanol and Buffer (7:13) Standard solution: USP Bupropion Hydrochloride RS in Medium at a known concentration similar to the one expected in the Sample solution Sample solution: Use portions of the solution under test, and pass through a 0.45-m nylon filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 298 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Tolerances: The percentage of the labeled amount of C13H18ClNO HCl dissolved at the times specified conforms to Acceptance Table 2.
Time (h) 1 2 4 6 Amount Dissolved 25%50% 40%65% 65%90% NLT 80%

For products labeled for dosing every 24 h Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1: 75 rpm Times: 2, 4, 8, and 16 h Standard solution: USP Bupropion Hydrochloride RS at a known concentration in the same Medium Sample solution: Sample per Dissolution 711. Spectrometric conditions Mode: UV spectrometry Analytical wavelength: 252 nm Cell: 1.0 cm Analysis Samples: Standard solution and Sample solution Tolerances: The percentage of the labeled amount of C13H18ClNO HCl dissolved at the times specified conforms to Acceptance Table 4.
Time (h) 2 4 8 16 Amount Dissolved NMT 20% 20%45% 65%90% NLT 80%
Test 3: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 3. Medium, Apparatus, Standard solution, Sample solution, Spectometric conditions, and Analysis: Proceed as directed for Test 1, except the wavelength is about 250 nm. Times: 1, 2, 4, and 6 h Tolerances: The percentage of the labeled amount of C13H18ClNO HCl dissolved at the times specified conforms to Acceptance Table 3.
Test 6: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 6. Medium and Apparaus: Proceed as directed for Test 4. Times: 1, 2, 4, 8, and 12 h Standard solution: USP Bupropion Hydrochloride RS at a known concentration in Medium Spectrometric conditions Mode: UV spectrometry Analytical wavelength: 252 nm Cell: 1.0 cm Analysis: Determine the amount of C13H18ClNO HCl dissolved in filtered portions of the solution under test, suitably diluted with Medium, if necessary, in comparison with the Standard solution. Tolerances: The percentage of the labeled amount of C13H18ClNO HCl dissolved at the times specified conforms to Acceptance Table 6.
Time (h) 1 2 4 8 12 Amount Dissolved Between 15% and 35% Between 25% and 50% Between 40% and 65% Between 65% and 90% NLT 80%
Test 5: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 5. Medium and Analysis: Proceed as directed for Test 1. Spectrometric conditions: Proceed as directed for Test 1, except to use a 0.5-cm cell. Times: 1, 3, and 6 h Tolerances: The percentage of the labeled amount of C13H18ClNO HCl dissolved at the times specified conforms to Acceptance Table 5.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Procedure for content uniformity Standard solution: 0.33 mg/mL of USP Bupropion Hydrochloride RS in water Sample solution: Transfer 1 Tablet to a suitable homogenizer vessel containing a volume of water to obtain a concentration of about 0.33 mg of bupropion hydrochloride/mL. Immediately homogenize the sample using single 30-s pulses each at 5,000, 10,000, and 15,000 rpm, and follow by two pulses each at 20,000 rpm. After the homogenate has settled, mix at 5000 rpm for an additional 30 s. Pass a portion of the solution through a nylon filter having a 0.45-m porosity, discarding the first 4 mL of the filtrate.
146
USP 32
Analysis Sample solution:
Sample per Dissolution 711, filter
Buspirone Hydrochloride
(Comment on this Monograph)id=m10540=Buspirone Hydrochloride=B-Monos.pdf)
IMPURITIES Organic Impurities PROCEDURE Solution A, Solution B, Mobile phase, System suitability solution A, System suitability solution B, Standard solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay. Analysis: Calculate the percentage of each impurity in the portion of Tablets taken: Result = (rU/rS) (CS/CU) F 100 = peak response for each impurity from the Sample solution = peak response for bupropion hydrochloride from rS the Standard solution CS = concentration of USP Bupropion Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of bupropion hydrochloride in the Sample solution (mg/mL) F = relative response factor for each impurity (see Impurity Table 1 for values) Acceptance criteria: See Impurity Table 1. rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: When more than one Dissolution Test is given, the labeling states the Dissolution Test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Bupropion Hydrochloride RS USP Bupropion Hydrochloride Related Compound C RS USP Bupropion Hydrochloride Related Compound F RS
C21H31N5O2 HCl 421.96 8-Azaspiro[4,5]decane-7,9-dione, 8-[4-[4-(2-pyrimidinyl)-1piperazinyl]butyl]-, monohydrochloride; N-[4-[4-(2-Pyrimidinyl)-1-piperazinyl]butyl]-1,1cyclopentanediacetamide monohydrochloride [33386-08-2; 36505-84-7]. DEFINITION Buspirone Hydrochloride contains NLT 97.5% and NMT 102.5% of C21H31N5O2 HCl, calculated on the as-is basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The relative retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.36 mg/mL of monobasic potassium phosphate in water, adjust the solution with 10% sodium hydroxide (w/v) to a pH of 7.5, and filter
Impurity Table 1 Name 2-Amino-1-(3chlorophenyl)-1-propanone (3S,5S,6S)-6-(3Chlorophenyl)-6-hydroxy5-methyl-3-thiomorpholine carboxylic acid (3S,5R,6R)-6-(3Chlorophenyl)-6-hydroxy5-methyl-3-thiomorpholine carboxylic acid Bupropion Bupropion related compound F Bupropion related compound C m-Chlorobenzoic acid Bupropion related compound E Any unspecified impurity Total impurities Relative Retention Time 0.38 Relative Response Factor 0.80 Acceptance Criteria, NMT (%) 100 mg or less 0.3 150 mg or greater 0.3
0.56
0.86
1.0
1.5
0.78
0.88
0.5
0.4
1.0 1.71 1.75 1.80 2.25
0.55 0.59 0.24 1.00 1.00
1.2 0.3 0.3 0.4 0.2 3.2
2.3 0.3 0.3 0.4 0.2 3.3
USP 32
Mobile phase: Acetonitrile and Solution A (2:3) Internal standard stock solution: 2.5 mg/mL of propylparaben in methanol Internal standard solution: 0.125 mg/mL from Internal standard stock solution in water Standard stock solution: Dissolve 50 mg of USP Buspirone Hydrochloride RS in 25 mL of 1 N hydrochloric acid in a 100-mL volumetric flask; dilute with water to volume. Standard solution: 10.0 mL from Standard stock solution. Add 10.0 mL of Internal standard solution in a 50.0-mL volumetric flask, and add water to volume. Sample stock solution: Dissolve 50 mg of Buspirone Hydrochloride in 25 mL of 1 N hydrochloric acid in a 100mL volumetric flask; dilute with water to volume. Sample solution: 10.0 mL from Sample stock solution plus 10.0 mL of Internal standard solution in a 50.0-mL volumetric flask. Add water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for propylparaben and buspirone hydrochloride are about 0.55 and 1.0, respectively.] Suitability requirements Resolution: NLT 4 between buspirone hydrochloride and the internal standard Relative standard deviation: NMT 2.0% Analysis Calculate the percentage of C21H31N5O2 HCl in the portion of Buspirone Hydrochloride taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of buspirone hydrochloride to propylparaben from the Sample solution = peak response ratio of buspirone hydrochloride RS to propylparaben from the Standard solution = concentration of USP Buspirone Hydrochloride RS CS in the Standard solution (mg/mL) = concentration of buspirone hydrochloride in the CU Sample solution (mg/mL) Acceptance criteria: 97.5%102.5% RU OTHER COMPONENTS CONTENT OF CHLORIDE: Between 8.0% and 8.8% is found. Sample solution: 400 mg in 20 mL of water. Add 3 mL of nitric acid and 20.0 mL of 0.1 N silver nitrate VS. Gently boil the mixture for about 5 min. Filter, rinse the flask with about 80 mL of water divided into small portions, and filter each portion. Add 2 mL of 8% ferric ammonium sulfate. Analysis: While stirring rapidly, titrate the excess silver nitrate with 0.1 N ammonium thiocyanate VS to a faint redbrown endpoint. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.1 N silver nitrate consumed is equivalent to 3.545 mg of chloride.
Official Monographs / Buspirone 147

IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.5% HEAVY METALS, Method II 231: NMT 20 ppm SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Buspirone Hydrochloride RS
Buspirone Hydrochloride Tablets

(Comment on this Monograph)id=m10550=Buspirone Hydrochloride Tablets=B-Monos.pdf) DEFINITION Buspirone Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of buspirone hydrochloride (C21H31N5O2 HCl). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample solution: Grind 20 Tablets to a fine powder, add 50 mL of chloroform, stir for 35 min, and filter into a 250mL evaporating flask. Evaporate the solution with the aid of a rotary evaporator to dryness at low heat. Use the residue. B. The relative retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.36 mg/mL of monobasic potassium phosphate in water. Adjust the solution with 10% sodium hydroxide (w/v) to a pH of 7.5, and filter. Mobile phase: Acetonitrile and Solution A (2:3) Internal standard stock solution: 2.5 mg/mL of propylparaben in methanol Internal standard solution: 0.125 mg/mL from Internal standard stock solution in water Standard stock solution: Dissolve 50 mg of USP Buspirone Hydrochloride RS in 25 mL of 1 N hydrochloric acid in a 100-mL volumetric flask; dilute with water to volume. Standard solution: 10.0 mL from Standard stock solution. Add 10.0 mL of Internal standard solution in a 50.0-mL volumetric flask, and add water to volume. Sample stock solution: Equivalent to 100 mg of buspirone hydrochloride from powdered Tablets, in a 200-mL volumetric flask. Add 50 mL of 1 N hydrochloric acid, and shake for 15 min. Add about 100 mL of water, and shake for 30 min. Dilute with water to volume, mix, and filter, discarding the first 20 mL of the filtrate.
148
Buspirone / Official Monographs
USP 32
Sample solution: Transfer 10.0 mL of the Sample stock solution and 10.0 mL of Internal standard solution into a 50mL volumetric flask. Dilute with water to volume, and mix. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for propylparaben and buspirone hydrochloride are 0.55 and 1.0, respectively.] Suitability requirements Resolution: NLT 4 between buspirone hydrochloride and the internal standard Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H31N5O2 HCl in the Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of buspirone hydrochloride to propylparaben from the Sample solution = peak response ratio of buspirone hydrochloride RS to propylparaben from the Standard solution = concentration of USP Buspirone Hydrochloride CS RS in the Standard solution (mg/mL) = nominal concentration of buspirone CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 30 min Sample solutions: Sample per Dissolution 711. Dilute with Medium as needed at a known concentration. Standard solution: USP Buspirone Hydrochloride RS in Medium Spectometric conditions Mode: UV Analytical wavelength: 235 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 80% (Q) of the labeled amount of C21H31N5O2.HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Buspirone Hydrochloride RS
Busulfan
(Comment on this Monograph)id=m10570=Busulfan=BMonos.pdf)
C6H14O6S2 1,4-Butanediol, dimethanesulfonate; Butane-1,4-diyl dimethanesulfonate [55-98-1].
246.30
DEFINITION Busulfan contains NLT 98.0% and NMT 100.5% of C6H14O6S2, calculated on the dried basis. IDENTIFICATION A. PROCEDURE Sample: 100 mg Analysis: Fuse Sample with about 100 mg of potassium nitrate and a pellet of potassium hydroxide weighing approximately 250 mg. Cool, dissolve the residue in water, acidify with 3 N hydrochloric acid, and add a few drops of barium chloride TS. Acceptance criteria: A white precipitate is formed. B. PROCEDURE Sample: 100 mg Analysis: Add 10 mL of water and 5 mL of 1 N sodium hydroxide. Heat until a clear solution is obtained. Acceptance criteria: An odor characteristic of methanesulfonic acid is perceptible. C. PROCEDURE Sample solution: Use the solution from Identification B. Analysis: Cool the Sample solution obtained in Identification test B, and divide it into two equal portions. To one portion add 1 drop of potassium permanganate TS. Acidify the second portion of the solution with 2 N sulfuric acid, and add 1 drop of potassium permanganate TS. Acceptance criteria: The first portion becomes purple which changes to violet, then to blue and finally to emerald green. In the second portion (acidified) the color of the permanganate is not discharged. ASSAY PROCEDURE Sample solution: 80 mg of Busulfan in a 250-mL conical flask. Add about 30 mL of water, swirl, add phenolphthalein TS, and neutralize with 0.05 N sodium hydroxide. Connect the flask to a reflux air condenser, and boil the mixture gently for NLT 30 min, adding water occasionally to maintain the volume. Cool to room temperature, and add phenolphthalein TS.
USP 32
Analysis: Titrate with 0.05 N sodium hydroxide VS. Each mL of 0.05 N sodium hydroxide is equivalent to 6.158 mg of C6H14O6S2. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
Official Monographs / Butabarbital 149

Transfer an equivalent to 80 mg of busulfan, from powdered Tablets (NLT 40), to a 100-mL beaker. Extract with four 20mL portions of acetone, each time stirring the mixture well, then allowing the insoluble matter to settle, and finally decanting the supernatant through a sintered-glass filter into a 250-mL conical flask. Evaporate the combined acetone extracts to about 10 mL, add phenolphthalein TS, and neutralize with 0.05 N sodium hydroxide. Evaporate to dryness, and add about 30 mL of water. Connect the flask to a reflux air condenser, and boil the mixture gently for NLT 30 min, adding water occasionally to maintain the volume. Cool to room temperature, and add phenolphthalein TS. Analysis: Titrate with 0.05 N sodium hydroxide VS. Each mL of 0.05 N sodium hydroxide is equivalent to 6.158 mg of C6H14O6S2. Acceptance criteria: 93.0%107.0% PERFORMANCE TESTS DISINTEGRATION 701: 30 min, the use of disks being omitted UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
NMT 0.1%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 115118 LOSS ON DRYING 731: Dry it in a vacuum at 60 to constant weight: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label bears a warning that great care should be taken to prevent inhaling particles of Busulfan and exposing the skin to it.
Busulfan Tablets
(Comment on this Monograph)id=m10600=Busulfan Tablets=BMonos.pdf) DEFINITION Busulfan Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of C6H14O6S2. IDENTIFICATION A. PROCEDURE Sample: A suitable number of Tablets Analysis: Pulverize the Sample and extract the powder with several portions of acetone. Evaporate the combined acetone extracts, with the aid of a current of air, on a steam bath. Acceptance criteria: The dry residue melts at about 115. B. PROCEDURE Sample: 100 mg of the powder obtained in Identification A Analysis: Fuse the Sample with 100 mg of potassium nitrate and a pellet of potassium hydroxide weighing 250 mg. Cool, dissolve the residue in water, acidify with 3 N hydrochloric acid, and add a few drops of barium chloride TS. Acceptance criteria: A white precipitate is formed. C. PROCEDURE Sample: 100 mg of the powder obtained in Identification A Analysis: To the Sample add 10 mL of water and 5 mL of 1 N sodium hydroxide. Heat until a clear solution is obtained. Acceptance criteria: An odor characteristic of methanesulfonic acid is perceptible. D. PROCEDURE Sample solution: Use the solution from Identification C. Analysis: Cool the Sample solution, and divide it into two equal portions. To one portion add 1 drop of potassium permanganate TS. Acidify the second portion of the Sample solution with 2 N sulfuric acid, and add 1 drop of potassium permanganate TS. Acceptance criteria: The first portion becomes purple which changes to violet, then to blue, and finally to emerald-green. In the second portion (acidified), the color of the permanganate is not discharged. ASSAY PROCEDURE Sample solution: [NOTEGuard against accidental inhalation of the fine powder.]
Butabarbital
(Comment on this Monograph)id=m10640=Butabarbital=BMonos.pdf)
212.25 C10H16N2O3 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-5-(1-methylpropyl)-; 5-sec-Butyl-5-ethylbarbituric acid [125-40-6]. DEFINITION Butabarbital contains NLT 98.5% and NMT 101.0% of C10H16N2O3, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197M ASSAY PROCEDURE Internal standard solution: 2 mg/mL of tetracosane in chloroform Standard stock solution: 2 mg/mL of USP Butabarbital RS in chloroform Standard solution: 1 mg/mL from Standard stock solution diluted with Internal standard solution Sample stock solution: 2 mg/mL of Butabarbital in chloroform Sample solution: 1 mg/mL from Sample stock solution diluted with Internal standard solution Chromatographic system (See Chromatography 621, System Suitability.)
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Standard solution B, corresponding to NMT a total of 1% of impurities. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class Ia 741: 164167 LOSS ON DRYING 731: Dry a sample at 105 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Butabarbital RS
Mode: GC Detector: Flame ionization Column: 4-mm 1.8-m Packing: 10% phase G37 on support S1AB Temperature Column: 260 Injection port: 260 Detector block: 300 Carrier gas: Dry nitrogen Flow rate: 50 mL/min Injection size: 2 L System suitability Sample: Standard solution [NOTEThe relative retention times for butabarbital and tetracosane are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the Butabarbital and internal standard peaks Tailing factor: NMT 1.3 for Butabarbital and NMT 1.2 internal standard peaks Relative standard deviation: NMT 1.0% Analysis Calculate the percentage of C10H16N2O3 in the portion of Butabarbital taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Butabarbital RS in the Standard solution (mg/mL) = concentration of Butabarbital in the Sample CU solution (mg/mL) Acceptance criteria: 98.5%101.0% RU RS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Standard solution A: 4.0 mg/mL of USP Butabarbital RS in a mixture of chloroform and methanol (1:1) Standard solution B: 0.4 mg/mL from Standard solution A in a mixture of chloroform and methanol (1:1) Sample solution: 40 mg/mL of Butabarbital in a mixture of chloroform and methanol (1:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Acetone, methylene chloride, methanol, and ammonium hydroxide (5:3:1:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed under the General Chapter. Develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, and dry in a current of air. Spray the plate with a solution of mercurous nitrate dihydrate in 0.15 N nitric acid (1 in 100), and immediately estimate the intensities of any spots in the chromatogram of the Sample solution, other than the principal spot, in comparison with Standard solution B. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A; and the sum of the intensities of any secondary spots observed in the chromatogram of the Sample solution is NMT the intensity of the principal spot produced by
Butabarbital Sodium
(Comment on this Monograph)id=m10650=Butabarbital Sodium=B-Monos.pdf) 234.23 C10H15N2NaO3 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-ethyl-5-(1-methylpropyl)-, monosodium salt; Sodium 5-sec-butyl-5-ethylbarbiturate [143-81-7]. DEFINITION Butabarbital Sodium contains NLT 98.2% and NMT 100.5% of C10H15N2NaO3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K: 150 mg to a suitable separator, dissolve in 10 mL of water, and add 15 mL of 3 N hydrochloric acid. Extract with three 20-mL portions of chloroform, filter the extracts through anhydrous sodium sulfate, and collect the extracts in a suitable beaker. Evaporate the combined chloroform extracts on a steam bath with the aid of a current of air to dryness, and dry the residue at 105 for 2 h. B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 240 nm Standard solution: 10 g/mL in pH 9.6 alkaline borate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions) Sample solution: 10 g/mL in pH 9.6 alkaline borate buffer Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0% C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements Analysis: Ignite about 100 mg. ASSAY PROCEDURE Standard stock solution: 0.125 mg/mL of USP Butabarbital RS in pH 9.6 Alkaline Borate Buffer (see Reagents, Indicators, and SolutionsBuffer Solutions) Standard solution: 0.0125 mg/mL from Standard stock solution in pH 9.6 Alkaline Borate Buffer Sample stock solution: 0.14 mg/mL of Butabarbital Sodium in pH 9.6 Alkaline Borate Buffer Standard solution: 0.014 mg/mL from Sample stock solution in pH 9.6 Alkaline Borate Buffer Spectrometric conditions Mode: UV Analytical wavelength: 240 nm Samples: Standard solution and Sample solution Analysis: Calculate the percentage of C10H15N2NaO3 in the portion of Butabarbital Sodium taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 AS AU = absorbance of the solution from the Standard solution = absorbance of the solution from the Sample solution
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= concentration of USP Butabarbital RS in the Standard solution (g/mL) = concentration of Butabarbital Sodium in the CU Sample solution (g/mL) = molecular weight of butabarbital sodium, 234.23 Mr1 = molecular weight of butabarbital, 212.25 Mr2 Acceptance criteria: 98.2%100.5% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 30 ppm Organic Impurities PROCEDURE Standard solution A: 4.0 mg/mL of USP Butabarbital RS in a mixture of chloroform and methanol (1:1) Standard solution B: 0.4 mg/mL from Standard solution A in a mixture of chloroform and methanol (1:1) Sample solution: 44 mg/mL of Butabarbital Sodium in a mixture of chloroform and methanol (1:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Acetone, methylene chloride, methanol, and ammonium hydroxide (5:3:1:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed under the General Chapter. Develop the chromatogram in a solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, and in a current of air. Spray the plate with a solution of mercurous nitrate dihydrate in 0.15 N nitric acid (1 in 100), and immediately estimate the intensities of any spots in the chromatogram of the Sample solution, other than the principal spot, in comparison with Standard solution B. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A; and the sum of the intensities of any secondary spots observed in the chromatogram of the Sample solution is NMT the intensity of the principal spot produced by Standard solution B, corresponding to NMT a total of 1% of impurities. SPECIFIC TESTS COMPLETENESS OF SOLUTION: 1.0 g in 10 mL of carbon dioxide-free water: after 1 min, the solution is clear and free from undissolved solid PH 791: 10.011.2, in the solution prepared for the test for Completeness of Solution LOSS ON DRYING 731: Dry a sample at 150 to constant weight: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Butabarbital RS CS
Official Monographs / Butabarbital 151

IDENTIFICATION INFRARED ABSORPTION 197K Sample: Equivalent to 150 mg of butabarbital sodium from a volume of Oral Solution, to a separator. Render it distinctly alkaline by the addition of 1 N sodium hydroxide, and saturate it with sodium chloride. Extract the mixture with two 15-mL portions of ether, and discard the ether. Acidify the solution with hydrochloric acid, and render it just alkaline to litmus by adding small portions of sodium bicarbonate (carbonate-free). Extract the liberated acid barbiturate using five 20-mL portions of chloroform. Wash the combined chloroform extracts with 10 mL of water acidified with 1 drop of hydrochloric acid, then extract the water with 10 mL of chloroform, adding the latter to the main chloroform solution. Pass the chloroform solution through a pledget of cotton or other suitable filter, previously washed with chloroform, into a tared beaker, and finally wash the separator and the filter with three 5-mL portions of chloroform. Evaporate the combined chloroform solution and washings on a steam bath with the aid of a current of air to dryness, and dry the residue at 105 for 2 h. ASSAY PROCEDURE Internal standard solution: 0.7 mg/mL of secobarbital in chloroform Standard solution: 1 mg/mL of USP Butabarbital RS and 1.4 mg/mL of secobarbital in chloroform Sample solution: Equivalent to 30 mg of butabarbital sodium from a volume of Oral Solution, to a separator. Add 1 mL of bromine water (prepared by dissolving 2.0 mL of bromine and 10 g of potassium bromide in 60 mL of water), and swirl. Allow to stand for 5 min, add 1 mL of sodium metabisulfite solution (1 in 10), and swirl. Add 300 mg of sodium bicarbonate in small portions, with mixing, and extract with four 10-mL portions of chloroform. Filter the extracts through about 15 g of anhydrous sodium sulfate that is supported on a funnel by a small pledget of glass wool. Collect the combined filtrates in a 50-mL volumetric flask, wash the sodium sulfate with 5 mL of chloroform, collecting the washing with the filtrate, dilute with chloroform to volume, and mix. Combine 2.0 mL of this solution with 2.0 mL of Internal standard solution in a suitable container, and reduce the volume to about 1 mL by evaporation, with the aid of a stream of dry nitrogen, at room temperature. [NOTEThis solution includes a bromination step for elimination of parabens and a carbonatechloroform extraction for elimination of benzoic acid.] Chromatographic system and System suitability: Proceed as directed for Chromatographic System and System Suitability under Barbiturate Assay 361. Resolution: NLT 2.4 between butabarbital and secobarbital [NOTEThe relative retention times for butabarbital and secobarbital are 0.6 and 1.0, respectively.] Analysis Samples: Standard solution and Sample solution Proceed as directed for Barbiturate Assay 361, Analysis. Calculate the percentage of C10H15N2NaO3 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Butabarbital RS in the Standard solution (g/mL) = nominal concentration of butabarbital sodium in the Sample solution (g/mL) = molecular weight of butabarbital sodium, 234.23
Butabarbital Sodium Oral Solution

(Comment on this Monograph)id=m10670=Butabarbital Sodium Oral Solution=B-Monos.pdf) DEFINITION Butabarbital Sodium Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of butabarbital sodium (C10H15N2NaO3).
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Analysis Samples: Standard solution and Sample solution Proceed as directed for Barbiturate Assay 361, Procedure. Calculate the percentage of C10H15N2NaO3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Butabarbital RS in the Standard solution (mg/mL) = nominal concentration of butabarbital sodium in CU the Sample solution (mg/mL) Mr1 = molecular weight of butabarbital sodium, 234.23 = molecular weight of butabarbital, 212.25 Mr2 Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Sample solutions: Sample per Dissolution 711. Mix with sufficient ammonium hydroxide to provide a concentration of 0.5 N ammonium hydroxide. Dilute with Medium as necessary. Standard solution: USP Butabarbital RS in Medium to a known concentration Spectrometric conditions Mode: UV Analytical wavelength: 239 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C10H15N2NaO3 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Spectrometric conditions Detector: UV Analytical wavelength: 240 nm Solution A: Methanol and 1 N hydrochloric acid (9:1) Standard solution: 0.45 mg/mL of USP Butabarbital RS in Solution A Sample solution: Transfer 1 finely powdered Tablet to a 25mL volumetric flask. Add Solution A to volume, and mix. Filter, discarding the first 5 mL of the filtrate, and dilute the subsequent filtrate, if necessary, with Solution A to obtain a solution containing 0.50.6 mg of butabarbital sodium/mL. Analysis: Transfer 2.0 mL each of the Standard solution and the Sample solution to separate 100-mL volumetric flasks, and transfer 2.0 mL of Solution A to a third volumetric flask to provide a blank. Dilute each flask with pH 9.6 Alkaline Borate Buffer (see Reagents, Indicators, and Solutions Solutions), and mix. Concomitantly determine the absorbances of the solutions in 1-cm cells at the wavelength of maximum absorbance at about 240 nm, with a suitable spectrophotometer, using the blank to set the instrument. Calculate the percentage of C10H15N2NaO3 in the Tablet: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 AU AS CS CU = absorbance of the solution from the Sample solution = absorbance of the solution from the Standard solution = concentration of USP Butabarbital RS in the Standard solution (mg/mL) = nominal concentration of butabarbital sodium in the Sample solution, based upon the labeled quantity/Tablet and the extent of dilution (mg/mL) rU rS CS
Mr2 = molecular weight of butabarbital, 212.25 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS Alcohol Determination, Method II 611: Between 95.0% and 115.0% of the labeled amount of C2H5OH ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Butabarbital RS
Butabarbital Sodium Tablets

(Comment on this Monograph)id=m10680=Butabarbital Sodium Tablets=B-Monos.pdf) DEFINITION Butabarbital Sodium Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of butabarbital sodium (C10H15N2NaO3). IDENTIFICATION INFRARED ABSORPTION 197K Sample: Equivalent to 150 mg of butabarbital sodium from powdered Tablets, in 1 mL of dimethyl sulfoxide and 1 mL of water. Add hydrochloric acid dropwise until the solution is just acid to litmus, and mix. Add 3 g of chromotographic siliceous earth, and mix. Proceed as directed for Chromatography 621, Column Partition Chromatography, packing the chromatographic tube as follows. The lower layer consists of 4 g of chromatographic siliceous earth mixed with 3 mL of sodium carbonate solution (1 in 10), and the upper layer is the Sample. Wash the column with 75 mL of a water-saturated mixture of isooctane and ether (4:1), and discard the washing. Elute the Butabarbital with 200 mL of water-saturated ether, collecting the eluate in a suitable vessel. Evaporate the eluate to dryness on a steam bath under a current of air, and dry the residue at 105 for 2 h. ASSAY PROCEDURE Internal standard solution: 1.2 mg/mL of secobarbital in chloroform Standard solution: 0.8 mg/mL of USP Butabarbital RS and 1 mg/mL of secobarbital in chloroform Sample solution: Transfer an equivalent to 50 mg of butabarbital sodium, from powdered Tablets (NLT 20), to a 50-mL volumetric flask. Add 35 mL of dilute ammonium hydroxide (1 in 25), dilute with water to volume, and mix. Filter, if necessary, discarding the first 15 mL of the filtrate, and transfer 25.0 mL of the clear solution to a separator. Add 2 mL of hydrochloric acid, and extract with three 25mL portions of chloroform. Filter the extracts through about 15 g of anhydrous sodium sulfate that is supported on a funnel by a small pledget of glass wool. Collect the combined filtrate in a 100-mL volumetric flask, wash the sodium sulfate with 15 mL of chloroform, collecting the washing with the filtrate, dilute with chloroform to volume, and mix. Combine 4.0 mL of this solution with 1.0 mL of Internal standard solution in a suitable container, and reduce the volume to about 1 mL by evaporation, with the aid of a stream of nitrogen, at room temperature. Chromatographic system and System suitability: Proceed as directed for Barbiturate Assay 361, Chromatographic System and System Suitability. Resolution: NLT 2.4 between butabarbital and secobarbital [NOTEThe relative retention times for butabarbital and secobarbital are 0.6 and 1.0, respectively.]
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Mr1 Mr2 = molecular weight of butabarbital sodium, 234.23 = molecular weight of butabarbital, 212.25
Official Monographs / Butalbital 153

Application volume: 10 L Developing solvent system: Acetone, dichloromethane, methanol, and ammonium hydroxide (5:3:1:1) Spray reagent: Dissolve 5 g of potassium hydroxide in a mixture of 25 mL of water and 75 mL of alcohol. Analysis Samples: Standard solutions A, B, C, and D and Sample solution In a suitable chromatographic chamber, lined with filter paper, place a volume of the Developing solvent system sufficient to develop the chromatogram. Cover the chamber, and allow it to equilibrate for 30 min. Apply 10 L each of the Sample solution and Standard solutions A, B, C, and D to a suitable thin-layer chromatographic plate. Develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and dry the plate in a current of air. Spray the plate with Spray reagent. Allow the plate to dry in warm air for 10 min, and examine the chromatograms under UV light. Acceptance criteria: The chromatograms show principal spots at the same RF value; and the sum of the intensities of any secondary spots, if present in the chromatogram from the Sample solution, are NMT 1% of that of the principal spot from Standard solution A. [NOTEThe relative intensities of the principal spots from Standard solution A, Standard solution B, Standard solution C, and Standard solution D are 1, 0.01, 0.005, and 0.0025, respectively.] SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 138141 LOSS ON DRYING 731: Dry a sample in a vacuum at room temperature to constant weight: it loses NMT 0.2% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Butalbital RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Butabarbital RS
Butalbital
(Comment on this Monograph)id=m10790=Butalbital=BMonos.pdf)
224.26 C11H16N2O3 2,4,6(1H,3H,5H)-Pyrimidinetrione, 5-(2-methylpropyl)-5-(2propenyl)-; 5-Allyl-5-isobutylbarbituric acid [77-26-9]. DEFINITION Butalbital contains NLT 98.0% and NMT 102.0% of C11H16N2O3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 246 nm Sample solution: 15 g/mL in 0.1 N sodium hydroxide Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 2.5% ASSAY PROCEDURE Sample: 180 mg of Butalbital Analysis: Dissolve in a mixture of 25 mL of alcohol and 25 mL of sodium carbonate solution (3 in 100). Titrate with 0.1 N silver nitrate VS, using a silver electrode, either with a suitable reference electrode containing a saturated aqueous solution of potassium nitrate, or a combination electrode in which the reference portion of the electrode contains a saturated aqueous solution of potassium nitrate. Each mL of 0.1 N silver nitrate is equivalent to 22.43 mg of C11H16N2O3. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Solution A: Chloroform and methanol (1:1) Sample solution: 40 mg/mL of Butalbital in Solution A Standard solution A: 40 mg/mL of USP Butalbital RS in Solution A Standard solution B: 0.4 mg/mL from Standard solution A in Solution A Standard solution C: Standard solution B and Solution A (1:1) Standard solution D: Standard solution C and Solution A (1:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture
Butalbital, Acetaminophen, and Caffeine Capsules

(Comment on this Monograph)id=m10798=Butalbital, Acetaminophen, and Caffeine Capsules=B-Monos.pdf) DEFINITION Butalbital, Acetaminophen, and Caffeine Capsules contain NLT 90.0% and NMT 110.0% of the labeled amounts of butalbital (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine (C8H10N4O2). IDENTIFICATION The retention times of the butalbital, acetaminophen, and caffeine peaks of the Sample solution correspond to those of the butalbital, acetaminophen, and caffeine peaks of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Transfer 800 mg of monobasic potassium phosphate to a 2000-mL volumetric flask. Dissolve in 1100 mL of water, and dilute with methanol to volume. Internal standard solution: 0.65 mg/mL of phenacetin in methanol Standard stock solution A: 0.01B mg/mL of USP Butalbital RS in Internal standard solution, B being the labeled amount
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the labeled amounts of acetaminophen, butalbital, and caffeine, respectively, in mg/Capsule Standard solution: Standard stock solution and water (1:19) Sample solution: Sample per Dissolution 711. Dilute with Medium to concentration that is similar to the Standard solution. Analysis Samples: Standard solution and Sample solution Pass a portion of the Sample solution through a filter of 10m or finer porosity. Separately inject equal volumes (20 L) of the filtrate and the Standard solution. Calculate the percentage of butalbital (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine (C8H10N4O2) dissolved by the same formula: Result = (rU/rS) (CS/CU) 100 = peak response of the corresponding analyte from the Sample solution = peak response of the corresponding USP rS Reference Standard from the Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of the corresponding CU analyte in the Sample solution (mg/mL) Tolerances: NLT 80% (Q) of the labeled amounts of C11H16N2O, C8H9NO2, and C8H10N4O2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Butalbital RS USP Caffeine RS
of butalbital in mg/Capsule, sonicating and shaking the solution, if necessary, to dissolve Standard stock solution B: 0.01C mg/mL of USP Caffeine RS in Internal standard solution, C being the labeled amount of caffeine in mg/Capsule, sonicating and shaking the solution, if necessary, to dissolve Standard solution: Transfer to a 50-mL volumetric flask 0.1A mg of USP Acetaminophen RS, A being the labeled amount of acetaminophen in mg/Capsule, 10.0 mL of Standard stock solution A, and 10.0 mL of Standard stock solution B. Sonicate for 5 min, and dilute with water to volume. This solution contains 0.002B mg/mL of butalbital, 0.002A mg/mL of acetaminophen, and 0.002C mg/mL of caffeine. Pass a portion of this solution through a suitable filter having a 0.5-m or finer porosity, and use the filtrate as the Standard solution. Sample stock solution: Equivalent to 1 average Capsule weight (powdered NLT 20 Capsules), in a 200-mL volumetric flask. Add Internal standard solution to volume. Sonicate for 15 min, mix, and allow to cool and settle. Sample solution: Transfer 20.0 mL of the clear supernatant of Sample stock solution to a 50-mL volumetric flask, and dilute with water to volume. Pass a portion of this solution through a filter of 0.5 m or finer porosity, discarding the first 5 mL of the filtrate. Use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 216 nm Column: 4-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen, caffeine, phenacetin, and butalbital are about 0.16, 0.33, 0.77, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.2 between any two peaks Column efficiency: NLT 1000 theoretical plates, calculated from the butalbital peak Relative standard deviation: NMT 2.0% of the acetaminophen, caffeine, and butalbital responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of butalbital (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine (C8H10N4O2) in the portion of Capsules taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the corresponding analyte to phenacetin from the Sample solution = peak response ratio of the corresponding analyte RS to phenacetin from the Standard solution CS = concentration of appropriate USP Reference Standard in the Standard solution (mg/mL) CU = nominal concentration of the corresponding analyte in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of C11H16N2O3, C8H9NO2, and C8H10N4O2 RU PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 60 min Mobile phase, Chromatographic system, and System suitability: Proceed as directed in the Assay. Standard stock solution: 0.02A mg/mL of USP Acetaminophen RS, 0.02B mg/mL of USP Butalbital RS, and 0.02C mg/mL of USP Caffeine RS, in which A, B, and C are
Butalbital, Acetaminophen, and Caffeine Tablets

(Comment on this Monograph)id=m10800=Butalbital, Acetaminophen, and Caffeine Tablets=B-Monos.pdf) DEFINITION Butalbital, Acetaminophen, and Caffeine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of butalbital (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine (C8H10N4O2). IDENTIFICATION The retention times of the butalbital, acetaminophen, and caffeine peaks of the Sample solution correspond to those of the butalbital, acetaminophen, and caffeine peaks of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Transfer 800 mg of monobasic potassium phosphate to a 2000-mL volumetric flask. Dissolve in 1100 mL of water, and dilute with methanol to volume. Internal standard solution: 0.65 mg/mL of phenacetin in methanol Standard stock solution A: 0.01B mg/mL of USP Butalbital RS in Internal standard solution, B being the labeled amount of butalbital, in mg/Tablet, sonicating and shaking the solution, if necessary, to dissolve Standard stock solution B: 0.01C mg/mL of USP Caffeine RS in Internal standard solution, C being the labeled amount of caffeine, in mg/Tablet, sonicating and shaking the solution, if necessary, to dissolve Standard solution: Transfer to a 50-mL volumetric flask 0.1A mg of USP Acetaminophen RS, A being the labeled
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amount of acetaminophen, in mg/Tablet, 10.0 mL of Standard stock solution A, and 10.0 mL of Standard stock solution B, sonicate for 5 min, and dilute with water to volume. This solution contains 0.002B mg/mL of butalbital, 0.002A mg/mL of acetaminophen, and 0.002C mg/mL of caffeine. Pass a portion of this solution through a suitable filter having a 0.5-m or finer porosity, and use the filtrate as the Standard solution. Sample stock solution: Equivalent to 1 average Tablet weight (powdered NLT 20 Tablets), into a 200-mL volumetric flask. Add Internal standard solution to volume. Sonicate for 15 min, mix, and allow to cool and settle. Sample solution: Transfer 20.0 mL of the clear supernatant of Sample stock solution to a 50-mL volumetric flask, and dilute with water to volume. Pass a portion of this solution through a filter of 0.5-m or finer porosity, discarding the first 5 mL of the filtrate. Use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 216 nm Column: 4-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetaminophen, caffeine, phenacetin, and butalbital are about 0.16, 0.33, 0.77, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.2 between any two peaks Column efficiency: NLT 1000 theoretical plates, calculated from the butalbital peak Relative standard deviation: NMT 2.0% of the acetaminophen, caffeine, and butalbital responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of butalbital (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine (C8H10N4O2) in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the corresponding analyte to phenacetin from the Sample solution RS = peak response ratio of the corresponding analyte to phenacetin from the Standard solution = concentration of appropriate USP Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of the corresponding CU analyte in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of C11H16N2O3, C8H9NO2, and C8H10N4O2 RU PERFORMANCE TESTS DISSOLUTION, ANALYSIS FOR A POOLED SAMPLE 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Mobile phase, Chromatographic system, and System suitability: Proceed as directed in the Assay, except use an injection size of 20 L for the Analysis. Standard stock solution: 0.02A mg/mL of USP Acetaminophen RS, 0.02B mg/mL of USP Butalbital RS, and 0.02C mg/mL of USP Caffeine RS, in which A, B, and C are the labeled amounts of acetaminophen, butalbital, and caffeine, respectively, in mg/Tablet Standard solution: Standard stock solution and water (1:19) Sample solution: Sample per Dissolution 711. Dilute with Medium as needed and pass through a filter of 10 m or finer porosity.

Analysis Samples: Standard solution and Sample solution Calculate the percentage of butalbital (C11H16N2O3), acetaminophen (C8H9NO2), and caffeine (C8H10N4O2) dissolved by the same formula: Result = (rU/rS) CS V 100/L = peak response of the corresponding analyte from the Sample solution = peak response of the corresponding USP rS Reference Standard from the Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) V = volume of Medium, 900 mL L = Tablet label claim of corresponding analyte (mg) Tolerances: NLT 80% (Q) of the labeled amounts of C11H16N2O, C8H9NO2, and C8H10N4O2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Acetaminophen RS USP Butalbital RS USP Caffeine RS
Butalbital and Aspirin Tablets

(Comment on this Monograph)id=m10805=Butalbital and Aspirin Tablets=B-Monos.pdf) DEFINITION Butalbital and Aspirin Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of butalbital (C11H16N2O3) and aspirin (C9H8O4). IDENTIFICATION The retention times of the butalbital and aspirin peaks of the Sample solution correspond to those of the butalbital and aspirin peaks of Standard solution A and Standard solution B, as obtained in the Assay. ASSAY BUTALBITAL AND ASPIRIN Mobile phase: Acetonitrile, phosphoric acid, and water (725:4:3100). Adjust the ratio as necessary. Solvent mixture: Formic acid and acetonitrile (1:100) Standard stock solution A: 3250J g/mL of USP Butalbital RS in Solvent mixture, J being the ratio of the labeled amount, in mg, of butalbital to the labeled amount of aspirin, in mg/Tablet Standard stock solution B: 200 g/mL of USP Salicylic Acid RS in Solvent mixture Standard solution A: 25.0 mL of Standard stock solution A and 3.0 mL of Standard stock solution B to a 250-mL volumetric flask. Dilute with Solvent mixture to volume. This solution contains 325J g of butalbital and 2.4 g of salicylic acid/mL. Standard solution B: 325 g/mL of USP Aspirin RS in Solvent mixture System suitability solution: 4.0 mL of Standard stock solution A and 3.0 mL of Standard stock solution B to a 50mL volumetric flask. Dilute with Solvent mixture to volume. Sample solution: Equivalent to 320 g/mL of aspirin from powdered Tablets (NLT 20 Tablets), in Solvent mixture. Sonicate for 15 min, and mix. Pass a portion of this solution through a fliter of 0.5-m porosity before use. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 214 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 1.5 mL/min Injection size: 1025 L System suitability Sample: Standard solution [NOTEThe relative retention times for aspirin, salicylic acid, and butalbital are about 0.6, 0.85, and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the butalbital and salicylic acid peaks Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution [NOTEFilter a portion of the Sample solution through a 0.5-m filter. After use, the column may be regenerated by passing through it at least 50 mL of a mixture of acetonitrile, methanol, and water (1:1:1), followed by 50 mL of a mixture of acetonitrile and water (1:1).] Calculate the percentage of C11H16N2O3 dissolved: Result = (rU/rS) (CS/CU) 100 = peak response of butalbital from the Sample solution = peak response butalbital from the Standard rS solution CS = concentration of USP Butalbital RS in the Standard solution (g/mL) CU = nominal concentration of butalbital in the Sample solution (g/mL) Determination of dissolved aspirin Solution A: 5.98 g of sodium acetate trihydrate in 500 mL of water. Add 2.5 mL of glacial acetic acid, dilute with water to 1000 mL, and mix. Adjust this solution with glacial acetic acid to a pH of 4.5 0.05. Sample solution: Solution under test diluted with 4 volumes of Solution A Standard solution: A known concentration of USP Aspirin RS diluted with 4 volumes of Solution A [NOTEPrepare the Standard solution at the time of use. An amount of alcohol not to exceed 1% of the total volume of the Standard solution may be used to bring the Reference Standard into solution before dilution first with water and then with 4 volumes of Solution A.] Analysis: Determine the amount of aspirin (C9H8O4) dissolved from UV absorbances at the wavelength of the isosbestic point of aspirin and salicylic acid at 265 2 nm of filtered portions of the Sample solution in comparison with the Standard solution. Tolerances: NLT 75% (Q) of the labeled amounts of butalbital (C11H16N2O3) and aspirin (C9H8O4) is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to butalbital and for Weight Variation with respect to aspirin. rU IMPURITIES Organic Impurities PROCEDURE: Limit of Free Salicylic Acid Mobile Phase, Solvent mixture, Standard stock solution A, Standard stock solution B, Standard solution A, System suitability solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed under Assay
Mode: LC Detector: UV 214 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: System suitability solution, Standard solution A, and Standard solution B [NOTEThe relative retention times for aspirin, salicylic acid, and butalbital are about 0.6, 0.85, and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the butalbital and salicylic acid peaks, System suitability solution Relative standard deviation: NMT 3.0% for butalbital and aspirin, and NMT 6.0% for salicylic acid, Standard solutions A, and B Analysis Samples: Standard solution A, Standard solution B, and Sample solution [NOTEAfter use, the column may be regenerated by passing through it at least 50 mL of a mixture of acetonitrile, methanol, and water (1:1:1), followed by a mixture of acetonitrile and water (1:1).] Calculate the percentage of C11H16N2O3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of butalbital from the Sample solution = peak response of butalbital from Standard rS solution A = concentration of USP Butalbital RS in Standard CS solution A (g/mL) = nominal concentration of butalbital in the CU Sample solution (g/mL) Calculate the percentage of C9H8O4 in the portion of Tablets taken: Result = (rU /rS) (CS/CU) 100 = peak response of aspirin from the Sample solution = peak response of aspirin from Standard solution B = concentration of USP Aspirin RS in Standard solution B (g/mL) = nominal concentration of aspirin in the Sample CU solution (g/mL) Acceptance criteria: 90.0%110.0% of the labeled amounts of C11H16N2O3 and C9H8O4 rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 60 min Determination of dissolved butalbital Mobile phase: Acetonitrile, phosphoric acid, and water (725:4:3100). Adjust the ratio as necessary. Sample solution: Sample per Dissolution 711. Dilute with Medium to concentration that is similar to the Standard solution. Standard solution: 1 g/mL of USP Butalbital RS for each mg of the labeled amount/Tablet and 30 g/mL of salicylic acid in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
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Analysis Samples: Standard solution and Sample solution Calculate the percentage of free salicylic acid in the Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of salicylic acid from the Sample solution = peak response of salicylic acid from Standard rS solution A = concentration of USP Salicylic Acid RS in CS Standard solution A = nominal concentration of aspirin in the Sample CU solution Acceptance criteria: NMT 3.0% of free salicylic acid rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Butalbital RS USP Salicylic Acid RS

Mode: LC Detector: Set at the wavelength of the isosbestic point of aspirin and salicylic acid at 277 and 210 nm. Column: 3.9-mm 30-cm; packing L1 Temperature: 35 1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard solution A and Standard solution B [NOTEThe relative retention times for caffeine, aspirin, and butalbital are about 0.45, 0.6, and 1.0, respectively, in Standard solution A.] [NOTEThe salicylic acid peak has the same retention time as that of the aspirin peak obtained in the chromatogram of Standard solution B. If the retention time of the salicylic acid peak differs from that of the aspirin peak, adjust the pH of the Mobile phase with 0.2 N potassium hydroxide or 1 M phosphoric acid so that the salicylic acid peak has the same retention time as that of the aspirin peak. The retention time of the salicylic acid peak decreases 0.3 min for each 0.1 pH increase. The retention time of the aspirin peak is essentially unaffected by such pH adjustments.] Suitability requirements Resolution: NLT 2.0 between the caffeine and aspirin peaks, Standard solution A Column efficiency: NLT 2000 theoretical plates from the butalbital peak, Standard solution A Relative standard deviation: NMT 2.0% of the caffeine, aspirin, and butalbital responses, Standard solution A Analysis Samples: Standard solution A and Sample solution Record the chromatograms, using the 277-nm detector to record the caffeine and aspirin peak responses and the 210-nm detector to record the butalbital peak responses Calculate the percentage of caffeine (C8H10N4O2), aspirin (C9H8O4), and butalbital (C11H16 N2O3) in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response of the corresponding analyte from the Sample solution rS = peak response of the corresponding USP Reference Standard from the Standard solution A CS = concentration of appropriate USP Reference Standard in Standard solution A (mg/mL) CU = nominal concentration of the corresponding analyte in the Sample solution (mg/mL) Correct the amount of aspirin obtained for the amount of salicylic acid present by the formula: rU Result = A (0.01 F A) = quantity of aspirin in the portion of Capsules taken to prepare the Sample solution (mg) F = percentage of salicylic acid obtained in the Procedure for Limit of free salicylic acid Acceptance criteria: 90.0%110.0% of C11H16 N2O3, C9H8O4, and C8H10N4O2 PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 1000 mL Apparatus 2: 50 rpm Time: 60 min Sample solution: Samples per Dissolution 711. Solution A, Solution B, Mobile phase, Standard stock solution, Standard solution A, Standard solution B, A
Butalbital, Aspirin, and Caffeine Capsules

(Comment on this Monograph)id=m10810=Butalbital, Aspirin, and Caffeine Capsules=B-Monos.pdf) DEFINITION Butalbital, Aspirin, and Caffeine Capsules contain NLT 90.0% and NMT 110.0% of the labeled amounts of butalbital (C11H16 N2O3), aspirin (C9H8O4), and caffeine (C8H10N4O2). IDENTIFICATION The retention times of the butalbital, aspirin, and caffeine peaks of the Sample solution correspond to those of the butalbital, aspirin, and caffeine peaks of the Standard solution A, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.361 mg/mL of monobasic potassium phosphate Solution B: Methanol and Solution A (9:11). Adjust with phosphoric acid to a pH of 2.5 0.05 Mobile phase: Methanol and Solution A (9:11). Adjust with phosphoric acid to a pH of 3.9. Standard stock solution: 1.6 mg/mL of USP Aspirin RS in Solution B, sonicating and shaking the solution, if necessary, to dissolve. Use this solution within 24 h. Standard solution A: 1.6J mg/mL of USP Butalbital RS and 1.6J mg/mL of USP Caffeine RS in Standard stock solution, J and J being the ratios of the respective labeled amounts, in mg, of butalbital and caffeine to the labeled amount, in mg/Capsule of aspirin, sonicating and shaking the solution, if necessary, to dissolve. Use this solution within 24 h. Standard solution B: 0.1 mg/mL of salicylic acid in Solution B. Pass this solution through a suitable filter of 0.5-m or finer porosity. Sample solution: Equivalent to 325 mg of aspirin (remove, as completely as possible, the contents of NLT 20 Capsules), into a 200-mL volumetric flask. Dilute with Solution B to volume, sonicate for 30 min, and mix. Pass a portion of this solution through a suitable filter of 0.5-m or finer porosity, and use the filtrate. Use this solution within 24 h. [NOTE Reserve the remaining portion of the powder for the Procedure for Limit of Free Salicylic Acid] Chromatographic system (See Chromatography 621, System Suitability.)
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IDENTIFICATION The retention times of the butalbital, aspirin, and caffeine peaks of the Sample solution correspond to those of the butalbital, aspirin, and caffeine peaks of the Standard solution A, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.361 mg/mL of monobasic potassium phosphate Solution B: Methanol and Solution A (9:11). Adjust with phosphoric acid to a pH of 2.5 0.05 Mobile phase: Methanol and Solution A (9:11). Adjust with phosphoric acid to a pH of 3.9. Standard stock solution: 1.6 mg/mL of USP Aspirin RS in Solution B, sonicating and shaking the solution, if necessary, to dissolve. Use this solution within 24 h. Standard solution A: 1.6J mg/mL of USP Butalbital RS and 1.6J mg/mL of USP Caffeine RS in Standard stock solution, J and J being the ratios of the respective labeled amounts, in mg, of butalbital and caffeine to the labeled amount, in mg/Capsule of aspirin, sonicating and shaking the solution, if necessary, to dissolve. Use this solution within 24 h. Standard solution B: 0.1 mg/mL of salicylic acid in Solution B. Pass this solution through a suitable filter of 0.5-m or finer porosity. Sample solution: Equivalent to 325 mg of aspirin from powdered Tablets (NLT 20 Tablets), into a 200-mL volumetric flask. Dilute with Solution B to volume, and sonicate for 30 min. Pass a portion of this solution through a suitable filter of 0.5-m or finer porosity, and use the filtrate. Use this solution within 24 h. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Set at the wavelength of the isosbestic point of aspirin and salicylic acid at 277 and 210 nm. Column: 3.9-mm 30-cm; packing L1 Temperature: 35 1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution A and Standard solution B [NOTEThe relative retention times for caffeine, aspirin, and butalbital are about 0.45, 0.6, and 1.0, respectively, in Standard solution A.] [NOTEThe salicylic acid peak has the same retention time as that of the aspirin peak obtained in the chromatogram of the Standard solution B. If the retention time of the salicylic acid peak differs from that of the aspirin peak, adjust the pH of the Mobile phase with 0.2 N potassium hydroxide or 1 M phosphoric acid so that the salicylic acid peak has the same retention time as that of the aspirin peak. The retention time of the salicylic acid peak decreases 0.3 min for each 0.1 pH increase. The retention time of the aspirin peak is essentially unaffected by such pH adjustments.] Suitability requirements Resolution: NLT 2.0 between the caffeine and aspirin peaks, Standard solution A Column efficiency: NLT 2000 theoretical plates from the butalbital peak, Standard solution A Relative standard deviation: NMT 2.0% of the caffeine, aspirin, and butalbital responses, Standard solution A Analysis Samples: Standard solution A and Sample solution Record the chromatograms, using the 277-nm detector to record the caffeine and aspirin peak areas and the 210-nm detector to record the butalbital peak areas.
Chromatographic system, System suitability, and Analysis: Proceed as directed in the Assay. Tolerances: NLT 75% (Q) of the labeled amounts of butalbital (C11H16N2O3), caffeine (C8H10N4O2), and aspirin (C9H8O4) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID [NOTEUse glassware in this test.] Solution A: Phosphoric acid and methanol (1:999) Standard solution: 0.0012 mg/mL of USP Salicylic Acid RS in Solution A. [NOTEUse this solution promptly.] Sample solution: Equivalent to 65 mg of aspirin from a portion of the powder remaining from the solution of the Sample solution in the Assay, to a 200-mL flask, add 100.0 mL of Solution A, and shake for 1 min. Promptly filter a portion of this solution, discarding the first 15 mL of the filtrate, and use the clear filtrate. Use this solution within 20 min after the addition of the Solution A. [NOTEPerform this test on the same day the powder is removed from the Capsules] Fluorimetric conditions Excitation wavelength: 305 nm Emmission wavelength: 444 nm Analysis Samples: Standard solution and Sample solution Allow the Samples to equilibrate for 2 min in the fluorimeter. Calculate the percentage of salicylic acid in the portion of Capsules taken: Result = (IU/IS) (CS/CU) 100 = fluorescence intensity readings from the Sample solution = fluorescence intensity readings from the IS Standard solution = concentration of USP Salicylic Acid RS in the CS Standard solution (mg/mL) = nominal concentration of aspirin in the Sample CU solution (mg/mL) [NOTEIf the intensity of the Sample solution greatly exceeds that of the Standard solution, immediately transfer 5.0 mL of the Sample solution to a 50-mL volumetric flask, dilute with Solution A to volume, and mix. Immediately determine the intensity of this solution. Calculate the percentage of salicylic acid in the portion of Capsules taken, by using the same formula.] Acceptance criteria: NMT 2.5% IU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Butalbital RS USP Caffeine RS USP Salicylic Acid RS
Butalbital, Aspirin, and Caffeine Tablets

(Comment on this Monograph)id=m10820=Butalbital, Aspirin, and Caffeine Tablets=B-Monos.pdf) DEFINITION Butalbital, Aspirin, and Caffeine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of butalbital (C11H16N2O3), aspirin (C9H8O4), and caffeine (C8H10N4O2).
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Calculate the percentage of butalbital (C11H16N2O3), aspirin (C9H8O4), and caffeine (C8H10N4O2) in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of the corresponding analyte from the Sample solution = peak response of the corresponding USP rS Reference Standard from the Standard solution A = concentration of appropriate USP Reference CS Standard in Standard solution A (mg/mL) = nominal concentration of the corresponding CU analyte in the Sample solution (mg/mL) Correct the amount of aspirin obtained for the amount of salicylic acid present by the formula: Result = A (0.01 F A) = quantity of aspirin in the portion of Tablets taken to prepare the Sample solution (mg) F = percentage of salicylic acid obtained in the Procedure for Limit of Free Salicylic Acid Acceptance criteria: 90.0%110.0% of C11H16 N2O3, C9H8O4, and C8H10N4O2 PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 60 min Sample solution Sample per Dissolution 711 Solution A, Solution B, Mobile phase, Standard stock solution, Standard solution A, Standard solution B, Chromatographic system, System suitability, and Analysis: Proceed as directed under Assay. Tolerances: NLT 80% (Q) of the labeled amounts of butalbital (C11H16N2O3), caffeine (C8H10N4O2), and aspirin (C9H8O4) is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID [NOTEUse glassware in this test.] Solution A: Phosphoric acid and methanol (1:999) Standard solution: 0.0012 mg/mL of USP Salicylic Acid RS in Solution A [NOTEUse this solution promptly.] Sample solution: Equivalent to 65 mg of aspirin from powdered Tablets (NLT 20 Tablets), to a 200-mL flask. Add 100.0 mL of Solution A, and shake by mechanical means for 15 min. Filter a portion of this solution, discarding the first 15 mL of the filtrate, and use the clear filtrate. Use this solution within 20 min after the addition of the Solution A. [NOTEPerform this test on the same day the Tablets are powdered.] Fluorimetric conditions Excitation wavelength: 305 nm Emmission wavelength: 444 nm Analysis Samples: Standard solution and Sample solution Allow the Samples to equilibrate for 2 min in the fluorimeter. Calculate the percentage of salicylic acid in the portion of Tablets taken: Result = (IU/IS) (CS/IU) 100 IU = fluorescence intensity readings from the Sample solution A IS

= fluorescence intensity readings from the Standard solution = concentration of USP Salicylic Acid RS in the CS Standard solution (mg/mL) = nominal concentration of aspirin in the Sample CU solution (mg/mL) [NOTEIf the intensity of the Sample solution greatly exceeds that of the Standard solution, immediately transfer 5.0 mL of the Sample solution to a 50-mL volumetric flask, dilute with Solution A to volume, and mix. Immediately determine the intensity of this solution, and calculate the percentage of salicylic acid in the portion of Tablets taken, by using the same formula.] Acceptance criteria: NMT 3.0% is found ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Butalbital RS USP Caffeine RS USP Salicylic Acid RS
Butalbital, Aspirin, Caffeine, and Codeine Phosphate Capsules

(Comment on this Monograph)id=m10825=Butalbital, Aspirin, Caffeine, and Codeine Phosphate Capsules=B-Monos.pdf) DEFINITION Butalbital, Aspirin, Caffeine, and Codeine Phosphate Capsules contain NLT 90.0% and NMT 110.0% of the labeled amounts of butalbital (C11H16N2O3), aspirin (C9H8O4), caffeine (C8H10N4O2), and codeine phosphate (C18H21 NO3 H3PO4 1 /2H2O). IDENTIFICATION The retention times of the butalbital, aspirin, caffeine, and codeine peaks of the Sample solution correspond to those of the butalbital, aspirin, caffeine, and codeine peaks of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.361 mg/mL of monobasic potassium phosphate Solution B: Methanol and Solution A (9:11). Adjust with phosphoric acid to a pH of 2.5 0.05. Mobile phase: Methanol and Solution A (9:11). Adjust with phosphoric acid to a pH of 3.9. Standard stock solution A: 1.6 mg/mL of USP Aspirin RS in Solution B, sonicating and shaking the solution, if necessary, to dissolve. Use this solution within 24 h. Standard solution A: 1.6J mg/mL of USP Butalbital RS, 1.6J mg/mL of USP Caffeine RS, and 1.6J mg/mL of USP Codeine Phosphate RS in Standard stock solution A, J, J, and J being the ratios of the respective labeled amounts, in mg, of butalbital, caffeine, and codeine phosphate to the labeled amount, in mg/Capsule of aspirin, sonicating and shaking the solution, if necessary, to dissolve. Standard solution B: 0.1 mg/mL of salicylic acid in Solution B. Pass this solution through a suitable filter of 0.5-m or finer porosity. Sample solution: Equivalent to 325 mg of aspirin (remove, as completely as possible, the contents of NLT 20 Capsules), into a 200-mL volumetric flask. Dilute with Solution B to volume, sonicate for 30 min, and mix. Pass a portion of this solution through a suitable filter of 0.5-m or finer porosity, and use the filtrate. Use this solution within 24 h. [NOTE
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Correct the amount of aspirin obtained for the amount of salicylic acid present taken: Result = A (0.01 F A) = quantity of aspirin in the portion of Capsules taken to prepare the Sample solution (mg) F = percentage of salicylic acid obtained in the Procedure for Limit of Free Salicylic Acid Acceptance criteria: 90.0%110.0% of C11H16N2O3, C9H8O4, C8H10N4O2, and C18H21 NO3 H3PO4 1/2H2O PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 1000 mL Apparatus 2: 50 rpm Time: 60 min Solution A, Solution B, Mobile phase, and Standard solution B: Prepare as directed in the Assay. Standard solution C: Transfer 5.0 mL of Standard solution B, prepared as directed in the Assay, to a 50-mL volumetric flask, and dilute with Solution B to volume. Standard stock solution A: 0.16 mg/mL of USP Aspirin RS in a mixture of Solution B and Medium (1:1). Use this solution within 24 h. Standard solution A: 0.16J mg/mL of USP Butalbital RS, 0.16J mg/mL of USP Caffeine RS, and 0.16J mg/mL of USP Codeine Phosphate RS in Standard stock solution A, J, J, and J being the ratios of the respective labeled amounts, in mg, of butalbital, caffeine, and codeine phosphate to the labeled amount of aspirin, in mg/Capsule, sonicating and shaking the solution, if necessary to dissolve. Pass a portion of this solution through a suitable filter of 0.5-m or finer porosity. Use this solution within 24 h. Sample solution: Pass 20 mL of the solution under test through a suitable filter of 0.5-m or finer porosity, discarding the first 2 mL of the filtrate. Mix 5.0 mL of the filtrate and 5.0 mL of Solution B. Chromatographic system: Proceed as directed in the Assay, except to inject 100 L, instead of 10 L, into the chromatograph, and use Standard solution C instead of Standard solution B. System suitability: Proceed as directed under Assay. Analysis Samples: Standard solution A and Sample solution Use the 277-nm detector to record the caffeine and aspirin peaks and the 210-nm detector to record the butalbital and codeine responses. Calculate the percentage of caffeine (C8H10N4O2), aspirin (C9H8O4), and butalbital (C11H16N2O3) dissolved: Result = (rU/rS) (CS/CU) 100 rU = peak response of the corresponding analyte from the Sample solution rS = peak response of the corresponding analyte from Standard solution A CS = concentration of the appropriate USP Reference Standard in Standard solution A (mg/mL) = nominal concentration of the corresponding CU analyte in the Sample solution (mg/mL) Calculate the percentage of codeine phosphate (C18H21NO3 H3PO4 1/2H2O) dissolved: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS = peak response of codeine from the Sample solution = peak response of codeine from Standard solution A = concentration of USP Codeine Phosphate RS in Standard solution A (mg/mL) A
Reserve the remaining portion of the powder for the Procedure for Limit of Free Salicylic Acid] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Set at the wavelength of the isosbestic point of aspirin and salicylic acid at 277 and 210 nm. Column: 3.9-mm 30-cm; packing L1 Temperature: 35 1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution A and Standard solution B [NOTEThe relative retention times for codeine, caffeine, aspirin, and butalbital are about 0.3, 0.45, 0.6, and 1.0, respectively.] [NOTEThe salicylic acid peak has the same retention time as that of the aspirin peak obtained in the chromatogram of the Standard solution B. If the retention time of the salicylic acid peak differs from that of the aspirin peak, adjust the pH of the Mobile phase with 0.2 N potassium hydroxide or 1 M phosphoric acid so that the salicylic acid peak has the same retention time as that of the aspirin peak. The retention time of the salicylic acid peak decreases 0.3 min for each 0.1 pH increase. The retention time of the aspirin peak is essentially unaffected by such pH adjustments.] Suitability requirements Resolution: NLT 2.0 between the caffeine and aspirin peaks, Standard solution A Column efficiency: NLT 2000 theoretical plates from the butalbital peak, Standard solution A Relative standard deviation: NMT 2.0% of the codeine, caffeine, aspirin, and butalbital responses, Standard solution A Analysis Samples: Standard solution A and Sample solution Record the chromatograms, using the 277-nm detector to record the caffeine and aspirin peak responses and the 210-nm detector to measure the codeine and butalbital responses. Calculate the percentage of caffeine (C8H10N4O2), aspirin (C9H8O4), and butalbital (C11H16N2O3) in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of the corresponding analyte from the Sample solution rS = peak response of the corresponding USP Reference Standard from Standard solution A = concentration of appropriate USP Reference CS Standard in Standard solution A (mg/mL) = nominal concentration of the corresponding CU analyte in the Sample solution (mg/mL) Calculate the percentage of codeine phosphate (C18H21NO3 H3PO4 1/2H2O) in the portion of Capsules taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 Mr2 = peak response of codeine from the Sample solution = peak response of codeine from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of codeine phosphate in the Sample solution (mg/mL) = molecular weight of codeine phosphate hemihydrate, 406.37 = molecular weight of anhydrous codeine phosphate, 397.37
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= nominal concentration of codeine in the Sample solution (mg/mL) Mr1 = molecular weight of codeine phosphate hemihydrate, 406.37 Mr2 = molecular weight of anhydrous codeine phosphate, 397.37 Tolerances: NLT 75% (Q) of the labeled amounts of butalbital (C11H16N2O3), aspirin (C9H8O4), caffeine (C8H10N4O2), and codeine phosphate (C18H21NO3 H3PO4 1 /2H2O) is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID [NOTEUse glassware in this test.] Solution A: Phosphoric acid and methanol (1:999) Standard solution: 0.0012 mg/mL of USP Salicylic Acid RS in Solution A [NOTEUse this solution promptly.] Sample solution: Equivalent to 65 mg of aspirin from a portion of the powder remaining from the solution of the Sample solution in the Assay, to a 200-mL flask. Add 100.0 mL of Solution A, and shake for 1 min. Promptly filter a portion of this solution, discarding the first 15 mL of the filtrate, and use the clear filtrate. Use this solution within 20 min after the addition of the Solution A. [NOTEPerform this test on the same day the powder is removed from the Capsules.] Fluorimetric conditions Excitation wavelength: 305 nm Emmission wavelength: 444 nm Analysis Samples: Standard solution and Sample solution Allow to equilibrate for 2 min in the fluorimeter. Calculate the percentage of salicylic acid in the portion of Capsules taken: Result = (IU/IS) (CS/CU) 100 = fluorescence intensity readings from the Sample solution IS = fluorescence intensity readings from the Standard solution CS = concentration of USP Salicylic Acid RS in the Standard solution (mg/mL) CU = nominal concentration of aspirin in the Sample solution (mg/mL) [NOTEIf the intensity of the Sample solution greatly exceeds that of the Standard solution, immediately transfer 5.0 mL of the Sample solution to a 50-mL volumetric flask, dilute with Solution A to volume, and mix. Immediately determine the intensity of this solution. Calculate the percentage of salicylic acid in the portion of Capsules taken.] Acceptance criteria: NMT 3.0% IU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Butalbital RS USP Caffeine RS USP Codeine Phosphate RS USP Salicylic Acid RS CU
Official Monographs / Butamben 161
Butamben
(Comment on this Monograph)id=m10840=Butamben=BMonos.pdf)
C11H15NO2 Benzoic acid, 4-amino-, butyl ester; Butyl p-aminobenzoate [94-25-7].
193.24
DEFINITION Butamben, dried over phosphorus pentoxide for 3 h, contains NLT 98.0% and NMT 101.0% of C11H15NO2. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Solution A: Dissolve, without warming, 0.5 g of ferrocyphen in 50 mL of sulfuric acid. Sample solution: Dissolve 400 mg of Butamben, previously dried, in a mixture of 100 mL of water and 20 mL of hydrochloric acid. Add 1 mL of Solution A, and cool the solution in an ice bath to 10. Analysis: Titrate the Sample solution with 0.1 M sodium nitrite VS to a violet endpoint that is stable for NLT 3 min. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 M sodium nitrite is equivalent to 19.32 mg of C11H15NO2. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% CHLORIDE AND SULFATE, Chloride 221: To a solution of 200 mg in 10 mL of alcohol, add 1 mL of 2 N nitric acid and a few drops of silver nitrate TS: no opalescence is produced. HEAVY METALS, Method I 231: Dissolve 2 g in 2 mL of 1 N acetic acid and sufficient alcohol to make 25 mL: NMT 10 ppm SPECIFIC TESTS COMPLETENESS AND COLOR OF SOLUTION: One g dissolves completely in 30 mL of alcohol and in 30 mL of ether, and the solutions are colorless. MELTING RANGE OR TEMPERATURE, Class I 741: 5759 REACTION: Dissolve 1 g in 10 mL of neutralized alcohol: a clear solution results. Dilute with 10 mL of water this solution, and add 2 drops of phenolphthalein TS and 1 drop of 0.1 N sodium hydroxide: a red color is produced. LOSS ON DRYING 731: Dry it over phosphorus pentoxide for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Butamben RS
162
Butoconazole / Official Monographs
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Developing solvent system: Chloroform, tetrahydrofuran, cyclohexane, and ammonium hydroxide (18:18:13:1) Visualization: 22 Acceptance criteria: Any spot of the Sample solution, except for the prinicpal spot, is not more intense than the principal spot of the Standard solution, 0.01 mg/mL. NMT 2.0% of total impurities is found. SPECIFIC TESTS LOSS ON DRYING 731: Dry a sample in a vacuum at 60 for 3 h: it loses NMT 1.0% of its weight.
Butoconazole Nitrate
(Comment on this Monograph)id=m10900=Butoconazole Nitrate=B-Monos.pdf)
C19H17Cl3N2S HNO3 1H-Imidazole, 1-[4-(4-chlorophenyl)-2-[(2,6dichlorophenyl)thio]butyl-, mononitrate, ()-; ()-1-[4-(p-Chlorophenyl)-2-[(2,6-dichlorophenyl)thio]butyl]imidazole mononitrate [64872-77-1].
474.79
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Butoconazole Nitrate RS
DEFINITION Butoconazole Nitrate contains NLT 98.0% and NMT 102.0% of C19H17Cl3N2S HNO3, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Buffer: Monobasic potassium phosphate 2.18 g/L and dibasic potassium phosphate 4.18 g/L in water Mobile phase: Methanol and Buffer (3:1) Standard solution: USP Butoconazole Nitrate RS 0.2 mg/mL in Mobile phase Sample solution: Butoconazole Nitrate 0.2 mg/mL in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 229 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 40 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2800 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H17Cl3N2S HNO3 in the portion of Butoconazole Nitrate taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Butoconazole Nitrate RS in the Standard solution (mg/mL) = concentration of Butoconazole Nitrate in the CU Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Standard solution: 0.01, 0.05, 0.1, and 0.2 mg/mL in methylene chloride and methanol (2:1) Sample solution: 10 mg/mL in methylene chloride and methanol (2:1) rU rS CS
Butoconazole Nitrate Vaginal Cream

(Comment on this Monograph)id=m10904=Butoconazole Nitrate Vaginal Cream=B-Monos.pdf) DEFINITION Butoconazole Nitrate Vaginal Cream contains Butoconazole Nitrate in a suitable cream base. It contains NLT 90.0% and NMT 110.0% of the labeled amount of butoconazole nitrate (C19H17Cl3N2S HNO3). IDENTIFICATION PROCEDURE Analysis: Prepare a mixture of the Standard solution and the Sample solution (1:1), prepared as directed in the Assay, and chromatograph as directed in the Assay. Acceptance criteria: The chromatogram so obtained exhibits two main peaks, corresponding to butoconazole nitrate and the internal standard. ASSAY PROCEDURE Buffer: Potassium acetate 1.4 g in 980 mL of water. Adjust with about 2 mL of glacial acetic acid to a pH of 4.3 0.1, dilute with water to 1000 mL, and mix. Adjust the buffer molarity (0.0180.072 M) as necessary to obtain suitable chromatographic performance. Increased retention time may be achieved by a decrease in the buffer molarity. Diluent: Methanol and Buffer (3:2) Mobile phase: Methanol and Buffer (13:7) Internal standard solution: 1-Benzylimidazole 1.6 mg/mL in methanol Standard stock solution: USP Butoconazole Nitrate RS 0.4 mg/mL in methanol Standard solution: Transfer 2.0 mL of Standard stock solution and 3.0 mL of Internal standard solution to a 50-mL flask, and add 35.0 mL of Diluent. Sample stock solution: Sonicate a quantity of Butoconazole. Sample solution: Transfer 2.0 mL of Sample stock solution and 3.0 mL of Internal standard solution to a 50-mL flask, and add 35.0 mL of Diluent. Allow the precipitated excipients that form to rise to the top of the solution, remove them by aspiration, and discard. Centrifuge or filter the remaining solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 225 nm Column: 4.6-mm 25-cm; packing L9 that has been converted to the potassium form by the use of 0.555 M potassium acetate solution
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Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for butoconazole nitrate and 1-benzylimidazole are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between the analyte and internal standard peaks Column efficiency: NLT 1100 theoretical plates Tailing factor: NMT 2.1 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H17Cl3N2S HNO3 in the portion of Vaginal Cream taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of butoconazole nitrate to the internal standard from the Sample solution = peak response ratio of butoconazole nitrate to RS the internal standard from the Standard solution = concentration of USP Butoconazole Nitrate RS in CS the Standard solution (g/mL) = nominal concentration of butoconazole nitrate in CU the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers. Avoid excessive heat and avoid freezing. USP REFERENCE STANDARDS 11 USP Butoconazole Nitrate RS RU
Official Monographs / Butorphanol 163

Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 47.76 mg of C21H29NO2 C4H6O6. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method III 231: NMT 30 ppm Organic Impurities PROCEDURE 1 Standard solution: 1 mg/mL of USP Butorphanol Tartrate RS in methanol Sample solution: 10 mg/mL of Butorphanol Tartrate in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 50 L of Sample solution, 5 L and 10 L of Standard solution Developing solvent system: Chloroform, methanol, benzene, and ammonium hydroxide (17:5:4:1) Spray reagent: Prepare a 1in10 solution of chloroplatinic acid in water. To 0.5 mL of this solution, add 33 mL of water and 1 g of potassium iodide. Prepare fresh daily. Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Place the plate in a developing chamber containing, and equilibrated with the Developing solvent system. Develop the chromatogram until the solvent front has moved 10 cm above the line of application. Remove the plate, mark the solvent front, and allow the solvent to evaporate. Spray the plate with Spray reagent. Estimate the percentage of the impurities present in the Sample solution by comparing the intensities of secondary spots, if present, with the intensities of the principal spots of the Standard solutions. Acceptance criteria: The sum of the impurities observed is NMT 2.0%. PROCEDURE 2 Sample solution: 10 mg/mL of Butorphanol Tartrate in methanol Chromatographic system Mode: GC Detector: Flame ionization Column: 1.8-m 4-mm; glass column containing 3% liquid phase G3 on support S1AB Temperature Injection port: 280 Column: 250 Detector: 290 Carrier gas: Nitrogen Injection size: 1 L Analysis Sample: Sample solution [NOTEThe retention time for the alpha isomer of butorphanol tartrate, for butorphanol tartrate, and for butorphanol tartrate is 1.2, 1.0, and NLT 15 min, respectively.] Record a 30-min chromatogram. Preferably using an electronic integrator, determine the areas of all peaks in the chromatogram excluding the area of the solvent.
Butorphanol Tartrate
(Comment on this Monograph)id=m11000=Butorphanol Tartrate=B-Monos.pdf)
C21H29NO2 C4H6O6 477.55 Morphinan-3,14-diol, 17-(cyclobutylmethyl)-, ()-, [S(R*,R*)]-2,3-dihydroxybutanedioate (1:1) (salt); ()-17-(Cyclobutylmethyl)morphinan-3,14-diol D-()-tartrate (1:1) (salt) [58786-99-5]. DEFINITION Butorphanol Tartrate contains NLT 98.0% and NMT 102.0% of C21H29NO2 C4H6O6, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution, as obtained in Procedure 1. ASSAY PROCEDURE Sample solution: 500 mg of Butorphanol Tartrate in 75 mL of glacial acetic acid, and add crystal violet TS
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Butorphanol / Official Monographs

Calculate the percentage of synthesis precursors in the Sample solution taken: Result = (AV/AS) 100
USP 32
Standard solution: 5 mL of the Standard stock solution into a 50-mL volumetric flask containing 10.0 mL of Internal standard solution. Add water to volume, mix, and pass through a microporous filter, discarding the first 5 mL of the filtrate and collecting the remainder in a suitable container. Sample solution: Equivalent to 10 mg of butorphanol tartrate from Injection, into a 50-mL volumetric flask. Add 10.0 mL of Internal standard solution, mix, and add water to volume. Pass through a microporous filter, discarding the first 5 mL of the filtrate and collecting the remainder in a suitable container. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4-mm 30-cm; packing L11 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution (five replicate injections) [NOTEThe relative retention times for propylparaben and butorphanol tartrate are about 1.7 and 1.0, respectively.] Suitability requirements Relative standard deviation: NMT 1.5% Capacity factor: NLT 2.0 for butorphanol tartrate Analysis Samples: Standard solution and Sample solution Adjust the flow rate and other operating parameters, if necessary, until satisfactory chromatography and peak responses are obtained. Record the chromatograms, and measure the responses for the major peaks. Calculate the percentage of C21H29NO2 C4H6O6 in each mL of the Injection taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the butorphanol tartrate peak to the internal standard peak from the Sample solution = peak response ratio of the butorphanol tartrate RS peak to the internal standard peak from the Standard solution = concentration of USP Butorphanol Tartrate RS in CS the Standard solution (mg/mL) = nominal concentration of butorphanol tartrate in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS PH 791: 3.05.5 BACTERIAL ENDOTOXINS TEST 85: NMT 88.0 USP Endotoxin Units/mg of butorphanol tartrate OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Butorphanol Tartrate RS USP Endotoxin RS
= sum of the areas of all minor peaks AV = sum of the areas of the major and minor peaks AS Acceptance criteria: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 60 to 66 Sample solution: 4 mg/mL, in methanol WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Butorphanol Tartrate RS
Butorphanol Tartrate Injection

(Comment on this Monograph)id=m11010=Butorphanol Tartrate Injection=B-Monos.pdf) DEFINITION Butorphanol Tartrate Injection is a sterile solution of Butorphanol Tartrate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C21H29NO2 C4H6O6. It may contain a suitable preservative and a buffer. IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: USP Butorphanol Tartrate RS at the same concentration as the Injection Sample solution: Injection Chromatographic system (See Chromatography 621.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform, ethyl acetate, and methanol (40:10:9) Spray reagent: bromocresol purpl in dehydrated alcohol (1 in 250) Analysis Samples: Standard solution and Sample solution Develop the chromatogram until the solvent front has moved 10 cm above the line of application. Remove the plate, mark the solvent front, and allow the solvent to evaporate. Examine the plate under short-wavelength UV light. Visualize the butorphanol spots by lightly spraying the plate with Spray reagent: butorphanol appears as a blue spot against a light yellow background. Benzethonium chloride, if present, is observed as a streaked zone near the point of application. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.05 M ammonium acetate (1:3), adjusted by the addition of glacial acetic acid to a pH of 4.1 Internal standard solution: 0.2 mg/mL of propylparaben dissolved in methanol and diluted with water (1:49) Standard stock solution: 50 mg of USP Butorphanol Tartrate RS in a 25-mL volumetric flask containing 1.0 mL of 1 N sulfuric acid. Swirl the flask to dissolve the powder completely, add water to volume, and mix.
Butorphanol Tartrate Nasal Solution

(Comment on this Monograph)id=m11015=Butorphanol Tartrate Nasal Solution=B-Monos.pdf) DEFINITION Butorphanol Tartrate Nasal Solution is an aqueous solution of butorphanol tartrate for administration as a metered spray to
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the nasal mucosa. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of butorphanol tartrate (C21H29NO2 C4H6O6). IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 1.0 mg/mL of USP Butorphanol Tartrate RS in methanol Sample solution: Prepare a composite solution by pooling the contents of three containers of Nasal Solution into a suitable vessel. Transfer 1.0 mL of pooled sample to a 10-mL volumetric flask, and dilute with methanol to volume. Developing solvent system: Chloroform, methanol, benzene, and ammonium hydroxide (17:5:4:1) [CautionPrepare in a hood while wearing appropriate safety gloves, lab coat, and protective eyewear.] Spray reagent: Prepare a 1-in-10 solution of chloroplatinic acid in water. To 0.5 mL of this solution, add 33 mL of water and 1 g of potassium iodide. Prepare fresh daily. Analysis Samples: Standard solution and Sample solution Proceed as directed in the chapter, except to spray the plate with Spray reagent. Acceptance criteria: The typical RF value is 0.7 for butorphanol tartrate. ASSAY PROCEDURE Buffer: 3.4 g/L of monobasic potassium phosphate Mobile phase: Acetonitrile, triethylamine, and Buffer (15:2:85) Mix thoroughly, and adjust with 85.0% phosphoric acid to a pH of 3.0 0.1. Standard solution: 0.2 mg/mL of USP Butorphanol Tartrate RS in Mobile phase Mix, and filter, discarding the first 2 mL of the filtrate. [NOTEThe Standard solution is stable for at least 108 h.] Sample solution: Prepare a composite solution by pooling a minimum of four containers of Nasal Solution into a suitable glass vessel. Transfer the equivalent of 20 mg of butorphanol tartrate to a 100-mL volumetric flask. Dilute with Mobile phase to volume, mix, and filter, discarding the first 2 mL of the filtrate. [NOTEThe Sample solution has a nominal concentration of 0.2 mg/mL of butorphanol tartrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 15-cm; 5-m packing L11 Guard column: 4.6-mm 1-cm; 5-m packing L11 Temperature: 30 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H29NO2 C4H6O6 in the portion of Nasal Solution taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of butorphanol tartrate from the Sample solution rS
Official Monographs / Butorphanol 165

= peak response of butorphanol tartrate from the Standard solution CS = concentration of USP Butorphanol Tartrate RS in the Standard solution (mg/mL) = nominal concentration of butorphanol tatrtrate CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE Buffer: Prepare as directed in the Assay. Mobile phase: Acetonitrile, triethylamine, and Buffer (15:5.1:85) Mix thoroughly, and adjust with 85.0% phosphoric acid to a pH of 3.0 0.1. Standard solution: 0.005 mg/mL of USP Butorphanol Tartrate RS Sensitivity solution: Transfer 2.5 mL of the Standard solution to a 50-mL volumetric flask, dilute with water to volume, and mix. Do not filter. Sample solution: Prepare a composite solution by pooling a minimum of four containers of Nasal Solution into a suitable glass vessel. Transfer the equivalent of 50 mg of butorphanol tartrate to a 50-mL volumetric flask. Dilute with water to volume, and mix. Do not filter. [NOTEThe Sample solution has a nominal concentration of 1 mg/mL of butorphanol tartrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; 5-m packing L11 Guard column: 4.6-mm 1-cm; 5-m packing L11 Temperature: 40 Flow rate: 2.0 mL/min Injection size: 60 L System suitability Samples: Standard solution and Sensitivity solution Suitability requirements Relative standard deviation: NMT 10.0% in the Standard solution [NOTEThe peak height for butorphanol tartrate is greater than or equal to three times the baseline noise in the Sensitivity solution.] Analysis Samples: Standard solution and Sample solution Record the chromatograms, and measure the responses for the butorphanol tartrate peak in the Standard solution, and for all known and unknown related compounds in the Sample solution. The chromatographic run time is 40 min. Calculate the percentage of each related compound (see Impurity Table 1) and each unknown impurity in the portion of Nasal Solution taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response of each known or unknown related compound from the Sample solution = peak response of butorphanol tartrate from the Standard solution = concentration of USP Butorphanol Tartrate RS in the Standard solution (mg/mL) = nominal concentration of butorphanol tartrate in the Sample solution (mg/mL)
166
Butorphanol / Official Monographs

See Impurity Table 1
USP 32
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: The total aerobic microbial count does not exceed 1000 cfu/g or mL, and the total combined molds and yeasts count does not exceed 100 cfu/g or mL. It meets the requirements for absence of Staphylococcus aureus and Pseudomonas aeruginosa. PH 791: 4.06.0 OSMOLALITY AND OSMOLARITY 785: 252292 mOsmol/kg ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers at controlled room temperature. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Butorphanol Tartrate RS
Impurity Table 1 Relative Retention Time 0.3 0.7 1.0 Relative Response Factor Acceptance Criteria, NMT (%) 0.3 0.5 0.3 1.0
Name 3,14Dihydroxymorphinan 6-Butorphanol Butorphanol tartrate Unknown impurity Total
USP 32
Cabergoline Caffeine Caffeine Citrate Injection Caffeine Citrate Oral Solution Caffeine and Sodium Benzoate Injection Calamine Calamine Topical Suspension Phenolated Calamine Topical Suspension Calcifediol Calcifediol Capsules Calcitriol Calcitriol Injection Calcitonin Salmon Calcitonin Salmon Injection Calcitonin Salmon Nasal Solution Calcium Acetate Calcium Acetate Tablets Calcium Ascorbate Calcium Carbonate Calcium Carbonate Lozenges Calcium Carbonate Oral Suspension Calcium Carbonate Tablets Calcium Carbonate and Magnesia Tablets Calcium Carbonate and Magnesia Chewable Tablets Calcium Carbonate, Magnesia, and Simethicone Tablets Calcium Carbonate, Magnesia, and Simethicone Chewable Tablets Calcium and Magnesium Carbonates Oral Suspension Calcium and Magnesium Carbonates Tablets Calcium Chloride Calcium Chloride Injection Calcium Citrate Calcium Glubionate Syrup Calcium Gluceptate Calcium Gluceptate Injection Calcium Gluconate Calcium Gluconate Injection Calcium Gluconate Tablets Calcium Hydroxide Calcium Hydroxide Topical Solution Calcium Lactate Calcium Lactate Tablets Calcium Lactobionate Calcium Levulinate Calcium Levulinate Injection Calcium Pantothenate Calcium Pantothenate Tablets Racemic Calcium Pantothenate
MONOGRAPH LIST /
Dibasic Calcium Phosphate Dihydrate Anhydrous Dibasic Calcium Phosphate Dibasic Calcium Phosphate Tablets Calcium Polycarbophil Calcium Saccharate Calcium Undecylenate Camphor Camphor Spirit Capecitabine Capecitabine Tablets Capreomycin Sulfate Capreomycin for Injection Capsaicin Capsicum Capsicum Oleoresin Captopril Captopril Oral Solution Captopril Oral Suspension Captopril Tablets Captopril and Hydrochlorothiazide Tablets Carbachol Carbachol Intraocular Solution Carbachol Ophthalmic Solution Carbamazepine Carbamazepine Oral Suspension Carbamazepine Tablets Carbamazepine Extended-Release Tablets Carbamide Peroxide Carbamide Peroxide Topical Solution Carbenicillin Disodium Carbenicillin for Injection Carbenicillin Indanyl Sodium Carbenicillin Indanyl Sodium Tablets Carbidopa Carbidopa and Levodopa Tablets Carbinoxamine Maleate Carbinoxamine Maleate Tablets Carbol-Fuchsin Topical Solution
/ MONOGRAPH LIST
Cefadroxil for Oral Suspension Cefadroxil Tablets Cefamandole Nafate Cefamandole Nafate for Injection Cefazolin Cefazolin Sodium Cefazolin Injection Cefazolin for Injection Cefazolin Ophthalmic Solution Cefdinir Cefdinir Capsules Cefdinir for Oral Suspension Cefepime Hydrochloride Cefepime for Injection Cefixime Cefixime for Oral Suspension Cefixime Tablets Cefmenoxime Hydrochloride Cefmenoxime for Injection Cefmetazole Cefmetazole Injection Cefmetazole Sodium Cefmetazole for Injection Cefonicid Sodium Cefonicid for Injection Cefoperazone Sodium Cefoperazone Injection Cefoperazone for Injection Ceforanide Ceforanide for Injection Cefotaxime Sodium Cefotaxime Injection Cefotaxime for Injection Cefotetan Cefotetan Injection Cefotetan for Injection Cefotetan Disodium Cefotiam Hydrochloride Cefotiam for Injection Cefoxitin Sodium Cefoxitin Injection Cefoxitin for Injection Cefpiramide
USP 32
Carbon Dioxide Carbon Monoxide C 11 Flumazenil C 11 Injection Mespiperone C 11 Injection Methionine C 11 Injection Raclopride C 11 Injection Sodium Acetate C 11 Injection Urea C 13 Urea C 13 for Oral Solution Urea C 14 Capsules Carboplatin Carboplatin for Injection Carboprost Tromethamine Carboprost Tromethamine Injection Carboxymethylcellulose Sodium Carboxymethylcellulose Sodium Paste Carboxymethylcellulose Sodium Tablets Carisoprodol Carisoprodol Tablets Carisoprodol and Aspirin Tablets Carisoprodol, Aspirin, and Codeine Phosphate Tablets Carprofen Carprofen Tablets Carteolol Hydrochloride Carteolol Hydrochloride Ophthalmic Solution Carteolol Hydrochloride Tablets Casanthranol Cascara Sagrada Cascara Sagrada Extract Cascara Tablets Cascara Sagrada Fluidextract Aromatic Cascara Fluidextract Castor Oil Castor Oil Capsules Castor Oil Emulsion Aromatic Castor Oil Cefaclor Cefaclor Capsules Cefaclor for Oral Suspension Cefaclor Chewable Tablets Cefaclor Extended-Release Tablets Cefadroxil Cefadroxil Capsules
USP 32
Cefpiramide for Injection Cefpodoxime Proxetil Cefpodoxime Proxetil for Oral Suspension Cefpodoxime Proxetil Tablets Cefprozil Cefprozil for Oral Suspension Cefprozil Tablets Ceftazidime Ceftazidime Injection Ceftazidime for Injection Ceftizoxime Sodium Ceftizoxime Injection Ceftizoxime for Injection Ceftriaxone Sodium Ceftriaxone Injection Ceftriaxone for Injection Cefuroxime Axetil Cefuroxime Axetil for Oral Suspension Cefuroxime Axetil Tablets Cefuroxime Sodium Cefuroxime Injection Cefuroxime for Injection Oxidized Cellulose Oxidized Regenerated Cellulose Cellulose Sodium Phosphate Cellulose Sodium Phosphate for Oral Suspension Cephalexin Cephalexin Hydrochloride Cephalexin Capsules Cephalexin for Oral Suspension Cephalexin Tablets Cephalexin Tablets for Oral Suspension Cephalothin Sodium Cephalothin Injection Cephalothin for Injection Cephapirin Benzathine
MONOGRAPH LIST /
Cephapirin Benzathine Intramammary Infusion Cephapirin Sodium Cephapirin for Injection Cephapirin Sodium Intramammary Infusion Cephradine Cephradine Capsules Cephradine for Injection Cephradine for Oral Suspension Cephradine Tablets Cetylpyridinium Chloride Cetylpyridinium Chloride Lozenges Cetylpyridinium Chloride Topical Solution Activated Charcoal Chloral Hydrate Chloral Hydrate Capsules Chloral Hydrate Oral Solution Chlorambucil Chlorambucil Tablets Chloramphenicol Chloramphenicol Capsules Chloramphenicol Cream Chloramphenicol Injection Chloramphenicol Ophthalmic Ointment Chloramphenicol Ophthalmic Solution Chloramphenicol for Ophthalmic Solution Chloramphenicol Oral Solution Chloramphenicol Otic Solution Chloramphenicol Tablets Chloramphenicol and Hydrocortisone Acetate for Ophthalmic Suspension Chloramphenicol and Polymyxin B Sulfate Ophthalmic Ointment Chloramphenicol, Polymyxin B Sulfate, and Hydrocortisone Acetate Ophthalmic Ointment
USP 32
132270
Official Monographs / Cabergoline 1

Add the following:

v
Cabergoline
(Comment on this Monograph)id=m11140=Cabergoline=CaChl-Monos.pdf)
C26H37N5O2 451.60 Ergoline-8-carboxamide, N-[3-(dimethylamino)propyl]-N[(ethy- quinolamino)carbonyl]-6-(2-propenyl)-; 1-[(6-Allylergolin-8-yl)carbonyl]-1-[3-(dimethylamino)propyl]-3ethylurea [81409-90-7]. DEFINITION Cabergoline contains NLT 98.0% and NMT 102.0% of the labeled amount of C26H37N5O2, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTE Prepare solutions immediately before use and protect from light.] Buffer: Dissolve 6.8 g of monobasic potassium phosphate in 900 mL water, adjust with phosphoric acid to a pH of 2.0, and dilute to 1 L. Add 0.2 mL of triethylamine to the resulting solution, and mix. Mobile phase: Acetonitrile and Buffer (4:21) Standard solution: 0.25 mg/mL of USP Cabergoline RS in Mobile phase [NOTESonicate if needed.] Sample solution: 0.25 mg/mL of Cabergoline in Mobile phase [NOTESonicate if needed.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 25-cm; 10 m packing L1 Flow rate: 1.3 mL/min Injection size: 100 L System suitability Samples: Standard solution Suitability requirements Column efficiency: NLT 1000 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C26H37N5O2 in the portion of Cabergoline taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cabergoline RS in the Standard solution (mg/mL) = concentration of caberrgoline the Sample solution (mg/mL)
IMPURITIES Organic Impurities PROCEDURE [NOTEPrepare solutions immediately before use, and protect from light.] Buffer: Proceed as directed in the Assay. Mobile phase: Proceed as directed in the Assay. System suitability solution: To 10 mL of 0.1 M sodium hydroxide, add 50 mg of Cabergoline. Stir for about 15 min. To 1 mL of the suspension, add 1 mL of 0.1 M hydrochloric acid, and dilute with Mobile phase to 10.0 mL. Sonicate until dissolution is complete. [NOTEThe main degradation product obtained is cabergoline related compound A.] Sample solution: 0.25 mg/mL of Cabergoline in Mobile phase [NOTESonicate if needed.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.0-mm 25-cm; 10 m packing L1 Flow rate: 1.3 mL/min Injection size: 100 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 3.0 between cabergoline and cabergoline related compound A Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Cabergoline taken: Result = (rU/rS) 100 = peak response of each impurity from the Sample solution = sum of the peak responses for all peaks from the rS Sample solution Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 0.8% rU
Impurity Table 1 Relative Retention Time 0.3 0.6 0.8 1.0 Acceptance Criteria, NMT (%) 0.1 0.1 0.3
Name Cabergoline related compound Da Cabergoline related compound Bb Cabergoline related compound Ac Cabergoline
a
(6aR,9R,10aR)-N-[3-(Dimethylamino)propyl]-7-(prop-2enyl)-4,6,6a,7,8,9,10,10aoctahydroindolo[4,3-fg]quinoline-9-carboxamide. b (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-7-(prop-2enyl)6a,7,8,9,10,10a-hexahydroindolo[4,3-fg]quinoline-4,9(6H)dicarboxamide. c (6aR,9R,10aR)-7-(Prop-2-enyl)-4,6,6a,7,8,9,10,10a-octahydroindolo[4,3fg]quinoline-9-carboxylic acid. d (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-N9(ethylcarbamoyl)-7-(prop2-enyl)-6a,7,8,9,10,10a-hexahydroindolo[4,3fg]quinoline-4,9(6H)-dicarboxamide.
Cabergoline / Official Monographs

Impurity Table 1 (continued) Relative Retention Time 2.9 Acceptance Criteria, NMT (%) 0.3 0.10
USP 32
ASSAY PROCEDURE Solution A: 0.82 mg/mL of anhydrous sodium acetate Mobile phase: Acetonitrile, tetrahydrofuran, and Solution A (5:4:191). Adjust with glacial acetic acid to a pH of 4.5. System suitability solution: 0.02 mg/mL of theophylline in Mobile phase [NOTEShake, and sonicate, if necessary, to dissolve.] Standard solution: 0.2 mg/mL of USP Caffeine RS in System suitability solution and Mobile phase (1:4) [NOTE Shake, and sonicate, if necessary, to dissolve.] Sample solution: 0.2 mg/mL of Caffeine in Mobile phase [NOTEShake, and sonicate, if necessary, to dissolve.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for theophylline and caffeine are about 0.69 and 1.0, respectively. ] Suitability requirements Resolution: NLT 6.0 between theophylline and caffeine Tailing factor: NMT 2.0 for each of the peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H10N4O2 in the portion of Caffeine taken: Result = (rU/rS) (CS/CU) 100
Name Cabergoline related compound Cd Any other individual, unidentified impurity

a
(6aR,9R,10aR)-N-[3-(Dimethylamino)propyl]-7-(prop-2enyl)-4,6,6a,7,8,9,10,10aoctahydroindolo[4,3-fg]quinoline-9-carboxamide. b (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-7-(prop-2enyl)6a,7,8,9,10,10a-hexahydroindolo[4,3-fg]quinoline-4,9(6H)dicarboxamide. c (6aR,9R,10aR)-7-(Prop-2-enyl)-4,6,6a,7,8,9,10,10a-octahydroindolo[4,3fg]quinoline-9-carboxylic acid. d (6aR,9R,10aR)-N9-[3-(Dimethylamino)propyl]-N4-ethyl-N9(ethylcarbamoyl)-7-(prop2-enyl)-6a,7,8,9,10,10a-hexahydroindolo[4,3fg]quinoline-4,9(6H)-dicarboxamide.
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 77 to 83 Sample solution: 1 mg/mL in alcohol, on the anhydrous basis WATER DETERMINATION, Method I 921: NMT 5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. USP REFERENCE STANDARDS 11 USP Cabergoline RSvUSP32
Caffeine
(Comment on this Monograph)id=m11170=Caffeine=Ca-ChlMonos.pdf)
= peak response for Caffeine from the Sample solution = peak response for Caffeine from the Standard rS solution = concentration of USP Caffeine RS in the Standard CS solution (mg/mL) = nominal concentration of caffeine in the Sample CU solution (mg/mL) Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE 1: READILY CARBONIZABLE SUBSTANCES 271: Dissolve 0.5 g in 5 mL of sulfuric acid TS: the solution has no more color than Matching Fluid D. PROCEDURE 2 Mobile phase, System suitability solution, Standard solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Caffeine taken: Result = (ri/rT) 100 ri rT = peak response for each impurity, Sample solution = sum of the responses of all the peaks, Sample solution
rU
C8H10N4O2 (anhydrous) 1H-Purine-2,6-dione, 3,7-dihydro-1,3,7-trimethyl-; 1,3,7-Trimethylxanthine. [58-08-2] Monohydrate [5743-12-4].
194.19 212.21
DEFINITION Caffeine is anhydrous or contains one molecule of water of hydration. It contains NLT 98.5% and NMT 101.0% of C8H10N4O2, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. PROCEDURE Sample solution: 5 mg in 1 mL of hydrochloric acid, in a porcelain dish Analysis: Add 50 mg of potassium chlorate, and evaporate on a steam bath to dryness. Invert the dish over a vessel containing a few drops of 6 N ammonium hydroxide. Acceptance criteria: The residue acquires a purple color, which disappears upon the addition of a solution of 1 N sodium hydroxide.
USP 32
Acceptance criteria Individual impurities: NMT 0.1% Total impurities: NMT 0.1% SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 235239, determined after drying at 80 for 4 h WATER DETERMINATION, Method III 921: Dry it at 80 for 4 h: the anhydrous form loses NMT 0.5% and the hydrous form NMT 8.5% of its weight. OTHER ALKALOIDS: To 5 mL of a solution (1 in 50), add mercuricpotassium iodide TS: no precipitate is formed. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve hydrous Caffeine in tight containers. Preserve anhydrous Caffeine in well-closed containers. LABELING: Label it to indicate whether it is anhydrous or hydrous. USP REFERENCE STANDARDS 11 USP Caffeine RS
Official Monographs / Caffeine 3

Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 150-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for theophylline and caffeine are 0.7 and 1.0, respectively. ] Suitability requirements Resolution: NLT 6.0 between theophylline and caffeine Tailing factor: NMT 2.0, theophylline and caffeine Relative standard deviation: NMT 2.0% for caffeine Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H18N4O9 in the volume of Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Caffeine RS in the Standard solution (mg/mL) = nominal concentration of caffeine citrate in the CU Sample solution (mg/mL) = molecular weight of caffeine citrate, 386.31 Mr1 = molecular weight of caffeine, 194.19 Mr2 Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE Mobile phase, Identification solution, Standard solution, and Sample solution: Proceed as directed in the Assay. System suitability solution: Standard solution and water (1:39) Chromatographic system: Proceed as directed in the Assay. Suitability requirements Sensitivity: A peak for theophylline is discernable. Analysis Samples: Standard solution and Sample solution Calculate the percentage of any impurity in the portion of Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) F 100 rU rS CS CU Mr1 Mr2 F = individual peak response for each impurity in the Sample solution = peak response for caffeine from the Standard solution = concentration of USP Caffeine RS in the Standard solution (mg/mL) = nominal concentration of caffeine citrate in the Sample solution (mg/mL) = molecular weight of caffeine citrate, 386.31 = molecular weight of caffeine, 194.19 = relative response factor as given in Impurity Table 1 rU rS CS
Caffeine Citrate Injection

(Comment on this Monograph)id=m11175=Caffeine Citrate Injection=Ca-Chl-Monos.pdf) DEFINITION Caffeine Citrate Injection is a sterile solution containing Caffeine and citric acid in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of caffeine citrate (C14H18N4O9). It contains no bacteriostat or other preservative. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Citrate 191 C. PROCEDURE Sample solution: 4 g of potassium iodide in a 100-mL volumetric flask. Add 10 mL of water, and shake until the potassium iodide is dissolved. Transfer 2 g of iodine to the volumetric flask, and shake until dissolved. Dilute with water to volume. Transfer 5 drops to a 25-mL centrifuge tube containing 5.0 mL of Injection, and mix. Analysis: Add 0.5 mL of 2.0 M hydrochloric acid solution, and mix. Acceptance criteria: A brown precipitate that dissolves on neutralization with 0.5 mL of sodium hydroxide TS is produced. ASSAY PROCEDURE Mobile phase: Acetonitrile, tetrahydrofuran, and 0.01 M sodium acetate (5:4:191). Adjust with glacial acetic acid to a pH of 4.5. Identification solution: 0.02 mg/mL of theophylline Standard solution: 0.2 mg/mL of USP Caffeine RS in Identification solution and water (1:4) Sample solution: Equivalent to 0.2 mg/mL of caffeine from a volume of Injection in water. [NOTEPass through a polyvinylidene difluoride or equivalent membrane having a porosity of 0.45 m.]
Caffeine / Official Monographs

Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 0.1%
Impurity Table 1 Relative Retention Time 0.4 0.6 0.7 Relative Response Factor 0.878 1.10 0.905 1.0 Acceptance Criteria, NMT (%) 0.1 0.1 0.1 0.1
USP 32
ASSAY PROCEDURE Mobile phase: Acetonitrile, tetrahydrofuran, and 0.01 M sodium acetate (5:4:191). Adjust with glacial acetic acid to a pH of 4.5. Identification solution: 0.02 mg/mL of theophylline Standard solution: 0.2 mg/mL of USP Caffeine RS in Identification solution and water (1:4) Sample solution: Equivalent to 0.2 mg/mL of caffeine from a volume of Oral Solution in water. [NOTEPass through a polyvinylidene difluoride or equivalent membrane having a 0.45-m porosity.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 150-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for theophylline and caffeine are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 6.0 between theophylline and caffeine Tailing factor: NMT 2.0 determined from the theophylline and caffeine peaks Relative standard deviation: NMT 2.0% for caffeine peaks Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H18N4O9 in the volume of Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Caffeine RS in the Standard solution (mg/mL) = nominal concentration of caffeine citrate in the CU Sample solution (mg/mL) Mr1 = molecular weight of caffeine citrate, 386.31 = molecular weight of caffeine, 194.19 Mr2 Acceptance criteria: 90.0%110.0% rU rS CS IMPURITIES Organic Impurities PROCEDURE Mobile phase, Identification solution, Standard solution, and Sample solution: Proceed as directed in the Assay. System sensitivity solution: Standard solution and water (1:39) Chromatographic system: Proceed as directed in the Assay. C Suitability requirements Sensitivity: A peak for theophylline is discernable. Analysis Samples: Standard solution and Sample solution Calculate the percentage of any impurity in the portion of Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS = individual peak response for each impurity for the Sample solution = peak response for caffeine from the Standard solution
Name Theobromine Paraxanthine Theophylline Any other impurity
SPECIFIC TESTS COLOR AND CLARITY: Transfer a suitable portion of the Injection to a clear glass test tube, and visually examine the solution in a well-lighted area: the solution is colorless and free of haze, obvious turbidity, and precipitate. BACTERIAL ENDOTOXINS TEST 85: NMT 0.25 USP Endotoxin Unit/mg of caffeine STERILITY TESTS 71: It meets the requirements for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 4.25.2 PARTICULATE MATTER IN INJECTIONS 788: NMT 150 particles are equal to or greater than 10 m, and NMT 25 particles are equal to or greater than 25 m OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose, tight containers of Type I glass, and store between 15 and 30. USP REFERENCE STANDARDS 11 USP Caffeine RS USP Endotoxin RS
Caffeine Citrate Oral Solution

(Comment on this Monograph)id=m11180=Caffeine Citrate Oral Solution=Ca-Chl-Monos.pdf) DEFINITION Caffeine Citrate Oral Solution is a sterile aqueous solution containing Caffeine and citric acid. It contains NLT 90.0% and NMT 110.0% of the labeled amount of caffeine citrate (C14H18N4O9). It contains no bacteriostat or other preservative. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFCATION TESTSGENERAL, Citrate 191 C. PROCEDURE Sample solution: 10 mg/mL of potassium iodide and 20 mg/ml of iodine in water [NOTEInitially combine potassium iodide and water, and shake until dissolved, then add iodine to the solution] Analysis: Transfer 5 drops of the Sample solution to a 25mL centrifuge tube containing 5 mL of the Oral Solution, and mix. Add 0.5 mL of 2.0 M hydrochloric acid solution, and mix. Acceptance criteria: A brown precipitate is produced that dissolves on neutralization with 0.5 mL of sodium hydroxide TS.
USP 32
= concentration of USP Caffeine RS in the Standard solution (mg/mL) = nominal concentration of caffeine citrate in the CU Sample solution (mg/mL) = molecular weight of caffeine citrate, 396.31 Mr1 = molecular weight of caffeine, 194.19 Mr2 Acceptance criteria Individual impurities: See Impurity Table I. Total impurities: NMT 0.1%
Official Monographs / Calamine 5

caffeine, passing each chloroform extract through a small filter previously moistened with chloroform into a tared dish. [NOTERetain the water layer for the Assay for Sodium Benzoate.] Wash the stem of the separator, the filter, and the funnel with 10 mL of hot chloroform, adding the washings to the dish. Evaporate the combined chloroform solutions on a steam bath, adding 2 mL of alcohol just before the last trace of chloroform is expelled. Complete the evaporation of the solvent; dry the residue, consisting of C8H10N4O2, at 80 for 4 h; cool; and weigh. Acceptance criteria: 90.0%110.0% SODIUM BENZOATE Sample solution: To the water layer obtained in the Assay for Caffeine add 75 mL of ether and 5 drops of methyl orange TS. Analysis: Titrate with 0.1 N hydrochloric acid VS, mixing the liquids by vigorous shaking, until a permanent pink color is produced in the water layer. Each mL of 0.1 N hydrochloric acid is equivalent to 14.41 mg of C7H5NaO2. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.7 USP Endotoxin Unit/mg of caffeine and sodium benzoate, based on the total, in mg, of the labeled amounts PH 791: 6.58.5 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Caffeine RS USP Endotoxin RS
CS
Name Theobromine Paraxanthine Theophylline Any other individual, unidentified impurity
SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 4.25.2 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose, tight containers, and store at a temperature between 15 and 30. USP REFERENCE STANDARDS 11 USP Caffeine RS
Caffeine and Sodium Benzoate Injection

(Comment on this Monograph)id=m11200=Caffeine and Sodium Benzoate Injection=Ca-Chl-Monos.pdf) DEFINITION Caffeine and Sodium Benzoate Injection is a sterile solution containing equal amounts of Caffeine and Sodium Benzoate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amounts of anhydrous caffeine (C8H10N4O2) and sodium benzoate (C7H5NaO2). IDENTIFICATION A. INFRARED ABSORPTION 197K: The caffeine obtained in the Assay for Caffeine meets the requirements of the test. B. Dip the end of a platinum wire into a portion of Injection, and introduce it into a nonluminous flame: the flame is colored intensely yellow. C. To 0.5 mL of Injection, add a few drops of ferric chloride TS: a salmon-colored precipitate is formed. To another portion of Injection, add 3 N hydrochloric acid: a white precipitate is formed. ASSAY CAFFEINE Sample solution: Equivalent to 250 mg each of caffeine and sodium benzoate from a volume of Injection. Transfer it completely with the aid of 5 mL of water to a small separator, add 1 drop of phenolphthalein TS, and add 0.1 N sodium hydroxide, dropwise, until a permanent pink color is just produced. Analysis: Shake the mixture with three or more 20-mL portions of chloroform to effect complete extraction of the
Calamine
(Comment on this Monograph)id=m11250=Calamine=Ca-ChlMonos.pdf) Iron oxide (Fe2O3), mixture with zinc oxide; Calamine (pharmaceutical solution) [8011-96-9]. DEFINITION Calamine is Zinc Oxide with a small proportion of ferric oxide, and contains, after ignition, NLT 98.0% and NMT 100.5% of ZnO. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Zinc 191: Combine 1 g of Calamine with 10 mL of 3 N hydrochloric acid, and filter: the filtrate meets the requirements of tests. B. PROCEDURE Analysis: Combine 1g of Calamine with 100 mL of 3 N hydrochloric acid, heat to boiling, and filter. Acceptance criteria: The filtrate assumes a reddish color upon the addition of ammonium thiocyanate TS. ASSAY PROCEDURE Sample solution: Digest 1.5 g of freshly ignited Calamine in 50.0 mL of 1 N sulfuric acid VS, applying gentle heat until no further solution occurs. Filter the mixture, and wash the residue on the filter with hot water until the last washing is neutral to litmus paper. To the combined filtrate and washings, add 2.5 g of ammonium chloride, cool, add methyl orange TS. Analysis: Titrate with 1 N sodium hydroxide VS. Each mL of 1 N sulfuric acid is equivalent to 40.69 mg of ZnO.
Calamine / Official Monographs

USP 32
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
IMPURITIES Inorganic Impurities LOSS ON IGNITION 733: Weigh 2 g, and ignite at 500 to constant weight: it loses NMT 2.0% of its weight. CALCIUM: Digest 1 g in 25 mL of 3 N hydrochloric acid for 30 min, filter to remove the insoluble ferric oxide, and add 6 N ammonium hydroxide to the filtrate until the precipitate first formed is redissolved, then add 5 mL more of 6 N ammonium hydroxide. To 10 mL of this solution, add 2 mL of ammonium oxalate TS: NMT a slight turbidity is produced. CALCIUM OR MAGNESIUM: To another 10-mL portion of the solution prepared in the Calcium Procedure, add 2 mL of dibasic sodium phosphate TS: NMT a slight turbidity is produced. ARSENIC, Method I 211: NMT 8 ppm LEAD: To 1 g add 15 mL of water, stir, then add 3 mL of glacial acetic acid, and warm on a steam bath until dissolved. Filter, and add 5 drops of potassium chromate TS: no turbidity is produced. SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. ACID-INSOLUBLE SUBSTANCES: Dissolve 2.0 g in 50 mL of 3 N hydrochloric acid. If an insoluble residue remains, collect it on a tared filter, wash with water, dry at 105 for 1 h, cool, and weigh: the weight of the residue does not exceed 40 mg (2.0%). ALKALINE SUBSTANCES: Digest 1.0 g with 20 mL of water on a steam bath for 15 min, filter, and add 2 drops of phenolphthalein TS: if a red color is produced, NMT 0.20 mL of 0.10 N sulfuric acid is required to discharge it. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Phenolated Calamine Topical Suspension

(Comment on this Monograph)id=m11270=Phenolated Calamine Topical Suspension=Ca-Chl-Monos.pdf) Prepare Phenolated Calamine Topical Suspension as follows:
Liquefied Phenol Calamine Topical Suspension To make 10 mL 990 mL 1000 mL
Mix the ingredients. [NOTEShake Phenolated Calamine Topical Suspension before dispensing.] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Calcifediol
(Comment on this Monograph)id=m11290=Calcifediol=Ca-ChlMonos.pdf)
Calamine Topical Suspension

(Comment on this Monograph)id=m11260=Calamine Topical Suspension=Ca-Chl-Monos.pdf) (Former monograph title is Calamine Lotion) Prepare Calamine Topical Suspension as follows.
Calamine Zinc Oxide Glycerin Bentonite Magma Calcium Hydroxide Topical Solution To make 80 g 80 g 20 mL 250 mL A sufficient quantity 1000 mL
C27H44O2 H2O 418.65 9,10-Secocholesta-5,7,10(19)-triene-3,25-diol monohydrate, (3,5Z,7E)-; 25-Hydroxycholecalciferol; Monohydrate [63283-36-3]. DEFINITION Calcifediol contains NLT 97.0% and NMT 103.0% of C27H44O2 H2O. IDENTIFICATION A. INFRARED ABSORPTION 197M ASSAY PROCEDURE Internal standard solution: 0.10 mg/mL of testosterone in ethyl acetate Mobile phase: Heptane, methylene chloride, ethyl acetate, and water-saturated heptane (6:3:5:6) Standard solution: 20 g/mL of USP Calcifediol RS in Internal standard solution Sample solution: 20 g/mL of Calcifediol in Internal standard solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm, and a pump capable of providing constant flow at over 2000 psi
Dilute the Bentonite Magma with an equal volume of Calcium Hydroxide Topical Solution. Mix the powders intimately with the Glycerin and about 100 mL of the diluted magma, triturating until a smooth, uniform paste is formed. Gradually incorporate the remainder of the diluted magma. Finally, add enough Calcium Hydroxide Topical Solution to make 1000 mL, and shake. If a more viscous consistency in the Calamine Topical Suspension is desired, the quantity of Bentonite Magma may be increased to NMT 400 mL. [NOTEShake the Calamine Topical Suspension before dispensing.]
USP 32
Column: 4-mm 30-cm; packing L3 Injection size: Equal volumes System suitability Sample: Standard solution Suitability requirements Resolution factor: NLT 3.0 Relative standard deviation: NMT 3.0% (four replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percent of C27H44O2 H2O taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of calcifediol to testosterone from the Sample solution = peak response ratio of calcifediol to testosterone RS from the Standard solution = concentration of USP Calcifediol RS in the CS Standard solution (g/mL) = concentration of Calcifediol in the Sample CU solution (g/mL) Acceptance criteria: 97.0%103.0% RU SPECIFIC TESTS WATER DETERMINATION, Method Ia 921: determined on a 0.2 g specimen 3.8%5.0%,
Official Monographs / Calcifediol 7

Mobile phase: Heptane, methylene chloride, ethyl acetate, and water-saturated heptane (6:3:5:6) Standard solution: 7 g/mL of USP Calcifediol RS in Internal standard solution Sample solution: 7 g/mL of calcifediol from Capsules in Internal standard solution [NOTEUsing a suitable implement, shear open a number of Capsules inside the container. Wash the implement with a volume of Internal standard solution that will yield the final concentration. Collect the rinsings in the container, and mix to obtain a homogeneous solution of the Capsule contents.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm and a pump capable of providing constant flow of greater than 2000 psi Column: 4-mm 30-cm; packing L3 Injection size: Equal volumes System suitability Sample: Standard solution Suitability requirements Resolution factor: NLT 3.0 Relative standard deviation: NMT 3.0% (four replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percent label claim of C27H44O2 H2O taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of calcifediol to testosterone of the Sample solution = peak response ratio of calcifediol to testosterone RS of the Standard solution CS = concentration of USP Calcifediol RS in the Standard solution (g/mL) = concentration of calcifediol in the Sample CU solution (g/mL) Acceptance criteria: 90.0%120.0% RU PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 500 mL Apparatus 2: 50 rpm Time: 15 min Analysis: Place 1 Capsule in each vessel, and allow the Capsule to sink to the bottom of the vessel before starting rotation of the blade. Observe the Capsules, and record the time taken for each capsule shell to rupture. Tolerances: Meets the requirements if all of the Capsules tested rupture in NMT 15 min. If 1 or 2 of the Capsules rupture in more than 15 but NMT 30 min, repeat the test on 12 additional Capsules. NMT 2 of the total of 18 Capsules tested rupture in more than 15 but NMT 30 min. UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Calcifediol RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers at controlled room temperature. USP REFERENCE STANDARDS 11 USP Calcifediol RS
Calcifediol Capsules
(Comment on this Monograph)id=m11293=Calcifediol Capsules=Ca-Chl-Monos.pdf) DEFINITION Calcifediol Capsules contain NLT 90.0% and NMT 120.0% of the labeled amount of C27H44O2 H2O. IDENTIFICATION PROCEDURE Standard solution: 150 g/mL of USP Calcifediol RS in chloroform Sample solution: Equivalent to 150 g of calcifediol from Capsules, in 1 mL of methanol, and shake vigorously for 1 min. Separate the layers by centrifugation, and transfer as much of the top, methanol layer as possible to a second container. Evaporate this extract to dryness, and dissolve the residue in 1 mL of chloroform. Analysis: Proceed as directed under Thin-Layer Chromatographic Identification Test 201. Application volume: 20 L Developing solvent system: Cyclohexane and ethyl acetate (3:2) ASSAY PROCEDURE Internal standard solution: ethyl acetate
35 g/mL of testosterone in
Calcitriol / Official Monographs
USP 32
Column efficiency: NLT 10,000 theoretical plates Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C27H44O3 in the portion of Calcitriol taken: Result = (rU/rS) (CS/CU) 100 = sum of the peak responses of calcitriol and precalcitriol from the Sample solution rS = sum of the peak responses of calcitriol and precalcitriol from the Standard solution CS = concentration of USP Calcitriol RS in the Standard solution (g/mL) CU = concentration of Calcitriol in the Sample solution (g/mL) Acceptance criteria: 97.0%103.0% (anhydrous form, calculated on the solvent-free basis); 97.0%103.0% (monohydrate form, calculated on the anhydrous basis) IMPURITIES Organic Impurities PROCEDURE [NOTECarry out the procedure as rapidly as possible, avoiding unnecessary exposure of solutions to light and air.] Solution A, Mobile phase, System suitability solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay. Analysis Sample: Sample solution [NOTERecord chromatograms for at least two times the retention time of the calcitriol peak.] Calculate the percentage of any individual impurity in the portion of Calcitriol taken: Result = (ri/rs) 100 = peak response of any individual peak other than the main calcitriol peak and the pre-calcitriol peak = sum of the responses of all the peaks rs Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 1.0% [NOTEDisregard any peak less than 0.1%.] ri
Impurity Table 1 Relative Retention Time 0.43 0.96 1.15 1.5 Relative Response Factor Acceptance Criteria, NMT (%) 0.1 0.25 0.1 0.25 0.1
Calcitriol
(Comment on this Monograph)id=m11360=Calcitriol=Ca-ChlMonos.pdf)
rU
C27 H44 O3 416.64 9,10-Secocholesta-5,7,10(19)-triene-1,3,25-triol, (1,3,5Z,7E)-; (5Z,7E)-9,10-Secocholesta-5,7,10(19)-triene-1,3,25-triol [32222-06-3]. Monohydrate 434.65 [77326-95-5]. DEFINITION Calcitriol is anhydrous or contains one molecule of hydration. The anhydrous form contains NLT 97.0% and NMT 103.0% of C27 H44 O3, calculated on the solvent-free basis. The monohydrate form contains NLT 97.0% and NMT 103.0% of C27H44O3, calculated on the anhydrous basis. [CAUTIONCare should be taken to prevent inhaling particles of calcitriol, and exposing the skin to it.] IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTECarry out the procedure as rapidly as possible, avoiding unnecessary exposure of solutions to light and air.] Solution A: 1.0 mg/mL of tris(hydroxymethyl)aminomethane in water, adjusted with phosphoric acid to a pH of 7.07.5 Mobile phase: Acetonitrile and Solution A (11:9) Standard solution: 100 g/mL of calcitriol from USP Calcitriol RS, dissolve first in acetonitrile (without heating), using 55% of the final volume, then dilute with Solution A to volume [NOTEAllow the solution to warm up to room temperature before diluting with Solution A to final volume.] System suitability solution: Heat 2.0 mL of the Standard solution at 80 for 30 min. Sample solution: 100 g/mL of Calcitriol, dissolve first in acetonitrile (without heating), using 55% of the final volume, then dilute with Solution A to volume [NOTEAllow the solution to warm up to room temperature before diluting with Solution A to final volume.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; 5-m packing L7 Column temperature: 40 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: Standard solution and System suitability solution. [NOTEThe relative retention times for pre-calcitriol and calcitriol are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.5 between the pre-calcitriol and calcitriol peaks
Name Triazoline adduct of pre-calcitriol trans-Calcitriola 1-Calcitriolb Methylene calcitriolc Any other individual unidentified impurity
a b
(5E,7E)-9,10-secocholesta-5,7,10(19)-triene-1,3,25-triol (5Z,7E)-9,10-secocholesta-5,7,10(19)-triene-1,3,25-triol c(5Z,7E)-1,3-dihydroxy-17-((R)-7-hydroxy-7-methyloctan-2-yl)-9,10secoandrosta-5,7,10(19)-triene
SPECIFIC TESTS WATER DETERMINATION, Method Ic 921: as a monohydrate, 3.5%5.5%
Where it is labeled
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store as per labeling instructions. LABELING: Where it is a monohydrate form, the label so indicates. USP REFERENCE STANDARDS 11 USP Calcitriol RS
Official Monographs / Calcitonin 9

PH 791: 5.98.0, determined on a portion to which, if necessary, 0.30 mL of saturated potassium chloride solution has been added for each 100 mL of Injection OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass, protected from light. Store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Calcitriol Solution RS USP Endotoxin RS
Calcitriol Injection
(Comment on this Monograph)id=m11380=Calcitriol Injection=Ca-Chl-Monos.pdf) DEFINITION Calcitriol Injection is a sterile solution of Calcitriol. It contains an amount of Calcitriol equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of calcitriol (C27H44O3). It contains no antimicrobial agents. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEAvoid unnecessary exposure of solutions to light or air.] Mobile phase: Methanol and water (37:13). Make adjustments if necessary so that the retention time of calcitriol is NLT 20 min. Standard solution: 3.0 mL of USP Calcitriol Solution RS, equilibrated to room temperature, to a container, and add 3.0 mL of water Sample solution: Equivalent to 3 g of calcitriol from a volume of Injection, in a sufficient amount of water to dilute to a total volume of 3.0 mL, and add 3.0 mL of methanol Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 264 nm Column: 4.6-mm 7.5-cm; 3-m packing L1 Guard column: 4.6-mm 4.5-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C27H44O3 in the portion of Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of calcitriol in the USP Calcitriol Solution RS (g/mL) = nominal concentration of calcitriol in the Sample CU solution (g/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 100 USP Endotoxin Units/g of calcitriol PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections rU rS CS
Calcitonin Salmon
(Comment on this Monograph)id=m11340=Calcitonin Salmon=Ca-Chl-Monos.pdf)
C145H240N44O48S2 [47931-85-1].
3432 daltons
DEFINITION Calcitonin Salmon is a polypeptide that has the same sequence as that of the hormone that regulates calcium metabolism and is secreted by the ultimobranchial gland of salmon. It is produced from either synthetic processes or microbial processes using recombinant DNA (rDNA) technology. The host cell-derived protein content and the host cell- or vectorderived DNA content of Calcitonin Salmon produced from an rDNA process are determined by validated methods. It contains NLT 90.0% and NMT 105.0% of calcitonin salmon, calculated on an acetic acid-free and dried basis. [NOTEOne mg of acetic acid-free, anhydrous Calcitonin Salmon is equivalent to 6000 USP Calcitonin Salmon Units.] IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay ASSAY PROCEDURE Solution A: Dissolve 3.26 g of tetramethylammonium hydroxide pentahydrate in 900 mL of water, add 100 mL of acetonitrile, and mix. Adjust with phosphoric acid to a pH of 2.5. Solution B: Dissolve 1.45 g of tetramethylammonium hydroxide pentahydrate in 400 mL of water, add 600 mL of acetonitrile, and mix. Adjust with phosphoric acid to a pH of 2.5. Mobile phase: See gradient table below.
10
Calcitonin / Official Monographs

rU
USP 32
= peak area response of each impurity in the Sample solution = sum of all areas in the Sample solution rS Acceptance criteria: Individual impurities: NMT 3.0% Total impurities: NMT 5.0% PROCEDURE 2 [NOTEThis test needs to be performed only on material produced using rDNA technology.] Buffer A: 2.72 mg/mL of monobasic potassium phosphate in water Buffer B: 2.72 mg/mL of monobasic potassium phosphate and 29.2 mg/mL of sodium chloride in water Buffer C: 4.8 mg/mL of citric acid in water. Adjust with 1 M sodium hydroxide to a pH of 3.0, prior to final dilution Solution A: Acetonitrile and Buffer A (3:17). Adjust with 45% w/w potassium hydroxide to a pH of 5.0 Solution B: Acetonitrile and Buffer B (3:17). Adjust with 45% w/w potassium hydroxide to a pH of 4.6 Mobile phase: See gradient table below.
Time (min) 0 10 15 15.1 22.1 Solution A (%) 100 0 0 100 100 Solution B (%) 0 100 100 0 0
Standard solution: 1.0 mg of USP Calcitonin Salmon RS in Solution A System suitability solution: Dissolve the contents of a vial of USP Calcitonin Salmon Related Compound A RS in 0.4 mL of Solution A, add 0.1 mL of the Standard solution, and mix. Sample solution: 1.0 mg/mL of Calcitonin Salmon in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Column temperature: 65 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for calcitonin salmon and calcitonin salmon related compound A are 1.0 and 1.15, respectively.] Suitability requirements Resolution: NLT 3 between calcitonin salmon and calcitonin salmon related compound A Tailing factor: NMT 2.5 for calcitonin salmon Relative standard deviation: NMT 3% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C145H240N44O48S2 in the portion of Calcitonin Salmon taken: Result = (rU/rS) (CS/CU) 100 = peak response of Calcitonin Salmon from the Sample solution = peak response of calcitonin salmon from the rS Standard solution = concentration of USP Calcitonin Salmon RS in CS the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 90.0%105.0% rU OTHER COMPONENTS ACETIC ACID CONTENT 503 Sample solution: 1 mg/mL of Calcitonin Salmon in Diluent (see Acetic acid in peptides, Diluent) Acceptance criteria: 4%20% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 50 ppm Organic Impurities PROCEDURE 1 [NOTEThis test is performed on material produced by both chemical processes and recombinant DNA processes.] Solution A, Solution B, Mobile phase, System suitability solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Sample solution: Prepare as directed for the Sample solution under Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Calcitonin Salmon taken: Result = (rU/rS) 100
System suitability solution: Prepare a solution containing 1 mg/mL of USP Calcitonin Salmon RS. Combine equal volumes of this solution with USP Calcitonin Salmon Related Compound B RS. To 1 mL of this mixture, add 100 L of Buffer C. Sample solution: 0.5 mg/mL of Calcitonin Salmon. To 1 mL of this solution, add 100 mL of Buffer C. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 20-cm; packing L9 Flow rate: 1.2 mL/min Injection size: 50 L System suitability Sample: System suitability solution [NOTEThe relative retention times for [1,7-bis(3-sulpho-lalanine)]calcitonin salmon-glycine, [1,7-bis(3-sulpho-lalanine)]calcitonin salmon, and calcitonin salmon related compound B (calcitonin salmon-glycine) are 0.4, 0.6, and 0.9, respectively; and the retention time for calcitonin salmon is about 9 min.] Suitability requirements Resolution: NLT 3.0 between calcitonin salmon and calcitonin salmon related compound B Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Calcitonin Salmon taken: Result = (rU/rS) 100 rU rS = peak response for each impurity = sum of the responses of all peaks
USP 32
Acceptance criteria Individual impurities:
Name Calcitonin Salmon Related Compound B [1, 7 bis(3-sulphoL-alanine)] Calcitonin Salmon-glycine [1, 7 bis(3-sulphoL-alanine)] Calcitonin Salmon Any other impurity

0.91.1; lysine, 1.82.2; serine, 3.24.2; threonine, 4.25.2; tyrosine, 0.71.1; half cystine, 1.42.1. PEPTIDE MAPPING (See Biotechnology-Derived ArticlesPeptide Mapping 1055.) [NOTEThis test needs to be performed only on material produced using rDNA technology.] Solution A: Trifluoroacetic acid and water (1:1000) Solution B: Acetonitrile, trifluoroacetic acid, and water (800:0.85:200) Mobile phase: See gradient table below.
Time (min) 0.6 0.2 0 50 60 0.1 60.1 65.1 65.2 80.2 Solution A (%) 100 65 40 0 0 100 100 Solution B (%) 0 35 60 100 100 0 0
See Impurity Table.

Limit NMT (%) 0.6
Relative Retention Time 0.9
0.4
0.2
SPECIFIC TESTS AMINO ACID PROFILE (See Biotechnology-Derived ArticlesAmino Acid Analysis 1052.) [NOTEThis test is performed only on material of synthetic origin. The concentration of amino acids in the Internal standard solution, the Standard stock solution, and the Standard solution and the amount of material used to prepare the Sample solution can be adjusted depending on the method used for amino acid analysis. The concentrations given are based on analysis using Method 1.] Internal standard solution: 1 mM solution of aminobutyric acid Standard stock solution: 2.5 mM ammonia and the L form of lysine, histidine, arginine, aspartic acid, threonine, serine, proline, valine, glutamic acid, glycine, leucine, tyrosine, and L-cystine, in 0.1 M hydrochloric acid Standard solution: Transfer 5 mL of the Internal standard solution and 2 mL of the Standard stock solution into a 50mL volumetric flask, and dilute with 0.1 M hydrochloric acid to volume. Sample solution: Place 1.5 mg of Calcitonin Salmon into a heavy-wall ignition tube, add 1.0 mL of 6 N hydrochloric acid, allow to cool, immerse the lower half of the tube in a freezing mixture until the contents are frozen, evacuate to approximately 10 mm of Hg, purge with nitrogen (repeat the evacuation and nitrogen purge three times), and seal the tube while it is under a 10 mm of Hg vacuum. Heat for 16 h at 110 to 115 in an air oven. Cool, open the tube, dry in a vacuum desiccator, remove the contents, and allow to cool to room temperature. Dissolve in 0.1 M hydrochloric acid, transfer to a 10-mL volumetric flask, add 1 mL of Internal standard solution, and dilute with 0.1 M hydrochloric acid to volume. Analysis Samples: Standard solution and Sample solution Standardize the amino acid analyzer, using the Standard solution. Inject the Sample solution into the amino acid analyzer, and determine the relative proportion of amino acids. Express the content of each amino acid in moles, using an internal standard calibration technique. Calculate the relative proportions of the amino acids by taking as equivalent to 1 the sum of the number of moles of aspartic acid, glutamic acid, proline, glycine, valine, leucine, histidine, arginine, and lysine divided by 20. Acceptance criteria: The requirements are met if the values fall within the following limits: aspartic acid, 1.82.2; glutamic acid, 2.73.3; proline, 1.72.3; glycine, 2.73.3; valine, 0.91.1; leucine, 4.55.3; histidine, 0.91.1; arginine,
Trypsin solution: Freshly prepare a solution containing 0.1 mg/mL of trypsin (previously treated with L- 1-tosylamido-2phenylethyl chloromethyl ketone (TPCK) to remove chymotrypsin activity). Tris buffer: 1 M tris(hydroxymethyl) aminomethane, 10 mM calcium chloride, adjust with hydrochloric acid to a pH of 8.0 Stopping solution: Water and trifluoroacetic acid (1:1) Standard solution: 1.0 mg/mL of USP Calcitonin Salmon RS. Transfer 1 mL of this solution to a clean vial. Add 100 mL of Tris buffer and 50 mL of Trypsin solution, mix, and incubate at 2 to 8 for 1620 h. Quench the digestion by adding 10 mL of Stopping solution. Sample solution: 1.0 mg/mL of Calcitonin Salmon. Transfer 1 mL of this solution to a clean vial. Add 100 mL of Tris buffer and 50 mL of Trypsin solution, mix, and incubate at 2 to 8 for 1620 h. Quench the digestion by adding 10 mL of Stopping solution. Chromatographic system (See Chromatography, 621 System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L Analysis Samples: Standard solution and Sample solution [NOTECondition the chromatographic system by running a blank gradient program prior to injecting the digests.] Acceptance criteria: The chromatographic profile of the Sample solution is similar to that of the Standard solution BIOIDENTITY RPMI 1640 with L-glutamine: Prepare a mixture of the ingredients, in the quantities shown below, in sufficient water to obtain 1 L of RPMI 1640 with L-glutamine solution, and sterilize by filtration.
Calcium Nitrate Potassium Chloride Magnesium Sulfate, Anhydrous Potassium Chloride Sodium Chloride Sodium Phosphate, Dibasic, Anhydrous Sodium Bicarbonate 100.00 mg 400.00 mg 48.84 mg 400 mg 6000 mg 800 mg 2000 mg
12

10 mg 200 mg 50 mg 20 mg 65 mg 20 mg 300 mg 15 mg 20 mg 50 mg 50 mg 40 mg 15 mg 15 mg 20 mg 30 mg 20 mg 5 mg 29 mg 20 mg 0.2 mg 3 mg 0.25 mg 1 mg 35 mg 1 mg 1 mg 1 mg 0.2 mg 1 mg 0.005 mg Pantothenate Acid Dihydrochloride Acid
USP 32
Positive control solution: 1 ng/mL of USP Calcitonin Salmon RS, from Standard stock solution in Medium B Negative control solution: Medium B Standard solution A: 0.1 ng of USP Calcitonin Salmon RS, from Standard stock solution in Medium B Standard solution B: 33 pg/mL USP Calcitonin Salmon RS, from Standard solution A in Medium B Standard solution C: 11 pg/mL of USP Calcitonin Salmon RS, from Standard solution B in Medium B Standard solution D: 3.7 pg/mL USP Calcitonin Salmon RS, from Standard solution C in Medium B Sample stock solution: 20 g/mL of Calcitonin Salmon in Solution B Sample solution A: 0.1 ng/mL of Calcitonin Salmon, from Sample stock solution in Medium B Sample solution B: 33 pg/mL of Calcitonin Salmon, from Sample solution A in Medium B Sample solution C: 11 pg/mL of Calcitonin Salmon, from Sample solution B in Medium B Sample solution D: 3.7 pg/mL of Calcitonin Salmon, from Sample solution C in Medium B Cell culture preparation: Prepare a cell culture of the human mammary tumor cell line T-47D. Cells are propagated using Medium A at 37 and 5% carbon dioxide. The medium is changed every 2 days, and cells are passaged every 5 to 9 days, using TrypsinEDTA solution with a 1:4 subculture. Cell suspension: For the test, use a cell culture that is 59 days old. Remove the cell culture medium from the flask by aspiration, add 10 mL of Solution D, and rock the culture flask to rinse the entire monolayer. Remove the liquid by aspiration, add 2 mL of Solution C, spread over the entire monolayer, allow to stand for 35 min, and add 10 mL of Medium A. Homogenize the cell suspension using a pipet, transfer to a 15-mL polypropylene tube, centrifuge at about 220 g for 5 min, pour off the supernatant, and resuspend the cell pellet in 10 mL of Medium A. Count the cells, and adjust the cell density through dilution, using Medium A, to 2.5 104 cells/mL. Analysis: Place 200 L of the Cell suspension into each well of a 96-well culture plate (the tissue culture plate), and incubate for 1824 h at 37 and 5% carbon dioxide. Fill each well of an empty round-bottomed 96-well culture plate (the prepared plate) with 150 L of one of the following solutions: Positive control solution, Negative control solution, Standard solutions AD, and Sample solutions AD, so that each solution fills at least five wells on the prepared plate. After incubation, remove the culture medium from the tissue culture plate. Using an 8-channel or 12-channel pipet, rapidly transfer 100 L of solution from each well of the prepared plate to each well of the tissue culture plate. Incubate for 15 min at ambient temperature, remove the solution from each well, stop stimulation by immediately adding an appropriate cell-lysis buffer, and quantitate cAMP produced within the cells, using a validated kit. Perform the test three times, using three different 96-well culture plates. [NOTESome kits include a cell-lysis reagent and a sequestering agent for the cell-lysis reagent. The range of the test kit is between 0.05 ng and 10 ng/mL of cAMP. The number of cells used in the assay may vary depending on the validated kit used to quantitate cAMP.] Potency is determined by a 3-dose, 6-point parallel-line assay, using standard statistical methods. The calculation is carried out using both the lower three concentrations and the upper three concentrations. For the assay to be valid, the requirements for regression and parallelism must be met. If the requirements for validity are met to the same extent in both assessments (the lower and the higher assessments), the final result is determined from the concentration range that shows the higher value when the common slope is divided by the mean square error.
Glycine
L-Arginine L-Asparagine L-Aspartic L-Cystine
L-Glutamic L-Histidine
L-Glutamine L-Hydroxyproline L-Isoleucine L-Leucine L-Lysine
Hydrochloride
L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine
Disodium Salt Dihydrate
Biotin Choline Chloride

D-Calcium
Folic Acid i-Inositol Niacinamide para-Aminobenzoic Acid Pyridoxine Hydrochloride Riboflavin Thiamine Hydrochloride Vitamin B12
Medium A (growth medium): Using aseptic technique, prepare the following tissue culture medium.
RPMI 1640 with L-glutamine Fetal bovine serum 1 M HEPES Penicillin/streptomycin solution (10,000 IU per mL/10 mg per mL) Human insulin Hydrocortisone 500 mL 50 mL 5 mL 5 mL 10 IU 0.5 mg
Medium B (stimulation medium): Dissolve 5 g of albumin bovine serum (BSA) in 500 mL of 2 mM RPMI 1640 with Lglutamine. Solution A: Dissolve 500 mg of albumin bovine serum in 25 mL of water. [NOTEUse within 1 day.] Solution B: Add 25 mL of 0.1 M formic acid and 5 mL of Solution A to a 50-mL volumetric flask, dilute with water to volume, and mix. [NOTEUse within 2 days.] Solution C: Prepare a sterile filtered solution containing 0.25% trypsin and 0.53 mM EDTA. Solution D: Dissolve 8 g of sodium chloride, 1.15 g of dibasic sodium phosphate, 0.2 g of monobasic potassium phosphate, 0.2 g of potassium chloride, 0.1 g of calcium chloride, and 0.1 g of magnesium chloride in 1 L of water. Standard stock solution: 20 g of USP Calcitonin Salmon RS in Solution B.
USP 32
Acceptance criteria: The potency levels determined from all three performances of the test are homogeneous, and the confidence limits for all three determinations are between 64% and 156% of the calculated potency. Change to read: MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62 Sample: 0.2 g Acceptance criteria: The total aerobic microbial count NMT 100 cfu/g vand the total combined molds and yeasts is NMT 100 cfu/gvUSP32 WATER DETERMINATION, Method Ic 921: NMT 10% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store in a refrigerator or maintain in a frozen state, protected from light. LABELING: The labeling states that the material is synthetic or of recombinant DNA origin. USP REFERENCE STANDARD 11 USP Calcitonin Salmon RS USP Calcitonin Salmon Related Compound A RS (N-acetylcys1 -calcitonin salmon) USP Calcitonin Salmon Related Compound B RS (calcitonin salmon-glycine)

0.4 mL of Solution A, and add 0.1 mL of the Standard solution. System suitability solution: System suitability stock solution and Solution A (1:9) Sample solution: Use the Injection. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 65 Flow rate: 1 mL/min Injection size: 200 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for calcitonin salmon and calcitonin salmon related compound A are 1.0 and 1.15, respectively.] Suitability requirements Resolution: NLT 3 between calcitonin salmon and calcitonin salmon related compound A Tailing factor: NMT 2.5 Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the potency, in USP Calcitonin Salmon Units/mL, in the portion of Injection taken: Result = (rU/rS) (CS/CU) 100 = peak area response of the Sample solution = peak area response of the Standard solution = concentration of USP Calcitonin Salmon RS in the Standard solution (mg/mL) = nominal concentration of calcitonin salmon in CU the Sample solution (mg/mL) Acceptance criteria: 80.0%110.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.625 USP Endotoxin Unit/USP Calcitonin Salmon Unit STERILITY TESTS 71: Meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 3.94.5 INJECTIONS 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. Avoid freezing. Store in a refrigerator. LABELING: Label it to indicate the activity in USP Calcitonin Salmon Units per mL. The labeling states that the material is synthetic. Label it to state that it is to be stored in a refrigerator, and that freezing is to be avoided. USP REFERENCE STANDARDS 11 USP Calcitonin Salmon RS USP Calcitonin Salmon Related Compound A RS USP Endotoxin RS rU rS CS
Calcitonin Salmon Injection

(Comment on this Monograph)id=m11350=Calcitonin Salmon Injection=Ca-Chl-Monos.pdf) DEFINITION Calcitonin Salmon Injection is a sterile solution of Calcitonin Salmon in a suitable diluent. Each mL of Calcitonin Salmon Injection possesses an activity of NLT 80% and NMT 110% of that stated on the label. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 3.26 mg/mL of tetramethylammonium hydroxide pentahydrate in water and acetonitrile (9:1). Adjust with phosphoric acid to a pH of 2.5. Solution B: 1.45 mg/mL of tetramethylammonium hydroxide pentahydrate in water and acetonitrile (2:3). Adjust with phosphoric acid to a pH of 2.5. Mobile phase: See the gradient table below.
Calcitonin Salmon Nasal Solution

(Comment on this Monograph)id=m11342=Calcitonin Salmon Nasal Solution=Ca-Chl-Monos.pdf) DEFINITION Calcitonin Salmon Nasal Solution is a solution of Calcitonin Salmon in a suitable diluent. It contains suitable preservatives,
Standard stock solution: 1.0 mg/mL of USP Calcitonin Salmon RS in Solution A Standard solution: 0.1 mg/mL of USP Calcitonin Salmon RS from Standard stock solution in Solution A System suitability stock solution: Dissolve the contents of a vial of USP Calcitonin Salmon Related Compound A RS in
14

CU
USP 32
= nominal concentration of calcitonin salmon in the Sample solution (mg/mL) Acceptance criteria: 80.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: Total aerobic microbial count: NMT 100 cfu/g Total combined molds and yeast count: NMT 50 cfu/g. It meets the requirements for absence of Staphylococcus aureus and Pseudomonas aeruginosa. PH 791: 3.54.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in containers suitable for spraying the contents into the nasal cavities in a controlled individualized dosage. Store unopened containers in a refrigerator, and opened containers at room temperature. LABELING: Label it to indicate that it is for intranasal administration only. The labeling also states that it has been prepared either with Calcitonin Salmon of synthetic origin or Calcitonin Salmon of rDNA origin. Label it to state that it is to be stored in a refrigerator, and that freezing is to be avoided. Label it to indicate the activity in USP Calcitonin Salmon Units/mL. USP REFERENCE STANDARDS 11 USP Calcitonin Salmon RS USP Calcitonin Salmon Related Compound A RS
and is packaged in a form suitable for nasal administration so that the required dosage can be controlled as required. Each mL of Calcitonin Salmon Nasal Solution possesses an activity of NLT 80% and NMT 110% of that stated on the label. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 3.26 mg/mL of tetramethylammonium hydroxide pentahydrate in water and acetonitrile (9:1). Adjust with phosphoric acid to a pH of 2.5. Solution B: 1.45 mg/mL of tetramethylammonium hydroxide pentahydrate in acetonitrile and water (3:2). Adjust with phosphoric acid to a pH of 2.5. Mobile phase: See the gradient table below.
Standard stock solution: 1.0 mg/mL of USP Calcitonin Salmon RS in Solution A Standard solution: 0.1 mg/mL of USP Calcitonin Salmon RS from Standard stock solution in Solution A System suitability stock solution: Dissolve the contents of a vial of USP Calcitonin Salmon Related Compound A RS in 0.4 mL of Solution A, and add 0.1 mL of the Standard solution. System suitability solution: System suitability stock solution and Solution A (1:9) Diluent: 7.5 mg/mL of sodium chloride, 2 mg/mL of sodium acetate, and 2 mg/mL of glacial acetic acid in water Sample solution: Nasal Solution in Diluent (1:9) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 65 Flow rate: 1 mL/min Injection size: 200 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for calcitonin salmon and calcitonin salmon related compound A are 1.0 and 1.15, respectively.] Suitability requirements Resolution: NLT 3 between calcitonin salmon and calcitonin salmon related compound A Tailing factor: NMT 2.5 Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the potency, in USP Calcitonin Salmon Units/mL, in the portion of Nasal Solution taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak area response of the Sample solution = peak area response of the Standard solution = concentration of USP Calcitonin Salmon RS in the Standard solution (mg/mL)
Calcium Acetate
(Comment on this Monograph)id=m11400=Calcium Acetate=Ca-Chl-Monos.pdf)
C4H6CaO4 Acetic acid, calcium salt; Calcium acetate [62-54-4].
158.17
DEFINITION Calcium Acetate contains NLT 99.0% and NMT 100.5% of C4H6CaO4, calculated on the anhydrous basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Calcium, 191 AND Acetate, 191 Sample solution: 50 mg/mL Acceptance criteria: Meets the requirements ASSAY PROCEDURE Sample solution: 50 mg/mL Acceptance criteria: Meets the requirements Sample: 300 mg Analysis: Dissolve the Sample in 150 mL of water containing 2 mL of 3 N hydrochloric acid. While stirring, add about 30 mL of 0.05 M edetate disodium VS from a 50mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 7.909 mg of C4H6CaO4. Acceptance criteria: 99.0%100.5%, calculated on the anhydrous basis
USP 32
IMPURITIES Inorganic Impurities LIMIT OF FLUORIDE [NOTEPrepare and store all solutions in plastic containers.] Solution A: 294 mg/mL of sodium citrate dihydrate in water Standard stock solution: 1.1052 mg/mL of USP Sodium Fluoride RS Standard solution: Combine 20.0 mL of Standard stock solution with 50.0 mL of Solution A and dilute to 100.0 mL with water. Electrode system: Use a fluoride-specific, ion-indicating electrode and a silversilver chloride reference electrode connected to a pH meter capable of measuring potentials with a minimum reproducibility of 0.2 mV (see pH 791). Standard response line: Transfer 50.0 mL of Solution A and 2.0 mL of hydrochloric acid to a beaker, and add water to make 100 mL. Add a plastic-coated stirring bar, insert the electrodes into the solution, stir for 15 min, and read the potential, in mV. Continue stirring, and at 5-min intervals add 100, 100, 300, and 500 L of Standard solution, reading the potential 5 min after each addition. Plot the logarithms of the cumulative fluoride ion concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus potential, in mV. Sample solution: Transfer 2.0 g of the Calcium Acetate to a beaker containing a plastic-coated stirring bar, add 20.0 mL of water and 2.0 mL of hydrochloric acid, and stir until dissolved. Add 50.0 mL of Solution A and sufficient water to make 100 mL. Analysis: Rinse and dry the electrodes, insert them into the Sample solution, stir for 5 min, and read the potential, in mV. From the measured potential and the Standard response line determine the concentration, C, in g/mL, of fluoride ion in the Sample solution. Calculate the amount of fluoride (ppm) in the specimen taken by multiplying C by 0.005. Acceptance criteria: 50 ppm CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion shows no more chloride than corresponds to 0.70 mL of 0.020 N hydrochloric acid (0.05%). CHLORIDE AND SULFATE, Sulfate 221: A 0.25-g portion shows no more sulfate than corresponds to 0.15 mL of 0.020 N sulfuric acid (0.06%). ARSENIC, Method I 211: NMT 3 ppm LEAD 251: NMT 10 ppm HEAVY METALS, Method I 231 Sample solution: Dissolve 0.8 g of Calcium Acetate in 20.0 mL of water, add 3.0 mL of glacial acetic acid, dilute with water to 25 mL, and adjust with glacial acetic acid to a pH of 3.84.0, measured with a pH meter. Monitor preparation: Prepare as directed for Sample solution, 2.0 mL of Standard lead solution being added. Acceptance criteria: NMT 25 ppm LIMIT OF NITRATE Sample solution: 100 mg/mL of Calcium Acetate in water Analysis: To 10 mL of the Sample solution, add 5 mg of sodium chloride, 0.05 mL of indigo carmine TS, and, with stirring, 10 mL of nitrogen-free sulfuric acid. Acceptance criteria: The blue color persists for NLT 10 min. LIMIT OF ALUMINUM [NOTEUse where it is labeled as intended for parenteral use or for use in hemodialysis or peritoneal dialysis.] Solution A: Dissolve 200 mg/mL of ammonium acetate in water, and adjust with glacial acetic acid to a pH of 6.0 prior to final dilution. Aluminum standard stock solution: Treat some aluminum wire with 6 N hydrochloric acid at 80 for a few min.
Official Monographs / Calcium 15

Dissolve about 100 mg of the treated wire in a mixture of 10 mL of hydrochloric acid and 2 mL of nitric acid by heating at about 80 for approximately 30 min. Continue heating until the volume is reduced to about 4 mL. Cool to room temperature, and add 4 mL of water. Evaporate to about 2 mL by heating. Cool, and transfer this solution, with the aid of water, to a 100-mL volumetric flask, dilute with water to volume, and mix. [NOTEEquivalent to 1 mg/mL of Aluminum.] Aluminum standard solution: 1.0 g/mL of Aluminum, from Aluminum standard stock solution in water. Standard solution: Prepare a solution containing 2.0 mL of Aluminum standard solution, 5 mL of Solution A, and 48 mL of water, and extract this solution with successive portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50mL volumetric flask. Dilute the combined extracts with chloroform to volume. Sample solution: Dissolve 1.0 g of Calcium Acetate in 50 mL of water and add 5 mL of Solution A. Extract this solution with successive portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50-mL volumetric flask. Dilute the combined extracts with chloroform to volume. Blank: Prepare a solution containing 50 mL of water and 5 mL of Solution A, and extract this solution with successive portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50mL volumetric flask. Dilute the combined extracts with chloroform to volume. Spectrometric conditions Mode: fluorescence Excitation wavelength: 392 nm Emission wavelength: 518 nm Analysis Samples: Sample solution and Standard solution. Acceptance criteria: The fluorescence of the Sample solution is NMT that of the Standard solution (2 ppm). LIMIT OF BARIUM: [NOTEUse where it is labeled as intended for use in hemodialysis or peritoneal dialysis.] Barium chloride solution: 0.758 mg/mL of anhydrous barium chloride [NOTEThis solution contains 500 g/mL of barium.] Standard solution: To a fifth tube, add 1 g of ammonium acetate, 2 mL of 1 N hydrochloric acid, 3.0 mL of Barium chloride solution, and sufficient water to bring the volume to 40 mL. Sample stock solution: 250 mg/mL of Calcium Acetate and 25 mg/mL of ammonium acetate in 1 N hydrochloric acid. The pH of this solution is 4.55.5. [NOTEFilter and cover the solution.] Sample solution: To four separate tubes, add 1.0, 1.5, 2.0, and 2.5 mL of Barium chloride solution. To each tube, add a sufficient volume of the Sample stock solution to bring the volume to 40 mL. Solution A: Ammonium sulfate solution (1 in 10) Analysis: To the Sample solutions and the Standard solution add, with brisk stirring, 3.0 mL of Solution A, and allow to stand for 20 min. Acceptance criteria: The Sample solutions containing 1.0 and 1.5 mL of Barium chloride solution remain clear or are only faintly turbid. The Sample solution containing 2.0 mL of Barium chloride solution is not more turbid than the Standard solution. LIMIT OF MAGNESIUM: [NOTEUse where it is labeled as intended for use in hemodialysis or peritoneal dialysis.] [NOTEThe Standard solution and the Sample solutions may be modified, if necessary, to obtain solutions of suitable concentrations, adaptable to the linear or working range of the instrument.]
16
Calcium / Official Monographs
USP 32
Acceptance criteria: NMT 0.05% LIMIT OF SODIUM: [NOTEUse where it is labeled as intended for use in hemodialysis or peritoneal dialysis.] [NOTEThe Standard solution and the Sample solutions may be modified, if necessary, to obtain solutions of suitable concentrations, adaptable to the linear or working range of the instrument.] Standard stock solution A: 25.42 mg/mL of sodium chloride, using sodium chloride previously dried at 105 for 2 h [NOTEThis solution contains 10.0 mg/mL of sodium.] Standard stock solution B: 250 g/mL of sodium: from Standard stock solution Standard solution A: Dilute 20.0 mL of Sample solution to 25.0 mL with water Standard solution B: Dilute 2.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Standard solution C: Dilute 4.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Sample solution: 10 mg/mL Blank: water Spectometric conditions (see Spectrophotometry and Light-Scattering 851) Mode: Atomic absorption spectroscopy Source: air-acetylene flame Detector sodium hollow-cathode lamp Analytical wavelength: 589.0 nm Analysis: Plot the absorbances of the treated Sample solutions versus their contents of sodium, in g/mL, draw the straight line best fitting the three points, and extrapolate the line until it intercepts the concentration axis. From the intercept determine the amount, in g, of sodium in each mL of the Sample solution. Calculate the percentage of sodium in the specimen by multiplying this value by 0.0125. Acceptance criteria: NMT 0.5% LIMIT OF STRONTIUM: [NOTEUse where it is labeled as intended for use in hemodialysis or peritoneal dialysis.] [NOTEThe Standard solution and the Sample solutions may be modified, if necessary, to obtain solutions of suitable concentrations, adaptable to the linear or working range of the instrument.] Standard stock solution A: 2.45 mg/mL of strontium acetate in water. [NOTEThis solution contains 1000 g/mL of strontium.] Standard stock solution B: 50.0 g/mL of strontium, from Standard stock solution in water Standard solution A: Dilute 20.0 mL of Sample solution to 25.0 mL with water Standard solution B: Dilute 2.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Standard solution C: Dilute 4.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Sample solution: 20 mg/mL Blank: water Spectometric conditions (see Spectrophotometry and Light-Scattering 851) Mode: Atomic absorption spectroscopy Source: Nitrous oxide-acetylene Detector strontium hollow-cathode lamp Analytical wavelength: 460.7 nm Analysis: Plot the absorbances of the treated Sample solutions versus their contents of strontium, in g/mL, draw the straight line best fitting the three points, and extrapolate the line until it intercepts the concentration axis. From the intercept determine the amount, in g, of strontium in each mL of the Sample solution.
Standard stock solution A: 1.516 mg/mL of magnesium oxide in 1 N nitric acid [NOTEThis solution contains 1000 g/mL of magnesium.] Standard stock solution B: 5.0 g/mL of magnesium, from Standard stock solution Standard solution A: Dilute 20.0 mL of Sample solution to 25.0 mL with water Standard solution B: Dilute 2.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Standard solution C: Dilute 4.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Sample solution: 2 mg/mL of Calcium Acetate Blank: water Spectometric conditions Mode: Atomic absorption spectroscopy Source: Air-acetylene flame Detector magnesium hollow-cathode lamp Analytical wavelength: 285.2 Analysis Samples: Standard solution A, Standard solution B, and Standard solution C Plot the absorbances of the Standard solutions versus their contents of magnesium, in g/mL, draw the straight line best fitting the three points, and extrapolate the line until it intercepts the concentration axis. From the intercept determine the amount, in g, of magnesium in each mL of the Sample solution. Calculate the percentage of magnesium in the specimen by multiplying this value by 0.0625. Acceptance criteria: NMT 0.05% LIMIT OF POTASSIUM: [NOTEUse where it is labeled as intended for use in hemodialysis or peritoneal dialysis.] [NOTEThe Standard solution and the Sample solutions may be modified, if necessary, to obtain solutions of suitable concentrations, adaptable to the linear or working range of the instrument.] Standard stock solution A: 23.836 mg/mL of potassium chloride, using potassium chloride previously dried at 105 for 2 h [NOTEThis solution contains 12.5 mg/mL of potassium.] Standard stock solution B: 31.25 g/mL of potassium, from Standard stock solution Standard solution A: Dilute 20.0 mL of Sample solution to 25.0 mL with water Standard solution B: Dilute 2.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Standard solution C: Dilute 4.0 mL of Standard stock solution B and 20.0 mL of Sample solution to 25.0 mL with water Sample solution: 12.5 mg/mL Blank: water Spectometric conditions (see Spectrophotometry and Light-Scattering 851) Mode: Atomic absorption spectroscopy Source: Air-acetylene flame Detector potassium hollow-cathode lamp Analytical wavelength: 766.7 nm Analysis Samples: Standard solution A, Standard solution B, and Standard Solution C. Plot the absorbances of the Standard solutions versus their contents of potassium, in g/mL, draw the straight line best fitting the three points, and extrapolate the line until it intercepts the concentration axis. From the intercept determine the amount, in g, of potassium in each mL of the Sample solution. Calculate the percentage of potassium in the specimen by multiplying this value by 0.01.
USP 32
Calculate the percentage of strontium in the specimen by multiplying this value by 0.00625. Acceptance criteria: NMT 0.05% Organic Impurities PROCEDURE: READILY OXIDIZABLE SUBSTANCES Sample solution: 20 mg/mL of Calcium acetate in boiling water Analysis: Add a few glass beads to 100 mL of the Sample solution, 6 mL of 10 N sulfuric acid, and 0.3 mL of 1 N potassium permanganate, mix, boil gently for 5 min, and allow the precipitate to settle. Acceptance criteria: The pink color in the supernatant is not completely discharged. SPECIFIC TESTS PH 791 Sample solution: 50 mg/mL Acceptance criteria: 6.39.6 WATER DETERMINATION, Method I 921 Sample: 0.7 g Analysis: Proceed as directed, adding 20.0 mL of glacial acetic acid to the titration vessel in addition to the methanol. Acceptance criteria: NMT 7.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where Calcium Acetate is intended for use in hemodialysis or peritoneal dialysis, it is so labeled.

Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to Standard solution. Acceptance criteria: NLT 80% (Q) of the labeled amount of C4H6CaO4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Inorganic Impurities LIMIT OF ALUMINUM [NOTEUse where it is labeled as intended for parenteral use or for use in hemodialysis or peritoneal dialysis.] Solution A: Dissolve 200 mg/mL of ammonium acetate in water, and adjust with glacial acetic acid to a pH of 6.0 prior to final dilution. Aluminum standard stock solution: Treat some aluminum wire with 6 N hydrochloric acid at 80 for a few min. Dissolve about 100 mg of the treated wire in a mixture of 10 mL of hydrochloric acid and 2 mL of nitric acid by heating at about 80 for approximately 30 min. Continue heating until the volume is reduced to about 4 mL. Cool to room temperature, and add 4 mL of water. Evaporate to about 2 mL by heating. Cool, and transfer this solution, with the aid of water, to a 100-mL volumetric flask, and dilute with water to volume. [NOTEEquivalent to 1 mg/mL of aluminum] Aluminum standard solution 1.0 g/mL of aluminum from Aluminum standard stock solution in water Standard solution: Prepare a solution containing 2.0 mL of Aluminum standard stock solution, 5 mL of Solution A, and 48 mL of water, and extract this solution with successive portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50mL volumetric flask. Dilute the combined extracts with chloroform to volume. Sample solution: Prepare a solution containing an amount of powdered Tablets equivalent to 1.0 g of calcium acetate (powder NLT 20 Tablets), 50 mL of water, and 5 mL of Solution A. Extract this solution with successive portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50-mL volumetric flask. Dilute the combined extracts with chloroform to volume. Blank: Prepare a solution containing 50 mL of water and 5 mL of Solution A, and extract as described for the Aluminum standard solution. Extract this solution with successive portions of 10, 10, and 5 mL of a 0.5% 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50mL volumetric flask. Dilute the combined extracts with chloroform to volume. Spectrometric conditions Mode: fluorescence Excitation wavelength: 392 nm Emission wavelength: 618 nm Analysis Samples: Sample solution and Standard solution Acceptance criteria: The fluorescence of the Sample solution does not exceed that of the Standard solution NMT 2 g/g of aluminum calcium acetate
Calcium Acetate Tablets

(Comment on this Monograph)id=m11405=Calcium Acetate Tablets=Ca-Chl-Monos.pdf) DEFINITION Calcium Acetate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of calcium acetate (C4H6CaO4). IDENTIFICATION IDENTIFICATION TESTSGENERAL, Calcium and Acetate 191 Sample solution: 100 mg/mL of calcium acetate, from powdered Tablets, in water Acceptance criteria: Meet the requirements of the tests ASSAY PROCEDURE Sample: Amount equivalent to 300 mg of calcium acetate from NLT 20 powdered Tablets Analysis: Dissolve the Sample in 150 mL of water containing 2 mL of 3 N hydrochloric acid. While stirring, add 30 mL of 0.05 M edetate disodium VS from a 50-mL buret, and add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue. Continue the titration with the 0.05 M edetate disodium VS to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 7.909 mg of C4H6CaO4. Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Spectrometric conditions Mode: Atomic absorption spectrometry Detector: calcium hollow-cathode tube Analytical wavelength: 422.8 nm Standard solution: Calcium at a known concentration in Medium
18
USP 32
Calculate the concentration, in ppm, of fluoride in the Calcium Ascorbate taken: Result = (C/CU) x F1
Calcium Ascorbate
(Comment on this Monograph)id=m11410=Calcium Ascorbate=Ca-Chl-Monos.pdf) C12H14CaO12 2H2O 426.34 DEFINITION Calcium Ascorbate contains NLT 98.0% and NMT 101.0% of C12H14CaO12 2H2O, calculated on the as-is basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. A 100 mg/mL solution of Calcium Ascorbate decolorizes a 100 mg/mL solution of dichlorophenol-indophenol C. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample: 100 mg/mL ASSAY PROCEDURE Sample solution: 60 mg/mL of Calcium Ascorbate in 50 mL of water Analysis: Immediately titrate 50 mL of Sample solution with 0.1 N iodine VS, adding 3 mL of starch TS as the endpoint is approached. Each mL of 0.1 N iodine is equivalent to 10.66 mg of C12H14CaO12 2H2O. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities ARSENIC, Method I 211: 3 ppm HEAVY METALS, Method II 231: NMT 10 ppm LIMIT OF FLUORIDE [NOTEPrepare and store all solutions in plastic containers.] Solution A: 294 mg/mL of sodium citrate dihydrate Standard stock solution: 1.1052 mg/mL of USP Sodium Fluoride RS Standard solution: Combine 20.0 mL of Standard stock solution with 50.0 mL of Solution A, and dilute with water to 100 mL Sample solution: Transfer 2.0 g of Calcium Ascorbate to a beaker containing a plastic-coated stirring bar, add 20 mL of water and 2.0 mL of hydrochloric acid, and stir until dissolved. Add 50.0 mL of Solution A and sufficient water to make 100 mL. Electrode system: Use a fluoride-specific, ion-indicating electrode and a silversilver chloride reference electrode connected to a pH meter capable of measuring potentials with a minimum reproducibility of 0.2 mV (see pH 791). Analysis Samples: Standard solution and Sample solution Standard response line: Transfer 50.0 mL of Solution A and 2.0 mL of hydrochloric acid to a beaker, and add water to make 100 mL. Add a plastic-coated stirring bar, insert the electrodes into the solution, stir for 15 min, and read the potential, in mV. Continue stirring, and at 5-min intervals add 100, 100, 300, and 500 L of Standard solution, reading the potential 5 min after each addition. Plot the logarithms of the cumulative fluoride ion concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus potential, in mV. Rinse and dry the electrodes, insert them into the Sample solution, stir for 5 min, and read the potential, in mV. From the measured potential and the Standard response line determine the concentration, C (in g/mL) of fluoride ion in the Sample solution.
= measured concentration of fluoride ion in the Sample solution (g/mL) = concentration of Calcium Ascorbate in the CU Sample solution (mg/mL) F1 = unit conversion factor, 1 ppm-mg/g Acceptance criteria: NMT 10 ppm SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +95 to +97, from optical rotation measurements made immediately following the preparation of the Sample solution Sample solution: 50 mg/mL, in carbon dioxide-free water PH 791: 6.87.4, in a 100 mg/mL solution LOSS ON DRYING 731: Dry 3 g of it at 105 for 2 h: it loses NMT 0.1% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Calcium Ascorbate RS USP Sodium Fluoride RS
Calcium Carbonate
(Comment on this Monograph)id=m11430=Calcium Carbonate=Ca-Chl-Monos.pdf) CaCO3 Carbonic acid, calcium salt (1:1); Calcium carbonate (1:1) [471-34-1]. 100.09
DEFINITION Calcium Carbonate, dried at 200 for 4 h, contains calcium equivalent to NLT 98.0% and NMT 100.5% IDENTIFICATION IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of acetic acid to it produces effervescence (presence of carbonate), and the resulting solution, after boiling, meets the requirements of the tests. ASSAY PROCEDURE Sample: 200 mg of Calcium Carbonate, previously dried at 200 for 4 h Analysis: Transfer the Sample to a 250-mL beaker. Moisten thoroughly with a few mL of water, and add, dropwise, sufficient 3 N hydrochloric acid to dissolve. Add 100 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Titrate with 0.05 M edetate disodium VS until the solution is a distinct blue in color. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities LIMIT OF FLUORIDE [NOTEPrepare and store all solutions in plastic containers.] Solution A: 294 mg/mL of sodium citrate dihydrate in water Sample: 2.0 g Standard stock solution: 1.1052 mg/mL of USP Sodium Fluoride RS in water
USP 32
Standard solution: Combine 20.0 mL of Standard stock solution with 50.0 mL of Solution A and dilute to 100.0 mL with water Electrode system: Use a fluoride-specific, ion-indicating electrode and a silversilver chloride reference electrode connected to a pH meter capable of measuring potentials with a minimum reproducibility of 0.2 mV (see pH 791). Standard response line: Transfer 50.0 mL of Buffer solution and 2.0 mL of hydrochloric acid to a beaker, and add water to make 100 mL. Add a plastic-coated stirring bar, insert the electrodes into the solution, stir for 15 min, and read the potential, in mV. Continue stirring, and at 5-min intervals add 100, 100, 300, and 500 L of Standard solution, reading the potential 5 min after each addition. Plot the logarithms of the cumulative fluoride ion concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus potential, in mV. Analysis: Transfer the Sample to a beaker containing a plastic-coated stirring bar, add 20 mL of water and 4.0 mL of hydrochloric acid, and stir until dissolved. Add 50.0 mL of Buffer solution and sufficient water to make 100 mL of test solution. Rinse and dry the electrodes, insert them into the Sample solution, stir for 5 min, and read the potential, in mV. From the measured potential and the Standard response line, determine the concentration, C, in g/mL, of fluoride ion in the Sample solution. Calculate the percentage of fluoride in the specimen taken by multiplying C by 0.005. Acceptance criteria: 50 ppm ACID-INSOLUBLE SUBSTANCES: Mix 5.0 g with 10 mL of water, and add hydrochloric acid, dropwise, with agitation, until it ceases to cause effervescence, then add water to make the mixture measure 200 mL, and filter. Wash the insoluble residue with water until the last washing shows no chloride, and ignite: the weight of the residue does not exceed 10 mg (0.2%). BARIUM: A platinum wire, dipped in the filtrate obtained in the test for Acid-Insoluble Substances and held in a nonluminous flame, does not impart a green color. ARSENIC, Method I 211 Sample solution: Slowly dissolve 1.0 g in 15 mL of hydrochloric acid, and dilute with water to 55 mL. Acceptance criteria: NMT 3 ppm. The resulting solution meets the requirements of the test, the addition of 20 mL of 7 N sulfuric acid specified under Procedure being omitted. LEAD 251 Sample solution: 1.0 g in 5 mL of water Analysis: To the Sample solution slowly add 8 mL of 3 N hydrochloric acid, evaporate on a steam bath to dryness, and dissolve the residue in 5 mL of water. Acceptance criteria: NMT 3 ppm IRON 241 Sample: 40 mg Spectrometric conditions Analytical wavelength: 530 nm Blank: Water Analysis: Dissolve the Sample in 5 mL of 2 N hydrochloric acid. Transfer to a beaker with the aid of water, and dilute with water to 10 mL. Prepare a Standard solution by transferring 4.0 mL of Standard iron solution, prepared as directed under Iron 241, to a beaker and by diluting with water to 10 mL. To each beaker, add 2 mL of citric acid solution (1 in 5) and 2 drops of thioglycolic acid, adjust to a pH of 9.5 0.1 with ammonia TS, dilute with water to 20 mL, mix, and allow to stand for 5 min. Dilute with water to 50 mL. Concomitantly determine the absorbances of the solutions from the Sample and the Standard solution.

Acceptance criteria: The absorbance of the solution from the specimen under test does not exceed that of the Standard solution (0.1%). MERCURY, Method IIa 261 Mercury stock solution and Standard mercury solution: Proceed as directed under Mercury 261. Standard solution: Pipet 2.0 mL of Standard mercury solution into a 100-mL beaker, and add 35 mL of water, 3 mL of hydrochloric acid, and 1 mL of Potassium permanganate solution. Cover the beaker with a watch glass, boil for a few s, and cool. Sample stock solution: 4.0 g in a 100-mL beaker, and cautiously dissolve in 14 mL of 6 N hydrochloric acid Sample solution: Transfer the Sample stock solution to a 100-mL beaker, and add 35 mL of water. Stir, and warm to assist solution, if necessary. Add 2 drops of phenolphthalein TS, and, as necessary, slowly neutralize with constant stirring, using 1 N sodium hydroxide or 1 N sulfuric acid. Add 3 mL of hydrochloric acid and 1 mL of Potassium permanganate solution. Cover the beaker with a watch glass, boil for a few s, and cool. Analysis Samples: Standard solution and Sample solution Proceed as directed in Mercury 261. Acceptance criteria: 0.5 ppm LIMIT OF MAGNESIUM and ALKALI SALTS Sample solution: 1.0 g Analysis: Mix the Sample with 35 mL of water. Carefully add 3 mL of hydrochloric acid, heat the solution, and boil for 1 min. Rapidly add 40 mL of oxalic acid TS, and stir vigorously until precipitation is well-established. Add immediately to the warm mixture 2 drops of methyl red TS and then 6 N ammonium hydroxide, dropwise, until the mixture is just alkaline. Cool to room temperature, transfer to a 100-mL graduated cylinder, dilute with water to 100 mL, mix, and allow to stand for 4 h or overnight. Filter, and to 50 mL of the clear filtrate in a platinum dish add 0.5 mL of sulfuric acid, and evaporate the mixture on a steam bath to a small volume. Carefully heat over a free flame to dryness, and continue heating to complete decomposition and volatilization of ammonium salts. Finally, ignite the residue to constant weight. Acceptance criteria: The weight of the residue is NMT 5 mg (NMT 1.0%). HEAVY METALS 231 Sample solution: Mix 1.0 g with 5 mL of water, slowly add 8 mL of 3 N hydrochloric acid, and evaporate on a steam bath to dryness. Dissolve the residue in 20 mL of water, filter, and add water to the filtrate to make 25 mL. Acceptance criteria: NMT 20 ppm SPECIFIC TESTS LOSS ON DRYING 731: 2.0% of its weight. Dry it at 200 for 4 h: it loses NMT
Calcium Carbonate Lozenges

(Comment on this Monograph)id=m11439=Calcium Carbonate Lozenges=Ca-Chl-Monos.pdf) DEFINITION Calcium Carbonate Lozenges contain NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3).
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Standard stock solution: 2.542 mg/mL of sodium chloride, previously dried at 105 for 2 h (equivalent to 1.00 mg/mL sodium) Standard solution A: 1.0 g/mL of sodium from Standard stock solution Standard solution B: 3.0 g/mL of sodium from Standard stock solution Standard solution C: 5.0 g/mL of sodium from Standard stock solution Sample solution: Sample stock solution in the Assay. Pass, if necessary, through a filter having a 0.5-m or finer porosity to obtain a clear solution. Transfer 10.0 mL of the clear solution to a 25-mL volumetric flask, and dilute with water to volume. Blank: Water Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Flame: Airacetylene Lamp: Sodium hollow-cathode Analytical wavelength: 589.6 nm Analysis Samples: Standard solution A, Standard solution B, Standard solution C, Sample solution, and Blank Plot the absorbances of the Standard solutions versus their contents of sodium, in g/mL, by drawing a straight line best fitting the three plotted points. From the graph determine the quantity, C, in g, of sodium in each mL of the Sample solution. Calculate the quantity of sodium, in mg, per Lozenge: Result = 2.5 C/N = measured quantity of sodium in the Sample solution (g/mL) N = number of Lozenges taken to prepare the Sample solution prepared as directed in the Assay Acceptance criteria: NMT 115.0% of the declared amount SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Analysis: The acid consumed by the minimum single dose recommended in the labeling is NLT 5 mEq of acid and NLT the number of mEq calculated: Result = F (Ta C) F Ta C = acceptable lower limits of the required acidneutralizing capacity, 0.9 = theoretical acid-neutralizing capacity, in mEq of CaCO3 (0.02) = quantity of CaCO3 in the sample tested (mg), based on the labeled quantity Meet the requirements C
IDENTIFICATION IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 6 N hydrochloric acid to a Lozenge produces effervescence, and the resulting solution, after being boiled to expel carbon dioxide and then neutralized with 6 N ammonium hydroxide, meets the requirements of the tests. ASSAY PROCEDURE [NOTEThe Standard solutions and the Sample solution may be modified, if necessary, to obtain solutions, of suitable concentrations, adaptable to the linear or working range of the instrument.] Solution A: Transfer 10 g of potassium chloride and 20 g of lanthanum chloride to a 2000-mL volumetric flask. Add about 1000 mL of water and 40 mL of hydrochloric acid, mix, and allow to cool. Dilute with water to volume, and mix. Solution B: 1 N hydrochloric acid Standard stock solution: 250 mg of chelometric standard calcium carbonate, previously dried at 110 for 2 h and then cooled in a desiccator, in a 500-mL volumetric flask. Add about 100 mL of water and 12 mL of Solution B, swirl to dissolve the calcium carbonate, and allow to cool. Dilute with water to volume. [NOTEThis stock solution contains about 500 g/mL of calcium carbonate.] Standard solution A: 10, 15, and 20 g/mL of calcium carbonate from Standard stock solution in Solution A Standard solution B: 15 g/mL of calcium carbonate from Standard stock solution in Solution A Standard solution C: 20 g/mL calcium carbonate from Standard stock solution in Solution A Sample stock solution: Equivalent to 3000 mg of calcium carbonate, from powdered lozenges, to a 1000-mL volumetric flask, add 100 mL of Solution B and 300 mL of water, and sonicate to dissolve the powder. Dilute with water to volume. Sample solution: Dilute 5.0 mL of Sample stock solution with Solution A to 1000 mL. Blank: Solution A Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Source: Nitrous oxideacetylene Lamp: Calcium hollow-cathode Analytical wavelength: 422.7 nm Analysis Samples: Standard solution A, Standard solution B, and Standard solution C Plot the absorbances of the Standard solutions versus their concentrations of calcium carbonate, in g/mL, by drawing a straight line best fitting the three plotted points. From the graph, determine the concentration, C, in g/mL, of calcium carbonate in the Sample solution. Calculate the percentage of label claim of CaCO3 per Lozenge: Result = (C/CU) 100 = measured concentration of calcium carbonate in the Sample solution (g/mL) = nominal concentration of the Sample solution CU (g/mL) Acceptance criteria: 90.0%110.0% OTHER COMPONENTS SODIUM CONTENT (if so labeled) [NOTEThe Standard solutions and the Sample solution may be modified, if necessary, to obtain solutions of suitable concentrations adaptable to the linear or working range of the instrument.] C
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905:
Calcium Carbonate Oral Suspension

(Comment on this Monograph)id=m11440=Calcium Carbonate Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Calcium Carbonate Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3).
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IDENTIFICATION IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of acetic acid to it produces effervescence, and the resulting solution, after being boiled, meets the requirements. ASSAY PROCEDURE Sample solution: Equivalent to 1 g of calcium carbonate from Oral Suspension, previously shaken in its original container, in a beaker with the aid of 25 mL of water, and add 20 mL of 1 N hydrochloric acid. Heat on a steam bath for 30 min, allow to cool, transfer with the aid of water to a 100-mL volumetric flask, dilute with water to volume, mix, and filter. Transfer 20.0 mL of the filtrate to a suitable container, dilute with water to 100 mL, and add 15 mL of 1 N sodium hydroxide, 5 mL of triethanolamine, and 100 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS until the solution is deep blue. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. Acceptance criteria: 90.0%110.0% IMPURITIES Inorganic Impurities LIMIT OF FLUORIDE [NOTEPrepare and store all solutions in plastic containers.] Solution A: 294 mg/mL of sodium citrate dihydrate Standard stock solution: 1.1052 mg/mL of USP Sodium Fluoride RS Standard solution: Combine 20.0 mL of Standard stock solution with 50.0 mL of Solution A, and dilute to 100 mL with water. Sample solution Transfer an equivalent to 2.0 g of calcium carbonate, to a beaker containing a plastic-coated stirring bar, add 20 mL of water and 2.0 mL of hydrochloric acid, and stir until dissolved. Add 50.0 mL of Solution A and sufficient water to make 100.0 mL. Electrode system: Use a fluoride-specific, ion-indicating electrode and a silversilver chloride reference electrode connected to a pH meter capable of measuring potentials with a minimum reproducibility of 0.2 mV (see pH 791). Analysis Samples: Standard solution and Sample solution Standard response line: Transfer 50.0 mL of Solution A and 2.0 mL of hydrochloric acid to a beaker, and add water to make 100 mL. Add a plastic-coated stirring bar, insert the electrodes into the solution, stir for 15 min, and read the potential, in mV. Continue stirring, and at 5-min intervals add 100, 100, 300, and 500 L of Standard solution, reading the potential 5 min after each addition. Plot the logarithms of the cumulative fluoride ion concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus potential, in mV. Rinse and dry the electrodes, insert them into the Sample solution, stir for 5 min, and read the potential, in mV. From the measured potential and the Standard response line determine the concentration, C (in g/mL) of fluoride ion in the Sample solution. Calculate the concentration, in ppm, of fluoride in the Suspension taken: Result = (C/CU) x F1 = measured concentration of fluoride ion in the Sample solution (g/mL) = concentration of Calcium Carbonate in the CU Sample solution (mg/mL) = unit conversion factor, 1 ppm-mg/g F1 Acceptance criteria: NMT 50 ppm ARSENIC, Method I 211: Slowly dissolve equivalent to 1.0 g from Oral Suspension, previously shaken in its original C

container, in 15 mL of hydrochloric acid, and dilute with water to 55 mL: the resulting solution meets the requirements of the test, the addition of 20 mL of 7 N sulfuric acid specified under Analysis being omitted: NMT 3 ppm. LEAD 251: Mix equivalent to 1.0 g from Oral Suspension, previously shaken in its original container, with 5 mL of water. Slowly add 8 mL of 3 N hydrochloric acid, evaporate on a steam bath to dryness, and dissolve the residue in 5 mL of water: NMT 3 ppm. HEAVY METALS 231: Mix equivalent to 1.0 g from Oral Suspension, previously shaken in its original container, with 5 mL of water. Slowly add 8 mL of 3 N hydrochloric acid, and evaporate on a steam bath to dryness. Dissolve the residue in 20 mL of water, filter, and add water to the filtrate to make 25 mL: NMT 20 ppm. SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62 and TESTS FOR SPECIFIED MICROORGANISMS 62: Its total aerobic microbial count: NMT 100 cfu/mL, and it meets the requirements of the test for absence of Escherichia coli and Pseudomonas aeruginosa. PH 791: 7.58.7 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing. USP REFERENCE STANDARDS 11 USP Sodium Fluoride RS
Calcium Carbonate Tablets

(Comment on this Monograph)id=m11450=Calcium Carbonate Tablets=Ca-Chl-Monos.pdf) DEFINITION Calcium Carbonate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3). For Tablets labeled for any indication other than, or in addition to, antacid use the Tablets contain NLT 90.0% and NMT 115.0% of the labeled amount of calcium carbonate. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 6 N acetic acid to the Tablets produces effervescence, and the resulting solution, after being boiled to expel carbon dioxide and neutralized with 6 N ammonium hydroxide, meets the requirements of the tests. ASSAY PROCEDURE Sample: Equivalent to 200 mg of calcium carbonate, from powdered Tablets (NLT 20), in a suitable crucible Analysis: Ignite to constant weight. Cool the crucible, add 10 mL of water, and dissolve the residue by adding sufficient 3 N hydrochloric acid, dropwise, to achieve complete solution. Transfer the solution completely to a suitable container, dilute with water to 150 mL, and add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue. Titrate with 0.05 M edetate disodium VS until the solution is deep blue. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. Acceptance criteria: 90.0%110.0% of the labeled amount of CaCO3; for Tablets labeled for any indication other than, or in addition to, antacid use, 90.0%115.0% of the labeled amount of CaCO3
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SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Analysis: Where Tablets are labeled for antacid use, NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and NLT the number of mEq calculated: Result = (Ta C) Ta C = theoretical acid-neutralizing capacity of CaCO3 (mEq), 0.02 = quantity of CaCO3 in the sample tested (mg), based on the labeled quantity
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate whether it is for use as an antacid, or as a dietary supplement, or both.
Calcium Carbonate and Magnesia Tablets

(Comment on this Monograph)id=m11474=Calcium Carbonate and Magnesia Tablets=Ca-Chl-Monos.pdf) (Current titlenot to change until February 1, 2010) Monograph title changeto become official February 1, 2010 See Calcium Carbonate and Magnesia Chewable Tablets DEFINITION Calcium Carbonate and Magnesia Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3) and NLT 90.0% and NMT 115.0% of the labeled amount of magnesium hydroxide [Mg(OH)2]. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 3 N hydrochloric acid to the Tablets produces effervescence, and the resulting solution, after being boiled to expel carbon dioxide and neutralized with 6 N ammonium hydroxide, meets the requirements of the tests. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Heat 2 Tablets in 20 mL of 1 N sulfuric acid. Analysis: Cool, add 20 mL of alcohol, mix, and allow to stand for 30 min. Filter this solution, and add 2 mL of 1 N hydrochloric acid to the filtrate. Acceptance criteria: Meet the requirements ASSAY PROCEDURE 1: CALCIUM CARBONATE Sample solution: Transfer an equivalent to 400 mg of calcium carbonate, from powdered Tablets (NLT 20), in 25 mL of water, and add 40 mL of 1 N hydrochloric acid. Heat on a steam bath for 30 min, allow to cool, transfer to a 100mL volumetric flask with the aid of water, dilute with water to volume, mix, and filter. Transfer 20.0 mL of the filtrate to a suitable container, dilute with water to 100 mL, and add 30 mL of 1 N sodium hydroxide, 5 mL of triethanolamine, and 100 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS until the solution is deep blue in color. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. PROCEDURE 2: MAGNESIUM HYDROXIDE Sample solution: Equivalent to 120 mg of calcium carbonate and magnesium hydroxide combined, from the portion of the filtrate remaining from the Assay for Calcium Carbonate, to a suitable container, dilute with water to 100 mL, and add 10 mL of ammoniaammonium chloride buffer TS, 5 mL of triethanolamine, and 0.3 mL of eriochrome black TS. Analysis: Titrate with 0.05 M edetate disodium VS to a blue endpoint. The volume, in mL, of 0.05 M edetate disodium consumed, less the volume of 0.05 M edetate disodium corresponding to the content of calcium carbonate in the volume, in mL, of the filtrate taken, represents the volume, in mL, of 0.05 M edetate disodium equivalent to the quantity of magnesium hydroxide present. Each mL of 0.05 M edetate disodium is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% of the labeled amount of CaCO3; 90.0%115.0% of the labeled amount of Mg(OH)2
PERFORMANCE TESTS DISSOLUTION 711 [NOTEFor Tablets labeled for any indication other than, or in addition to, antacid use.] Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 75 rpm Time: 30 min Analysis: Determine the amount of CaCO3 dissolved by employing the following method. Solution A: 50 mg/mL of lanthanum chloride in 0.1 N hydrochloric acid Blank: Solution A and 0.1 N hydrochloric acid (1:9) Standard stock solution: 100 g/mL of calcium carbonate in 0.1 N hydrochloric acid Standard solution A: Dilute 3 mL of Standard stock solution and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 mL. Standard solution B: Dilute 4 mL of Standard stock solution and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 mL. Standard solution C: Dilute 5 mL of Standard stock solution and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 mL. Standard solution D: Dilute 6 mL of Standard stock solution and 10 mL of Solution A with 0.1 N hydrochloric acid to 100 mL. Sample solution: Sample per Dissolution 711. Filter a portion of the solution under test. Pipet a volume of the filtrate, estimated to contain 1 mg of calcium, into a 250-mL volumetric flask, add 25.0 mL of Solution A, and dilute with 0.1 N hydrochloric acid to volume. Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Lamp: Calcium hollow-cathode Flame: Airacetylene Analytical wavelength: 422.8 nm Analysis Samples: Standard solution A, Standard solution B, Standard solution C, Standard solution D, Sample solution, and Blank Concomitantly determine the absorbances of the Standard solutions and the Sample solution, against the Blank. Construct a standard curve by plotting absorbances versus calcium concentrations of the Standard solutions, then from it obtain the concentration, Cs, in g/mL of calcium, of the Sample solution. Calculate the percentage of CaCO3 dissolved: Result = (Mr/Ar) (CS/CU) 100 = molecular weight of calcium carbonate, 100.09 = atomic weight of calcium, 40.08 = measured concentration of calcium in the Sample solution (g/mL) CU = nominal concentration of the Sample solution (g/mL) Tolerances: NLT 75% (Q) of the labeled amount of CaCO3 is dissolved. Mr Ar CS
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SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Analysis: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and NLT the number of mEq calculated: Result = Fa (Ta M) + Fb (Tb C) Fa Ta M Fb Tb C = monograph correction factor for Mg(OH)2, 0.8 = theoretical acid-neutralizing capacity of Mg(OH)2 (mEq), 0.0343 = quantity of Mg(OH)2 in the specimen tested (mg), based on the labeled quantities = monograph correction factor for CaCO3, 0.9 = theoretical acid-neutralizing capacities of CaCO3 (mEq), 0.02 = quantity of CaCO3 in the specimen tested (mg), based on the labeled quantities

Analysis: Titrate with 0.05 M edetate disodium VS until the solution is deep blue in color. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. PROCEDURE 2: MAGNESIUM HYDROXIDE Sample solution: Equivalent to 120 mg of calcium carbonate and magnesium hydroxide combined, from the portion of the filtrate remaining from the Assay for Calcium Carbonate, to a suitable container, dilute with water to 100 mL, and add 10 mL of ammoniaammonium chloride buffer TS, 5 mL of triethanolamine, and 0.3 mL of eriochrome black TS. Analysis: Titrate with 0.05 M edetate disodium VS to a blue endpoint. The volume, in mL, of 0.05 M edetate disodium consumed, less the volume of 0.05 M edetate disodium corresponding to the content of calcium carbonate in the volume, in mL, of the filtrate taken, represents the volume, in mL, of 0.05 M edetate disodium equivalent to the quantity of magnesium hydroxide present. Each mL of 0.05 M edetate disodium is equivalent to 2.916 mg of Mg(OH)2. Acceptance criteria: 90.0%110.0% of the labeled amount of CaCO3; 90.0%115.0% of the labeled amount of Mg(OH)2 SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Analysis: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and NLT the number of mEq calculated: Result = Fa (Ta M) + Fb (Tb C) Fa Ta M Fb Tb C = monograph correction factor for Mg(OH)2, 0.8 = theoretical acid-neutralizing capacity of Mg(OH)2 (mEq), 0.0343 = quantity of Mg(OH)2 in the specimen tested (mg), based on the labeled quantities = monograph correction factor for CaCO3, 0.9 = theoretical acid-neutralizing capacities of CaCO3 (mEq), 0.02 = quantity of CaCO3 in the specimen tested (mg), based on the labeled quantities
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to calcium carbonate and to magnesia ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Tablets to indicate that they must be chewed before being swallowed.
Calcium Carbonate and Magnesia Chewable Tablets

(Comment on this Monograph)id=m2520=Calcium Carbonate and Magnesia Chewable Tablets=Ca-Chl-Monos.pdf) (Monograph under this new titleto become official February 1, 2010) (Current monograph title is Calcium Carbonate and Magnesia Tablets) DEFINITION Calcium Carbonate and Magnesia Chewable Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3) and NLT 90.0% and NMT 115.0% of the labeled amount of magnesium hydroxide [Mg(OH)2]. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 3 N hydrochloric acid to the Chewable Tablets produces effervescence, and the resulting solution, after being boiled to expel carbon dioxide and neutralized with 6 N ammonium hydroxide, meets the requirements of the tests. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Heat 2 Chewable Tablets in 20 mL of 1 N sulfuric acid. Analysis: Cool, add 20 mL of alcohol, mix, and allow to stand for 30 min. Filter this solution, and add 2 mL of 1 N hydrochloric acid to the filtrate. Acceptance criteria: Meet the requirements ASSAY PROCEDURE 1: CALCIUM CARBONATE Sample solution: Transfer an equivalent to 400 mg of calcium carbonate, from powdered Chewable Tablets (NLT 20), in 25 mL of water, and add 40 mL of 1 N hydrochloric acid. Heat on a steam bath for 30 min, allow to cool, transfer to a 100-mL volumetric flask with the aid of water, dilute with water to volume, mix, and filter. Transfer 20.0 mL of the filtrate to a suitable container, dilute with water to 100 mL, and add 30 mL of 1 N sodium hydroxide, 5 mL of triethanolamine, and 100 mg of hydroxy naphthol blue.
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to calcium carbonate and to magnesia ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label the Chewable Tablets to indicate that they must be chewed before being swallowed.
Calcium Carbonate, Magnesia, and Simethicone Tablets

(Comment on this Monograph)id=m11476=Calcium Carbonate, Magnesia, and Simethicone Tablets=Ca-Chl-Monos.pdf) (Current titlenot to change until February 1, 2010) Monograph title changeto become official February 1, 2010 See Calcium Carbonate, Magnesia, and Simethicone Chewable Tablets DEFINITION Calcium Carbonate, Magnesia, and Simethicone Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of calcium carbonate (CaCO3) and magnesium hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane [-(CH3)2SiO-]n that is NLT 85.0% and NMT 115.0% of the labeled amount of simethicone.
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volumetric flask, add 1.0 mL of Solution A, and dilute with 4methyl-2-pentanone to volume. Blank solution: 1.0 mL Solution A diluted with 4-methyl-2pentanone to 50 mL Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Lamp: Silicon hollow-cathode lamp Flame: Nitrous oxcideacetylene Analytical wavelength: 251.6 nm Analysis Samples: Standard solution, Sample solution, and Blank solution Calculate the percent label claim of polydimethylsiloxane: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the the Standard solution = concentration of USP Polydimethylsiloxane RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU CALCIUM CARBONATE Solution A: Transfer 26.8 g of lanthanum chloride to a 200-mL volumetric flask, add 100 mL of water, and carefully add 50 mL of hydrochloric acid, mix and allow to cool. Dilute with water to volume. Solution B: Prepare as directed in the Assay for Polydimethylsiloxane. Standard stock solution A: Transfer 499.5 mg of primary standard calcium carbonate to a 200-mL volumetric flask, and add 10 mL of water. Carefully add 5 mL of Solution B, and swirl to dissolve the calcium carbonate. Dilute with water to volume. [NOTEThis solution contains 1000 g/mL of calcium (Ca).] Standard stock solution B: 1.000 g of magnesium metal to a 1000-mL volumetric flask containing 10 mL of water, slowly add 10 mL of hydrochloric acid, and swirl to dissolve the metal. Dilute with water to volume. [NOTEThis solution contains 1000 g/mL of magnesium (Mg).] Standard solution A: To a 250-mL volumetric flask, add 10.0 mL of Standard stock solution A and 5.0 mL of Standard stock solution B, and dilute with water to volume. [NOTEThis solution contains 40 g/mL of calcium (Ca) and 20 g/mL of magnesium (Mg).] Standard solution: On the day of use, transfer 4.0 mL of Standard solution A to a 100-mL volumetric flask containing 2.0 mL of Solution A, and dilute with water to volume. [NOTEThis solution contains 1.6 g/mL of calcium (Ca) and 0.8 g/mL of magnesium (Mg).] Sample stock solution: Transfer a volume of the aqueous layer retained from the Sample solution in the Assay for Polydimethylsiloxane, equivalent to about 28 mg of calcium carbonate, to a 200-mL volumetric flask, and dilute with water to volume. Sample solution: Transfer 3.0 mL of Sample stock solution to a 100-mL volumetric flask containing 2.0 mL of Solution A to dilute with water to volume. Blank solution: Transfer 5.0 mL of Solution A to a 250-mL volumetric flask, and dilute with water to volume. Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Lamp: Calcium hollow-cathode Flame: Nitrous oxideacetylene Analytical wavelength: 422.7 nm Analysis Samples: Standard solution, Sample solution, and Blank solution AU AS CS
IDENTIFICATION A. INFRARED ABSORPTION 197S Standard solution: Transfer 33 mg of USP Polydimethylsiloxane RS, to a suitable round, narrow-mouth, screw-capped, 120-mL bottle, add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 20.0 mL of toluene, close the bottle securely with a cap having an inert liner, and shake for 30 min on a reciprocating shaker (e.g., about 200 oscillations/min and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator, and allow to separate. Remove the upper, organic layer to a screw-capped, centrifuge tube containing about 2 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. Sample solution: Transfer an equivalent to 33 mg of simethicone from powdered Tablets (NLT 20 Tablets), to a suitable round, narrow-mouth, screw-capped, 120-mL bottle. Add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 20.0 mL of toluene, close the bottle securely with a cap having an inert liner, and shake for 30 min on a reciprocating shaker (e.g., about 200 oscillations/min and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator, and allow to separate. Remove the upper, organic layer to a screw-capped, centrifuge tube containing about 2 g of anhydrous sodium sulfate. Close the tube with a screwcap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. Blank: 10 mL of toluene with about 1 g of anhydrous sodium sulfate, centrifuged to obtain a clear supernatant Cell: 0.5 mm Analysis: Concomitantly determine the absorbances of the Sample solution and Standard solution at a wavelength of maximum absorbance at about 7.9 m (1265.8 cm1). B. IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 1 N hydrochloric acid to a Tablet produces effervescence, and the resulting solution, after having been filtered, meets the requirements. C. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Heat 2 Tablets in 20 mL of 1 N sulfuric acid. Analysis: Cool, add 20 mL of alcohol, mix, and allow to stand for 30 min. Filter this solution, and to the filtrate, add 2 mL of 1 N hydrochloric acid: this solution meets the requirements. ASSAY POLYDIMETHYLSILOXANE Solution A: 12.5 mg/mL of saccharin in 4-methyl-2pentanone Solution B: 2.4 M hydrochloric acid Standard stock solution: 1 mg/mL of USP Polydimethylsiloxane RS in 4-methyl-2-pentanone Standard solution: Transfer 20.0 mL of Standard stock solution and 5.0 mL of Solution A into a 250-mL volumetric flask. Dilute to volume with 4-methyl-2-pentanone. [NOTEPrepare the Standard solution on the day of use only. This solution contains about 0.08 mg/mL of USP Polydimethylsiloxane RS.] Sample solution: Transfer an equivalent to 20 mg of polydimethylsiloxane from powdered Tablets (NLT 20 Tablets), to a 125-mL separator. Cautiously add 50.0 mL of Solution B, and swirl until the reaction subsides. Insert the stopper, and mix. Carefully release the pressure, add 50.0 mL of 4-methyl-2-pentanone, and mix for 10 min. Allow the layers to separate, and drain the aqueous layer into a suitable stoppered container. [NOTERetain this aqueous layer for use in preparing the Sample solution in the Assay for Calcium carbonate and Magnesium hydroxide and for the Sample solution in the test for Sodium content.] Pass the organic layer through a filter containing 50 g of anhydrous sodium sulfate. Transfer 10.0 mL of the filtrate to a 50-mL
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Calculate the percent label claim of CaCO3: Result = (Mr/Ar) (CS/CU) (AU/AS) 100 Mr Ar CS CU AU AS = molecular weight of calcium carbonate, 100.09 = atomic weight of calcium, 40.08 = concentration of calcium in the Standard solution (g/mL) = nominal concentration in the Sample solution (g/mL) = absorbance of the Sample solution = absorbance of the the Standard solution

Mode: Atomic absorption spectrometer Lamp: Sodium hollow-cathode Flame: Airacetylene Analytical wavelength: 589.0 nm Analysis Samples: Standard solution, Sample solution, and Blank solution Calculate the mg of sodium in each Tablet: (5C/6) (A/W) (AU/AS) C A W AU AS = concentration of sodium from the Standard solution (g/mL) = average weight of each Tablet (mg) = weight of the portion of Tablets taken to prepare the Sample solution in the Assay for Polydimethylsiloxane (mg) = absorbance of the Sample solution = absorbance of the Standard solution
MAGNESIUM HYDROXIDE Solution A, Solution B, Standard stock solution A, Standard stock solution B, Standard solution A, Standard solution B, Sample stock solution, Sample solution, and Blank solution: Proceed as directed in Assay for Calcium Carbonate. Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Lamp: Magnesium hollow-cathode Flame: Nitrous oxideacetylene Analytical wavelength: 285.2 nm Analysis Samples: Standard solution, Sample solution, and Blank solution Calculate the percent label claim of Mg(OH)2: Result = (AU/AS) (CS/CU) (Mr/Ar) 100
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to calcium carbonate and to magnesium hydroxide SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that the Tablets are to be chewed before swallowing. Label the Tablets to state the sodium content, in mg/Tablet, if it is greater than 5 mg/Tablet. USP REFERENCE STANDARDS 11 USP Polydimethylsiloxane RS
= absorbance of the Sample solution = absorbance of the the Standard solution = concentration of calcium of the Standard solution (g/mL) = concentration of the Sample solution (g/mL) CU = molecular weight of magnesium hydroxide, Mr 58.34 = atomic weight of magnesium, 24.305 Ar Acceptance criteria: 90.0%110.0% of the labeled amounts of CaCO3 and Mg(OH)2; and an amount of polydimethylsiloxane [(CH3)2SiO]n, 85.0%115.0% of the labeled amount of simethicone AU AS CS OTHER COMPONENTS SODIUM CONTENT (if so labeled): Each Tablet contains NMT the number of mg of sodium stated on the label. Solution A: Prepare as directed in the Assay for Calcium Carbonate. Solution B: Prepare as directed in the Assay for Polydimethylsiloxane. Standard stock solution: 2.542 mg/mL of sodium chloride, previously dried at 105 for 2 h, in water. Transfer 5.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 4.0 mL of this solution to a second 100-mL volumetric flask containing 6.0 mL of Solution B and 2.0 mL of Solution A, and dilute with water to volume. This solution contains 2.0 g of sodium/mL. Sample solution: Transfer 3.0 mL of the aqueous layer retained from the solution of the Sample solution in the Assay for Polydimethylsiloxane to a 50-mL volumetric flask containing 1.0 mL of Solution A, and dilute with water to volume. Blank solution: 15.0 mL Solution B and 5.0 mL Solution A diluted with water to 250 mL Spectrometric conditions (See Spectrometry and Light-Scattering 851.)
Calcium Carbonate, Magnesia, and Simethicone Chewable Tablets

(Comment on this Monograph)id=m11476=Calcium Carbonate, Magnesia, and Simethicone Chewable Tablets=Ca-ChlMonos.pdf) Monograph title changeto become official February 1, 2010 (Current titlenot to change until February 1, 2010) See Calcium Carbonate, Magnesia, and Simethicone Tablets DEFINITION Calcium Carbonate, Magnesia, and Simethicone Chewable Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of calcium carbonate (CaCO3) and magnesium hydroxide [Mg(OH)2], and an amount of polydimethylsiloxane [-(CH3)2SiO-]n that is NLT 85.0% and NMT 115.0% of the labeled amount of simethicone. IDENTIFICATION A. INFRARED ABSORPTION 197S Standard solution: Transfer 33 mg of USP Polydimethylsiloxane RS, to a suitable round, narrow-mouth, screw-capped, 120-mL bottle, add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 20.0 mL of toluene, close the bottle securely with a cap having an inert liner, and shake for 30 min, on a reciprocating shaker (e.g., about 200 oscillations/min and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator, and allow to separate.
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Mode: Atomic absorption spectrometer Lamp: Silicon hollow-cathode lamp Flame: Nitrous oxcideacetylene Analytical wavelength: 251.6 nm Analysis Samples: Standard solution, Sample solution, and Blank solution Calculate the percent label claim of polydimethylsiloxane: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the the Standard solution = concentration of USP Polydimethylsiloxane RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU CALCIUM CARBONATE AND MAGNESIUM HYDROXIDE Solution A: Transfer 26.8 g of lanthanum chloride to a 200-mL volumetric flask, add 100 mL of water, and carefully add 50 mL of hydrochloric acid, mix and allow to cool. Dilute with water to volume. Solution B: Prepare as directed in the Assay for Polydimethylsiloxane. Standard stock solution A: Transfer 499.5 mg of primary standard calcium carbonate to a 200-mL volumetric flask, and add 10 mL of water. Carefully add 5 mL of Solution B, and swirl to dissolve the calcium carbonate. Dilute with water to volume. [NOTEThis solution contains 1000 g/mL of calcium (Ca).] Standard stock solution B: 1.000 g of magnesium metal to a 1000-mL volumetric flask containing 10 mL of water, slowly add 10 mL of hydrochloric acid, and swirl to dissolve the metal. Dilute with water to volume. [NOTEThis solution contains 1000 g/mL of magnesium (Mg).] Standard solution A: To a 250-mL volumetric flask, add 10.0 mL of Standard stock solution A and 5.0 mL of Standard stock solution B, and dilute with water to volume. [NOTEThis solution contains 40 g/mL of calcium (Ca) and 20 g/mL of magnesium (Mg).] Standard solution: On the day of use, transfer 4.0 mL of Standard solution A to a 100-mL volumetric flask containing 2.0 mL of Solution A, and dilute with water to volume. [NOTEThis solution contains 1.6 g/mL of calcium (Ca) and 0.8 g/mL of magnesium (Mg).] Sample stock solution: Transfer volume of the aqueous layer retained from the Sample solution in the Assay for Polydimethylsiloxane, equivalent to about 28 mg of calcium carbonate, to a 200-mL volumetric flask, and dilute with water to volume. Sample solution: Transfer 3.0 mL of Sample stock solution to a 100-mL volumetric flask containing 2.0 mL of Solution A to dilute with water to volume. Blank solution: Transfer 5.0 mL of Solution A to a 250-mL volumetric flask, and dilute with water to volume. CALCIUM CARBONATE Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Lamp alcium hollow-cathode lamp Flame: nitrous oxide-acetylene flame Analytical wavelength: 422.7 nm Analysis Samples: Standard solution, Sample solution, and Blank solution Calculate the percent label claim, of CaCO3: AU AS CS Result = (AU/AS) (CS/CU) (Mr/Ar) 100
Remove the upper, organic layer to a screw-capped, centrifuge tube containing about 2 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. Sample solution: Transfer an equivalent to 33 mg of simethicone from powdered Chewable Tablets (NLT 20 Chewable Tablets), to a suitable round, narrow-mouth, screw-capped, 120-mL bottle. Add 40 mL of 0.1 N sodium hydroxide, and swirl to disperse. Add 20.0 mL of toluene, close the bottle securely with a cap having an inert liner, and shake for 30 min on a reciprocating shaker (e.g., about 200 oscillations/min and a stroke of 38 2 mm). Transfer the mixture to a 125-mL separator, and allow to separate. Remove the upper, organic layer to a screw-capped, centrifuge tube containing about 2 g of anhydrous sodium sulfate. Close the tube with a screw-cap having an inert liner, agitate vigorously, and centrifuge the mixture until a clear supernatant is obtained. Blank: 10 mL of toluene with about 1 g of anhydrous sodium sulfate, centrifuged to obtain a clear supernatant Cell: 0.5 mm Analysis: Concomitantly determine the absorbances of the Sample solution and Standard solution at a wavelength of maximum absorbance at about 7.9 m (1265.8 cm1). B. IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 1 N hydrochloric acid to a Chewable Tablet produces effervescence, and the resulting solution, after having been filtered, meets the requirements. C. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Heat 2 Chewable Tablets in 20 mL of 1 N sulfuric acid. Analysis: Cool, add 20 mL of alcohol, mix, and allow to stand for 30 min. Filter this solution, and to the filtrate, add 2 mL of 1 N hydrochloric acid: this solution meets the requirements. ASSAY POLYDIMETHYLSILOXANE Solution A: 12.5 mg/mL of saccharin in 4-methyl-2pentanone Solution B: 2.4 M hydrochloric acid Standard stock solution: 1 mg/mL of USP Polydimethylsiloxane RS in 4-methyl-2-pentanone Standard solution: Transfer 20.0 mL of Standard stock solution and 5.0 mL of Solution A into a 250-mL volumetric flask. Dilute with 4-methyl-2-pentanone to volume. [NOTEPrepare the Standard solution on the day of use only. This solution contains about 0.08 mg/mL of USP Polydimethylsiloxane RS.] Sample solution: Transfer an equivalent to 20 mg of polydimethylsiloxane from powdered Chewable Tablets (NLT 20 Chewable Tablets), to a 125-mL separator. Cautiously add 50.0 mL of Solution B, and swirl until the reaction subsides. Insert the stopper, and mix. Carefully release the pressure, add 50.0 mL of 4-methyl-2-pentanone, and mix for 10 min. Allow the layers to separate, and drain the aqueous layer into a suitable stoppered container. [NOTE Retain this aqueous layer for use in preparing the Sample solution in the Assay for Calcium carbonate and Magnesium hydroxide and for the Sample solution in the test for Sodium content.] Pass the organic layer through a filter containing 50 g of anhydrous sodium sulfate. Transfer 10.0 mL of the filtrate to a 50-mL volumetric flask, add 1.0 mL of Solution A, and dilute with 4-methyl-2-pentanone to volume. Blank solution: 1.0 mL Solution A diluted with 4-methyl-2pentanone to 50 mL Spectrometric conditions (See Spectrometry and Light-Scattering 851.)
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AU AS CS CU = absorbance of the Sample solution = absorbance of the the Standard solution = concentration of calcium in the Standard solution (g/mL) = nominal concentration of the Sample solution (g/mL) = molecular weight of calcium carbonate, 100.09 = atomic weight of calcium, 40.08

Calculate the mg of sodium in each Chewable Tablet: Result = (AU/AS) (A/W) (5C/6) AU AS A W C = = = = absorbance of the Sample solution absorbance of the Standard solution average weight of each Chewable Tablet (mg) weight of the portion of Chewable Tablets taken to prepare the Sample solution in the Assay for Polydimethylsiloxane (mg) = concentration of sodium in the Standard solution (g/mL)
Mr Ar MAGNESIUM HYDROXIDE Solution A, Solution B, Standard stock solution A, Standard stock solution B, Standard solution A, Standard solution B, Sample stock solution, Sample solution, and Blank solution: Proceed as directed in Assay for Calcium carbonate. Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Lamp: Magnesium hollow-cathode Flame: Nitrous oxideacetylene Analytical wavelength: 285.2 nm Analysis Samples: Standard solution, Sample solution, and Blank solution Calculate the percent label claim of [Mg(OH)2]: Result = (Mr/Ar) (CS/CU) (AU/AS) 100
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation with respect to calcium carbonate and to magnesium hydroxide SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that the Chewable Tablets are to be chewed before swallowing. Label the Chewable Tablets to state the sodium content, in mg/Chewable Tablet, if it is greater than 5 mg/Chewable Tablet. USP REFERENCE STANDARDS 11 USP Polydimethylsiloxane RS
= molecular weight of magnesium hydroxide, 58.34 = atomic weight of magnesium, 24.305 Ar = concentration of calcium in the Standard CS solution (g/mL) = concentration of the Sample solution (g/mL) CU AU = absorbance of the Sample solution AS = absorbance of the the Standard solution Acceptance criteria: 90.0%110.0% of the labeled amounts of CaCO3 and Mg(OH)2; and an amount of polydimethylsiloxane [-(CH3)2SiO-]n, 85.0%115.0% of the labeled amount of simethicone OTHER COMPONENTS SODIUM CONTENT (if so labeled): Each Chewable Tablet contains NMT the number of mg of sodium stated on the label. Solution A: Prepare as directed in the Assay for Calcium carbonate and Magnesium hydroxide. Solution B: Prepare as directed in the Assay for Polydimethylsiloxane. Standard stock solution: 2.542 mg/mL of sodium chloride, previously dried at 105 for 2 h, in water. Transfer 5.0 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Transfer 4.0 mL of this solution to a second 100-mL volumetric flask containing 6.0 mL of Solution B and 2.0 mL of Solution A, and dilute with water to volume. This solution contains 2.0 g of sodium/mL. Sample solution: Transfer 3.0 mL of the aqueous layer retained from the solution of the Sample solution in the Assay for Polydimethylsiloxane to a 50-mL volumetric flask containing 1.0 mL of Solution A, and dilute with water to volume. Blank solution: 15.0 mL Solution B and 5.0 mL Solution A diluted with water to 250 mL Spectrometric conditions (See Spectrometry and Light-Scattering 851.) Mode: Atomic absorption spectrometer Lamp: Sodium hollow-cathode Flame: AirAcetylene Analytical wavelength: 589.0 nm Analysis Samples: Standard solution, Sample solution, and Blank solution
Mr
Calcium and Magnesium Carbonates Oral Suspension

(Comment on this Monograph)id=m11478=Calcium and Magnesium Carbonates Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Calcium and Magnesium Carbonates Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3) and NLT 85.0% and NMT 115.0% of the labeled amount of MgCO3. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 3 N hydrochloric acid to a quantity of Oral Suspension, equivalent to 500 mg of calcium carbonate, produces effervescence, and the resulting solution, after having been filtered, meets the requirements. B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Heat a quantity of Oral Suspension, equivalent to 800 mg of magnesium carbonate, with 20 mL of 1 N sulfuric acid. Analysis: Cool, add 20 mL of alcohol, mix, and allow to stand for 30 min. Filter this solution, and add 2 mL of 1 N hydrochloric acid to the filtrate. Acceptance criteria: Meets the requirements ASSAY CALCIUM CARBONATE Sample solution: Transfer the equivalent to 400 mg of calcium carbonate from Oral Suspension, previously well shaken in its original container and free of air bubbles, to a beaker, with the aid of 20 mL of water, and add 10 mL of 1 N hydrochloric acid. Heat on a steam bath for 30 min, allow to cool, transfer with the aid of water to a 100-mL volumetric flask, dilute with water to volume, and filter. Analysis: Transfer 20.0 mL of Sample solution to a suitable container, dilute with water to 100.0 mL, add 15 mL of 1 N sodium hydroxide, 5 mL of triethanolamine, and 100 mg of
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B. IDENTIFICATION TESTSGENERAL, Magnesium 191 Sample solution: Heat 2 Tablets in 20 mL of 1 N sulfuric acid. Analysis: Cool, add 20 mL of alcohol, mix, and allow to stand for 30 min. Filter this solution, and add 2 mL of 1 N hydrochloric acid to the filtrate. Acceptance criteria: The filtrate meets the requirements. ASSAY CALCIUM CARBONATE Sample solution: Transfer the equivalent of 400 mg of calcium carbonate, from NLT 20 powdered Tablets, to a beaker with the aid of 25 mL of water, and add 10 mL of 1 N hydrochloric acid. Heat on a steam bath for 30 min, allow to cool, transfer with the aid of water to a 100-mL volumetric flask, dilute with water to volume, mix, and filter. Analysis: Transfer 20.0 mL of Sample solution to a suitable container, dilute with water to 100 mL, add 15 mL of 1 N sodium hydroxide, 5 mL of triethanolamine, and 100 mg of hydroxy naphthol blue. Titrate with 0.05 M edetate disodium VS until the solution is deep blue. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. Acceptance criteria: 90.0%110.0% for CaCO3 MAGNESIUM CARBONATE Sample solution: Equivalent of 120 mg of calcium carbonate and magnesium carbonate from the Sample solution in the Assay for Calcium carbonate. Dilute with water to 100 mL, and add 10 mL of ammoniaammonium chloride buffer TS, 5 mL of triethanolamine, and 0.3 mL of eriochrome black TS. Analysis: Titrate with 0.05 M edetate disodium VS to a blue endpoint. From the volume of 0.05 M edetate disodium consumed, deduct the volume of 0.05 M edetate disodium used in Assay. The difference is the volume of 0.05 M edetate disodium equivalent to the amount of magnesium carbonate present. Each mL of 0.05 M edetate disodium is equivalent to 4.216 mg of MgCO3. Acceptance criteria: 85.0%115.0% for MgCO3 PERFORMANCE TESTS DISINTEGRATION 701 Immersion fluid: Substitute simulated gastric fluid TS for water. Time: 10 min; except that where Tablets are labeled as gelatin-coated, the time is 30 min UNIFORMITY OF DOSAGE UNITS, Weight Variation 905: Meet the requirements for CaCO3 and MgCO3 SPECIFIC TESTS ACID-NEUTRALIZING CAPACITY 301 Analysis: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and not less than the number of mEq calculated. Calculate mEq: Result = [0.8 (ANC1M)] + [0.9 (ANC2C)] ANC1 M ANC2 C = theoretical acid-neutralizing capacity of MgCO3, 0.024 mEq = quantity of MgCO3 in the specimen tested, based on the labeled quantities (mg) = theoretical acid-neutralizing capacity of CaCO3, 0.02 mEq = quantity of CaCO3 in the specimen tested, based on the labeled quantities (mg)
hydroxynaphthol blue trituration. Titrate with 0.05 M edetate disodium VS until the solution is deep blue. Each mL of 0.05 M edetate disodium is equivalent to 5.004 mg of CaCO3. MAGNESIUM CARBONATE Sample solution: Equivalent to 120 mg of calcium carbonate and magnesium carbonate from the Sample solution in the Assay for Calcium carbonate, dilute with water to 100 mL, and add 10 mL of ammonia-ammonium chloride buffer TS, 5 mL of triethanolamine, and 0.3 mL of eriochrome black TS. Analysis: Titrate with 0.05 M edetate disodium VS to a blue endpoint. From the volume of 0.05 M edetate disodium consumed, subtract the volume of 0.05 M edetate disodium used in Assay. The difference is the volume of 0.05 M edetate disodium equivalent to the amount of magnesium carbonate present. Each mL of 0.05 M edetate disodium is equivalent to 4.216 mg of MgCO3. Acceptance criteria: 90.0%110.0% of the labeled amount of CaCO3; 85.0%115.0% of the labeled amount of MgCO3 PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: Total aerobic microbial count: NMT 100 cfu/mL, and it meets the requirements of the tests for absence of Escherichia coli and Pseudomonas aeruginosa PH 791: 7.08.6 ACID-NEUTRALIZING CAPACITY 301: NLT 5 mEq of acid is consumed by the minimum single dose recommended in the labeling, and NLT the number of mEq calculated: Result = 0.8 (0.024 M) + 0.9 (0.02 C) 0.024 0.02 M C = theoretical acid-neutralizing capacity MgCO3 (mEq) = theoretical acid-neutralizing capacities CaCO3 (mEq) = quantity of MgCO3 in the specimen tested, based on the labeled quantities (mg) = quantity of CaCO3 in the specimen tested, based on the labeled quantities (mg)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid freezing.
Calcium and Magnesium Carbonates Tablets

(Comment on this Monograph)id=m11480=Calcium and Magnesium Carbonates Tablets=Ca-Chl-Monos.pdf) DEFINITION Calcium and Magnesium Carbonates Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of calcium carbonate (CaCO3) and NLT 85.0% and NMT 115.0% of the labeled amount of magnesium carbonate (MgCO3). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: The addition of 1 N hydrochloric acid to 1 Tablet produces effervescence, and the resulting solution, after having been filtered, meets the requirements of the tests.
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Tablets that are gelatin-coated are so labeled.

volume. Carefully heat over a free flame to dryness, and continue heating to complete decomposition and volatilization of ammonium salts. Finally ignite the residue to constant weight. Acceptance criteria: The weight of the residue is NMT 5 mg (1.0%). SPECIFIC TESTS PH 791: 4.59.2, in 50 mg/mL solution 147.01 110.98 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where Calcium Chloride is intended for use in hemodialysis, it is so labeled.
Calcium Chloride
(Comment on this Monograph)id=m11500=Calcium Chloride=Ca-Chl-Monos.pdf) CaCl2 2H2O Calcium chloride dihydrate [10035-04-8]. Anhydrous [10043-52-4].
DEFINITION Calcium Chloride contains an amount of CaCl2 equivalent to NLT 99.0% and NMT 107.0% of CaCl2 2H2O. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: mg/mL B. IDENTIFICATION TESTSGENERAL, Chloride 191: mg/mL 100 100
Calcium Chloride Injection

(Comment on this Monograph)id=m11510=Calcium Chloride Injection=Ca-Chl-Monos.pdf) DEFINITION Calcium Chloride Injection is a sterile solution of Calcium Chloride in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of CaCl2 2H2O. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191 B. IDENTIFICATION TESTSGENERAL, Chloride 191 ASSAY PROCEDURE Sample solution: 1 g of calcium chloride equivalent Injection in a 250-mL volumetric flask, add 5 mL of 3 N hydrochloric acid, and dilute with water to volume. Pipet 50 mL of the resulting solution into a suitable container, add 100 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS until the solution is deep blue. Each mL of 0.05 M edetate disodium is equivalent to 7.351 mg of CaCl2 2H2O. Acceptance criteria: 95.0%105.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin Unit/mg of calcium chloride PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 5.57.5 in the undiluted Injection, except where the concentration is greater than 50 mg/mL, in which case this range applies to the Injection diluted with water to yield a concentration of 50 mg/mL OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, or where the label states that the Injection is not for direct injection but is to be diluted before use. The label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Endotoxin RS
ASSAY PROCEDURE Sample solution: 1 g of Calcium Chloride in 0.15 M hydrochloric acid (20:1). Transfer the solution to a 250-mL volumetric flask, and dilute with water to volume. Pipet 50 mL of the solution into a suitable container, add 100 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS until the solution is deep blue. Each mL of 0.05 M edetate disodium is equivalent to 7.351 mg of CaCl2 2H2O. Acceptance criteria: 99.0%107.0% IMPURITIES Inorganic Impurities HEAVY METALS 231: NMT 10 ppm, 2.0 g in 25 mL ALUMINUM 206 (where it is labeled as intended for use in hemodialysis): NMT 1 ppm Sample: 2.0 g of Calcium Chloride IRON, ALUMINUM, AND PHOSPHATE Sample solution: 50 mg/mL Analysis: Add 2 drops of 3 N hydrochloric acid and 1 drop of phenolphthalein TS. Then add ammonium chlorideammonium hydroxide TS, dropwise, until the solution is faintly pink, add 2 drops in excess, and heat the liquid to boiling. Acceptance criteria: No turbidity or precipitate is produced. LIMIT OF MAGNESIUM AND ALKALI SALTS Sample: 1 g Analysis: Dissolve in 50 mL of water, add 500 mg of ammonium chloride, heat the solution, and boil for 1 min. Rapidly add 40 mL of oxalic acid TS, and stir vigorously until precipitation is well established. Add immediately to the warm mixture 2 drops of methyl red TS and then 6 N ammonium hydroxide, dropwise, until the mixture is just alkaline. Cool to room temperature, transfer to a 100-mL graduated cylinder, dilute with water to 100 mL, mix, and allow to stand for 4 h or overnight. Filter, and to 50 mL of the clear filtrate in a platinum dish, add 0.5 mL of sulfuric acid, and evaporate the mixture on a steam bath to a small
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Standard solution: 5 g/mL of fluoride ion from Standard stock solution [NOTEPrepare on the day of use.] Electrode system: Use a fluoride-specific, ion-indicating electrode and a silversilver chloride reference electrode connected to a pH meter capable of measuring potentials with a minimum reproducibility of 0.2 mV (see pH 791). Standard response line: Transfer 1.0, 5.0, and 10.0 mL of the Standard solution to separate 250-mL plastic beakers, to each add 50 mL of water, 5 mL of 1 N hydrochloric acid, 10 mL of 1.0 M sodium citrate, and 10 mL of 0.2 M edetate disodium. If necessary, adjust with 1 N sodium hydroxide or 1 N hydrochloric acid to a pH of 5.5. Transfer each solution to a separate 100-mL volumetric flask, and dilute with water to volume. Transfer 50 mL of each solution to separate 250-mL plastic beakers, and measure the potential of each solution with the Electrode system. Between each reading wash the electrodes with water, and absorb any residual water by blotting the electrodes dry. Plot the logarithms of the fluoride concentrations (0.05, 0.25, and 0.50 g/mL, respectively) versus potential, in mV. Sample solution: 1.0 g of Calcium Citrate in a 100-mL beaker, add 10 mL of water and, while stirring, 10 mL of 1 N hydrochloric acid. When dissolved, boil rapidly for 1 min, transfer the solution to a 250-mL plastic beaker, and cool in ice water. Add 15 mL of 1.0 M sodium citrate and 10 mL of 0.2 M edetate disodium, and adjust with 1 N sodium hydroxide or 1 N hydrochloric acid to a pH of 5.5. Transfer this solution to a 100-mL volumetric flask, and dilute with water to volume. Analysis: Transfer 50 mL of Sample solution to a 250-mL plastic beaker, and measure the potential with the Electrode system. From the measured potential and the Standard response line determine the concentration, C, in g/mL, of fluoride ion in the Sample solution. Calculate the percentage of fluoride in the specimen taken by multiplying C by 0.01. Acceptance criteria: NMT 30 ppm Organic Impurities PROCEDURE: LIMIT OF ACID-INSOLUBLE SUBSTANCES: NMT 10 mg (0.2%) Sample solution: 5 g dissolved by heating with a mixture of 10 mL of hydrochloric acid and 50 mL of water for 30 min Analysis: Weigh the residue so obtained, filtered, washed, and dried at 105 for 2 h. SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 150 for 4 h: it loses between 10.0%13.3% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Sodium Fluoride RS
Calcium Citrate
(Comment on this Monograph)id=m11520=Calcium Citrate=Ca-Chl-Monos.pdf)
570.49 C12H10Ca3O14 4H2O 1,2,3-Propanetricarboxylic acid, 2-hydroxy-, calcium salt (2:3), tetrahydrate; Calcium citrate (3:2), tetrahydrate [5785-44-4]. DEFINITION Calcium Citrate contains four molecules of water of hydration. It contains NLT 97.5% and NMT 100.5% of Ca3(C6H5O7)2, on the previously dried basis. IDENTIFICATION A. PROCEDURE Sample solution: 40 mg/mL of Calcium citrate in 0.4 M nitric acid Analysis: To 12.5 mL of Sample solution add 1 mL of mercuric sulfate TS, heat to boiling, and add 1 mL of potassium permanganate TS. Acceptance criteria: A white precipitate is formed. B. PROCEDURE Sample: 0.5 g Analysis: Ignite completely the Sample at as low a temperature as possible, cool, and dissolve the residue in a mixture of 10 mL of water and 1 mL of glacial acetic acid. Filter, and add 10 mL of ammonium oxalate TS to the filtrate. Acceptance criteria: A voluminous white precipitate is formed, and it is soluble in hydrochloric acid. ASSAY PROCEDURE Sample solution: 350 mg of Calcium Citrate in 12 mL of 0.5 M hydrochloric acid, and dilute with water to 100 mL Analysis: While stirring, add 30 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 8.307 mg of Ca3(C6H5O7)2. Acceptance criteria: 97.5%100.5% IMPURITIES Inorganic Impurities ARSENIC, Method I 211: NMT 3 ppm Sample solution: 1 g in 5 mL of 3 N hydrochloric acid, and dilute with water to 35 mL LEAD 251: NMT 10 ppm Sample solution: 0.5 g in 20 mL of 3 N hydrochloric acid, evaporating on a steam bath to 10 mL, diluting with water to 20 mL, and cooling Analysis: Use 5 mL of Diluted Standard Lead Solution (5 g of Pb) for the test. HEAVY METALS, Method I 231: NMT 20 ppm Sample solution: 1 g in 22 mL of 11 M hydrochloric acid, add 1.5 mL of ammonium hydroxide, and dilute with water to 25 mL LIMIT OF FLUORIDE [NOTEPrepare and store all solutions in plastic containers.] Standard stock solution: 2.210 mg/mL of USP Sodium Fluoride RS
Calcium Glubionate Syrup

(Comment on this Monograph)id=m11570=Calcium Glubionate Syrup=Ca-Chl-Monos.pdf) DEFINITION Calcium Glubionate Syrup is a solution containing equimolar amounts of Calcium Gluconate and Calcium Lactobionate or with Calcium Lactobionate predominating. It contains NLT 95.0% and NMT 105.0% of the labeled amount of calcium (Ca).
USP 32
IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 2 mg/mL of calcium gluconate and 4 mg/mL of calcium lactobionate Sample solution: equivalent to 0.4 mg/mL of calcium, from Syrup Application volume: 5 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3) Spray reagent: 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add 1.0 g of ceric sulfate, swirl to dissolve, and dilute with 2 N sulfuric acid to volume Analysis Samples: Standard solution and Sample solution Proceed as directed in Thin-Layer Chromatographic Identification Test 201 and dry the plate at 100 for 20 min. Allow to cool, and spray with Spray reagent. Heat the plate at 110 for 10 min. Acceptance criteria: The two principal spots of the Sample solution correspond in color, size, and RF value to those of the Standard solution. [NOTESucrose, if present in the Sample solution, appears in the chromatogram as a spot between two principal spots.] B. IDENTIFICATION TESTSGENERAL, Calcium 191: Syrup in water (1 in 10) ASSAY PROCEDURE Sample solution: 70 mg of Ca equivalent Syrup in a suitable beaker, add 2 mL of 3 N hydrochloric acid, and dilute with water to 150 mL Analysis: While stirring with a magnetic stirrer, add 20 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 2.004 mg of Ca. Acceptance criteria: 95.0%105.0% SPECIFIC TESTS PH 791: 3.44.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at a temperature not exceeding 30, and avoid freezing.

alpha epimer of glucoheptonic acid or of a mixture of the alpha and beta epimers of glucoheptonic acid. It contains NLT 95.0% and NMT 102.0% of C14H26CaO16, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. A solution of 20 mg/mL meets the requirements of the test for Identification TestsGeneral, Calcium 191. ASSAY PROCEDURE Sample solution: 800 mg of Calcium Gluceptate in 150 mL of water containing 2 mL of 3 N hydrochloric acid Analysis: While stirring, preferably with a magnetic stirrer, add 25 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 24.52 mg of C14H26CaO16. Acceptance criteria: 95.0%102.0% of C14H26CaO16 IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion shows no more chloride than corresponds to 1.0 mL of 20.0 mM hydrochloric acid (0.07%). CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion shows no more sulfate than corresponds to 1.0 mL of 20 mN sulfuric acid (0.05%). HEAVY METALS 231: NMT 20 ppm Sample solution: 40 mg/mL, use 25 mL SPECIFIC TESTS PH 791: 6.08.0, in 100 mg/mL solution LOSS ON DRYING 731: (see Thermal Analysis 891) [NOTEThe quantity taken for the determination may be adjusted, if necessary, for instrument sensitivity. Weight loss occurring at temperatures above about 160, indicative of decomposition, is not to be interpreted as Loss on Drying.] Sample: 1025 mg Analysis: Determine the percentage of volatile substances by thermogravimetric analysis on an appropriately calibrated instrument. Heat the specimen under test at a rate of 5/min in an atmosphere of nitrogen, at a flow rate of 40 mL/min. Record the thermogram to 150. Acceptance criteria: The anhydrous form loses NMT 1.0%, the 2H2O form NMT 6.9%, and the 31/2H2O form NMT 11.4%, of its weight. REDUCING SUGARS Sample: 0.50 g Analysis: Dissolve the Sample in 10 mL of hot water, add 2 mL of 3 N hydrochloric acid, boil for about 2 min, and cool. Add 5 mL of sodium carbonate TS, allow to stand for 5 min, dilute with water to 20 mL, and filter. Add 5 mL of the clear filtrate to 2 mL of alkaline cupric tartrate TS, and boil for 1 min. Acceptance criteria: No red precipitate is formed immediately. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label to indicate whether hydrous or anhydrous; if hydrous, label to indicate also the degree of hydration. USP REFERENCE STANDARDS 11 USP Calcium Gluceptate RS
Calcium Gluceptate
(Comment on this Monograph)id=m11600=Calcium Gluceptate=Ca-Chl-Monos.pdf)
C14H26CaO16 (anhydrous) Glucoheptonic acid, calcium salt (2:1); Calcium glucoheptonate (1:2) [29039-00-7].
490.42
DEFINITION Calcium Gluceptate is anhydrous or contains varying amounts of water of hydration. It consists of the calcium salt of the
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DEFINITION Calcium Gluconate is anhydrous or contains one molecule of water of hydration. The anhydrous form contains NLT 98.0% and NMT 102.0% of C12H22CaO14, calculated on the dried basis. The monohydrate form contains NLT 99.0% and NMT 101.0% of C12H22CaO14 H2O where labeled as intended for use in preparing injectable dosage forms, and NLT 98.5% and NMT 102.0% of C12H22CaO14 H2O where labeled as not intended for use in preparing injectable dosage forms. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: It meets the requirements. Sample solution: 20 mg/mL B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 10 mg/mL of USP Potassium Gluconate RS, heating in a water bath at 60, if necessary, to dissolve Sample solution: 10 mg/mL, heating in a water bath at 60, if necessary, to dissolve Application volume: 5 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3) Spray reagent: Dissolve 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add 1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N sulfuric acid to volume, and mix. Analysis: Develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, and dry at 110 for 20 min. Allow to cool, and spray with the Spray reagent. Heat the plate at 110 for about 10 min. Acceptance criteria: The principal spot of the Sample solution corresponds in color, size, and RF value to that of the Standard solution. ASSAY PROCEDURE Sample solution: 800 mg of Calcium Gluconate in 150 mL of water containing 2 mL of 3 N hydrochloric acid Analysis: While stirring, preferably with a magnetic stirrer, add 30 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 21.52 mg of C12H22CaO14. Acceptance criteria: Anhydrous form, 98.0%102.0% of C12H22CaO14; monohydrate form, 99.0%101.0% of C12H22CaO14 H2O where labeled as intended for use in preparing injectable dosage forms, and 98.5%102.0% of C12H22CaO14 H2O where labeled as not intended for use in preparing injectable dosage forms IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion shows no more chloride than corresponds to 0.07 mL of 0.020 N hydrochloric acid (0.005%). Where it is labeled as not intended for use in the preparation of injectable dosage forms, a 1.0-g portion shows no more chloride than corresponds to 1 mL of 0.020 N hydrochloric acid (0.07%). CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion dissolved in boiling water shows no more sulfate than corresponds to 0.1 mL of 0.020 N sulfuric acid (0.005%). Where it is labeled as not intended for use in the preparation of injectable dosage forms, a 2.0-g portion dissolved in boiling water shows no more sulfate than corresponds to 1 mL of 0.020 N sulfuric acid (0.05%). ARSENIC, Method I 211 Sample solution: Dissolve 1.0 g in a mixture of 10 mL of hydrochloric acid and 20 mL of water, and dilute with
Calcium Gluceptate Injection

(Comment on this Monograph)id=m11610=Calcium Gluceptate Injection=Ca-Chl-Monos.pdf) DEFINITION Calcium Gluceptate Injection is a sterile solution of Calcium Gluceptate in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of calcium (Ca). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Transfer 5 mL of the Injection to a separator, add 10 mL of chloroform, shake, and allow the layers to separate. Draw off and discard the chloroform layer, and repeat the extraction with a second 10-mL portion of chloroform. Drain the water layer into a small beaker, evaporate to dryness, and dry in a vacuum at 60 for 16 h. B. IDENTIFICATION TESTSGENERAL, Calcium 191: A dilution of the Injection (1 in 5) meets the requirements. ASSAY PROCEDURE Sample solution: To a volume of the Injection, equivalent to 45 mg of calcium, add 2 mL of 3 N hydrochloric acid and 148 mL of water. Analysis: While stirring, preferably with a magnetic stirrer, add 15 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 2.004 mg of Ca. Acceptance criteria: 95.0%105.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: Contains NMT 0.32 USP Endotoxin Unit/mg of calcium gluceptate PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 5.67.0 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose containers, preferably of Type I or Type II glass. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, or where the label states that the Injection is not for direct injection but is to be diluted before use, the label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Calcium Gluceptate RS USP Endotoxin RS
Calcium Gluconate
(Comment on this Monograph)id=m11660=Calcium Gluconate=Ca-Chl-Monos.pdf)
C12H22CaO14 (anhydrous) D-Gluconic acid, calcium salt (2:1); Calcium D-gluconate (1:2) [299-28-5]. Monohydrate
430.37 448.39
USP 32
water to 55 mL. [NOTEThe addition of 20 mL of 7 N sulfuric acid specified under Analysis is omitted.] Acceptance criteria: Meets the requirements of the test: NMT 3 ppm HEAVY METALS, Method II 231: NMT 10 ppm [NOTEWhere Calcium Gluconate is labeled as not intended for use in the preparation of injectable dosage forms, the limit is 20 ppm.] LIMIT OF MAGNESIUM AND ALKALI METALS Sample: 1.0 g Analysis: Dissolve completely the Sample in 100 mL of boiling water. Add 10 mL of ammonium chloride TS, 1 mL of ammonium hydroxide, and 50 mL of hot (maintained at 7080) ammonium oxalate TS. Allow to stand for 4 h, dilute with water to 200 mL, and filter. Evaporate 100 mL of the filtrate to dryness, and ignite to constant weight. Acceptance criteria: The weight of the residue does not exceed 2 mg (0.4%). [NOTECalcium Gluconate labeled as not intended for use in preparing injectable dosage forms is exempt from this requirement.] LIMIT OF IRON Standard solutions: 0.2, 0.4, and 1.0 g/mL of iron, prepared as follows. Separately transfer 2.0, 4.0, and 10.0 mL of Standard Iron Solution, prepared as directed under Iron 241, to 100-mL volumetric flasks, each containing 1.37 g of calcium chloride, previously tested and shown to contain less than 5 ppm of iron, and dilute with 2 N hydrochloric acid to volume. Sample solution: Transfer 1.0 g of Calcium Gluconate to a 100-mL quartz glass flask, add 20 mL of 12 N nitric acid, and heat to boiling until fumes are evolved. Add 0.5 mL of 30% hydrogen peroxide, and heat again until fumes are evolved. Repeat this process until the volume is reduced to about 5 mL. Cool, add 1.0 mL of perchloric acid, and heat to boiling. [CautionDo not heat above 190 or evaporate to dryness because of danger of explosion.] Transfer this solution to a 25-mL volumetric flask, and dilute with 2 N hydrochloric acid to volume. Blank solution: Using 0.34 g of calcium chloride, previously tested and shown to contain less than 5 ppm of iron, instead of Calcium Gluconate, prepare as directed under Sample solution. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Lamp: Iron hollow-cathode Flame: Airacetylene Analytical wavelength: Iron emission line of 248.3 nm Analysis Samples: Standard solutions, Sample solution, and Blank solution Determine the absorbances of the Standard solutions and the Sample solution, using the Blank solution as the blank and making deuterium background corrections. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of iron, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, in g/mL, of iron in the Sample solution. Calculate the concentration of iron, in ppm, in the specimen taken: Result = F C F C = monograph correction factor, 25 = concentration of iron in the Sample solution (g/mL) [NOTECalcium Gluconate labeled as not intended for use in the preparation of injectable dosage forms is exempt from this requirement.] rU rS CS

Acceptance criteria: NMT 5 ppm LIMIT OF PHOSPHATE Standard stock solution: 0.716 mg/mL of monobasic potassium phosphate. Standard solution: Dilute 1.0 mL of Standard stock solution with water to obtain 100 mL. To 2.0 mL of this solution add 98 mL of water. Sample stock solution: To 10.0 g add 90 mL of hot water (7080), and heat to boiling, with swirling, for 10 s to obtain a clear solution. Sample solution: Dilute 1 mL of Sample stock solution with water to obtain 100 mL. Analysis: To the Standard solution and Sample solution add the 4 mL of sulfomolybdic acid TS, and mix. To both solutions add 0.1 mL of a freshly prepared mixture of 3 N hydrochloric acid and stronger acid stannous chloride TS (10:1), and mix. Acceptance criteria: After 10 min any color in the Sample solution is not more intense than that in the Standard solution (0.01%). [NOTECalcium Gluconate labeled as not intended for use in the preparation of injectable dosage forms is exempt from this requirement.] Organic Impurities PROCEDURE: LIMIT OF OXALATE [NOTEUse deionized water where water is indicated.] Mobile phase: Prepare a solution containing 0.0017 M sodium bicarbonate and 0.0018 M sodium carbonate in water. Solution A: 0.0125 M sulfuric acid Solution B: Dilute 1 mL of hydrochloric acid with water to 1200 mL. Standard solution: 1.5 g/mL of sodium oxalate in Solution B Sample solution: 20 mg/mL of Calcium Gluconate in Solution B. Sonicate if necessary. Chromatographic system (See Chromatography 621, System Suitability.) Mode: Ion chromatography Detector: Conductance Analytical column: 4-mm 25-cm; 15-m packing L12 Guard column: 4-mm 5-cm; 15-m packing L12 Anion suppressor column: Micromembrane anion suppressor column connected in series with the guard and analytical columns. The anion suppressor column is equipped with a micromembrane that separates the Mobile phase from the Solution A flowing countercurrent to the Mobile phase at a rate of about 7 mL/min. [NOTECondition the system for about 15 min with Mobile phase at a flow rate of 2 mL/min.] Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2500 theoretical plates from the analyte peak Tailing factor: NMT 1.2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of oxalate in the sample taken: Result: (rU/rS) (CS/CU) (Mr1/Mr2) F 100 = peak response for oxalate from the Sample solution = peak response for oxalate from the Standard solution = concentration of sodium oxalate in the Standard solution (g/mL)
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CU
USP 32
IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 621 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 10 mg/mL of USP Potassium Gluconate RS, heating in a water bath at 60, if necessary, to dissolve Sample solution: 10 mg/mL from Injection in water Application volume: 5 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3) Spray reagent: Dissolve 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add 1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N sulfuric acid to volume, and mix. Analysis: Develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, and dry at 110 for 20 min. Allow to cool, and spray with the Spray reagent. Heat the plate at 110 for about 10 min. Acceptance criteria: The principal spot of the Sample solution corresponds in color, size, and RF value to that of the Standard solution. B. IDENTIFICATION TESTSGENERAL, Calcium 191: It meets the requirements. Sample solution: 200 mg/mL from Injection in water ASSAY PROCEDURE Sample solution: To a volume of Injection, equivalent to 500 mg of calcium gluconate, add 2 mL of 3 N hydrochloric acid, and dilute with water to 150 mL. Analysis: While stirring, preferably with a magnetic stirrer, add 20 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 2.004 mg of Ca. Acceptance criteria: 95.0%105.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.17 USP Endotoxin Unit/mg of calcium gluconate PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 6.08.2, except that in the case where it is labeled as intended for veterinary use only and as containing boric acid, it is 2.54.5 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. LABELING: Label the Injection to indicate its content, if any, of added calcium salts, calculated as percentage of calcium in the Injection. The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, or where the label states that the Injection is not for direct injection but is to be diluted before use, the label alternatively may state the total osmolar concentration in mOsmol/mL. The labeling indicates that the Injection must be clear at the time of use, and that if crystallization has occurred, warming may redissolve the precipitate. Injection intended for veterinary use only is so labeled. If Injection contains Boric Acid, it is so labeled. USP REFERENCE STANDARDS 11 USP Endotoxin RS USP Potassium Gluconate RS
= concentration of calcium gluconate in the Sample solution (mg/mL) = molecular weight of oxalate, 88.03 Mr1 = molecular weight of sodium oxalate, 134.00 Mr2 F = unit conversion factor (g to mg), 0.001 Acceptance criteria: NMT 0.01% [NOTECalcium Gluconate labeled as not intended for use in the preparation of injectable dosage forms is exempt from this requirement.] SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 105 for 16 h: the anhydrous form loses NMT 3.0% of its weight; the monohydrate form, where labeled as intended for use in preparing injectable dosage forms, loses NMT 1.0% of its weight, and where labeled as not intended for use in preparing injectable dosage forms, loses NMT 2.0% of its weight. REDUCING SUBSTANCES Sample solution: Transfer 1.0 g to a 250-mL conical flask, dissolve in 20 mL of hot water, cool, and add 25 mL of alkaline cupric citrate TS. Cover the flask, boil gently for 5 min, accurately timed, and cool rapidly to room temperature. Analysis: Add 25 mL of 0.6 N acetic acid, 10.0 mL of 0.1 N iodine VS, and 10 mL of 3 N hydrochloric acid, and titrate with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as the endpoint is approached. Perform a blank determination, omitting the specimen, and note the difference in volumes required. Each mL of the difference in volume of 0.1 N sodium thiosulfate consumed is equivalent to 2.7 mg of reducing substances (as dextrose). Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate whether it is anhydrous or monohydrate. Where the quantity of calcium gluconate is indicated in the labeling of any solution containing Calcium Gluconate, this shall be understood to be in terms of anhydrous calcium gluconate (C12H22CaO14). Calcium Gluconate intended for use in preparing injectable dosage forms is so labeled. Calcium Gluconate not intended for use in preparing injectable dosage forms is so labeled; in addition, it may be labeled also as intended for use in preparing oral dosage forms. USP REFERENCE STANDARDS 11 USP Potassium Gluconate RS
Calcium Gluconate Injection

(Comment on this Monograph)id=m11670=Calcium Gluconate Injection=Ca-Chl-Monos.pdf) DEFINITION Calcium Gluconate Injection is a sterile solution of Calcium Gluconate in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of total Ca. The calcium is in the form of calcium gluconate, except that a small amount may be replaced with an equal amount of calcium in the form of Calcium Saccharate, or other suitable calcium salts, for the purpose of stabilization. It may contain sodium hydroxide added for adjustment of the pH. Injection intended for veterinary use only may be prepared from Calcium Gluconate solubilized with Boric Acid, or from Glyconolactone, Boric Acid, and Calcium Carbonate.
USP 32

Calcium Gluconate Tablets

(Comment on this Monograph)id=m11680=Calcium Gluconate Tablets=Ca-Chl-Monos.pdf) DEFINITION Calcium Gluconate Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of calcium gluconate (C12H22CaO14). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: It meets the requirements. Sample solution: 20 mg/mL, made from a warm, filtered solution of Tablets equivalent to 100 mg/mL of calcium gluconate in water B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 10 mg/mL of USP Potassium Gluconate RS, heating in a water bath at 60, if necessary, to dissolve Sample solution: 10 mg/mL, made from a warm, filtered solution of Tablets equivalent to 100 mg/mL of calcium gluconate Application volume: 5 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3) Spray reagent: Dissolve 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add 1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N sulfuric acid to volume, and mix. Analysis: Develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, and dry at 110 for 20 min. Allow to cool, and spray with the Spray reagent. Heat the plate at 110 for about 10 min. Acceptance criteria: The principal spot of the Sample solution corresponds in color, size, and RF value to that of the Standard solution. ASSAY PROCEDURE Sample solution: Transfer an equivalent to 500 mg of calcium gluconate, from finely powdered Tablets (NLT 20), to a suitable crucible, and ignite, gently at first, until free from carbon. Cool the crucible, add 10 mL of water, and dissolve the residue by adding sufficient 3 N hydrochloric acid, dropwise, to achieve complete solution. Transfer the solution to a suitable container, and dilute with water to 150 mL. Analysis: While stirring, preferably with a magnetic stirrer, add 20 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 21.52 mg of C12H22CaO14. Acceptance criteria: 95.0%105.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Analysis: Determine the amount of C12H22CaO14 dissolved, employing atomic absorption spectrophotometry at a wavelength of about 422.8 nm on filtered portions of the solution under test, suitably diluted with water, in comparison with a Standard solution having a known concentration of calcium in the same Medium. Tolerances: NLT 75% (Q) of the labeled amount of C12H22CaO14
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Potassium Gluconate RS
Calcium Hydroxide
(Comment on this Monograph)id=m11710=Calcium Hydroxide=Ca-Chl-Monos.pdf) Ca(OH)2 Calcium hydroxide [1305-62-0]. 74.09
DEFINITION Calcium Hydroxide contains NLT 95.0% and NMT 100.5% of Ca(OH)2. IDENTIFICATION A. PROCEDURE When mixed with three to four times its weight of water, it forms a smooth magma. The clear supernatant from the magma is alkaline to litmus. B. IDENTIFICATION TESTSGENERAL, Calcium 191: The Sample solution meets the requirements. Sample solution: Mix 1 g with 20 mL of water, and add sufficient 6 N acetic acid to dissolve the solution. ASSAY PROCEDURE Sample solution: To 1.5 g of Calcium Hydroxide in a beaker, gradually add 30 mL of 3 N hydrochloric acid. When dissolved, transfer the solution to a 500-mL volumetric flask, and rinse the beaker thoroughly, adding the rinsings to the flask. Dilute with water to volume, and mix. Pipet 50 mL of the solution into a suitable container, and add 100 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 3.705 mg of Ca(OH)2. Acceptance criteria: 95.0%100.5% IMPURITIES Inorganic Impurities HEAVY METALS 231 Sample solution: Dissolve 2.0 g in 20 mL of 3 N hydrochloric acid, and evaporate on a steam bath to dryness. Dissolve the residue in 20 mL of water, and filter. Dilute the filtrate with water to 40 mL. To 20 mL of the resulting solution, add 1 mL of 0.1 N hydrochloric acid, then add water to make 25 mL. Acceptance criteria: NMT 20 ppm LIMIT OF MAGNESIUM AND ALKALI SALTS Sample solution: 0.50 g in a mixture of 30 mL of water and 10 mL of 3 N hydrochloric acid Analysis: Heat the solution, and boil for 1 min. Rapidly add 40 mL of oxalic acid TS, and stir vigorously until precipitation is well established. Add immediately to the warm mixture 2 drops of methyl red TS and then 6 N ammonium hydroxide, dropwise, until the mixture is just alkaline. Cool to room temperature, transfer to a 100-mL graduated cylinder, dilute with water to 100 mL, mix, and allow to stand for 4 h or overnight. Filter, and to 50 mL of the clear filtrate in a platinum dish add 0.5 mL of sulfuric acid, and evaporate the mixture on a steam bath to a small volume. Carefully heat over a free flame to dryness, and continue heating to complete decomposition and
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IMPURITIES Inorganic Impurities ALKALIES AND THEIR CARBONATES: A portion of it, saturated with carbon dioxide and subsequently boiled, is neutral in reaction. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-filled, tight containers, at a temperature not exceeding 25.
volatilization of ammonium salts. Finally, ignite the residue to constant weight. Acceptance criteria: The weight of the residue does not exceed 12 mg (4.8%). LIMIT OF ACID-INSOLUBLE SUBSTANCES Sample solution: Dissolve 2.0 g in 30 mL of 4 N hydrochloric acid, and heat to boiling. Analysis: Filter the mixture, wash the residue with hot water, and ignite. Acceptance criteria: The weight of the residue is NMT 10 mg (0.5%). CARBONATE Sample: 2 g Analysis: Mix the Sample with 50 mL of water, and add an excess of 3 N hydrochloric acid to the mixture. Acceptance criteria: The addition of an excess of 3 N hydrochloric acid to the mixture does not cause more than a slight effervescence. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Calcium Lactate
(Comment on this Monograph)id=m11790=Calcium Lactate=Ca-Chl-Monos.pdf)
Calcium Hydroxide Topical Solution

(Comment on this Monograph)id=m11730=Calcium Hydroxide Topical Solution=Ca-Chl-Monos.pdf) DEFINITION Calcium Hydroxide Topical Solution is a solution containing, in each 100 mL, NLT 140 mg of calcium hydroxide [Ca(OH)2]. Prepare Calcium Hydroxide Topical Solution as follows (see Pharmaceutical CompoundingNonsterile Preparations 795):
Calcium Hydroxide Purified Water 3g 1000 mL
C6H10CaO6 xH2O (anhydrous) Propanoic acid, 2-hydroxy-, calcium salt (2:1), hydrate; Calcium lactate (1:2) hydrate [41372-22-9]. Calcium lactate (1:2) pentahydrate [5743-47-5]. Anhydrous [814-80-2].
218.22 308.30
DEFINITION Calcium Lactate contains NLT 98.0% and NMT 101.0% of C6H10CaO6, calculated on the dried basis. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: requirements Sample solution: 50 mg/mL B. INFRARED ABSORPTION 197K Meets the
Add the Calcium Hydroxide to 1000 mL of cool Purified Water, and agitate the mixture vigorously and repeatedly during 1 h. Allow the excess calcium hydroxide to settle. Dispense only the clear supernatant. [NOTEThe solubility of calcium hydroxide, which varies with the temperature at which the solution is stored, is about 170 mg/100 mL at 15, and less at a higher temperature. The official concentration is based upon a temperature of 25.] The undissolved portion of the mixture is not suitable for preparing additional quantities of Calcium Hydroxide Topical Solution. IDENTIFICATION A. It absorbs carbon dioxide from the air, and a film of calcium carbonate forms on the surface of the liquid. B. When heated, it becomes turbid, owing to the separation of calcium hydroxide. C. IDENTIFICATION TESTSGENERAL, Calcium 191: Meets the requirements ASSAY PROCEDURE Sample solution: To 100 mL of Topical Solution, add 50 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 3.705 mg of Ca(OH)2. Acceptance criteria: NLT 140 mg of Ca(OH)2 in each 100 mL of solution
ASSAY PROCEDURE Sample solution: Transfer an amount of Calcium Lactate, equivalent to 350 mg of C6H10CaO6, to a suitable container, and dissolve in a mixture of water and 3 N hydrochloric acid (150:2). Analysis: While stirring, preferably with a magnetic stirrer, add 30 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue. Continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 10.91 mg of C6H10CaO6. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities HEAVY METALS 231: NMT 20 ppm Sample solution: Dissolve 1 g in 2.5 mL of 1 N acetic acid, and dilute with water to 25 mL. LIMIT OF MAGNESIUM AND ALKALI SALTS Sample solution: Mix 1.0 g with 40 mL of water, carefully add 1 mL of hydrochloric acid, and heat the solution to boiling. Analysis: Rapidly add 40 mL of oxalic acid TS, and stir vigorously until precipitation is well established. Add immediately to the warm mixture 2 drops of methyl red TS and then 6 N ammonium hydroxide, dropwise, until the
USP 32
mixture is just alkaline. Cool to room temperature, transfer to a 100-mL graduated cylinder, dilute with water to 100 mL, mix, and allow to stand for 4 h or overnight. Filter, and to 50 mL of the clear filtrate in a platinum dish add 0.5 mL of sulfuric acid. Evaporate the mixture on a steam bath to a small volume. Carefully heat over a free flame to dryness, and continue heating to complete decomposition and volatilization of ammonium salts. Finally, ignite the residue to constant weight. Acceptance criteria: The weight of the residue does not exceed 5.0 mg (1.0%). SPECIFIC TESTS ACIDITY Sample solution: 50 mg/mL Analysis: Titrate 20 mL of Sample solution with 0.10 N sodium hydroxide, using phenolphthalein TS as the indicator. Acceptance criteria: NMT 0.50 mL is required for neutralization (0.45% as lactic acid). LOSS ON DRYING 731 Sample: 12 g Analysis: Distribute the Sample evenly in a suitable weighing dish to a depth of NMT 3 mm, and dry at 120 for 4 h. Acceptance criteria: The pentahydrate loses between 22.0% and 27.0% of its weight; the trihydrate loses between 15.0% and 20.0% of its weight; the monohydrate loses between 5.0% and 8.0% of its weight; and the dried form loses NMT 3.0% of its weight. VOLATILE FATTY ACID Sample: 500 mg Analysis: Stir the Sample with 1 mL of sulfuric acid, and warm. Acceptance criteria: The mixture does not emit an odor of volatile fatty acid. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label indicates whether it is the dried form or is hydrous; if the latter, the label indicates the degree of hydration. Where the quantity of Calcium Lactate is indicated in the labeling of any solution containing Calcium Lactate, this shall be understood to be in terms of calcium lactate pentahydrate (C6H10CaO6 5H2O). USP REFERENCE STANDARDS 11 USP Calcium Lactate RS

ASSAY PROCEDURE Sample solution: Transfer an equivalent to 350 mg of C6H10CaO6 5H2O, from finely powdered Tablets (NLT 20), to a suitable container, and add 150 mL of water and 2 mL of 3 N hydrochloric acid. Stir, using a magnetic stirrer, for 35 min. Analysis: While stirring, add 30 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 15.42 mg of C6H10CaO6 5H2O. Acceptance criteria: 94.0%106.0% PERFORMANCE TESTS DISSOLUTION 711, Procedure for a Pooled Sample Medium: Water; 500 mL Apparatus 1: 100 rpm Time: 45 min Analysis: Determine the amount of C6H10CaO6 5H2O dissolved, as directed in the Assay, making any necessary modifications. Tolerances: NLT 75% (Q) of the labeled amount of C6H10CaO6 5H2O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The quantity of Calcium Lactate stated in the labeling is in terms of calcium lactate pentahydrate.
Calcium Lactobionate
(Comment on this Monograph)id=m11827=Calcium Lactobionate=Ca-Chl-Monos.pdf)
Calcium Lactate Tablets

(Comment on this Monograph)id=m11820=Calcium Lactate Tablets=Ca-Chl-Monos.pdf) DEFINITION Calcium Lactate Tablets contain NLT 94.0% and NMT 106.0% of the labeled amount of calcium lactate (C6H10CaO6 5H2O). [NOTEAn equivalent amount of Calcium Lactate with less water of hydration may be used in place of C6H10CaO6 5H2O in preparing Calcium Lactate Tablets.] IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: Sample solution meets the requirements Sample solution: A filtered solution of Tablets, equivalent to 50 mg/mL of calcium lactate B. IDENTIFICATION TESTSGENERAL, Lactate 191: Sample solution meets the requirements Sample solution: A filtered solution of Tablets, equivalent to 50 mg/mL of calcium lactate
790.68 C24H42CaO24 2H2O D-Gluconic acid, 4-O--D-galactopyranosyl-, calcium salt (2:1), dihydrate; Lactobionic acid, calcium salt (2:1), dihydrate; Calcium lactobionate (1:2), dihydrate [110638-68-1]. DEFINITION Calcium Lactobionate contains NLT 96.0% and NMT 102.0% of C24H42CaO24 2H2O. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: The Sample solution meets the requirements. Sample solution: 20 mg/mL B. INFRARED ABSORPTION 197K C. PROCEDURE Standard solution: 10 mg/mL of USP Calcium Lactobionate RS Sample solution: 10 mg/mL Chromatographic system Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 5 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3)
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Spray reagent: Dissolve 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask. Add 1.0 g of ceric sulfate, swirl to dissolve, dilute with 2 N sulfuric acid to volume, and mix. Analysis Samples: Standard solution and Sample solution Apply the Samples to the plate. Dry the plate in a current of cool air. Place the plate in a suitable chamber lined with filter paper and previously equilibrated with the Developing solvent system. Develop until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, and dry at 100 for 20 min. Allow to cool, and spray with the Spray reagent. Heat the plate at 110 for about 10 min. Acceptance criteria: The principal spot of the Sample solution corresponds in color, size, and RF value to that of the Standard solution. ASSAY PROCEDURE Sample solution: Dissolve 0.8 g of Calcium Lactobionate in a mixture of water and 3 N hydrochloric acid (150:2). Analysis: While stirring with a magnetic stirrer, add 15 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 39.53 mg of C24H42CaO24 2H2O. Acceptance criteria: 96.0%102.0% IMPURITIES Inorganic Impurities HALIDES: A 1.2-g portion, tested as directed under Chloride and Sulfate 221, Chloride, shows no more turbidity than corresponds to 0.7 mL of 0.020 N hydrochloric acid (0.04%). SULFATE (See Chloride and Sulfate 221, Sulfate.) A 2.0-g portion dissolved in boiling water shows no more sulfate than corresponds to 1 mL of 0.020 N sulfuric acid (0.05%). HEAVY METALS 231 Sample solution: 1 g in 4 mL of 1.2 N hydrochloric acid Add water to make 25 mL, warm gently until dissolved, and cool to room temperature. Acceptance criteria: NMT 20 ppm Organic Impurities PROCEDURE: REDUCING SUBSTANCES Sample solution: Transfer 1.0 g to a 250-mL conical flask, dissolve in 20 mL of water, and add 25 mL of alkaline cupric citrate TS. Cover the flask, boil gently for 5 min, and cool rapidly to room temperature. Add 25 mL of 0.6 N acetic acid, 10.0 mL of 0.1 N iodine VS, and 10 mL of 3 N hydrochloric acid. Analysis: Titrate with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as the endpoint is approached. Perform a blank determination, omitting the specimen, and note the difference in volumes required. Each mL of the difference in volume of 0.1 N sodium thiosulfate consumed is equivalent to 2.7 mg of reducing substances (as dextrose). Acceptance criteria: NMT 1.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +26.5 Sample solution: 100 mg/mL PH 791: 5.47.4, in a solution (1 in 20) +22.0 to
Calcium Levulinate
(Comment on this Monograph)id=m11850=Calcium Levulinate=Ca-Chl-Monos.pdf)
C10H14CaO6 2H2O Pentanoic acid, 4-oxo-, calcium salt (2:1), dihydrate; Calcium levulinate (1:2) dihydrate [5743-49-7]; Anhydrous [591-64-0].
306.32 270.30
DEFINITION Calcium Levulinate contains NLT 97.5% and NMT 100.5% of C10H14CaO6, calculated on the dried basis. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements of the tests B. PROCEDURE Sample solution: 0.5 g in 5 mL of water Analysis: To Sample solution add 5 mL of 1 N sodium hydroxide, and filter. To the filtrate add 5 mL of iodine TS. Acceptance criteria: A precipitate of iodoform is produced. ASSAY PROCEDURE Sample solution: 600 mg of Calcium Levulinate in 150 mL of water containing 2 mL of 3 N hydrochloric acid Analysis: While stirring with a magnetic stirrer, add 30 mL of 0.05 M edetate disodium VS from a 50-mL buret, followed by 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to the blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 13.51 mg of C10H14CaO6. Acceptance criteria: 97.5%100.5% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion shows no more chloride than corresponds to 1.0 mL of 0.020 N hydrochloric acid (0.07%). CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion shows no more sulfate than corresponds to 1.0 mL of 0.020 N sulfuric acid (0.05%). HEAVY METALS 231: NMT 20 ppm Organic Impurities PROCEDURE: REDUCING SUGARS Sample: 0.50 g Analysis: Dissolve the Sample in 10 mL of water, add 2 mL of 3 N hydrochloric acid, boil for about 2 min, and cool. Add 5 mL of sodium carbonate TS, allow to stand for 5 min, dilute with water to 20 mL, and filter. Add 5 mL of the clear filtrate to 2 mL of alkaline cupric tartrate TS, and boil for 1 min. Acceptance criteria: No red precipitate is formed immediately. SPECIFIC TESTS MELTING RANGE OR TEMPERATURES, Class I 741: 119125 PH 791: 7.08.5, in a solution (1 in 10) LOSS ON DRYING 731: Dry it at a pressure not exceeding 5 mm of mercury at 60 for 5 h: it loses 10.5%12.0% of its weight.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Calcium Lactobionate RS
USP 32
Calcium Pantothenate
(Comment on this Monograph)id=m11910=Calcium Pantothenate=Ca-Chl-Monos.pdf)
Calcium Levulinate Injection

(Comment on this Monograph)id=m11860=Calcium Levulinate Injection=Ca-Chl-Monos.pdf) DEFINITION Calcium Levulinate Injection is a sterile solution of Calcium Levulinate in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of calcium levulinate (C10H14CaO6 2H2O). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample solution: 100 mg/mL Acceptance criteria: Meets the requirements of the tests for Calcium. B. PROCEDURE Sample solution: 0.5 g in 5 mL of water Analysis: Add 5 mL of 1 N sodium hydroxide, and filter. To the filtrate add 5 mL of iodine TS. Acceptance criteria: A precipitate of iodoform is produced. ASSAY PROCEDURE Sample solution: Transfer a volume of Injection, equivalent to 600 mg of calcium levulinate, to a 400-mL beaker, and add 2 mL of hydrochloric acid. Analysis: While stirring with a magnetic stirrer, add 30 mL of 0.05 M edetate disodium VS from a 50-mL buret, followed by 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue, and continue the titration to the blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 15.32 mg of C10H14CaO6.2H2O. Acceptance criteria: 95.0%105.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 35.70 USP Endotoxin Units/mg of calcium levulinate PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 6.08.0 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, or where the label states that the Injection is not for direct injection but is to be diluted before use, the label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Endotoxin RS
476.53 C18H32CaN2O10 -Alanine, N-(2,4-dihydroxy-3,3-dimethyl-1-oxobutyl)-, calcium salt (2:1), (R)-; Calcium D-pantothenate (1:2) [137-08-6]. DEFINITION Calcium Pantothenate is the calcium salt of the dextrorotatory isomer of pantothenic acid. It contains NLT 5.7% and NMT 6.0% of nitrogen (N), and NLT 8.2% and NMT 8.6% of calcium (Ca), both calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K: Meets the requirements B. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample solution: 50 mg/mL Acceptance criteria: Meets the requirements ASSAY CONTENT OF CALCIUM Sample solution: 800 mg of Calcium Pantothenate in 150 mL of water containing 2 mL of 3 N hydrochloric acid. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS until the solution is a distinct blue in color. Each mL of 0.05 M edetate disodium is equivalent to 2.004 mg of Ca. Acceptance criteria: 8.2%8.6% NITROGEN DETERMINATION, Method I 461: Proceed as directed, except to weigh 500 mg. Acceptance criteria: 5.7%6.0% IMPURITIES Inorganic Impurities HEAVY METALS 231: NMT 20 ppm Sample solution: 1.0 g in 25 mL of water Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Sample solution: Water Standard solution: Water. Use USP Beta Alanine RS, in place of USP Calcium Pantothenate RS, as the Standard. Eluant: Alcohol and water (65:35) Visualization: 4 Acceptance criteria: NMT 1.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +27.5 +25.0 to
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USP 32
PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Standard solution: 0.6 mg/mL of USP Calcium Pantothenate RS in Medium Mobile phase: Water and phosphoric acid (1000:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 3.0% Analysis Samples: Sample solution (filtered) and Standard solution Calculate the quantity dissolved as a percentage of the labeled amount of C18H32CaN2O10 in the portion of Tablets taken: Result = (rU/rS) (CS/CU ) 100 = peak response of calcium pantothenate from the Sample solution = peak response of calcium pantothenate from the rS Standard solution = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: NLT 75% (Q) of the labeled amount of C18H32CaN2O10 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Tablets to indicate the content of dextrorotatory calcium pantothenate. USP REFERENCE STANDARDS 11 USP Calcium Pantothenate RS
Sample solution: 50 mg/mL ALKALINITY Sample solution: Dissolve 1.0 g in 15 mL of carbon dioxide-free water in a small flask. Analysis: As soon as the solution is complete, add 1.0 mL of 0.10 N hydrochloric acid, then add 0.05 mL of phenolphthalein TS. Acceptance criteria: No pink color is produced within 5 s. LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Beta Alanine RS USP Calcium Pantothenate RS
Calcium Pantothenate Tablets

(Comment on this Monograph)id=m11940=Calcium Pantothenate Tablets=Ca-Chl-Monos.pdf) DEFINITION Calcium Pantothenate Tablets contain NLT 95.0% and NMT 115.0% of the labeled amount of the dextrorotatory isomer of calcium pantothenate (C18H32CaN2O10). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191 Sample solution: Digest a quantity of powdered Tablets, equivalent to 150 mg of calcium pantothenate, with 15 mL of 1 N sodium hydroxide, and filter. B. PROCEDURE Sample solution: 5 mL of the filtrate obtained in Identification A Analysis: Add 5 mL of 1 N hydrochloric acid and 2 drops of ferric chloride TS to the Sample solution. Acceptance criteria: A strong yellow color is produced. ASSAY CALCIUM PANTOTHENATE ASSAY 91 Sample solution: Weigh and finely powder NLT 20 Tablets. Weigh a portion of powder, equivalent to 25 mg of calcium pantothenate from powdered Tablets, and transfer, with the aid of about 50 mL of water, to a 1000-mL volumetric flask. Add 2 mL of 1 N acetic acid and 100 mL of sodium acetate solution (1 in 60), then dilute with water to volume. Make further accurate dilutions, with water, to a concentration between 0.01 g/mL and 0.04 g/mL of calcium pantothenate. Acceptance criteria: 95.0%115.0% OTHER COMPONENTS CONTENT OF CALCIUM Sample solution: Weigh and finely powder NLT 20 Tablets. Weigh a portion of powder, equivalent to 500 mg of calcium pantothenate, transfer to a suitable crucible, and ignite, gently at first, until free from carbon. Cool the crucible, add 10 mL of water, and dissolve the residue by adding sufficient 3 N hydrochloric acid, dropwise, to achieve complete solution. Transfer the solution to a suitable container, dilute with water to 150 mL, and add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue. Analysis: Titrate with 0.05 M edetate disodium VS until the solution is deep blue in color. Each mL of 0.05 M edetate disodium is equivalent to 2.004 mg of Ca. Acceptance criteria: 7.9%9.7% of the weight of C18H32CaN2O10 in the Tablets, as determined by the Assay
Racemic Calcium Pantothenate

(Comment on this Monograph)id=m11970=Racemic Calcium Pantothenate=Ca-Chl-Monos.pdf) 476.53 C18H32CaN2O10 -Alanine, N-(2,4-dihydroxy-3,3-dimethyl-1-oxobutyl)-, calcium salt (2:1), ()-; Calcium DL-pantothenate (1:2) [6381-63-1]. DEFINITION Racemic Calcium Pantothenate is a mixture of the calcium salts of the dextrorotatory and levorotatory isomers of pantothenic acid. It contains NLT 5.7% and NMT 6.0% of nitrogen (N), and NLT 8.2% and NMT 8.6% of calcium (Ca), both calculated on the dried basis. [NOTEThe physiological activity of Racemic Calcium Pantothenate is approximately one-half that of Calcium Pantothenate.]
USP 32
ASSAY CONTENT OF CALCIUM Sample: 800 mg of calcium pantothenate Analysis: Dissolve in 150 mL of water containing 2 mL of 3 N hydrochloric acid. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue. Titrate with 0.05 M edetate disodium VS until the solution is a distinct blue in color. Each mL of 0.05 M edetate disodium is equivalent to 2.004 mg of Ca. Acceptance criteria: 8.2%8.6% NITROGEN DETERMINATION, Method I 461: Proceed as directed, except to weigh accurately 500 mg. IMPURITIES Inorganic Impurities HEAVY METALS 231: NMT 20 ppm Sample solution: 1.0 g in 25 mL of water SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 0.05 to +0.05 Sample solution: 50 mg/mL, in water ALKALINITY: Dissolve 1.0 g in 15 mL of carbon dioxide-free water in a small flask. As soon as solution is complete, add 1.6 mL of 0.10 N hydrochloric acid, then add 0.05 mL of phenolphthalein TS, and mix: no pink color is produced within 5 s. LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 5.0% of its weight. OTHER REQUIREMENTS: It responds to the Identification procedures under Calcium Pantothenate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label solutions containing it in terms of the equivalent amount of dextrorotatory calcium pantothenate. USP REFERENCE STANDARDS 11 USP Calcium Pantothenate RS

DEFINITION Dibasic Calcium Phosphate Dihydrate contains two molecules of water of hydration. It contains NLT 98.0% and NMT 105.0% of dibasic calcium phosphate dihydrate (CaHPO4 2H2O). IDENTIFICATION A. PROCEDURE Sample solution: Dissolve 100 mg by warming in 10 mL of 2 N hydrochloric acid. Analysis: Add 2.5 mL of ammonia TS dropwise, with shaking, and then add 5 mL of ammonium oxalate TS. Acceptance criteria: A white precipitate is formed. B. PROCEDURE Solution A: 106 mg/mL of ammonium molybdate in water (10%) [NOTEPrepare before use.] Sample solution: 100 mg of Dibasic Calcium Phosphate Dihydrate in 5 mL of diluted nitric acid Analysis: Warm the Sample solution to 70, and add 2 mL of Solution A. Acceptance criteria: A yellow precipitate of ammonium phosphomolybdate is formed. ASSAY PROCEDURE Buffer: 53.5 g of ammonium chloride in water. Add 570 mL of ammonia water, stronger. Dilute with water to 1000 mL. Sample solution: Transfer 400 mg of Dibasic Calcium Phosphate Dihydrate to a 200-mL volumetric flask, dissolve in 12 mL of diluted hydrochloric acid with the aid of gentle heat, if necessary, and dilute with water to volume. Analysis: Combine 20.0 mL of Sample solution with 25.0 mL of 0.02 M edetate disodium VS, 50 mL of water, and 5 mL of Buffer. Add 25 mg of eriochrome black Tsodium chloride. Titrate the excess edetate disodium with 0.02 M zinc sulfate VS. Perform a blank determination in the same manner. Each mL of 0.02 M edetate disodium is equivalent to 3.442 mg of CaHPO4 2H2O. Acceptance criteria: 98.0%105.0% IMPURITIES Inorganic Impurities LOSS ON IGNITION 733: Ignite 1 g at 800 to 825 to constant weight: it loses 24.5%26.5% of its weight. CHLORIDE AND SULFATE, Chloride 221: To 0.20 g add 20 mL of water and 13 mL of diluted nitric acid, and warm gently, if necessary, until no more dissolves. Dilute to 100 mL, and filter, if necessary. To 50 mL of this solution add 1 mL of silver nitrate TS: the turbidity does not exceed that produced by 0.70 mL of 0.010 N hydrochloric acid (0.25%). CHLORIDE AND SULFATE, Sulfate 221: Dissolve 0.5 g in 5 mL of water and 5 mL of diluted hydrochloric acid, dilute with water to 100 mL, and filter, if necessary. To 20 mL of the filtrate add 1 mL of diluted hydrochloric acid, and dilute with water to 50 mL. Add 1 mL of barium chloride TS: the turbidity does not exceed that produced by 1.0 mL of 0.010 N sulfuric acid (0.5%). ARSENIC, Method I 211: NMT 3 ppm Sample solution: 1.0 g in 25 mL of 3 N hydrochloric acid, diluting with water to 55 mL: the resulting solution meets the requirements of the test, the addition of 20 mL of 7 N sulfuric acid specified under Analysis being omitted. BARIUM: Heat to boiling 0.50 g with 10 mL of water, and add 1 mL of hydrochloric acid dropwise, stirring after each addition. Allow to cool; filter, if necessary; and to the filtrate add 2 mL of potassium sulfate TS: no turbidity is produced within 10 min.
Dibasic Calcium Phosphate Dihydrate

(Comment on this Monograph)id=m12000=Dibasic Calcium Phosphate Dihydrate=Ca-Chl-Monos.pdf) Pharmacopeial Discussion Group Sign-Off Document
Attribute Definition Identification A Identification B Acid-insoluble substances Chloride Sulfate Carbonate Barium Loss on ignition Assay JP + + + + + + + + + + EP + + + + + + + + + + USP + + + + + + + + + +
Legend: + will adopt and implement; - will not stipulate. Nonharmonized attributes: Packaging and storage, Heavy metals, Limit of fluoride, Iron Specific local attributes: Identification C (EP), Lead (USP), Description (JP) CaHPO4 2H2O Phosphoric acid, calcium salt (1:1); Calcium phosphate, dihydrate (1:1) [7789-77-7].
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USP 32
HEAVY METALS, Method I 231: NMT 30 ppm Sample solution: Warm 1.3 g with 3 mL of 3 N hydrochloric acid until no more dissolves, dilute with water to 50 mL, and filter. LIMIT OF FLUORIDE: NMT 50 ppm [NOTEPrepare and store all solutions in plastic containers.] Solution A: 294 mg/mL of sodium citrate dihydrate in water Standard stock solution: 1.1052 mg/mL of USP Sodium Fluoride RS in water Standard solution: Transfer 20.0 mL of the Standard stock solution to a 100-mL volumetric flask containing 50 mL of Buffer, and dilute with water to volume. [NOTEEach mL of this solution contains 100 g of fluoride ion.] Sample: 2 g Sample solution: Transfer the Sample to a beaker containing a plastic-coated stirring bar, add 20 mL of water and 2.0 mL of hydrochloric acid, and stir until dissolved. Add 50.0 mL of Solution A and sufficient water to make 100 mL. Electrode system: Use a fluoride-specific, ion-indicating electrode and a silversilver chloride reference electrode connected to a pH meter capable of measuring potentials with a minimum reproducibility of 0.2 mV (see pH 791). Standard response line: Transfer 50.0 mL of Solution A and 2.0 mL of hydrochloric acid to a beaker, and add water to make 100 mL. Add a plastic-coated stirring bar, insert the electrodes into the solution, stir for 15 min, and read the potential, in mV. Continue stirring, and at 5-min intervals add 100, 100, 300, and 500 L of Standard solution, reading the potential 5 min after each addition. Plot the logarithms of the cumulative fluoride ion concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus potential, in mV. Analysis: Rinse and dry the electrodes, insert them into the Sample solution, stir for 5 min, and read the potential, in mV. From the measured potential and the Standard response line determine the concentration, C, in g/mL, of fluoride ion in the Sample solution. Calculate the percentage of fluoride in the sample taken by multiplying C by 0.005. Organic Impurities PROCEDURE 1: CARBONATE: Mix 1.0 g with 5 mL of carbon dioxide-free water, and immediately add 2 mL of hydrochloric acid: no effervescence occurs. PROCEDURE 2: LIMIT OF ACID-INSOLUBLE SUBSTANCES Sample solution: Dissolve 5.0 g with a mixture of 40 mL of water and 10 mL of hydrochloric acid by boiling gently for 5 min. Analysis: After cooling, collect the insoluble substance on ashless filter paper, and wash with water until the last washing does not give a reaction for chloride (no turbidity results from the addition of silver nitrate TS). Ignite to incinerate completely the residue and ashless filter paper for assay at 600 50. Acceptance criteria: The weight of the residue does not exceed 10 mg: NMT 0.2% of acid-insoluble substances is found. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. No storage requirements specified. USP REFERENCE STANDARDS 11 USP Sodium Fluoride RS
Anhydrous Dibasic Calcium Phosphate

(Comment on this Monograph)id=m12005=Anhydrous Dibasic Calcium Phosphate=Ca-Chl-Monos.pdf) Pharmacopeial Discussion Group Sign-Off Document
Attribute Definition Identification A Identification B Acid-insoluble substances Chloride Sulfate Carbonate Barium Loss on ignition Assay JP + + + + + + + + + + EP + + + + + + + + + + USP + + + + + + + + + +
Legend: + will adopt and implement; - will not stipulate. Nonharmonized attributes: Packaging and storage, Heavy metals, Limit of fluoride, Iron Specific local attributes: Identification C (EP), Lead (USP), Description (JP) CaHPO4 Phosphoric acid, calcium salt (1:1); Calcium phosphate (1:1) [7757-93-9]. 136.06
DEFINITION Anhydrous Dibasic Calcium Phosphate contains NLT 98.0% and NMT 103.0% of anhydrous dibasic calcium phosphate (CaHPO4). IDENTIFICATION A. PROCEDURE Sample solution: Dissolve 100 mg by warming in 10 mL of 2 N hydrochloric acid. Analysis: Add 2.5 mL of ammonia TS dropwise, with shaking, and then add 5 mL of ammonium oxalate TS. Acceptance criteria: A white precipitate is formed. B. PROCEDURE Solution A: 106 mg/mL of ammonium molybdate in water [NOTEPrepare before use.] Sample solution: 100 mg of sample in 5 mL of diluted nitric acid Analysis: Warm the solution to 70, and add 2 mL of Solution A. Acceptance criteria: A yellow precipitate of ammonium phosphomolybdate is formed. ASSAY PROCEDURE Solution A: To 53.5 g of ammonium chloride in a 1000-mL volumetric flask, add sufficient water to dissolve, followed by 570 mL of ammonium hydroxide, and dilute with water to volume. Sample solution: 400 mg of Anhydrous Dibasic Calcium Phosphate into a 200-mL volumetric flask, dissolve in 12 mL of 1.21M hydrochloric acid with the aid of gentle heat, if necessary, and dilute with water to volume.
USP 32
Analysis: Combine 20.0 mL of Sample solution with 80.0 mL of 0.02 M edetate disodium VS, Solution A, and water (5:1:10), and add 25 mg of eriochrome black Tsodium chloride. Titrate the excess edetate disodium with 0.02 M zinc sulfate VS. Perform a blank determination in the same manner. Each mL of 0.02 M edetate disodium is equivalent to 2.721 mg of CaHPO4. Acceptance criteria: 98.0%103.0% IMPURITIES Inorganic Impurities LOSS ON IGNITION 733: It loses 6.6%8.5% of its weight. Sample: 1 g Ignition temperature range: 800825 to constant weight CHLORIDE AND SULFATE, Chloride 221: To 0.20 g add 20 mL of water and 13 mL of diluted nitric acid, and warm gently, if necessary, until no more dissolves. Dilute to 100 mL, and filter, if necessary. To 50 mL of this solution add 1 mL of silver nitrate TS: the turbidity does not exceed that produced by 0.70 mL of 0.010 N hydrochloric acid (0.25%). CHLORIDE AND SULFATE, Sulfate 221: Dissolve 0.5 g in 5 mL of water and 5 mL of diluted hydrochloric acid, dilute with water to 100 mL, and filter, if necessary. To 20 mL of the filtrate add 1 mL of diluted hydrochloric acid, and dilute with water to 50 mL. Add 1 mL of barium chloride TS: the turbidity does not exceed that produced by 1.0 mL of 0.010 N sulfuric acid (0.5%). ARSENIC, Method I 211: NMT 3 ppm Sample solution: 1.0 g in 25 mL of 3 N hydrochloric acid, diluting with water to 55 mL: the resulting solution meets the requirements of the test, the addition of 20 mL of 7 N sulfuric acid specified under Analysis being omitted. BARIUM: Heat to boiling 0.50 g with 10 mL of water, and add 1 mL of hydrochloric acid dropwise, stirring after each addition. Allow to cool; filter, if necessary; and to the filtrate add 2 mL of potassium sulfate TS: no turbidity is produced within 10 min. HEAVY METALS, Method I 231: NMT 30 ppm Sample solution: Warm 1.3 g with 3 mL of 3 N hydrochloric acid until no more dissolves, dilute with water to 50 mL, and filter. LIMIT OF FLUORIDE: 50 ppm [NOTEPrepare and store all solutions in plastic containers.] Solution A: 294 mg/mL of sodium citrate dihydrate in water Standard stock solution: 1.1052 mg/mL of USP Sodium Fluoride RS in water Standard solution: Combine 20.0 mL of Standard stock solution and 50 mL of Solution A in a 100-mL volumetric flask, and dilute with water to volume. [NOTEEach mL of this solution contains 100 g of fluoride ion.] Electrode system: Use a fluoride-specific, ion-indicating electrode and a silversilver chloride reference electrode connected to a pH meter capable of measuring potentials with a minimum reproducibility of 0.2 mV (see pH 791). Standard response line: Transfer 50.0 mL of Solution A and 2.0 mL of hydrochloric acid to a beaker, and add water to make 100 mL. Add a plastic-coated stirring bar, insert the electrodes into the solution, stir for 15 min, and read the potential, in mV. Continue stirring, and at 5-min intervals add 100, 100, 300, and 500 L of Standard solution, reading the potential 5 min after each addition. Plot the logarithms of the cumulative fluoride ion concentrations (0.1, 0.2, 0.5, and 1.0 g/mL) versus potential, in mV. Sample: 2.0 g Anhydrous Dibasic Calcium Phosphate Sample solution: Transfer the Sample to a beaker containing a plastic-coated stirring bar, add 20 mL of water and 2.0 mL of hydrochloric acid, and stir until dissolved. Add 50.0 mL of Solution A and sufficient water to make 100 mL.

Analysis: Rinse and dry the electrodes, insert them into the Sample solution, stir for 5 min, and read the potential, in mV. From the measured potential and the Standard response line determine the concentration, C, in g/mL, of fluoride ion in the Sample solution. Calculate the percentage of fluoride in the sample taken by multiplying C by 0.005. Organic Impurities PROCEDURE 1: CARBONATE: Mix 1.0 g with 5 mL of carbon dioxide-free water, and immediately add 2 mL of hydrochloric acid: no effervescence occurs. PROCEDURE 2: LIMIT OF ACID-INSOLUBLE SUBSTANCES Sample solution: Dissolve 5.0 g in a mixture of 40 mL of water and 10 mL of hydrochloric acid by boiling gently for 5 min. Analysis: After cooling, collect the insoluble substance on ashless filter paper, and wash with water until the last washing does not give a reaction for chloride (no turbidity results from the addition of silver nitrate TS). Ignite to incinerate completely the residue and ashless filter paper for assay at 600 50. Acceptance criteria: The weight of the residue does not exceed 10 mg: NMT 0.2% of acid-insoluble substances is found. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. No storage requirements specified. USP REFERENCE STANDARDS 11 USP Sodium Fluoride RS
Dibasic Calcium Phosphate Tablets

(Comment on this Monograph)id=m12030=Dibasic Calcium Phosphate Tablets=Ca-Chl-Monos.pdf) DEFINITION Dibasic Calcium Phosphate Tablets contain NLT 92.5% and NMT 107.5% of the labeled amount of CaHPO4 2H2O. [NOTEAn equivalent amount of Dibasic Calcium Phosphate with less water of hydration may be used in place of CaHPO4 2H2O in preparing Dibasic Calcium Phosphate Tablets.] IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: Filtered portion of the Sample solution from the Assay meets the requirements. B. IDENTIFICATION TESTSGENERAL, Phosphate 191: Filtered portion of the Sample solution from the Assay, neutralized with ammonium hydroxide, meets the requirements. ASSAY PROCEDURE Sample solution: Transfer an equivalent to 1 g of dibasic calcium phosphate (dihydrate), from powdered Tablets (NLT 20), to a 100-mL volumetric flask containing 15 mL of hydrochloric acid and 10 mL of water. Heat on a steam bath, with occasional mixing, to dissolve the dibasic calcium phosphate, but not longer than 30 min. Cool, add water to volume, and mix. If the solution is not clear, filter, discarding the first 10 mL of the filtrate. Analysis: Transfer 25.0 mL of the Sample solution to a 250mL beaker equipped with a magnetic stirrer. With constant stirring, add, in the order named, 0.5 mL of triethanolamine, 300 mg of hydroxy naphthol blue, and, from a 50-mL buret, 23 mL of 0.05 M edetate disodium VS. Add 7.7 M sodium hydroxide solution until the initial red color changes to clear blue. Continue to add it dropwise until the color changes to violet, and add an additional 0.5 mL. [NOTEThe pH is 12.312.5.] Continue the titration dropwise with the 0.05 M edetate disodium VS to the
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USP 32
Exercise care to avoid any loss of particles.] Add 35 mL of 0.1 N hydrochloric acid, and shake for 30 min. Centrifuge, decanting and discarding the supernatant. Repeat the foregoing steps, using water instead of acid. Add 35 mL of a sodium bicarbonate solution (15 in 1000), and shake, venting as necessary to release any carbon dioxide liberated. Shake for 1 h, centrifuge, and decant the supernatant. Add 35 mL of sodium bicarbonate solution (15 in 1000), and shake for 1 h. Allow the tube and contents to stand overnight or until the contents have settled, and centrifuge. Withdraw the supernatant, and weigh the tube and contents. Calculate the weight of sodium bicarbonate solution absorbed. Acceptance criteria: NLT 35.0 g of the sodium bicarbonate solution is absorbed by 1.0 g of Calcium Polycarbophil, calculated on the dried basis. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
appearance of a clear blue endpoint that persists for NLT 60 s. Each mL of 0.05 M edetate disodium is equivalent to 8.604 mg of CaHPO4 2H2O. Acceptance criteria: 92.5%107.5% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 75 rpm Time: 45 min Analysis: Determine the amount of CaHPO4 2H2O dissolved, employing atomic absorption spectrophotometry at a wavelength of 422.7 nm in filtered portions of the solution under test, suitably diluted with Medium if necessary, in comparison with a standard solution having a known concentration of calcium in Medium. Tolerances: NLT 75% (Q) of the labeled amount of CaHPO4 2H2O UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The quantity of dibasic calcium phosphate stated in the labeling is in terms of CaHPO4 2H2O.
Calcium Saccharate
(Comment on this Monograph)id=m12090=Calcium Saccharate=Ca-Chl-Monos.pdf)
Calcium Polycarbophil
(Comment on this Monograph)id=m12060=Calcium Polycarbophil=Ca-Chl-Monos.pdf) Calcium polycarbophil [9003-97-8]. DEFINITION Calcium Polycarbophil is the calcium salt of polyacrylic acid cross-linked with divinyl glycol. IDENTIFICATION When tested as directed in the test for Absorbing Power, it absorbs about 35 times its original weight. OTHER COMPONENTS CONTENT OF CALCIUM Sample: 2 g Analysis: Place the Sample in a tared crucible, cover the crucible, leaving the lid slightly ajar, and place it in a muffle furnace. Heat to 600 over 2 h, increase the temperature to 1000 over 1 h, and maintain at 1000 for 1 h. Allow to cool slowly. Dissolve the residue in dilute hydrochloric acid (1 in 5), quantitatively transfer with the aid of dilute hydrochloric acid (1 in 5) to a 100-mL volumetric flask, and dilute with dilute hydrochloric acid (1 in 5) to volume. Pipet 15 mL of this solution into a 250-mL beaker, and add, while stirring with a magnetic stirrer, 100 mL of water, 20.0 mL of 0.05 M edetate disodium VS, and 300 mg of hydroxy naphthol blue. Adjust with 1 N sodium hydroxide solution to a pH of 9.09.5. Adjust with about 10 mL of 2 N sodium hydroxide to a pH of 12.4. Titrate with 0.05 M edetate disodium VS to a persistent blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 2.004 mg of Ca. Acceptance criteria: NLT 18.0% and NMT 22.0%, calculated on the dried basis SPECIFIC TESTS LOSS ON DRYING 731: Dry a sample in vacuum at 130 for 4 h: it loses NMT 10.0% of its weight. ABSORBING POWER Sample: 250 mg Analysis: Transfer the Sample to a tared 50-mL centrifuge tube fitted with a tight closure. Add 35 mL of 0.1 N hydrochloric acid to the tube, seal the tube, and shake by mechanical means for 30 min. Centrifuge at 2000 rpm for 20 min, and decant and discard the supernatant. [NOTE
D-Glucaric acid, calcium salt (1:1) tetrahydrate; Calcium D-glucarate (1:1), tetrahydrate [5793-89-5].
C6H8CaO8 4H2O
320.26
DEFINITION Calcium Saccharate is the calcium salt of D-saccharic acid. It contains NLT 98.5% and NMT 102.0% of C6H8CaO8 4H2O. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Calcium 191: Dissolve 0.2 g in 10 mL of water by the addition of 2 mL of hydrochloric acid. B. INFRARED ABSORPTION 197M ASSAY PROCEDURE Sample solution: Dissolve 600 mg of Calcium Saccharate in 150 mL of water with the aid of a sufficient volume of hydrochloric acid. While stirring, preferably with a magnetic stirrer, add about 30 mL of 0.05 M edetate disodium VS from a 50-mL buret. Add 15 mL of 1 N sodium hydroxide and 300 mg of hydroxy naphthol blue. Analysis: Continue the titration to a blue endpoint. Each mL of 0.05 M edetate disodium is equivalent to 16.01 mg of C6H8CaO8 4H2O. Acceptance criteria: 98.5%102.0% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 0.50-g portion dissolved in 10 mL of water by the addition of 2 mL of nitric acid shows no more chloride than corresponds to 0.50 mL of 0.020 N hydrochloric acid (0.07%). CHLORIDE AND SULFATE, Sulfate 221: A 0.50-g portion dissolved in 10 mL of water by the addition of 2 mL of hydrochloric acid shows no more sulfate than corresponds to 0.6 mL of 0.020 N sulfuric acid (0.12%).
USP 32
HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE: SUCROSE AND REDUCING SUGARS: Dissolve 0.5 g in 10 mL of water with the addition of 2 mL of hydrochloric acid, and boil the solution for about 2 min. Cool, add 15 mL of sodium carbonate TS, allow to stand for 5 min, and filter. Add 5 mL of the clear filtrate to about 2 mL of alkaline cupric tartrate TS, and boil for 1 min: no red precipitate is formed immediately. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +18.5 to +22.5 Sample solution: 60 mg/mL, in 4.8 N hydrochloric acid that has been allowed to stand for 1 h ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Calcium Saccharate RS
Official Monographs / Camphor 45

crystals with 75 mL of 25% alcohol: the crystals have a clean, white, glossy appearance. If not, recrystallize by dissolving the crystals in about 50 mL of alcohol. Add about 1 g of charcoal, stir, filter, and continue as directed above, beginning with Add water dropwise. Dry the crystals in a vacuum at 50 for 2 h. [NOTEIf the melting point is low, additional drying or recrystallization may be necessary.] ASSAY PROCEDURE Sample solution: Boil 40.0 mL of 0.1 N hydrochloric acid VS with 600 mg of Calcium Undecylenate for 10 min, or until the undecylenic acid layer is clear, adding water, as necessary, to maintain the original volume. Transfer the mixture, with the aid of water, to a separator. Dilute with water to about 75 mL, and extract with two 100-mL portions of solvent hexane. Wash the combined extracts with water until the last washing is neutral to litmus, and add the washings to the original water layer. Cool, and add 3 drops of methyl orange TS. Analysis: Titrate the excess hydrochloric acid with 0.1 N sodium hydroxide VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Acceptance criteria: 98.0%102.0% IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE UNDECYLENIC ACID Sample solution: 10g of Calcium Undecylenate in 250 mL of solvent hexane Mix for 2 h using a magnetic stirrer. Analysis: Filter the Sample solution, evaporate with the aid of a current of air to about 20 mL, and add 100 mL of neutralized alcohol. Add 3 drops of phenolphthalein TS, and titrate with 0.1 N sodium hydroxide VS. Acceptance criteria: NMT 0.1% SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses between 2.0% and 5.7% of its weight. PARTICLE SIZE DISTRIBUTION ESTIMATION BY ANALYTICAL SIEVING, Method I 786: Test in accordance with this procedure, except use NMT 25 g and use a single No. 100 sieve that is to be shaken for NLT 30 min or until sifting is practically complete: NLT 99.0% of it passes through a No. 100 sieve. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Calcium Undecylenate
(Comment on this Monograph)id=m12200=Calcium Undecylenate=Ca-Chl-Monos.pdf)
C22H38O4Ca 10-Undecenoic acid, calcium (2+) salt; Calcium 10-undecenoate [1322-14-1].
406.61
DEFINITION Calcium Undecylenate contains NLT 98.0% and NMT 102.0% of C22H38O4Ca (calcium undecylenate), calculated on the dried basis. IDENTIFICATION A. A filtered solution (1 in 20) in 3 N hydrochloric acid meets the requirements of the test for Identification Tests General, Calcium 191. B. MELTING RANGE OR TEMPERATURE, Class Ia 741: The crystals so obtained melt between 66 and 67.5. Sample solution: Suspend calcium undecylenate 10 g in 40 mL of water in a 250-mL separator. Cautiously and slowly add 10 mL of hydrochloric acid, while swirling. Insert the stopper, and shake. [NOTEThe separator will become quite warm, and pressure must be carefully and frequently relieved through the stopcock. If a curdy, white material remains after 5 min of shaking, add additional hydrochloric acid, 1 mL at a time, and shake until a clear oily phase is formed.] Analysis: Allow the phases to separate, drain, and discard the bottom aqueous layer. Drain and discard the middle oily layer, if present. Filter the top layer of undecylenic acid through a pledget of cotton, noting the volume obtained. To the filtrate add an equal volume of aniline. Reflux for 1 h, swirling the flask occasionally. Allow to cool, and pour 60 mL of alcohol through the condenser into the flask. Remove the flask from the condenser, add 1 g of charcoal, and stir. Filter the slurry. Add water dropwise until a few crystals form or the solution becomes slightly cloudy. [NOTEIf too much water is added, an oil will form. Add alcohol dropwise until the oil dissolves.] Allow the mixture to stand or refrigerate until crystals are formed. Collect the crystals on a filter paper inserted in a porous glass filter funnel. Wash the
Camphor
(Comment on this Monograph)id=m12230=Camphor=Ca-ChlMonos.pdf)
C10H16O Bicyclo[2.2.1]heptane-2-one, 1,7,7-trimethyl-; Camphor; 2-Bornanone [76-22-2].
152.23
DEFINITION Camphor is a ketone of Cinnamomum camphora (Linn ) Nees et e Ebermaier (Fam. Lauraceae) (Natural Camphor) or produced synthetically (Synthetic Camphor).
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Camphor / Official Monographs
USP 32
water is removed, dry the crucible and precipitate at 80 for 2 h, cool, and weigh. The weight of the precipitate so obtained, multiplied by 0.4581, represents the weight of C10H16O in the specimen taken. Acceptance criteria: 9.0 g11.0 g OTHER COMPONENTS ALCOHOL DETERMINATION, Method II 611: 80.0%87.0% of C2H5OH, the dilution to approximately 2% alcohol being made with methanol instead of with water ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
IMPURITIES Organic Impurities PROCEDURE: LIMIT OF NONVOLATILE RESIDUE: Heat 2.0 g in a tared dish on a steam bath until sublimation is complete. Dry the residue at 120 for 3 h, cool, and weigh: the weight of the residue does not exceed 1.0 mg (0.05%). Inorganic Impurities PROCEDURE: HALOGENS Sample solution: Mix 100 mg of finely divided Camphor with 200 mg of sodium peroxide in a clean, dry, hard glass test tube of about 25 mm internal diameter and 20 cm length. Suspend the tube at an angle of about 45 by means of a clamp placed at the upper end, and gently heat the tube, starting near the upper end, but not heating the clamp, and gradually bringing the heat toward the lower part of the tube until incineration is complete. Analysis: Dissolve the residue in 25 mL of warm water, acidify with nitric acid, and filter the solution into a comparison tube. Wash the test tube and the filter with two 10-mL portions of hot water, adding the washings to the filtered solution. To the filtrate, add 0.50 mL of 0.10 N silver nitrate, dilute with water to 50 mL, and mix. Acceptance criteria: The turbidity does not exceed that produced in a blank test with the same quantities of the same reagents and 0.050 mL of 0.020 N hydrochloric acid (0.035%). SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 174179 OPTICAL ROTATION, Specific Rotation 781S: +41 to +43, for natural Camphor Sample solution: 100 mg/mL, in alcohol [NOTESynthetic Camphor is optically inactive.] APPEARANCE OF SOLUTION: A 1 in 10 solution in solvent hexane is clear. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid exposure to excessive heat. LABELING: Label it to indicate whether it is of natural sources or is prepared synthetically.
Capecitabine
(Comment on this Monograph)id=m12330=Capecitabine=CaChl-Monos.pdf)
C15H22FN3O6 359.35 Carbamic acid, [1-(5-deoxy--D-ribofuranosyl)-5-fluoro-1,2dihydro-2-oxo-4-pyrimidinyl]-, pentyl ester; Pentyl 1-(5-deoxy--D-ribofuranosyl)-5-fluoro-1,2-dihydro-2oxo-4-pyrimidinecarbamate [154361-50-9]. DEFINITION Capecitabine contains NLT 98.0% and NMT 102.0% of C15H22FN3O6, calculated on the anhydrous and solvent-free basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample solution: 2 mg of sample in 300 mg of potassium bromide B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol, acetonitrile, and water (7:1:12) Solution A: 0.1% mixture of acetic acid in water Solution B: Methanol, acetonitrile, and Solution A (7:1:12) Solution C: Methanol, acetonitrile, and Solution A (16:1:3) Mobile phase: See the gradient table below.
Time (min) 0 5 20 30 31 40 Solution B (%) 100 100 49 49 100 100 Solution C (%) 0 0 51 51 0 0
Camphor Spirit
(Comment on this Monograph)id=m12240=Camphor Spirit=CaChl-Monos.pdf) DEFINITION Camphor Spirit is an alcohol solution containing, in each 100 mL, NLT 9.0 g and NMT 11.0 g of camphor (C10H16O).
Camphor Alcohol To make 100 g A sufficient quantity 1000 mL
Dissolve the camphor in about 800 mL of the alcohol, and add alcohol to make 1000 mL. Filter, if necessary. ASSAY PROCEDURE Transfer 2.0 mL of Camphor Spirit to a suitable pressure bottle containing 50 mL of freshly prepared dinitrophenylhydrazine TS. Close the pressure bottle, immerse it in a water bath, and maintain at about 75 for 16 h. Cool to room temperature, and transfer the contents to a beaker with the aid of 100 mL of 3 N sulfuric acid. Allow to stand at room temperature NLT 12 h, transfer the precipitate to a tared filter crucible, and wash with 100 mL of 3 N sulfuric acid followed by 75 mL of cold water in divided portions. Continue the suction until the excess
System suitability solution: 0.6 g/mL of USP Capecitabine RS, USP Capecitabine Related Compound A RS, USP Capecitabine Related Compound B RS, and USP Capecitabine Related Compound C RS in Diluent Standard solution: 0.6 mg/mL of USP Capecitabine RS in Diluent
USP 32
Sample solution: 0.6 mg/mL of Capecitabine in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 250 nm with a refrigerated autosampler at 5 Column: 4.6-mm 25-cm; 5-m packing L1 Column temperature: 40 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEFor the purpose of peak identification, the approximate relative retention times are given in the Impurity Table. The relative retention times are measured with respect to capecitabine.] Suitability requirements Resolution: NLT 1.0 between capecitabine related compound A and capecitabine related compound B in the System suitability solution Tailing factor: NMT 1.5 in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H22FN3O6 in the portion of Capecitabine: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Capecitabine RS in the Standard solution (mg/mL) CU = concentration of Capecitabine in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Diluent, Solution B, Solution C, System Suitability solution, Standard solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Capecitabine taken: Result = (rU/rS) (CS/CU) F 100 rU rS CS CU F = peak response for each impurity from the Sample solution = peak response for capecitabine from the Standard solution = concentration of USP Capecitabine RS in the Standard solution (mg/mL) = concentration of capecitabine in the Sample solution (mg/mL) = relative response factor for an impurity, from the Impurity Table rU rS CS
Official Monographs / Capecitabine 47

Acceptance criteria Individual impurities: See Impurity Table. Total impurities: NMT 1.5%
Impurity Table Relative Retention Time 0.18 0.19 0.36 0.95 Relative Response Factor 1.05 0.81 0.89 1.01 Acceptance Criteria, NMT (%) 0.3 0.3 0.1 0.5
Name Capecitabine related compound A Capecitabine related compound B 2,3-Di-O-acetyl-5-deoxy-5fluorocytidine 5-Deoxy-5-fluoro-N4-(2methyl-1butyloxycarbonyl)cytidine + 5-Deoxy-5-fluoro-N4-(3methyl-1butyloxycarbonyl)cytidine Capecitabine [1-[5-Deoxy-3-O-(5-deoxy-D-ribofuranosyl)--Dribofuranosyl]-5-fluoro-2oxo-1,2dihydropyrimidin-4-yl]carbamic acid pentyl ester [1-[5-Deoxy-2-O-(5-deoxy-D-ribofuranosyl)--Dribofuranosyl]-5-fluoro-2oxo-1,2dihydropyrimidin-4-yl]carbamic acid pentyl ester Capecitabine related compound C [1-[5-Deoxy-3-O-(5-deoxy-D-ribofuranosyl)--Dribofuranosyl]-5-fluoro-2oxo-1,2dihydropyrimidin-4-yl]carbamic acid pentyl ester 2,3-Di-O-acetyl-5-deoxy-5fluoro-N4(pentyloxycarbonyl)cytidine Individual unspecified impurity Total unspecified impurities
1.00 1.06
1.00 1.00
0.3
1.09
1.00
0.2
1.11 1.20
0.91 1.00
0.3 0.3
1.37
0.85
0.1
1.00
0.1 0.5
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +96.0 to +100.0 Sample solution: 10 mg/mL, on the anhydrous and solvent-free basis, in methanol, at 20 WATER DETERMINATION, Method Ic 921: NMT 0.3% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at controlled room temperature.
48
Capecitabine / Official Monographs
USP 32
Impurity Table. The relative retention times are measured with respect to capecitabine.] Suitability requirements Resolution: NLT 1.0 between capecitabine related compound A and capecitabine related compound B in the System suitability solution Tailing factor: NMT 1.5 in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H22FN3O6 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Capecitabine RS in the Standard solution (mg/mL) = nominal concentration of capecitabine in the CU Sample solution (mg/mL) Acceptance criteria: 93.0%105.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL, degassed Apparatus 2: 50 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Pass a portion of the solution under test through a 0.45-m fiberglass filter. Standard solutions: For Tablets labeled to contain 150 mg: 17 mg of USP Capecitabine RS in 100 mL of Medium For Tablets labeled to contain 500 mg: 28 mg of USP Capecitabine RS in 50 mL of Medium Analysis: Determine the amount of C15H22FN3O6 dissolved by employing UV absorption at the wavelength of maximum absorbance at 304 nm (for Tablets labeled to contain 150 mg) and at 325 nm (for Tablets labeled to contain 500 mg) on portions of the Sample solution, suitably diluted with Medium, if necessary, in comparison with the appropriate Standard solution, using a 1-mm quartz cell. Calculate the percentage of C15H22FN3O6 dissolved in each Tablet: Result = (AU/AS) CS (V/L) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of capecitabine in the Standard solution (mg/mL) V = volume of medium, 900 mL L = label claim (mg/Tablet) Tolerances: NLT 80% (Q) of the labeled amount of C15H22FN3O6 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Diluent, Acetic acid solution, Solution A, Solution B, System suitability solution, Mobile phase, Standard AU AS CS
USP REFERENCE STANDARDS 11 USP Capecitabine RS USP Capecitabine Related Compound A RS USP Capecitabine Related Compound B RS USP Capecitabine Related Compound C RS
Capecitabine Tablets
(Comment on this Monograph)id=m12335=Capecitabine Tablets=Ca-Chl-Monos.pdf) DEFINITION Capecitabine Tablets contain NLT 93.0% and NMT 105.0% of the labeled amount of capecitabine (C15H22FN3O6). IDENTIFICATION A. INFRARED ABSORPTION 197K Analytical wavelength: 15001760 cm1 Sample: Grind one Tablet to a fine powder with a mortar and pestle. Mix 1 mg of this sample with 300 mg of potassium bromide. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol, acetonitrile, and water (7:1:12) Acetic acid solution: 0.1% mixture of acetic acid in water Solution A: Methanol, acetonitrile, and Acetic acid solution (7:1:12) Solution B: Methanol, acetonitrile, and Acetic acid solution (16:1:3) System suitability solution: Includes 0.6 g/mL of USP Capecitabine RS, 0.6 g/mL of USP Capecitabine Related Compound A RS, 0.6 g/mL of USP Capecitabine Related Compound B RS, and 0.6 g/mL of USP Capecitabine Related Compound C RS in Diluent Mobile phase: See the gradient table below.
Time (min) 0 5 20 30 31 40 Solution A (%) 100 100 49 49 100 100 Solution B (%) 0 0 51 51 0 0
Standard solution: 0.6 mg/mL of USP Capecitabine RS in Diluent Sample solution: Equivalent to 0.6 mg/mL of Capecitabine, from powdered Tablets (NLT 20), in Diluent [NOTEPass through a PVDF 0.45-m membrane filter, and use the filtrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 250 nm Column: 4.6-mm 25-cm; 5-m packing L1 Column temperature: 40 Flow rate: 1 mL/min Injection size: 10 L, using a refrigerated autosampler at 5 System suitability Samples: System suitability solution and Standard solution [NOTEFor the purpose of peak identification, the approximate relative retention times are given in the
USP 32
solution, Sample solution, and Chromatographic system; Proceed as in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Tablets taken: Result = (rU/rS) (CS/CU) RRF 100 = peak response for each impurity from the Sample solution = peak response for capecitabine from the rS Standard solution = concentration of USP Capecitabine RS in the CS Standard solution (mg/mL) = concentration of capecitabine in the portion CU taken for the Sample solution as determined in the Assay (mg/mL) RRF = relative response factor for each impurity, from the Impurity Table Acceptance criteria Individual impurities: See Impurity Table. Total impurities: NMT 2.0% rU
Impurity Table Relative Retention Time 0.18 0.19 1.00 1.11 Relative Response Factor 1.05 0.81 1.00 0.91 1.00 Acceptance Criteria, NMT (%) 1.0 1.0 0.5 0.1 0.5
Official Monographs / Capreomycin 49

C25H44N14O7 652.71 Capreomycin 1A (free base); 3,6-Diamino-N-({(2S,5S,11S,15S,Z)-15-amino-2(hydroxymethyl)-11-[(R)- iminohexahydropyrimidin-4-yl] -3,6,9,12,16-pentaoxo-8-(ureidomethylene)-1,4,7,10,13pentaazacyclohexadecan-5-yl}methyl)hexanamide [37290-35-6]. Capreomycin 1B (free base); 3,6-Diamino-N-({(2S,5S,11S,15S,Z)-15-amino-11-[(R)iminohexahydropyrimidin-4-yl]-2-methyl-3,6,9,12,16pentaoxo-8-(ureidomethylene)-1,4,7,10,13pentaazacyclohexadecan-5-yl}methyl)hexanamide [33490-33-4]. DEFINITION Capreomycin Sulfate is the disulfate salt of capreomycin, a polypeptide mixture produced by the growth of Streptomyces capreolus, suitable for parenteral use. It has a potency equivalent to NLT 700 g and NMT 1050 g of capreomycin/ mg. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Sulfate 191 ASSAY PROCEDURE: Proceed as directed in AntibioticsMicrobial Assays 81, for Capreomycin Sulfate. Acceptance criteria: 7001050 g/mg IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 3.0%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid HEAVY METALS, Method II 231: NMT 30 ppm SPECIFIC TESTS CAPREOMYCIN I CONTENT Solution A: 0.5 mg/mL of ammonium bisulfate in water. Pass through a filter having a porosity of 0.5 m or less. Mobile phase: Methanol and Solution A (9:11) System suitability solution: 0.25 mg/mL of USP Capreomycin Sulfate RS in water Sample solution: 0.25 mg/mL of Capreomycin Sulfate in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 268 nm Column: 4.6-mm 15-cm; packing L10 with a 3.5% carbon loading Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution Suitability requirements Resolution: NMT 1.5 for the major peaks (capreomycin IA and capreomycin IB), System suitability solution Tailing factor: NMT 2.5 for the major peaks (capreomycin IA and capreomycin IB), System suitability solution Analysis Sample: Sample solution Calculate the percentage of capreomycin I in the portion of Capreomycin Sulfate taken: (rIA + rIB)/rT 100
Name Capecitabine related compound A Capecitabine related compound B Capecitabine Capecitabine related compound C Individual unspecified impurity Total unspecified impurities
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Capecitabine RS USP Capecitabine Related Compound A RS USP Capecitabine Related Compound B RS USP Capecitabine Related Compound C RS
Capreomycin Sulfate
(Comment on this Monograph)id=m12358=Capreomycin Sulfate=Ca-Chl-Monos.pdf)
C25H44N14O8 Capreomycin, sulfate [1405-37-4].
668.71
50
Capreomycin / Official Monographs

rIA rIB rT
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Tailing factor: NMT 2.5 for the major peaks (capreomycin IA and capreomycin IB) for the System suitability solution Analysis Sample: Sample solution Calculate the percentage of capreomycin I in the Capreomycin Sulfate: (rIA + rIB)/rT 100 = peak area for capreomycin IA = peak area for capreomycin IB = total of the peak areas for the peaks in the chromatogram Acceptance criteria: The capreomycin I content is NLT 90.0% PH 791: 4.57.5 Sample solution: 30 mg/mL LOSS ON DRYING 731: Dry 100 mg in a vacuum at a pressure not exceeding 5 mm of mercury at 100 for 4 h: it loses NMT 10.0% of its weight. BACTERIAL ENDOTOXINS TEST 85: NMT 0.35 USP Endotoxin Unit/mg of capreomycin CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. OTHER REQUIREMENTS: It also meets the requirements under Injections 1 and Identification TestsGeneral 191, Sulfate. rIA rIB rT
= peak area of capreomycin IA = peak area of capreomycin IB = total of the peak areas for the peaks in the chromatogram Acceptance criteria: NLT 90.0% PH 791: 4.57.5 Sample solution: 30 mg/mL LOSS ON DRYING 731: Dry 100 mg in a vacuum at a pressure not exceeding 5 mm of mercury at 100 for 4 h: it loses NMT 10.0% of its weight. BACTERIAL ENDOTOXINS TEST 85: Where it is intended for use in preparing injectable dosage forms: NMT 0.35 USP Endotoxin Unit/mg of capreomycin OTHER REQUIREMENTS: Where the label states that Capreomycin Sulfate is sterile, it meets the requirements under Injections 1.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Capreomycin Sulfate RS USP Endotoxin RS
Capreomycin for Injection

(Comment on this Monograph)id=m12362=Capreomycin for Injection=Ca-Chl-Monos.pdf) DEFINITION Capreomycin for Injection contains an amount of Capreomycin Sulfate equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of capreomycin. ASSAY PROCEDURE: Proceed as directed in AntibioticsMicrobial Assays 81, for Capreomycin for injection. Acceptance criteria: 90.0%115.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 3.0%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid HEAVY METALS, Method II 231: NMT 30 ppm SPECIFIC TESTS CAPREOMYCIN I CONTENT Solution A: 0.5 mg/mL of ammonium bisulfate in water. Pass through a filter having a porosity of 0.5 m or less. Mobile phase: Methanol and Solution A (9:11) System suitability solution: 0.25 mg/mL of USP Capreomycin Sulfate RS in water Sample solution: 0.25 mg/mL of Capreomycin for Injection in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 268 nm Column: 4.6-mm 15-cm; packing L10 with a 3.5% carbon loading Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution Resolution: NMT 1.5 between for the major peaks (capreomycin IA and capreomycin IB) for the System suitability solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Capreomycin Sulfate RS USP Endotoxin RS
Capsaicin
(Comment on this Monograph)id=m12365=Capsaicin=Ca-ChlMonos.pdf)
305.41 C18H27NO3 6-Nonenamide, (E)-N-[(4-Hydroxy-3-methoxy-phenyl)methyl]-8methyl; (E)-8-Methyl-N-vanillyl-6-nonenamide [404-86-4]. DEFINITION Capsaicin contains NLT 90.0% and NMT 110.0% of the labeled percentage of total capsaicinoids. The content of capsaicin (C18H27NO3) is NLT 55%, the sum of the contents of capsaicin and dihydrocapsaicin (C18H29NO3) is NLT 75%, and the content of other capsaicinoids is NMT 15%, all calculated on the dried basis. [CAUTIONHandle Capsaicin with care. Prevent inhalation of particles of it, and prevent its contact with any part of the body.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 1 mg/mL of USP Capsaicin RS in methanol
USP 32
Sample solution: 1 mg/mL of Capsaicin in methanol Application volume: 10 L Developing solvent system: Ether and methanol (19:1) Spray reagent: 0.5% solution of 2,6-dibromoquinonechlorimide in methanol Analysis Samples: Standard solution and Sample solution Analysis: Develop in the solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, and allow it to airdry. Spray the plate with Spray reagent, and allow to stand in a chamber containing ammonia fumes. Examine the chromatograms. Acceptance criteria: The blue color and the RF value of the principal spot from the Sample solution correspond to those properties of the principal spot from the Standard solution. ASSAY CONTENT OF CAPSAICIN DIHYDROCAPSAICIN, AND OTHER CAPSAICINOIDS Mobile phase: Acetonitrile and 0.015 M phosphoric acid (2:3) Standard solution A: 0.1 mg/mL of USP Capsaicin RS in methanol Standard solution B: 0.025 mg/mL of USP Dihydrocapsaicin RS in methanol Sample solution: 0.1 mg/mL of Capsaicin in methanol Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 281 nm Column: 4.6-mm 25-cm; 5-m packing L11 Column temperature: 30 [NOTEAdjust the flow rate to obtain a retention time of 20 min for the main capsaicin peak.] Injection size: 20 L System suitability Sample: Standard solution A Suitability requirements Relative standard deviation: NMT 2 Analysis Samples: Standard solution A, Standard solution B, and Sample solution Analysis: Record the chromatogram for a period of time that is twice that of the retention time of capsaicin, and measure the areas of the responses for all of the peaks. Calculate the percentage of C18H27NO3 in the portion of Capsaicin taken: (rU/rS) (CS/CU) 100 rU = peak response for capsaicin from the Sample solution rS = peak response for capsaicin from Standard solution A CS = concentration of USP Capsaicin RS in Standard solution A (mg/mL) CU = concentration of capsaicin taken to prepare the Sample solution (mg/mL) Calculate the percentage of C18H29NO3 in the portion of Capsaicin taken: (rU/rS) (CS/CU) 100 rU rS CS = peak response for dihydrocapsaicin from the Sample solution = peak response for dihydrocapsaicin from Standard solution B = concentration of USP Dihydrocapsaicin RS in Standard solution B (mg/mL) CU
Official Monographs / Capsicum 51

= concentration of capsaicin taken to prepare the Sample solution (mg/mL) Acceptance criteria: NLT 55% of capsaicin is found, and the sum of the percentage of capsaicin found and the percentage of dihydrocapsaicin found is NLT 75%. Using the chromatograms obtained from Standard solution A and the Sample solution, calculate the percentage of other capsaicinoids in the portion of Capsaicin taken: (rT/rS) (CS/CU) 100 = sum of the peak responses of the capsaicinoids, other than capsaicin and dihydrocapsaicin, from the Sample solution = peak response for capsaicin from Standard rS solution A = concentration of USP Capsaicin RS in Standard CS solution A (mg/mL) = concentration of capsaicin taken to prepare the CU Sample solution (mg/mL) Acceptance criteria: NMT 15% rT IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
NMT 1.0%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 5766; the range between beginning and end of melting does not exceed 5. LOSS ON DRYING 731: Dry it in a vacuum over phosphorus pentoxide at 40 for 5 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, and store in a cool place. LABELING: Label it to state the percentage content of total capsaicinoids. USP REFERENCE STANDARDS 11 USP Capsaicin RS USP Dihydrocapsaicin RS
Capsicum
(Comment on this Monograph)id=m12368=Capsicum=Ca-ChlMonos.pdf) DEFINITION Capsicum is the dried ripe fruit of Capsicum frutescens Linn` , e known in commerce as African Chillies, or of Capsicum annuum Linn` var. connoides Irish, known in commerce as Tabasco e Pepper, or Capsicum annuum var. longum Sendt, known in commerce as Louisiana Long Pepper, or of a hybrid between the Honka variety of Japanese Capsicum and the Old Louisiana Sport Capsicum known in commerce as Louisiana Sport Pepper (Fam. Solanaceae). SPECIFIC TESTS BOTANIC CHARACTERISTICS Unground Capsicum: This occurs as oblong-conical fruits, often curved (Louisiana Long Pepper), usually laterally compressed, 1025 mm in length and 48 mm in diameter (African Chillies), or up to 15 cm in length and 2.5 cm in diameter (Louisiana Long Pepper), or up to 5.5 cm in length and up to 13 mm in diameter (Louisiana Sport Pepper), or up to 4 cm in length and up to 9 mm in diameter (Tabasco Pepper). The fruit is two to three locular, the dissepiments being united at the base to a conical, central placenta. The pericarp is thin and membranous, its outer surface dark reddish brown to dusky yellowish orange, glabrous, shrivelled, its inner surface striate with two to three distinct
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Capsicum / Official Monographs
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. A few drops of chloroform may be added from time to time to prevent attack by insects. LABELING: Label each container to indicate which variety of Capsicum is contained therein.
longitudinal ridges representing the parietal placentae; the seeds are light brown to weak yellowish orange, suborbicular or irregular, flattened, from 2 to 4 mm in diameter, with a thickened edge and a prominent, pointed micropyle. The calyx is gamosepalous, inferior, five-toothed, and sometimes attached to a long, straight peduncle. Capsicum has a characteristic odor, and is sternutatory. Histology: The epicarp of Capsicum consists of mostly quadrangular or rectangular cells up to 80 m in length and up to 20 m deep, arranged in regular rows, with thickened and cutinized outer and radial walls, the surface of the cuticle finely striated, the radial walls somewhat wavy (African Chillies), or of polygonal, quadrangular, triangular, or irregular cells up to 76 m in length and up to 30.5 m deep (Tabasco Pepper), or up to 125 m in length and up to 38 m deep (Louisiana Long Pepper), or up to 76 m in length and up to 38 m deep (Louisiana Sport Pepper), with cuticularized outer and radial walls, the latter usually prominently beaded. The mesocarp consists of thin-walled parenchyma (African Chillies), or an outer hypodermis of tangentially elongated collenchymatous cells (Louisiana Long Pepper and Tabasco Pepper), or of from one to three rows of hypodermal cells with cuticularized walls (Louisiana Sport Pepper), a broad middle zone of thin-walled parenchyma containing yellow to red chromoplasts, oil droplets, and elaioplasts, occasionally microcrystals, and traversed by vascular bundles, and an inner zone consisting of a layer of giant cells. The endocarp consists of a layer of elongated cells, some of them very thin-walled and containing chromoplasts and others in large oval areas with thickened, beaded, lignified walls. Epidermal cells of the seed are irregular in outline and up to 342 m in length, and have very sinuous, contorted, lignified walls, the cells from the edge of the seed being much thicker walled than those from the flat surface of the seed. The embryo is curved and embedded in the endosperm, the latter consisting of smallcelled parenchyma containing fixed oil droplets and aleurone grains. Powdered Capsicum: This is a dark orange or dark reddish orange to strong yellowish brown powder. It shows numerous fragments of thin-walled parenchyma containing oil globules and orange, red, or yellow chromoplasts; and fragments of epicarp with either striated, rectangular cells arranged in parallel series (African Chillies), or with polygonal, triangular, or irregular cells, with or without beaded walls. The endocarp contains stone cells with slightly wavy, lignified walls and broad lumina. Numerous fragments of spermoderm composed of stone cells are present, showing in surface view, deeply sinuate, greatly thickened and lignified vertical walls containing numerous pore canals. Fragments of small-celled parenchyma of the endosperm containing fixed oil and aleurone grains, the latter up to 5.5 m in diameter, are also present, as well as occasional fibrovascular elements and calyx tissues. ARTICLES OF BOTANICAL ORIGIN, Acid-Insoluble Ash 561: NMT 1.25% ARTICLES OF BOTANICAL ORIGIN, Foreign Organic Matter 561: NMT 1%, other than stems and calyces, the proportion of which does not exceed 3% NONVOLATILE ETHER-SOLUBLE EXTRACTIVE: Dry a sample of Capsicum, taken as directed under Articles of Botanical Origin 561, Sampling, over phosphorus pentoxide for NLT 12 h. Extract 2.0 g of dried Capsicum with anhydrous ether for 20 h in a continuous extraction apparatus. Transfer the ether solution to a tared porcelain dish, and allow it to evaporate spontaneously. Dry the residue, still in the tared dish, over phosphorus pentoxide for 18 h, and weigh to obtain the weight of the total ether extractive. Then heat the dish gradually up to 105, until a constant weight is obtained: NLT 12% of nonvolatile ether-soluble extractive is found.
Capsicum Oleoresin
(Comment on this Monograph)id=m12370=Capsicum Oleoresin=Ca-Chl-Monos.pdf) DEFINITION Capsicum Oleoresin is an alcoholic extract of the dried ripe fruits of Capsicum annum var. minimum and small fruited varieties of C. frutescens (Solanaceae). It contains NLT 8.0% of total capsaicins [capsaicin (C18H27NO3), dihydrocapsaicin (C18H29NO3), and nordihydrocapsaicin (C17H27NO3)]. [CAUTIONCapsicum Oleoresin is a powerful irritant, and even in minute quantities produces an intense burning sensation when it comes in contact with the eyes and tender parts of the skin. Care should be taken to protect the eyes and to prevent contact of the skin with Capsicum Oleoresin.] IDENTIFICATION A. PROCEDURE Sample solution: 0.5 g of Capsicum Oleoresin in 5 mL of water and 10 mL of a mixture of water, 0.2 M potassium chloride, and 0.2 N hydrochloric acid (135:50:13), and mix. Add 5.0 mL of 0.5 M sodium nitrite and 5.0 mL of 0.02 M sodium tungstate, and mix. Analysis: Heat the Sample solution at 5560 for 15 min, allow to cool, and filter. To the filtrate add 10 mL of 1 N sodium hydroxide. Acceptance criteria: A bright yellow color is produced (presence of capsaicin). ASSAY PROCEDURE Mobile phase: Methanol and 2% acetic acid (14:11). Pass through a 0.5-m or finer porosity filter. Standard solution: 0.5 mg/mL of USP Capsaicin RS in methanol. Pass a portion of this solution through a 0.2-m porosity filter, and use the filtrate as the Standard solution. Sample solution: 10 mg/mL of Capsicum Oleoresin in methanol. Pass a portion of this solution through a 0.2-m porosity filter, and use the filtrate as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: Standard solution and Sample solution [NOTEThe relative retention times for nordihydrocapsaicin, capsaicin, and dihydrocapsaicin are about 0.9, 1.0, and 1.6, respectively.] Suitability requirements Resolution: NLT 1.2 between nordihydrocapsaicin and capsaicin in the Sample solution Tailing factor: NMT 2.0 in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of total capsaicins in the portion of Capsicum Oleoresin: Result = (rU/rS) (CS/CU) P 100 = sum of the peak areas for nordihydrocapsaicin, capsaicin, and dihydrocapsaicin from the Sample solution = peak area from the Standard solution rS = concentration of USP Capsaicin RS in the CS Standard solution (mg/mL) = concentration of Capsicum Oleoresin in the CU Sample solution (mg) P = designated purity, in percentage of USP Capsaicin RS Acceptance criteria: NLT 8.0% rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that if separation occurs, it should be warmed and mixed before use. USP REFERENCE STANDARDS 11 USP Capsaicin RS
Official Monographs / Captopril 53

System suitability solution: 10 g/mL each of USP Captopril RS, USP Captopril Disulfide RS, and 3acetylthio-2-methylpropanoic acid in methanol Standard solution: 10 g/mL of USP Captopril Disulfide RS in methanol [NOTEUse low-actinic glassware.] Sample solution: 2 mg/mL of Captopril in methanol. Use the solution promptly. [NOTEUse low-actinic glassware.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for captopril, 3acetylthio-2-methylpropanoic acid, and captopril disulfide are 0.32, 0.42, and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between captopril and 3acetylthio-2-methylpropanoic acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of captopril disulfide in the portion of Captopril taken: Result = (rU/rS) (CS/CU) 100 = peak area for captopril disulfide from the Sample solution = peak area for captopril disulfide from the rS Standard solution = concentration of USP Captopril Disulfide RS in CS the Standard solution (g/mL) = concentration of captopril in the Sample solution CU (g/mL) Acceptance criteria: NMT 1.0% of captopril disulfide. Compare the peak responses from the Sample solution, excluding those of the solvent, captopril, and captopril disulfide, with the main peak response from the Standard solution: the peak response of each impurity does not exceed 40% of the main peak response from the Standard solution (0.2%), and the sum of the impurity peak responses does not exceed the main peak response from the Standard solution (0.5%). rU SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 125 to 134 Sample solution: 10 mg/mL, in dehydrated alcohol LOSS ON DRYING 731: Dry it in a vacuum at 60 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Captopril RS USP Captopril Disulfide RS
Captopril
(Comment on this Monograph)id=m12380=Captopril=Ca-ChlMonos.pdf)
217.29 C9H15NO3S L-Proline, 1-[(2S)-3-mercapto-2-methyl-1-oxopropyl]-; 1-[(2S)-3-Mercapto-2-methylpropionyl]-L-proline [62571-86-2]. DEFINITION Captopril contains NLT 97.5% and NMT 102.0% of C9H15NO3S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K ASSAY PROCEDURE 0.1 N Potassium iodate titrant: 3.567 mg/mL of potassium iodate, previously dried at 110 to constant weight, in water Sample solution: 300 mg of Captopril in 100 mL of water in a suitable glass-stoppered flask. Add 10 mL of 3.6 N sulfuric acid, 1 g of potassium iodide, and 2 mL of starch TS. Analysis: Titrate with 0.1 N Potassium iodate titrant to a faint blue endpoint that persists for NLT 30 s. Perform a blank determination (see Titrimetry 541) and make any necessary correction. Each mL of 0.1 N Potassium iodate titrant is equivalent to 21.73 mg of C9H15NO3S. Acceptance criteria: 97.5%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% HEAVY METALS, Method II 231: NMT 30 ppm Organic Impurities PROCEDURE Mobile phase: 9 in 100 solution of tetrahydrofuran in methanol and 7.4 mM phosphoric acid (33:67)
Captopril Oral Solution

(Comment on this Monograph)id=m1599=Captopril Oral Solution=Ca-Chl-Monos.pdf) DEFINITION Captopril Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of captopril (C9H15NO3S). Prepare Captopril Oral Solution 0.75 mg/mL as follows. (See Pharmaceutical CompoundingNonsterile Preparations 795. See also Captopril Oral Suspension.)
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Captopril / Official Monographs

75 mg A sufficient quantity 100 mL
USP 32
Prepare Captopril Oral Suspension 0.75 mg/mL as follows. (See Pharmaceutical CompoundingNonsterile Preparations 795. See also Captopril Oral Solution.)
Captopril Vehicle: a mixture of Vehicle for Oral Solution (regular or sugar-free), NF, and Vehicle for Oral Suspension, NF (1:1) To make 75 mg A sufficient quantity 100 mL
Captopril powder Vehicle for Oral Solution (regular or sugar-free), NF To make
Add Captopril powder and about 10 mL of Vehicle to a mortar, and mix. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add enough Vehicle to bring to final volume, and mix well. ASSAY PROCEDURE Mobile phase: Methanol and water (11:9) containing 0.5 mL of phosphoric acid Standard solution: 7.5 g/mL of USP Captopril RS in water Sample solution: Agitate the container of Oral Solution for 30 min on a rotating mixer, remove a 5 mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the sample from the freezer, allow it to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 1.0 mL of the sample into a 100-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe retention time for Captopril is 5 min.] Suitability requirements Relative standard deviation: NMT 0.9% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H15NO3S in the volume of Oral Solution taken (mg/mL): Result = (rU/rS)(CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Captopril RS in the Standard solution (g/mL) = nominal concentration of captopril in the Sample CU solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 3.84.3 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store in a cold place. LABELING: Label to state the beyond-use date. BEYOND-USE DATE: 7 days after the day on which it was compounded USP REFERENCE STANDARDS 11 USP Captopril RS
If using Tablets, place the required number of Captopril Tablets in a suitable mortar, and comminute to a fine powder; or add Captopril powder to the mortar. Add about 10 mL of the Vehicle, and mix to a uniform paste. Add the Vehicle in small portions, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add sufficient Vehicle to bring to final volume, and mix well. ASSAY PROCEDURE Mobile phase: Methanol and water (11:9) containing 0.5 mL of phosphoric acid Standard solution: 7.5 g/mL of USP Captopril RS in water Sample solution: Agitate the container of Oral Suspension for 30 min on a rotating mixer, remove a 5mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the sample from the freezer, allow it to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 1.0 mL of the sample into a 100-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe retention time for Captopril is about 5.0 min.] Suitability requirements Relative standard deviation: NMT 0.9% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H15NO3S in the volume of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Captopril RS in the Standard solution (g/mL) = nominal concentration of captopril in the Sample CU solution (g/mL) Assay acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 3.84.3 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store in a cold place. LABELING: Label it to state that it is to be well shaken, and to state the beyond-use date. BEYOND-USE DATE: 7 days after the day on which it was compounded USP REFERENCE STANDARDS 11 USP Captopril RS rU rS CS
Captopril Oral Suspension

(Comment on this Monograph)id=m1382=Captopril Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Captopril Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of Captopril (C9H15NO3S).
USP 32
Official Monographs / Captopril 55

Captopril Tablets
(Comment on this Monograph)id=m12387=Captopril Tablets=Ca-Chl-Monos.pdf) DEFINITION Captopril Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of captopril (C9H15NO3S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 4 mg/mL, in methanol Sample solution: Equivalent to 100 mg of captopril from a portion of powdered Tablets in 25 mL of methanol in a conical flask Stir for 30 min using a magnetic stirrer, and centrifuge. Use the clear supernatant. Application volume: 50 L, as streaks Developing solvent system: Toluene, glacial acetic acid, and methanol (75:25:1) Analysis: Proceed as directed in the chapter. Locate the spots on the plate by lightly spraying with a freshly prepared mixture of 1 volume of ammonium hydroxide and 6 volumes of a solution of 0.04% 5,5-dithiobis(2nitrobenzoic acid) in methanol. ASSAY PROCEDURE [NOTEProtect solutions from exposure to air. Use within 8 h of solution.] Mobile phase: Methanol, phosphoric acid, and water (550:0.5:450) Standard solution: 1 mg/mL of USP Captopril RS and 0.05 mg/mL of USP Captopril Disulfide RS in Mobile phase Sample solution: 1 mg/mL of captopril from NLT 20 Tablets in Mobile phase to fill the flask to about half of its capacity. Sonicate for 15 min. Dilute with Mobile phase to volume, shake by mechanical means for 15 min, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 with about 15% hydrocarbon load Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for captopril and captopril disulfide are 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between captopril and captopril disulfide Relative standard deviation: NMT 2.0% Analysis: Separately inject equal volumes of the Standard solution and the Sample solution into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the percentage of C9H15NO3S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Captopril RS in the Standard solution (mg/mL) = nominal concentration of captopril in the Sample solution (mg/mL)
PERFORMANCE TESTS DISSOLUTION 711 [NOTECompletely deaerate the Medium to minimize exposure of captopril to air, and analyze the samples immediately.] Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 1: 50 rpm Time: 20 min Detector: UV 205 nm Sample solutions: Sample per Dissolution 711. Dilute with Medium to concentration similar to that of the Standard solution. Standard solution: USP Captopril RS in Medium Tolerances: NLT 80% (Q) of the labeled amount of C9H15NO3S UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF CAPTOPRIL DISULFIDE [NOTEProtect solutions from exposure to air. Use within 8 h of preparation.] Mobile phase: Proceed as directed in the Assay. System suitability solution: 1 mg/mL of USP Captopril RS and 0.05 mg/mL of USP Captopril Disulfide RS in Mobile phase Standard solution: 0.05 mg/mL of USP Captopril Disulfide RS in Mobile phase Sample solution: Equivalent to 25 mg of captopril from NLT 20 powdered Tablets in 25.0 mL of Mobile phase in a centrifuge tube Sonicate for 15 min, and centrifuge. Use the clear supernatant. Chromatographic system Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 with about 15% hydrocarbon load Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for captopril and captopril disulfide are 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between captopril and captopril disulfide in the System suitability solution Relative standard deviation: NMT 2.0% Analysis: Separately inject equal volumes of the Standard solution and the Sample solution into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the percentage of captopril disulfide in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response for captopril disulfide from the Sample solution = peak response for captopril disulfide from the Standard solution = concentration of USP Captopril Disulfide RS in the Standard solution (mg/mL) = nominal concentration of captopril in the Sample solution (mg/mL)
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Acceptance criteria: NMT 3.0% CU
USP 32
= nominal concentration of captopril and hydrochlorothiazide in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1: 50 rpm Times: 20 min for captopril; 30 min for hydrochlorothiazide Analysis: Determine the amounts of C9H15NO3S and C7H8ClN3O4S2 dissolved, using the procedure set forth in the Assay. Use filtered portions of the solution under test, making any necessary volumetric adjustments with Medium, and compare with a Standard solution having known concentrations of USP Captopril RS and USP Hydrochlorothiazide RS in the same Medium. Acceptance criteria: NLT 80% (Q) of the labeled amount of C9H15NO3S and NLT 60% (Q) of the labeled amount of C7H8ClN3O4S2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF CAPTOPRIL DISULFIDE Mobile phase: Methanol, phosphoric acid, and water (450:0.5:550) System suitability solution: 0.0075 mg/mL of USP Captopril RS and USP Hydrochlorothiazide RS, and 0.015 mg/mL of USP Captopril Disulfide RS, in Mobile phase Standard solution: 15 g/mL of USP Captopril Disulfide RS in Mobile phase Sample solution: Equivalent to 25 mg of captopril from NLT 20 powdered Tablets Add about 20 mL of Mobile phase, and sonicate for 15 min, with occasional shaking. Dilute with Mobile phase to volume, and centrifuge, made to 50 mL. Use the clear supernatant. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 30-cm; packing L11 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for captopril and captopril disulfide are 0.3 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between captopril and captopril disulfide. Both peaks are resolved from hydrochlorothiazide, System suitability solution. Relative standard deviation: NMT 3.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of captopril disulfide in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = captopril disulfide peak response from the Sample solution = captopril disulfide peak response from the Standard solution = concentration of USP Captopril Disulfide RS in the Standard solution (g/mL) = nominal concentration of captopril disulfide in the Sample solution, based on the labeled amount/Tablet (g/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Captopril RS USP Captopril Disulfide RS
Captopril and Hydrochlorothiazide Tablets

(Comment on this Monograph)id=m12398=Captopril and Hydrochlorothiazide Tablets=Ca-Chl-Monos.pdf) DEFINITION Captopril and Hydrochlorothiazide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of captopril (C9H15NO3S) and hydrochlorothiazide (C7H8ClN3O4S2). IDENTIFICATION The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, phosphoric acid, and water (250:0.5:750) System suitability solution: 0.3 mg/mL each of USP Captopril RS, USP Hydrochlorothiazide RS, and USP Benzothiadiazine Related Compound A RS in Mobile phase Standard solution: 0.3 mg/mL of USP Hydrochlorothiazide RS and about 0.3J mg/mL of USP Captopril RS, J being the ratio of the labeled amount of captopril (mg) to the labeled amount of hydrochlorothiazide/Tablet in Mobile phase (mg) Sample solution: Equivalent to 15 mg of hydrochlorothiazide from NLT 20 powdered Tablets, in Mobile phase Sonicate for 15 min with occasional shaking, and centrifuge, made to 50 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 30-cm; packing L11 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for benzothiadiazine related compound A, hydrochlorothiazide, and captopril are 0.4, 0.5, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.7 between the void volume and benzothiadiazine related compound; NLT 1.8 between benzothiadiazine related compound A and hydrochlorothiazide; and NLT 2.0 between captopril and hydrochlorothiazide Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H15NO3S and C7H8ClN3O4S2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response of the corresponding analyte from the Sample solution = peak response of the corresponding analyte from the Standard solution = concentration of the appropriate USP Reference Standard in the Standard solution (mg/mL)
USP 32
Acceptance criteria: NMT 3.0% PROCEDURE 2: LIMIT OF BENZOTHIADIAZINE RELATED COMPOUND A Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Standard solution: 10 g/mL of USP Benzothiadiazine Related Compound A RS in Mobile phase Sample solution: Use the Sample solution as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of benzothiadiazine related compound A in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response for benzothiadiazine related compound A from the Sample solution = peak response for benzothiadiazine related rs compound A from the Standard solution = concentration of USP Benzothiadiazine Related CS Compound A RS in the Standard solution (g/mL) = nominal concentration of benzothiadiazine in the CU Sample solution, based on the labeled amount/Tablet (g/mL) Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Benzothiadiazine Related Compound A RS USP Captopril RS USP Captopril Disulfide RS USP Hydrochlorothiazide RS rU
Official Monographs / Carbachol 57

perceptible when the mixture cools. Decant the supernatant, and add to the precipitate 3 mL of 3 N hydrochloric acid. Acceptance criteria: Effervescence is produced. C. IDENTIFICATION TESTSGENERAL, Calcium 191: 50 mg/mL solution D. PROCEDURE Sample solution: 100 mg/mL, in water Analysis: To 1 mL of the Sample solution, add 3 mL of gold chloride solution (1 in 10): a precipitate of yellow crystals of the aurichloride is formed. The precipitate, upon recrystallization from 5 mL of hot water, separates in glistening scale-like crystals. Dry at 105 for 1 h. Acceptance criteria: The crystals melt between 183 and 185. ASSAY PROCEDURE Sample: 400 mg of Carbachol Analysis: Dissolve the Sample in a mixture of 10 mL of glacial acetic acid and 10 mL of mercuric acetate TS. Add 2 drops of crystal violet TS, and titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 18.27 mg of C6H15ClN2O2. Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Standard solvent and Sample solvent: water (4:1) Eluant: Alcohol Visualization: 16
Methanol and
Carbachol
(Comment on this Monograph)id=m12480=Carbachol=Ca-ChlMonos.pdf)
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 200204, with some decomposition LOSS ON DRYING 731: Dry 1 g at 105 for 2 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carbachol RS
Carbachol Intraocular Solution

182.65 C6H15ClN2O2 Ethanaminium, 2-[(aminocarbonyl)oxy]-N, N,N-trimethyl-, chloride; Choline chloride, carbamate [51-83-2]. DEFINITION Carbachol contains NLT 99.0% and NMT 101.0% of C6H15ClN2O2, calculated on the dried basis. IDENTIFICATION A. PROCEDURE Sample solution: 1 mg/mL Analysis: To 5 mL of Sample solution, add 5 mL of ammonium reineckate solution (1 in 30), and shake vigorously for 1 min. Acceptance criteria: A red precipitate is formed; it is soluble in acetone. B. PROCEDURE Sample solution: 50 mg/mL in alcoholic potassium hydroxide TS Analysis: Boil 10 mL of Sample solution gently for 12 min: a white precipitate is formed, and an amine odor is (Comment on this Monograph)id=m12490=Carbachol Intraocular Solution=Ca-Chl-Monos.pdf) DEFINITION Carbachol Intraocular Solution is a sterile solution of Carbachol in an aqueous medium. It contains NLT 90.0% and NMT 115.0% of the labeled amount of carbachol (C6H15ClN2O2). It contains no preservatives or antimicrobial agents. IDENTIFICATION A. PROCEDURE Solution A: 2 mg/mL of hexanitrodiphenylamine in 0.1 N sodium hydroxide Sample solution: 100 g/mL of carbachol from Intraocular Solution in water Analysis: Transfer 5 mL of the Sample solution to a 125-mL separator. To another separator, add 5 mL of water to provide a blank. To each separator, add 1.0 mL of 1 N sodium hydroxide and 2.0 mL of Solution A. Mix, add 15 mL of methylene chloride to each separator, shake for 1 min, and allow the layers to separate.
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USP 32
Acceptance criteria: A deep amber color is produced in the methylene chloride layer from the Sample solution. ASSAY PROCEDURE Solution A: Dilute 1 volume of sodium hypochlorite solution with water to 15 volumes, allow to stand for 30 min, and mix equal volumes of the resulting solution and 1 N sodium hydroxide. [NOTEPrepare fresh daily.] Standard solution: 100 g/mL of USP Carbachol RS in water Sample solution: 100 g/mL of carbachol from a volume of Intraocular Solution in water Blank: Water Spectrometric conditions Mode: UV-Vis Cell: 1 cm Analytical wavelength: 590 nm Analysis Samples: Standard solution, Sample solution, and Blank Transfer 2.0-mL portions of the Standard solution, Sample solution, and Blank to separate 50-mL conical flasks. To each flask, add 1.0 mL of 0.1 N hydrochloric acid, and mix. Treat each as follows. Add 4.0 mL of Solution A, rinsing the inner walls of the flask with small portions of water. Mix, and allow to stand for 15 min. Add 2.0 mL of 5 mg/mL phenol solution, rinsing the walls of the flask with the solution and with additional small portions of water. Mix, and allow to stand for 5 min. Add 2.0 mL of 3.5 N hydrochloric acid, washing the sides of the flask upon addition. Rinse the flask sparingly with 0.1 N hydrochloric acid to ensure complete acidification of all contents, and mix. Add 1.0 mL of 3 mg/mL potassium iodide solution, mix, and allow to stand for 5 min. Add 3.0 mL of starch TS, mix, transfer the solutions to 50-mL volumetric flasks with the aid of several small portions of water, and dilute each solution with water to volume. Concomitantly determine the absorbances of the solutions from the Sample solution and the Standard solution against the Blank. Calculate the percentage of C6H15ClN2O2 in each mL of Intraocular Solution taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Carbachol RS in the Standard solution (g/mL) = concentration of the Sample solution (g/mL) CU Acceptance criteria: 90.0%115.0% AU AS CS SPECIFIC TESTS STERILITY TESTS 71: PH 791: 5.07.5 Meets the requirements
Carbachol Ophthalmic Solution

(Comment on this Monograph)id=m12500=Carbachol Ophthalmic Solution=Ca-Chl-Monos.pdf) DEFINITION Carbachol Ophthalmic Solution is a sterile solution of Carbachol in an isotonic, aqueous medium. It contains NLT 95.0% and NMT 105.0% of the labeled amount of carbachol (C6H15ClN2O2). It may contain suitable preservatives and antimicrobial agents. IDENTIFICATION A. PROCEDURE Sample solution: 1 mg/mL of carbachol from Ophthalmic Solution in water Analysis: Add 5 mL of ammonium reineckate solution (1 in 30), and shake vigorously for 1 min. Acceptance criteria: A red precipitate is formed; it is soluble in acetone. B. PROCEDURE Sample solution: Evaporate the equivalent of 500 mg of carbachol from a volume of Ophthalmic Solution, on a steam bath, to dryness. Analysis: To the residue, add 10 mL of alcoholic potassium hydroxide TS, and boil gently for 12 min: a white precipitate is formed, and an amine odor is perceptible when the mixture cools. Decant the supernatant, and add to the precipitate 3 mL of 3 N hydrochloric acid. Acceptance criteria: Effervescence is produced. C. PROCEDURE Sample solution: Evaporate the equivalent of 100 mg of carbachol from Ophthalmic Solution, on a steam bath, to dryness. Analysis: To the residue, add 3 mL of gold chloride solution (1 in 10): a precipitate of yellow crystals of the aurichloride is formed. The precipitate, upon recrystallization from 5 mL of hot water, separates in glistening scale-like crystals. Dry at 105 for 1 h. Acceptance criteria: The crystals melt between 183and 185. ASSAY PROCEDURE Solution A: Dilute 1 volume of sodium hypochlorite TS with water to 15 volumes, allow to stand for 30 min, and mix equal volumes of the resulting solution and 1 N sodium hydroxide. [NOTEPrepare fresh daily. ] Standard solution: 100 g/mL of USP Carbachol RS in water Sample solution: 100 g/mL of carbachol from a volume of Ophthalmic Solution in water Blank: Water Spectrometric conditions Mode: UV-Vis Cell: 1 cm Analytical wavelength: 590 nm Analysis Samples: Standard solution, Sample solution, and Blank Transfer 2.0-mL portions of the Standard solution, Sample solution, and Blank to separate 50-mL conical flasks. To each flask, add 1.0 mL of 0.1 N hydrochloric acid, and mix. Treat each as follows. Add 4.0 mL of Solution A, rinsing the inner walls of the flask with small portions of
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at controlled room temperature, and protect from freezing. LABELING: Label it to indicate that it is for single-dose intraocular use only, and that the unused portion is to be discarded. USP REFERENCE STANDARDS 11 USP Carbachol RS
USP 32
water, mix, and allow to stand for 15 min. Add 2.0 mL of 5 mg/mL phenol solution, rinsing the walls of the flask with the solution and with additional small portions of water. Mix, and allow to stand for 5 min. Add 2.0 mL of 3.5 N hydrochloric acid, washing the sides of the flask upon addition. Rinse the flask sparingly with 0.1 N hydrochloric acid to ensure complete acidification of all contents, and mix. Add 1.0 mL of 3 mg/mL potassium iodide solution, mix, and allow to stand for 5 min. Add 3.0 mL of starch TS, mix, transfer the solutions to 50-mL volumetric flasks with the aid of several small portions of water, and dilute each solution with water to volume. Concomitantly determine the absorbances of the solutions from the Sample solution and the Standard solution against the Blank. Calculate the percentage of C6H15ClN2O2 in each mL of Ophthalmic Solution taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Carbachol RS in the Standard solution (g/mL) = concentration of the Sample solution (g/mL) CU Acceptance criteria: 95.0%105.0% AU AS CS SPECIFIC TESTS STERILITY TESTS 71: PH 791: 5.07.0 Meets the requirements
Official Monographs / Carbamazepine 59

Related Compound A RS from System suitability stock solution in methanol and water (1:1) Standard stock solution: 2 mg/mL of USP Carbamazepine RS in methanol Standard solution: 0.2 mg/mL of USP Carbamazepine RS from Standard stock solution in methanol and water (1:1) Sample stock solution: 2 mg/mL of Carbamazepine in methanol Sample solution: 0.2 mg/mL of Carbamazepine from Sample stock solution in methanol and water (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; packing L10 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.70 between carbamazepine related compound A and carbamazepine from the System suitability solution Relative standard deviation: NMT 2.0% from the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H12N2O in the portion of Carbamazepine taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carbamazepine RS in the Standard solution (mg/mL) = concentration of Carbamazepine in the Sample CU solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1%, a 2.0-g sample being used CHLORIDE AND SULFATE, Chloride 221: Boil 1.0 g with 20.0 mL of water for 10 min, cool, again adjust the volume, and filter: a 10.0-mL portion of the filtrate shows no more chloride than corresponds to 0.10 mL of 0.020 N hydrochloric acid (0.014%). HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Mobile phase and System suitability solution: Proceed as directed in the Assay. Standard stock solution: 0.02 mg/mL of USP Carbamazepine RS, 0.02 mg/mL of USP Carbamazepine Related Compound A RS, and 0.02 mg/mL of USP Carbamazepine Related Compound B RS in methanol Standard solution: Dilute 5.0 mL of Standard stock solution with methanol and water (1:1) to 100.0 mL. Sample stock solution: 2 mg/mL of carbamazepine in methanol Sample solution: Transfer 25.0 mL of Sample stock solution to a 50-mL volumetric flask. Add 20 mL of water, shake, allow to cool, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carbachol RS
Carbamazepine
(Comment on this Monograph)id=m12530=Carbamazepine=Ca-Chl-Monos.pdf)
C15H12N2O Dibenz[b,f]azepine, 5-carboxamide; 5H-Dibenz[b,f]azepine-5-carboxamide [298-46-4].
236.27
DEFINITION Carbamazepine contains NLT 98.0% and NMT 102.0% of C15H12N2O, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M ASSAY PROCEDURE Mobile phase: Methanol, tetrahydrofuran, and water (12:3:85). Add 0.22 mL of formic acid to each L, mix, and then add 0.5 mL of triethylamine/L. System suitability stock solution: 0.1 mg/mL of USP Carbamazepine RS and 0.5 mg/mL of USP Carbamazepine Related Compound A RS in methanol System suitability solution: 0.01 mg/mL of USP Carbamazepine RS and 0.05 mg/mL of USP Carbamazepine
60
Carbamazepine / Official Monographs

USP Carbamazepine Related Compound A RS USP Carbamazepine Related Compound B RS
USP 32
Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; packing L10 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 1.70 between carbamazepine related compound A and carbamazepine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of carbamazepine related compound A and carbamazepine related compound B in the portion of Carbamazepine: Result = (ri/rS) (CS/CU) 100 ri = peak response for either carbamazepine related compound A or carbamazepine related compound B from the Sample solution = peak response for the corresponding peak from rS the Standard solution = concentration of carbamazepine related CS compound A or carbamazepine related compound B in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Calculate the percentage of all other impurities found in the portion of Carbamazepine: Result = (ri/rS) (CS/CU) 100 = peak response for any other impurity = peak response for carbamazepine from the Standard solution = concentration of Carbamazepine in the Standard CS solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria Individual impurities: NMT 0.2% of any individual impurity Total impurities: NMT 0.5% (including carbamazepine related compound A and carbamazepine related compound B) ri rS SPECIFIC TESTS X-RAY DIFFRACTION 941: The X-ray diffraction pattern conforms to that of USP Carbamazepine RS, similarly determined. ACIDITY Sample solution: 50 mg/mL of Carbemazepine in water [NOTESolution is prepared by mixing for 15 min and filtering.] Analysis: To a 10.0-mL aliquot of Sample solution, add 1 drop of phenolphthalein TS, and titrate with 0.01 N sodium hydroxide VS. Perform a blank determination, and make any necessary correction. NMT 1.0 mL of 0.010 N sodium hydroxide is required for each 1.0 g of Carbamazepine. ALKALINITY: To a 10.0-mL aliquot of Sample solution from Acidity, add 1 drop of methyl red TS, and titrate with 0.01 N hydrochloric acid VS. Perform a blank determination, and make any necessary correction. NMT 1.0 mL of 0.010 N hydrochloric acid is required for each 1.0 g of Carbamazepine. LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carbamazepine RS
Carbamazepine Oral Suspension

(Comment on this Monograph)id=m12550=Carbamazepine Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Carbamazepine Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of carbamazepine (C15H12N2O). IDENTIFICATION A. INFRARED ABSORPTION 197S Sample solution: Place 5 mL of Oral Suspension in a separator containing 20 mL of 0.1 N sodium hydroxide, and extract with 25 mL of chloroform. Pass the extract through anhydrous sodium sulfate supported on filter paper into a beaker. Wash the anhydrous sodium sulfate with 10 mL of chloroform, and add the washing to the extract. Evaporate the chloroform extract to dryness with the aid of a stream of nitrogen. Dissolve the residue in 10 mL of methylene chloride. ASSAY PROCEDURE Mobile phase: Methanol, tetrahydrofuran, and water (12:3:85). Add 0.22 mL of formic acid to each L, mix, and add 0.5 mL of triethylamine/L. System suitability stock solution: 0.1 mg/mL of USP Carbamazepine RS and 0.5 mg/mL of USP Carbamazepine Related Compound A RS in methanol System suitability solution: 0.01 mg/mL of USP Carbamazepine RS and 0.05 mg/mL of USP Carbamazepine Related Compound A RS from System suitability stock solution in methanol and water (1:1). Combine equal amounts of this solution and the Sample solution, accurately measured. Standard stock solution: 2 mg/mL of USP Carbamazepine RS in methanol Standard solution: 0.2 mg/mL of USP Carbamazepine RS from Standard stock solution, diluted with methanol and water (1:1) Sample stock solution: Equivalent to 200 mg of carbamazepine from a volume of freshly mixed Oral Suspension, in a 100-mL volumetric flask. Add 70 mL of methanol, shake by mechanical means for 30 min, sonicate for 2 min, dilute with methanol to volume, and mix. Allow the solution to stand for 10 min. Sample solution: Dilute 10.0 mL of the clear Sample stock solution with methanol to 100.0 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; packing L10 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.70 between carbamazepine related compound A and carbamazepine from the System suitability solution Relative standard deviation: NMT 2.0% Standard solution
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Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H12N2O in the portion of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carbamazepine RS in the Standard solution (mg/mL) = nominal concentration of carbamazepine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: The total bacterial count does not exceed 100 cfu/g, and the tests for Salmonella species and Escherichia coli are negative. PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For Oral Suspension packaged in single-unit containers: Meets the requirements DELIVERABLE VOLUME 698: For Oral Suspension packaged in multiple-unit containers: Meets the requirements. It is a performance test. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, protected from freezing and from excessive heat. USP REFERENCE STANDARDS 11 USP Carbamazepine RS USP Carbamazepine Related Compound A RS

volumetric flask, add 40 mL of methanol, and sonicate for 15 min. Allow to cool to room temperature, dilute with methanol to volume, mix, and filter, discarding the first 10 mL of the filtrate. Sample solution: 0.2 mg/mL of carbamazepine from Sample stock solution in methanol and water (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4.6-mm 25-cm; packing L10 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.70 between carbamazepine related compound A and carbamazepine from the System suitability solution Relative standard deviation: NMT 2.0% from the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H12N2O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carbamazepine RS in the Standard solution (mg/mL) = nominal concentration of carbamazepine in the CU Sample solution (mg/mL) Acceptance criteria: 92.0%108.0% PERFORMANCE TESTS DISSOLUTION 711 For products labeled as 100-mg chewable tablets Test 1: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 1. Medium: Water containing 1% sodium lauryl sulfate; 900 mL Apparatus 2: 75 rpm Time: 60 min Detector: UV 288 nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Carbamazepine RS in Medium [NOTEA volume of methanol not exceeding 1% of the final total volume of the Standard solution may be used to dissolve the carbamazepine.] Tolerances: NLT 75% (Q) of the labeled amount of C15H12N2O. Use Acceptance Table 1, with the following exceptions: at S2, no unit is less than Q 5%; at S3, no unit is less than Q 10%; and NMT 2 of the 24 units are less than Q 5%. For products labeled as 200-mg tablets Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Medium: Water containing 1% sodium lauryl sulfate; 900 mL Apparatus 2: 75 rpm Detector: UV 288 nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. rU rS CS
Carbamazepine Tablets
(Comment on this Monograph)id=m12560=Carbamazepine Tablets=Ca-Chl-Monos.pdf) DEFINITION Carbamazepine Tablets contain NLT 92.0% and NMT 108.0% of the labeled amount of carbamazepine (C15H12N2O). IDENTIFICATION A. INFRARED ABSORPTION 197M Sample solution: Boil an equivalent of 250 mg of carbamazepine, from powdered Tablets, in 15 mL of acetone for 5 min. Filter while hot, using two 5-mL portions of hot acetone to effect transfer. Evaporate the filtrate with the aid of nitrogen to 5 mL, and cool in an ice bath until crystals are formed. Filter the crystals, wash with 3 mL of cold acetone, and dry in a vacuum at 70 for 30 min. ASSAY PROCEDURE Mobile phase: Methanol, tetrahydrofuran, and water (12:3:85). Add 0.22 mL of formic acid to each L, mix, and add 0.5 mL of triethylamine/ L. System suitability stock solution: 0.1 mg/mL of USP Carbamazepine RS and 0.5 mg/mL of USP Carbamazepine Related Compound A RS in methanol System suitability solution: 0.01 mg/mL of USP Carbamazepine RS and 0.05 mg/mL of USP Carbamazepine Related Compound A RS from System suitability stock solution in methanol and water (1:1) Standard stock solution: 2 mg/mL of USP Carbamazepine RS in methanol Standard solution: 0.2 mg/mL of USP Carbamazepine RS from Standard stock solution in methanol and water (1:1) Sample stock solution: Transfer an equivalent to 100 mg of carbamazepine from NLT 20 powdered Tablets to a 50-mL
62

Standard solution: USP Carbamazepine RS in Medium [NOTEA volume of methanol not exceeding 1% of the final total volume of the Standard solution may be used to dissolve the carbamazepine.] Times: See the table below. Tolerances: See the table below.
Times (min) 15 60 Tolerances (% dissolved [Q]) 45%75% NLT 75%
USP 32
Carbamazepine Extended-Release Tablets

(Comment on this Monograph)id=m12565=Carbamazepine Extended-Release Tablets=Ca-Chl-Monos.pdf) DEFINITION Carbamazepine Extended-Release Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of carbamazepine (C15H12N2O). IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: Use the Sample solution from the test for Uniformity of Dosage Units. B. The retention time of the major PEAK of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, methylene chloride, and water (90:9:120) Internal standard solution: 600 g/mL of phenytoin in methanol Standard stock solution: 200 g/mL of USP Carbamazepine RS in methanol Standard solution: 100 g/mL of USP Carbamazepine RS from Standard stock solution in Internal standard solution System suitability solution: Internal standard solution and Standard solution (1:1) Sample stock solution: Finely powder 10 Tablets, and quantitatively transfer the powder, with the aid of alcohol, to a volumetric flask of a volume such that when the solution is diluted to volume, a final concentration estimated to be 4 mg/mL of carbamazepine is obtained. Shake by mechanical means for 60 min. Sonicate for 15 min, and dilute with methanol to volume. Allow to stand for 1015 min, then filter a portion of the supernatant, and use the clear filtrate. Sample solution A: 0.2 mg/mL from Standard stock solution in methanol Sample solution: Sample solution A and Internal standard solution (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Analytical Column: 3.9-mm 30-cm; packing L1 [NOTEWash the column with 50100 mL of methanol before and after use.] Guard column: 4.6-mm 30-mm; 7-m packing L7 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for phenytoin and carbamazepine are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.8 between phenytoin and carbamazepine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H12N2O in each Tablet taken: Result = (RU/RS) (CS/CU) 100
Use Acceptance Table 2, with the following exceptions: at 15 minat L2, no unit is more than 5% outside the stated range; at L3, no unit is more than 10% outside the stated range; and NMT 2 of the 24 units are more than 5% outside the stated range. At 60 minat L2, no unit is less than Q 5%; at L3, no unit is less than Q 10%; and NMT 2 of the 24 units are less than Q 5%. Test 3: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 3. Medium: Water containing 1% sodium lauryl sulfate; 900 mL Apparatus 2: 75 rpm Detector: UV 288 nm Sample solutions: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Standard solution: USP Carbamazepine RS in Medium [NOTEA volume of methanol not exceeding 1% of the final total volume of the Standard solution may be used to dissolve the carbamazepine.] Times: See the table below. Tolerances: See the table below.
Test 3 Table Times (min) 15 60 Tolerances (% dissolved [Q]) 60%85% NLT 75%
Use Acceptance Table 2, with the following exceptions: at 15 minat L2, no unit is more than 5% outside the stated range; at L3, no unit is more than 10% outside the stated range; and NMT 2 of the 24 units are more than 5% outside the stated range. At 60 minat L2, no unit is less than Q 5%; at L3, no unit is less than Q 10%; and NMT 2 of the 24 units are less than Q 5%. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method III 921: Dry 1.5 g of the powdered Tablets (NLT 20) at 120 for 2 h: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, preferably of glass. Dispense Carbamazepine Tablets in a container labeled Store in a dry place. Protect from moisture. LABELING: The labeling indicates the Dissolution test with which the product complies. USP REFERENCE STANDARDS 11 USP Carbamazepine RS USP Carbamazepine Related Compound A RS
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= peak response ratio of carbamazepine to the internal standard from the Sample solution = peak response ratio of carbamazepine to the RS internal standard from the Standard solution = concentration of USP Carbamazepine RS in the CS Standard solution (g/mL) = nominal concentration of carbamazepine in the CU Sample solution (g/mL) Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE 1 Standard solution: 5 g/mL of methanol and methylene chloride in dimethylformamide Sample solution: Powder 10 Tablets, and transfer to a 50mL volumetric flask. Add 30 mL of dimethylformamide, and shake by mechanical means for 60 min. Sonicate for 2 min, and dilute with dimethylformamide to volume. Centrifuge a portion at about 2500 rpm for 20 min, and use the clear supernatant. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 3-m glass; 0.2% phase G39 on support S7 Temperature Injector: 170 Detector: 300 Column: 75 for 10 min, increase 20/min to 155 and maintain at 155 for 30 min. Carrier gas: Helium Flow rate: 10 mL/min Injection volume: 2 L Analysis Samples: Standard solution and Sample solution Calculate the amount of methylane chloride and methanol, in g, in each Tablet taken: 5 CS rU/rS = concentration (g/mL) of methylene chloride or methanol in the Standard solution = peak response of the respective analyte in the rU Sample solution = peak response of the respective analyte in the rS Standard solution Acceptance criteria: NMT 23 g/Tablet of methylene chloride; NMT 100 g/Tablet of methanol PROCEDURE 2 Mobile phase: Proceed as in the Assay. System suitability solution: 60 g/mL of phenytoin and 20 g/mL of USP Carbamazepine RS in methanol Standard solution: 4 g/mL of USP Carbamazepine RS in methanol Sample solution: Use the Sample stock solution from the Assay. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Analytical column: 3.9-mm 30-cm; packing L1 [NOTEWash the column with 50100 mL of methanol before and after use.] Guard column: 4.6-mm 30-mm; 7-m packing L7 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for phenytoin and carbamazepine are about 0.8 and 1.0, respectively.] CS RU

Suitability requirements Resolution: NLT 2.8 between phenytoin and carbamazepine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of each impurity from the Sample solution = peak response for carbamazepine from the rS Standard solution = concentration of USP Carbamazepine RS in the CS Standard solution (mg/mL) = nominal concentration of carbamazepine in the CU Sample solution (mg/mL) Acceptance criteria: See Impurity Table PROCEDURE 3 Mobile phase: Methanol, acetonitrile, and water (7:3:10) System suitability solution: 12.5 g/mL of iminostilbene and 5.0 g/mL of USP Carbamazepine RS in methanol Standard solution: Use the Standard solution from Organic Impurities, Procedure 1. Sample solution: Use the Sample stock solution from the Assay. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 3.9-mm 30-cm; packing L1 [NOTEWash the column with 50100 mL of methanol before and after use.] Guard column: 4.6-mm 30-mm; 7-m packing L7 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for carbamazepine and iminostilbene are about 0.3 and 1.0, respectively.] Suitability requirements Resolution: NLT 10.0 between carbamazepine and iminostilbene Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Tablets: Result = (rU/rS) (CS/CU) 100 = peak response of each impurity from the Sample solution = peak response for carbamazepine from the rS Standard solution = concentration of USP Carbamazepine RS in the CS Standard solution (mg/mL) = nominal concentration of carbamazepine in the CU Sample solution (mg/mL) Acceptance criteria: See Impurity Table.
Impurity Table Impurity Any individual impurity Sum of all impurities (Procedure 1 + Procedure 2) Acceptance Criteria (NMT) 0.2% 0.5%
rU
rU
64
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PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL or 1800 mL for 400-mg Tablets Apparatus 1: 100 rpm Time: 3, 6, 12, and 24 h Detector: UV 284 nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Standard solution: USP Carbamazepine RS in Medium Tolerances: The percentages (Q) of the labeled amount of C15H12N2O dissolved at the times specified conform to Acceptance Table 2.
Carbamide Peroxide
(Comment on this Monograph)id=m12585=Carbamide Peroxide=Ca-Chl-Monos.pdf)
CH6N2O3 94.07 Urea compound with hydrogen peroxide (1:1) [124-43-6]. DEFINITION Carbamide Peroxide contains NLT 96.0% and NMT 102.0% of CH6N2O3. IDENTIFICATION A. Combine 1 mL of a 100 mg/mL Carbamide Peroxide solution with 1 mL of nitric acid: a white, crystalline precipitate is formed. B. IDENTIFICATION TESTSGENERAL, Peroxide 191: 100 mg/mL Carbamide Peroxide solution ASSAY PROCEDURE Sample solution: Transfer 100 mg of Carbamide Peroxide to a 500-mL iodine flask with the aid of 25 mL of water, add 5 mL of glacial acetic acid, and mix. Add 2 g of potassium iodide and 1 drop of ammonium molybdate TS, insert the stopper, and allow to stand in the dark for 10 min. Analysis: Titrate the liberated iodine in the Sample solution with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as the endpoint is approached. Each mL of 0.1 N sodium thiosulfate is equivalent to 4.704 mg of CH6N2O3. Acceptance criteria: 96.0%102.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and avoid exposure to excessive heat.
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity Standard solution: 10 g/mL of USP Carbamazepine RS in methanol Sample solution: Finely powder 1 Tablet, and quantitatively transfer the powder, with the aid of methanol, to a 100-mL volumetric flask. Add about 70 mL of methanol, and shake by mechanical means for 60 min. Sonicate for 15 min, and dilute with methanol to volume. Allow to stand for 1015 min. Dilute a portion of the clear solution with methanol to obtain a solution containing about 10 g/mL of carbamazepine. Blank: Methanol Spectrometric conditions Mode: UV Analytical wavelength: 284 nm Analysis Samples: Standard solution, Sample solution, and Blank Calculate percentage of C15H12N2O in the Tablet: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Carbamazepine RS in the Standard solution (g/mL) = nominal concentration of the Sample solution (g/mL)
Carbamide Peroxide Topical Solution

(Comment on this Monograph)id=m12590=Carbamide Peroxide Topical Solution=Ca-Chl-Monos.pdf) DEFINITION Carbamide Peroxide Topical Solution is a solution in anhydrous glycerin of Carbamide Peroxide or of carbamide peroxide prepared from hydrogen peroxide and Urea. It contains NLT 78.0% and NMT 110.0%, by weight, of the labeled amount of CH6N2O3. IDENTIFICATION A. Mix 1 mL of Topical Solution with 1 mL of nitric acid: a white, crystalline precipitate is formed.
SPECIFIC TESTS WATER DETERMINATION, Method Ia 921:
NMT 5.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Carbamazepine RS
USP 32
B. IDENTIFICATION TESTSGENERAL, Peroxide 191 ASSAY PROCEDURE Sample solution: Transfer an equivalent to 100 mg of carbamide peroxide from Topical Solution to a 500-mL iodine flask with the aid of 25 mL of water, add 5 mL of glacial acetic acid, and mix. Add 2 g of potassium iodide and 1 drop of ammonium molybdate TS, and allow to stand in the dark for 10 min. Analysis: Titrate the liberated iodine in the Sample solution with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as the endpoint is approached. Each mL of 0.1 N sodium thiosulfate is equivalent to 4.704 mg of CH6N2O3. Acceptance criteria: 78.0%110.0% SPECIFIC TESTS SPECIFIC GRAVITY 841: PH 791: 4.07.5 1.2451.272
Official Monographs / Carbenicillin 65

WATER DETERMINATION, Method I 921: NMT 6.0% BACTERIAL ENDOTOXINS TEST 85: Where it is labeled for use in preparing injectable dosage forms, NMT 0.05 USP Endotoxin Unit/mg of carbenicillin STERILITY TESTS 71: Where the label states that Carbenicillin Disodium is sterile, it meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration, 6 g being aseptically dissolved in 200 mL of Fluid A. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Carbenicillin Monosodium Monohydrate RS USP Endotoxin RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and avoid exposure to excessive heat.
Carbenicillin for Injection

(Comment on this Monograph)id=m12710=Carbenicillin for Injection=Ca-Chl-Monos.pdf) DEFINITION Carbenicillin for Injection contains an amount of Carbenicillin Disodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of carbenicillin (C17H18N2O6S). IDENTIFICATION IDENTIFICATION TESTSGENERAL, Sodium 191: requirements Meets the
Carbenicillin Disodium
(Comment on this Monograph)id=m12700=Carbenicillin Disodium=Ca-Chl-Monos.pdf)
422.36 C17H16N2Na2O6S (anhydrous) 4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6[(carboxyphenylacetyl)amino]-3,3-dimethyl-7-oxo-, disodium salt, [2S-(2,5,6)]-; N-(2-Carboxy-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]hept-6-yl)- 2-phenylmalonamic acid disodium salt [4800-94-6]. DEFINITION Carbenicillin Disodium has a potency equivalent to NLT 770 g of carbenicillin (C17H18N2O6S)/mg, calculated on the anhydrous basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Sodium 191: requirements Meets the
ASSAY PROCEDURE Buffer: Use Buffer No. 1 as described in Antibiotics Microbial Assays 81. Sample solution: Dissolve a suitable quantity of Carbenicillin Disodium in Buffer, and dilute quantitatively with Buffer to obtain a solution having a convenient concentration of carbenicillin. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81, using a volume of Sample solution diluted quantitatively with Buffer to yield a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: NLT 770 g/mL SPECIFIC TESTS PH 791: 6.58.0, in a solution containing 10 mg/mL of carbenicillin
ASSAY PROCEDURE Buffer: Use Buffer No. 1 as described in Antibiotics Microbial Assays 81. Sample solution A (single-dose): Where it is packaged for dispensing and where the package is represented as being a single-dose container, constitute Carbenicillin for Injection as directed in the labeling. Withdraw all of the withdrawable contents, and dilute quantitatively with Buffer to obtain a solution having a convenient concentration of carbenicillin. Sample solution B: Where the label states the quantity of carbenicillin in a given volume of constituted solution, constitute Carbenicillin for Injection as directed in the labeling. Dilute a measured volume of the constituted solution quantitatively with Buffer to obtain a solution having a convenient concentration of carbenicillin. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81, using a measured volume of Sample solution diluted quantitatively with Buffer to yield a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.05 USP Endotoxin Unit/mg of carbenicillin STERILITY TESTS 71: Where the label states that Carbenicillin Disodium is sterile, it meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration, 6 g being aseptically dissolved in 200 mL of Fluid A. PH 791: 6.58.0, in a solution constituted as directed in the labeling
66
Carbenicillin / Official Monographs
USP 32
Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Samples: Standard solution and Sample solution Calculate the quantity, in g, of C17H18N2O6S in each mg of Carbenicillin Indanyl Sodium taken: Result = (rU/rS) (CS/CU) P
PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections. WATER DETERMINATION, Method I 921: NMT 6.0% INJECTIONS 1: Meets the requirements for Constituted Solutions and Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Carbenicillin Monosodium Monohydrate RS USP Endotoxin RS
Carbenicillin Indanyl Sodium

(Comment on this Monograph)id=m12750=Carbenicillin Indanyl Sodium=Ca-Chl-Monos.pdf)
= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carbenicillin Indanyl Sodium RS, calculated on the anhydrous basis, in the Standard solution (g/mL) CU = concentration of Carbenicillin Indanyl Sodium in the Sample solution (g/mL) P = assigned potency of USP Carbenicillin Indanyl Sodium RS (g/mg) Acceptance criteria: 630769 g/mg SPECIFIC TESTS PH 791: 5.08.0, 100 mg/mL WATER DETERMINATION, Method I 921:
rU rS CS
C26H25N2NaO6S 516.54 4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[[3-[(2,3dihydro-1H-inden-5-yl)oxy]-1,3-dioxo-2phenylpropyl]amino]-3,3-dimethyl-7-oxo-, monosodium salt, [2S-(2,5,6)]-; 1-(5-Indanyl)(2S,5R,6R)-N-(2-carboxy-3,3-dimethyl-7-oxo-4thia-1-azabicyclo[3.2.0]hept-6-yl)-2-phenylmalonamate monosodium salt [26605-69-6]. DEFINITION Carbenicillin Indanyl Sodium has a potency equivalent to NLT 630 g and NMT 769 g of carbenicillin (C17H18N2O6S)/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sodium 191: requirements
NMT 2.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. For periods up to 18 months, store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Carbenicillin Indanyl Sodium RS
Carbenicillin Indanyl Sodium Tablets

(Comment on this Monograph)id=m12780=Carbenicillin Indanyl Sodium Tablets=Ca-Chl-Monos.pdf) DEFINITION Carbenicillin Indanyl Sodium Tablets contain the equivalent of NLT 90.0% and NMT 120.0%of the labeled amount of carbenicillin (C17H18N2O6S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Diluent: Acetone, ethyl acetate, pyridine, glacial acetic acid, and water (200:100:25:1.5:75) Standard solution: 1 mg/mL of USP Carbenicillin Indanyl Sodium RS in Diluent Sample stock solution: Equivalent to 10 mg/mL of carbenicillin from powdered Tablets in Diluent. Shake the mixture for 5 min. Sample solution: 1 mg/mL of carbenicillin, from Sample stock solution, in Diluent Application volume: 10 L Developing solvent system: Acetone, ethyl acetate, pyridine, glacial acetic acid, and water (400:300:25:2:75)
Meets the
ASSAY PROCEDURE Solution A: 0.302 mg/mL of tetrabutylammonium phosphate and 13.4 mg/mL of dibasic sodium phosphate in water, adjusted with phosphoric acid to a pH of 3.8 before final dilution Mobile phase: Acetonitrile and Solution A (21:29). Allow to stand for 1 h, and if necessary, readjust with phosphoric acid to a pH of 3.8 Diluent: Acetonitrile and 5 mM monobasic potassium phosphate (17:3) Standard solution: 250 g/mL of USP Carbenicillin Indanyl Sodium RS in Diluent [NOTEThe Standard solution contains the equivalent of 222 g/mL of carbenicillin.] Sample solution: 250 g/mL of Carbenicillin Indanyl Sodium in Diluent. Sonication may be necessary to dissolve. Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Spray reagent: Methanol, 10 mg/mL of ferric chloride in 0.1 N hydrochloric acid, and 10 mg/mL of potassium ferricyanide solution (3:4:4) Analysis Samples: Standard solution and Sample solution Develop until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and heat the plate at 80 for 30 min. Allow the plate to cool, and expose it to iodine vapors in a closed chamber for 30 s. Spray the plate with Spray reagent. Acceptance criteria: The principal spots from the Standard solution and the Sample solution are blue on a yellow-green background, and the RF value of the principal spot from the Sample solution corresponds to that from the Standard solution (RF value of 0.5). ASSAY PROCEDURE Solution A: 0.302 mg/mL of tetrabutylammonium phosphate and 13.4 mg/mL of dibasic sodium phosphate in water, adjusted with phosphoric acid to a pH of 3.8 before final dilution Mobile phase: Acetonitrile and Solution A (21:29). Allow to stand for 1 h, and if necessary, readjust with phosphoric acid to a pH of 3.8. Diluent: Acetonitrile and 5 mM monobasic potassium phosphate (17:3) Standard solution: 250 g/mL of USP Carbenicillin Indanyl Sodium RS in Diluent [NOTEContains the equivalent of 222 g/mL of carbenicillin.] Sample stock solution: Equivalent to 222 g/mL of carbenicillin from NLT 20 powdered Tablets in Diluent [NOTESonication may be necessary to dissolve. Contains 250 g/mL of carbenicillin indanyl sodium.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 2 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H18N2O6S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) P 100 rU rS CS
Official Monographs / Carbenicillin 67

= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carbenicillin Indanyl Sodium RS, calculated on the anhydrous basis, in the Standard solution (g/mL) = nominal concentration of carbenicillin in the CU Sample solution (g/mL) P = assigned potency of USP Carbenicillin Indanyl Sodium RS (g/mg) Acceptance criteria: 90%120% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Detector: UV Analytical wavelengths: Maximum at 267 nm and minimum at 254 nm Standard solution: USP Carbenicillin Indanyl Sodium RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H18N2O6S equivalent dissolved in the portion of Tablets taken: Result = (AU/AS) x (CS/CU) x (Mr1/Mr2) x 100 = absorbance of A267 A254 for the Sample solution = absorbance of A267 A254 for the Standard solution = concentration of USP Carbenicillin Indanyl CS Sodium RS in the Standard solution (g/mL) = nominal concentration of carbenicillin in the CU Sample solution (g/mL) = molecular weight of carbenicillin, 378.40 Mr1 = molecular weight of carbenicillin indanyl sodium, Mr2 516.54 Tolerances: NLT 75% (Q) of the labeled amount of C17H18N2O6S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements AU AS SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 2.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carbenicillin Indanyl Sodium RS
68
Carbidopa / Official Monographs

USP 32
Carbidopa
(Comment on this Monograph)id=m12810=Carbidopa=Ca-ChlMonos.pdf)
C10H14N2O4 H2O 244.24 Benzenepropanoic acid, -hydrazino-3,4-dihydroxy--methyl-, monohydrate, (S)-; (-)-L--Hydrazino-3,4-dihydroxy--methylhydrocinnamic acid monohydrate [38821-49-7]. Anhydrous 226.23 [28860-95-9]. DEFINITION Carbidopa contains NLT 98.0% and NMT 102.0% of C10H14N2O4 H2O. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Alcohol and 0.05 M monobasic sodium phosphate, adjusted with phosphoric acid to a pH of 2.7 (1:19) System suitability solution: 0.1 mg/mL of USP Carbidopa RS and 0.1 mg/mL of USP Methyldopa RS in Mobile phase Standard solution: 0.5 mg/mL of USP Carbidopa RS in Mobile phase [NOTEUse gentle heat and ultrasonification, if necessary, to dissolve.] Sample solution: 0.5 mg/mL of Carbidopa in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for methyldopa and carbidopa are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 0.9 between methyldopa and carbidopa in the System suitability solution Relative standard deviation: NMT 1.5% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H14N2O4 H2O in the portion of Carbidopa taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carbidopa RS, as the monohydrate, in the Standard solution (mg/mL) = concentration of Carbidopa in the Sample solution (mg/mL)
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE: LIMIT OF METHYLDOPA AND CARBIDOPA RELATED COMPOUND A Mobile phase, System suitability solution, Standard solution, Sample solution, Chromatographic system, and Suitability requirements: Proceed as directed in the Assay. Impurity standard solution: 2.5 g/mL of USP Methyldopa RS and 2.5 g/mL of USP Carbidopa Related Compound A RS in Mobile phase System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for methyldopa, carbidopa, and carbidopa related compound A are about 0.8, 1.0, and 1.8, respectively.] Analysis Samples: Sample solution and Impurity standard solution Calculate the percentage of methyldopa in the portion of Carbidopa taken: Result = (rU/rS) (CS/CU) 100 = peak response of methyldopa or carbidopa related compound A from the Sample solution rS = peak response of methyldopa or carbidopa related compound A from the Impurity standard solution = concentration of USP Methyldopa RS or USP CS Carbidopa Related Compound A RS in the Impurity standard solution (g/mL) = concentration of the Sample solution (g/mL) CU Acceptance criteria: NMT 0.5% of methyldopa, and NMT 0.5% of carbidopa related compound A rU SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 21.0 to 23.5, calculated as the monohydrate Sample solution: 10 mg/mL, in 666.6 mg/mL of aluminum chloride solution that has been filtered and adjusted with 0.25 N sodium hydroxide to a pH of 1.5 LOSS ON DRYING 731 Analysis: Heat 1 g in a suitable vacuum drying apparatus at 100 and a pressure of NMT 5 mm of mercury to constant weight. Cool and weigh. Acceptance criteria: It loses 6.9%7.9% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Carbidopa RS USP Carbidopa Related Compound A RS USP Methyldopa RS
Carbidopa and Levodopa Tablets

(Comment on this Monograph)id=m12870=Carbidopa and Levodopa Tablets=Ca-Chl-Monos.pdf) DEFINITION Carbidopa and Levodopa Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of carbidopa (C10H14N2O4) and levodopa (C9H11NO4).
USP 32
IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution A: 0.1 mg/mL of USP Carbidopa RS in 0.05 N hydrochloric acid and methanol (1:1) Standard solution B: 0.1 mg/mL of USP Levodopa RS in 0.05 N hydrochloric acid and methanol (1:1) Sample solution: Equivalent to 10 mg of carbidopa, from powdered Tablets, in a 100-mL volumetric flask containing 50 mL of 0.05 N hydrochloric acid. Agitate for 20 min, add methanol to volume, and filter or centrifuge. Application volume: 20 L Developing solvent system: Acetone, chloroform, nbutanol, glacial acetic acid, and water (12:8:8:8:7) Spray reagent: 3 mg/mL of ninhydrin in n-butanol and glacial acetic acid (97:3) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Develop, using the Developing solvent system, until the solvent front has moved 15 cm. Air-dry, spray uniformly with 0.5 mL of Spray reagent, and heat at 105 for 10 min. Acceptance criteria: The Sample solution exhibits two spots (reddish brown for levodopa and yellow-orange for carbidopa) having RF values that correspond to those exhibited by the Standard solutions. ASSAY PROCEDURE Solution A: 0.24 mg/mL of sodium 1-decanesulfonate in water Mobile phase: 11.04 g of monobasic sodium phosphate and 950 mL of water in a beaker. Add 1.3 mL of Solution A, and adjust with phosphoric acid to a pH of 2.8. Transfer to a 1-L volumetric flask, and dilute with water to volume. Standard solution: 50 mg of USP Levodopa RS in a 100mL volumetric flask. Add a quantity of USP Carbidopa RS, which is in a ratio with USP Levodopa RS that corresponds to the ratio of carbidopa to levodopa in the Tablets. Add 10 mL of 0.1 N phosphoric acid. Warm gently to dissolve the standards, and dilute with water to volume. Sample solution: Equivalent to 50 mg of levodopa from powdered NLT 20 Tablets, in a 100-mL volumetric flask. Add 10 mL of 0.1 N phosphoric acid, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min [NOTEAdjust until the retention times for levodopa and carbidopa are 4 min and 11 min, respectively.] Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 6 between levodopa and carbidopa Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H14N2O4 and C9H11NO4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response of carbidopa or levodopa from the Sample solution = peak response of carbidopa or levodopa from the Standard solution CS
Official Monographs / Carbinoxamine 69

= concentration of USP Carbidopa RS or USP Levodopa RS in the Standard solution (mg/mL) = nominal concentration of carbidopa or levodopa CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 750 mL Apparatus 1: 50 rpm Time: 30 min Analysis: Determine the amounts of carbidopa and levodopa in filtered portions of the solution under test, in comparison with a Standard solution having known concentrations of USP Carbidopa RS and USP Levodopa RS in Medium, as directed in the Assay. Tolerances: NLT 80% (Q) of the labeled amounts of C10H14N2O4 and C9H11NO4 are dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Carbidopa RS USP Levodopa RS
Carbinoxamine Maleate
(Comment on this Monograph)id=m12900=Carbinoxamine Maleate=Ca-Chl-Monos.pdf)
406.86 C16H19ClN2O C4H4O4 Ethanamine, 2-[(4-chlorophenyl)-2-pyridinylmethoxy]-N, Ndimethyl-, (Z)-2-butenedioate (1:1); 2-[p-Chloro--[2-(dimethylamino)ethoxy]benzyl]pyridine maleate (1:1) [3505-38-2]. DEFINITION Carbinoxamine Maleate contains NLT 98.0% and NMT 102.0% of C16H19ClN2O C4H4O4, on the previously dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 260 nm Sample solution: 50 g/mL in methanol Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0%. ASSAY PROCEDURE Sample solution: 8 mg/mL of Carbinoxamine Maleate, previously dried in glacial acetic acid Analysis: To 50 mL of Sample solution, add 1 drop of crystal violet TS, and titrate with 0.1 N perchloric acid VS to a bluegreen endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 20.34 mg of C16H19ClN2O C4H4O4.
70
Carbinoxamine / Official Monographs

98.0%102.0% Acceptance criteria: 93.0%107.0%
USP 32
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Standard solution: Chloroform Sample solution: Chloroform Eluant: Cyclohexane, chloroform, and diethylamine (15:3:2) Visualization: 1 SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 116121, determined after drying PH 791: 4.65.1, in a 10-mg/mL solution LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Carbinoxamine Maleate RS
Carbinoxamine Maleate Tablets

(Comment on this Monograph)id=m12920=Carbinoxamine Maleate Tablets=Ca-Chl-Monos.pdf) DEFINITION Carbinoxamine Maleate Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of carbinoxamine maleate (C16H19ClN2O C4H4O4). IDENTIFICATION ULTRAVIOLET ABSORPTION 197U Standard solution: 0.02 mg/mL of USP Carbinoxamine Maleate RS in 0.25 M sulfuric acid Sample solution: Prepare a solution equivalent to 0.02 mg/mL of carbinoxamine maleate in 0.25 M sulfuric acid, from the Tablets, as directed under Salts of Organic Nitrogenous Bases 501. Analytical wavelength: Peak maximum at 263 2 nm Acceptance criteria: The absorptivity of the Sample solution is within 7.0% of that of the Standard solution. ASSAY PROCEDURE Sample solution: Equivalent to 100 mg of carbinoxamine maleate from NLT 30 powdered Tablets. In a separator, add 35 mL of water and 3 g of sodium bicarbonate. Extract with five 20-mL portions of chloroform, filtering the extracts through a pledget of cotton. Evaporate the combined chloroform extracts on a steam bath just to dryness, dissolve the residue in 50 mL of glacial acetic acid, and add 1 drop of crystal violet TS. Analysis: Titrate with 0.05 N perchloric acid VS to a bluegreen endpoint. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.05 N perchloric acid is equivalent to 10.17 mg of C16H19ClN2O C4H4O4.
PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Detector: UV 260 nm Standard solution: USP Carbinoxamine Maleate RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution. Tolerances: NLT 75% (Q) of the labeled amount of C16H19ClN2O C4H4O4 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Standard solution: 40 g/mL of USP Carbinoxamine Maleate RS in methanol and water (9:1) Sample stock solution: Place 1 Tablet in a 100-mL volumetric flask, add 10.0 mL of water, and shake by mechanical means for 15 min. Dilute with methanol to volume, and filter, discarding the first 20 mL of the filtrate. Sample solution: Equivalent to 40 g/mL of carbinoxamine maleate from the Sample stock solution in methanol and water (9:1) Blank: Methanol and water (9:1) Spectrometric conditions Mode: UV-Vis Cell: 1 cm Analytical wavelength: Peak maximum at about 260 nm Analysis Samples: Standard solution, Sample solution, and Blank Calculate the percentage of C16H19ClN2O C4H4O4 in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Carbinoxamine Maleate RS in the Standard solution (g/mL) = nominal concentration of carbinoxamine maleate in the Sample solution (g/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Carbinoxamine Maleate RS
Carbol-Fuchsin Topical Solution

(Comment on this Monograph)id=m12980=Carbol-Fuchsin Topical Solution=Ca-Chl-Monos.pdf) Prepare Carbol-Fuchsin Topical Solution as follows:
Basic Fuchsin Phenol 3g 45 g
USP 32
Resorcinol Acetone Alcohol Purified Water To make 100 g 50 mL 100 mL A sufficient quantity 100 mL
Official Monographs / Carbon 71

hydroxide solution to fill the pipet and capillary connection up to the stopcock. Fill the buret with the leveling water, and draw it through the other stopcock opening in such a manner that all gas bubbles are eliminated from the system. Draw the Sample into the buret. By raising the leveling bottle, force the measured specimen into the pipet. The absorption may be facilitated by rocking the pipet or by flowing the specimen between pipet and buret. Draw any residual gas into the buret, and measure its volume. Acceptance criteria: NMT 1.0 mL of gas remains (NLT 99.0%, by volume, of CO2). IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF AMMONIA Sample: 1050 50 mL of the gas obtained as directed in the test for Nitrogen Dioxide (below) Analysis: Proceed with Carbon Dioxide as directed in the test for Nitrogen Dioxide (below), except to pass 1050 50 mL of this gas through an ammonia detector tube at the rate specified for the tube. Acceptance criteria: The indicator change corresponds to NMT 0.0025%. PROCEDURE 2: LIMIT OF HYDROGEN SULFIDE Sample: 1050 50 mL, released from the vapor phase Analysis: Pass 1050 50 mL, released from the vapor phase, through a hydrogen sulfide detector tube at the rate specified for the tube. Acceptance criteria: The indicator change corresponds to NMT 1 ppm. PROCEDURE 3: LIMIT OF NITRIC OXIDE Sample: 550 50 mL, released from the vapor phase Analysis: Pass 550 50 mL, released from the vapor phase, through a nitric oxidenitrogen dioxide detector tube at the rate specified for the tube. Acceptance criteria: The indicator change corresponds to NMT 2.5 ppm. PROCEDURE 4: CARBON MONOXIDE Sample: 1050 50 mL, released from the vapor phase of the contents of the container Analysis: Pass 1050 50 mL, released from the vapor phase of the contents of the container, through a carbon monoxide detector tube at the rate specified for the tube. Acceptance criteria: The indicator change corresponds to NMT 0.001%. PROCEDURE 5: NITROGEN DIOXIDE Sample: 550 50 mL, obtained as directed in the Analysis Analysis: Arrange the container so that when its valve is opened, a portion of the liquid phase of the contents is released through a piece of tubing of sufficient length to allow all of the liquid to vaporize during passage through it, and to prevent frost from reaching the inlet of the detector tube. Release into the tubing a flow of liquid sufficient to provide 550 mL of the vaporized specimen plus any excess necessary to ensure adequate flushing of air from the system. Pass 550 50 mL of this gas through a nitric oxide-nitrogen dioxide detector tube at the rate specified for the tube. Acceptance criteria: The indicator change corresponds to NMT 2.5 ppm. PROCEDURE 6: SULFUR DIOXIDE Sample: 1050 50 mL, obtained as directed in the test for Nitrogen Dioxide (above) Analysis: Proceed with Carbon Dioxide as directed in the test for Nitrogen Dioxide (above), except to pass 1050 50 mL through a sulfur dioxide detector tube at the rate specified for the tube. Acceptance criteria: The indicator change corresponds to NMT 5 ppm.
Dissolve Basic Fuchsin in a mixture of Acetone and Alcohol, and add to this solution Phenol and Resorcinol, previously dissolved in 725 mL of Purified Water. Then add sufficient Purified Water to make the product measure 1000 mL, and mix. OTHER COMPONENTS ALCOHOL DETERMINATION 611: SPECIFIC TESTS SPECIFIC GRAVITY 841: 7.0%10.0%
0.9901.050
Carbon Dioxide
(Comment on this Monograph)id=m13040=Carbon Dioxide=Ca-Chl-Monos.pdf) CO2 Carbon dioxide; Carbon dioxide [124-38-9]. 44.01
DEFINITION Carbon Dioxide contains NLT 99.0%, by volume, of CO2. [NOTEThe following tests are designed to reflect the quality of Carbon Dioxide in both its vapor and liquid phases, which are present in previously unopened cylinders. Reduce the container pressure by means of a regulator. Withdraw the specimens for the tests with the least possible release of Carbon Dioxide consistent with proper purging of the sampling apparatus. Measure the gases with a gas volume meter downstream from the detector tubes in order to minimize contamination or change of the specimens. Perform tests in the sequence in which they are listed.] The various detector tubes called for in the respective tests are listed in Reagents, Indicators, and SolutionsReagent Specifications. IDENTIFICATION PROCEDURE Sample: 100 5 mL, released from the vapor phase of the contents of the container Analysis: Pass the Sample through a Carbon Dioxide detector tube at the rate specified for the tube. Acceptance criteria: The indicator change extends throughout the entire indicating range of the tube. ASSAY PROCEDURE [NOTESampling for this Assay may be done from the vapor phase for convenience, but this method results in more residual volume. If the specification of 1 mL is exceeded from the vapor phase, a liquid specimen may be taken.] Sample: 100.0 mL of specimen taken from the liquid phase, as directed in the test for Nitrogen Dioxide (below) Analysis: Assemble a 100-mL gas buret, provided with a leveling bulb and two-way stopcock, and a gas absorption pipet of suitable capacity by connecting the pipet to one of the buret outlets. Fill the buret with slightly acidified water (turned pink with methyl orange), and fill the pipet with potassium hydroxide solution (1 in 2). By manipulation of the leveling bulb and leveling water, draw the potassium
72
Carbon / Official Monographs
USP 32
column containing support S3 and of sufficient length to separate CO2 from N2 and CO (which co-elute) and CO2 from NO2 at room temperature. A simple radioactivity detector and a thermal conductivity detector (or equivalent) are required for the mass determination of carbon monoxide. The radiochemical purity is NLT 98%. Mass analysis of the gasair mixture must demonstrate levels of carbon monoxide less than 1.23 mmoles in the entire dose, which is the upper limit for a single-bolus inhalation. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Dispense the gas either continuously or batchwise, and preserve in a single-dose container that is adequately shielded. It may also be trapped either on activated charcoal at 196 or on a molecular sieve at 72. LABELING: The label must include the following: the time and date of calibration; the amount of 11C as carbon monoxide expressed as total MBq (mCi) at time of calibration; the expiration time and date; and the statement Radioactive Material. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay, and states that the radioactive half-life of 11C is 20.39 min. Each container to hold 11CO shall be independently labeled to indicate lot number and/or batch number. The labeling states that a microbiological filter (0.22 m) is to be in place to remove any possible particulate matter that could be carried through to the final product.
SPECIFIC TESTS WATER DETERMINATION, Method I 921 Analysis: Flush the regulator that has been flushed with 5 L or more of the gas specimen. Pass 50 5 L, released from the vapor phase, through a water vapor detector tube connected to the regulator with a minimum length of metal or polyethylene tubing. Measure the gas passing through the detector tube with a gas flowmeter set at a flow rate of 2 L/min. Acceptance criteria: The indicator change corresponds to NMT 150 mg/cubic meter. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in cylinders.
Carbon Monoxide C 11
(Comment on this Monograph)id=m13050=Carbon Monoxide C 11=Ca-Chl-Monos.pdf) DEFINITION Carbon Monoxide C 11 is a colorless, odorless, nonirritating gas, suitable for administration by inhalation, in which a portion of the molecules are labeled with radioactive 11C. It contains NLT 90.0% and NMT 110.0% of the labeled amount of 11C expressed in MBq (or in mCi) at the time indicated in the labeling. IDENTIFICATION RADIONUCLIDE IDENTIFICATION (See Radioactivity 821.) A. Its gamma-ray spectrum is identical to that of a specimen of 11C in that it exhibits a positron annihilation peak at 0.511 MeV and possibly a sum peak of 1.022 MeV, dependent upon geometry and detector efficiency. B. Radio-gas chromatography, using a molecular sieve chromatographic column, to determine the absence of [13N]N2, using a suitable radioactivity detector and mass detector ASSAY PROCEDURE Analysis: Determine the radioactivity, in MBq (or mCi), by use of a calibrated system as directed under Radioactivity 821. Acceptance criteria: 90.0%110.0% SPECIFIC TESTS SPECIFIC ACTIVITY: Dependent upon the amount of radioactivity to be inhaled, but NMT 1.23 mmoles of carbon monoxide/volume RADIONUCLIDIC PURITY: A multichannel analyzer is used to count all radioactivity from 40 to 2,500 KeV to determine the absence of radiation, other than at 0.511 MeV and 1.022 MeV, over a period of 4 h. Possible impurities could be 13N2 [t1/2 = 9.97 min; this gamma-ray spectrum is indistinguishable from 11C, (B+) 491 KeV],10C [t1/2 = 19 s, 718.3 KeV (100%)], 14O [t1/2 = 70 s, 2312.7 KeV (99.4%)]. [11C]CO should contain NMT 10% impurities at the time of inhalation. RADIOCHEMICAL PURITY AND MASS DETERMINATION: [NOTE This pharmaceutical may be synthesized by different methods and may therefore contain different impurities. Additional validated tests relevant to the synthetic procedure may be necessary in order to ensure radiochemical purity of the final product.] Confirm by radio-gas chromatography. The gas stream, either directly from the target, or after initial chemical processing, is directed to an injection loop valve of a gas chromatograph and two precalibrated columns, a molecular sieve that allows separation of carbon monoxide from the different air components (O2, N2, and CH4), and a
Flumazenil C 11 Injection
(Comment on this Monograph)id=m13053=Flumazenil C 11 Injection=Ca-Chl-Monos.pdf) DEFINITION Flumazenil C 11 Injection is a sterile solution, suitable for intravenous administration, of Flumazenil in which a portion of the molecules are labeled at the N-position with radioactive 11C. It contains NLT 90.0% and NMT 110% of the labeled amount of 11C, expressed in MBq (or mCi) at the time indicated in the labeling. Its specific activity is NLT 14.8 GBq (400 mCi)/mol. It may contain suitable buffers. IDENTIFICATION RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is identical to that of a specimen of 11C in that it exhibits a positron annihilation peak at 0.511 MeV and possibly a sum peak of 1.022 MeV, dependent upon geometry and detector efficiency (see Radioactivity 821). ASSAY RADIOACTIVITY (See Radioactivity 821, Selection of a Counting Assembly.) Analysis: Using a suitable counting assembly, determine the radioactivity, in MBq (or mCi)/mL, of Injection by use of a calibrated system. Acceptance criteria: 90.0%110% IMPURITIES Organic impurities PROCEDURE: CHEMICAL PURITY Solution A: 1.2 mg/mL of monobasic sodium phosphate in deionized distilled water Mobile phase: Acetonitrile and Solution A (3:7) System suitability stock solution: 1 mg/mL of USP Flumazenil RS in acetonitrile System suitability solution: 10 g/mL of USP Flumazenil RS from System suitability stock in Mobile phase Sample solution: 0.1 mL/mL of Injection in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 20 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: System suitability solution Suitability requirements Column efficiency: NLT 100 theoretical plates Tailing factor: NMT 1.1 Relative standard deviation: NMT 3.2% Analysis Sample: Sample solution Separately calculate the percentage of each impurity in the portion of Injection taken: Result = (rU/rS) 100 = peak response for each impurity rU = sum of the responses of all the peaks rS Acceptance criteria: NMT 0.2% of any individual impurity, and NMT 0.9% of total impurities SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP Endotoxin Unit/mL, in which V is the maximum recommended total dose, in mL, at the expiration date or time PH 791: 4.58.5 RADIONUCLIDIC PURITY: Using a multichannel analyzer, count all radioactivity from 40 to 2,500 keV to determine the absence of radiation, other than at 0.511 MeV and 1.022 MeV, over a period of 4 h. Determine the half-life (20 min) by a suitable detector system. RADIOCHEMICAL PURITY Mobile phase and System suitability solution: Prepare as directed in the test for Chemical Purity. Chromatographic system and System suitability: Proceed as directed in the test for Chemical Purity, except that the liquid chromatograph is also equipped with a suitable collimated radiation detector (see Radioactivity 821). Analysis: Inject 20 L of the Injection into the chromatograph, record the chromatogram, and measure responses for the major peaks. Acceptance criteria: The radioactivity under the main peak is NLT 98% of the total radioactivity measured. SPECIFIC ACTIVITY Mobile phase and System suitability solution: Proceed as directed in the test for Chemical Purity. Chromatographic system and System suitability: Proceed as directed in the test for Chemical Purity, except that the liquid chromatograph is also equipped with a suitable collimated radiation detector (see Radioactivity 821). Analysis: Calculate the specific activity, in MBq (or mCi)/mol, of Injection taken: Result = (Mr CR PR)/(C 100) = molecular weight of flumazenil, 303.29 = radioactivity content as determined in the Assay [MBq (or mCi)/mL] PR = radiochemical purity as determined in the test for Radiochemical Purity (%) C = concentration of flumazenil in the Injection as determined in the test for Chemical Purity (g/mL) Acceptance criteria: NLT 400 mCi/mol OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that the Injection may be distributed or dispensed prior to completion of Sterility Tests 71, the latter test being started on the day following final manufacture, Mr CR

and except that it is not subject to the recommendation on Volume in Container. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers that are adequately shielded. LABELING: Label it to include the following, in addition to the information specified for Labeling under Injections 1: the time and date of calibration; the amount of 11C as [Nmethyl-11C]Flumazenil, expressed as MBq (or mCi); the specific activity, expressed as MBq (or mCi)/mol; the concentration, expressed as MBq (or mCi)/mL, at the date and time of calibration; the expiration date and time; the lot or batch number; the name and quantity of any added preservative or stabilizer; and the statements, Radioactive Material and Do not use if cloudy or if it contains particulate matter. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay, and states that the radioactive half-life of 11C is 20 min. USP REFERENCE STANDARDS 11 USP Endotoxin RS USP Flumazenil RS
Mespiperone C 11 Injection
(Comment on this Monograph)id=m13057=Mespiperone C 11 Injection=Ca-Chl-Monos.pdf)
DEFINITION Mespiperone C 11 Injection is a sterile, isotonic solution, suitable for intravenous administration, of 3-N-[11C] methylspiperone in which a portion of the molecules are labeled with radioactive 11C. It contains NLT 90.0% and NMT 110.0% of the labeled amount of 11C expressed in GBq (or mCi) at the time indicated in the labeling. Its specific activity is NLT 18.5 GBq (500 mCi)/mol. IDENTIFICATION RADIONUCLIDIC IDENTIFICATION: Its gamma- ray spectrum is identical to that of a specimen of 11C in that it exhibits a positron annihilation peak at 0.511 MeV and possibly a sum peak of 1.022 MeV, dependent on geometry and detector efficiency. (See Radioactivity 821.) ASSAY ASSAY FOR RADIOACTIVITY 821 Analysis: Using a suitable counting assembly (see Radioactivity 821, Selection of a Counting Assembly), determine the radioactivity, in GBq (or mCi)/mL, of Injection by use of a calibrated system. Acceptance criteria: 90.0%110% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP Endotoxin Unit/mL of the Injection, in which V is the maximum recommended total dose, in mL, at the expiration time PH 791: 4.57 RADIONUCLIDIC PURITY: Using a multichannel analyzer, count all radioactivity from 40 to 2500 KeV to determine the absence of radiation, other than at 0.511 MeV and 1.022
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expressed as total GBq (or mCi) at time of calibration; the expiration time and date; the lot or batch number; and the statements, [CAUTIONRadioactive Material] and Do not use if cloudy or if it contains particulate matter. The labeling indicates that in making dosage calculations correction is to be made for radioactive decay, and states that the radioactive half-life of 11C is 20 min. USP REFERENCE STANDARDS 11 USP Endotoxin RS
MeV, over a period of 4 h. Determine the half-life (20 min) by a suitable detector system. CHEMICAL PURITY Mobile phase: Acetonitrile and 0.05 M monobasic potassium phosphate (7:3) Standard solution: 0.1 mg/mL of 3-methylspiperone hydrochloride in Mobile phase Sample solution: 0.1 mg/mL from a volume of Injection in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm, and a suitable radioactivity detector (see Radioactivity 821) Column: 3.9-mm 30-cm; packing L1 Flow rate: 0.8 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 100 theoretical plates Tailing factor: NMT 1.1 Relative standard deviation: NMT 3.2% Analysis Samples: Standard solution and Sample solution Separately calculate the percentage of each impurity in the portion of the Injection taken: Result = (ri/rs) 100 = peak response for each impurity ri = sum of the responses of all the peaks rs Acceptance criteria Individual impurities: NMT 0.2% Total impurities: NMT 0.9% RADIOCHEMICAL PURITY 821: Proceed as directed in the test for Chemical Purity, except that the liquid chromatograph is also equipped with a suitable collimated radioactivity detector. The radioactivity under the main peak is NLT 98% of the total radioactivity measured. SPECIFIC ACTIVITY Mobile phase and Standard solution: Proceed as directed in the test for Chemical Purity. Chromatographic system: Proceed as directed in the test for Chemical Purity. Analysis: Calculate the specific activity, in MBq (or mCi)/mol, of Mespiperone C 11 Injection taken: Result = 3.11 (Cr Pr)/C = radioactivity content, as determined in the Assay for Radioactivity [MBq (or mCi)/mL] Pr = radiochemical purity, as determined in the test for Radiochemical Purity (%) C = concentration of 3-methylspiperone in the Injection as determined in the test for Chemical Purity (g/mL) Acceptance criteria: NLT 18.5 GBq/mol (400 mCi/mol) OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that the Injection may be distributed or dispensed prior to completion of the test for Sterility, the latter test being started on the day following final manufacture, and except that it is not subject to the recommendation on Volume in Container. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers that are adequately shielded. LABELING: Label it to include the following: the time and date of calibration; the amount of 11C as methylspiperone Cr
Methionine C 11 Injection
(Comment on this Monograph)id=m13060=Methionine C 11 Injection=Ca-Chl-Monos.pdf) DEFINITION Methionine C 11 Injection is a sterile isotonic solution, suitable for intravenous administration of L[11C]methionine, in which a portion of the molecules are labeled with radioactive 11C. It contains NLT 90.0% and NMT 110.0% of the labeled amount of 11C expressed in MBq (or in mCi) at the time indicated in the labeling. It may contain preservatives and stabilizers. IDENTIFICATION RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is identical to that of a specimen of 11C in that it exhibits a positron annihilation peak at 0.511 MeV and possibly a sum peak of 1.022 MeV, dependent upon geometry and detector efficiency. (See Radioactivity 821.) ASSAY ASSAY FOR RADIOACTIVITY 821 Analysis: Using a suitable counting assembly, (see Selection of a Counting Assembly) determine the radioactivity (see Radioactivity 821), in GBq (Ci)/mL, of the Injection by use of a calibrated system. Acceptance criteria: 90.0%110% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP Endotoxin Unit/mL of the Injection, in which V is the maximum recommended total dose, in mL, at the expiration time PH 791: 6.08.0 RADIONUCLIDIC PURITY: Using a suitable gamma-ray spectrometer, determine the absence of radiation other than at 0.511 MeV, over a period of 20 min. Determine the halflife (20.41 min) by a suitable detector system. CHEMICAL PURITY Mobile phase: 0.008 M copper acetate and 0.017 M Lproline, adjust with 0.030 M sodium acetate to a pH of 5 Standard solution: 0.1 mg/mL of DL-methionine in Mobile phase Sample solution: 0.1 mg/mL from a volume of Injection in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 0.5 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for D-methionine and Lmethionine are 1.0 and 2.4, respectively.]
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Suitability requirements Resolution: NLT 1.5 between the D- and L-enantiomers Column efficiency: NLT 1400 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of D-methionine in the portion of Injection taken: Result = (rU/rS) 100 rU = peak response for D-methionine = sum of the responses of all the peaks rS Acceptance criteria: NMT 4% of D-enantiomer is found RADIOCHEMICAL PURITY 821: Proceed as directed under Chemical Purity. The radioactivity of the main peak is NLT 98% of the total radioactivity measured. SPECIFIC ACTIVITY: NLT 37.0 GBq (1.0 Ci)/mmol OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that the Injection may be distributed or dispensed prior to completion of Sterility Tests 71 and Bacterial Endotoxins Test 85, these tests being started on the day of final manufacture, and except that it is not subject to the recommendation of Volume in Container. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers that are adequately shielded. LABELING: Label it to include the following: the time and date of calibration; the amount of 11C as methionine expressed as total MBq (or mCi)/mL at time of calibration; the expiration time and date; the name and quantity of any added preservative or stabilizer; and the statement [CAUTIONRadioactive Material]. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay, and states that the radioactive half-life of 11C is 20.4 min. Each container to hold 11C methionine shall be independently labeled to indicate lot number and batch number. The labeling states that a microbiological filter (0.22 m) is to be in place to remove any possible particulate matter that could be carried through to the final product. USP REFERENCE STANDARDS 11 USP Endotoxin RS

IDENTIFICATION RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is identical to that of a specimen of 11C in that it exhibits a positron annihilation peak at 0.511 MeV and possibly a sum peak of 1.022 MeV, dependent upon geometry and detector efficiency (see Radioactivity 821). ASSAY ASSAY FOR RADIOACTIVITY 821 (See Radioactivity 821, Selection of a Counting Assembly.) Analysis: Using a suitable counting assembly, determine the radioactivity, in MBq (or mCi)/mL, of Injection by use of a calibrated system. Acceptance criteria: 90.0%110% SPECIFIC TESTS RADIONUCLIDIC PURITY: Using a multichannel analyzer, count all radioactivity from 402500 keV to determine the absence of radiation, other than at 0.511 MeV and 1.022 MeV, over a period of 4 h. Determine the half-life (20 min) by a suitable detector system. CHEMICAL PURITY Mobile phase: Add 840 L of phosphoric acid to 500 mL of deionized distilled water in a 1000-mL volumetric flask. Add 270 mL of acetonitrile, and dilute with deionized distilled water to volume. Standard stock solution: 1 mg/mL of raclopride (as the tartrate salt) in water. Dilute a volume of the resulting solution with Mobile phase to obtain a concentration of 10 g/mL. Sample solution: Transfer a volume of Injection equivalent to 37 MBq (1 mCi) of radioactivity from a volume of Injection, with 10 parts of Mobile phase. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L9 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for odesmethylraclopride and for raclopride are 0.75 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between acetate and carbonate Column efficiency: NLT 85 theoretical plates Relative standard deviation: NMT 10.0% Analysis Samples: Standard solution and Sample solution Measure the areas of the responses for the raclopride peaks. Calculate the concentration, in g/mL, of C15H20Cl2N2O3 in the portion of Injection taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of the Sample solution = peak response of the Standard solution rS CS = concentration in the Standard solution (g/mL) CU = concentration in the Sample solution (g/mL) Acceptance criteria: In the chromatogram of the Sample solution, the area of the peak with a retention time of 6 min (raclopride) is NLT 98% of the total area of all peaks. RADIOCHEMICAL PURITY Mobile phase: Prepare as directed in the test for Chemical Purity. Standard solution: Prepare as directed in the test for Chemical Purity.
Raclopride C 11 Injection
(Comment on this Monograph)id=m13070=Raclopride C 11 Injection=Ca-Chl-Monos.pdf)
C1411CH20Cl2N2O3 [97849-54-2].
346.24
DEFINITION Raclopride C 11 Injection is a sterile solution, suitable for intravenous administration, of raclopride, in which a portion of the molecules are labeled at the O-methyl position with radioactive 11C. It contains NLT 90.0% and NMT 110% of the labeled amount of 11C expressed in MBq (or mCi) at the time indicated in the labeling. Its specific activity is NLT 18.5 Gbq (500 mCi)/mol. It may contain suitable buffers.
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Chromatographic system: Proceed as directed in the test for Chemical Purity, except that the liquid chromatograph is also equipped with a suitable collimated radiation detector (see Radioactivity 821). Acceptance criteria: NLT 95% of the radioactivity under the main peak [NOTEIts retention time is within 10% of that obtained for the Standard solution, similarly chromatographed.] BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP Endotoxin Unit/mL, in which V is the maximum recommended total dose, in mL, at the expiration date or time PH 791: 4.57 SPECIFIC ACTIVITY Mobile phase and Standard solution: Proceed as directed in the test for Chemical Purity. Chromatographic system: Proceed as directed in the test for Radiochemical Purity. Analysis: Calculate the specific activity, in GBq (or mCi)/mol, of the Injection taken: Result = (Mr Cr Pr)/(100 C) = molecular weight of raclopride, 347 = radioactivity content, as determined in the Assay for Radioactivity [GBq (or mCi)/mL] = radiochemical purity, as determined in the test Pr for Radiochemical Purity (%) C = concentration of raclopride in the Injection as determined in the test for Chemical Purity (g/mL) Acceptance criteria: NLT 18.5 GBq/mol (500 mCi/mol) OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that the Injection may be distributed or dispensed prior to completion of the test for Sterility, the latter test being started on the day following final manufacture, and except that it is not subject to the recommendation on Volume in Container. Mr Cr ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers that are adequately shielded. LABELING: Label it to include the following, in addition to the information specified for Labeling under Injections 1: the time and date of calibration; the amount of 11C as [omethyl-11C]raclopride, expressed as total megabecquerels (or millicuries); the specific activity, expressed as megabecquerels (or millicuries)/mol; and the concentration, expressed as megabecquerels (or millicuries)/mL, at the date and time of calibration; the expiration date and time; the lot or batch number; the name and quantity of any added preservative or stabilizer; and the statements, [CAUTION Radioactive Material] and Do not use if cloudy or if it contains particulate matter. The labeling indicates that in making dosage calculations correction is to be made for radioactive decay, and states that the radioactive half-life of 11C is 20 min. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Sodium Acetate C 11 Injection

(Comment on this Monograph)id=m13080=Sodium Acetate C 11 Injection=Ca-Chl-Monos.pdf)
DEFINITION Sodium Acetate C 11 Injection is a sterile solution, suitable for intravenous administration, of Sodium Acetate in which a portion of the carboxyl molecules are labeled with radioactive 11C. It contains NLT 90.0% and NMT 110.0% of the labeled amount of 11C expressed in MBq (or in Ci or mCi) at the time indicated in the labeling. It may contain suitable buffers. IDENTIFICATION RADIOACTIVITY, Radionuclides Identification 821: Its gammaray spectrum is identical to that of a specimen of 11C in that it exhibits a positron annihilation peak at 0.511 MeV and possibly a sum peak of 1.022 MeV, depending upon geometry and detector efficiency. ASSAY PROCEDURE (See Radioactivity 821, Selection of a Counting Assembly.) Analysis: Using a suitable counting assembly, determine the radioactivity in MBq (mCi)/mL of the Injection by use of a calibrated system. Acceptance criteria: 90.0%110% of the labeled amount of 11C SPECIFIC TESTS RADIONUCLIDIC PURITY 821: Use a multichannel analyzer to count all radioactivity from 402,500 keV to determine the absence of radiation, other than at 0.511 MeV and 1.022 MeV, over a period of 4 h. Determine the half-life by a suitable detector system. CHEMICAL PURITY Mobile phase Add 14 mL of 0.5 N sulfuric acid to 500 mL of water in a 1000-mL volumetric flask. Add 100 mL of acetonitrile, and dilute with water to volume. Standard solution: 1 mg/mL of sodium acetate in water. Dilute a portion of the resulting solution with Mobile phase to obtain a concentration of 20 g/mL. Sample solution: Dilute a volume of Injection equivalent to 1 mCi of radioactivity with 10 parts of Mobile phase. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 210 nm Column: 7.8-mm 10-cm; packing L9 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: Standard solution [NOTEThe relative retention times for acetate and carbonate are about 0.8 and 1.0 min, respectively.] Suitability requirements Resolution: NLT 85 theoretical plates Relative standard deviation: NMT 10.0% Analysis Samples Standard solution and Sample solution Measure the responses for the acetate peaks. Calculate the concentration, in g/mL, of sodium acetate in the Injection: Result = (rU/rS) CS = peak response of acetate from the Sample solution = peak response of acetate from the System rS suitability solution (g/mL) = concentration of sodium acetate in the Standard CS solution (g/mL) RADIOCHEMICAL PURITY Mobile phase and Standard solution: Proceed as directed under Chemical Purity. Sample solution: Sodium Acetate C 11 Injection, undiluted Chromatographic system and System suitability: Proceed as directed under Chemical Purity except that the liquid chromatograph is also equipped with a suitable collimated radiation detector (see Radioactivity 821). Analysis Sample: Sample Solution (30 L) Measure the areas for the major peaks. Acceptance criteria: The radioactivity under the acetate C 11 peak is NLT 95% of the total area of all peaks observed. [NOTEIts retention time is within 10% of that obtained for the Standard solution, similarly chromatographed.] BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP Endotoxin Unit/mL, in which V is the maximum recommended total dose, in mL, at the expiration date or time PH 791: 4.58.5 SPECIFIC ACTIVITY: NLT 3.7 GBq (100 mCi)/mol OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that the Injection may be distributed or dispensed prior to completion of the test for Sterility, the latter test being started on the day following final manufacture, and except that it is not subject to the recommendation of Volume in Container. rU

USP REFERENCE STANDARDS 11 USP Endotoxin RS
Urea C 13
(Comment on this Monograph)id=m13083=Urea C 13=Ca-ChlMonos.pdf) DEFINITION Urea C 13 contains NLT 99.0% and NMT 100.5% of
13
CH4N2O.
ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, and water (89:10:1) Biuret stock solution: 0.03 mg/mL of biuret in Mobile phase System suitability solution: 25 mg of Urea in 1 mL Biuret stock solution, and dilute with Mobile phase to 10 mL Standard solution: 2 mg/mL of USP Urea C 12 RS in Mobile phase Sample solution: 2 mg/mL of Urea C 13 in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 200 nm Column: 4.6-mm 25-cm; 5-m packing L8 Flow rate: 0.8 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.5 between urea and biuret Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Measure the areas for the major peaks. Calculate the percentage of 13CH4N2O in the portion of Urea C 13 taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Urea C 12 RS in the Standard solution (mg/mL) = concentration of Urea C 13 in the Sample CU solution (mg/mL) = molecular weight of Urea C 13 Mr1 = molecular weight of USP Urea C 12 RS Mr2 Acceptance criteria: 99.0%100.5% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION: NMT 0.1% HEAVY METALS Analysis: Dissolve 1.0 g in 20 mL of water, and add 5 mL of 0.1 N hydrochloric acid. Acceptance criteria: NMT 20 ppm SPECIFIC TESTS LIMIT OF BIURET Standard solution: 0.033 mg/mL of biuret Sample solution: 33.3 mg/mL of Urea C 13 Analysis: To the Sample solution and to 3 mL of the Standard solution, add 3 mL of sodium hydroxide solution (10 in 100) and 3 drops of copper sulfate solution (0.5 in 100), and allow to stand for 5 min. Acceptance criteria: NMT 0.1% of biuret. Any reddish violet color in the Sample solution is not more intense than that from the Standard solution.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers that are adequately shielded. LABELING: Label it to include the following, in addition to the information specified in Injections 1, Labeling: the time and date of calibration; the amount of 11C as labeled sodium acetate expressed as total MBq (or mCi) and the concentration as megabecquerels/mL (or as millicuries/mL), on the date and time of calibration; the expiration date and time; the lot or batch number; the name and quantity of any added preservative or stabilizer; an indication on the labeling that states, Do not use if cloudy or if it contains particulate matter; and the statement [CAUTIONRadioactive Material.]. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay, and also indicates that the radioactive half-life of 11C is 20 min.
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ISOTOPIC PURITY Sample solution: 12 mg/mL of Urea C 13 in methanol Chromatographic system (See Chromatography 621 and Mass Spectrometry 736.) Mode: GC connected to a mass spectrometer [NOTEThe mass spectrometer is operated in a single-ion response mode. The electron energy is 70 eV.] Column: 0.25-mm 15-m capillary column coated with a 0.1-m film of phase G47 Temperature Injection port: 250 Detector: 200 Transfer line to the mass spectrometer: 265 Carrier gas: Helium Injection size: 1 L Analysis Sample: Sample solution Record the total ion chromatogram, and combine all of the mass spectra scans across the entire major peak. Record the peak intensities at mass-to-charge ratios of 60, 61, 62, and 63. Calculate the percentage of carbon that is C 13 in the portion of Urea C 13: Result = [(I61 + I63)/(I60 + I61 + I63)] 100 = relative peak intensity at mass-to-charge ratio of 60 = relative peak intensity at mass-to-charge ratio of I61 61 = relative peak intensity at mass-to-charge ratio of I63 63 Acceptance criteria: NLT 99% is found Calculate the percentage of oxygen that is O 18 in the portion of Urea C 13: I60 Result = [(I62 + I63)/(I60 + I61 + I62 + I63)] 100 = relative peak intensity at a mass-to-charge ratio of 62, and the other terms are as defined above Acceptance criteria: NMT 15% is found MELTING RANGE OR TEMPERATURE: 132135 ALCOHOL-INSOLUBLE MATTER Analysis: Dissolve 5.0 g in 50 mL of warm alcohol, and if any insoluble residue remains, filter the solution on a tared filter, wash the residue, and the filter with 20 mL of warm alcohol, and dry at 105 for 1 h. Acceptance criteria: The weight of the residue does not exceed 2 mg (0.04%). CHLORIDE AND SULFATE 221, Chloride: A 2.0-g portion shows no more chloride than corresponds to 0.20 mL of 0.020 N hydrochloric acid (0.007%). CHLORIDE AND SULFATE 221, Sulfate: A 2.0-g portion shows no more sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid (0.010%). OTHER REQUIREMENTS: It meets the requirements for Identification tests A and B under Urea. I62
Urea C 13 for Oral Solution

(Comment on this Monograph)id=m13085=Urea C 13 for Oral Solution=Ca-Chl-Monos.pdf) DEFINITION Urea C 13 for Oral Solution is a dry powder prepared from Urea C 13. It contains NLT 90.0% and NMT 110.0% of the labeled amount of urea C 13 (13CH4N2O). It contains no preservatives. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, and water (89:10:1) Biuret stock solution: 0.03 mg/mL of biuret in Mobile phase System suitability solution: 25 mg of Urea in 1 mL Biuret stock solution, and dilute with Mobile phase to 10 mL Standard solution: 2 mg/mL of USP Urea C 12 RS in Mobile phase Sample solution: 2 mg/mL of Urea C 13 in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 200 nm Column: 4.6-mm 25-cm; 5-m packing L8 Flow rate: 0.8 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.5 between urea and biuret Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Measure the areas for the major peaks. Calculate the percentage of 13CH4N2O in the portion of Urea C 13 for Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Urea C 12 RS in the Standard solution (mg/mL) = nominal concentration of Urea C 13 in the CU Sample solution (mg/mL) = molecular weight of Urea C 13 Mr1 = molecular weight of USP Urea C 12 RS Mr2 Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS: packaged solids Meets the requirements for
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers at room temperature. USP REFERENCE STANDARDS 11 USP Urea C 13 RS
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62 and TESTS FOR SPECIFIED MICROORGANISMS 62: The total aerobic microbial count does not exceed 1000 cfu/g, the total combined molds and yeasts count does not exceed 100 cfu/g, and it meets the requirements for the absence of Salmonella species and Eschericha coli.
USP 32
COMPLETENESS OF SOLUTION 641: Meets the requirements, a solution in carbon dioxide-free water containing 100 mg/mL being used OTHER REQUIREMENTS: It meets the requirements for Identification tests A and B under Urea. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in sterile, well-closed containers. LABELING: Label it to indicate that the solution is to be discarded if particulate matter is visible after reconstitution. [NOTEIt is to be reconstituted with Sterile Purified Water.]
Official Monographs / Carboplatin 79

Identification solution: 40 mg/mL of urea Application volume: 20 L of the Sample solution and 4 L of the Identification solution Developing solvent system: n-Butanol saturated with water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Analysis Samples: Sample solution and Identification solution Locate the spots on the plate by spraying with Ehrlichs reagent. Determine the radioactivity distribution with a suitable radiation detector (see Radioactivity 821), and obtain the RF value. Acceptance criteria: NLT 90% of the total radioactivity is in the 14C band, and the RF value of the principal spot from the Sample solution corresponds to that from the Identification solution. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. LABELING: Label it to include the following: the amount of 14C, expressed in MBq (or Ci)/Capsule at the time of calibration; the expiration date; the total radioactivity/container; and the statement, Caution Radioactive Material.
Urea C 14 Capsules
(Comment on this Monograph)id=m13090=Urea C 14 Capsules=Ca-Chl-Monos.pdf)
DEFINITION Urea C 14 Capsules contain 14CH4N2O in which a portion of the molecules are labeled with radioactive 14C to provide 0.04 MBq (or 1 Ci) of radioactivity per capsule. It contains NLT 90.0% and NMT 110.0% of the labeled amount of 14C expressed as MBq (or Ci). IDENTIFICATION RADIOACTIVITY, Identification and Assay of Radionuclides 821 A solution of 1 or more Capsules in 1 N hydrochloric acid when tested using a liquid scintillation counter produces beta emission having a 49 keV mean and a 156 keV max. ASSAY PROCEDURE Assay of Radioactivity 821 Analysis: Prepare a solution of 1 or more Capsules in 1 N hydrochloric acid. Using a liquid scintillation counter, determine the radioactivity, in MBq (or mCi)/mL by use of a calibrated system. Acceptance criteria: 90.0%110.0% of the labeled amount of 14C. PERFORMANCE TESTS DISSOLUTION 711 Medium: Simulated gastric fluid TS; 500 mL Apparatus 1: 50 rpm Time: 10 min Analysis: Determine the background levels of 14C with a 1mL portion of the solution under test using a liquid scintillation counter. Tolerances: NLT 80% (Q) of the labeled amount of 14C is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS RADIONUCLIDIC PURITY 821 Analysis: Determine the radionuclidic purity of a solution of 1 or more Capsules in water using a liquid scintillation counter. Acceptance criteria: NLT 99.9% of the radioactivity is present as C 14. RADIOCHEMICAL PURITY Adsorbent: 0.25-mm layer of chromatographic cellulose Sample solution: Open 2 Capsules and place them in a suitable container, and add 8 mL of methanol.
Carboplatin
(Comment on this Monograph)id=m13120=Carboplatin=CaChl-Monos.pdf)
C6H12N2O4Pt Platinum, diammine[1,1-cyclobutanedicarboxylato(2-)O,O]-, (SP-4-2); cis-Diammine(1,1-cyclobutanedicarboxylato)platinum [41575-94-4]. DEFINITION Carboplatin contains NLT 98.0% and NMT 102.0% of C6H12N2O4Pt, calculated on the anhydrous basis.
371.25
[CAUTIONGreat care should be taken in handling Carboplatin because it is a suspected carcinogen.] IDENTIFICATION INFRARED ABSORPTION 197K: Meets the requirements
ASSAY PROCEDURE Mobile phase: Acetonitrile and water (87:13) Standard solution: 1 mg/mL of USP Carboplatin RS [NOTEUse this solution within 2 h.] Sample solution: 1 mg/mL of Carboplatin [NOTEUse this solution within 2 h.] Chromatographic system (See Chromatography 621, System Suitability.)
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Carboplatin / Official Monographs
USP 32
Mobile phase: Acetonitrile, Solution A, and water (5:1:44) Standard solution A: 0.005 mg/mL of 1,1cyclobutanedicarboxylic acid in Mobile phase Standard solution B: 1 mg/mL of USP Carboplatin RS System suitability solution: Mix 1.0 mL of Standard solution A with 1.0 mL of Standard solution B. Sample solution: 1 mg/mL of Carboplatin in Mobile phase Chromatographic System (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 100 L System suitability Sample: System suitability solution Suitability requirements [NOTEThe relative retention times are 0.65 for carboplatin, and 1.0 for 1,1-cyclobutanedicarboxylic acid.] Resolution: NLT 2.5 between the carboplatin and 1,1cyclobutanedicarboxylic acid peaks Column efficiency: NLT 1500 theoretical plates, determined from the cyclobutanedicarboxylic acid peak Relative standard deviation: NMT 10% Analysis Samples: Standard solution and Sample solution Calculate the percentage of 1,1-cyclobutanedicarboxylic acid in the portion of Carboplatin taken: Result = (rU/rs) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of 1,1-cyclobutanedicarboxylic acid in the Standard solution (mg/mL) = concentration of carboplatin in the Sample CU solution (mg/mL) Acceptance criteria: NMT 0.5% PROCEDURE 2 Mobile phase, Chromatographic system, and Analysis: Proceed as directed in the Assay. Standard solution: 2.5 g/mL of USP Carboplatin RS Sample solution: Use the Sample solution, prepared under Assay. Analysis Samples: Standard solution and Sample solution Analysis: Separately inject 10 L of the Standard solution and the Sample solution into the chromatograph, record the chromatograms, and measure the peak responses. Acceptance criteria [NOTEThe sum of the peak responses, excluding the carboplatin and 1,1-cyclobutanedicarboxylic acid responses, from the Sample solution, is NMT 2 times the carboplatin response from the Standard solution, and no single peak response is greater than that of the carboplatin peak from the Standard solution.] Individual impurities: NMT 0.25% Total impurities: NMT 0.5% SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 5.07.0, in a 10 mg/mL solution WATER DETERMINATION, Method I 921: NMT 0.5%, using anhydrous formamide as the solvent TRANSMITTANCE Sample solution: 10 mg/mL of Carboplatin Analysis: Determine the percent transmittance in 1-cm cells at a wavelength of 440 nm, using water as the blank. rU rS CS
Mode: LC Detector: UV 230 nm Column: 4.0-mm 30-cm; packing L8 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: NLT 3.0 Column efficiency: NLT 2500 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 1.2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H12N2O4Pt in the portion of Carboplatin taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carboplatin RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% rU rS CS OTHER COMPONENTS PLATINUM CONTENT [NOTEThoroughly cleanse all glassware with nitric acid and rinse with water to prevent mirroring of platinum precipitate.] Sample solution: Transfer 0.25 g of Carboplatin to a 600mL beaker. Add 400 mL of water, and slowly dissolve by heating almost to the boiling point on a hot plate covered with an insulating pad, stirring frequently with a glass rod. Analysis: When the Sample solution is complete, remove the insulating pad, and boil for about 10 min. Remove the beaker from the hot plate, allow to cool for 1 min without stirring, and filter through quantitative, fine-porosity, smooth, dense, ashless filter paper, collecting the filtrate in a 600-mL beaker, and completing the transfer to the filter with hot water. Wash the filter with hot water. Place the beaker containing the combined filtrate and washings on a hot plate, and evaporate to a volume of about 300 mL. Place a glass stirring rod in the beaker, and heat the solution to boiling. Slowly add to the center of the beaker, by dropwise additions, 10.0 mL of hydrazine hydrate, 85%. [CautionHydrazine is toxic.] Add 2 drops of 10 N sodium hydroxide, boil for 10 min to coagulate the precipitate for ease of filtration, cool, and filter through quantitative, medium-porosity, smooth, ashless filter paper. Rinse the beaker with hot water, and pour the rinsings onto the filter. Wipe the beaker and the stirring rod with small pieces of the same kind of paper used for this filtration, and place these and the filter containing the precipitate in a No. 1 porcelain crucible, previously ignited to constant weight. Dry on a hot plate covered with an insulating pad, slowly increase the heat to char, and ignite for 1 h at 800. Cool in a desiccator, and weigh again. Acceptance criteria: The weight of the platinum so obtained is between 52.0%53.0% of the carboplatin taken. IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF 1,1-CYCLOBUTANEDICARBOXYLIC ACID Solution A: Dissolve 8.5 g of tetrabutylammonium hydrogen sulfate in 80 mL of water. Add 3.4 mL of phosphoric acid, and adjust with 10 N sodium hydroxide to a pH of 7.55.
USP 32
Acceptance criteria: NLT 97% transmittance WATER-INSOLUBLE MATTER Sample solution: Dissolve 1 g of carboplatin in 100 mL of water. Dissolve by stirring with a stirring bar for 30 min. With the aid of suction, filter through a tared filtering crucible. Rinse the beaker with water, and transfer the rinsings to the crucible. Dry the crucible at 130 10 to constant weight. Acceptance criteria: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. USP REFERENCE STANDARDS 11 USP Carboplatin RS
Official Monographs / Carboplatin 81

Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4.0-mm 30-cm; packing L8 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: NLT 3.0 Column efficiency: NLT 2500 theoretical plates Tailing factor: NMT 2.5 Relative standard deviation: NMT 1.2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H12N2O4Pt in the portion of Carboplatin for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carboplatin RS in the Standard solution (mg/mL) = nominal concentration of carboplatin in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Carboplatin for Injection

(Comment on this Monograph)id=m13125=Carboplatin for Injection=Ca-Chl-Monos.pdf) DEFINITION Carboplatin for Injection is a sterile, lyophilized mixture of Carboplatin and Mannitol. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C6H12N2O4Pt. [CAUTIONGreat care should be taken in handling Carboplatin because it is a suspected carcinogen.] IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 621 Standard solution: 10 mg/mL of USP Carboplatin RS in water Sample solution: 10 mg/mL of carboplatin, from contents of 1 container in water Chromatographic system Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 10 L Developing solvent system: Acetone and water (4:1) Spray reagent: Add 5.6 g of stannous chloride to 10 mL of hydrochloric acid, and stir for 5 min. [NOTEIt is not necessary that all of the solids dissolve.] Add 90 mL of water and 1 g of potassium iodide, and stir. [NOTEPrepare this solution fresh daily.] Analysis Samples: Standard solution and Sample solution Place the plate in a chromatographic chamber lined with filter paper and equilibrated for 2 h in Developing solvent system. Develop the chromatogram until the solvent front has moved 10 cm from the origin. Remove the plate from the chamber, and air-dry at room temperature for 2 h. Spray with the Spray reagent, and heat at 110 for 10 min. Acceptance criteria: The principal spot from the Sample solution corresponds in appearance and RF value to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (87:13) Standard solution: 1 mg/mL of USP Carboplatin RS [NOTEUse this solution within 2 h.] Sample solution: 1 mg/mL carboplatin, from the contents of 1 container diluted with water [NOTEComplete chromatographic analysis of this solution within 2 h.]
IMPURITIES Organic Impurities PROCEDURE: LIMIT OF 1,1-CYCLOBUTANEDICARBOXYLIC ACID Solution A: Dissolve 8.5 g of tetrabutylammonium hydrogen sulfate in 80 mL of water. Add 3.4 mL of phosphoric acid, and adjust with 10 N sodium hydroxide to a pH of 7.55. Mobile phase: Acetonitrile, Solution A, and water (5:44:1) Standard solution: 0.01 mg/mL of 1,1cyclobutanedicarboxylic acid in Mobile phase System suitability solution: Mix 1.0 mL of the Standard solution with 1.0 mL of Standard solution from the Assay. Sample solution: 1 mg/mL carboplatin, from the contents of 1 container diluted with Mobile phase [NOTEComplete the chromatographic analysis of the solution within 2 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 100 L System suitability Sample: System suitability solution [NOTEThe relative retention times for carboplatin and 1,1-cyclobutanedicarboxylic acid are about 0.65 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between carboplatin and 1,1cyclobutanedicarboxylic acid peaks Column efficiency: NLT 1500 theoretical plates for the cyclobutanedicarboxylic acid peak
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Carboplatin / Official Monographs

Relative standard deviation: NMT 10.0% Analysis Sample: Standard solution and Sample solution Calculate the percentage of 1,1-cyclobutanedicarboxylic acid in the portion of Carboplatin for Injection: Result = (rU/rS) (CS/CU) 100 = peak response for 1,1-cyclobutanedicarboxylic acid from the Sample solution = peak response for 1,1-cyclobutanedicarboxylic rS acid from the Standard solution CS = concentration of 1,1-cyclobutanedicarboxylic acid in the Standard solution (mg/mL) CU = nominal concentration of carboplatin in the Sample solution (mg/mL) Acceptance criteria: NMT 1.0% rU
USP 32
Carboprost Tromethamine
(Comment on this Monograph)id=m13140=Carboprost Tromethamine=Ca-Chl-Monos.pdf)
SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: 0.54 USP Endotoxin Unit/mg of carboplatin STERILITY TESTS 71: Meets the requirements when tested as directed in Test for Sterility of the Product to Be Examined, Membrane Filtration CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. PH 791: 5.07.0, in a solution constituted as directed in the labeling, Sterile Water for Injection being used WATER DETERMINATION, Method I 921 Analysis: Use anhydrous formamide as the extraction solvent. Introduce 50 mL of anhydrous formamide into the titration vessel, and titrate with the reagent to the electrometric endpoint. Use the formamide thus dried to rinse a suitable glass syringe equipped with a 22-gauge needle, 8 cm long. Add the rinse back to the titration vessel, and titrate the vessel contents again, if necessary. Via the syringe, withdraw 5 mL of the formamide thus titrated and, through the closure of the container, expel the contents into the container. Shake the container to obtain a solution. With the same syringe, withdraw all of the contents of the container, and transfer to the titration vessel. Titrate to the endpoint, adjusting the feeding speed control to the lowest setting to avoid overtitration. Acceptance criteria: NMT 3.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in Injections 1, Containers for Sterile Solids, and protect from light. USP REFERENCE STANDARDS 11 USP Carboplatin RS USP Endotoxin RS
C21H36O5 C4H11NO3 489.64 Prosta-5,13-dien-1-oic acid, 9,11,15-trihydroxy-15-methyl-, (5Z, 9,11,13E,15S)-, compound with 2-amino-2(hydroxymethyl)-1,3-propanediol (1:1); (Z)-7-[(1R,2R,3R,5S)-3,5-Dihydroxy-2-[(E)-(3S)-3-hydroxy-3methyl-1-octenyl]cyclopentyl]-5-heptenoic acid compound with 2-amino-2-(hydroxymethyl)-1,3-propanediol (1:1); (15S)-15-Methylprostaglandin F2 tromethamine [58551-69-2]. DEFINITION Carboprost Tromethamine contains NLT 95.0% and NMT 105.0% of C25H47NO8, calculated on the dried basis. [CAUTIONGreat care should be taken to prevent inhaling particles of Carboprost Tromethamine and exposing the skin to it.] IDENTIFICATION INFRARED ABSORPTION 197M ASSAY PROCEDURE Mobile phase: Methylene chloride, 1,3-butanediol, and water (992:7:0.5) Internal standard solution: 7 mg/mL of guaifenesin in Mobile phase Buffer solution: Dissolve 10.5 g of citric acid in 75 mL of water. Add 5 N sodium hydroxide slowly to adjust to a pH of 4.0, and dilute with water to 100 mL. Standard solution: Transfer 5 mg of USP Carboprost Tromethamine RS to a stoppered, 50-mL centrifuge tube. Add 20.0 mL of methylene chloride and 2 mL of Buffer solution. Shake the stoppered tube for 10 min, and centrifuge. Remove and discard the top (aqueous) layer, and transfer a 4.0-mL aliquot of the lower (methylene chloride) layer to a suitable vial. Evaporate with the aid of a stream of nitrogen to dryness. Add 100 L of a freshly prepared solution of -bromo-2-acetonaphthone in acetonitrile (1 in 50). Swirl to wash down the sides of the vial. Add 50 L of
USP 32
a freshly prepared solution of diisopropylethylamine in acetonitrile (1 in 100), swirl again, and place the vial in a suitable heating device maintained at a temperature of 30 to 35 for NLT 15 min. Evaporate the acetonitrile from the vial with the aid of a stream of nitrogen, add 2.0 mL of Internal standard solution, mix, and pass the resulting solution through a fine-porosity filter. Protect the filtered solution from light prior to injection to prevent degradation of the naphthacyl ester of carboprost. Sample solution: Proceed as directed for Standard solution, except to use Carboprost Tromethamine in place of USP Carboprost Tromethamine RS. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm stainless steel; 10-m packing L3 Flow rate: 1.8 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for guaifenesin and 2naphthacyl ester of carboprost are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between guaifenesin and the 2naphthacyl ester of carboprost Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C25H47NO8 in the portion of Carboprost Tromethamine taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the 2-naphthacyl ester of carboprost to the internal standard of the Sample solution RS = peak response ratio of the 2-naphthacyl ester of carboprost to the internal standard of the Standard solution CS = concentration of USP Carboprost Tromethamine RS in the Standard solution (mg/mL) CU = concentration of carboprost tromethamine in the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.5% Organic Impurities PROCEDURE: LIMIT OF 15R-EPIMER AND 5-trans ISOMER Buffer solution, Mobile phase, Internal standard solution, and Sample solution: Proceed as directed in the Assay. Chromatographic system: (See Chromatography 621, System Suitability.) Proceed as directed in the Assay, except use the following injection size: Injection size: 25 L System suitability: Proceed as directed in the Assay, using the Sample solution in place of the Standard solution. Analysis Sample: Sample solution Calculate the percentage of the 15R-epimer (as the tromethamine salt) in the portion of Carboprost Tromethamine taken: Result = rA/(rA + rB + rC) 100 rA = area of any peak at a relative retention time of 0.7 RU rB
Official Monographs / Carboprost 83

= area of any peak at a relative retention time of 1.0 = area of any peak at a relative retention time of rC 1.2 Calculate the percentage of the 5-trans isomer (as the tromethamine salt) in the portion of Carboprost Tromethamine taken: Result = rC/(rA + rB + rC) 100 = area of any peak at a relative retention time of 1.2 = area of any peak at a relative retention time of rA 0.7 = area of any peak at a relative retention time of rB 1.0 Acceptance criteria 15R-epimer: NMT 2.0% 5-trans isomer: NMT 3.0% rC SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +18 to +24 Sample solution: 10 mg/mL in alcohol LOSS ON DRYING 731: Dry it in a vacuum at a pressure not exceeding 5 mm of mercury at 50 for 16 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store in a freezer. USP REFERENCE STANDARDS 11 USP Carboprost Tromethamine RS
Carboprost Tromethamine Injection

(Comment on this Monograph)id=m13150=Carboprost Tromethamine Injection=Ca-Chl-Monos.pdf) DEFINITION Carboprost Tromethamine Injection is a sterile solution of Carboprost Tromethamine in aqueous solution, which may also contain Benzyl Alcohol, Sodium Chloride, and Tromethamine. It contains NLT 90.0% and NMT 110.0% of the labeled amount of carboprost (C21H36O5). IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Extract the equivalent of 2.5 mg of carboprost tromethamine from a volume of Injection, with 1.52 times its volume of chloroform. Discard the chloroform layer, and acidify the aqueous layer with 35 drops of hydrochloric acid. Extract the acidified solution with an equivalent volume of chloroform. Filter the chloroform layer through a pledget of cotton, and concentrate it to a volume of less than 1 mL. Combine the resulting solution with 150180 mg of potassium bromide. Dry the potassium bromide mixture in a vacuum overnight, and prepare a pellet from the dried mixture. ASSAY PROCEDURE Mobile phase: Methylene chloride, 1,3-butanediol, and water (992:7:0.5) Internal standard solution: 3 mg/mL of guaifenesin in Mobile phase Buffer solution: Dissolve 10.5 g of citric acid in 75 mL of water. Add 5 N sodium hydroxide slowly to adjust to a pH of 4.0, and dilute with water to 100 mL. Standard solution: Prepare a solution containing 0.332 mg/mL of USP Carboprost Tromethamine RS and 9 mg/mL of benzyl alcohol. Transfer 2.0 mL into a stoppered
84
Carboprost / Official Monographs
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, and store in a refrigerator. USP REFERENCE STANDARDS 11 USP Carboprost Tromethamine RS USP Endotoxin RS
centrifuge tube. Add 20.0 mL of methylene chloride and 1.0 mL of Buffer solution, shake the stoppered tube for 10 min, and centrifuge. Remove and discard the top (aqueous) layer, transfer an 8.0-mL aliquot of the lower (methylene chloride) layer to a suitable vial, and evaporate the solution with the aid of a stream of nitrogen. [NOTEThe residue does not evaporate to dryness because of the presence of benzyl alcohol.] Add 100 L of a freshly prepared solution of bromo-2-acetonaphthone in acetonitrile (1 in 50), and swirl to wash down the sides of the vial. Add 50 L of a freshly prepared solution of diisopropylethylamine in acetonitrile (1 in 100). Swirl again, and place the vial in a suitable heating device maintained at a temperature of 30 to 35 for NLT 15 min. Evaporate the acetonitrile from the vial with the aid of a stream of nitrogen, add 1.0 mL of Internal standard solution, mix, and pass the resulting solution through a fineporosity filter. Protect the filtered solution from light prior to injection to prevent degradation of the naphthacyl ester of carboprost. Sample solution: Pipet a volume of Injection, nominally equivalent to 500 g of carboprost, to a stoppered, 50-mL centrifuge tube. Proceed as directed for Standard solution, beginning with Add 20.0 mL of methylene chloride. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm stainless steel; 10-m packing L3 Flow rate: 1.8 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for guaifenesin and 2naphthacyl ester of carboprost are 0.6 and 1.0, respectively. ] Suitability requirements Resolution: NLT 4.0 between guaifenesin and the 2naphthacyl ester of carboprost Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H36O5 in each mL of Injection taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = peak response ratios of the 2-naphthacyl ester of carboprost to the internal standard of the Sample solution RS = peak response ratios of the 2-naphthacyl ester of carboprost to the internal standard of the Standard solution CS = concentration of USP Carboprost Tromethamine RS in the Standard solution (g/mL) CU = nominal concentration of carboprost in the Sample solution (mg/mL) Mr1 = molecular weight of carboprost, 368.51 Mr2 = molecular weight of carboprost tromethamine, 489.64 Acceptance criteria: 90.0%110.0% RU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 714.3 USP Endotoxin Units/mg of carboprost tromethamine PH 791: 7.08.0 OTHER REQUIREMENTS: Meets the requirements under Injections 1
Carboxymethylcellulose Sodium
(Comment on this Monograph)id=m13210=Carboxymethylcellulose Sodium=CaChl-Monos.pdf) Cellulose carboxymethyl ether sodium salt [9004-32-4]. DEFINITION Carboxymethylcellulose Sodium is the sodium salt of a polycarboxymethyl ether of cellulose. It contains NLT 6.5% and NMT 9.5% of sodium (Na), calculated on the dried basis. IDENTIFICATION A. PROCEDURE Sample solution: Add about 1 g of powdered Carboxymethylcellulose Sodium to 50 mL of water, while stirring to produce a uniform dispersion. Continue the stirring until a clear solution is produced. Analysis: To 1 mL of the Sample solution, diluted with an equal volume of water in a small test tube, add 5 drops of 1-naphthol TS. Incline the test tube, and carefully introduce down the side of the tube 2 mL of sulfuric acid so that it forms a lower layer. Acceptance criteria: A red-purple color develops at the interface. B. PROCEDURE Sample solution: Add 1 g of powdered Carboxymethylcellulose Sodium to 50 mL of water, while stirring to produce a uniform dispersion. Continue the stirring until a clear solution is produced. Analysis: To 5 mL of the Sample solution, add an equal volume of barium chloride TS. Acceptance criteria: A fine, white precipitate is formed. C. IDENTIFICATON TESTSGENERAL, Sodium 191: A portion of the Sample solution meets the requirements. Sample solution: Add 1 g of powdered Carboxymethylcellulose Sodium to 50 mL of water, while stirring to produce a uniform dispersion. Continue the stirring until a clear solution is produced. ASSAY PROCEDURE Sample solution: Transfer to a beaker 500 mg of Carboxymethylcellulose Sodium, add 80 mL of glacial acetic acid, heat the mixture on a boiling water bath for 2 h, and cool to room temperature. Analysis: Titrate the Sample solution with 0.1 N perchloric acid VS. Each mL of 0.1 N perchloric acid is equivalent to 2.299 mg of Na. Acceptance criteria: NLT 6.5% and NMT 9.5% of Na, calculated on the dried basis IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm, adding 1 mL of hydroxylamine hydrochloride solution (1 in 5) to the solution of the residue
USP 32
SPECIFIC TESTS VISCOSITY 911 Analysis: Determine the viscosity in a water solution at the concentration stated on the label. Using undried Carboxymethylcellulose Sodium, weigh the amount that, on the dried basis, will provide 200 g of solution of the stated concentration. Add the substance in small amounts to 180 mL of stirred water contained in a tared, wide-mouth bottle, continue stirring rapidly until the powder is well wetted, add sufficient water to make the mixture weigh 200 g, and allow to stand, with occasional stirring, until solution is complete. Adjust the temperature to 25 0.2, and determine the viscosity, using a rotational type of viscosimeter, making certain that the system reaches equilibrium before taking the final reading. Acceptance criteria: The viscosity of solutions of 2% or higher concentration is NLT 80.0% and NMT 120.0% of that stated on the label; the viscosity of solutions of less than 2% concentration is NLT 75.0% and NMT 140.0% of that stated on the label. PH 791: 6.58.5 in a solution (1 in 100) LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 10.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate the viscosity in solutions of stated concentrations.
Official Monographs / Carboxymethylcellulose 85

Analysis: Titrate the Sample solution with 0.1 N perchloric acid in dioxane VS, determining the endpoint potentiometrically. Each mL of 0.1 N perchloric acid is equivalent to 29.67 mg of carboxymethylcellulose sodium. Acceptance criteria: 16.0%17.0% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 50 ppm, using 400 mg of Carboxymethylcellulose Sodium Paste and adding 1 mL of hydroxylamine hydrochloride solution (1 in 5) to the solution of the residue SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: The total bacterial count does not exceed 1000 cfu/g, and the tests for Salmonella species and Escherichia coli are negative. CONSISTENCY Apparatus: Determine the consistency of Paste by means of a penetrometer fitted with a polished cone-shaped metal plunger weighing 150 g, having a detachable steel tip of the following dimensions: the tip of the cone has an angle of 30, the point being truncated to a diameter of 0.381 0.025 mm, the base of the tip is 8.38 0.05 mm in diameter, and the length of the tip is 14.94 0.05 mm. The remaining portion of the cone has an angle of 90, is 28 mm in height, and has a maximum diameter at the base of about 65 mm. The containers for the test are flat-bottom metal cylinders that are 100 6 mm in diameter and NLT 65 mm in height. They are constructed of at least 1.6-mm (16-gauge) metal, and are provided with well-fitting, watertight covers. Analysis: Place the required number of containers in an oven, and bring them and a quantity of Paste to a temperature of 82 2.5, pour the Paste into one or more of the containers, filling to within 6 mm of the rim. Cool to 25 2.5 over a period of NLT 16 h, protected from drafts. Two h before the test, place the containers in a water bath at 25 0.5. If the room temperature is below 23.5 or above 26.5, adjust the temperature of the cone to 25 0.5 by placing it in the water bath. Without disturbing the surface of the substance under test, place the container on the penetrometer table, and lower the cone until the tip just touches the top surface of the test substance at a spot 2538 mm from the edge of the container. Adjust the zero setting and quickly release the plunger, then hold it free for 5 s. Secure the plunger, and read the total penetration from the scale. Make three or more trials, each so spaced that there is no overlapping of the areas of penetration. Where the penetration exceeds 20 mm, use a separate container of the test substance for each trial. Read the penetration to the nearest 0.1 mm. Calculate the average of the three or more readings, and conduct further trials to a total of 10 if the individual results differ from the average by more than 3%. Acceptance criteria: The final average of the trials is NLT 30.0 mm and NMT 36.0 mm, indicating a consistency value between 300 and 360. LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and avoid prolonged exposure to temperatures exceeding 30.
Carboxymethylcellulose Sodium Paste

(Comment on this Monograph)id=m13240=Carboxymethylcellulose Sodium Paste=Ca-Chl-Monos.pdf) DEFINITION Carboxymethylcellulose Sodium Paste contains NLT 16.0% and NMT 17.0% of carboxymethylcellulose sodium. IDENTIFICATION A. PROCEDURE Sample solution: Digest a quantity of Paste equivalent to 1 g of carboxymethylcellulose sodium with 50 mL of water until the solution is virtually complete, and filter. Analysis: To 30 mL of the Sample solution, add 3 mL of hydrochloric acid. Acceptance criteria: A white precipitate is formed. [NOTE Filter the solution and save the filtrate for use in Identification test C.] B. PROCEDURE Sample solution: Use the remainder of the Sample solution prepared under Identification test A. Analysis: To the Sample solution add an equal volume of barium chloride TS. Acceptance criteria: A fine, white precipitate is formed. C. IDENTIFICATION TESTSGENERAL, Sodium 191: The filtrate from Identification test A responds to the tests. ASSAY PROCEDURE Sample solution: Transfer 2 g of Paste to a glass-stoppered, 250-mL conical flask. Add 75 mL of glacial acetic acid, attach a condenser, and reflux for 2 h. Cool, transfer the mixture to a 250-mL beaker with the aid of small volumes of glacial acetic acid.
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USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K B. The RF value of the principal spot due to carisoprodol of the Sample solution obtained in the test for Limit of Meprobamate corresponds to that obtained in a Standard solution of USP Carisoprodol RS in chloroform containing 100 mg/mL. ASSAY PROCEDURE Sodium methoxide titrant: Prepare and standardize as directed for 0.1 N sodium methoxide (in toluene) (see Reagents, Indicators, and SolutionsVolumetric Solutions), except to use 200 mL of methanol to dissolve the sodium metal, to add 750 mL of toluene, and to dilute with methanol to 1000 mL. Sample: 400 mg of Carisoprodol Analysis: Add 10 mL of pyridine to the Sample. Add 1 drop of phenolphthalein TS, and titrate with Sodium methoxide titrant to a permanent pink endpoint. Add 25.0 mL of Sodium methoxide titrant, connect the flask to a water-cooled condenser, and reflux on a hot plate for 30 min. Allow to cool, add 40 mL of alcohol and 7 drops of phenolphthalein TS. Titrate the excess alkali with 0.1 N hydrochloric acid VS until the pink color disappears. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of Sodium methoxide titrant is equivalent to 26.03 mg of C12H24N2O4. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE: LIMIT OF MEPROBAMATE Standard solution: 1 mg/mL of USP Meprobamate RS in chloroform Sample solution: 100 mg/mL of Carisoprodol in chloroform Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 10 L for Sample solution and 5 L for Standard solution Developing solvent system: Chloroform and acetone (4:1) Spray reagent 1: Antimony trichloride TS Spray reagent 2: 3 in 100 solution of furfural in chloroform Analysis Samples: Standard solution and Sample solution Proceed as directed in the chapter. Allow the spots to dry in a current of air, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the solvent to evaporate, and spray the plate alternately with Spray reagent 1 and Spray reagent 2 until one or more black spots appear, heat the plate at 110 for 15 min, and examine the plate. Acceptance criteria: Any spot in the Sample solution having an RF value corresponding to that of meprobamate in the Standard solution is not darker in color than the meprobamate spot in the Standard solution (NMT 0.5%).
Carboxymethylcellulose Sodium Tablets

(Comment on this Monograph)id=m13250=Carboxymethylcellulose Sodium Tablets=Ca-Chl-Monos.pdf) DEFINITION Carboxymethylcellulose Sodium Tablets contain an amount of sodium (Na) equivalent to NLT 6.5% and NMT 9.5% of the labeled amount of carboxymethylcellulose sodium. IDENTIFICATION A. PROCEDURE Sample solution: Digest a quantity of powdered Tablets equivalent to 1 g of carboxymethylcellulose sodium with 50 mL of water until the solution is virturally complete, and filter. Analysis: To 30 mL of the Sample solution, add 3 mL of hydrochloric acid. Acceptance criteria: A white precipitate is formed. [NOTE Filter the solution and save the filtrate for use in Identification test A.] B. PROCEDURE Sample solution: Use the remainder of the Sample solution prepared under Identification test A. Analysis: To the Sample solution add an equal volume of barium chloride TS. Acceptance criteria: A fine, white precipitate is formed. C. IDENTIFICATION TESTSGENERAL, Sodium 191: The filtrate from Identification test A meets the requirements. ASSAY PROCEDURE Sample solution: Dissolve an amount equivalent to 500 mg of carboxymethylcellulose sodium from powdered Tablets (NLT 20 Tablets) in 80 mL of glacial acetic acid. Heat the mixture on a steam bath for 2 h and cool to room temperature. Analysis: Titrate the Sample solution with 0.1 N perchloric acid VS. Each mL of 0.1 N perchloric acid is equivalent to 2.299 mg of Na. Acceptance criteria: 6.5%9.5% PERFORMANCE TESTS DISINTEGRATION 701 Time: 2 h UNIFORMITY OF DOSAGE UNITS 905:
Carisoprodol
(Comment on this Monograph)id=m13370=Carisoprodol=CaChl-Monos.pdf)
C12H24N2O4 ()-2-Methyl-2-propyl-1,3-propanediol carbamate isopropylcarbamate [78-44-4]. DEFINITION Carisoprodol contains NLT 98.0% and NMT 102.0% of C12H24N2O4, calculated on the dried basis.
260.33
USP 32
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 9194 LOSS ON DRYING 731: Dry it in vacuum at 60 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carisoprodol RS USP Meprobamate RS CU
Official Monographs / Carisoprodol 87

= nominal concentration of carisoprodol in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.05 M, pH 6.9 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions) containing 5 units of -amylase per mL; 900 mL [NOTEUse only freshly prepared solutions containing amylase; and equilibrate the Dissolution Medium at 37 for NMT one h before beginning the Dissolution test.] Apparatus 2: 75 rpm Time: 60 min Determine the amount of C12H24N2O4 dissolved employing the following method: Mobile phase, System suitability solution, and System suitability: Proceed as directed in the Assay. Standard solution: 0.4 mg/mL of USP Carisoprodol RS in Medium [NOTEA volume of acetonitrile not exceeding 2% of the final total volume of solution may be used to aid in dissolving the carisoprodol.] Sample solution: Sample per Dissolution 711. Filter before analyzing. Chromatographic system: Proceed as directed under Assay, except use the following injection size: Injection size: 150 L Analysis Samples: Standard solution and Sample solution Record the peak responses, and measure the heights for the major peaks. Calculate the amount of C12H24N2O4 dissolved. Tolerances: NLT 80% (Q) of the labeled amount of C12H24N2O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Carisoprodol RS
Carisoprodol Tablets
(Comment on this Monograph)id=m13380=Carisoprodol Tablets=Ca-Chl-Monos.pdf) DEFINITION Carisoprodol Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C12H24N2O4. IDENTIFICATION The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (2:3) Diluent: Methanol and 0.01 N sulfuric acid (3:2) System suitability solution: 2.4 mg/mL of 2-methyl-2propyl-1,3-propanediol and 3.4 mg/mL of carisoprodol in Mobile phase Standard solution: 3.5 mg/mL of USP Carisoprodol RS in Diluent Sample solution: Transfer an amount equivalent to 350 mg of carisoprodol from powdered Tablets (NLT 20 Tablets), to a 100-mL volumetric flask and add 50 mL of Diluent, place in an ultrasonic bath for 30 min, and shake by mechanical means for 60 min. Dilute with Diluent to volume. Pass a portion of this solution through a membrane filter of 0.5m or finer porosity, and use the filtrate as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: 3.9-mm 30-cm; packing L1 Temperature: 30 1 for column and detector Flow rate: 2 mL/min Injection size: 35 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for 2-methyl-2propyl-1,3-propanediol and carisoprodol are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the 2-methyl-2-propyl-1,3propanediol and carisoprodol peaks, System suitability solution Relative standard deviation: NMT 2.0% for three replicate injections of the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H24N2O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carisoprodol RS in the Standard solution (mg/mL)
Carisoprodol and Aspirin Tablets

(Comment on this Monograph)id=m13383=Carisoprodol and Aspirin Tablets=Ca-Chl-Monos.pdf) DEFINITION Carisoprodol and Aspirin Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of carisoprodol (C12H24N2O4) and aspirin (C9H8O4). IDENTIFICATION A. The retention times of the aspirin peak and the carisoprodol peak of the Sample solution correspond to those from Standard solution A, as obtained in Assay for Aspirin and carisoprodol. ASSAY ASPIRIN AND CARISOPRODOL Mobile phase: Methanol and 0.174 M acetic acid (16:9) Diluent: Acetonitrile, glacial acetic acid, and water (40:1:59) Standard solution A: To 80 mg of USP Aspirin RS and 80J mg of USP Carisoprodol RS, in a 25-mL volumetric flask, add 15 mL of Diluent, swirl for 5 min, and sonicate for 2530 s (J being the ratio of the labeled amount, in mg, of carisoprodol to that of aspirin). Dilute with Diluent to volume. Standard solution B: 16 g/mL of USP Salicylic Acid RS in Diluent
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USP 32
Mode: LC Detector: Refractive index Column: 3.9-mm 30-cm; packing L1 Temperature: 30 1 for column and detector Flow rate: 2 mL/min Injection size: 300 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for aspirin and carisoprodol are 0.4 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between the aspirin and salicylic acid peaks, and between the carisoprodol and salicylic acid peaks, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 and C12H24N2O4 dissolved: Result = (rU/rS) (CS/CU) 100 = peak response for aspirin or carisoprodol from the Sample solution = peak response for aspirin or carisoprodol from rS the Standard solution A = concentration of USP Aspirin RS or USP CS Carisoprodol RS in the Standard solution A (mg/mL) = nominal concentration of aspirin or carisoprodol CU in the Sample solution (mg/mL) Tolerances: NLT 75% (Q) of the labeled amounts of C9H8O4 and C12H24N2O4 are dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to aspirin and to carisoprodol rU IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase, Diluent, Standard solution A, Standard solution B, System suitability solution, and Sample solution: Proceed as directed in Assay. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 313 nm Column: 4.6-mm 25-cm; packing L7 Temperature: 30 1 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: Standard solution A, Standard solution B and System suitability solution [NOTEThe relative retention times for aspirin, salicylic acid, and carisoprodol are about 0.6, 0.7, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.2 between the solvent and aspirin peaks and NLT 1.5 between the aspirin and salicylic acid peaks from the System suitability solution Relative standard deviation: NMT 2.0% for Standard solution A; NMT 5.0% for Standard solution B Analysis Samples: Standard solution A, Standard solution B, and Sample solution
System suitability solution: 0.5 mg/mL of salicylic acid in Standard solution A Sample solution: To an equivalent to 325 mg of aspirin from powdered Tablets (NLT 20), in a 100-mL volumetric flask, add 50 mL of Diluent, swirl for 5 min, sonicate for 2530 s, shake by mechanical means for 30 min, dilute with Diluent to volume, and mix. Pass a portion of this solution through a membrane filter of 0.5m or finer porosity, and use the filtrate. [NOTEUse within 8 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: 4.6-mm 25-cm; packing L7 Temperature: 30 1 column and refractive index detector Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: Standard solution A, Standard solution B, and System suitability solution [NOTEThe relative retention times for aspirin, salicylic acid, and carisoprodol are about 0.6, 0.7, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.2 between the solvent and aspirin peaks and NLT 1.5 between the aspirin and salicylic acid peaks from the System suitability solution Relative standard deviation: NMT 2.0% for Standard solution A; NMT 5.0% for Standard solution B Analysis Samples: Standard solution A and Sample solution Analysis: Calculate the percentage of C9H8O4 and C12H24N2O4 in the portion of Tablets: Result = (rU/rS) (CS/CU) 100 = peak response for aspirin or carisoprodol from the Sample solution = peak response for aspirin or carisoprodol from rS the Standard solution A = concentration of USP Aspirin RS or USP CS Carisoprodol RS in the Standard solution A (mg/mL) = nominal concentration of aspirin or carisoprodol CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 45 min Mobile phase: Methanol and glacial acetic acid solution (1 in 50) (51:49) Standard solution: To 90 mg of USP Aspirin RS and 90J mg of USP Carisoprodol RS in a 250-mL volumetric flask, add 5 mL of acetonitrile, previously passed through a membrane filter of 0.5-m or finer porosity, swirl to dissolve, and dilute with water to volume (J being the ratio of the labeled amount, in mg, of carisoprodol to that of aspirin). System suitability solution: 0.36 mg/mL of salicylic acid in Standard solution Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) rU
USP 32
Calculate the percentage of free salicylic acid in the Tablets: Result = (rU/rS) (CS/CU) 100 = peak response for salicylic acid from the Sample solution = peak response for salicylic acid from the rS Standard solution B = concentration of USP Salicylic Acid RS in the CS Standard solution B (g/mL) = nominal concentration of aspirin in the portion CU of Tablets taken in the Sample solution (g/mL) Acceptance criteria: NMT 3.0% is found rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Carisoprodol RS USP Salicylic Acid RS
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Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: Standard solution A, Standard solution B, and System suitability solution [NOTEThe relative retention times for aspirin, salicylic acid, and carisoprodol are about 0.6, 0.7, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.2 between the solvent and aspirin peaks and NLT 1.5 between the aspirin and salicylic acid peaks from the System suitability solution Relative standard deviation: NMT 2.0% for Standard solution A; NMT 5.0% for Standard solution B Analysis Samples: Standard solution A and Sample solution Calculate the percentage of C9H8O4 and C12H24N2O4 in the portion of Tablets: Result = (rU/rS) (CS/CU) 100 = peak response for aspirin or carisoprodol from the Sample solution rS = peak response for aspirin or carisoprodol from the Standard solution A CS = concentration of USP Aspirin RS or US Carisoprodol RS in the Standard solution A (mg/mL) CU = nominal concentration of aspirin or carisoprodol in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amounts of C12H24N2O4 and C9H8O4 CODEINE PHOSPHATE Mobile phase: Dissolve 2.2 g of docusate sodium in 600 mL of methanol. Dissolve 0.8 g of ammonium nitrate in 400 mL of water. Mix these two solutions and adjust with glacial acetic acid to a pH of 3.3 0.05. Diluent: Methanol and 0.01 N sulfuric acid (1:1) System suitability solution: 0.16 mg/mL of USP Codeine Phosphate RS and 0.12 mg/mL of USP Codeine N-Oxide RS in Diluent Standard solution: 0.16 mg/mL of USP Codeine Phosphate RS and 0.16J mg/mL of USP Aspirin RS in Diluent, with the aid of swirling for 5 min and sonication for 2530 s (J being the ratio of the labeled amount, in mg, of aspirin to that of codeine phosphate) Sample solution: To an amount equivalent to 16 mg of codeine phosphate from powdered Tablets (NLT 20), in a 100-mL volumetric flask, add 50 mL of Diluent, sonicate for 30 min, shake by mechanical means for 30 min, and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for codeine N-Oxide and codeine phosphate are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.2 between the codeine phosphate and codeine N-Oxide peaks in the System suitability solution Relative standard deviation: NMT 2.0% from the Standard solution rU
Carisoprodol, Aspirin, and Codeine Phosphate Tablets

(Comment on this Monograph)id=m13385=Carisoprodol, Aspirin, and Codeine Phosphate Tablets=Ca-Chl-Monos.pdf) DEFINITION Carisoprodol, Aspirin, and Codeine Phosphate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of carisoprodol (C12H24N2O4), aspirin (C9H8O4), and codeine phosphate (C18H21NO3 H3PO4 1/2H2O). IDENTIFICATION A. The retention times of the aspirin, carisoprodol, and codeine phosphate peaks from the Sample solutions correspond to those of the Standard solutions obtained as directed in the Assay for Aspirin and Carisoprodol and the Assay for Codeine Phosphate ASSAY [NOTEBoth Aspirin and Carisoprodol and Codeine Phosphate must be completed for this test.] ASPIRIN AND CARISOPRODOL Mobile phase: Methanol and 0.174 M acetic acid (16:9) Diluent: Acetonitrile, glacial acetic acid, and water (40:1:59) Standard solution A: To 80 mg of USP Aspirin RS and 80J mg of USP Carisoprodol RS, in a 25-mL volumetric flask, add 15 mL of Diluent, swirl for 5 min, and sonicate for 2530 s. Dilute with Diluent to volume (J being the ratio of the labeled amount, in mg, of carisoprodol to that of aspirin). Standard solution B: 16 g/mL of USP Salicylic Acid RS in Diluent System suitability solution: 0.5 mg/mL of salicylic acid in Standard solution A Sample solution: To an amount equivalent to 325 mg of aspirin from powdered Tablets (NLT 20), in a 100-mL volumetric flask, add 50 mL of Diluent, swirl for 5 min, sonicate for 2530 s, shake by mechanical means for 30 min, dilute with Diluent to volume, and mix. Pass a portion of this solution through a membrane filter of 0.5m or finer porosity, and use the filtrate. [NOTEUse within 8 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: 4.6-mm 25-cm; packing L7 Temperature: 30 1 column and refractive index detector
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Carisoprodol / Official Monographs
USP 32
DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 45 min Procedure 1: Determination of dissolved aspirin and carisoprodol Mobile phase: Methanol and glacial acetic acid solution (1 in 50) (51:49) Standard solution: To 90 mg of USP Aspirin RS and 90J mg of USP Carisoprodol RS in a 250-mL volumetric flask, add 5 mL of acetonitrile, previously passed through a membrane filter of 0.5-m or finer porosity, swirl to dissolve, and dilute with water to volume (J being the ratio of the labeled amount, in mg, of carisoprodol to that of aspirin). System suitability solution: 0.36 mg/mL of salicylic acid in Standard solution Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: 3.9-mm 30-cm; packing L1 Temperature: 30 1 for column and detector Flow rate: 2 mL/min Injection size: 300 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for aspirin and carisoprodol are 0.4 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between the aspirin and salicylic acid peaks, and between the carisoprodol and salicylic acid peaks, in the System suitability solution Relative standard deviation: NMT 2.0% of the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H8O4 and C12H24N2O4 dissolved: Result = (rU/rS) (CS/CU) 100 = peak response for aspirin or carisoprodol from the Sample solution = peak response for aspirin or carisoprodol from rS the Standard solution A = concentration of USP Aspirin RS or USP CS Carisoprodol RS in the Standard solution A (mg/mL) = nominal concentration of aspirin or carisoprodol CU in the Sample solution (mg/mL) Tolerances: NLT 75% (Q) of the labeled amounts of C9H8O4 and C12H24N2O4 are dissolved. Procedure 2: Determination of dissolved codeine phosphate Mobile phase: Dissolve 2.2 g of docusate sodium and 0.8 g of ammonium nitrate in 550 mL of water, and pass through a membrane filter of 0.5-m or finer porosity. Add 450 mL of similarly filtered acetonitrile to the filtrate. Standard solution: 18 g/mL of USP Codeine Phosphate RS in water rU
Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H21NO3 H3PO4 1/2H2O in the portion of Tablets: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight of anhydrous codeine Mr2 phosphate, 397.37 Acceptance criteria: 90.0%110.0% of the labeled amount of C18H21NO3 H3PO4 1/2H2O rU rS CS IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE SALICYLIC ACID Mobile phase, Diluent, Standard solution A, Standard solution B, System suitability solution, and Sample solution: Proceed as directed in the Assay for Aspirin and Carisoprodol. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 313 nm Column: 4.6-mm 25-cm; packing L7 Temperature: 30 1 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: Standard solution A, Standard solution B and System suitability solution [NOTEThe relative retention times for aspirin, salicylic acid, and carisoprodol are about 0.6, 0.7, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.2 between the solvent and aspirin peaks and NLT 1.5 between the aspirin and salicylic acid peaks from the System suitability solution Relative standard deviation: NMT 2.0% for Standard solution A; NMT 5.0% for Standard solution B Analysis Samples: Standard solution A, Standard solution B, and Sample solution Calculate the percentage of free salicylic acid in the Tablets: Result = (rU/rS) (CS/CU) 100 = peak response for salicylic acid from the Sample solution rS = peak response for salicylic acid from the Standard solution B = concentration of USP Salicylic Acid RS in the CS Standard solution B (g/mL) = nominal concentration of aspirin in the portion of CU Tablets taken in the Sample solution (g/mL) Acceptance criteria: NMT 3.0% is found PERFORMANCE TESTS [NOTEBoth Procedure 1 and Procedure 2 must be completed for this test.] rU
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Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H21NO3 H3PO4 1/2H2O dissolved: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of codeine phosphate Mr1 hemihydrate, 406.37 = molecular weight of anhydrous codeine Mr2 phosphate, 397.37 Tolerances: NLT 75% (Q) of the labeled amounts of C18H21NO3 H3PO4 1/2H2O are dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to aspirin, carisoprodol, and codeine phosphate rU rS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Aspirin RS USP Carisoprodol RS USP Codeine N-Oxide RS USP Codeine Phosphate RS USP Salicylic Acid RS
Official Monographs / Carprofen 91

ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, glacial acetic acid, and water (40:25:0.2:35) System suitability stock solution: 16 g/mL of USP Carprofen Related Compound A RS in Mobile phase. Sonicate if necessary. [NOTEUse low-actinic glassware.] System suitability solution: Mix 10 mL of System suitability stock solution and 10 mL of the Standard solution and dilute with Mobile phase to 100 mL. [NOTEUse low-actinic glassware] Standard solution: 160 g/mL of USP Carprofen RS in Mobile phase. Sonicate if necessary. [NOTEUse low-actinic glassware.] Sample solution: 160 g/mL of Carprofen in Mobile phase [NOTEUse low-actinic glassware] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 239 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 2.0 between carprofen and carprofen related compound A Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H12ClNO2 in the portion of Carprofen taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carprofen RS in the Standard solution (g/mL) = concentration of carprofen in the Sample solution CU (g/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1: LIMIT OF ACETONE AND METHYLENE CHLORIDE Standard solution: 0.5 mg/mL of acetone and 0.06 mg/mL of methylene chloride in N,N-dimethylacetamide Sample solution: 100 mg/mL of Carprofen in N,Ndimethylacetamide Chromatographic system Mode: GC Detector: Flame ionization Column: 0.53-mm 30-m capillary column coated with 3.0-m G43 stationary phase rU rS CS
Carprofen
(Comment on this Monograph)id=m13600=Carprofen=Ca-ChlMonos.pdf)
273.71 C15H12ClNO2 9H-Carbazole-2-acetic acid, 6-chloro--methyl-, ()-; ()-6-Chloro--methylcarbazole-2-acetic acid [53716-49-7]. DEFINITION Carprofen contains NLT 98.0% and NMT 102.0% of C15H12ClNO2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
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Carrier gas: Nitrogen Temperature (See the temperature table below.)
Time (min) Temperature () 80 80 190 190 210 220 Impurity Name Carprofen related compound A (carbazole) 2-[1,1-Dimethoxy-2hydroxypropyl]-6chlorocarbazole 2-[2Chloropropionyl]-6chloro-9acetylcarbazole Any individual unknown related compound
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Acceptance criteria Individual impurities (See Impurity Table 1.) Total impurities: NMT 1.0%
Impurity Table 1 Relative Retention Time 0.9 Relative Response Factor Acceptance Criteria NMT (%) 0.5
Column
0 4 7.66 10.66
Injection port Detector
1.3
0.5
Flow rate: 4.9 mL/min Injection size: 1 L Injection type: Split flow ratio, 10:1 System suitability Sample: Standard solution [NOTEAcetone elutes before methylene chloride.] Suitability requirements Resolution: NLT 1.5 between acetone and methylene chloride Relative standard deviation: NMT 10.0% for the acetone peak responses Analysis Samples: Standard solution and Sample solution Calculate the percentage of each residual solvent in the portion of Carprofen taken: Result = (rU/rS) (CS/CU) 100 = peak response of the individual residual solvent in the Sample solution = peak response of the individual residual solvent rS in the Standard solution = concentration of individual residual solvent in CS the Standard solution (g/mL) = concentration of carprofen in the Sample CU solution (g/mL) Acceptance criteria Acetone: NMT 5000 ppm Methylene chloride: NMT 600 ppm Procedure 2 Mobile phase, Sample solution, System suitability solution, System suitability, and Chromatographic system: Proceed as directed in the Assay. Analysis Sample: Sample solution Calculate the percentage of each related compound in the portion of Carprofen taken: rU Result = (rU/rT) 100 rU rT = peak response of each individual peak (other than the major peak of carprofen) = sum of the peak responses
3.3
0.5
0.1
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at 25, excursions permitted between 15 and 30. LABELING: Label it to indicate that it is intended for veterinary use only. USP REFERENCE STANDARDS 11 USP Carprofen RS USP Carprofen Related Compound A RS
Carprofen Tablets
(Comment on this Monograph)id=m13610=Carprofen Tablets=Ca-Chl-Monos.pdf) DEFINITION Carprofen Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of carprofen (C15H12ClNO2). IDENTIFICATION A. INFRARED ABSORPTION 197K Standard: Mix 2 mg of USP Carprofen RS with 200 mg of potassium bromide, and grind thoroughly for 1015 min. Compress the mixture into a clear pellet. Record the IR spectrum of the pellet immediately after preparation. Sample: Transfer an amount equivalent to 100 mg of carprofen from powdered Tablets (NLT 4 Tablets), to a 125-
USP 32
mL separatory funnel. Add 30 mL of water and 3 drops of hydrochloric acid, and shake for 5 min. Add 30 mL of methylene chloride, and shake for another 5 min. Allow the phases to separate. Carefully drain and collect the lower methylene chloride layer through anhydrous sodium sulfate that is placed on a cotton pledget into a suitable container. Evaporate the methylene chloride on a steam bath with the aid of a stream of nitrogen to dryness. Dry the residue in vacuum at 60 for about 30 min. Mix 2 mg of the dried residue with 200 mg of potassium bromide, and grind thoroughly for 1015 min. Compress the mixture into a clear pellet. Record the IR spectrum of the carprofen sample pellet immediately after preparation. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, phosphoric acid, and water (500:1:500) Standard solution: 0.05 mg/mL of USP Carprofen RS in Mobile phase [NOTEUse low-actinic glassware.] Sample solution: Weigh 20 Tablets, and calculate the average Tablet weight. Grind the Tablets into a uniform powder. Transfer three weighed portions of the powder, each equivalent to the weight of one Tablet, into three volumetric flasks of a suitable calibrated volume such that an interim concentration of 0.5 mg/mL of Mobile phase can be prepared. To each flask, add Mobile phase to 80% of the calibrated volume, sonicate for 10 min, then stir for 10 min. Sonicate again for 10 min, and stir for another 10 min. Cool to room temperature, dilute with Mobile phase to volume to obtain an interim concentration of 0.5 mg of carprofen/mL, and mix. Quantitatively transfer 5.0 mL of the solution to a 50.0-mL volumetric flask, and dilute with Mobile phase to volume. Pass the solution through a PVDF filter having a 0.45-m or finer porosity, discarding the first 5 mL of the filtrate. The final concentration is 0.05 mg/mL of carprofen. [NOTEUse low-actinic glassware.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 20 L [NOTEWash the column after each series of analyses with a mixture of acetonitrile and water (20:80) for 30 min; gradually change the composition of acetonitrile and water to 80:20 over 10 min; continue to wash at 80:20 for 30 min; gradually change the composition to 50:50 over 10 min; and continue to wash at 50:50 for another 30 min.] System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 4000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% [NOTEInject the Standard solution in duplicate after every 12 injections or fewer of any other solution. The ratio of the average area of the duplicate injections to that obtained from the initial five replicate injections is 0.951.05.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of the labeled content of C15H12ClNO2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS Cs
Official Monographs / Carprofen 93

= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carprofen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution Cu (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amount of C15H12ClNO2 PERFORMANCE TESTS DISSOLUTION 711 [NOTEUse low-actinic volumetric flasks, dissolution vessels, and evaporation covers.] Medium: 0.05 M phosphate buffer, pH 7.5 (prepared by dissolving 6.8 g of monobasic potassium phosphate in 600 mL of water, mixing, adding 18 mL of 2 N sodium hydroxide, mixing, diluting with water to 1000 mL, and adjusting with 0.2 N sodium hydroxide or 0.2 N hydrochloric acid to a pH of 7.50 0.05); 900 mL Apparatus 2: 50 rpm Time: 30 min Determine the amount of C15H12ClNO2 dissolved by employing the following method. Standard solution: For Tablets labeled to contain 25 mg: 0.028 mg/mL of USP Carprofen RS in methanol and Medium (1:89) For Tablets labeled to contain 75 mg: 0.083 mg/mL of USP Carprofen RS in methanol and Medium (3:87) For Tablets labeled to contain 100 mg: 0.111 mg/mL of USP Carprofen RS in methanol and Medium (2:43) Sample solutions: Sample per 711 Dissolution. Dilute with Medium to a concentration that is similar to the Standard solution. Pass a portion of the solution under test through a suitable 0.45-m filter. System suitability solution: Determine the absorbance of the Standard solution, as directed for Analysis, five times: the relative standard deviation is NMT 2.0%. Analysis Samples: Standard solution, Sample solution, and Blank Determine the amount of C15H12ClNO2 dissolved by measuring the absorbance of the Sample solution in comparison with the appropriate Standard solution at the wavelength of maximum absorbance at 300 nm, using a 0.5-cm cell for Tablets labeled to contain 25 mg, a 0.2-cm cell for Tablets labeled to contain 75 mg, and a 0.1-cm cell for Tablets labeled to contain 100 mg. Use Medium as the blank. Calculate the percentage of C15H12ClNO2 dissolved: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Carprofen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Tolerances: NLT 80% (Q) of the labeled amount of C15H12ClNO2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity Procedure for content uniformity [NOTEUse low-actinic glassware.] Mobile phase, Standard solution, System suitability, and Chromatographic system: Prepare as directed in the Assay. Sample solution: Transfer 10 Tablets individually to 10 separate volumetric flasks of a suitable calibrated volume such that an interim concentration of 0.5 mg/mL of Mobile phase can be prepared. To each flask, add Mobile phase to 80% of the calibrated volume, sonicate for 10 min, then AU AS CS
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Carprofen / Official Monographs

stir for 10 min. Sonicate again for 10 min, and stir for another 10 min or until the Tablets are completely disintegrated. Cool to room temperature, dilute with Mobile phase to volume to obtain an interim concentration of 0.5 mg of carprofen/mL, and mix. Quantitatively transfer 5.0 mL of the individual solutions to 10 separate 50.0-mL volumetric flasks, dilute with Mobile phase to volume, and mix. Pass the solution through a polyvinylidenefluoride (PVDF) filter having a 0.45-m or finer porosity, discarding the first 5 mL of the filtrate. The final concentration is about 0.05 mg of carprofen/mL. Analysis: Proceed as directed for Analysis in the Assay. Calculate the percentage of the labeled content of C15H12ClNO2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak area response from the Sample solution = peak area response rom the Standard solution = concentration of USP Carprofen RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL)
USP 32
Column efficiency: NLT 4000 theoretical plates, Standard solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution [NOTEThe carprofen peak should be defined and integratable from injections of the System suitability solution.] [NOTEAfter every six injections of any solution, inject a Standard solution in duplicate. The ratio of the average response of the duplicate injections to that obtained from the initial five replicate injections is 0.951.05.] Analysis Samples: Standard solution, Sample solution and Blank solution Calculate the percentage of carprofen-related compounds in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak area of any peak other than carprofen from the Sample solution = peak area of carprofen from the Standard rS solution = concentration of carprofen in the Standard CS solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria Individual impurities: NMT 0.5% Total impurities: NMT 2.0% [NOTEDisregard any peak also observed in the Blank solution.] rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carprofen RS
IMPURITIES Organic Impurities PROCEDURE Mobile phase: Proceed as directed in the Assay. [NOTEUse low-actinic glassware.] Standard solution: 0.05 g/mL of USP Carprofen RS in Mobile phase System suitability solution: 0.005 g/mL of Carprofen, from Standard solution in Mobile phase Sample solution: Use the Sample solution, prepared as directed in the Assay. Blank solution: Transfer an amount of Tablet base equivalent to the weight of 1 Tablet to a volumetric flask of the same calibrated volume as that used to prepare the Sample solution. To each flask, add Mobile phase to 80% of the calibrated volume. Sonicate for 10 min, then stir for 10 min. Sonicate again for 10 min, and stir for another 10 min. Cool to room temperature, and dilute with Mobile phase to volume. Quantitatively transfer 5.0 mL of the solution to a 50.0-mL volumetric flask, dilute with Mobile phase to volume, and mix. Pass the solution through a PVDF filter having a 0.45-m or finer porosity, discarding the first 5 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 15-cm; 5-m packing L7 Flow rate: 1 mL/min Injection size: 50 L [NOTEWash the column after each series of analyses with a mixture of acetonitrile and water (20:80) for 30 min; gradually change the composition of acetonitrile and water to 80:20 over 10 min; continue to wash at 80:20 for 30 min; gradually change the composition to 50:50 over 10 min; and continue to wash at 50:50 for another 30 min.] System suitability Samples: Standard solution, Sample solution and System suitability solution Suitability requirements Resolution: NLT 2.0 between carprofen and the nearest impurity peak, Sample solution
Carteolol Hydrochloride
(Comment on this Monograph)id=m13660=Carteolol Hydrochloride=Ca-Chl-Monos.pdf)
328.83 C16H24N2O3 HCl 2(1H)-Quinolinone, 5-[3-[(1,1-dimethylethyl)amino]-2hydroxypropoxy]-3,4-dihydro-, monohydrochloride; 5-[3-(tert-Butylamino)-2-hydroxypropoxy]-3,4-dihydrocarbostyril monohydrochloride [51781-21-6]. DEFINITION Carteolol Hydrochloride contains NLT 98.0% and NMT 101.5% of C16H24N2O3 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Solution: 10 g/mL C. CHLORIDE AND SULFATE, Chloride 191: solution responds to the tests.
A 20 mg/mL
USP 32
ASSAY PROCEDURE Solution A: 0.67 mg/mL of dibasic sodium phosphate, adjust with 1 M phosphoric acid to a pH of 6.0 0.05 before bringing to volume Mobile phase: Acetonitrile and Solution A (1:3) [NOTEIncreasing the proportion of pH 6.0 buffer increases resolution.] Diluent: Methanol and Solution A (1:1) Standard stock solution: 1 mg/mL of USP Carteolol Hydrochloride RS Standard solution: Transfer 10.0 mg/mL of Standard stock solution to a 100mL volumetric flask containing 5 mL acetonitrile, and dilute with water to volume. System suitability stock solution: Dissolve 50 mg of pacetotoluidide in 50 mL of acetonitrile and dilute with water to 100 mL. System suitability solution: Combine 10.0 mL of System suitability stock solution and 10.0 mL of Standard stock solution. Dilute with water to 100 mL (contains 0.05 mg/mL of p-acetotoluidide and 0.1 mg/mL of USP Carteolol Hydrochloride RS). Sample stock solution: 1 mg/mL of Carteolol Hydrochloride Sample solution: Transfer 10.0 mL of Sample stock solution to a 100mL volumetric flask containing 5 mL of acetonitrile, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 252 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for carteolol and pacetotoluidide are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 between the carteolol peak and pacetotoluidide peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H24N2O3 HCl in the portion of Carteolol Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP carteolol hydrochloride RS in the Standard solution (mg/mL) CU = concentration of carteolol hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 98.0%101.5% rU rS CS SPECIFIC TESTS PH 791: 5.06.0, in a solution (1 in 100) LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 0.5% of its weight. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% ARSENIC, Method II 211: NMT 3 ppm HEAVY METALS, Method I 231: NMT 20 ppm Organic Impurities PROCEDURE Standard solution A: 0.5 mg/mL of USP Carteolol Hydrochloride RS in methanol
Official Monographs / Carteolol 95

Standard solution B: Dilute 5.0 mL of Standard solution A to 50 mL with methanol. Standard solution C: Dilute 5.0 mL of Standard solution B to 10 mL with methanol. Sample solution: Transfer 250 mg of Carteolol Hydrochloride to a 10mL volumetric flask, dissolve in methanol, using heat or sonication if necessary to achieve dissolution. Dilute with methanol to volume. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform, methanol, and ammonium hydroxide (50:20:1) Analysis Samples: Standard solutions A, B, and C; and Sample solution Proceed as directed in the chapter. Allow the spots to dry. Line a chromatographic chamber with filter paper, and saturate the paper with the Developing solvent system. Place the plate in the chamber, and develop the chromatogram until the solvent front has moved threefourths of the length of the plate. Remove the plate from the chamber, and allow to air-dry. Examine the plate under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from Standard solution A. Compare the sizes and intensities of any spots other than the principal spot from the Sample solution with those of the principal spots from the Standard solutions: no spot exceeds in size or intensity the principal spot from Standard solution B (0.2%), and the sum of all the impurity spots is NMT 0.5%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Carteolol Hydrochloride RS
Carteolol Hydrochloride Ophthalmic Solution

(Comment on this Monograph)id=m13665=Carteolol Hydrochloride Ophthalmic Solution=Ca-Chl-Monos.pdf) DEFINITION Carteolol Hydrochloride Ophthalmic Solution is a sterile, aqueous, isotonic solution of Carteolol Hydrochloride. It contains a suitable antimicrobial preservative. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C16H24N2O3 HCl. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 1 mg/mL of USP Carteolol Hydrochloride RS Sample solution: 1 mg/mL of carteolol hydrochloride from a volume of Ophthalmic Solution, diluted if necessary Application volume: 10 L Developing solvent system: Chloroform, methanol, and ammonium hydroxide (50:20:1) Analysis Samples: Standard solution and Sample solution Allow the spots to dry. Line a chromatographic chamber with filter paper, and saturate the paper with the Developing solvent system. Place the plate in the chamber,
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Carteolol / Official Monographs

CS
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= concentration of USP Carteolol Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of carteolol hydrochloride CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 6.08.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carteolol Hydrochloride RS
and develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, and allow to air-dry. Examine the plate under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that of the Standard solution. B. The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.67 mg/mL of dibasic sodium phosphate, adjust with 1 M phosphoric acid to a pH of 6.0 0.05, before bringing to volume Mobile phase: Acetonitrile and Solution A (1:3) [NOTE Increasing the proportion of pH 6.0 buffer increases resolution.] Diluent: Methanol and Solution A (1:1) Standard stock solution: 1 mg/mL of USP Carteolol Hydrochloride RS Standard solution: Transfer 10.0 mL of Standard stock solution, to a 100mL volumetric flask containing 5 mL of acetonitrile, and dilute with water to volume. System suitability stock solution: Dissolve 50 mg of pacetotoluidide in 50 mL of acetonitrile and dilute with water to 100 mL. System suitability solution: Combine 10.0 mL of System suitability stock solution, and 10.0 mL of Standard stock solution. Dilute with water to 100 mL. (Contains 0.05 mg/mL of p-acetotoluidide and 0.1 mg/mL of USP Carteolol Hydrochloride RS.) Sample solution: Transfer an amount of Ophthalmic Solution equivalent to 10 mg of carteolol hydrochloride to a 100mL volumetric flask and dilute with Diluent to volume. Pass a portion of this solution through a filter having a porosity of 0.5 m or finer, discarding the first 2 mL of the filtrate, and use the filtrate as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 252 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for carteolol and pacetotoluidide are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 between the carteolol peak and the pacetotoluidide peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H24N2O3 HCl in each mL of the Ophthalmic Solution taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
Carteolol Hydrochloride Tablets

(Comment on this Monograph)id=m13670=Carteolol Hydrochloride Tablets=Ca-Chl-Monos.pdf) DEFINITION Carteolol Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C16H24N2O3 HCl. IDENTIFICATION The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.67 mg/mL of dibasic sodium phosphate, adjust with 1 M phosphoric acid to a pH of 6.0 0.05 before bringing to volume Mobile phase: Acetonitrile and Solution A (1:3) [NOTEIncreasing the proportion of pH 6.0 buffer increases resolution.] Diluent: Methanol and Solution A (1:1) System suitability stock solution: Dissolve 50 mg of pacetotoluidide in 50 mL of acetonitrile and dilute with water to 100 mL. System suitability solution: Combine 10.0 mL of System suitability stock solution, and 10.0 mL of Standard stock solution. Dilute with water to 100 mL. (Contains 0.05 mg/mL of p-acetotoluidide and 0.1 mg/mL of USP Carteolol Hydrochloride RS.) Standard stock solution: 1 mg/mL of USP Carteolol Hydrochloride RS Standard solution: Transfer 10.0 mL of Standard stock solution to a 100mL volumetric flask containing 5 mL of acetonitrile, and dilute with water to volume Sample solution: Transfer an amount equivalent to 10 mg of carteolol hydrochloride from NLT 20 powdered Tablets to a 100mL flask. Add 50 mL of Diluent, and shake by mechanical means for 1 h. Add 5 mL of acetonitrile, and dilute with Diluent to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity, discarding the first 2 mL of filtrate, and use the clear filtrate as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 252 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 3 between the carteolol peak and the pacetotoluidide peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution [NOTEThe relative retention times for carteolol and pacetotoluidide are about 0.8 and 1.0, respectively.] Analysis Samples: Standard solution and Sample solution [NOTEUse peak areas where peak responses are indicated.] Calculate the percentage of C16H24N2O3 HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Carteolol Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of carteolol hydrochloride CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water, 900 mL Apparatus 2: 50 rpm Time: 30 min Solution A: 2.0 mg/mL of monobasic potassium phosphate Mobile phase: Acetonitrile and Solution A (2:3) Standard solution: 1.1L g/mL of USP Carteolol Hydrochloride RS, L being the labeled amount, in mg, of carteolol hydrochloride/Tablet Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to Standard solution. [NOTEPass a portion of the Sample solution through a filter having a porosity of 1 m or finer, discarding the first 2 mL of the filtrate.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 252 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 15 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H24N2O3 HCl dissolved: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Carteolol Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of carteolol hydrochloride CU in the Sample solution (g/mL) Acceptance criteria: NLT 80% (Q) of the labeled amount of C16H24N2O3 HCl rU rS CS rU rS CS
Official Monographs / Casanthranol 97

IMPURITIES Organic Impurities PROCEDURE: LIMIT OF DEHYDROCARTEOLOL HYDROCHLORIDE Solution A, Mobile phase, and Diluent: Proceed as directed in the Assay. Standard solution: 1 g/mL of USP Dehydrocarteolol Hydrochloride RS in Diluent Sample solution: Transfer an amount equivalent to 10 mg of carteolol hydrochloride from NLT 20 powdered Tablets to a 100mL volumetric flask, add 50 mL of Diluent, and shake by mechanical means for 1 h. Dilute with Diluent to volume. Pass a portion of this solution through a filter having a 0.5-m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Fluorometric detector, with excitation at 300 nm and a 418-nm emission filter Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 5.% Analysis Samples: Standard solution and Sample solution [NOTEUse peak areas where peak responses are indicated.] Calculate the percentage of dehydrocarteolol hydrochloride in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Dehydrocarteolol Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of carteolol hydrochloride CU in the Sample solution (mg/mL) Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Carteolol Hydrochloride RS USP Dehydrocarteolol Hydrochloride RS rU rS CS
Casanthranol
(Comment on this Monograph)id=m13700=Casanthranol=CaChl-Monos.pdf) DEFINITION Casanthranol is obtained from Cascara Sagrada. It contains in each 100 g NLT 20.0 g of total hydroxyanthracene derivatives calculated on the dried basis, calculated as cascaroside A. NLT 80% of the total hydroxyanthracene derivatives consists of cascarosides, calculated as cascaroside A. ASSAY TOTAL HYDROXYANTHRACENE DERIVATIVES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.6 for the ratio of the
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absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this Assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed under Reagents, Indicators, and SolutionsVolumetric Solutions.] Solution A: 1 g/mL of ferric chloride Sample stock solution: Transfer 500 mg of Casanthranol to a 100-mL volumetric flask. Add 30 mL of 70% alcohol, swirl to dissolve, and dilute with 70% alcohol to volume. Quickly filter through soft, rapid-flow filter paper, taking precautions to minimize loss by evaporation. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of 1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined aqueous phase with 30 mL of clear, freshly prepared water-saturated ethyl acetate, and transfer the water layer to another separatory funnel. Repeat the extraction with two additional 30-mL portions of the freshly prepared water-saturated ethyl acetate. Add 5 mL of water to the combined ethyl acetate extracts, shake, allow the phases to separate, discard the ethyl acetate extracts, and add 30 mL of the freshly prepared water-saturated ethyl acetate to the water wash. Shake, allow the phases to separate, and discard the ethyl acetate phase. Transfer the combined aqueous phases, with the aid of water, to a 50-mL volumetric flask, filtering through a small pledget of cotton, water-wet, and dilute with water to volume. Spectrometric conditions Mode: Visible Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Pipet 15 mL of Sample solution into a flask containing 2 mL of Solution A and 12 mL of hydrochloric acid. Attach a condenser arranged for refluxing, and heat for 3 h by keeping the flask immersed in boiling water or continuously exposed to steam heat. Cool, wash down the condenser, and transfer to a separatory funnel with the aid of 4 mL of 1 N sodium hydroxide and five 6-mL portions of water. Extract with 20 mL of methylene chloride, and transfer the lower layer to another separatory funnel. Repeat the extraction with three additional 20-mL portions of methylene chloride, wash the combined methylene chloride extracts with two 10-mL portions of water, shaking each time for 2 min, and discard the water washings. Transfer the washed methylene chloride extract to a 100-mL volumetric flask, dilute with methylene chloride to volume, and mix. Evaporate a 20.0-mL portion carefully on a water bath to dryness, and dissolve the residue in 10.0 mL of a 1in200 solution of magnesium acetate in methanol. Determine the absorbance and calculate the quantity, in mg, of cascarosides in the portion of Casanthranol taken: Result = AU F
absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this Assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed under Reagents, Indicators, and SolutionsVolumetric Solutions.] Solution A: 1 g/mL of ferric chloride Sample stock solution: Transfer 500 mg of Casanthranol to a 100-mL volumetric flask. Add 30 mL of 70% alcohol, swirl to dissolve, and dilute with 70% alcohol to volume. Quickly filter through soft, rapid-flow filter paper, taking precautions to minimize loss by evaporation. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of 1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, and discard the lower layer. Transfer the combined water layers, with the aid of water, to a 50-mL volumetric flask, filtering through a small pledget of cotton, water-wet, and dilute with water to volume. Spectrometric conditions Mode: Visible Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Pipet 10 mL of Sample solution into a flask containing 2 mL of Solution A and 12 mL of hydrochloric acid. Attach a condenser arranged for refluxing, and heat for 3 h by keeping the flask immersed in boiling water or continuously exposed to steam heat. Cool, wash down the condenser, and transfer to a separatory funnel with the aid of 4 mL of 1 N sodium hydroxide and five 6-mL portions of water. Extract with 20 mL of methylene chloride, and transfer the lower layer to another separatory funnel. Repeat the extraction with three additional 20-mL portions of methylene chloride, wash the combined methylene chloride extracts with two 10-mL portions of water, shaking each time for 2 min, and discard the water washings. Transfer the washed methylene chloride extract to a 100-mL volumetric flask, and dilute with methylene chloride to volume. Evaporate a 20.0-mL portion carefully on a water bath to dryness, and dissolve the residue in 10.0 mL of a 1in200 solution of magnesium acetate in methanol. Determine the absorbance and calculate the quantity, in mg, of total hydroxyanthracene derivatives in the portion of Casanthranol taken: Result = AU F = absorbance of the Sample solution AU F = monograph correction factor, 155 Acceptance criteria: NLT 20.0 g of total hydroxyanthracene derivatives/100 g calculated on the dried basis, calculated as cascaroside A. CASCAROSIDES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.7 for the ratio of the
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AU = absorbance of the Sample solution F = monograph correction factor, 103.5 mg Acceptance criteria: NLT 80% of the Total Hydroxyanthracene Derivatives IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 4.0% HEAVY METALS, Method II 231: NMT 25 ppm SPECIFIC TESTS LOSS ON DRYING 731: Dry it in vacuum at 80 for 16 h: it loses NMT 10.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at a temperature not exceeding 30.
Official Monographs / Cascara 99

1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined aqueous phase with 30 mL of clear, freshly prepared water-saturated ethyl acetate, and transfer the water layer to another separatory funnel. Repeat the extraction with two additional 30-mL portions of the freshly prepared water-saturated ethyl acetate. Add 5 mL of water to the combined ethyl acetate extracts, shake, allow the phases to separate, discard the ethyl acetate extracts, and add 30 mL of the freshly prepared water-saturated ethyl acetate to the water wash. Shake, allow the phases to separate, and discard the ethyl acetate phase. Transfer the combined aqueous phases, with the aid of water, to a 50-mL volumetric flask. Dilute with water to volume. Spectrometric system Mode: Visible Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Pipet 15 mL of Sample solution into a flask containing 2 mL of Solution A and 12 mL of hydrochloric acid. Attach a condenser arranged for refluxing, and heat for 3 h by keeping the flask immersed in boiling water or continuously exposed to steam heat. Cool, wash down the condenser, and transfer to a separatory funnel with the aid of 4 mL of 1 N sodium hydroxide and five 6-mL portions of water. Extract with 20 mL of methylene chloride, and transfer the lower layer to another separatory funnel. Repeat the extraction with three additional 20-mL portions of methylene chloride, wash the combined methylene chloride extracts with two 10-mL portions of water, shaking each time for 2 min, and discard the water washings. Transfer the washed methylene chloride extract to a 100-mL volumetric flask, and dilute with methylene chloride to volume. Evaporate a 20.0-mL portion carefully on a water bath to dryness, and dissolve the residue in 10.0 mL of a solution (1 in 200) of magnesium acetate in methanol Determine the absorbance and calculate the amount, in mg, of cascarosides in the portion of Cascara Sagrada taken: Result = F AU F = conversion factor, 103.5 = absorbance of the Sample solution AU Acceptance criteria: NLT 60% of the total hydroxyanthracene derivatives consists of cascarosides, calculated as cascaroside A. TOTAL HYDROXYANTHRACENE DERIVATIVES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.6 for the ratio of the absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this Assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed under Reagents, Indicators, and SolutionsVolumetric Solutions.]
Cascara Sagrada
(Comment on this Monograph)id=m13720=Cascara Sagrada=Ca-Chl-Monos.pdf) DEFINITION Cascara Sagrada is the dried bark of Rhamnus purshiana De Candolle (Fam. Rhamnaceae). It yields NLT 7.0% of total hydroxyanthracene derivatives, calculated as cascaroside A, and calculated on the dried basis. NLT 60% of the total hydroxyanthracene derivatives consists of cascarosides, calculated as cascaroside A. [NOTECollect Cascara Sagrada NLT 1 year before use.] IDENTIFICATION A. PROCEDURE Sample: 100 mg of powdered Cascara Sagrada Analysis: Add Sample to 10 mL of hot water, shake the mixture occasionally until it is cold, filter, dilute the filtrate with water to 10 mL, and add 10 mL of 6 N ammonium hydroxide. Acceptance criteria: An orange color is produced. B. It becomes red to reddish brown in color when treated with 6 N ammonium hydroxide. C. PROCEDURE Sample: 100 mg of powdered Cascara Sagrada Analysis: Macerate Sample with 1 mL of alcohol, add 10 mL of water, boil the mixture, then cool, filter, and shake the filtrate with 10 mL of ether: a greenish yellow ether solution separates. Shake 3 mL of this ether solution with 3 mL of 6 N ammonium hydroxide, and dilute the separated ammonia solution with 20 mL of water. Acceptance criteria: A distinct orange-pink color remains. ASSAY CASCAROSIDES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.7 for the ratio of the absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this Assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed under Reagents, Indicators, and SolutionsVolumetric Solutions.] Solution A: 1 g/mL of ferric chloride Sample stock solution: Add 1 g of Cascara Sagrada to 70 mL of boiling water, boil for several min, with stirring, allow to cool, and transfer with the aid of water to a 100-mL volumetric flask. Dilute with water to volume, and filter through suitable filter paper. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of
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an alkali; starch grains spheroidal, up to 8 m in diameter; calcium oxalate in monoclinic prisms or rosette aggregates from 6 to 20 m in diameter, occasionally up to 45 m in diameter. WATER DETERMINATION, Method IIIProcedure for Articles of Botanical Origin 921: Dry it at 105 for 5 h: it loses NMT 12.0% of its weight.
Solution A and Sample stock solution: Prepare as directed in the Assay for Cascarosides. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of 1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, and discard the lower layer. Transfer the combined water layers, with the aid of water, to a 50-mL volumetric flask, and dilute with water to volume. Spectrometric system Mode: Visible Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Proceed as directed for Analysis in the Assay for Cascarosides, except to evaporate a 15.0-mL portion of the methylene chloride solution instead of 20.0 mL. Calculate the quantity, in mg, of total hydroxyanthracene derivatives in the portion of Cascara Sagrada taken: Result = F AU F = conversion factor, 138 = absorbance of the Sample solution AU Acceptance criteria: NLT 7.0% of the total hydroxyanthracene derivatives, calculated as cascaroside A, and calculated on the dried basis. IMPURITIES Organic Impurities PROCEDURE: ARTICLES OF BOTANICAL ORIGIN, Foreign Organic Matter 561: NMT 4.0% SPECIFIC TESTS BOTANIC CHARACTERISTICS Cascara Sagrada: Usually in flattened or transversely curved pieces, occasionally in quills of variable length and from 1 to 5 mm in thickness. The outer surface is brown, purplish brown, or brownish red, longitudinally ridged, with or without grayish or whitish lichen patches, sometimes with numerous lenticels and occasionally with moss attached. The inner surface is longitudinally striate, light yellow, weak reddish brown, or moderate yellowish brown. The fracture is short with projections of phloem fiber bundles in the inner bark. Histology: It shows a yellowish brown, purple, or reddish brown cork of up to 10 or more rows of small cells; stone cells in yellowish, tangentially elongated groups of 2050 cells in the cortex, pericycle, and outer phloem regions; phloem rays 14 cells wide, 1525 cells deep, frequently diagonal or curved, forming converging groups; phloem fibers in small bundles, more or less surrounded by crystal fibers and located between the phloem rays; parenchyma with brown walls and containing starch grains and calcium oxalate crystals. Powdered Cascara Sagrada: Moderate yellowish brown to dusky yellowish orange. It shows numerous broken phloem fiber bundles with accompanying crystal fibers containing monoclinic prisms of calcium oxalate; stone cells more or less adherent, in small groups with thick, finely lamellated and porous walls; fragments of reddish brown to yellow cork; masses of parenchyma and phloem ray cells colored reddish brown to orange upon the addition of a solution of
Cascara Sagrada Extract

(Comment on this Monograph)id=m13730=Cascara Sagrada Extract=Ca-Chl-Monos.pdf) DEFINITION Cascara Sagrada Extract contains, in each 100 g, NLT 10.0 g and NMT 12.0 g of hydroxyanthracene derivatives, of which NLT 50% consists of cascarosides, both calculated as cascaroside A. Mix 900 g of Cascara Sagrada, in coarse powder, with 4000 mL of boiling water, and macerate the mixture for 3 h. Then transfer it to a percolator, allow it to drain, exhaust it by percolation, using boiling water as the menstruum, and collect 5000 mL of percolate. Evaporate the percolate to dryness, reduce the Extract to a fine powder, and, after assaying, add sufficient starch, dried at 100, or other inert, nontoxic diluents to make the product contain, in each 100 g, 11 g of hydroxyanthracene derivatives. Mix the powders, and pass the Extract through a number 60 sieve. ASSAY CASCAROSIDES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.7 for the ratio of the absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed for Reagents, Indicators, and SolutionsVolumetric Solutions.] Solution A: 1 g/mL of ferric chloride Sample stock solution: Transfer 1 g of Extract to a 100-mL volumetric flask. Add 60 mL of 70% alcohol, swirl or sonicate for 1520 min, several times, allow to stand overnight, sonicate or swirl for 1015 min, dilute with 70% alcohol to volume, mix, and filter through suitable filter paper. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of 1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined aqueous phase with 30 mL of clear, freshly prepared water-saturated ethyl acetate, and transfer the water layer to another separatory funnel. Repeat the extraction with two additional 30-mL portions of the freshly prepared water-saturated ethyl acetate. Add 5 mL of water to the combined ethyl acetate extracts, shake, allow the phases to separate, discard the ethyl acetate extracts, and add 30 mL of the freshly prepared water-saturated ethyl acetate to the water wash. Shake, allow the phases to separate, and discard the ethyl acetate phase. Transfer the combined aqueous phases, with
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the aid of water, to a 50-mL volumetric flask. Dilute with water to volume. Spectrometric conditions Mode: Visible Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Pipet 25 mL of Sample solution into a flask containing 2 mL of Solution A and 12 mL of hydrochloric acid. Attach a condenser arranged for refluxing, and heat for 3 h by keeping the flask immersed in boiling water or continuously exposed to steam heat. Cool, wash down the condenser, and transfer to a separatory funnel with the aid of 4 mL of 1 N sodium hydroxide and five 6-mL portions of water. Extract with 20 mL of methylene chloride, and transfer the lower layer to another separatory funnel. Repeat the extraction with three additional 20-mL portions of methylene chloride, wash the combined methylene chloride extracts with two 10-mL portions of water, shaking each time for 2 min, and discard the water washings. Transfer the washed methylene chloride extract to a 100-mL volumetric flask, dilute with methylene chloride to volume, and mix. Evaporate a 20.0-mL portion carefully on a water bath to dryness, and dissolve the residue in 10.0 mL of a 1in200 solution of magnesium acetate in methanol. Determine the absorbance and calculate the quantity, in mg, of Cascarosides in the portion of Extract taken: Result = F AU = correction factor, 62.06 = absorbance of the solution from the Sample solution Acceptance criteria: NLT 50% of Total Hydroxyanthracene Derivatives calculated as cascaroside A. TOTAL HYDROXYANTHRACENE DERIVATIVES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.6 for the ratio of the absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed for Reagents, Indicators, and SolutionsVolumetric Solutions..] Solution A and Sample stock solution: Prepare as directed in the Assay for Cascarosides. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of 1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, and discard the lower layer. Transfer the combined water layers, with the aid of water, to a 50-mL volumetric flask, and dilute with water to volume. F AU
Official Monographs / Cascara 101

Spectrometric conditions Mode: Visible Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Analysis: Pipet 10 mL of Sample solution into a flask containing 2 mL of Solution A and 12 mL of hydrochloric acid. Attach a condenser arranged for refluxing, and heat for 3 h by keeping the flask immersed in boiling water or continuously exposed to steam heat. Cool, wash down the condenser, and transfer to a separatory funnel with the aid of 4 mL of 1 N sodium hydroxide and five 6-mL portions of water. Extract with 20 mL of methylene chloride, and transfer the lower layer to another separatory funnel. Repeat the extraction with three additional 20-mL portions of methylene chloride, wash the combined methylene chloride extracts with two 10-mL portions of water, shaking each time for 2 min, and discard the water washings. Transfer the washed methylene chloride extract to a 100-mL volumetric flask, and dilute with methylene chloride to volume. Evaporate a 20.0-mL portion carefully on a water bath to dryness, and dissolve the residue in 10.0 mL of a 1in200 solution of magnesium acetate in methanol. Determine the absorbance and calculate the quantity, in mg, of Total Hydroxyanthracene Derivatives in the portion of Extract taken: Result = F AU = correction factor, 155.2 = absorbance of the solution from the Sample solution Acceptance criteria: 10.0 g12.0 g of hydroxyanthracene derivatives/100 g calculated as cascaroside A ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at a temperature not exceeding 30. F AU
Cascara Tablets
(Comment on this Monograph)id=m13740=Cascara Tablets=CaChl-Monos.pdf) DEFINITION Cascara Tablets are prepared from Cascara Sagrada Extract. They contain an amount of hydroxyanthracene derivatives, calculated as cascaroside A, NLT 9.35% and NMT 12.65% of the labeled amount of Cascara Sagrada Extract. NLT 50% of the hydroxyanthracene derivatives are cascarosides, calculated as cascaroside A. ASSAY CASCAROSIDES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.7 for the ratio of the
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F AU
USP 32
= monograph correction factor 103.5 = absorbance of the solution from the Sample solution Acceptance criteria: NLT 50% of the Total Hydroxyanthracene Derivatives, calculated as cascaroside A TOTAL HYDROXYANTHRACENE DERIVATIVES [NOTE 1Perform all extractions by shaking vigorously, and allow all phases to separate completely before transferring. Entrainment of aglycones into the aqueous phase, as indicated by a value of less than 2.6 for the ratio of the absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed under Reagents, Indicators, and Solutions, Volumetric Solutions.] Solution A and Sample stock solution: Prepare as directed in the Assay for Cascarosides. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of 1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, and discard the lower layer. Transfer the combined water layers, with the aid of water, to a 50-mL volumetric flask, and dilute with water to volume. Spectrometric conditions Mode: Spectrometry Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Prepare as directed under Analysis in the Assay for Cascarosides, except pipet 10 mL of Sample solution. Continue to and dilute with methylene chloride to volume. Evaporate a 15.0-mL portion carefully on a water bath to dryness, and dissolve the residue in 10.0 mL of a 1in200 solution of magnesium acetate in methanol. Determine the absorbance and calculate the quantity, in mg, of total hydroxyanthracene derivatives in the portion of Extract: Result = F AU = monograph correction factor 206.9 = absorbance of the solution from the Sample solution Acceptance criteria: Contains an amount of hydroxyanthracene derivatives NLT 9.35% and NMT 12.65% of the labeled amount of Cascara Sagrada Extract calculated as cascaroside A PERFORMANCE TESTS DISINTEGRATION 701: 60 min UNIFORMITY OF DOSAGE UNITS 905: F AU
absorbance of the final solution at 515 nm to that at 440 nm, may lead to false results.] [NOTE 2Throughout this assay, use 1 N sodium hydroxide that is prepared without added barium ions as directed for Reagents, Indicators, and Solutions, Volumetric Solutions.] Solution A: 1 g/mL of ferric chloride Sample stock solution: Transfer the equivalent to 1 g of Cascara Sagrada Extract from powdered Tablets (NLT 20), to a 100-mL volumetric flask. Add 60 mL of 70% alcohol, swirl or sonicate for 1520 min, several times, allow to stand overnight, sonicate or swirl for 1015 min, dilute with 70% alcohol to volume, mix, and filter through suitable filter paper. Sample solution: Pipet 10 mL of Sample stock solution into a separatory funnel containing 5 mL of water and 2 drops of 1 N hydrochloric acid. Extract with 40 mL of methylene chloride, and transfer the lower layer to a second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, discard the lower layer, and transfer the water layer to the first separatory funnel. Extract the combined water layers with 40 mL of methylene chloride, and transfer the lower layer to the second separatory funnel. Add 10 mL of water to the second separatory funnel, and shake. Allow to separate, and discard the lower layer. Transfer the water layer to the first separatory funnel. Extract the combined aqueous phase with 30 mL of clear, freshly prepared water-saturated ethyl acetate, and transfer the water layer to another separatory funnel. Repeat the extraction with two additional 30-mL portions of the freshly prepared water-saturated ethyl acetate. Add 5 mL of water to the combined ethyl acetate extracts, shake, allow the phases to separate, discard the ethyl acetate extracts, and add 30 mL of the freshly prepared watersaturated ethyl acetate to the water wash. Shake, allow the phases to separate, and discard the ethyl acetate phase. Transfer the combined aqueous phases, with the aid of water, to a 50-mL volumetric flask. Dilute with water to volume. Spectrometric conditions Mode: Spectrometry Analytical wavelength: 515 nm Cell: 1 cm Blank: Methanol Analysis Sample: Sample solution Pipet 20 mL of Sample solution into a flask containing 2 mL of Solution A and 12 mL of hydrochloric acid. Attach a condenser arranged for refluxing, and heat for 3 h by keeping the flask immersed in boiling water or continuously exposed to steam heat. Cool, wash down the condenser, and transfer to a separatory funnel with the aid of 4 mL of 1 N sodium hydroxide and five 6-mL portions of water. Extract with 20 mL of methylene chloride, and transfer the lower layer to another separatory funnel. Repeat the extraction with three additional 20-mL portions of methylene chloride, wash the combined methylene chloride extracts with two 10-mL portions of water, shaking each time for 2 min, and discard the water washings. Transfer the washed methylene chloride extract to a 100-mL volumetric flask, and dilute with methylene chloride to volume. Evaporate a 20.0-mL portion carefully on a water bath to dryness, and dissolve the residue in 10.0 mL of a 1in200 solution of magnesium acetate in methanol. Determine the absorbance and calculate the quantity, in mg, of cascarosides in the portion of Extract: Result = F AU
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers; if the Tablets are coated, well-closed containers may be used.
USP 32
Official Monographs / Castor 103
Cascara Sagrada Fluidextract

(Comment on this Monograph)id=m13750=Cascara Sagrada Fluidextract=Ca-Chl-Monos.pdf) DEFINITION Prepare Cascara Sagrada Fluidextract as follows. To 1000 g of coarsely ground Cascara Sagrada add 3000 mL of boiling water, and allow to macerate in a suitable percolator for 2 h. Allow the percolation to proceed at a moderate rate, gradually adding boiling water until the drug is practically exhausted of its active principles. Evaporate the percolate on a water bath or in a vacuum still to NMT 800 mL, cool, add 200 mL of alcohol and, if necessary, add sufficient water to make the product measure 1000 mL. OTHER COMPONENTS ALCOHOL DETERMINATION, Method I 611: C2H5OH 18.0%20.0% of
Castor Oil
(Comment on this Monograph)id=m13790=Castor Oil=Ca-ChlMonos.pdf) DEFINITION Castor Oil is the fixed oil obtained from the seed of Ricinus communis Linn (Fam. Euphorbiaceae). It contains no added e substances. IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231:
NMT 10 ppm
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and avoid exposure to direct sunlight and to excessive heat
Aromatic Cascara Fluidextract

(Comment on this Monograph)id=m13760=Aromatic Cascara Fluidextract=Ca-Chl-Monos.pdf) DEFINITION Prepare Aromatic Cascara Fluidextract as follows:
Cascara Sagrada, in very coarse powder Magnesium Oxide Suitable sweetening agent(s) Suitable essential oils(s) Suitable flavoring agent(s) Alcohol Purified Water, a sufficient quantity, to make 200 mL 1000 mL 1000 g 120 g
Mix the Cascara Sagrada with the Magnesium Oxide, moisten it uniformly with 2000 mL of boiling water, and set it aside in a shallow container for 48 h, stirring it occasionally. Pack it in a percolator, and percolate with boiling water until the drug is exhausted. Evaporate the percolate, at a temperature not exceeding 100, to 750 mL, and at once dissolve in it the flavoring agent(s). When the liquid has cooled, add the Alcohol, in which the sweetening agent(s) and oils have been dissolved, add sufficient water to make the Aromatic Fluidextract measure 1000 mL, and mix. OTHER COMPONENTS ALCOHOL DETERMINATION 611: Between 18%20% of C2H5OH, determined by the gasliquid chromatographic method, acetone being used as the internal standard ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and avoid exposure to direct sunlight and to excessive heat.
SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.9570.961 DISTINCTION FROM MOST OTHER FIXED OILS: It is only partly soluble in solvent hexane (distinction from most other fixed oils), but it yields a clear liquid with an equal volume of alcohol (foreign fixed oils). FATS AND FIXED OILS, Free Fatty Acids 401: The free fatty acids in 10 g require NMT 3.5 mL of 0.10 N sodium hydroxide for neutralization. FATS AND FIXED OILS, Hydroxyl Value 401 Sample solution: Transfer 2 g to a glass-stoppered, 250-mL conical flask. Add 5.0 mL of a freshly prepared mixture of acetic anhydride and pyridine (1:3), and swirl to mix. Analysis: Connect the flask to a reflux condenser, and heat on a steam bath for 2 h. Add 10 mL of water through the condenser, swirl to mix, heat on a steam bath for an additional 10 min, and allow to cool to room temperature. Add through the condenser 15 mL of normal butyl alcohol that previously has been neutralized to phenolphthalein, remove the condenser, and wash the tip of the condenser and the sides of the flask with an additional 10 mL of neutralized normal butyl alcohol. Add 1 mL of phenolphthalein TS, and titrate with 0.5 N alcoholic potassium hydroxide VS to a faint pink endpoint. Perform a blank determination on a 5.0-mL portion of the acetic anhydridepyridine mixture. To determine the amount of free acid in the Oil, weigh accurately 10 g into a 250-mL conical flask, add 10 mL of pyridine that previously has been neutralized to phenolphthalein, swirl to mix, add 1 mL of phenolphthalein TS, and titrate with 0.5 N alcoholic potassium hydroxide VS to a faint pink endpoint. Calculate the hydroxyl value in the portion of Oil taken: Result = 56.1N[A + (BW/D) C]/W = normality determined for the alcoholic potassium hydroxide solution A = volume of 0.5 N alcoholic potassium hydroxide consumed by the blank (mL) B = volume consumed in the free-acid titration (mL) W = weight of Oil taken (g) D = weight of Oil used in the free-acid titration (g) C = volume consumed in the sample titration (mL) Acceptance criteria: 160168 FATS AND FIXED OILS, Iodine Value 401: 8388 FATS AND FIXED OILS, Saponification Value 401: 176182 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid exposure to excessive heat. N
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Castor / Official Monographs
USP 32
Castor Oil Capsules

(Comment on this Monograph)id=m13800=Castor Oil Capsules=Ca-Chl-Monos.pdf) DEFINITION Castor Oil Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of castor oil, calculated from the tests for Weight variation and Specific gravity. IDENTIFICATION INFRARED ABSORPTION 197S Standard solution: A similar solution of Castor Oil Sample solution: 40 mg/mL of castor oil from Capsules in chloroform PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Castor Oil Emulsion

(Comment on this Monograph)id=m13810=Castor Oil Emulsion=Ca-Chl-Monos.pdf) DEFINITION Castor Oil Emulsion contains NLT 90.0% and NMT 120.0% of the labeled amount of Castor Oil. IDENTIFICATION PROCEDURE Analysis: Shake well, and place 10 mL in a 125-mL separator. Add 10 mL of 1 N hydrochloric acid and 20 mL of solvent hexane. Shake vigorously for 23 min, allow the layers to separate, discard the aqueous phase, and filter the upper layer through anhydrous sodium sulfate into a small beaker. Evaporate the solvent on a steam bath, and to the residue add 12 drops of sulfuric acid. Acceptance criteria: A red color indicates the presence of castor oil. ASSAY PROCEDURE Internal standard solution: 12 mg/mL of di(2ethylhexyl)phthalate in chloroform Standard solution: Transfer 100 mg of castor oil to a 100mL boiling flask equipped with a suitable reflux condenser connected by a ground-glass joint. Add 30 mL of a mixture of 300 mL of methanol and 3.7 mL of sulfuric acid, reflux in a water bath maintained at 7580 for 2.5 h, cool, and rinse down the condenser with 10 mL of water. Transfer the contents of the flask to a 125-mL separator with the aid of 10 mL of water. Rinse the condenser and the flask with 25 mL of solvent hexane, and transfer to the separator. Shake the separator for 2 min, and draw off the aqueous layer into a second 125-mL separator. Add 20 mL of solvent hexane to the second separator, shake for 2 min, discard the aqueous layer, and transfer the solvent hexane layer to the first separator with the aid of 10 mL of solvent hexane. Wash the combined extracts with three 5-mL portions of water, discarding the washings, and transfer the washed extract to a 125-mL conical flask, through a funnel containing anhydrous sodium sulfate, with the aid of 25 mL of solvent hexane. Place the flask in a hot water bath, and evaporate with the aid of a current of air to dryness. To the residue, add 10.0 mL of Internal standard solution, and mix until solution is complete. Sample solution: Transfer an amount of Emulsion, wellshaken and nominally equivalent to 100 mg of castor oil, to a long-neck, round-bottom, 100-mL boiling flask equipped with a suitable reflux condenser connected by a groundglass joint. Proceed as directed under Standard solution, beginning with Add 30 mL of a mixture of 300 mL of methanol and 3.7 mL of sulfuric acid. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.8-m 4-mm; packed with 4% liquid phase G25 on support S1 Column conditioning: Flush with helium for 25 min, then heat without further flushing at 250 for NLT 30 min, then cool to room temperature, and finally heat while helium is flowing through it at 250 for NLT 60 min.
SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.9570.961 Sample: Capsule contents FATS AND FIXED OILS, Hydroxyl Value 401 Sample solution: Transfer 2 g from Capsule contents to a glass-stoppered, 250-mL conical flask. Add 5.0 mL of a freshly prepared mixture of 1 volume of acetic anhydride and 3 volumes of pyridine, and swirl to mix. Analysis: Connect the flask to a reflux condenser, and heat on a steam bath for 2 h. Add 10 mL of water through the condenser, swirl to mix, heat on a steam bath for an additional 10 min, and allow to cool to room temperature. Add through the condenser 15 mL of normal butyl alcohol that previously has been neutralized to phenolphthalein, remove the condenser, and wash the tip of the condenser and the sides of the flask with an additional 10 mL of neutralized normal butyl alcohol. Add 1 mL of phenolphthalein TS, and titrate with 0.5 N alcoholic potassium hydroxide VS to a faint pink endpoint. Perform a blank determination on a 5.0 mL portion of the acetic anhydridepyridine mixture. To determine the amount of free acid in the Oil, weigh accurately 10 g into a 250-mL conical flask, add 10 mL of pyridine that previously has been neutralized to phenolphthalein, swirl to mix, add 1 mL of phenolphthalein TS, and titrate with 0.5 N alcoholic potassium hydroxide VS to a faint pink endpoint. Calculate the hydroxyl value taken: Result = 56.1N[A + (BW/D) C]/W = normality determined for the alcoholic potassium hydroxide solution A = volume of 0.5 N alcoholic potassium hydroxide consumed by the blank (mL) B = volume consumed in the free-acid titration (mL) W = weight of Oil taken (g) D = weight of Oil used in the free-acid titration (g) C = volume consumed in the sample titration (mL) Acceptance criteria: 160168 FATS AND FIXED OILS, Iodine Value 401: 8388 Sample: Capsule contents FATS AND FIXED OILS, Saponification Value 401: 176182 Sample: Capsule contents ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, preferably at a controlled room temperature. N
USP 32
Temperature Column: 245 Injection port: 300 Detector: 300 Flow rate: Adjust to obtain a peak due to castor oil 5.5 min after introduction of the specimen and an internal standard peak 8 min after introduction of the specimen. Carrier gas: Helium Injection size: 5 L Analysis Sample: Standard solution and Sample solution Measure the heights of the peaks due to castor oil and internal standard Calculate the percentage of castor oil in the portion of Emulsion: Result = (RU/RS) (CS/CU) 100 = ratio of the heights of the peaks due to castor oil and internal standard from the Sample solution = ratio of the heights of the peaks due to castor oil RS and internal standard from the Standard solution = concentration of castor oil in the Standard CS solution (mg/mL) = nominal concentration of castor oil in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%120.0% RU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Official Monographs / Cefaclor 105

solution, beginning with Add 30 mL of a mixture of 300 mL of methanol and 3.7 mL of sulfuric acid Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.8-m 4-mm column packed with 4% liquid phase G25 on support S1 Column conditioning: Flush with helium for 25 min, then heat without further flushing at 250 for NLT 30 min, then cool to room temperature, and finally heat while helium is flowing through it at 250 for NLT 60 min. Temperature Column: 245 Injection port: 300 Detector: 300 Flow rate: Adjust to obtain a peak due to castor oil 5.5 min after introduction of the specimen and an internal standard peak 8 min after introduction of the specimen. Carrier gas: Helium Injection size: 5 L Analysis Sample: Standard solution and Sample solution Measure the heights of the peaks due to castor oil and Internal standard solution. Calculate the percentage, of castor oil in the portion of Aromatic Castor Oil: Result = (RU/RS) (Cs/CU) 100 = ratio of the heights of the peaks due to castor oil and internal standard from the Sample solution = ratio of the heights of the peaks due to castor oil RS and internal standard from the Standard solution = concentration of the Standard solution (mg/mL) CS = nominal concentration of castor oil in the Sample CU solution (mg/mL) Acceptance criteria: NLT 95.0% OTHER COMPONENTS ALCOHOL DETERMINATION, Method I 611: C2H5OH NMT 4.0% of RU
Aromatic Castor Oil

(Comment on this Monograph)id=m13830=Aromatic Castor Oil=Ca-Chl-Monos.pdf) DEFINITION Aromatic Castor Oil is Castor Oil containing suitable flavoring agents. It contains NLT 95.0% of castor oil. ASSAY PROCEDURE Internal standard solution: 12 mg/mL of di(2ethylhexyl)phthalate in chloroform Standard solution: Transfer 100 mg of castor oil to a 100mL boiling flask equipped with a suitable reflux condenser connected by a ground-glass joint. Add 30 mL of a mixture of 300 mL of methanol and 3.7 mL of sulfuric acid, reflux in a water bath maintained at 7580 for 2.5 h, cool, and rinse down the condenser with 10 mL of water. Transfer the contents of the flask to a 125-mL separator with the aid of 10 mL of water. Rinse the condenser and the flask with 25 mL of solvent hexane, and transfer to the separator. Shake the separator for 2 min, and draw off the aqueous layer into a second 125-mL separator. Add 20 mL of solvent hexane to the second separator, shake for 2 min, discard the aqueous layer, and transfer the solvent hexane layer to the first separator with the aid of 10 mL of solvent hexane. Wash the combined extracts with three 5-mL portions of water, discarding the washings, and transfer the washed extract to a 125-mL conical flask, through a funnel containing anhydrous sodium sulfate, with the aid of 25 mL of solvent hexane. Place the flask in a hot water bath, and evaporate with the aid of a current of air to dryness. To the residue, add 10.0 mL of Internal standard solution, and mix until solution is complete. Sample solution: Transfer 100 mg of Aromatic Castor Oil to a long-neck, round-bottom, 100-mL boiling flask equipped with a suitable reflux condenser connected by a ground-glass joint. Proceed as directed under Standard
Cefaclor
(Comment on this Monograph)id=m13880=Cefaclor=Ca-ChlMonos.pdf)
385.82 C15H14ClN3O4S H2O 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7[(aminophenylacetyl)amino]-3-chloro-8-oxo-, monohydrate, [6R-[6,-7 (R*)]]-; (6R,7R)-7-[(R)-2-Amino-2-phenylacetamido]-3-chloro-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid monohydrate; 3-Chloro-7-D-(2-phenylglycinamido)-3-cephem-4-carboxylic acid monohydrate [70356-03-5]; Anhydrous 367.81 [53994-73-3].
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Cefaclor / Official Monographs

Mobile Phase: See Gradient Table.
Gradient Table Time (min) 0 30 45 55 60 70 Solution A (%) 95 75 0 0 95 95
USP 32
DEFINITION Cefaclor has a potency of NLT 950 g and NMT 1020 g of C15H14ClN3O4S/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time for cefaclor of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate in a mixture of 780 mL of water and 10 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1, and add 220 mL of methanol System suitability solution: 0.3 mg/mL of cefaclor and 0.3 mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile phase [NOTESonicate briefly, if necessary, to achieve dissolution, and avoid heating the solution. Use within 8 h if stored at room temperature or within 20 h if stored under refrigeration.] Sample solution: 0.3 mg/mL of Cefaclor in Mobile phase [NOTESonicate briefly, if necessary, to achieve dissolution, and avoid heating the solution. Use within 8 h if stored at room temperature, or within 20 h if stored under refrigeration.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for cefaclor and cefaclor, delta-3 isomer are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 Between the cefaclor peak and the cefaclor, delta-3 isomer peak Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the potency, in g/mg, of cefaclor C15H14ClN3O4S in each mg of the Cefaclor taken: Result = (rU/rS) (CS/CU) P = peak area of Sample solution = peak area of the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = concentration of Cefaclor in the Sample solution CU (mg/mL) P = designated potency, in g of (C15H14ClN3O4S)/mg, of USP Cefaclor RS Acceptance criteria: 950 g/mg1020 g/mg IMPURITIES Organic Impurities PROCEDURE Diluent: 2.4 mg/mL of monobasic sodium phosphate in water, adjust with phosphoric acid to a pH of 2.5 Blank solution: Use the Diluent. Solution A: 6.9 mg/mL of monobasic sodium phosphate in water; adjust with phosphoric acid to a pH of 4.0 Solution B: Acetonitrile and Solution A (9:11), degassing for NMT 2 min rU rS CS
Solution B (%) 5 25 100 100 5 5
Standard solution: 0.05 mg/mL of USP Cefaclor RS in Diluent [NOTESonicate briefly, if necessary, to dissolve, and avoid heating. Use this solution on the day it is prepared.] System suitability solution: 0.05 mg/mL of USP Cefaclor, Delta-3 Isomer RS dissolved in Standard solution Sample solutions: Transfer 50 mg of Cefaclor into each of two 10-mL volumetric flasks, and dilute each with Diluent to volume. Sonicate briefly, if necessary, to dissolve, and avoid heating. [NOTEUse these within 2 h when stored at room temperature or within 20 h when stored under refrigeration.] Chromatographic system (See Chromatography 621, System Suitability.) [NOTEReducing the acetonitrile content increases the retention time of cefaclor and increases the resolution between cefaclor, delta-3 isomer and cefaclor.] Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Blank solution [NOTEThe retention time for the cefaclor peak is 2329 min]. Suitability requirements Resolution: NLT 2.0 between cefaclor, delta-3 isomer and cefaclor, System suitability solution Tailing factor: NMT 1.2 for the cefaclor peak, System suitability solution [NOTEExamine the chromatogram of the Blank solution for any extraneous peaks, and disregard any corresponding peaks observed in the chromatogram of the Sample solutions. Ensure that any extraneous peaks observed do not represent carryover from previous injections.] Analysis Samples: Standard solution and Sample solutions Calculate the percentage of each cefaclor related compound in the portion of Cefaclor taken: Result = (ri/rS) (CS/CU) P F 100 ri rS CS CU P F = peak area of an individual related compound of the Sample solution = peak area of the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = concentration of Cefaclor in the respective Sample solution (mg/mL) = designated potency of USP Cefaclor RS (g/mg) = conversion factor, 0.001 mg/g
USP 32
Acceptance criteria Individual Impurities: Determine the mean values for each cefaclor related compound: NMT 0.5% of any individual cefaclor related compound is found. Total impurities: NMT 2.0% of total cefaclor related compounds is found. [NOTEIn an acceptable determination, the difference between duplicate determinations of total cefaclor related compounds is NMT 0.2% absolute, or the variation from the mean of the two values is NMT 10%, whichever is greater.] SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 3.04.5, in an aqueous suspension containing 25 mg/mL WATER DETERMINATION, Method I 921: 3.0%6.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefaclor RS USP Cefaclor, Delta-3 Isomer RS

Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for cefaclor and cefaclor, delta-3 isomer are 0.8 and 1.0, respectively. ] Suitability requirements Resolution: NLT 2.5 between the cefaclor peak and the cefaclor, delta-3 isomer peak Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the potency, in g/mg, cefaclor C15H14ClN3O4S in the portion of Cefaclor Capsules taken: Result = (rU/rS) (CS /CU) P = peak area of the Sample solution = peak area of the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = nominal concentration of Cefaclor in the Sample CU solution (mg/mL) P = designated potency of USP Cefaclor RS (g of C15H14ClN3O4S/mg ) Acceptance criteria: 90.0%120% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Sample solutions: Sample per Dissolution 711. Dilute with Medium as needed, and filter. Standard solution: USP Cefaclor RS of a known concentration in Medium Analysis: Determine the amount of C15H14ClN3O4S dissolved from UV absorbances at 264 nm of the Sample solution, in comparison with the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of cefaclor (C15H14ClN3O4S) is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Diluent: 2.4 mg/mL of monobasic sodium phosphate in water; adjusted with phosphoric acid to a pH of 2.5 Blank solution: Use the Diluent. Solution A: Dissolve 6.9 g of monobasic sodium phosphate in 1000 mL of water, and adjust with phosphoric acid to a pH of 4.0. Solution B: Acetonitrile and Solution A (9:11), degassing for NMT 2 min rU rS CS
Cefaclor Capsules
(Comment on this Monograph)id=m13890=Cefaclor Capsules=Ca-Chl-Monos.pdf) DEFINITION Cefaclor Capsules contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of C15H14ClN3O4S. IDENTIFICATION PROCEDURE Sample solution: Mix the contents of 1 capsule with water and filter to obtain a concentration of 2 mg/mL. Analysis: Proceed as directed in the Assay. Acceptance criteria: The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate in a mixture of 780 mL of water and 10 mL of triethylamine A. Adjust with phosphoric acid to a pH of 2.5 0.1, and add 220 mL of methanol. System suitability solution: 0.3 mg/mL of cefaclor and 0.3 mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile phase [NOTESonicate briefly, if necessary, to achieve dissolution, and avoid heating the solution. Use within 8 h if stored at room temperature, or within 20 h if stored under refrigeration.] Sample solution: Remove the contents of NLT 20 Cefaclor Capsules as completely as possible, and weigh. Mix the combined contents, and transfer a portion of the powder, nominally equivalent to 75 mg of cefaclor, to a 250-mL volumetric flask, dilute with Mobile phase to volume, and mix. Sonicate if necessary to ensure complete dissolution of the cefaclor. Filter to obtain the clear Sample solution.
108

See Gradient Table.
Gradient Table Time 0 30 45 55 60 70 Solution A (%) 95 75 0 0 95 95 Solution B (%) 5 25 100 100 5 5
USP 32
F = conversion factor (0.001 mg/g) Acceptance criteria Individual impurities: NMT 0.5% of any individual cefaclor-related compound is found. Total impurities: NMT 2.0% of all cefaclor-related compounds, not including the contribution of any peak that gives a result of less than 0.1% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 8.0%
Mobile Phase:
Standard solution: 0.05 mg/mL of USP Cefaclor RS in Diluent [NOTESonicate briefly, if necessary to dissolve, and avoid heating. Use this solution on the day it is prepared.] System suitability solution: 0.05 mg/mL of USP Cefaclor, Delta-3 Isomer RS dissolved in the Standard solution Sample solution: Remove the contents of NLT 20 Cefaclor Capsules as completely as possible, and mix. Transfer a portion of the combined contents, equivalent to about 50 mg of cefaclor, to a 10-mL volumetric flask. Dissolve in Diluent, using brief sonication, if necessary, to achieve dissolution. Avoid heating. Dilute with Diluent to volume, mix, and filter. Use within 3 h if stored at room temperature, or within 20 h when stored under refrigeration. Chromatographic system (See Chromatography 621, System Suitability.) [NOTEReducing the acetonitrile content increases the retention time of cefaclor and increases the resolution between cefaclor, delta-3 isomer and cefaclor.] Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Blank solution [NOTEThe retention time for the cefaclor peak is 2329 min.] Suitability requirements Resolution: NLT 2.0 between cefaclor, delta-3 isomer and cefaclor, System suitability solution Tailing factor: NMT 1.2 for the cefaclor peak, System suitability solution [NOTEExamine the chromatogram of the Blank solution for any extraneous peaks, and disregard any corresponding peaks in the Sample solutions. Ensure that any extraneous peaks observed do not represent carryover from previous injections.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of each related compound in the portion of Cefaclor Capsules taken: Result = (rU/rS) (CS/CU) P F 100 rU rS CS CU P = peak area of the Sample solution = peak area of the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = nominal concentration of cefaclor in the Sample solution (mg/mL) = potency of USP Cefaclor RS (g/mg of C15H14ClN3O4S)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefaclor RS USP Cefaclor, Delta-3 Isomer RS
Cefaclor for Oral Suspension

(Comment on this Monograph)id=m13900=Cefaclor for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cefaclor for Oral Suspension is a dry mixture of Cefaclor and one or more suitable buffers, colors, diluents, and flavors. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of C15H14ClN3O4S. IDENTIFICATION The retention time of the major peak for cefaclor in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate in a mixture of 780 mL of water and 10 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1, and add 220 mL of methanol. System suitability solution: 0.3 mg/mL of cefaclor and 0.3 mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile phase [NOTESonicate briefly, if necessary, to achieve dissolution, and avoid heating the solution. Use within 8 h if stored at room temperature, or within 20 h if stored under refrigeration.] Sample stock solution: Constitute Cefaclor for Oral Suspension as directed in the labeling, freshly mixed and free from air bubbles. Sample solution: 0.3 mg/mL of cefaclor from Sample stock solution diluted with Mobile phase. Sonicate if necessary to ensure complete dissolution of the cefaclor, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for cefaclor and cefaclor, delta-3 isomer are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between the cefaclor and cefaclor, delta-3 isomer peaks
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Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the potency, in mg, of C15H14ClN3O4S in the portion of the constituted Cefaclor for Oral Suspension taken: Result = (rU/rS) (CS/CU) P F 100 = peak area of the cefaclor peak from the Sample solution = peak area of the cefaclor peak from the Standard rS solution = concentration of USP Cefaclor RS in the Standard CS solution (mg/mL) = nominal concentration of cefaclor in the Sample CU solution (mg/mL) P = potency of USP Cefaclor RS (mg) F = conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120% rU PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905 For solid packaged in single-unit containers: Meets the requirements IMPURITIES Organic Impurities PROCEDURE Diluent: 2.4 mg/mL of monobasic sodium phosphate in water, adjust with phosphoric acid to a pH of 2.5 Blank solution: Use the Diluent. Solution A: Dissolve 6.9 g of monobasic sodium phosphate in 1000 mL of water, and adjust with phosphoric acid to a pH of 4.0. Solution B: Acetonitrile and Solution A (9:11), degassing for NMT 2 min Mobile Phase: See Gradient Table.
Gradient Table Time (min) 0 30 45 55 60 70 Solution A (%) 95 75 0 0 95 95 Solution B (%) 5 25 100 100 5 5

Chromatographic system (See Chromatography 621, System Suitability.) [NOTEReducing the acetonitrile content increases the retention time of cefaclor and increases the resolution between cefaclor, delta-3 isomer and cefaclor.] Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Blank solution [NOTEThe retention time for the cefaclor peak is 2329 min.] Suitability requirements Resolution: NLT 2.0 between cefaclor, delta-3 isomer and cefaclor, System suitability solution Tailing factor: NMT 1.2 for the cefaclor peak, System suitability solution [NOTEExamine the chromatogram of the Blank solution for any extraneous peaks, and disregard any corresponding peaks observed in the chromatogram of the Sample solutions. Ensure that any extraneous peaks observed do not represent carryover from previous injections.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of each related compound in the portion of Cefaclor for Oral Suspension taken: Result = (rU/rS) (CS/CU) P F 100 = peak response area of an individual related compound from the Sample solution = peak response area from the Standard solution rS = concentration of USP Cefaclor RS in the CS Standard solution (mg/mL) = nominal concentration of cefaclor in the Sample CU solution (mg/mL) P = designated potency of USP Cefaclor RS (g/mg) F = conversion factor, 0.001 mg/g Acceptance criteria Individual impurities: NMT 1.0% of any individual cefaclor-related compound is found. Total impurities: NMT 3.0% of all cefaclor-related compounds, not including the contribution of any peak that gives a result of less than 0.1%. rU SPECIFIC TESTS PH 791: 2.55.0, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefaclor RS USP Cefaclor, Delta-3 Isomer RS
Standard solution: 0.05 mg/mL of USP Cefaclor RS in Diluent [NOTESonicate briefly, if necessary, to dissolve, and avoid heating. Use this solution on the day it is prepared.] System suitability solution: 0.05 mg/mL of USP Cefaclor, Delta-3 Isomer RS dissolved in Standard solution Sample solution: Constitute Cefaclor for Oral Suspension as directed in the labeling. Transfer a portion of Cefaclor for Oral Suspension, freshly mixed and free from air bubbles, equivalent to 50 mg of cefaclor, to a 10-mL volumetric flask. Dissolve in Diluent, using brief sonication, if necessary, to achieve dissolution. Avoid heating. Dilute with Diluent to volume, and filter. Use within 3 h if stored at room temperature, or within 20 h when stored under refrigeration.
Cefaclor Chewable Tablets

(Comment on this Monograph)id=m662=Cefaclor Chewable Tablets=Ca-Chl-Monos.pdf) DEFINITION Cefaclor Chewable Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of cefaclor (C15H14ClN3O4 S).
110

AS CS
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= absorbance from the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = nominal concentration of cefaclor in the Sample CU solution (mg/mL) Tolerances: NLT 80% (Q) of the labeled amount of cefaclor UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Solvent: 2.4 mg/mL of monobasic sodium phosphate in water; adjust with phosphoric acid to a pH of 2.5 Blank: Use the Solvent. Solution A: 6.9 mg/mL of monobasic sodium phosphate in water; Adjust with phosphoric acid to a pH of 4.0 Solution B: Acetonitrile and Solution A (9:11), degassing for NMT 2 min Mobile Phase: See Gradient Table.
IDENTIFICATION The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate in a mixture of 780 mL of water and 10 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1, and add 220 mL of methanol. System suitability solution: 0.3 mg/mL of cefaclor and 0.3 mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Standard solution: 0.3 mg/mL of USP Cefaclor RS in the Mobile phase. [NOTESonicate briefly, if necessary, to achieve dissolution, and avoid heating the solution. Use this Standard solution within 8 h if stored at room temperature, or within 20 h if stored under refrigeration.] Sample solution: Transfer an equivalent to 75 mg of cefaclor, from weighed and finely powdered Chewable Tablets (NLT 20), to a 250-mL volumetric flask, and dilute with Mobile phase to volume. Sonicate, if necessary, to dissolve the cefaclor. Filter to obtain a clear solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention time for cefaclor and cefaclor delta-3 isomer are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between the cefaclor peak and the cefaclor, delta-3 isomer peak Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of cefaclor C15H14ClN3O4S in the portion of Chewable Tablets taken: Result = (rU/rS) (CS/CU) 100 = cefaclor peak area rom the Sample solution = cefaclor peak area from the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = nominal concentration of cefaclor in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Sample solution: Sample per Dissolution 711 Analysis: Determine the amount of cefaclor dissolved by employing UV absorption at the wavelength of maximum absorption at about 264 nm on filtered portions of the Sample solution, suitably diluted with Medium, if necessary, in comparison with a Standard solution having a known concentration of USP Cefaclor RS in the same Medium. Calculate the amount of cefaclor dissolved: Result = (AU/AS) (CS/CU) 100 AU = absorbance from the Sample solution rU rS CS
System suitability solution: 0.05 mg/mL of USP Cefaclor, Delta-3 Isomer RS in the Standard solution Standard solution: 0.05 mg/mL of USP Cefaclor RS in the Solvent [NOTESonicate briefly, if necessary, to dissolve, and avoid heating. Use this solution on the day it is prepared.] Sample solution: Transfer an equivalent to 50 mg of cefaclor, from weighed and finely powdered Chewable Tablets (NLT 20), to a 10-mL volumetric flask. Dissolve in Solvent, using brief sonication, if necessary, to dissolve. Avoid heating. Dilute with Solvent to volume and filter. [NOTEUse this Sample solution within 3 h if stored at room temperature, or within 20 h when stored under refrigeration.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe retention time for cefaclor is 2329 min.] Suitability requirements Resolution: NLT 2.0 between cefaclor delta-3 isomer and cefaclor Tailing factor: NMT 1.2 [NOTEExamine the chromatogram for any extraneous peaks, and disregard any corresponding peaks observed in the chromatogram of the Sample solution. Ensure that any extraneous peaks observed do not represent carryover from previous injections.]
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of each cefaclor related compound in the portion of Cefaclor: Result = (ri/rs) (CS/CU) 100 = peak response of an individual related compound in the chromatogram from the Sample solution = peak response for the cefaclor peak in the rS chromatogram of the Standard solution = concentration of USP Cefaclor RS in the Standard CS solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria Any individual related compound: NMT 1.0% Total impurities: NMT 3.0%, not including the contribution of any peak that gives a result of less than 0.1% [NOTEIn an acceptable determination, the difference between duplicate determinations of total cefaclor related compounds NMT 0.2% absolute, or the variation from the mean of the two values NMT 10%, whichever is greater.] ri SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 5.0%

volumetric flask, and dilute with Mobile phase to volume. Sonicate, if necessary, to dissolve the cefaclor. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for cefaclor and cefaclor, delta-3 isomer are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between the cefaclor peak and the cefaclor, delta-3 isomer peak Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of label claim of C15H14ClN3O4S in the portion of Cefaclor Extended Release Tablets taken: Result = (rU/rS) (CS/CU) P F 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = nominal concentration of Cefaclor in the Sample CU solution (mg/mL) P = designated potency of USP Cefaclor RS [g of cefaclor (C15H14ClN3O4S)/mg] F = conversion factor, 0.001 mg/g Acceptance criteria: 90.0%110% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1 (10-mesh basket): 100 rpm Time: 30, 60, and 240 min Standard solution: 25 g/mL USP Cefaclor RS in Medium Sample solutions: Dilute filtered portions of the solution under test with Medium to obtain solutions having an estimated concentration of 25 g/mL cefaclor. Analysis: Determine the amount of cefaclor (C15H14ClN3O4S) dissolved in the Sample solution by employing UV absorption at 265 nm, in comparison with the Standard solution. Tolerances: The percentages of the labeled amount of cefaclor (C15H14ClN3O4S) dissolved at the times specified conform to Acceptance Table 2.
Time (min) 30 60 240 Amount dissolved Between 5% and 30% Between 20% and 50% NLT 80%
rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25, excursions permitted between 15 and 30. LABELING: The product label and product labeling indicate that the Chewable Tablets must be chewed or crushed before administration. USP REFERENCE STANDARDS 11 USP Cefaclor RS USP Cefaclor Delta-3 Isomer RS
Cefaclor Extended-Release Tablets

(Comment on this Monograph)id=m13905=Cefaclor ExtendedRelease Tablets=Ca-Chl-Monos.pdf) DEFINITION Cefaclor Extended-Release Tablets contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of cefaclor (C15H14ClN3O4S). IDENTIFICATION The retention time for cefaclor from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 1 g of sodium 1-pentanesulfonate in a mixture of 780 mL of water and 10 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1, add 220 mL of methanol. System suitability solution: 0.3 mg/mL of cefaclor and 0.3 mg/mL of USP Cefaclor, Delta-3 Isomer RS in Mobile phase Standard solution: 0.3 mg/mL of USP Cefaclor RS in Mobile phase [NOTESonicate briefly, if necessary, to achieve dissolution, and avoid heating the solution. Use within 8 h if stored at room temperature, or within 20 h if stored under refrigeration.] Sample solution: Weigh and finely powder NLT 20 Cefaclor Extended Release Tablets. Transfer a portion of the powder, nominally equivalent to 75 mg of cefaclor, to a 250-mL
IMPURITIES Organic Impurities PROCEDURE Diluent: 2.4 mg/mL of monobasic sodium phosphate in water; adjusted with phosphoric acid to a pH of 2.5 Blank solution: Use the Diluent. Solution A: Dissolve 6.9 g of monobasic sodium phosphate in 1000 mL of water, and adjust with phosphoric acid to a pH of 4.0. Solution B: Acetonitrile and Solution A (9:11), degassing for NMT 2 min
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See Gradient Table.
USP 32
Acceptance criteria Individual impurities: NMT 0.6% of any individual cefaclor-related compound is found. Total impurities: NMT 2.0% of all cefaclor-related compounds, not including the contribution of any peak that gives a result of less than 0.1% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 7.0%
Mobile Phase:
Standard solution: 0.05 mg/mL of USP Cefaclor RS in Diluent [NOTESonicate briefly, if necessary, to dissolve, and avoid heating. Use this solution on the day it is prepared.] System suitability solution: 0.05 mg/mL of USP Cefaclor, Delta-3 Isomer RS dissolved in Standard solution Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, nominally equivalent to 50 mg of cefaclor, to a 10-mL volumetric flask. Dissolve in Diluent, using brief sonication, if necessary, to achieve dissolution. Avoid heating. Dilute with Diluent to volume, mix, and filter. [NOTEUse within 3 h if stored at room temperature, or within 20 h if stored under refrigeration.] Chromatographic system (See Chromatography 621, System Suitability.) [NOTEReducing the acetonitrile content increases the retention time of cefaclor and increases the resolution between cefaclor, delta-3 isomer and cefaclor.] Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Blank solution and System suitability solution [NOTEThe retention time for the cefaclor peak is 2329 min.] Suitability requirements Resolution: NLT 2.0 between cefaclor, delta-3 isomer and cefaclor, System suitability solution Tailing factor: NMT 1.2 for the cefaclor peak, System suitability solution [NOTEExamine the chromatogram of the Blank solution for any extraneous peaks, and disregard any corresponding peaks observed in the chromatogram of the Sample solutions. Ensure that any extraneous peaks observed do not represent carryover from previous injections.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of each related compound in the portion of Cefaclor Extended Release Tablets taken: Result = (rU/rS) (CS/CU) P F 100 rU rS CS CU P F = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Cefaclor RS in the Standard solution (mg/mL) = nominal concentration of cefaclor in the Sample solution (mg/mL) = potency of USP Cefaclor RS [g of cefaclor (C15H14ClN3O4S)/mg] = conversion factor, 0.001 mg/g
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cefaclor RS USP Cefaclor, Delta-3 Isomer RS
Cefadroxil
(Comment on this Monograph)id=m13930=Cefadroxil=Ca-ChlMonos.pdf)
381.40 C16H17N3O5S H2O 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[ [amino(4-hydroxyphenyl)acetyl]amino]-3-methyl-8-oxo-, monohydrate, [6R-[6,7 (R*)]]-; (6R,7R)-7-[(R)-2-Amino-2-(p-hydroxyphenyl)acetamido]-3methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid monohydrate [66592-87-8]; Hemihydrate 372.39 [119922-85-9]; Anhydrous 363.40 [50370-12-2]. DEFINITION Cefadroxil has a potency equivalent to NLT 950 g and NMT 1050 g of C16H17N3O5S/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 2 mg/mL of USP Cefadroxil RS Sample solution: 2 mg/mL of Cefadroxil Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 20 L Pre-developing solvent solution: n-hexane and tetradecane (95:5) Developing solvent system: 0.1M citric acid, 0.1M dibasic sodium phosphate and a 1 in 15 solution of ninhydrin in acetone (60:40:15) Spray reagent: 1in500 solution of ninhydrin in dehydrated alcohol [NOTEProtect Spray reagent from light.] Analysis Samples: Standard solution and Sample solution Place the thin-layer chromatographic plate in a chamber containing the Pre-developing solvent solution and allow the solvent front to move the length of the plate. Remove the plate from the chamber and allow the solvent to evaporate. Apply the Sample solution and Standard solution
USP 32
to the plate, allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow to air-dry. Spray the plate with Spray reagent, dry for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Buffer solution: Dissolve 13.6 g of monobasic potassium phosphate in water to make 2000 mL of solution. Adjust with 10 N potassium hydroxide to a pH of 5.0. Mobile phase: Acetonitrile and Buffer solution (1:24) Standard solution: 1.06 mg/mL of USP Cefadroxil RS in Buffer solution [NOTEThis solution contains the equivalent of 1000 g/ mL of C16H17N3O5S. Use this solution on the day prepared.] Sample solution: 1.06 mg/mL of Cefadroxil in Buffer solution [NOTEStir by mechanical means for 5 min until dissolved. Use this solution on the day prepared.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: Between 2.0 and 3.5 Column efficiency: NLT 1800 theoretical plates Tailing factor: NMT 2.2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C16H17N3O5S in each mg of Cefadroxil taken: Result = (rU/rS) (CS/CU) P = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cefadroxil RS in the Standard solution (mg/mL) = concentration of Cefadroxil taken to prepare the CU Sample solution (mg/mL) P = cefadroxil equivalent (g/mg) of USP Cefadroxil RS Acceptance criteria: 9501050 g/mg rU rS CS IMPURITIES Organic Impurities PROCEDURE 1 Diluent: Alcohol, 2.4 N hydrochloric acid, and water (75:3:22) Standard solution A: Dilute 1.0 mL of the Sample solution to 100 mL with Diluent. Standard solution B: 0.25 mg/mL each of 7aminodesacetoxycephalosporanic acid and D--4hydroxyphenylglycine in Diluent Standard solution C: 0.25 mg/mL of D--4hydroxyphenylglycine in Solution A Sample solution: 25 mg/mL of Cefadroxil in Diluent Identification solution: Standard solution B and Sample solution (1:1)
Official Monographs / Cefadroxil 113

Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 2-L portions of the Sample solution, Standard solution A, Standard solution B, and Standard solution C, and a 4-L portion of the Identification solution Developing solvent system: Ethyl acetate, alcohol, formic acid, and water (14:5:1:5) Analysis Samples: Sample solution, Standard solution A, Standard solution B, Standard solution C, and Identification solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatograms until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the plate to dry, and examine the chromatograms under shortwavelength UV light. [NOTEIn a valid test the chromatogram obtained from the Identification solution shows three clearly separated spots.] Acceptance criteria: Any secondary spot in the chromatogram of the Sample solution corresponding to 7aminodesacetoxycephalosporanic acid or D--4hydroxyphenylglycine is not more intense than the corresponding spot in the chromatogram of Standard solution B (1.0%); and any spot, other than the principal spot and any spot corresponding to 7aminodesacetoxycephalosporanic acid or D--4hydroxyphenylglycine, is not more intense than the principal spot in the chromatogram of Standard solution A (1.0%). PROCEDURE 2: DIMETHYLANILINE 223: Meets the requirement SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +165.0 to +178.0 Sample solution: 10 mg/mL CRYSTALLINITY 695: Meets the requirements PH 791: 4.06.0, in a suspension containing 50 mg/mL WATER DETERMINATION, Method I 921: 4.2%6.0%, except that where it is labeled as being in the hemihydrate form it is between 2.4% and 4.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The hemihydrate form is so labeled. USP REFERENCE STANDARDS 11 USP Cefadroxil RS
Cefadroxil Capsules
(Comment on this Monograph)id=m13940=Cefadroxil Capsules=Ca-Chl-Monos.pdf) DEFINITION Cefadroxil Capsules contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cefadroxil (C16H17N3O5S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 2 mg/mL of USP Cefadroxil RS Sample solution: 2 mg/mL of cefadroxil, from the contents of 1 Capsule dissolved in water Chromatographic system (See Chromatography 621, System Suitability.)
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Cefadroxil / Official Monographs

CU
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= nominal concentration of cefadroxil in the Sample solution (mg/mL) P = cefadroxil equivalent of USP Cefadroxil RS (g/mg) F = conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 30 min Standard solution: USP Cefadroxil RS in Medium of a known concentration Sample solution: Sample per Dissolution 711. Suitably dilute with Medium, if necessary, and filter. Analysis: Determine the amount of C16H17N3O5S dissolved from UV absorbances at 263 nm of the Sample solution in comparison to the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C16H17N3O5S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 7.0%
Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 20 L Pre-developing solvent solution: n-hexane and tetradecane (95:5) Developing solvent system: 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a 1in15 solution of ninhydrin in acetone (60:40:15) Spray reagent: 1in500 solution of ninhydrin in dehydrated alcohol [NOTEProtect Spray reagent from light.] Analysis Samples: Standard solution and Sample solution Place the thin-layer chromatographic plate in a chamber containing the Pre-developing solvent solution and allow the solvent front to move the length of the plate. Remove the plate from the chamber and allow the solvent to evaporate. Apply the Sample solution and Standard solution to the plate, allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow to air-dry. Spray the plate with the Spray reagent, dry for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Buffer solution: Dissolve 13.6 g of monobasic potassium phosphate in water to make 2000 mL of solution. Adjust with 10 N potassium hydroxide to a pH of 5.0. Mobile phase: Acetonitrile and Buffer solution (1:24) Standard solution: 1.06 mg/mL of USP Cefadroxil RS in Buffer solution [NOTEThis solution contains the equivalent of 1000 g/mL of C16H17N3O5S. Use this solution on the day prepared.] Sample solution: Remove, as completely as possible, the contents of NLT 10 Capsules, and weigh. Transfer a portion of the powder, nominally equivalent to 200 mg of cefadroxil, to a 200-mL volumetric flask. Dilute with Buffer solution to volume, and stir by mechanical means for 5 min. [NOTEUse this solution on the day prepared.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: 2.03.5 Column efficiency: NLT 1800 theoretical plates Tailing factor: NMT 2.2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O5S in the portion of Capsules taken: Result = (rU/rS) (CS/CU) P F 100 rU rS CS = peak response of cefadroxil from the Sample solution = peak response of cefadroxil from the Standard solution = concentration of USP Cefadroxil RS in the Standard solution (mg/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Capsules prepared using the hemihydrate form of Cefadroxil are so labeled. USP REFERENCE STANDARDS 11 USP Cefadroxil RS
Cefadroxil for Oral Suspension

(Comment on this Monograph)id=m13950=Cefadroxil for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cefadroxil for Oral Suspension is a dry mixture of Cefadroxil and one or more suitable buffers, colors, diluents, and flavors. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of C16H17N3O5S. IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 2 mg/mL of USP Cefadroxil RS Sample solution: Constitute 1 container of Cefadroxil for Oral Suspension as directed in the labeling. Dilute a portion of the resulting suspension to a concentration of 2 mg/mL. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 20 L Pre-developing solvent solution: n-hexane and tetradecane (95:5) Developing solvent system: 0.1M citric acid, 0.1M dibasic sodium phosphate and a 1 in 15 solution of ninhydrin in acetone (60:40:15) Spray reagent: 1in500 solution of ninhydrin in dehydrated alcohol [NOTEProtect Spray reagent from light.] Analysis Samples: Standard solution and Sample solution Place the thin-layer chromatographic plate in a chamber containing the Pre-Developing solvent solution and allow the solvent front to move the length of the plate. Remove the plate from the chamber and allow the solvent to evaporate. Apply the Sample solution and Standard solution to the plate, allow the spots to dry, and develop the
USP 32
chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow to air-dry. Spray the plate with the Spray reagent, dry for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Buffer solution: Dissolve 13.6 g of monobasic potassium phosphate in water to make 2000 mL of solution. Adjust with 10 N potassium hydroxide to a pH of 5.0 Mobile phase: Acetonitrile and Buffer solution (1:24) Standard solution: 1.06 mg/mL of USP Cefadroxil RS in Buffer solution [NOTEThis solution contains the equivalent of 1000 g/ mL of cefadroxil (C16H17N3O5S). Use this solution on the day prepared.] Sample solution: Constitute a container of Cefadroxil for Oral Suspension as directed in the labeling. Dilute a portion of the resulting suspension with Buffer solution to prepare a solution containing nominally 1.0 mg/mL. Pass through a suitable filter of 0.8m or finer porosity, and use the filtrate. Use this solution on the day prepared. Chromatographic system (See Chromatography 621, System suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k : Between 2.0 and 3.5 Column efficiency: NLT 1800 theoretical plates Tailing factor: NMT 2.2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O5S of label claim in Cefadroxil for Oral Suspension: Result = (rU/rS) (CS/CU) P F 100 = peak response of the cefadroxil of the Sample solution rS = peak response of the cefadroxil of the Standard solution CS = concentration of USP Cefadroxil RS in the Standard solution (mg/mL) CU = nominal concentration of cefadroxil in the Sample solution (mg/mL) P = cefadroxil equivalent (g/mg) of USP Cefadroxil RS F = conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120% rU PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 25 rpm Time: 30 min Standard solution: USP Cefadroxil RS in Medium at a known concentration Sample solution: Transfer 5.0 mL of the constitued Oral Suspension (weighed) to the dissolution vessel.
Official Monographs / Cefadroxil 115

Analysis Determine the amount of cefadroxil dissolved by employing UV absorption at the wavelength of 263 nm on the Sample solution in comparison with the Standard solution Calculate the amount of cefadroxil dissolved: Result = (AU/AS) (Cs/W) (V/D) 100 = absorbance of the Sample solution AU = absorbance of the Standard solution AS = concentration of Standard solution (mg/mL) CS W = weight of Sample (mg) V = volume of Medium (mL), 900 D = dilution factor Tolerances: NLT 75% (Q) of the labeled amount of cefadroxil is dissolved. UNIFORMITY OF DOSAGE UNITS 905: For solid packaged in single-unit containers: Meets the requirements DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 4.56.0, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 2.0%, except where it is labeled as containing 100 mg of cefadroxil/mL after constitution, in which case the limit is NMT 3.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefadroxil RS
Cefadroxil Tablets
(Comment on this Monograph)id=m13955=Cefadroxil Tablets=Ca-Chl-Monos.pdf) DEFINITION Cefadroxil Tablets contain NLT 90.0% and NMT 120.0% of the labeled amount of C16H17N3O5S. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 2 mg/mL of USP Cefadroxil RS Sample solution: 2 mg/mL of cefadroxil from the powdered Tablets dissolved in water Chromatographic system (See Chromatography 621, System Suitability.) Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 20 L Pre-developing solvent solution: n-hexane and tetradecane (95:5) Developing solvent system: 0.1 M citric acid, 0.1 M dibasic sodium phosphate and a 1 in 15 solution of ninhydrin in acetone (60:40:15) Spray reagent: 1in500 solution of ninhydrin in dehydrated alcohol [NOTEProtect Spray reagent from light.] Analysis Samples: Standard solution and Sample solution Place the thin-layer chromatographic plate in a chamber containing the Pre-developing solvent solution and allow the solvent front to move the length of the plate. Remove the plate from the chamber and allow the solvent to evaporate. Apply the Sample solution and Standard solution to the plate, allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow to air-dry. Spray the
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USP 32
Meet the requirements NMT 8.0%
plate with the Spray reagent, dry for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Buffer solution: Dissolve 13.6 g of monobasic potassium phosphate in water to make 2000 mL of solution. Adjust with 10 N potassium hydroxide to a pH of 5.0. Mobile phase: Acetonitrile and Buffer solution (1:24) Standard solution: 1.06 mg/mL of USP Cefadroxil RS in Buffer solution [NOTEThis solution contains the equivalent of 1000 g/ mL of cefadroxil (C16H17N3O5S). Use this solution on the day prepared.] Sample solution: Weigh and finely powder NLT 10 Tablets. Transfer a portion of the powder, equivalent to 200 mg of cefadroxil, to a 200-mL volumetric flask, dilute with Buffer solution to volume, and stir by mechanical means for 5 min. Use this solution on the day prepared. Chromatographic system (See Chromatography 621, System suitability.) Mode: LC Detector: UV 230 nm Column: 4-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k : Between 2.03.5 Column efficiency: NLT 1800 theoretical plates Tailing factor: NMT 2.2 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O5S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) P F 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cefadroxil RS in the Standard solution (mg/mL) CU = nominal concentration of cefadroxil in the Sample solution (mg/mL) P = cefadroxil equivalent (g/mg) of USP Cefadroxil RS F = conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 30 min Sample solution: Sample per 711 Dissolution Analysis: Determine the amount of C16H17N3O5S dissolved from UV absorbances at the wavelength of maximum absorbance at about 263 nm of filtered portions of the Sample solution suitably diluted, if necessary, in comparison to a Standard solution having a known concentration of USP Cefadroxil RS. Tolerances: NLT 75% (Q) of the labeled amount of C16H17N3O5S rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The Tablets prepared using the hemihydrate form of cefadroxil are so labeled. USP REFERENCE STANDARDS 11 USP Cefadroxil RS
Cefamandole Nafate
(Comment on this Monograph)id=m13957=Cefamandole Nafate=Ca-Chl-Monos.pdf)
512.50 C19H17N6NaO6S2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[ [(formyloxy)phenylacetyl]amino]-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-, monosodium salt, [6R-[6,7 (R*)]]-; Sodium (6R,7R)-7-(R)-mandelamido-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylate formate (ester) [42540-40-9]. DEFINITION Cefamandole Nafate has a potency equivalent to NLT 810 g and NMT 1000 g of cefamandole (C18H18N6O5S2)/mg, calculated on the anhydrous basis. IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 10 mg/mL of USP Cefamandole Nafate RS in Developing solvent system [NOTEUse the solution promptly after preparation.] Sample solution: 10 mg/mL of Cefamandole Nafate in Developing solvent system [NOTEUse the solution promptly after preparation.] Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Ethyl acetate, acetone, glacial acetic acid, and water (5:2:1:1) Analysis Samples: Standard solution and Sample solution Place the plate in a suitable chromatographic chamber, previously equilibrated with Developing solvent system for NLT 30 min, and develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow to air-dry. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution
USP 32
ASSAY PROCEDURE Buffer solution: 3.6 mg/mL of anhydrous dibasic sodium phosphate, 39.4 mg/mL of citric acid monohydrate, and 70.8 mg/mL of potassium chloride Standard solution: Transfer 12 mg of USP Cefamandole Nafate RS to a 50-mL volumetric flask containing 4 mL of water. Immediately before use, add 30.0 mL of Buffer solution, and dilute with water to volume. Sample solution: Transfer 12 mg of Cefamandole Nafate to a 50-mL volumetric flask containing 4 mL of water. Immediately before use, add 30.0 mL of Buffer solution, and dilute with water to volume. Analysis: (See Polarography 801.) Samples: Standard solution and Sample solution Transfer a portion of the Sample solution to a suitable polarographic cell. Deaerate by bubbling scrubbed nitrogen through the solution for 5 min, and redirect the nitrogen flow to the surface outlet. Insert the dropping mercury electrode of a suitable polarograph capable of measuring a current of 0.5 microampere or appropriate current to maintain on-scale response, using an average capillary, and a drop rate of 1/s. Record the polarogram in the differential pulse mode from 0.3 volt to 1.05 volts, using a saturated calomel reference electrode and platinum wire counter electrode. Determine the peak height obtained, in microamperes, where the peak height is defined as the perpendicular distance from the extrapolated baseline to the highest point of the peak as compared to the full-scale current range. Similarly, determine the peak current of the Standard solution. Calculate the concentration, in g/mg, of C18H18N6O5S2 of the Cefamandole Nafate: Result = (iU/iS) (CS/CU) P = peak current, in microamperes, from the Sample solution = peak current, in microamperes, from the iS Standard solution = concentration of USP Cefamandole Nafate RS in CS the Standard solution (mg/mL) CU = concentration of Cefamandole Nafate taken to prepare the Sample solution (mg/mL) P = potency of cefamandole of USP Cefamandole Nafate RS (g/mg) Acceptance criteria: 810 g/mg1000 g/mg iU SPECIFIC TESTS PH 791: 3.57.0, in a solution containing 100 mg/mL WATER DETERMINATION, Method I 921: NMT 2.0% BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin Unit/mg of cefamandole, when the label states that Cefamandole Nafate is sterile or must be subjected to further processing during the preparation of injectable dosage forms STERILITY TESTS 71: Meets the requirements for Test for Sterility of the Product to be Examined, Membrane Filtration when the label states that Cefamandole Nafate is sterile ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefamandole Nafate RS USP Endotoxin RS
Official Monographs / Cefamandole 117
Cefamandole Nafate for Injection

(Comment on this Monograph)id=m13960=Cefamandole Nafate for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefamandole Nafate for Injection is a sterile mixture of Cefamandole Nafate and one or more suitable buffers. It has a potency equivalent to NLT 810 g and NMT 1000 g of cefamandole (C18H18N6O5S2)/mg, calculated on the anhydrous and sodium carbonate-free basis. It contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amount of cefamandole (C18H18N6O5S2). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 10 mg/mL of USP Cefamandole Nafate RS in Developing solvent system [NOTEUse the solution promptly after preparation.] Sample solution: 10 mg/mL of cefamandole nafate, from Cefamandole Nafate for Injection diluted with Developing solvent system. [NOTEUse the solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Ethyl acetate, acetone, glacial acetic acid, and water (5:2:1:1) Analysis Samples: Standard solution and Sample solution Analysis: Place the plate in a suitable chromatographic chamber, previously equilibrated with Developing solvent for NLT 30 min, and develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Buffer solution: 3.6 mg/mL of anhydrous dibasic sodium phosphate, 39.4 mg/mL of citric acid monohydrate, and 70.8 mg/mL of potassium chloride Standard solution: Transfer 12 mg of USP Cefamandole Nafate RS to a 50-mL volumetric flask containing 4 mL of water. Immediately before use, add 30.0 mL of Buffer solution, and dilute with water to volume. Sample stock solution A (where the article is represented as being in a single-dose container): Constitute a container of Cefamondole Nafate for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with water to obtain a solution containing 2 mg/mL of cefamandole. Sample solution A: Transfer 5.0 mL of Sample stock solution A to a 50-mL volumetric flask, add 30.0 mL of Buffer solution, and dilute with water to volume. Sample stock solution B (where the label states the quantity of cefamandole in a given volume of constituted solution): Constitute Cefamandole Nafate for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a volume of the constituted solution
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PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Analysis for content uniformity: Perform the Assay on individual containers using Sample solution A or Sample solution B, or both, as appropriate. SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Constituted Solutions, Injections 1. BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin Unit/mg of cefamandole STERILITY TESTS 71: It meets the requirements for Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791: 6.08.0, determined after 30 min in a solution containing 100 mg/mL PARTICULATE MATTER 788: Meets the requirements for small-volume injections WATER DETERMINATION, Method I 921: NMT 3.0% OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Cefamandole Nafate RS USP Endotoxin RS
with water to obtain a solution containing nominally 2 mg/mL of cefamandole. Sample solution B: Transfer 5.0 mL of Sample stock solution B to a 50-mL volumetric flask, add 30.0 mL of Buffer solution, and dilute with water to volume. Sample solution C: Transfer a quantity of Cefamondole Nafate for Injection, equivalent to 12 mg of Cefamandole Nafate, to a 50-mL volumetric flask containing 4 mL of water. Immediately before use, add 30.0 mL of Buffer solution, and dilute with water to volume. Determine the sodium carbonate content of a separate 1g portion of Cefamandole for Injection dissolved in 100 mL of water. Add methyl orange TS and titrate with 0.2 N sulfuric acid VS. Each mL of 0.2 N sulfuric acid is equivalent to 10.60 mg of Na2CO3. Analysis (See Polarography 801.) Samples: Standard solution and Sample solutions Transfer a portion of the Sample solution to a suitable polarographic cell. Deaerate by bubbling scrubbed nitrogen through the solution for 5 min, and redirect the nitrogen flow to the surface outlet. Insert the dropping mercury electrode of a suitable polarograph capable of measuring a current of 0.5 microampere or appropriate current to maintain on-scale response, using an average capillary, and a drop rate of 1/s. Record the polarogram in the differential pulse mode from 0.3 volt to 1.05 volts, using a saturated calomel reference electrode and platinum wire counter electrode. Determine the peak height obtained, in microamperes, where the peak height is defined as the perpendicular distance from the extrapolated baseline to the highest point of the peak as compared to the full-scale current range. Similarly, determine the peak current of the Standard solution. Calculate the percentage of C18H18N6O5S2 in the portion of constituted solution taken: Result = (iU/iS) (CS/CU) P F 100 = peak current, in microamperes, from the Sample solution iS = peak current, in microamperes, from the Standard solution = concentration of USP Cefamandole Nafate RS in CS the Standard solution (mg/mL) = nominal concentration of cefamandole in Sample CU solution A or in Sample solution B (mg/mL) P = potency of cefamandole of USP Cefamandole Nafate RS (g/mg) F = conversion factor, 0.001 mg/g Calculate the potency in g of C18H18N6O5S2/mg, of the Cefamandole Nafate for Injection taken: Result = (iU/iS) (CS/CU) P = peak current, in microamperes, from the Sample solution = peak current, in microamperes, from the iS Standard solution = concentration of USP Cefamandole Nafate RS in CS the Standard solution (mg/mL) = nominal concentration of cefamandole nafate in CU Sample solution C (mg/mL) P = potency of cefamandole of USP Cefamandole Nafate RS (g/mg) Acceptance criteria: 8101000 g/mg of C18H18N6O5S2, calculated on the anhydrous and sodium carbonate-free basis; 90.0%115.0% of the labeled amount of C18H18N6O5S2 iU iU
Cefazolin
(Comment on this Monograph)id=m13970=Cefazolin=Ca-ChlMonos.pdf)
454.51 C14H14N8O4S3 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[[(5methyl-1,3,4-thiadiazol-2-yl)thio]methyl]-8-oxo-7-[[1Htetrazol-1-yl)acetyl]amino]-, (6R-trans); (6R,7R)-3-[[(5-Methyl-1,3,4-thiadiazol-2-yl)thio]methyl]-8-oxo-7[2-(1H-tetrazol-1-yl)acetamido]-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylic acid [25953-19-9]. DEFINITION Cefazolin contains NLT 95.0% and NMT 103.0% of C14H14N8O4S3, calculated on the anhydrous basis. IDENTIFICATION The retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer A: 0.9 mg/mL of anhydrous dibasic sodium phosphate and 1.298 mg/mL of citric acid monohydrate Buffer B: 5.68 mg/mL of anhydrous dibasic sodium phosphate and 3.63 mg/mL of monobasic potassium phosphate Mobile phase: Acetonitrile and Buffer A (1:9) Internal standard solution: Dissolve 750 mg of salicylic acid in 10 mL of methanol and dilute to 100 mL with Buffer B.
USP 32
Standard stock solution: 1 mg/mL of USP Cefazolin RS in Buffer B Standard solution: Mix 5.0 mL of Standard stock solution and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Sample stock solution: 1 mg/mL of Cefazolin in Buffer B Sample solution: Mix 5.0 mL of Sample stock solution and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and cefazolin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between the analyte and internal standard peaks Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H14N8O4S3 in the portion of Cefazolin taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of Cefazolin to the internal standard from the Sample solution = peak response ratio of Cefazolin to the internal RS standard from the Standard solution = concentration of USP Cefazolin RS, calculated on CS the anhydrous basis, in the Standard solution (mg/mL) = concentration of Cefazolin in the Sample solution CU (mg/mL) Acceptance criteria: 95.0%103.0% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: RU
Official Monographs / Cefazolin 119

DEFINITION Cefazolin Sodium has a potency equivalent to NLT 89.1% and NMT 110.1% of cefazolin sodium (C14H13NaN8O4S3), calculated on the anhydrous basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: 20 g/mL in 0.1 M sodium bicarbonate B. The retention time of the major peak for cefazolin in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements ASSAY PROCEDURE Buffer A: 0.9 mg/mL of anhydrous dibasic sodium phosphate and 1.298 mg/mL of citric acid monohydrate in water Buffer B: 5.68 mg/mL of anhydrous dibasic sodium phosphate and 3.63 mg/mL of monobasic potassium phosphate in water Mobile phase: Acetonitrile and Buffer A (1:9) Pass through a membrane filter having a 10-m or finer porosity. Internal standard solution: 7.5 mg/mL of salicylic acid in methanol and Buffer B (1:9) Standard stock solution: 1 mg/mL of USP Cefazolin RS in Buffer B Standard solution: Mix 5.0 mL of Standard stock solution and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Sample stock solution: 1 mg/mL of Cefazolin Sodium in Buffer B Sample solution: Mix 5.0 mL of Sample stock solution and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and Cefazolin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between the analyte and the internal standard peaks Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C14H13NaN8O4S3 in the portion of Cefazolin Sodium taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 RU RS CS = peak response ratio of cefazolin to the internal standard from the Sample solution = peak response ratio of cefazolin to the internal standard from the Standard solution = concentration of USP Cefazolin RS, calculated on the anhydrous basis, in the Standard solution (mg/mL)
NMT 20 ppm NMT 2.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefazolin RS
Cefazolin Sodium
(Comment on this Monograph)id=m13972=Cefazolin Sodium=Ca-Chl-Monos.pdf) 476.49 C14H13N8NaO4S3 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[[(5methyl-1,3,4-thiadiazol-2-yl)thio]methyl]-8-oxo-7-[[(1Htetrazol-1-yl)acetyl]amino]-, monosodium salt (6R-trans); Monosodium (6R,7R)-3-[[(5-methyl-1,3,4-thiadiazol-2yl)thio]methyl]-8-oxo-7-[2-(1H-tetrazol-1-yl)acetamido]-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate [27164-46-1].
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Cefazolin / Official Monographs
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Sample solution: Mix 5.0 mL of Sample stock solution and 5.0 mL of Internal standard solution. Dilute to 100 mL with Buffer B. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for salicylic acid and cefazolin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between the analyte and internal standard peaks Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H14N8O4S3 in each mL of the Injection taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cefazolin to the internal standard from the Sample solution RS = peak response ratio of cefazolin to the internal standard from the Standard solution = concentration of USP Cefazolin RS, calculated on CS the anhydrous basis, in the Standard solution (mg/mL) CU = nominal concentration of cefazolin in the Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin Unit/mg of cefazolin STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 4.57.0 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements under Injections 1, Labeling. The label states that it is to be thawed just before use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cefazolin RS USP Endotoxin RS RU
= nominal concentration of Cefazolin Sodium in the Sample solution (mg/mL) = molecular weight of cefazolin sodium, 476.49 Mr1 = molecular weight of cefazolin, 454.51 Mr2 Acceptance criteria: 89.1%110.1% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 10 to 24 Sample solution: 55 mg/mL, in 0.1 M sodium bicarbonate PH 791: 4.06.0, in a solution containing 100 mg/mL of cefazolin WATER DETERMINATION, Method I 921: NMT 6.0% STERILITY TESTS 71: Where the label states that Cefazolin Sodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefazolin Sodium is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.15 USP Endotoxin Unit/mg of cefazolin. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefazolin RS USP Endotoxin RS
Cefazolin Injection
(Comment on this Monograph)id=m13974=Cefazolin Injection=Ca-Chl-Monos.pdf) DEFINITION Cefazolin Injection is a sterile solution of Cefazolin and Sodium Bicarbonate in a diluent containing one or more suitable tonicity-adjusting agents. It contains NLT 90.0% and NMT 115.0% of the labeled amount of cefazolin (C14H14N8O4S3). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer A: 0.9 mg/mL of anhydrous dibasic sodium phosphate and 1.298 mg/mL of citric acid monohydrate in water Buffer B: 5.68 mg/mL of anhydrous dibasic sodium phosphate and 3.63 mg/mL of monobasic potassium phosphate in water Mobile phase: Acetonitrile and Buffer A (1:9) Pass through a membrane filter having a 10-m or finer porosity. Internal standard solution: 7.5 mg/mL of salicylic acid in methanol and Buffer B (1:9) Standard stock solution: 1 mg/mL of USP Cefazolin RS in Buffer B Standard solution: Mix 5.0 mL of Standard stock solution and 5.0 mL of Internal standard solution. Dilute to 100 with Buffer B. Sample stock solution: Allow 1 container of Injection to thaw, and mix. Transfer a volume of the Injection, nominally equivalent to about 50 mg of cefazolin, to a 50-mL volumetric flask, dilute with Buffer B to volume, and mix.
Cefazolin for Injection

(Comment on this Monograph)id=m13978=Cefazolin for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefazolin for Injection contains an amount of Cefazolin Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of (C14H14N8O4S3).
USP 32
IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: 20 g/mL in 0.1 M sodium bicarbonate B. The retention time of the major peak for cefazolin from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements ASSAY PROCEDURE Buffer A: 0.9 mg/mL of anhydrous dibasic sodium phosphate and 1.298 mg/mL of citric acid monohydrate in water Buffer B: 5.68 mg/mL of anhydrous dibasic sodium phosphate and 3.63 mg/mL of monobasic potassium phosphate in water Mobile phase: Acetonitrile and Buffer A (1:9) Pass through a membrane filter having a 10-m or finer porosity. Internal standard solution: 7.5 mg/mL of salicylic acid in methanol and Buffer B (1:9) Standard stock solution: 1 mg/mL of USP Cefazolin RS in Buffer B Standard solution: Mix 5 mL of Standard stock solution and 5 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Sample stock solution A (where it is packaged for dispensing and is represented as being in a single-dose container): Constitute Cefazolin for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with Buffer B to obtain a solution containing nominally 1 mg/mL of cefazolin. Sample solution A: Mix 5.0 mL of Sample stock solution A and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Sample stock solution B: (where the label states the quantity of cefazolin in a given volume of constituted solution): Constitute Cefazolin for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a volume of the constituted solution with Buffer B to obtain a solution containing nominally 1 mg/mL of cefazolin. Sample solution B: Mix 5.0 mL of Sample stock solution A and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times are about 0.7 for salicylic acid and 1.0 for Cefazolin.] Suitability requirements Resolution: NLT 4.0 between the analyte and internal standard peaks Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution A, Sample solution B, and Standard solution Calculate the percentage of C14H14N8O4S3 in the container, and in the volume of constituted solution: Result = (RU/RS) (CS/CU) 100 RU
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= peak response ratio of cefazolin to the internal standard of the Sample solution = peak response ratio of cefazolin to the internal RS standard of the Standard solution = concentration of USP Cefazolin RS, calculated on CS the anhydrous basis, in the Standard solution (mg/mL) = nominal concentration of cefazolin in the Sample CU solution (mg/mL) [NOTEWhere the test for Uniformity of Dosage Units has been performed using the Analysis for content uniformity, use the average of these determinations as the Assay value.] Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Analysis for content uniformity: Perform the Assay on individual containers using Sample solution A or Sample solution B, or both, as appropriate. SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. OPTICAL ROTATION, Specific Rotation 781S: 10 to 24 Sample solution: 55 mg/mL, in 0.1 M sodium bicarbonate BACTERIAL ENDOTOXINS TEST 85: NMT 0.15 USP Endotoxin Unit/mg of cefazolin STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791: 4.06.0, in a solution containing 100 mg/mL of cefazolin WATER DETERMINATION, Method I 921: NMT 6.0% PARTICULATE MATTER IN INJECTION 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: It meets the requirements for Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. USP REFERENCE STANDARDS 11 USP Cefazolin RS USP Endotoxin RS
Cefazolin Ophthalmic Solution

(Comment on this Monograph)id=m13980=Cefazolin Ophthalmic Solution=Ca-Chl-Monos.pdf) DEFINITION Cefazolin Ophthalmic Solution contains an amount of Cefazolin Sodium equivalent to NLT 29.7 mg and NMT 36.3 mg of cefazolin (C14H14N8O4S3) in 10.0 mL of Ophthalmic Solution. Use Cefazolin Sodium or Cefazolin for Injection that contains the designated amount of cefazolin, and prepare the Ophthalmic Solution as follows (see Pharmaceutical CompoundingNonsterile Preparations 795):
Cefazolin Sodium Thimerosal Sodium Chloride Injection (0.9%) To make 35 mg 0.2 mg A sufficient quantity 10.0 mL
Dissolve quantities of Cefazolin Sodium and Thimerosal in Sodium Chloride Injection (0.9%), and dilute quantitatively and stepwise if necessary, with Sodium Chloride Injection (0.9%) to obtain a solution containing, in each mL, 3.5 mg of Cefazolin Sodium and 0.02 mg of Thimerosal. Filter a 10.0-mL
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F
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= correction factor (to convert mg/mL to mg/10 mL), 10 Acceptance criteria: 29.736.3 mg SPECIFIC TESTS STERILITY: See Pharmaceutical CompoundingNonsterile Preparations 795, Sterility PH 791: 4.56.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, sterile ophthalmic containers. Store in a refrigerator. LABELING: Label it to state that it is intended for use in the eye, and is not to be used if a precipitate is present. BEYOND-USE DATE: 5 days after the date on which it was compounded USP REFERENCE STANDARDS 11 USP Cefazolin RS
portion of the resulting solution to produce a clear and sterile Ophthalmic Solution. If Cefazolin for Injection is used, prepare the Ophthalmic Solution as follows. Dissolve an accurately weighed quantity of Thimerosal in Sodium Chloride Injection (0.9%), and dilute quantitatively and stepwise if necessary, with Sodium Chloride Injection (0.9%) to obtain a solution containing 0.3 mg of Thimerosal/mL. Add 9.8 mL of the resulting solution to a vial of Cefazolin for Injection, containing 500 mg of cefazolin, and mix to obtain a stock solution. Transfer 3.3 mL of the stock solution to a 50-mL volumetric flask, dilute with Sodium Chloride Injection (0.9%) to volume, and mix. Filter a 10.0-mL portion of the resulting solution to produce a clear and sterile Ophthalmic Solution. ASSAY PROCEDURE Buffer A: 0.9 mg/mL of anhydrous dibasic sodium phosphate and 1.298 mg/mL of citric acid monohydrate in water Buffer B: 5.68 mg/mL of anhydrous dibasic sodium phosphate and 3.63 mg/mL of monobasic potassium phosphate in water Solution C: Acetonitrile and Buffer A (1:9) Pass the resulting solution through a filter having a 5-m or finer porosity. Solution D: Acetonitrile and Buffer A (4:1) Pass the resulting solution through a filter having a 5-m or finer porosity. Mobile phase: See the gradient table below.
Time (min) 0 15 25 Solution C (%) 100 0 100 Solution D (%) 0 100 0
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Cefdinir
(Comment on this Monograph)id=m13982=Cefdinir=Ca-ChlMonos.pdf)
Standard solution: 0.32 mg/mL of USP Cefazolin RS in Buffer B Maintain at 4 prior to injection. Sample solution: Transfer 1.0 mL of Ophthalmic Solution to a 10-mL low-actinic volumetric flask, dilute with Buffer B to volume, and mix. Maintain at 4 prior to injection. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 273 nm Column: 3.9-mm 30-cm; 10-m packing L1 Temperature: 25 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the quantity, in mg, of C14H14N8O4S3 in the portion of Ophthalmic Solution taken: Result = (rU/rS) CS D F rU rS CS D = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cefazolin RS in the Standard solution (mg/mL) = dilution factor used in the preparation of the Sample solution, 10
C14H13N5O5S2 395.41 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2amino-4-thiazolyl)(hydroxyimino)acetyl]amino]-3-ethenyl-8oxo-, [6R-[6, 7(Z)]]-; (-)-(6R, 7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-8-oxo-3vinyl-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid, 72-(Z)-oxime [91832-40-5]. DEFINITION Cefdinir contains NLT 960 g/mg and NMT 1020 g/mg of C14H13N5O5S2, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 14.2 mg/mL anhydrous dibasic sodium phosphate Solution B: 13.6 mg/mL monobasic potassium phosphate Buffer solution: Combine appropriate amounts of Solution A and Solution B (about 2:1) to obtain a pH 7.0 solution. Solution C: Dilute tetramethylammonium hydroxide (10%) with water to obtain a 1% solution. Adjust with dilute phosphoric acid (1 in 10) to a pH of 5.5. Solution D: 37.2 mg/mL edetate disodium Mobile phase: Acetonitrile, methanol, Solution C, and Solution D (60:40:900:0.4) System suitability solution: 0.2 mg /mL of USP Cefdinir RS and 0.5 mg/mL of USP Cefdinir Related Compound A RS, Buffer solution
USP 32
Standard solution: 0.2 mg/mL of USP Cefdinir RS, Buffer solution Sample solution: 0.2 mg/mL of Cefdinir, Buffer solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5 m, packing L1 Temperature: 40 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution and System suitability solution [NOTEUSP Cefdinir Related Compound A RS should produce four peaks.] Tailing factor: NMT 1.5 for cefdinir, System suitability solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Sample: Standard solution and Sample solution Calculate the quantity (g/mg) of C14H13N5O5S2 in the portion of Cefdinir taken: Result = (rU/rS) (CS/CU) P rU = peak response from the Sample solution = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU P = purity of USP Cefdinir RS (g/mg) Acceptance criteria: 9601020 g/mg IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: 10 ppm Organic Impurities PROCEDURE Solution A, Solution B, Buffer solution, Solution C, and Solution D: Prepare as directed in the Assay. Solution E: To 1000 mL of Solution C, add 0.4 mL of Solution D. Solution F: Acetonitrile, methanol, Solution C, and Solution D (300:200:500:0.4) System suitability solution A: 15 g/mL of cefdinir, from the Sample solution, diluted with Solution C System suitability solution B: 1.5 g /mL of cefdinir, from System suitability solution A, diluted with Solution C System suitability solution C: Transfer about 30 mg of USP Cefdinir RS and 2 mg of USP Cefdinir Related Compound A RS to a 20-mL volumetric flask, dissolve in 3 mL of Buffer solution, and dilute with Solution C to volume. Sample stock solution: 10 mg/mL of Cefdinir in Buffer solution Sample solution: 1.5 mg/mL of Cefdinir from the Sample stock solution, in Solution C [NOTEPrepare fresh immediately before use.] Mobile phase: See the gradient table below.
Time (min) 0 2 22 Solution E (%) 95 95 75 Solution F (%) 5 5 25 Time (min) 32 37 38 58
Official Monographs / Cefdinir 123

Solution E (%) 50 50 95 95 Solution F (%) 50 50 5 5
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5 micron packing L1 Temperature: 40 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: System suitability solution A, System suitability solution B, and System suitability solution C [NOTEUSP Cefdinir Related Compound A RS should produce four peaks.] [NOTEThe relative retention time of the third peak from USP Cefdinir Related compound A RS is NLT 1.1, relative to the cefdinir peak, System suitability solution C.] Suitability requirements Linearity: The response of cefdinir in System suitability solution B is between 7% and 13% of that from System suitability solution A. Column efficiency: NLT 7000 theoretical plates for cefdinir, System suitability solution C Tailing factor: NMT 3.0 for cefdinir, System suitability solution C Relative standard deviation: NMT 2.0% for cefdinir, System suitability solution C Analysis Sample: Sample solution [NOTERecord the chromatogram for NLT 40 min.] Calculate the percentage of each impurity in the portion of Cefdinir taken: Result = (rU/rS) 100 rU r = peak response of each impurity from the Sample solution = sum of all the peak responses from the Sample solution
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Cefdinir / Official Monographs
USP 32
IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Buffer: Prepare as directed in the Assay. Blank: Use the Buffer. Standard solution: 10 g/mL of USP Cefdinir RS in Buffer Sample solution: Equivalent to 10 g/mL of Cefdinir in Buffer Filter before use. Cell size: 1 cm Acceptance criteria: Absorptivities of the Sample solution maxima and minima occur at the same wavelengths as those in the Standard solution. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: 10.65 mg/mL of dibasic sodium phosphate and 3.40 mg/mL of monobasic potassium phosphate Adjust to pH 7.0 0.05 with phosphoric acid or sodium hydroxide before final dilution. Solution A: 7.0 mg/mL citric acid monohydrate Adjust to pH 2.0 0.05 with phosphoric acid. Mobile phase: Methanol, tetrahydrofuran, and Solution A (111:28:1000) System suitability solution: 50 g/mL of cefdinir and 175 g/mL of m-hydroxybenzoic acid in Buffer Standard solution: 50 g/mL of USP Cefdinir RS in Buffer Sample solution: Equivalent to 50 g/mL of Cefdinir, from Capsule contents in the Buffer Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; 4 m, packing L1 Flow rate: 1.4 mL/min Injection size: 15 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: NLT 3.0 between cefdinir and mhydroxybenzoic acid, System suitability solution Tailing factor: NMT 2.0 for cefdinir, System suitability solution Relative standard deviation: NMT 1.0% for cefdinir, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of Cefdinir in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response for cefdinir from the Sample solution = peak response for cefdinir from the Standard solution = concentration of the Standard solution (mg/mL) = nominal concentration of cefdinir in the Sample solution (mg/mL)
Impurity Table 1 Relative Retention Time 0.10 0.12 0.74 0.85, 0.93, 1.11, 1.14 1.22 1.36 1.51 1.61, 1.64 Acceptance Criteria, NMT (%) 0.5 0.5 0.7 0.7 (sum of 4) 0.5 0.5 0.7 0.5 (sum of 2) 0.2
Name Impurity Aa Impurity Bb Impurity Cc Cefdinir related compound Ad (4 peaks) Impurity Ee Impurity Ff Impurity Gg Impurity Hh (2 peaks) Any other individual, unidentified impurity
a
1N-[(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)acetyl]glycine. b(Z)-2-(2-Aminothiazol-4-yl)-N-(2,2-dihydroxyethyl)-2(hydroxyimino)acetamide. c(6R,7R)-7-[(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)acetamido]-3methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. d2(R)-2-[(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)acetamido)]-2[(2RS,5RS)-5-methyl-7-oxo-2,4,5,7-tetrahydro-1H-furo[3,4-d][1,3]thiazin-2yl]acetic acid. e(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)-N-{(3RS,5aR,6R)-3methyl-1,7-dioxo-1,3,4,5a,6,7-hexahydroazeto[2,1-b]furo[3,4-d] [1,3]thiazin-6-yl}acetamide. f(6R,7R)-7-(4-hydroxyisoxazole-3-carboxamido)-8-oxo-3-vinyl-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. g(6R,7R)-7-[(E)-2-(2-Aminothiazol-4-yl-)-2-(hydroxyimino)acetamido]-8oxo-3-vinyl-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. h(Z)-2-(2-Aminothiazol-4-yl)-2-(hydroxyimino)-N-{[(2RS,5RS)-5-methyl-7oxo-2,4,5,7-tetrahydro-1H-furo[3,4-d][1,3]thiazin-2-yl]methyl}acetamide.
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 61 to 67 at 20 Sample solution: 10 mg/mL in the Buffer Solution as in the Assay WATER DETERMINATION, Method I 921: NMT 2.0% for anhydrous; 4.0%8.0% for monohydrate, using formamide and methanol (2:1) as the solvent ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Cefdinir RS USP Cefdinir Related Compound A RSvUSP32
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Cefdinir Capsules
(Comment on this Monograph)id=m2183=Cefdinir Capsules=Ca-Chl-Monos.pdf) DEFINITION Cefdinir Capsules contains NLT 90.0% and NMT 110.0% of the labeled amount of Cefdinir (C14H13N5O5S2).
USP 32

flask. Dissolve in 30 mL of Buffer, and dilute with Diluent to volume to obtain a solution having a known concentration of about 1.5 mg/mL of cefdinir. Mobile phase: See the gradient table below.
Time (min) 0 2 22 32 37 38 58 Solution E (%) 95 95 75 50 50 95 95 Solution F (%) 5 5 25 50 50 5 5
PERFORMANCE TESTS DISSOLUTION 711 Medium: 50 mM phosphate buffer pH 6.8; 900 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 290 nm Standard solution: 0.33 mg/mL USP Cefdinir RS in Medium Sample solution: Sample per Dissolution 711 Filter each sample through a suitable 0.45 m filter. Dilute with Medium to a concentration of about 0.33 mg/mL of cefdinir. Blank: Dissolve one empty capsule in 100 mL of Medium, and dilute to 900 mL; filter if necessary. Analysis: Determine the percentage of C14H13N5O5S2 dissolved: Result = (AU/AS) CS D (V/L) 100 AU = absorbance of the Sample solution = absorbance of the Standard solution AS = concentration of the Standard solution (mg/mL) CS D = dilution factor of the Sample solution (mL/mL) V = volume of Medium (mL), 900 L = Label claim (mg) Tolerances: NLT 80% (Q) of the labeled amount of C14H13N5O5S2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Solution A: 14.2 mg/mL anhydrous dibasic sodium phosphate Solution B: 13.6 mg/mL monobasic potassium phosphate Buffer: Combine appropriate amounts of Solution A and Solution B (about 2:1) to obtain a pH 7.0 0.1 solution. Diluent: Dilute tetramethylammonium hydroxide (10%) with water to obtain a 1% solution. Adjust with dilute phosphoric acid (1 in 10) to a pH of 5.5 0.1. Solution D: 37.2 mg/mL edetate disodium Solution E: To 1000 mL of Diluent, add 0.4 mL of Solution D. Solution F: Acetonitrile, methanol, Diluent, and Solution D (150:100:250:0.2) Standard stock solution: 750 mg/mL of USP Cefdinir RS, Buffer Standard solution: 15 g/mL of USP Cefdinir RS, from the Standard stock solution in Diluent System suitability stock solution 1: 40 g/mL of USP Cefdinir Related Compound A RS in Diluent System suitability stock solution 2: 40 g/mL of USP Cefdinir Related Compound B RS in Diluent System suitability solution: Transfer 37.5 mg of USP Cefdinir RS to a 25-mL volumetric flask. Add about 10 mL of Buffer. Add 5.0 mL of each of System suitability stock solution 1 and System suitability stock solution 2, and dilute with Diluent to volume. Sample solution: Transfer an equivalent to 300 mg of Cefdinir, from Capsule contents, into a 200-mL volumetric
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: Column: 4.6-mm 15-m; packing L1 Column temperature: 40 0.5 Sample solution temperature: 4 3 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: NLT 1.5, between cefdinir and the third peak of the USP Cefdinir Related Compound A RS, System suitability solution Tailing factor: NMT 1.5 for cefdinir related compound B, System suitability solution Relative standard deviation: NMT 2.0%, for cefdinir peak response, Standard solution Analysis Sample: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100/RRF rU = peak response from the Sample solution = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU RRF = relative response factor (see Impurity Table 1.) Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cefdinir RS USP Cefdinir Related Compound A RS USP Cefdinir Related Compound B RSvUSP32
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m filter, and transfer 5.0 mL of the filtrate to a 10-mL volumetric flask, and dilute with methanol to volume. Application volume: 10 L Developing solvent system: Methanol and water (4:1) Visualization: Shortwave UV Analysis Sample: Standard solution and Sample solution Develop the chromatogram until the solvent front has moved about 15 cm. Remove the plate from the developing chamber, and allow the solvent to evaporate. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: 10.65 mg/mL of anhydrous dibasic sodium phosphate and 3.40 mg/mL of monobasic potassium phosphate in water Adjust to pH 7.0 0.05 with phosphoric acid or sodium hydroxide before final dilution.
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Cefdinir for Oral Suspension
(Comment on this Monograph)id=m2184=Cefdinir for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cefdinir for Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of C14H13N5O5S2. It may contain one or more suitable buffers, flavors, preservatives, stabilizing agents, sweeteners, and suspending agents. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel, preconditioned with n-hexane and tetradecane (95:5) Buffer: Prepare as directed in the Assay. Standard solution: 600 g/mL of USP Cefdinir RS in methanol and Buffer (3:1) Sample solution: Transfer an equivalent to 125 mg of Cefdinir from reconstituted Suspension, in a 100-mL volumetric flask, add 50 mL of Buffer and dilute with methanol to volume. Pass a portion through a suitable 0.45-
Impurity Table 1 Relative Retention Time 0.10 0.13 0.36 0.46 0.77 0.75 0.85 0.94 1.11 1.14 1.18 1.23 1.28 1.37 1.44 1.49 1.51 1.62 1.64 1.82 Relative Response Factor 1.1 1.1 1.0 1.5 1.0 1.0 1.5 1.5 1.5 1.5 1.1 1.2 1.1 1.4 1.0 1.0 1.1 1.3 1.3 1.0 1.0 Limit of Quantification (% Cefdinir) 0.1 0.1 0.05 0.05 0.05 0.05 0.1 0.1 0.1 0.1 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Acceptance Criteria, NMT (%) 0.5 0.5 0.2 0.7 0.3 0.7 2.5 0.2 1.0 0.2 0.5 0.5 0.2 0.7 1.0 0.2 0.2 1.0
Name Impurity VIII Impurity IV Impurity XIV Impurity V Impurity B Impurity XI Cefdinir Related Compound A (lactam ring cleavage lactones-a)a Cefdinir Related Compound A (lactam ring cleavage lactones-b)a Cefdinir Related Compound A (lactam ring cleavage lactones-c)a Cefdinir Related Compound A (lactam ring cleavage lactones-d)a Impurity VI Impurity I Cefdinir Related Compound Ba Impurity XIII Impurity Ec Impurity XV Impurity VII Impurity IIIab Impurity IIIbb Impurity Dc Individual unidentified impurities Total unidentified impuritiesd
a
RS II is a mixture of 4 isomers designated as RS IIa, RS IIb, RS IIc, and RS IId. The sum of all values is reported and the total limit for all 4 isomers combined is 2.5%. bRS III is a mixture of 2 isomers designated as RS IIIa and RS IIIb. The sum of both values is reported and the total limit for both isomers combined is 1.0%. cImpurity B, Impurity D, and Impurity E are unidentified impurities. dThe total unidentified impurities limit includes the % total of unidentified impurities B, D, and E and any other individual unidentified impurities.
USP 32
Solution A: 7.0 mg/mL citric acid monohydrate Adjust to pH 2.0 0.05 with phosphoric acid. Mobile phase: Methanol, tetrahydrofuran, and Solution A (111:28:1000) System suitability solution: 50 g/mL of cefdinir and 175 g/mL of m-hydroxybenzoic acid in Buffer Standard solution: 50 g/mL of USP Cefdinir RS in Buffer Sample solution: Equivalent to 50 g/mL of Cefdinir, from constituted Suspension in Buffer Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; 4 m, packing L1 Flow rate: 1.4 mL/min Injection size: 15 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: NLT 3.0 between cefdinir and mhydroxybenzoic acid, System suitability solution Tailing factor: NMT 2.0 for cefdinir, System suitability solution Relative standard deviation: NMT 1.0% for cefdinir, Standard solution Analysis Sample: Standard solution and Sample solution Calculate the percentage of Cefdinir in the portion of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response for cefdinir from the Sample solution = peak response for cefdinir from the Standard rS solution = concentration of the Standard solution (mg/mL) CS = nominal concentration of cefdinir in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 50 mM phosphate buffer pH 6.8; 900 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 290 nm Sample solution: Dilute a portion of each filtered sample with Medium as necessary to obtain a solution having a concentration of about 0.14 mg per mL of cefdinir. Standard solution: 0.14 mg/mL USP Cefdinir RS in Medium Blank: Medium Analysis: Transfer 5 mL, by weight, of the reconstituted Oral Suspension into the vessel. After the appropriate time, withdraw a portion of the solution under test and pass through a suitable 0.45-m filter. Determine the percentage of C14H13N5O5S2 dissolved: Result = (AU/AS) ([CS d D V]/W L) 100 AU AS CS d D V W L = = = = = = = = absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (mg/mL) density of the Oral Suspension (mg/mL) dilution factor of the Sample solution (mL/mL) volume of Medium (mL), 900 weight of Oral Suspension taken (mg) Label claim (mg) rU

UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements DELIVERABLE VOLUME 698 (for Oral Suspension packaged in multiple-unit containers): Meets the requirements IMPURITIES Organic Impurities PROCEDURE Solution A: 14.2 mg/mL anhydrous dibasic sodium phosphate Solution B: 13.6 mg/mL monobasic potassium phosphate Buffer: Combine appropriate amounts of Solution A and Solution B (about 2:1) to obtain a pH 7.0 0.1 solution. Diluent: Dilute tetramethylammonium hydroxide (10%) with water to obtain a 1% solution. Adjust with dilute phosphoric acid (1 in 10) to a pH of 5.5 0.1. Solution D: 37.2 mg/mL edetate disodium Solution E: To 1000 mL of Diluent, add 0.4 mL of Solution D. Solution F: Acetonitrile, methanol, Diluent, and Solution D (150:100:250:0.2) Standard stock solution: 750 g/mL of USP Cefdinir RS in Buffer Standard solution: 15 g/mL of USP Cefdinir RS, from the Standard stock solution in Diluent System suitability stock solution 1: 40 g/mL of USP Cefdinir Related Compound A RS in Diluent System suitability stock solution 2: 40 g/mL of USP Cefdinir Related Compound B RS in Buffer System suitability solution: Transfer 37.5 mg of USP Cefdinir RS to a 25-mL volumetric flask. Add about 10 mL of Buffer. Add 5.0 mL of each of System suitability stock solution 1 and System suitability stock solution 2, and dilute with Diluent to volume. Sample solution: Transfer an equivalent to 150 mg of Cefdinir, from constituted Oral Suspension, into a 100-mL volumetric flask. Dissolve in 30 mL of Buffer, and dilute with Diluent to volume. Mobile phase: See the gradient table below.
Time (min) 0 2 22 32 37 38 58 Solution E (%) 95 95 75 50 50 95 95 Solution F (%) 5 5 25 50 50 5 5
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5 m packing L1 Column temperature: 40 0.5 Sample solution temperature: 4 3 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: NLT 1.5, between cefdinir and the third peak of the USP Cefdinir Related Compound A RS, System suitability solution
128

USP 32
Tailing factor: NMT 1.5 for cefdinir related compound B, System suitability solution Relative standard deviation: NMT 2.0% for cefdinir peak response, Standard solution Analysis Sample: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100/RRF rU rS CS CU RRF = peak response of impurity from the Sample solution = peak response from the Standard solution = concentration of the Standard solution (mg/mL) = nominal concentration of cefdinir in the Sample solution (mg/mL) = relative response factor (see Impurity Table 1)
SPECIFIC TESTS PH 791: 3.54.5 LOSS ON DRYING 731: Dry about 1 g over phosphorous pentoxide in a vacuum not exceeding 5 mm of mercury at 70 for 44.5 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight light-resistant containers, and store at controlled room temperature. LABELING: The label specifies the directions for the constitution of the powder and states the equivalent amount of C14H13N5O5S2 in a given volume of Oral Suspension after constitution.
Impurity Table 1 Relative Retention Time 0.10 0.13 0.36 0.46 0.77 0.75 0.85 Relative Response Factor 1.1 1.1 1.0 1.5 1.0 1.0 1.5 Limit of Quantification (% Cefdinir) 0.1 0.1 0.05 0.05 0.05 0.05 0.1 Acceptance Criteria, NMT (%) 0.5 0.6 0.2 0.3 0.2 0.7 3.3
Name Impurity VIII Impurity IV Impurity XIV Impurity V Impurity Bc Impurity XI Cefdinir Related Compound A (lactam ring cleavage lactonesa)a Cefdinir Related Compound A (lactam ring cleavage lactonesb)a Cefdinir Related Compound A (lactam ring cleavage lactonesc)a Cefdinir Related Compound A (lactam ring cleavage lactonesd)a Impurity VI Impurity I Cefdinir Related Compound B Impurity XIII Impurity Ec Impurity XV Impurity VII Impurity IIIab Impurity IIIbb Impurity Dc Individual unidentified impurities Total unspecified impuritiesd
a
0.94
1.5
0.1
1.11
1.5
0.1
1.14
1.5
0.1
1.18 1.23 1.28 1.37 1.44 1.49 1.51 1.62 1.64 1.82
1.1 1.2 1.1 1.4 1.0 1.0 1.1 1.3 1.3 1.0 1.0
0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05
0.2 0.8 0.2 0.5 0.2 0.2 1.2 1.1 0.2 0.2 0.9
Cefdinir related compound A is a mixture of 4 isomers designated as lactam ring cleavage lactones a, b, c, and d. The sum of all values is reported, and the total limit for all 4 isomers combined is 3.3%. bImpurity III is a mixture of 2 isomers designated as Impurity IIIa and Impurity IIIb. The sum of both values is reported, and the total limit for both isomers combined is 1.5%. cImpurity B, Impurity D, and Impurity E are unidentified impurities. dThe total unidentified impurities limit includes the % total of unidentified impurities B, D, and E and any other unidentified impurities detected.
USP 32
USP REFERENCE STANDARDS 11 USP Cefdinir RS USP Cefdinir Related Compound A RS USP Cefdinir Related Compound B RSvUSP32 CS
Official Monographs / Cefepime 129

= concentration of USP Cefepime Hydrochloride RS in the Standard solution (mg/mL) = concentration of Cefepime Hydrochloride in the CU Sample solution (mg/mL) P = content of cefepime in USP Cefepime Hydrochloride RS (g/mg) Acceptance criteria: 825911 g/mg IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1 : LIMIT OF N-METHYLPYRROLIDINE Mobile phase: Acetonitrile and 0.01 N nitric acid (1:100) Column rinse solution: Transfer 5.0 mL of nitric acid to a 1-L volumetric flask. Dilute with water to volume, and mix. Transfer this solution to an appropriate flask, add 1 L of acetonitrile, and mix. Standard stock solution: Weigh 0.16 mL of Nmethylpyrrolidine. Transfer to a 100-mL volumetric flask and dilute with water to volume. Standard solution: 4 mL of Standard stock solution diluted to 100 mL with 0.01 N nitric acid [NOTEThis solution contains 0.05 mg/mL of Nmethylpyrrolidine.] Sample solution: 10 mg/mL of Cefepime Hydrochloride in 0.01 N nitric acid [NOTEThis solution may be kept up to 6 h if maintained at 5; otherwise, use this solution within 30 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Conductivity [NOTEThe typical background conductance is 3500 S.] Column: 4.6-mm 5-cm; 5-m packing L52 Guard column: 4.4-mm 5-cm; packing L17, placed between the pump and the injector Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Retention time: NLT 8 min for N-methylpyrrolidine Relative standard deviation: NMT 5.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of N-methylpyrrolidine in the portion of Cefepime Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response of N-methylpyrrolidine from the Sample solution = peak response of N-methylpyrrolidine from the rs Sample solution = concentration of N-methylpyrrolidine in the CS Standard solution (mg/mL) = concentration of Cefepime Hydrochloride in the CU Sample solution (mg/mL) Acceptance criteria: NMT 0.3% [NOTECefepime from the Sample solution elutes as a broad peak at 55 min. To minimize the equilibration time at the start of the next day, it is recommended that the detector be turned on the night before and that the Mobile phase be pumped through the system overnight at a flow rate of 0.2 mL/min. After every Sample solution injection, it is recommended that the chromatograph be flushed with Column rinse solution for 30 min at a flow rate of 1 mL/min to remove cefepime from the column, rU
Cefepime Hydrochloride
(Comment on this Monograph)id=m13984=Cefepime Hydrochloride=Ca-Chl-Monos.pdf)
571.50 C19H25ClN6O5S2 HCl H2O Pyrrolidinium, 1-[[7-[[(2-amino-4thiazolyl)(methoxyimino)acetyl]amino]-2-carboxy-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-1-methyl-, chloride, monohydrochloride, monohydrate, [6R-[6,7(Z)]]-; 1-[[(6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-2carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3yl]methyl]-1-methylpyrrolidinium chloride, 72-(Z)-(Omethyloxime), monohydrochloride, monohydrate [123171-59-5]. DEFINITION Cefepime Hydrochloride contains the equivalent of NLT 825 g and NMT 911 g of cefepime (C19H24N6O5S2)/mg, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197M Sample: Proceed as directed in the chapter, but do not dry. ASSAY PROCEDURE Solution A: Dissolve 5.76 g of sodium 1-pentanesulfonate in 2000 mL of water, adjust with glacial acetic acid to a pH of 3.4, and then with potassium hydroxide TS to a pH of 4.0. Mobile phase: Acetonitrile and Solution A (3:47) Standard solution: 1.4 mg/mL of USP Cefepime Hydrochloride RS in Mobile phase Sample solution: 1.4 mg/mL of Cefepime Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.7 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C19H24N6O5S2 in each mg of Cefepime Hydrochloride taken: Result = (rU/rS) (CS/CU) P rU rS = peak response from the Sample solution = peak response from the Standard solution
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and that the system then be switched back to Mobile phase at a flow rate of 1 mL/min for reequilibration.]
USP 32
SPECIFIC TESTS BACTERIAL ENDOTOXINS TESTS 85: Where the label states that Cefepime Hydrochloride is sterile or that it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.04 USP Endotoxin Unit/mg of cefepime hydrochloride. STERILITY TESTS 71: Where the label states that Cefepime Hydrochloride is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. CRYSTALLINITY 695: Meets the requirements WATER DETERMINATION, Method I 921: 3.0%4.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefepime Hydrochloride RS USP Cefepime Hydrochloride System Suitability RS USP Endotoxin RS
PROCEDURE 2 Solution A: 0.68mg/mL of monobasic potassium phosphate Solution B: Acetonitrile and Solution A (1:9) Adjust with potassium hydroxide or phosphoric acid to a pH of 5.0. Solution C: Acetonitrile and Solution A (1:1) Adjust with potassium hydroxide or phosphoric acid to a pH of 5.0. Mobile phase: See the gradient table below.
System suitability solution: 1.4 mg/mL of USP Cefepime Hydrochloride System Suitability RS in Solution B Sample solution: 1.4 mg/mL of Cefepime Hydrochloride in Solution B [NOTEInject this solution immediately, or store in a refrigerator, and inject within 12 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Sample solution Suitability requirements Resolution: NLT 5 between cefepime and cefepime related compound A, and NLT 10 between cefepime related compound A and cefepime related compound B, System suitability solution Capacity factor, k: NLT 0.6, Sample solution Column efficiency: NLT 4000 theoretical plates, Sample solution Tailing factor: NMT 1.5, Sample solution Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Cefepime Hydrochloride taken: Result = (rU/rT) 100 = peak response for each impurity rU = sum of all the peak responses rT Acceptance criteria: See the Impurity Table 1.
Impurity Table 1 Relative Retention Time (%) 1.0 2.7 4.3 Relative Response Factor Acceptance Criteria, NMT (%) 0.3 0.1 0.1
Cefepime for Injection

(Comment on this Monograph)id=m13987=Cefepime for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefepime for Injection is a sterile mixture of Cefepime Hydrochloride and Arginine. It contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amount of cefepime (C19H24N6O5S2). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 20 mg/mL of arginine Sample solution: 40 mg/mL of Cefepime for Injection Developing solvent system: n-Propyl alcohol, ammonium hydroxide, and water (7:4:5) Analysis Samples: Sample solution and Standard solution Proceed as directed in Chromatography 621, Thin-Layer Chromatography, except to spray the plate with ninhydrin TS. Acceptance criteria: Arginine appears as a dark red spot. The intensity and the RF value of the spot from the Sample solution correspond to those from the Standard solution. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 2.88 mg/mL of sodium 1-pentanesulfonate Adjust with glacial acetic acid to a pH of 3.4, and then with potassium hydroxide TS to a pH of 4.0. Mobile phase: Acetonitrile and Solution A (3:47) Standard solution: 1.4 mg/mL of USP Cefepime Hydrochloride RS in Mobile phase Sample solution: 1 mg/mL of cefepime from one container of Cefepime for Injection with the volume of water specified in the labeling [NOTEUsing a suitable hypodermic needle and syringe, withdraw the entire contents of the vial, and dilute in Mobile phase.]
Impurity Name Cefepime Cefepime related compound A Cefepime related compound B Any other impurity
USP 32
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 1.7 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C19H24N6 O5S2 in the container of Cefepime for Injection: Result = (rU/rS) (CS/CU) P 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cefepime Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of cefepime CU hydrochloride in the Sample solution (mg/mL) P = content of cefepime in the USP Cefepime Hydrochloride RS (g/mg) Acceptance criteria: 90.0%115.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements rU
Official Monographs / Cefepime 131

= peak response of N-methylpyrrolidine from the Sample solution = peak response of N-methylpyrrolidine from the rS Standard solution = concentration of N-methylpyrrolidine in the CS Standard solution (mg/mL) = nominal concentration of cefepime in the CU Sample solution (mg/mL) Acceptance criteria: NMT 1.0% [NOTECefepime from the Sample solution elutes as a broad peak at 55 min. To minimize equilibration time at the start of the next day, it is recommended that the detector be turned on the night before and that Mobile phase be pumped through the system overnight at a flow rate of 0.2 mL/min. After every Sample solution injection, it is recommended that the chromatograph be flushed with Column rinse solution for 30 min at a flow rate of 1 mL/min to remove cefepime from the column and that the system then be switched back to Mobile phase at a flow rate of 1 mL/min for reequilibration.] PROCEDURE 2: RELATED COMPOUNDS Solution A: 0.68 mg/mL of monobasic potassium phosphate Solution B: Acetonitrile and Solution A (1:9) Adjust with potassium hydroxide or phosphoric acid to a pH of 5.0. Solution C: Acetonitrile and Solution A (1:1) Adjust with potassium hydroxide or phosphoric acid to a pH of 5.0. Mobile phase: See the gradient table below.
IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF N-METHYLPYRROLIDINE Mobile phase: Acetonitrile and 0.01 N nitric acid (1:100) Column rinse solution: Transfer 5.0 mL of nitric acid to a 1-L volumetric flask. Dilute with water to volume. Transfer this solution to an appropriate flask, and add 1 L of acetonitrile. Standard stock solution: 1.6 mL of Nmethylpyrrolidine/100 mL Standard solution: Standard stock solution and 0.01 N nitric acid (1:24) [NOTEThis solution contains 0.05 mg/mL of Nmethylpyrrolidine.] Sample solution: 10 mg/mL of cefepime from one container of Cefepime for Injection with the volume of water specified in the labeling, in 0.05 N nitric acid [NOTEInject this solution immediately.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Conductivity [NOTEThe typical background conductance is 3500 S.] Column: 4.6-mm 5-cm; 5-m packing L52 Guard column: 4.4-mm 5-cm; packing L17, placed between the pump and the injector Flow rate: 1 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Retention time: NLT 8 min for N-methylpyrrolidine Relative standard deviation: NMT 5.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of N-methylpyrrolidine in the portion of Cefepime for Injection taken: Result = (rU/rS) (CS/CU) 100
System suitability solution: 1.4 mg/mL of USP Cefepime Hydrochloride System Suitability RS in Solution B Sample solution: Constitute one container of Cefepime for Injection with a volume of Solution B equivalent to the volume of solvent specified in the labeling, and shake to dissolve. Transfer the constituted solution to a volumetric flask, and dilute with Solution B to obtain a solution having a concentration of 2 mg of cefepime/mL. [NOTEInject this solution immediately, or store in a refrigerator and inject within 12 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution and Sample solution Suitability requirements Resolution: NLT 5 between cefepime and cefepime related compound A; NLT 10 between cefepime related compound A and cefepime related compound B, System suitability solution Capacity factor, k: NLT 0.6, Sample solution Column efficiency: NLT 4000 theoretical plates, Sample solution
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USP 32
DEFINITION Cefixime contains the equivalent of NLT 950 g and NMT 1030 g of C16H15N5O7S2/mg, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197K Sample: Dissolve 5 mg of it by trituration in 2 mL of methanol and evaporate with the aid of gentle heat to dryness. ASSAY PROCEDURE Solution A: 25 mL of 0.4 M tetrabutylammonium hydroxide solution diluted to 1000 mL with water, adjusted with 1.5 M phosphoric acid to a pH of 6.5 Solution B: Dissolve 6.8 g of monobasic potassium phosphate in water to 500 mL. Solution C: Dissolve 7.1 g of anhydrous dibasic sodium phosphate in water to 500 mL. Buffer: Adjust a volume of Solution C with a sufficient volume of Solution B to a pH of 7.0. Mobile phase: Acetonitrile and Solution A (1:3) System suitability solution: 1 mg/mL of USP Cefixime RS in water Heat this solution at 95 in an oil bath for 45 min, cool, and use promptly. Standard solution: 0.2 mg/mL of USP Cefixime RS in Buffer Use this solution promptly. Sample solution: 0.22 mg/mL Cefixime in Buffer Use this solution promptly. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 12.5-cm; 4-m packing L1 Temperature: 40 Flow rate: Adjusted so that the retention time of cefixime is 10 min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTERelative retention times are about 0.9 for cefixime (E)-isomer and 1.0 for cefixime.] Suitability requirements Resolution: NLT 2.0 between cefixime and cefixime (E)isomer, System suitability solution Column efficiency: NLT 4000 theoretical plates, Standard solution, when calculated: 5.545(t/Wh/2)2 Tailing factor: For the analyte peak, between 0.9 and 2.0, Standard solution, when calculated: W0.1/2f = width of peak of 10% height W0.1 Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C16H15N5O7S2 in each mg of Cefixime taken: Result = (rU/rS) (CS/CU) F rU rS CS = peak response of cefixime from the Sample solution = peak response of cefixime from the Standard solution = concentration of cefixime in the Standard solution (mg/mL)
Tailing factor: NMT 1.5, Sample solution Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Cefepime for Injection taken: Result = (rU/rT) 100 rU = peak response for each impurity = sum of the responses for all the peaks rT Acceptance criteria: See Impurity Table 1.
Impurity Table 1 Relative Retention Time 2.7 4.3 1.0 Acceptance Criteria, NMT (%) 0.5 0.5 0.5
Name Cefepime related compound A Cefepime related compound B Any other impurity Cefepime
SPECIFIC TESTS INJECTIONS, Constituted Solutions 1: At the time of use, it meets the requirements. BACTERIAL ENDOTOXINS TEST 85: NMT 0.06 USP Endotoxin Unit/mg of cefepime STERILITY TESTS 71: Meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration PH 791: 4.06.0, in a solution containing 100 mg/mL of cefepime WATER DETERMINATION, Method I 921: NMT 4.0% OTHER REQUIREMENTS: Meets the requirements for Injections 1, Labels and Labeling ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant as described under Injections 1, Containers for Sterile Solids, and store in a refrigerator or at controlled room temperature. Store reconstituted powder in a refrigerator for NMT 7 days. LABELING: Label it to indicate that it is to be diluted with a suitable parenteral vehicle before intravenous infusion. USP REFERENCE STANDARDS 11 USP Cefepime Hydrochloride RS USP Cefepime Hydrochloride System Suitability RS USP Endotoxin RS
Cefixime
(Comment on this Monograph)id=m13990=Cefixime=Ca-ChlMonos.pdf)
507.50 C16H15N5O7S2 3H2O 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2amino-4-thiazolyl)[(carboxymethoxy)imino]acetyl]amino]-3ethenyl-8-oxo-, trihydrate, [6R-[6,7(Z)]]-; (6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-8-oxo-3-vinyl-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 72-(Z)-[O(carboxymethyl)oxime]trihydrate [79350-37-1]. Anhydrous 453.46.
USP 32
= concentration of Cefixime in the Sample solution (mg/mL) F = conversion factor, 1000 g/mg Acceptance criteria: 9501030 g/mg IMPURITIES Organic Impurities PROCEDURE 1 Solution A, Solution B, Solution C, Buffer, Mobile phase, System suitability solution, Standard solution and Sample solution: Proceed as directed in the Assay. Chromatographic system: Proceed as directed in the Assay. System suitability: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Cefixime taken: Result = (rU/rS) P F 100 = peak area for each impurity = cefixime peak area = potency of cefixime calculated in the Assay (g/mg) F = conversion factor, 0.001 mg/g Acceptance criteria Individual impurities: NMT 1.0% of any individual impurity is found. Total impurities: NMT 2.0% SPECIFIC TESTS SPECIFIC ROTATION 781S: 75 to 88 Sample solution: 10 mg/mL, in sodium bicarbonate solution (2 in 100) CRYSTALLINITY 695: Meets the requirements PH 791: 2.64.1 Sample solution: Contains the equivalent of 0.7 mg/mL of cefixime WATER DETERMINATION, Method I 921: 9.0%12.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label to indicate that it is the trihydrate form. Where the quantity of Cefixime is indicated in the labeling of any preparation containing Cefixime, this shall be understood to be in terms of anhydrous cefixime (C16H15N5O7S2). USP REFERENCE STANDARDS 11 USP Cefixime RS rU rS P CU
Official Monographs / Cefixime 133

ASSAY PROCEDURE Solution A: 0.4 M tetrabutylammonium hydroxide solution and water (1:39) Adjust with 1.5 M phosphoric acid to a pH of 6.5. Solution B: 13.6 mg/mL of monobasic potassium phosphate Solution C: 14.2 mg/mL of anhydrous dibasic sodium phosphate Adjust a volume of this solution with a sufficient volume of Solution B to a pH of 7.0. Mobile phase: Acetonitrile and Solution A (1:3) System suitability solution: 1 mg/mL of USP Cefixime RS [NOTEHeat this solution at 95 in an oil bath for 45 min, cool, and use promptly.] Standard solution: 0.2 mg/mL of USP Cefixime RS in Solution C [NOTEUse this solution promptly.] Sample solution: Constitute Cefixime for Oral Suspension as directed in the labeling. Quantitatively dilute a measured volume of the suspension thus obtained, freshly mixed and free from air bubbles, with Solution C to obtain a solution having a nominal concentration of 0.2 mg of cefixime/mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 12.5-cm; 4-m packing L1 Temperature: 40 Flow rate: 10 min Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for cefixime (E)-isomer and cefixime are about 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between cefixime and cefixime (E)isomer, System suitability solution Column efficiency: NLT 4000 theoretical plates when calculated: 5.545(t/Wh/2)2, Standard solution Tailing factor: NLT 0.9 and NMT 2.0 for the analyte peak when calculated: W0.1/2f = width of peak of 10% height, Standard solution W0.1 Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H15N5O7S2 in the constituted suspension prepared from the Cefixime for Oral Suspension: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response of cefixime from the Sample solution = peak response of cefixime from the Standard solution = concentration of USP Cefixime RS in the Standard solution (mg/mL) = nominal concentration of cefixime in the Sample solution (mg/mL)
Cefixime for Oral Suspension

(Comment on this Monograph)id=m13992=Cefixime for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cefixime for Oral Suspension is a dry mixture of Cefixime and one or more suitable diluents, flavors, preservatives, and suspending agents. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cefixime (C16H15N5O7S2)/mL when constituted as directed in the labeling. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
134
Cefixime / Official Monographs

90.0%120.0%
USP 32
Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for cefixime (E)-isomer and cefixime are about 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between cefixime and cefixime (E)isomer, System suitability solution Column efficiency: NLT 4000 theoretical plates for the Standard solution when calculated: 5.545(t/Wh/2)2 Tailing factor: NLT 0.9 and NMT 2.0 for the analyte peak when calculated: W0.1/2f W0.1 = width of peak of 10% height, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H15N5O7S2 in the portion of Cefixime Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of cefixime from the Sample solution rS = peak response of cefixime from the Standard solution = concentration of USP Cefixime RS in the Standard CS solution (mg/mL) CU = nominal concentration of cefixime in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION 711 Medium: 6.8 mg/mL of monobasic potassium phosphate in 1000 mL of water. Adjust with 1 N sodium hydroxide to a pH of 7.2; 900 mL. Apparatus 1: 100 rpm Time: 45 min Detector: UV 288 nm Standard solution: USP Cefixime RS in Medium Sample solutions: Sample per Dissolution 711. Dilute with Medium to concentration similar to that of the Standard solution [NOTEAn amount of methanol not to exceed 0.1% of the total volume of the Standard solution may be used to bring the USP Cefixime RS into solution prior to dilution with Medium, and the solution may be sonicated to ensure complete dissolution of the USP Cefixime RS.] Tolerances: NLT 75% (Q) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 10.0%
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for solids packaged in single-unit containers DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 2.54.5, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that the cefixime contained therein is in the trihydrate form. USP REFERENCE STANDARDS 11 USP Cefixime RS
Cefixime Tablets
(Comment on this Monograph)id=m13994=Cefixime Tablets=Ca-Chl-Monos.pdf) DEFINITION Cefixime Tablets contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of cefixime (C16H15N5O7S2). IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.4 M tetrabutylammonium hydroxide solution and water (1:39) Adjust with 1.5 M phosphoric acid to a pH of 6.5. Solution B: 13.6 mg/mL of monobasic potassium phosphate Solution C: 14.2 mg/mL of anhydrous dibasic sodium phosphate Adjust a volume of this solution with a sufficient volume of Solution B to a pH of 7.0. Mobile phase: Acetonitrile and Solution A (1:3) System suitability solution: 1 mg/mL of USP Cefixime RS [NOTEHeat this solution at 95 in an oil bath for 45 min, cool, and use promptly.] Standard solution: 0.2 mg/mL of USP Cefixime RS in Solution C [NOTEUse this solution promptly.] Sample stock solution: Transfer powdered tablets equivalent to about 400 mg of cefixime (NLT 20 tablets), to a 100-mL volumetric flask, add 75 mL of Solution C and sonicate. Dilute with Solution C to volume, mix, and centrifuge. Sample solution: Nominal concentration of 0.2 mg/mL of Sample stock solution in Solution C Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 12.5-cm; 4-m packing L1 Temperature: 40 Flow rate: Adjust it so that the retention time of cefixime is about 10 min.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Tablets to indicate that the cefixime contained therein is in the trihydrate form. USP REFERENCE STANDARDS 11 USP Cefixime RS
USP 32
Official Monographs / Cefmenoxime 135

Mode: LC Detector: UV 254 nm Column: 4-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.3 between the phthalimide and cefmenoxime peaks Column efficiency: NLT 1200 theoretical plates from the cefmenoxime peak Tailing factor: NMT 1.6 for the cefmenoxime peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity of C16H17N9O5S3 g in each mg of Cefmenoxime Hydrochloride taken: Result = (RU/RS) (CS/CU) P = peak area ratio of cefmenoxime to the internal standard from the Sample solution = peak area ratio of cefmenoxime to the internal RS standard from the Standard solution = concentration of USP Cefmenoxime CS Hydrochloride RS in the Standard solution (mg/mL) = concentration of Cefmenoxime Hydrochloride in CU the Sample solution (mg/mL) P = designated C16H17N9O5S3 content of USP Cefmenoxime Hydrochloride RS (g/mg) Acceptance criteria: 8691015 g/mg RU SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PYROGEN TEST 151: Where the label states that it is sterile or must be subjected to further processing during the solution of injectable dosage forms, it meets the requirements, the test dose being 1.0 mL/kg of a solution in pyrogen-free sodium carbonate solution (prepared by dissolving 14.0 g of sodium carbonate, previously heated at 170 for NLT 4 h, in 1000 mL of sterile water for injection) containing 60 mg/mL. STERILITY TESTS 71: Where the label states that it is sterile, it meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration, except to add 2.0 g of sodium carbonate (previously sterilized by heating at 180 for 2 h) to each 100 mL of Fluid A. WATER DETERMINATION, Method I 921: NMT 1.5% Sample solution: Prepared as directed for a hygroscopic specimen, except use 20 mL of a mixture of formamide (previously dried over anhydrous sodium sulfate for 24 h) and methanol (2:1), instead of methanol, to dissolve the specimen, use two 5-mL portions of the same formamide and methanol mixture to rinse the container, and determine the water content of the formamide and methanol mixture. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or
Cefmenoxime Hydrochloride
(Comment on this Monograph)id=m14000=Cefmenoxime Hydrochloride=Ca-Chl-Monos.pdf)
1059.58 (C16H17N9O5S3)2 HCl 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2amino-4-thiazolyl)(methoxyimino)acetyl]amino]-3-[[(1methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, hydrochloride (2:1), [6R-[6,7(Z)]]-; (6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-3-[[(1methyl-1H-tetrazol-5-yl)-thio]methyl]-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid 72-(Z)-(Omethyloxime), hydrochloride (2:1) [75738-58-8]. DEFINITION Cefmenoxime Hydrochloride contains the equivalent of NLT 869 g and NMT 1015 g of C16H17N9O5S3/mg, calculated on the anhydrous basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Solution: 25 g/mL in Solution A prepared as directed in the Assay. B. The retention time of the cefmenoxime peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Solution A: 6.4 mg/mL of monobasic potassium phosphate and 18.9 mg/mL of dibasic sodium phosphate in water Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1. Mobile phase: Acetonitrile, glacial acetic acid, and water (10:1:50) Filter through a suitable filter of 0.5 m or finer porosity. Internal standard solution: 1.5 mg/mL of phthalimide in methanol Standard solution: Transfer 50 mg of USP Cefmenoxime Hydrochloride RS to a 50-mL volumetric flask, add 10 mL of pH 6.8 buffer, and dissolve by swirling. Dilute with Mobile phase to volume. Transfer 4.0 mL of this solution to a second 50-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with Mobile phase to volume. [NOTEThis solution contains the equivalent of 80 g/mL of C16H17N9O5S3.] Sample stock solution: Transfer 50 mg of Cefmenoxime Hydrochloride RS to a 50-mL volumetric flask, add 10 mL of pH 6.8 buffer, and dissolve by swirling. Dilute with Mobile phase to volume. Transfer 4.0 mL of this solution to a second 50-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with Mobile phase to volume. Sample solution: 2.0 mL of Sample stock solution and 10.0 mL of Internal standard solution Dilute with Mobile phase to 25.0 mL. Chromatographic system (See Chromatography 621, System Suitability.)
136
Cefmenoxime / Official Monographs
USP 32
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.3 between the phthalimide and the cefmenoxime peaks Column efficiency: NLT 1200 theoretical plates from the cefmenoxime peak when calculated based on the width of the peak at half height Tailing factor: NMT 1.6 for the cefmenoxime peak Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution A or Sample solution B, and Standard solution [NOTEUse peak areas where peak responses are indicated.] Calculate the percentage of C16H17N9O5S3 withdrawn from the container or in the portion of constituted solution taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the cefmenoxime peak to the internal standard peak from the Sample solution = peak response ratio of the cefmenoxime peak to RS the internal standard peak from the Standard solution = concentration of USP Cefmenoxime CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of cetmenoxime in CU Sample solution A or Sample solution B (mg/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. PYROGEN TEST 151: It meets the requirements, the test dose being 1.0 mL/kg of a solution of Cefmenoxime for Injection in Sterile Water for Injection having a concentration of 60 mg of cefmenoxime/mL. STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration PH 791: 6.47.9, in a solution containing the equivalent of 100 mg/mL of cefmenoxime LOSS ON DRYING 731: Dry 100 mg in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 1.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids USP REFERENCE STANDARDS 11 USP Cefmenoxime Hydrochloride RS RU
must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefmenoxime Hydrochloride RS
Cefmenoxime for Injection

(Comment on this Monograph)id=m14002=Cefmenoxime for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefmenoxime for Injection contains an amount of Cefmenoxime Hydrochloride equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of cefmenoxime (C16H17N9O5S3). It may contain sodium carbonate. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Solution: 25 g/mL in Solution A (prepared as directed in the Assay) B. The retention time of the major cefmenoxime peak in the chromatogram of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Solution A: 6.4 mg/mL of monobasic potassium phosphate and 18.9 mg/mL of dibasic sodium phosphate in water Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1. Mobile phase: Acetonitrile, glacial acetic acid, and water (10:1:50) Filter through a suitable filter of 0.5 m or finer porosity. Internal standard solution: 1.5 mg/mL of phthalimide in methanol Standard stock solution: 1 mg/mL of USP Cefmenoxime Hydrochloride RS in Solution A and Mobile phase (1:4) Standard solution: 2.0 mL of Standard stock solution and 10.0 mL of Internal standard solution Dilute with Mobile phase to 25.0 mL. [NOTEThis solution contains the equivalent of 80 g/mL of cefmenoxime.] Sample stock solution A (where it is represented as being in a single-dose container): 1 mg/mL of cefmenoxime from a container of Cefmenoxime for Injection in a volume of water corresponding to the volume of diluent specified in the labeling [NOTEWithdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe.], and quantitatively diluted with water. Sample solution A: 2.0 mL of Sample stock solution A and 10.0 mL of Internal standard solution Dilute with Mobile phase to 25.0 mL. [ NOTEThis solution contains the equivalent of 80 g/mL of cefmenoxime.] Sample stock solution B (where the label states the quantity of cefmenoxime in a given volume of constituted solution): 1 mg/mL of cefmenoxime from a container of Cefmenoxime for Injection in a volume of water equivalent to the volume of diluent specified in the labeling, and quantitatively diluted with water Sample solution B: 2.0 mL of Sample stock solution B and 10.0 mL of Internal standard solution Dilute with Mobile phase to 25.0 mL. [NOTEThis solution contains the equivalent of 80 g/mL of cefmenoxime.]
USP 32
Official Monographs / Cefmetazole 137

Analysis Samples: Sample solution and Standard solution Calculate the quantity, in g, of C15H17N7O5S3 in each mg of Cefmetazole taken: Result = (rU/rS) (CS/CU) = peak response of cefmetazole from the Sample solution rS = peak response of cefmetazole from the Standard solution CS = concentration of USP Cefmetazole RS in the Standard solution (g/mL) CU = nominal concentration of cefmetazole in the Sample solution (g/mL) Acceptance criteria: 9701030 g/mg SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5% rU
Cefmetazole
(Comment on this Monograph)id=m14011=Cefmetazole=CaChl-Monos.pdf)
C15H17N7O5S3 471.52 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[ [(cyanomethyl)thio]acetyl]amino]-7-methoxy-3-[[(1methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, (6R-cis)-; (6R,7S)-7-[2-[(Cyanomethyl)thio]acetamido]-7-methoxy-3-[[(1methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylic acid. [56796-20-4]. DEFINITION Cefmetazole contains NLT 970 g and NMT 1030 g of cefmetazole (C15H17N7O5S3)/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: To 5.75 g of monobasic ammonium phosphate in 700 mL of water, add 3.2 mL of a 40% solution of tetrabutylammonium hydroxide, 280 mL of methanol, and 25 mL of tetrahydrofuran. Adjust with phosphoric acid to a pH of 4.5 0.1, and pass through a filter having a 0.5-m or finer porosity. System suitability stock solution: 1 mg/mL of USP Cefmetazole RS in 0.01 N sodium hydroxide [NOTEHeat at 95 for 10 min.] System suitability solution: Mix 1 mL of the System suitabiity stock solution and 2 mL of the Standard solution. Dilute with Mobile phase to 20 mL. [NOTEThis solution contains cefmetazole and cefmetazole lactone (resolution compound).] Standard solution: 200 g/mL of cefmetazole by quantitatively dissolving USP Cefmetazole RS in Mobile phase [NOTEUse this solution within 10 min.] Sample solution: 200 g/mL of Cefmetazole in Mobile phase [NOTEUse this solution within 10 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 3.0 between cefmetazole and cefmetazole lactone, System suitability solution Column efficiency: NLT 1250 theoretical plates, Standard solution Tailing factor: 0.941.6, Standard solution Relative standard deviation: NMT 2.0%, Standard solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefmetazole RS
Cefmetazole Injection
(Comment on this Monograph)id=m14013=Cefmetazole Injection=Ca-Chl-Monos.pdf) DEFINITION Cefmetazole Injection is a sterile isoosmotic solution of Cefmetazole and Sodium Citrate in Water for Injection. It contains one or more buffer substances and a tonicityadjusting agent. It contains NLT 90.0% and NMT 120.0% of the labeled amount of cefmetazole (C15H17N7O5S3). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: To 5.75 g of monobasic ammonium phosphate in 700 mL of water, add 3.2 mL of a 40% solution of tetrabutylammonium hydroxide, 280 mL of methanol, and 25 mL of tetrahydrofuran, and mix. Adjust with phosphoric acid to a pH of 4.5 0.1, and pass through a filter having a 0.5-m or finer porosity. System suitability stock solution: 1 mg/mL of USP Cefmetazole RS in 0.01 N sodium hydroxide [NOTEHeat at 95 for 10 min.] System suitability solution: System suitability stock solution, Standard solution, and Mobile phase (1:2:17) [NOTEThis solution contains cefmetazole and cefmetazole lactone (resolution compound).] Standard solution: 200 g/mL of cefmetazole by quantitatively dissolving USP Cefmetazole RS in Mobile phase [NOTEUse this solution within 10 min.] Sample solution: Equivalent to 200 g/mL of cefmetazole from Ceftemetazole Injection Allow the contents of a container of Injection to thaw, and mix the resultant solution in Mobile phase. [NOTEUse this solution within 10 min.] Chromatographic system (See Chromatography 621, System Suitability.)
138
Cefmetazole / Official Monographs

USP Endotoxin RS
USP 32
Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 3.0 between cefmetazole and cefmetazole lactone, System suitability solution Column efficiency: NLT 1250 theoretical plates, Standard solution Tailing factor: 0.941.6, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the percentage of C15H17N7O5S3 in each mL of Cefmetazole Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of cefmetazole from the Sample solution rS = peak response of cefmetazole from the Standard solution = concentration of USP Cefmetazole RS in the CS Standard solution (g/mL) = nominal concentration of cefmetazole taken to CU prepare the Sample solution (g/mL) Acceptance criteria: 90.0%120.0% rU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin Unit/mg of cefmetazole STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration, except to use water instead of Fluid A. PH 791: 4.26.2 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements under Injections 1, Labeling. The label states that it is to be thawed just before use, describes the conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cefmetazole RS
Cefmetazole Sodium
(Comment on this Monograph)id=m14014=Cefmetazole Sodium=Ca-Chl-Monos.pdf)
493.53 C15H16N7NaO5S3 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[ [(cyanomethyl)thio]acetyl]amino]-7-methoxy-3-[[(1methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, monosodium salt, (6R-cis)-; Sodium (6R,7S)-7-[2-[(cyanomethyl)thio]acetamido]-7methoxy-3-[[(1-methyl-1 H-tetrazol-5-yl)thio]methyl]-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate [56796-39-5]. DEFINITION Cefmetazole Sodium contains the equivalent of NLT 860 g and NMT 1003 g of cefmetazole (C15H17N7O5S3)/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: To 5.75 g of monobasic ammonium phosphate in 700 mL of water, add 3.2 mL of a 40% solution of tetrabutylammonium hydroxide, 280 mL of methanol, and 25 mL of tetrahydrofuran. Adjust with phosphoric acid to a pH of 4.5 0.1, and pass through a filter having a 0.5-m or finer porosity. System suitability stock solution: 1 mg/mL of USP Cefmetazole RS in 0.01 N sodium hydroxide [NOTEHeat at 95 for 10 min.] System suitability solution: Mix 1 mL of the System suitability stock solution and 2 mL of the Standard solution. Dilute with Mobile phase to volume. [NOTEThis solution contains cefmetazole and cefmetazole lactone (resolution compound).]
USP 32
Standard solution: 200 g/mL of cefmetazole by quantitatively dissolving USP Cefmetazole RS in Mobile phase [NOTEUse this solution within 10 min.] Sample solution: 210 g/mL of cefmetazole sodium in Mobile phase [NOTEUse this solution within 10 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 3.0 between cefmetazole and cefmetazole lactone, System suitability solution Column efficiency: NLT 1250 theoretical plates, Standard solution Tailing factor: 0.941.6, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample solution and Standard solution Calculate the quantity, in g, of C15H17N7O5S3/mg of Cefmetazole Sodium taken: Result = (rU/rS) (CS/CU) = peak response of cefmetazole from the Sample solution = peak response of cefmetazole from the Standard rS solution = concentration of USP Cefmetazole RS in the CS Standard solution (g/mL) = nominal concentration of cefmetazole sodium in CU the Sample solution (g/mL) Acceptance criteria: 8601003 g/mg SPECIFIC TESTS PH 791: 4.26.2, in a solution (1 in 10) WATER DETERMINATION, Method I 921: NMT 0.5% STERILITY TESTS 71: Where the label states that Cefmetazole Sodium is sterile, it meets the requirements.Test for Sterility of the Product to Be Examined, Membrane Filtration BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefmetazole Sodium is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefmetazole RS USP Endotoxin RS rU
Official Monographs / Cefmetazole 139

IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 5.75 g of monobasic ammonium phosphate in 700 mL of water. Add 3.2 mL of a 40% solution of tetrabutylammonium hydroxide, 280 mL of methanol, and 25 mL of tetrahydrofuran, and mix. Adjust with phosphoric acid to a pH of 4.5 0.1, and pass through a filter having a 0.5-m or finer porosity. System suitability stock solution: 1 mg/mL of USP Cefmetazole RS in 0.01 N sodium hydroxide [NOTEHeat at 95 for 10 min.] System suitability solution: 1.0 mL of System suitability stock solution and 2.0 mL of Standard solution Dilute with Mobile phase to 20.0 mL. [NOTEThis solution contains cefmetazole and cefmetazole lactone (resolution compound).] Standard solution: 200 g/mL of cefmetazole by quantitatively dissolving USP Cefmetazole RS in Mobile phase [NOTEUse this solution within 10 min.] Sample solution A: Where it is represented as being in a single-dose container, 200 g/mL of cefmetazole from a container of Cefmetazole for Injection in a volume of water, corresponding to the volume of diluent specified in the labeling, and quantitatively diluted with Mobile Phase [NOTEWithdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe.] [NOTEUse this solution within 10 min.] Sample solution B: Where the label states the quantity of cefmenoxime in a given volume of constituted solution, 200 g/mL of cefmetazole from a container of Cefmetazole for Injection in a volume of water, equivalent to the volume of diluent specified in the labeling, and quantitatively diluted with Mobile phase [NOTEUse this solution within 10 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 3.0 between cefmetazole and cefmetazole lactone, System suitability solution Column efficiency: NLT 1250 theoretical plates, Standard solution Tailing factor: 0.941.6, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample solution A or Sample solution B, and Standard solution Calculate the percentage of C15H17N7O5S3 withdrawn from the container, or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of cefmetazole from the Sample solution
Cefmetazole for Injection

(Comment on this Monograph)id=m14016=Cefmetazole for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefmetazole for Injection contains an amount of Cefmetazole Sodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cefmetazole (C15H17N7O5S3).
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Cefmetazole / Official Monographs

rS
USP 32
Heat on a steam bath for 30 min, and cool. This System suitability solution contains a mixture of cefonicid and desacetyl cefonicid. Standard solution: Equivalent to 200 g/mL of cefonicid from USP Cefonicid Sodium RS idissolved in Mobile phase Sample solution: 200 g/mL of Cefonicid Sodium in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.1 between the cefonicid and the desacetyl cefonicid peaks, System suitability solution Column efficiency: NLT 1500 from the analyte peak theoretical plates, Standard solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C18H18N6O8S3 per mg of Cefonicid Sodium taken: Result = (rU/rS) (CS/CU) Mr1/Mr2 F = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cefonicid Sodium RS in the Standard solution (g/mL) = nominal concentration of cefonicid sodium in CU the Sample solution (g/mL) = molecular weight of cefonicid Mr1 = molecular weight of cefonicid sodium Mr2 F = conversion factor, 1000 g/mg Acceptance criteria: 832970 g/mg SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: Between 37 and 47 Sample solution: 10 mg/mL, in methanol PH 791: 3.56.5, in a solution (1 in 20) WATER DETERMINATION, Method I 921: NMT 5.0% STERILITY TESTS 71: Where the label states that Cefonicid Sodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefonicid Sodium is sterile or it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.35 USP Endotoxin Unit/mg of cefonicid. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefonicid Sodium RS USP Endotoxin RS rU rS CS
= peak response of cefmetazole from the Standard solution = concentration of USP Cefmetazole RS in the CS Standard solution (g/mL) = nominal concentration of cefmetazole in Sample CU solution A or Sample solution B (g/mL) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: requirements. It meets the
SPECIFIC TESTS PH 791: 4.26.2, in a solution (1 in 10) WATER DETERMINATION, Method I 921: NMT 0.5% BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin Unit/mg of cefmetazole STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. OTHER REQUIREMENTS: It meets the requirements under Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefmetazole RS USP Endotoxin RS
Cefonicid Sodium
(Comment on this Monograph)id=m14031=Cefonicid Sodium=Ca-Chl-Monos.pdf)
586.53 C18H16N6Na2O8S3 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7[(hydroxyphenylacetyl)amino]-8-oxo-3-[[[1-(sulfomethyl)-1Htetrazol-5-yl]thio]methyl]disodium salt, [6R-[6,7 (R*)]]-; (6R,7R)-[7-[(R)-Mandelamido]-8-oxo-3-[[[1-(sulfomethyl)-1Htetrazol-5-yl]thio]methyl]-5-thia-1-azabicyclo[4.2.0]oct-2ene-2-carboxylic acid, disodium salt [61270-78-8]. DEFINITION Cefonicid Sodium contains the equivalent of NLT 832 g and NMT 970 g of cefonicid (C18H18N6O8S3)/mg, calculated on the anhydrous basis. IDENTIFICATION A. The retention time of the major peak for cefonicid of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets the requirements. ASSAY PROCEDURE Mobile phase: Methanol, 0.2 M monobasic ammonium phosphate, and water (5:2:33) Pass through a filter having a 0.5-m or finer porosity. System suitability solution: 200 g/mL of USP Cefonicid Sodium RS in Mobile phase
USP 32
Official Monographs / Cefoperazone 141

= peak response from the Standard solution = concentration of USP Cefonicid Sodium RS in the Standard solution (g/mL) CU = nominal concentration of cefonicid in Sample solution A or Sample solution B (g/mL) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: requirements. It meets the rS CS
Cefonicid for Injection

(Comment on this Monograph)id=m14035=Cefonicid for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefonicid for Injection contains an amount of Cefonicid Sodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cefonicid (C18H18N6O8S3). IDENTIFICATION A. The retention time of the major peak for cefonicid in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191 It meets the requirements. ASSAY PROCEDURE Mobile phase: Methanol, 0.2 M monobasic ammonium phosphate, and water (5:3:33) Pass through a filter having a 0.5-m or finer porosity. System suitability solution: 200 g/mL of USP Cefonicid Sodium RS in Mobile phase [NOTEHeat on a steam bath for 30 min, and cool. This System suitability solution contains a mixture of cefonicid and desacetyl cefonicid.] Standard solution: 200 g/mL of cefonicid by quantitatively dissolving USP Cefonicid Sodium RS in Mobile phase Sample solution A (where it is represented as being in a single-dose container): 200 g/mL of cefonicid in Mobile phase prepared as follows. Using a suitable hypodermic needle and syringe, withdraw all of the withdrawable contents from a container of Cefonicid for Injection in a volume of water corresponding to the volume of solvent specified in the labeling, and dilute quantitatively with Mobile phase. Sample solution B (where the label states the quantity of cefonicid in a given volume of constituted solution): 200 g/mL of cefonicid in Mobile phase, from a container of Cefonicid for Injection in a volume of water corresponding to the volume of solvent specified in the labeling, quantitatively diluted with Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.1 between the cefonicid and the desacetyl cefonicid peaks, System suitability solution Column efficiency: NLT 1500 theoretical plates from the analyte peak, Standard solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample solution A or Sample solution B, and Standard solution Calculate the percentage of C18H18N6O8S3 withdrawn from the container, or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100 rU = peak response from the Sample solution
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 37 to 47 Sample solution: 10 mg/mL, in methanol PH 791: 3.56.5, in a solution (1 in 20) WATER DETERMINATION, Method I 921: NMT 5.0% CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: NMT 0.35 USP Endotoxin Unit/mg of cefonicid STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. OTHER REQUIREMENTS: It meets the requirements for Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefonicid Sodium RS USP Endotoxin RS
Cefoperazone Sodium
(Comment on this Monograph)id=m14039=Cefoperazone Sodium=Ca-Chl-Monos.pdf)
667.65 C25H26N9NaO8S2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[[(4ethyl-2,3-dioxo-1-piperazinyl)carbonyl]amino](4hydroxyphenyl)acetyl]amino]-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-, monosodium salt, [6R-[6,7 (R*)]]-; Sodium (6R,7R)-7-[(R)-2-(4-ethyl-2,3-dioxo-1piperazinecarboxamido)-2-(p-hydroxyphenyl)acetamido-3-[ [(1-methyl-H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylate [62893-20-3]. DEFINITION Cefoperazone Sodium contains the equivalent of NLT 870 g and NMT 1015 g of cefoperazone (C25H27N9O8S2)/mg, calculated on the anhydrous basis. IDENTIFICATION A. The retention time of the major peak for cefoperazone of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements
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Cefoperazone / Official Monographs
USP 32
ASSAY PROCEDURE Solution A: Place 14 mL of triethylamine and 5.7 mL of glacial acetic acid in a 100-mL volumetric flask, and dilute with water to volume. Mobile phase: Acetonitrile, 1 N acetic acid, Solution A, and water (120:2.8:1.2:876) Filter through a membrane filter of 1-m or finer porosity. Standard solution: 160 g/mL of cefoperazone from USP Cefoperazone Dihydrate RS in Mobile phase Sample solution: 160 g/mL of Cefoperazone Dihydrate in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Sample: Sample solution and Standard solution Calculate the quantity, in g of cefoperazone/mg, of Cefoperazone Sodium: Result = (rU/rS) (CS/CU) F = peak response from the Sample solution = peak response from the Standard solution = concentration of cefoperazone in the Standard solution (g/mL) = concentration of Cefoperazone Sodium in the CU Sample solution (g/mL) F = correction factor, 1000 g/mg Acceptance criteria: 8701015 g/mg rU rS CS SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 4.56.5, in a solution (1 in 4) WATER DETERMINATION, Method I 921: NMT 5.0% STERILITY TESTS 71: Where the label states that Cefoperazone Sodium is sterile, it meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefoperazone Sodium is sterile or it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.20 USP Endotoxin Unit/mg of cefoperazone. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefoperazone Dihydrate RS USP Endotoxin RS
Cefoperazone Injection
(Comment on this Monograph)id=m14041=Cefoperazone Injection=Ca-Chl-Monos.pdf) DEFINITION Cefoperazone Injection is a sterile solution of Cefoperazone Sodium and a suitable osmolality adjusting substance in Water for Injection. It may contain a suitable buffer. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cefoperazone (C25H27N9O8S2). IDENTIFICATION The retention time of the major peak for cefoperazone from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Triethylamine, glacial acetic acid, and water (14:5.7:80.3) Mobile phase: Acetonitrile, 1 N acetic acid, Solution A, and water (120:2.8:1.2:876) Filter through a membrane filter of 1-m or finer porosity. Standard solution: 160 g/mL of cefoperazone by quantitatively dissolving USP Cefoperazone Dihydrate RS in Mobile phase Sample solution: 160 g/mL of cefoperazone from the volume of Injection in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of C25H27N9O8S2 in the volume of Injection: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cefoperazone Dihydrate RS in the Standard solution (g/mL) = nominal concentration of cefoperazone in the CU Sample solution (g/mL) Acceptance criteria: 90.0%120.0% rU rS CS SPECIFIC TESTS PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 4.56.5 BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Unit/mg of cefoperazone
USP 32
STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements under Injections 1, Labeling. The label states that it is to be thawed just prior to use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cefoperazone Dihydrate RS USP Endotoxin RS
Official Monographs / Cefoperazone 143

Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution A or Sample solution B, and Standard solution Calculate the percentage of C25H27N9O8S2 withdrawn from the container, or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cefoperazone Dihydrate RS in the Standard solution (g/mL) = nominal concentration of cefoperazone in CU Sample solution A or Sample solution B (g/mL) Acceptance criteria: 90.0%120.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: requirements. It meets the
Cefoperazone for Injection

(Comment on this Monograph)id=m14047=Cefoperazone for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefoperazone for Injection contains an amount of Cefoperazone Sodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cefoperazone (C25H27N9O8S2). IDENTIFICATION A. The retention time of the major peak for cefoperazone from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets the requirements. ASSAY PROCEDURE Solution A: Triethylamine, glacial acetic acid, and water (14:5.7:80.3) Mobile phase: Acetonitrile, 1 N acetic acid, Solution A, and water (120:2.8:1.2:876) Pass through a membrane filter having a 1-m or finer porosity. Standard solution: 160 g/mL of cefoperazone by quantitatively dissolving USP Cefoperazone Dihydrate RS in Mobile phase Sample solution A (where it is represented as being in a single-dose container): 160 g/mL of cefoperazone in Mobile phase prepared as follows. Using a suitable hypodermic needle and syringe, withdraw all of the withdrawable contents from a container of Cefoperazone for Injection in a volume of water corresponding to the volume of solvent specified in the labeling, and dilute quantitatively with Mobile phase. Sample solution B (where the label states the quantity of cefoperazone in a given volume of constituted solution): 160 g/mL of cefoperazone in Mobile phase, from a container of Cefoperazone for Injection in a volume of water corresponding to the volume of solvent specified in the labeling, quantitatively diluted with Mobile phase.
SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Unit/mg of cefoperazone STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. PH 791: 4.56.5, in a solution (1 in 4) WATER DETERMINATION, Method I 921: NMT 5.0%, except that where it is in the freeze-dried form, the limit is NMT 2.0% OTHER REQUIREMENTS: It meets the requirements for Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefoperazone Dihydrate RS USP Endotoxin RS
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Ceforanide / Official Monographs

rU rS CS
USP 32
= peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ceforanide RS in the Standard solution (mg/mL) CU = concentration of Ceforanide taken to prepare Sample solution A, Sample solution B, or Sample solution C (mg/mL) Acceptance criteria: 9001050 g/mg SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: Where the label states that Ceforanide is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.25 USP Endotoxin Unit/mg of Ceforanide. STERILITY TESTS 71: Where the label states that Ceforanide is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration, except to dissolve 6 g of Ceforanide in Fluid A to each 1000 mL of which has been added 10 g of sterile Llysine, and to rinse the membrane with three 100-mL portions of Fluid D and one 100-mL portion of Fluid A. PH 791: 2.54.5, in a suspension containing 50 mg/mL WATER DETERMINATION, Method I 921: NMT 5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Ceforanide RS USP Endotoxin RS
Ceforanide
(Comment on this Monograph)id=m14072=Ceforanide=Ca-ChlMonos.pdf)
C20H21N7O6S2 519.56 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[2(aminomethyl)phenyl]acetyl]amino]-3-[[[1(carboxymethyl)-1H-tetrazol-5-yl]thio]methyl]-8-oxo-, (6Rtrans)-; (6R,7R)-7-[2-(-Amino-o-tolyl)acetamido]-3-[[[1(carboxymethyl)-1H-tetrazol-5-yl]thio]methyl]-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid; 7-[o-(Aminomethyl)phenylacetamido]-3-[[[1(carboxymethyl)-1H-tetrazol-5-yl]thio]methyl]-3-cephem-4carboxylic acid [60925-61-3]. DEFINITION Ceforanide contains NLT 900 g and NMT 1050 g of ceforanide (C20H21N7O6S2)/mg. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak for ceforanide of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 18 mL of tetrabutylammonium hydroxide solution (1 in 10) and 8.6 mL of 11 N potassium hydroxide Add the mixture to 700 mL of water. Mobile phase: To Solution A, add 200 mL of methanol, adjust with phosphoric acid to a pH of 7.0, and add water to obtain 1000 mL of solution. Filter, using a filter having a porosity of 1 m or finer. Standard solution: 1 mg/mL of USP Ceforanide RS in Mobile phase [NOTEUse this solution within 5 min.] Sample solution: 1 mg/mL of Ceforanide in Mobile phase [NOTEUse this solution within 5 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; 5- to 10-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: 1.85.0 Column efficiency: NLT 1900 theoretical plates from the analyte peak Tailing factor: NMT 1.2 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C20H21N7O6S2 in each mg of Ceforanide taken: Result = (rU/rS) (CS/CU)
Ceforanide for Injection

(Comment on this Monograph)id=m14073=Ceforanide for Injection=Ca-Chl-Monos.pdf) DEFINITION Ceforanide for Injection is a sterile mixture of Ceforanide and Llysine. It contains NLT 900 g and NMT 1050 g of ceforanide (C20H21N7O6S2)/mg on the L-lysinefree basis, and NLT 90.0% and NMT 115.0% of the labeled amount of C20H21N7O6S2. IDENTIFICATION A. The retention time of the major peak for L-lysine in the Sample solution corresponds to that of the Standard solution, as obtained in the test for Content of L-Lysine. B. The retention time of the major peak for ceforanide in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Mix 18 mL of tetrabutylammonium hydroxide solution (1 in 10) and 8.6 mL of 11 N potassium hydroxide, and add the mixture to 700 mL of water. Mobile phase: To Solution A, add 200 mL of methanol, adjust with phosphoric acid to a pH of 7.0, and add water to obtain 1000 mL of solution. Filter, using a filter having a porosity of 1 m or finer.
USP 32
Standard solution: 1 mg/mL of USP Ceforanide RS in Mobile phase [NOTEUse this solution within 5 min.] Sample solution A: 1 mg/mL of Ceforanide for Injection in Mobile phase [NOTEUse this solution within 5 min.] Sample solution B (where it is represented as being in a single-dose container): 1 mg/mL of ceforanide from a container of Ceforanide for Injection in a volume of water corresponding to the volume of solvent specified in the labeling Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute quantitatively and stepwise with Mobile phase. [NOTEUse this solution within 5 min.] Sample solution C (where the label states the quantity of ceforanide in a given volume of constituted solution): 1 mg/mL of ceforanide from a container of Ceforanide for Injection in a volume of water corresponding to the volume of solvent specified in the labeling Dilute measured volume of the constituted solution quantitatively and stepwise with Mobile phase. [NOTEUse this solution within 5 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; 5- to 10-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: 1.85.0 Column efficiency: NLT 1900 theoretical plates from the analyte peak Tailing factor: NMT 1.2 Relative standard deviation: NMT 1.5% Analysis 1 Samples: Standard solution and Sample solution A Calculate the g of C20H21N7O6S2 in each mg of Ceforanide for Injection taken: Result = (rU/rS) (CS/CU) P = peak response from Sample solution A = peak response from the Standard solution = concentration of USP Ceforanide RS in the Standard solution (mg/mL) = nominal concentration of ceforanide in Sample CU solution A (mg/mL) P = potency of USP Ceforanide RS (g/mg) Analysis 2 Samples: Standard solution and Sample solution B, or Sample solution C Calculate the percentage of label claim of C20H21N7O6S2 in the Ceforanide for Injection taken: Result = (rU/rS) (CS/CU) P F 100 = peak response from the appropriate Sample solution rS = peak response from the Standard solution = concentration of USP Ceforanide RS in the CS Standard solution (mg/mL) = nominal concentration of ceforanide in the CU appropriate Sample solution (mg/mL) P = potency of USP Ceforanide RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 9001050 g/mg of C20H21N7O6S2 (Llysinefree basis) and 90.0%115.0% of label claim of C20H21N7O6S2 rU rU rS CS
Official Monographs / Ceforanide 145

OTHER COMPONENTS CONTENT OF L-LYSINE Mobile phase: Methanol and water (31:19) Adjust with glacial acetic acid to a pH of 3.0. Standard stock solution: 0.36 mg/mL of L-lysine Standard solution: Transfer 2.0 mL of Standard stock solution in a glass-stoppered 10-mL volumetric flask, add 2.0 mL of a 1.4% solution of tris(hydroxymethyl)aminomethane and 3.0 mL of a 1.5% solution of 1-fluoro-2,4dinitrobenzene in dehydrated alcohol, and insert the stopper tightly. Heat at 50 in a water bath for 30 min. Remove the flask from the water bath, allow to cool, and dilute with methanol to volume. Sample stock solution: 1.5 mg/mL of Ceforanide for Injection Sample solution: 2.0 mL of Sample stock solution in a glassstoppered 10-mL volumetric flask. Add 2.0 mL of a 1.4% solution of tris(hydroxymethyl)aminomethane and 3.0 mL of a 1.5% solution of 1-fluoro-2,4-dinitrobenzene in dehydrated alcohol, and insert the stopper tightly. Heat at 50 in a water bath for 30 min. Remove the flask from the water bath, allow to cool, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5- to 10-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 4.5 between the derivatized L-lysine peak and the 1-fluoro-2,4-dinitrobenzene peak. Capacity factor, k: 46 Column efficiency: NLT 1500 theoretical plates from the derivatized L-lysine peak Tailing factor: NMT 1.3 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of of L-lysine in the Ceforanide for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of L-lysine in the Standard stock solution (g/mL) = nominal concentration of ceforanide in the CU Sample solution (g/mL) Acceptance criteria: 9001050 g/mg PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: requirements. It meets the rU rS CS
SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.25 USP Endotoxin Unit/mg of ceforanide STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration, except to constitute each container with 3 mL of Fluid A for each g of ceforanide contained therein, and to rinse the membrane with three 100-mL portions of Fluid D and one 100-mL portion of Fluid A. PH 791: 5.58.5, when constituted as directed in the labeling
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USP 32
Standard solution: 0.8 mg/mL of USP Cefotaxime Sodium RS in Solution B [NOTEUse this solution promptly. It may be used within 24 h if stored in a refrigerator.] System suitability solution: Mix 1 mL of Standard solution, 7.0 mL of water, and 2.0 mL of methanol. Add 25 mg of sodium carbonate, mix, and allow to stand at room temperature for 10 min, with occasional swirling. Add 3 drops of glacial acetic acid, and 1 mL of Standard solution. Quantitative limit stock solution: 1.0 mL of Standard solution diluted with Solution B to 50.0 mL Quantitative limit solution: Quantitative limit stock solution diluted with Solution B to 10.0 mL Sample solution: 0.8 mg/mL of Cefotaxime Sodium in Solution B [NOTEUse this solution promptly. It may be used within 24 h if stored in a refrigerator.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 3.9-mm 15-cm; 5-m packing L1 Temperature: 30 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution, System suitability solution, and Quantitative limit solution [NOTERetention times are 3.5 min for desacetylcefotaxime, System suitability solution; 14 min for cefotaxime, System suitability solution; and 1215 min for the main cefotaxime peak, Standard solution.] Suitability requirements [NOTEThe cefotaxime peak response from the Quantitative limit solution is 0.18%0.22% of the cefotaxime peak response from the Standard solution.] Resolution: NLT 20 between the two peaks of desacetylcefotaxime and cefotaxime, System suitability solution Tailing factor: NMT 2, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N5O7S2 in the portion of Cefotaxime Sodium taken: Result = (rU/rS) (CS/CU) Mr1/Mr2 100 = peak response of cefotaxime of the Sample solution = peak response of cefotaxime of the Standard rS solution = concentration of USP Cefotaxime Sodium RS in CS the Standard solution (mg/mL) = nominal concentration of Cefotaxime Sodium in CU the Sample solution (mg/mL) = molecular weight of cefotaxime, 455.47 Mr1 = molecular weight of cefotaxime sodium, 477.45 Mr2 Acceptance criteria: 916964 g/mg rU IMPURITIES Organic Impurities PROCEDURE Sample: Sample solution Calculate the percentage of each impurity: Result = rU/(rT + rC) 100
WATER DETERMINATION, Method I 921: NMT 3.0% PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. OTHER REQUIREMENTS: It meets the requirements under Injections 1, Labels and Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Ceforanide RS USP Endotoxin RS
Cefotaxime Sodium
(Comment on this Monograph)id=m14080=Cefotaxime Sodium=Ca-Chl-Monos.pdf)
477.45 C16H16N5NaO7S2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3(acetyloxy)methyl-7-[[(2-amino-4thiazolyl)(methoxyimino)acetyl]amino]-8-oxo, monosodium salt, [6R-[6,7 (Z)]]-; Sodium (6R,7R)-7-[2-(2-amino-4-thiazolyl)glyoxylamido]-3(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylate 72-(Z)-(O-methyloxime), acetate (ester) [64485-93-4]. DEFINITION Cefotaxime Sodium contains the equivalent of NLT 916 g and NMT 964 g of cefotaxime (C16H17N5O7S2)/mg, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the cefotaxime major peak from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets the requirements. ASSAY PROCEDURE Solution A: 7.1 mg/mL of anhydrous dibasic sodium phosphate in water Adjust with phosphoric acid to a pH of 6.25. Solution B: Methanol and Solution A (7:43) Pass through a filter having a porosity of 0.5 m or less before use. Solution C: Methanol and Solution A (2:3) Pass through a filter having a porosity of 0.5 m or less, before use. Mobile phase: Equilibrate the system with 100% Solution B. Increase the proportion of Solution C linearly from 0%20% at a rate of 10% per min 7 min after injection of the solution under test, and maintain at that composition for 7 min. Increase the proportion of Solution C linearly at a rate of 2.7% /min until the proportion of Solution C is 100%, and hold at that composition for 5 min. Increase the proportion of Solution B linearly to 100% at a rate of 20%/min.
USP 32
rU = peak area of a given impurity = sum of all of the impurity peak areas rT = peak area for the main cefotaxime peak rC [NOTEDisregard any impurity peak that is less than 0.1%.] Acceptance criteria Individual impurities: NMT 1.0% Total impurities: NMT 3.0% SPECIFIC TESTS CLARITY AND COLOR OF SOLUTION: 2.5 g in 25 mL of carbon dioxidefree water Mix, and examine immediately: the solution is clear. Measure the absorbance of this solution at 430 nm in a 1cm cell, using carbon dioxidefree water as the blank: its absorbance is NMT 0.20. Transfer 10 mL of the solution to a glass test tube, add 1 mL of glacial acetic acid, mix, and examine immediately: the solution is clear. OPTICAL ROTATION, Specific Rotation 781S: +58 to +64 Sample solution: 10 mg/mL PH 791: 4.56.5, in a solution (1 in 10) LOSS ON DRYING 731: Dry it at 100105 for 3 h: it loses NMT 3.0% of its weight. STERILITY TESTS 71: Where the label states that Cefotaxime Sodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefotaxime Sodium is sterile or it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.2 Endotoxin Unit/mg of cefotaxime. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefotaxime Sodium RS USP Endotoxin RS
Official Monographs / Cefotaxime 147

Solution C: Methanol and Solution A (2:3) Pass through a filter having a porosity of 0.5 m or less. Mobile phase: Equilibrate the system with 100% Solution B. Increase the proportion of Solution C 7 min after injection of the solution under test, linearly from 0% to 20% at a rate of 10% per min and maintain at that composition for 7 min. Increase the proportion of Solution C linearly at a rate of 2.7% per min until the proportion of Solution C is 100%, and hold at that composition for 5 min. Increase the proportion of Solution B linearly to 100% at a rate of 20% per min. Standard solution: 0.8 mg/mL of USP Cefotaxime Sodium RS in Solution B [NOTEUse this solution promptly. It may be used within 24 h if stored in a refrigerator.] System suitability solution: Mix 1 mL of Standard solution, 7.0 mL of water, and 2.0 mL of methanol. Add 25 mg of sodium carbonate, and allow to stand at room temperature for 10 min, with occasional swirling. Add 3 drops of glacial acetic acid and 1 mL of Standard solution. Quantitative limit stock solution: 1.0 mL of Standard solution diluted with Solution B to 50.0 mL Quantitative limit solution: 1.0 mL of Quantitative limit stock solution diluted with Solution B to 10.0 mL Sample solution: Allow one container of Injection to thaw. Transfer a volume of the Injection, equivalent to 80 mg of C16H17N5O7S2 to a 100-mL volumetric flask, and dilute with Solution B to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 3.9-mm 15-cm; 5-m packing L1 Temperature: 30 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution, Standard solution and Quantitative limit solution [NOTEThe Retention times are: 3.5 min for desacetylcefotaxime and 14 min for cefotaxime, System suitability solution; and 1215 min for the main cefotaxime peak, Standard solution] [NOTEThe response of the cefotaxime peak from the Quantitative limit solution is between 0.18%0.22% of the response of the cefotaxime peak from the Standard solution.] Suitability requirements Resolution: NLT 20 between the two peaks of desacetylcefotaxime and cefotaxime, System suitability solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N5O7S2 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) Mr1/Mr2 100 rU rS CS CU Mr1 Mr2 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cefotaxime Sodium RS in the Standard solution (mg/mL) = nominal concentration of cefotaxime sodium in the Sample solution (mg/mL) = molecular weight of cefotaxime, 455.47 = molecular weight of cefotaxime sodium, 477.45
Cefotaxime Injection
(Comment on this Monograph)id=m14084=Cefotaxime Injection=Ca-Chl-Monos.pdf) DEFINITION Cefotaxime Injection is a sterile solution of Cefotaxime Sodium in Water for Injection. It contains one or more suitable buffers, and it may contain Dextrose or Sodium Chloride as a tonicityadjusting agent. It contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of cefotaxime (C16H17N5O7S2). IDENTIFICATION The retention time of the major peak for cefotaxime in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 7.1 mg/mL of anhydrous dibasic sodium phosphate in water Adjust with phosphoric acid to a pH of 6.25 Solution B: Methanol and Solution A (7:43) Pass through a filter having a porosity of 0.5 m or less.
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90.0%110.0%
USP 32
Solution B: Methanol and Solution A (7:43) Pass through a filter having a porosity of 0.5 m or less, before use. Solution C: Methanol and Solution A (2:3) Pass through a filter having a porosity of 0.5 m or less, before use. Mobile phase: Equilibrate the system with 100% Solution B. Increase the proportion of Solution C linearly from 0%20% at a rate of 10% per min 7 min after injection of the solution under test, and maintain at that composition for 7 min. Increase the proportion of Solution C linearly at a rate of 2.7% /min until the proportion of Solution C is 100%, and hold at that composition for 5 min. Increase the proportion of Solution B linearly to 100% at a rate of 20%/min. Standard solution: 0.8 mg/mL of USP Cefotaxime Sodium RS in Solution B [NOTEUse this solution promptly. It may be used within 24 h if stored in a refrigerator.] System suitability solution: Mix 1 mL of Standard solution, 7.0 mL of water, and 2.0 mL of methanol. Add 25 mg of sodium carbonate, mix, and allow to stand at room temperature for 10 min, with occasional swirling. Add 3 drops of glacial acetic acid and 1 mL of Standard solution. Quantitative limit stock solution: 1.0 mL of Standard solution diluted with Solution B to 50.0 mL. Quantitative limit solution: 1.0 mL of Quantitative limit stock solution diluted with Solution B to 10.0 mL. Sample solution A (for use where the Weight Variation test is to be performed): 0.8 mg/mL of Cefotaxime for Injection in Solution A [NOTEUse this solution promptly. It may be used within 24 h if stored in a refrigerator.] Sample solution B (for use in assaying vials and infusion bottles packaged for dispensing): Constitute one container of Cefotaxime for Injection with the smallest volume of diluent specified in the labeling. Invert the container, and withdraw all of the withdrawable contents of the container with a hypodermic needle and syringe. Transfer the contents of the syringe to a 100-mL volumetric flask, dilute with Solution B to volume, and mix. [NOTEDo not rinse the syringe or container.] Dilute a measured volume of this solution quantitatively with Solution B to obtain a solution having a concentration of 0.8 mg of cefotaxime per mL. [NOTEUse this solution promptly. It may be used within 24 h if stored in a refrigerator.] Sample solution C (for use in assaying piggyback infusion bottles): Constitute one container of Cefotaxime for Injection with the smallest volume of diluent recommended in the labeling, using the directions specified in the labeling. Proceed as directed for Sample solution B beginning with Invert the container. Sample solution D (for use in assaying pharmacy bulk packages where the label states the quantity of cefotaxime in a given volume of constituted solution): Constitute one container of Cefotaxime for Injection with the volume of diluent, specified in the labeling. With a hypodermic needle and syringe, withdraw an accurately measured portion of the resultant solution, equivalent to about 1000 mg of cefotaxime, to a 100-mL volumetric flask, dilute with Solution B to volume, and mix. [NOTEDo not rinse the syringe or container.] Dilute volume of this solution quantitatively with Solution B to obtain a solution having a concentration of 0.8 mg of cefotaxime per mL. [NOTEUse this solution promptly. It may be used within 24 h if stored in a refrigerator.] Chromatographic system (See Chromatography 621, System Suitability.)
IMPURITIES Organic Impurities PROCEDURE Analysis: Using the chromatogram of the Sample solution obtained in the Assay, calculate the percentage of each impurity: Result = rU/(rT + rC) 100 = peak area of a given impurity = sum of all of the impurity peak areas = peak area for the main cefotaxime peak [NOTEDisregard any impurity peak that is less than 0.1%.] Acceptance criteria Individual impurities: NMT 6.0% of any individual impurity is found. Total impurities: NMT 10.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin Unit/mg of cefotaxime STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 5.07.5 PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, as described under Injections 1. Maintain in the frozen state. LABELING: It meets the requirements under Injections 1, Labeling. The label states that it is to be thawed just prior to use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cefotaxime Sodium RS USP Endotoxin RS rU rT rC
Cefotaxime for Injection

(Comment on this Monograph)id=m14087=Cefotaxime for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefotaxime for Injection contains an amount of Cefotaxime Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of cefotaxime (C16H17N5O7S2). IDENTIFICATION Where the Label Indicates That There Are No Added Substances A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets the requirements. Where the Label Indicates That There Are Added Substances C. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 7.1 mg/mL of anhydrous dibasic sodium phosphate in water Adjust with phosphoric acid to a pH of 6.25.
USP 32
Mode: LC Detector: UV 235 nm Column: 3.9-mm 15-cm; 5-m packing L1 Temperature: 30 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard solution, System suitability solution, and Quantitative limit solution[NOTEThe Retention times are: 3.5 min for desacetylcefotaxime and 14 min for cefotaxime, System suitability solution; for the main cefotaxime peak, 1215 min, Standard solution ] [NOTEThe response of the cefotaxime peak from Quantitative limit solution is between 0.18%0.22% of the response of the cefotaxime peak of the Standard solution.] Suitability requirements Resolution: NLT 20 between the two peaks of desacetylcefotaxime and cefotaxime, System suitability solution Tailing factor: NMT 2, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Sample solution A, or Sample solution B, or Sample solution C, or Sample solution D, and Standard solution Calculate the percentage of C16H17N5O7S2 in each mg of the Cefotaxime for Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cefotaxime Sodium RS in the Standard solution (mg/mL) = nominal concentration of cefotaxime sodium in CU the Sample solution (mg/mL) = molecular weight of cefotaxime, 455.47 Mr1 = molecular weight of cefotaxime sodium, 477.45 Mr2 Acceptance criteria: 90.0%115.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 Analysis: Perform the Assay on individual containers using Sample solution B, Sample solution C, or Sample solution D, as appropriate. IMPURITIES Organic Impurities PROCEDURE Using the chromatogram of the Sample solution obtained in the Assay, calculate the percentage of each impurity: Result = rU/(rT + rc) 100 rU rT rc = peak area of a given impurity = sum of all of the impurity peak area responses = peak area for the main cefotaxime peak [NOTEDisregard any impurity peak that is less than 0.1%.] Acceptance criteria Individual impurities: NMT 6.0% of any individual impurity is found. Total impurities: NMT 10.0%
Official Monographs / Cefotetan 149

PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. PH 791: 4.56.5, in a solution (1 in 10) LOSS ON DRYING 731: Dry it at 100105 for 3 h: it loses NMT 3.0% of its weight. OTHER REQUIREMENTS: It meets the requirements under Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefotaxime Sodium RS USP Endotoxin RS
Cefotetan
(Comment on this Monograph)id=m14093=Cefotetan=Ca-ChlMonos.pdf)
575.62 C17H17N7O8S4 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[4-(2amino-1-carboxy-2-oxoethylidene)-1,3-dithietan-2yl]carbonyl]amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, [6R-(6,7)]-; (6R,7S)-4-[[2-Carboxy-7-methoxy-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-7yl]carbamoyl]-1,3-dithietane-2,-malonamic acid; (6R,7S)-7-[4-(Carbamoylcarboxymethylene)-1,3-dithiethane-2carboxamido]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylic acid [69712-56-7]. DEFINITION Cefotetan contains NLT 950 g and NMT 1030 g of cefotetan (C17H17N7O8S4)/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution as directed in the Assay. ASSAY PROCEDURE [NOTEProtect the Standard solution, the System suitability solution, and the Sample solution from light, and use within 2 h.] Mobile phase: Methanol, acetonitrile, glacial acetic acid, and 0.1 M phosphoric acid (105:105:100:1700) Standard solution: 20 mg of USP Cefotetan RS in a 100mL volumetric flask, add 5 mL of methanol, swirl for several min, add 5 mL of acetonitrile, and swirl until dissolved. Dilute with water to volume. System suitability solution: 10 mL of Standard solution in a glass-stoppered flask containing a few mg of magnesium carbonate. Sonicate for 10 min. If the solution is not turbid, add a few more mg of magnesium carbonate, and repeat the sonication. Filter the turbid solution through a filter of 0.5m or finer porosity. Use the clear filtrate.
SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: NMT 0.2 USP Endotoxin Unit/mg of cefotaxime STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration.
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Cefotetan / Official Monographs
USP 32
Sample solution: 20 mg of Cefotetan in a 100-mL volumetric flask Add 5 mL of methanol, swirl for several min, add 5 mL of acetonitrile, and swirl until dissolved. Dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times are 0.75 for cefotetan and 1.0 for cefotetan tautomer, System suitability solution.] Suitability requirements Resolution: NLT 2.0 between the cefotetan peak and the cefotetan tautomer peak, System suitability solution Column efficiency: NLT 1500 theoretical plates, Standard solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C17H17N7O8S4 in each mg: Result = (rU/rS) (CS/CU) = peak response of cefotetan from the Sample solution = peak response of cefotetan from the Standard rS solution = concentration of USP Cefotetan RS in the CS Standard solution (mg/mL) = nominal concentration of cefotetan in the CU Sample solution (mg/mL) Acceptance criteria: 9501030 g/mg rU SPECIFIC TESTS STERILITY TESTS 71: Where the label states that Cefotetan is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration, except to use Fluid A to each 1000 mL of which has been added 10 g of sodium bicarbonate before sterilization. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefotetan is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.17 USP Endotoxin Unit/mg of cefotetan. WATER DETERMINATION, Method I 921: NMT 2.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefotetan RS USP Endotoxin RS
Cefotetan Injection
(Comment on this Monograph)id=m14094=Cefotetan Injection=Ca-Chl-Monos.pdf) DEFINITION Cefotetan Injection is a sterile isoosmotic solution of Cefotetan and Sodium Bicarbonate in Water for Injection. It contains one or more buffer substances and a tonicity-adjusting agent. It contains NLT 90.0% and NMT 120.0% of the labeled amount of cefotetan (C17H17N7O8S4). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that in the Standard solution obtained as directed in the Assay. ASSAY [NOTEProtect the Standard solution, the System suitability solution, and the Sample solution from light, and use within 2 h.] PROCEDURE Mobile phase: Methanol, acetonitrile, glacial acetic acid, and 0.1 M phosphoric acid (105:105:100:1700) Standard solution: 20 mg of USP Cefotetan RS in a 100mL volumetric flask Add 5 mL of methanol, swirl for several min, add 5 mL of acetonitrile, and swirl until dissolved. Dilute with water to volume. System suitability solution: 10 mL of Standard solution in a glass-stoppered flask containing a few mg of magnesium carbonate Sonicate for 10 min. If the solution is not turbid, add a few more mg of magnesium carbonate, and repeat the sonication. Filter the turbid solution through a filter of 0.5 m or finer porosity. Use the clear filtrate. Sample solution: Allow the contents of a container of Injection to thaw, and mix the resultant solution. Transfer a volume of this solution, equivalent to about 40 mg of cefotetan, to a 200-mL volumetric flask, and dilute with Mobile phase to volume. [NOTEUse the sample solution within 10 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for cefotetan and cefotetan tautomer, System suitability solution, are about 0.75 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the cefotetan peak and the cefotetan tautomer peak, System suitability solution Column efficiency: NLT 1500 theoretical plates, Standard solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution
USP 32
Analysis Sample: Standard solution and Sample solution Calculate the percentage of C17H17N7O8S4 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cefotetan RS in the Standard solution (mg/mL) = nominal concentration of cefotetan in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%120.0% rU rS CS SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.17 USP Endotoxin Unit/mg of cefotetan STERILITY TESTS 71: Meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 4.06.5 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: Meets the requirements for Injections 1, Labeling. The label states that it is to be thawed just prior to use, describes the conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cefotetan RS USP Endotoxin RS
Official Monographs / Cefotetan 151

Solution A to obtain a solution containing the equivalent of 200 g of cefotetan/mL. Sample solution B (where the label states the quantity of cefotetan in a given volume of constituted solution): Constitute Cefotetan for Injection as directed in the labeling. Dilute a volume of the constituted solution quantitatively with Solution A to obtain a solution containing the equivalent of 200 g of cefotetan/mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cefotetan and cefotetan tautomer are 0.75 and 1.0, respectively, in System suitability solution.] Suitability requirements Resolution: NLT 2.0 between the cefotetan peak and the cefotetan tautomer peak, System suitability solution Column efficiency: NLT 1500 theoretical plates, Standard solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H17N7O8S4 withdrawn from the container, or in the portion of solution taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cefotetan RS in the Standard solution (g/mL) CU = nominal concentration of cefotetan in Sample solution A or Sample solution B (g/mL) Acceptance criteria: 90.0%120.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: requirements Meets the
Cefotetan for Injection

(Comment on this Monograph)id=m14095=Cefotetan for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefotetan for Injection contains an amount of Cefotetan Disodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cefotetan (C17H17N7O8S4). ASSAY PROCEDURE [NOTEProtect the Standard solution, the System suitability solution, and the Sample solution from light, and use within 2 h.] Solution A: Methanol, acetonitrile, and water (1:1:18) Mobile phase: Methanol, acetonitrile, glacial acetic acid, and 0.1 M phosphoric acid (105:105:100:1700) Standard solution: 20 mg of USP Cefotetan RS in a 100mL volumetric flask Add 5 mL of methanol, swirl for several minutes, add 5 mL of acetonitrile, and swirl until dissolved. Dilute with water to volume. System suitability solution: 10 mL of Standard solution in a glass-stoppered flask containing a few mg of magnesium carbonate Sonicate for 10 min. If the solution is not turbid, add a few more mg of magnesium carbonate, and repeat the sonication. Filter the turbid solution through a filter of 0.5 m or finer porosity. Use the clear filtrate. Sample solution A (where the package is represented as being in a single-dose container): Constitute Cefotetan for Injection as directed in the labeling. Withdraw all of the withdrawable contents, and quantitatively dilute with
SPECIFIC TESTS INJECTIONS, Constituted Solutions 1: Meets the requirements BACTERIAL ENDOTOXINS TEST 85 : NMT 0.17 USP Endotoxin Unit/mg of cefotetan STERILITY TESTS 71: Meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 4.06.5, in a solution (1 in 10) Change to read: WATER DETERMINATION, Method Ic 921: NMT 2.8%3 OTHER REQUIREMENTS: It meets the requirements of the Identification tests under Cefotetan Disodium and meets the requirements under Injections 1, Labels and Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids.
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Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for cefotetan and cefotetan tautomer are 0.75 and 1.0, respectively, from the System suitability solution.] Suitability requirements Resolution: NLT 2.0 between the cefotetan peak and the cefotetan tautomer peak, System suitability solution Column efficiency: NLT 1500 theoretical plates, Standard solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Sample: Standard solution and Sample solution Calculate the quantity, in g, of C17H17N7O8S4 per mg in the portion of Cefotetan Disodium taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) x P = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cefotetan RS in the Standard solution (g/mL) CU = nominal concentration of cefotetan disodium in the Sample solution (g/mL) = molecular weight of cefotetan, 575.62 Mr1 = molecular weight of cefotetan disodium, 619.59 Mr2 P = stated content of cefotetan in USP Cefotetan RS (mg/mg) Acceptance criteria: 830970 g/mg rU rS CS SPECIFIC TESTS PH 791: 4.06.5, in a solution (1 in 10) Change to read: WATER DETERMINATION, Method Ic 921: NMT 2.5%3 STERILITY TESTS 71: Where the label states that Cefotetan Disodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefotetan Disodium is sterile or it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.17 USP Endotoxin Unit/mg of cefotetan. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefotetan RS USP Endotoxin RS
USP REFERENCE STANDARDS 11 USP Cefotetan RS USP Endotoxin RS
Cefotetan Disodium
(Comment on this Monograph)id=m14096=Cefotetan Disodium=Ca-Chl-Monos.pdf)
619.59 C17H15N7Na2O8S4 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[4-(2amino-1-carboxy-2-oxoethylidene)-1,3-dithietan-2-yl]carbonyl] amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-, disodium salt, [6R-(6,7)]-; (6R,7S)-4-[[2-Carboxy-7-methoxy-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-7yl]carbamoyl]-1,3-dithietane-2,-malonamic acid, disodium salt; (6R,7S)-7-[4-(Carbamoylcarboxymethylene)-1,3-dithietane-2carboxamido]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylic acid, disodium salt [74356-00-6]. DEFINITION Cefotetan Disodium contains the equivalent of NLT 830 g and NMT 970 g of cefotetan (C17H17N7O8S4) per mg, calculated on the anhydrous basis. IDENTIFICATION A. The retention time of the major peak from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements ASSAY PROCEDURE [NOTEProtect the Standard solution, the System suitability solution, and the Sample solution from light, and use within 2 h.] Mobile phase: Methanol, acetonitrile, glacial acetic acid, and 0.1 M phosphoric acid (105:105:100:1700) Standard solution: 20 mg of USP Cefotetan RS in a 100mL volumetric flask Add 5 mL of methanol, swirl for several min, add 5 mL of acetonitrile, and swirl until dissolved. Dilute with water to volume. System suitability solution: 10 mL of Standard solution in a glass-stoppered flask containing a few mg of magnesium carbonate Sonicate for 10 min. If the solution is not turbid, add a few more mg of magnesium carbonate, and repeat the sonication. Filter the turbid solution through a filter of 0.5 m or finer porosity. Use the clear filtrate. Sample solution: 40 mg of Cefotetan Disodium in a 200mL volumetric flask Add 10 mL of methanol, swirl for several minutes, add 10 mL of acetonitrile, and swirl until dissolved. Dilute with water to volume.
USP 32
Official Monographs / Cefotiam 153

Column efficiency: From the cefotiam peak, NLT 1985 theoretical plates, Standard solution, when calculated as: Result = 5.545(tr/Wh/2)2 Wh/2 = width of peak at half-height Tailing factor: NMT 1.8, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C18H23N9O4S3 in each mg of the Cefotiam Hydrochloride: Result = (rU/rS) (CS/CU) = peak response of the Sample solution = peak response of the Standard solution = concentration of cefotiam in the Standard solution (g/mL) = concentration of Cefotiam Hydrochloride in the CU Sample solution (mg/mL) Acceptance criteria: 790925 g/mg rU rS CS SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PYROGEN TEST 151: Where the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements of the test, the test dose being 1.0 mL/kg of a solution in pyrogen-free sodium carbonate solution containing 40 mg/mL (prepared by dissolving 25.6 g of sodium carbonate, previously heated at 170 for NLT 4 h, in 1000 mL of Sterile Water for Injection). STERILITY TESTS 71: Where the label states that it is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. WATER DETERMINATION, Method I 921: NMT 7.0%, the Sample solution being prepared as directed for a hygroscopic specimen, except to use a mixture of 20 mL of formamide (previously dried over anhydrous sodium sulfate for 24 h) and methanol (2:1), instead of methanol, to dissolve the specimen, and to determine the water content of the formamide and methanol mixture ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefotiam Hydrochloride RS
Cefotiam Hydrochloride
(Comment on this Monograph)id=m14098=Cefotiam Hydrochloride=Ca-Chl-Monos.pdf)
C18H23N9O4S3 2HCl 598.56 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7[[-(2amino-4-thiazolyl)acetyl]-amino]-3-[[[1-[2-(dimethylamino) ethyl]-1H-tetrazol-5-yl]-thio]methyl]-8-oxo, hydrochloride, (6R-trans)-; (6R,7R)-7-[2-(2-Amino-4-thiazolyl)acetamido]-3-[[[1-[2(dimethylamino)ethyl]-1H-tetrazol-5-yl]thio]methyl]-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid dihydrochloride; 7(R)-[2-(2-Amino-4-thiazolyl)acetamido]-3-[[[1-[2dimethylamino)ethyl]-1H-tetrazol-5-yl]thio]methyl]-3cephem-4-carboxylic acid dihydrochloride [66309-69-1]. DEFINITION Cefotiam Hydrochloride contains the equivalent of NLT 790 g and NMT 925 g of cefotiam (C18H23N9O4S3)/mg, calculated on the anhydrous basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Solution: 20 g/mL in water B. The retention time of the cefotiam peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 13.1 g of ammonium sulfate in 850 mL of water Adjust with 2 N ammonium hydroxide to a pH of 6.5 0.1, and add 150 mL of acetonitrile. Pass through a suitable filter of 0.5 m or finer porosity. System suitability stock solution: 1 mg/mL of USP Cefotiam Hydrochloride RS [NOTEHeat this solution at 95 for 3 min, and cool.] System suitability solution: 1 mL of System suitability stock solution diluted with Mobile phase to 100 mL Standard stock solution: 1 mg/mL of cefotiam from USP Cefotiam Hydrochloride RS Standard solution: 50 g/mL cefotiam from Standard stock solution diluted with Mobile phase [NOTEUse this solution without delay.] Sample stock solution: 1.2 mg/mL of Cefotiam Hydrochloride Sample solution: 60 g/mL from Sample stock solution diluted with Mobile phase [NOTEUse this solution without delay.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 25-cm column; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for de-tetrazol-cefotiam and cefotiam are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 4.0 between the de-tetrazol-cefotiam peak and the cefotiam peak, System suitability solution
Cefotiam for Injection

(Comment on this Monograph)id=m14099=Cefotiam for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefotiam for Injection contains an amount of Cefotiam Hydrochloride equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cefotiam (C18H23N9O4S3). It may contain Sodium Carbonate. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Solution: 20 g/mL B. The retention time of the major peak for cefotiam in the Sample solution corresponds to that in the Standard solution as obtained in the Assay.
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rU rS CS
USP 32
= peak response from the Sample solution = peak response from the Standard solution = concentration of cefotiam in the Standard solution (mg/mL) = nominal concentration of cefotiam in Sample CU solution A or Sample solution B (g/mL) Acceptance criteria: 90.0%120.0% SPECIFIC TESTS PYROGEN TEST 151: Meets the requirements of the test; the dose being 1.0 mL/kg of a solution prepared by diluting Cefotiam for Injection with Sterile Water for Injection to a concentration of 40 mg/mL of cefotiam STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 5.77.2, in a solution of 100 mg of cefotiam/mL LOSS ON DRYING 731: Dry 100 mg in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 6.0% of its weight. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefotiam Hydrochloride RS
ASSAY PROCEDURE Mobile phase: 13.1 g of ammonium sulfate in 850 mL of water Adjust with 2 N ammonium hydroxide to a pH of 6.5 0.1, and add 150 mL of acetonitrile. Pass through a suitable filter of 0.5 m or finer porosity. System suitability stock solution: 1 mg/mL of USP Cefotiam Hydrochloride RS [NOTEHeat this solution at 95 for 3 min and cool.] System suitability solution: Mobile phase and System suitability stock solution (99:1) Standard stock solution: 1 mg/mL of USP Cefotiam Hydrochloride RS Standard solution: 50 g/mL from the Standard stock solution in Mobile phase [NOTEUse this solution without delay.] Sample stock solution A (where it is represented as being in a single-dose container): Constitute a container of Cefotiam for Injection in a volume of water, measured corresponding to the volume of diluent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute quantitatively with water to obtain a solution containing the equivalent of 1 mg of cefotiam/mL. Sample solution A: 50 g/mL from Sample stock solution A in Mobile phase [NOTEUse this solution without delay.] Sample stock solution B (where the label states the quantity of cefotiam in a given volume of constituted solution): Constitute a container of Cefotiam for Injection in a volume of water, measured and equivalent to the volume of diluent specified in the labeling. Dilute a measured volume of the constituted solution quantitatively with water to obtain a solution containing 1 mg of cefotiam/mL. Sample solution B: 50 g/mL from Sample stock solution B in Mobile phase [NOTEUse this solution without delay.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 25-cm column; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for de-tetrazol-cefotiam and cefotiam are about 0.6 and 1.0, respectively, System suitability solution.] Suitability requirements Resolution: NLT 4.0 between the de-tetrazol-cefotiam peak and the cefotiam peak, System suitability solution Column efficiency: NLT 1985 theoretical plates from the cefotiam peak of the Standard solution when calculated by the following formula: 5.545(TR/WH/2)2 Tailing factor: NMT 1.8, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Sample: Sample solution A or Sample solution B, and Standard solution Calculate the percentage of C18H23N9O4S3 withdrawn from the container, or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100
Cefoxitin Sodium
(Comment on this Monograph)id=m14104=Cefoxitin Sodium=Ca-Chl-Monos.pdf)
C16H16N3NaO7S2 449.44 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[[(aminocarbonyl)oxy]methyl]-7-methoxy-8-oxo-7[(2-thienylacetyl)-amino]-, sodium salt (6R-cis)-; Sodium (6R,7S)-3-(hydroxymethyl)-7-methoxy-8-oxo-7-[2-(2thienyl)acetamido]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylate carbamate (ester) [33564-30-6; 35607-66-0]. DEFINITION Cefoxitin Sodium contains the equivalent of NLT 927 g/mg and NMT 970 g/mg of cefoxitin (C16H17N3O7S2), corresponding to NLT 97.5% and NMT 102.0% of cefoxitin sodium (C16H16N3NaO7S2), calculated on the anhydrous and acetone- and methanol-free basis. IDENTIFICATION A. The retention time of the major peak for cefoxitin in the Sample solution corresponds to that of the Standard solution, obtained as directed in the Assay. B. ULTRAVIOLET ABSORPTION 197U Buffer solution: 1.0 mg/mL of monobasic potassium phosphate and 1.8 mg/mL of anhydrous dibasic sodium phosphate
USP 32
Sample solution: 20 g/mL in Buffer solution C. IDENTIFICATION TESTSGENERAL, Sodium 191 Sample solution: 50 mg/mL Acceptance criteria: Meets the requirements ASSAY PROCEDURE Mobile phase: Acetonitrile, water, and glacial acetic acid (16:84:1) Filter through a membrane filter of 1-m or finer porosity. Buffer solution: 1.0 g of monobasic potassium phosphate and 1.8 g of dibasic sodium phosphate in 900 mL of water Adjust with phosphoric acid or 10 N sodium hydroxide to a pH of 7.1 0.1, and dilute with water to make 1000 mL. Pass through a membrane filter of 1-m or finer porosity. Standard solution: 0.3 mg/mL of USP Cefoxitin RS in Buffer solution [NOTESonicate, if necessary, to dissolve the specimen and use this solution within 5 h.] Sample solution: 0.3 mg/mL of Cefoxitin Sodium in Buffer solution [NOTEUse this solution within 5 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 5- to 10-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2800 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.0% Analysis Samples: Sample solution and Standard solution Calculate the concentration, in g/mg, of C16H17N3O7S2 in the portion of Cefoxitin Sodium taken: Result = (rU/rS) CS/CU) P = peak response from the Sample solution = peak response from the Standard solution = concentration of cefoxitin in the Standard solution (mg/mL) = nominal concentration of Cefoxitin Sodium CU taken to prepare the Sample solution (mg/mL) P = potency of USP Cefoxitin RS (g/mg) Acceptance criteria: 927970 g/mg rU rS CS IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities LIMIT OF ACETONE AND METHANOL Standard stock solution A: 0.5% acetone in water Standard stock solution B: 0.5% methanol in water Standard solution: 0.050% acetone and 0.005% methanol from Standard stock solution A and Standard stock solution B diluted with water Sample solution: To a 50-mL volumetric flask, add 5.0 g of Cefoxitin Sodium, and dissolve in and dilute with water to volume. Transfer 3.0 mL of the resulting solution to a 15-mL centrifuge tube, cool in an ice-water bath for 2 min, and add 3.0 mL of 0.24 N hydrochloric acid while swirling vigorously. Centrifuge to obtain a clear solution, and use the supernatant. Chromatographic system (See Chromatography 621, System Suitability.)
Official Monographs / Cefoxitin 155

Mode: GC Detector: Flame ionization Column: 1.8-m 6.3-mm glass column containing support S2, and a precolumn packed with 60- to 80-mesh silane-treated glass beads Temperature Injection port: 100 Columns: 110 Detector: 200 Carrier gas: Nitrogen Flow rate: 50 mL/min Injection size: 2 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 160 and NLT 200 theoretical plates, respectively, for the acetone and methanol peaks Tailing factor: NMT 1.3 and NMT 2.3, respectively, for the acetone and methanol peaks Relative standard deviation: NMT 5.0% Analysis Samples: Sample solution and Standard solution Calculate the percentages of acetone and methanol in the portion of Cefoxitin Sodium taken: Result = (rU/rS) CS/CU) D = peak response of acetone or methanol in the Sample solution = peak response of acetone or methanol in the rS Standard solution = concentration of acetone or methanol in the CS Standard solution (%) = nominal concentration of Cefoxitin Sodium in CU the Sample solution (g/mL) D = density of acetone and methanol at 20 (g/mL), 0.79 Acceptance criteria: NMT 0.7% of acetone and NMT 0.1% of methanol SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +206 to +214, calculated on the anhydrous and acetone- and methanol-free basis Sample solution: 10 mg/mL, in methanol CRYSTALLINITY 695: Meets the requirements PH 791: 4.27.0, in a solution containing 100 mg/mL WATER DETERMINATION, Method I 921: NMT 1.0%, a mixture of ethylene glycol and pyridine (3:1) being used in place of methanol in the titration vessel BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cefoxitin Sodium is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.13 USP Endotoxin Unit/mg of cefoxitin. STERILITY TESTS 71: Where the label states that Cefoxitin Sodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a cold place. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefoxitin RS rU
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STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791: 4.58.0 PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements for Injections 1, Labeling. The label states that it is to be thawed just prior to use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cefoxitin RS USP Endotoxin RS
USP Endotoxin RS
Cefoxitin Injection
(Comment on this Monograph)id=m14106=Cefoxitin Injection=Ca-Chl-Monos.pdf) DEFINITION Cefoxitin Injection is a sterile solution of Cefoxitin Sodium and one or more suitable buffer substances in Water for Injection. It contains Dextrose or Sodium Chloride as a tonicity-adjusting agent. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cefoxitin (C16H17N3O7S2). IDENTIFICATION The retention time of the major peak for cefoxitin in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, glacial acetic acid, and water (16:1:84) Filter through a membrane filter of 1 m or finer porosity. Solution A: Dissolve 1.0 mg/mL of monobasic potassium phosphate and 1.8 mg/mL of dibasic sodium phosphate in water, and adjust with phosphoric acid or 10 N sodium hydroxide to a pH of 7.1 0.1 [NOTEFilter through a membrane filter of 1 m or finer porosity.] Standard solution: 0.3 mg/mL of USP Cefoxitin RS in Solution A [NOTESonicate, if necessary, to dissolve the specimen, and use this solution within 5 h.] Sample solution: Allow one container of Injection to thaw, and mix. Dilute a volume of Injection quantitatively with Solution A to obtain a solution containing 0.3 mg of cefoxitin/mL. [NOTEUse this solution within 5 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 5 to 10-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2800 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O7S2 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of cefoxitin in the Standard solution (mg/mL) = nominal concentration of cefoxitin in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%120.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: Unit/mg of cefoxitin NMT 0.13 USP Endotoxin rU rS CS
Cefoxitin for Injection

(Comment on this Monograph)id=m14107=Cefoxitin for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefoxitin for Injection contains Cefoxitin Sodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cefoxitin (C16H17N3O7S2). IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. ULTRAVIOLET ABSORPTION 197U Solution: 20 g/mL Medium: 1 mg/mL of monobasic potassium phosphate and 1.8 mg/mL of anhydrous dibasic sodium phosphate in water C. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements ASSAY PROCEDURE Mobile phase: Acetonitrile, glacial acetic acid, and water (16:1:84) Filter through a membrane filter of 1-m or finer porosity. Solution A: 1.0 mg/mL of monobasic potassium phosphate and 1.8 mg/mL of dibasic sodium phosphate in water Adjust with phosphoric acid or 10 N sodium hydroxide to a pH of 7.1 0.1. Filter through a membrane filter of 1-m or finer porosity. Standard solution: 0.3 mg/mL of USP Cefoxitin RS in Solution A [NOTESonicate, if necessary, to dissolve the specimen, and use this solution within 5 h.] Sample solution A (where it is represented as being in a single-dose container): Constitute Cefoxitin for Injection in a volume of water, corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute quantitatively with water to obtain a solution having a nominal concentration of 0.3 mg of cefoxitin/mL. [NOTEUse this solution within 5 h.] Sample solution B (where the label states the quantity of cefoxitin in a given volume of constituted solution): Constitute Cefoxitin for Injection in a volume of water, corresponding to the volume of solvent specified in the labeling. Dilute a measured volume of the constituted solution quantitatively with water to obtain a solution
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having a nominal concentration of 0.3 mg of cefoxitin/mL. [NOTEUse this solution within 5 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 5 to 10-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2800 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.0% Analysis Samples: Sample solution A or Sample solution B and Standard Solution Calculate the percentage of C16H17N3O7S2 in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of cefoxitin in the Standard solution (mg/mL) = nominal concentration of cefoxitin in Sample CU solution A or Sample solution B (mg/mL) Acceptance criteria: 90.0%120.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Official Monographs / Cefpiramide 157

USP Endotoxin RS
Cefpiramide
(Comment on this Monograph)id=m14109=Cefpiramide=CaChl-Monos.pdf)
612.64 C25H24N8O7S2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[[[(4-hydroxy-6-methyl-3-pyridinyl)carbonyl]amino](4hydroxyphenyl)acetyl]amino]-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo, [6R-[6,7 (R*)]]-; (6R,7R)-7-[(R)-2-(4-Hydroxy-6-methylnicotinamido)-2-(phydroxyphenyl)acetamido]-3-[[(1-methyl-1H-tetrazol-5yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylic acid [70797-11-4]. DEFINITION Cefpiramide contains NLT 974 g and NMT 1026 g of C25H24N8O7S2/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.36 mg/mL of monobasic potassium phosphate in water Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1 Mobile phase: Tetrahydrofuran, acetonitrile, methanol, and Solution A (3:1:1:45) System suitability solution: 1 mg/mL of USP Cefpiramide RS in 0.01 N sodium hydroxide Heat this solution at 95 for 10 min. Mix 1 mL of this solution with 19 mL of Mobile phase. This solution contains a mixture of cefpiramide and cefpiramide lactone. Standard solution: 0.25 mg/mL of USP Cefpiramide RS in Mobile phase Sample solution: 0.25 mg/mL of Cefpiramide in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: NMT 0.13 USP Endotoxin Unit/mg of cefoxitin STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. PH 791: 4.27.0, in a solution containing 100 mg/mL WATER DETERMINATION, Method I 921: NMT 1.0%, a mixture of ethylene glycol and pyridine (3:1) being used in place of methanol in the titration vessel OTHER REQUIREMENTS: It meets the requirements for Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefoxitin RS
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System suitability: Proceed as directed in the Assay. Analysis Samples: Standard solution and Sample solution 2 Calculate the percentage of 5-mercapto-1-methyltetrazole in the portion of Cefpiramide taken: Result = (rU/rS) (Mr1/Mr2) [5CS (100-w)/100W] 100 = peak area of 5-mercapto-1-methyltetrazole from Sample solution 2 = peak area of 5-mercapto-1-methyltetrazole from rS the Standard solution = molecular weight of 5-mercapto-1Mr1 methyltetrazole, 115.14 = molecular weight of anhydrous sodium 5Mr2 mercapto-1-methyltetrazole, 138.13 = concentration of sodium 5-mercapto-1CS methyltetrazole in the Standard solution (g/mL) w = percentage of water in the sodium 5mercapto-1-methyltetrazole used to prepare the Standard solution. W = weight of Cefpiramide taken to prepare the Sample solution 2 (mg) Calculate the percentage of each other impurity in the portion of Cefpiramide taken: rU Result = (rU/rS) (CS/CU) P F 100 = peak response of each other impurity from Sample solution 2 rS = peak response of cefpiramide from the Standard solution CS = concentration of USP Cefpiramide RS in the Standard solution (g/mL) = concentration of Cefpiramide in Sample solution CU 2 (g/mL) P = designated cefpiramide content of USP Cefpiramide RS (g/mg) F = correction factor, 0.001 mg/g Acceptance criteria Individual impurities: NMT 0.7% of 5-mercapto-1methyltetrazole; NMT 0.7% of any other individual impurity Total impurities: The sum of 5-mercapto-1methyltetrazole and all other impurities is NMT 2.0%. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 100 to 112 Sample solution: 10 mg/mL, in dimethylformamide CRYSTALLINITY 695: Meets the requirements PH 791: 3.05.0, in a suspension (1 in 200) WATER DETERMINATION, Method I 921: NMT 9.0% PYROGEN TEST 151: Where the label states that Cefpiramide is sterile, or it must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements, the test dose being 1.0 mL/kg of a solution prepared by diluting Cefpiramide with Sterile Water for Injection to a concentration of 50 mg/mL of cefpiramide. STERILITY TESTS 71: Where the label states that Cefpiramide is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. rU
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cefpiramide and cefpiramide lactone are about 0.7and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the cefpiramide lactone peak and the cefpiramide peak, System suitability solution Capacity factor, k: Between 2 and 5, Standard solution Column efficiency: NLT 1700 theoretical plates, from the Standard solution when calculated: Result = 5.545(tr/Wh/2)2 Tailing factor: 0.951.4 for the cefpiramide peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C25H24N8O7S2 in each mg of the Cefpiramide taken: Result = (rU/rS) (CS/CU) P = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cefpiramide RS in the Standard solution (mg/mL) = concentration of Cefpiramide taken to prepare CU the Sample solution (mg/mL) P = designated cefpiramide content of USP Cefpiramide RS (g/mg) Acceptance criteria: 9741026 g/mg IMPURITIES Organic Impurities PROCEDURE Solution: 4.08 mg/mL of monobasic potassium phosphate in water, adjust with 1 N sodium hydroxide to a pH of 7.5 0.1 Mobile phase: Methanol and Solution A (1:3) System suitability solution: Proceed as directed in the Assay. Sodium 5-mercapto-1-methyltetrazole standard: Determine the water content of the Sodium 5-mercapto-1methyltetrazole standard (see Water Determination 921). Sample solution 1: Transfer 100 mg of Sodium 5mercapto-1-methyltetrazole standard to a stopper centrifuge tube. Add 10.0 mL of a solution of Nethylmaleimide in methanol (4 in 100), and sonicate for 15 min. Titrate 5.0 mL of this solution as directed in Water Determination 921. Standard stock solution: 0.15 mg/mL of sodium 5mercapto-1-methyltetrazole and 0.25 mg/mL of USP Cefpiramide RS in Solution A Standard solution: 3 g/mL of sodium 5-mercapto-1methyltetrazole and 5 g/mL of USP Cefpiramide RS from the Standard stock solution diluted with Mobile phase Sample solution 2: 0.5 mg/mL of Cefpiramide in Mobile phase Chromatographic system: Proceed as directed in the Assay. rU rS CS
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefpiramide RS
Official Monographs / Cefpodoxime 159

Tailing factor: 0.951.4 for the cefpiramide peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample Solution A or Sample solution B, and Standard Solution Calculate the percentage of C25H24N8O7S2 withdrawn from the container, or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100 = peak area of the Sample solution = peak area of the Standard solution = concentration of USP Cefpiramide RS in the Standard solution (mg/mL) CU = nominal concentration of cefpiramide in Sample solution A or Sample solution B (mg/mL) Acceptance criteria: 90.0%120.0% rU rS CS SPECIFIC TESTS PYROGEN TEST 151: It meets the requirements, the test dose being 1.0 mL/kg of a solution prepared by diluting Cefpiramide for Injection with Sterile Water for Injection to a concentration of 50 mg/mL of cefpiramide. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 6.08.0 Sample solution: Containing the equivalent of 100 mg/mL of cefpiramide WATER DETERMINATION, Method I 921: NMT 3.0% PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefpiramide RS
Cefpiramide for Injection

(Comment on this Monograph)id=m14110=Cefpiramide for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefpiramide for Injection contains NLT 90.0% and NMT 120.0% of the labeled amount of cefpiramide (C25H24N8O7S2). IDENTIFICATION The retention time for the cefpiramide peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.36 mg/mL of monobasic potassium phosphate in water Adjust with 1 N sodium hydroxide to a pH of 6.8 0.1. Mobile phase: Tetrahydrofuran, acetonitrile, methanol, and Solution A (3:1:1:45) System suitability solution: 1 mg/mL of USP Cefpiramide RS in 0.01 N sodium hydroxide Heat this solution at 95 for 10 min. Mix 1 mL of this solution with 19 mL of Mobile phase. This solution contains a mixture of cefpiramide and cefpiramide lactone. Standard solution: 0.25 mg/mL of USP Cefpiramide RS in Mobile phase Sample solution A (where it is represented as being in a single-dose container): Constitute a container of Cefpiramide for Injection in a volume of water corresponding to the volume of diluent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with Mobile phase to obtain a solution containing the nominal equivalent of 0.25 mg/mL of cefpiramide. Sample solution B (where the label states the quantity of cefpiramide in a given volume of constituted solution): Constitute a container of Cefpiramide for Injection in a volume of water equivalent to the volume of diluent specified in the labeling. Dilute the constituted solution with water to obtain a solution nominally containing 0.25 mg/mL of cefpiramide. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cefpiramide and cefpiramide lactone are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the cefpiramide lactone peak and the cefpiramide peak, System suitability solution Capacity factor, k: 25, Standard solution Column efficiency: NLT 1700 theoretical plates, Standard solution, when calculated: Result = 5.545 (tr/Wh/2)2
Cefpodoxime Proxetil
(Comment on this Monograph)id=m14111=Cefpodoxime Proxetil=Ca-Chl-Monos.pdf)
557.60 C21H27N5O9S2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-3(methoxymethyl)-8-oxo-,1-[[(1-methylethoxy)carbonyl]oxy] ethyl ester, [6R-[6,7(Z)]]-; ()-1-Hydroxyethyl(+)-(6R,7R)-7-[2-(2-amino-4-thiazolyl) glyoxylamido]-3-methoxymethyl)-8-oxo-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylate, 72-(Z)-(O-methyloxime), isopropyl carbonate (ester) [87239-81-4]. DEFINITION Cefpodoxime Proxetil contains the equivalent of NLT 690 g/ mg and NMT 804 g/mg of cefpodoxime (C15H17N5O6S2), calculated on the anhydrous basis.
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Acceptance criteria: 690804 g/mg
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IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Solution: 15 g/mL in acetonitrile C. PROCEDURE Sample: 1 mg Analysis: Dissolve in 4 mL of water. Add 1 mL of 1 N sulfuric acid while cooling in an ice bath, add 1 mL of a freshly prepared solution of sodium nitrite (1 in 100), allow to stand for 2 min, then add 1 mL of ammonium sulfamate solution (1 in 100). Allow to stand for 1 min, and add 1 mL of N-(1-naphthyl)ethylenediamine dihydrochloride TS. Acceptance criteria: A red-purple color develops. ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.02 M ammonium acetate (2:3) Diluent: Acetonitrile and water (2:3) Standard stock solution: 5 mg/mL of USP Cefpodoxime Proxetil RS in methanol Standard solution: 0.025 mg/mL from the Standard stock solution diluted with Diluent [NOTEPass through a filter having a 0.45-m or finer porosity.] Sample stock solution: 5 mg/mL of Cefpodoxime Proxetil in methanol Sample solution: 0.025 mg/mL from the Sample stock solution diluted with Diluent [NOTEPass through a filter having a 0.45-m or finer porosity.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 30 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTERelative retention times are about 0.9 for cefpodoxime proxetil S-epimer and 1.0 for cefpodoxime proxetil R-epimer.] Suitability requirements Resolution: NLT 2.5 between cefpodoxime proxetil Sepimer and cefpodoxime proxetil R-epimer Tailing factor: NMT 1.5 for cefpodoxime proxetil Repimer Relative standard deviation: NMT 1.0% from the sum of the areas of the cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-epimer peaks Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C15H17N5O6S2 in each mg of Cefpodoxime Proxetil: Result = (rU/rT) (CS/CU) F rU rT CS CU F = sum of the peak responses for cefpodoxime proxetil S-epimer and cefpodoxime proxetil Repimer of the Sample solution = sum of the peak responses for cefpodoxime proxetil S-epimer and cefpodoxime proxetil Repimer of the Standard solution = concentration of USP Cefpodoxime Proxetil RS in the Standard solution (g/mL) = concentration of Cefpodoxime Proxetil in the Sample solution (g/mL) = correction factor, 1000 g/mg
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Solution A: 0.02 M ammonium acetate Solution B: Acetonitrile Mobile Phase: See the gradient table below.
Time (min) 0 10 40 80 85 90 95 100 Solution A (%) 90 68 68 50 50 25 25 90 Solution B (%) 10 32 32 50 50 75 75 10
Diluent: Acetonitrile and water (1:2) System suitability solution: 10 g/mL of USP Cefpodoxime Proxetil RS in Diluent [NOTEA volume of methanol not exceeding 10% of the total volume in the final solution may be used to facilitate dissolution.] Sample stock solution: 10 mg/mL of Cefpodoxime Proxetil in methanol [NOTESonicate if necessary.] Sample solution: 1.0 mg/mL of Cefpodoxime Proxetil from the Sample stock solution diluted with Diluent [NOTEThis solution should be injected promptly, but may be analyzed within 24 h when stored at 8.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 260 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 30 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times are about 0.9 for cefpodoxime proxetil S-epimer and 1.0 for cefpodoxime proxetil R-epimer. The retention time for the cefpodoxime proxetil R-epimer is between 37 and 42 min.] Suitability requirements Resolution: NLT 4.0 between cefpodoxime proxetil Sepimer and cefpodoxime proxetil R-epimer Column efficiency: NLT 19,000 theoretical plates determined from the cefpodoxime proxetil R-epimer peak Relative standard deviation: NMT 2.0% from the sum of the areas of the cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-epimer peaks Analysis Samples: Sample solution Calculate the percentage of each impurity in the portion of Cefpodoxime Proxetil taken: Result = (rU/rT) 100
USP 32
rU = peak area for each impurity = sum of the areas of all the peaks rT Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 6.0%, impurity peaks of less than 0.05% being disregarded
Impurity Table 1 Relative Retention Time 0.86 1.271.39 >2.0 Acceptance Criteria, NMT(%) 3.0 1.0 1.0 0.5
Official Monographs / Cefpodoxime 161

labeling. Shake the resulting suspension thoroughly, and determine its density. Transfer a weighed quantity of the suspension, nominally equivalent to 50 mg of cefpodoxime, to a 100-mL volumetric flask. Add 10 mL of water, and shake to disperse. Add 20 mL of acetonitrile, and sonicate for 15 min. Cool to room temperature, and dilute with Diluent to volume. Sample solution: 0.025 mg/mL from the Sample stock solution diluted with Diluent [NOTEPass through a filter having a 0.45-m or finer porosity.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 30 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-epimer are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between cefpodoxime proxetil Sepimer and cefpodoxime proxetil R-epimer Tailing factor: NMT 1.5 for cefpodoxime proxetil Repimer Relative standard deviation: NMT 1.0% from the sum of the areas of the cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-epimer peaks for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H17N5O6S2 in the portion of Oral Suspension taken: Result = (rU/rS) (CS/CU) P F 100 = sum of the peak responses for cefpodoxime proxetil S-epimer and cefpodoxime proxetil Repimer from the Sample solution = sum of the peak responses for cefpodoxime rS proxetil S-epimer and cefpodoxime proxetil Repimer from the Standard solution = concentration of USP Cefpodoxime Proxetil RS in CS the Standard solution (mg/mL) = nominal concentration of cefpodoxime proxetil CU in the Sample solution (mg/mL) P = stated content of cefpodoxime in USP Cefpodoxime Proxetil RS (g/mg) F = conversion factor, 0.001 mg/g Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905: It meets the requirements for solids packaged in single-unit containers. SPECIFIC TESTS PH 791: 4.05.5, in the suspension constituted as directed in the labeling WATER DETERMINATION 921: NMT 1.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at a temperature not exceeding 30. Store the constituted Oral Suspension in a refrigerator. USP REFERENCE STANDARDS 11 USP Cefpodoxime Proxetil RS rU
Impurity Name Individual impurities Individual impurities Individual impurities All other individual impurities
SPECIFIC TESTS SPECIFIC ROTATION 781S: +35.0 to +48.0 Sample solution: 10 mg/mL, in methanol WATER DETERMINATION, Method I 921: NMT 3.0% ISOMER RATIO Analysis: Using the chromatogram of the Sample solution obtained in the Assay, calculate the ratio of the cefpodoxime proxetil R-epimer peak response to the sum of the peak responses of the cefpodoxime proxetil S-epimer peak and the cefpodoxime proxetil R-epimer peak. Acceptance criteria: The ratio is between 0.5 and 0.6. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at a temperature not exceeding 25. USP REFERENCE STANDARDS 11 USP Cefpodoxime Proxetil RS
Cefpodoxime Proxetil for Oral Suspension

(Comment on this Monograph)id=m14112=Cefpodoxime Proxetil for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cefpodoxime Proxetil for Oral Suspension contains Cefpodoxime Proxetil and one or more buffers, suspending agents, sweeteners, flavorings, and preservatives. When constituted as directed in the labeling, it contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of cefpodoxime (C15H17N5O6S2). IDENTIFICATION The retention times of the cefpodoxime proxetil R-epimer peak and the cefpodoxime proxetil S-epimer peak of the Sample solution correspond to those in the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.02 M ammonium acetate (2:3) Diluent: Acetonitrile and water (2:3) Standard stock solution: 6 mg/mL USP Cefpodoxime Proxetil RS in methanol Standard solution: 0.03 mg/mL USP Cefpodoxime Proxetil RS from Standard stock solution diluted with Diluent [NOTEPass through a filter having a 0.45-m or finer porosity.] Sample stock solution: Constitute a container of Cefpodoxime Proxetil for Oral Suspension as directed in the
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Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H17N5O6S2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) P F 100 = sum of the peak responses for cefpodoxime proxetil S-epimer and cefpodoxime proxetil Repimer from the Sample solution rS = sum of the peak responses for cefpodoxime proxetil S-epimer and cefpodoxime proxetil Repimer from the Standard solution = concentration of USP Cefpodoxime Proxetil RS in CS the Standard solution (mg/mL) = nominal concentration of cefpodoxime proxetil CU in the Sample solution (mg/mL) P = potency of cefpodoxime in USP Cefpodoxime Proxetil RS (g/mg) F = conversion factor, 0.001 mg/g Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION 711 Solution A: 54.5 g of glycine and 42.6 g of sodium chloride in about 500 mL of water in a 1000-mL volumetric flask Cautiously add, with swirling, 14.2 mL of hydrochloric acid, and allow to cool. Dilute with water to volume, and mix. Transfer 50 mL of this stock solution to a flask, and dilute with water to 900 mL to obtain a solution having a pH of 3.0 0.1. [NOTEIf necessary, adjust the pH of the stock solution with 10 N sodium hydroxide so that when 50 mL is diluted with water to 900 mL, the pH of the Dissolution Medium is 3.0 0.1.] Medium: Solution A, 900 mL Apparatus 2: 75 rpm Time: 30 min Sample solutions: Sample per Dissolution 711, filtered Standard solution: USP Cefpodoxime Proxetil RS dissolved in a small volume of methanol, and diluted with Medium to a known concentration Analysis Sample: Standard solution and Sample solution Determine the amount of C15H17N5O6S2 dissolved by employing UV absorption at 259 nm on the Sample solution in comparison with the Standard solution. Tolerances: NLT 70% (Q) of the labeled amount of C15H17N5O6S2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION 921: NMT 5.0%
Cefpodoxime Proxetil Tablets

(Comment on this Monograph)id=m14113=Cefpodoxime Proxetil Tablets=Ca-Chl-Monos.pdf) DEFINITION Cefpodoxime Proxetil Tablets contain an amount of Cefpodoxime Proxetil equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of cefpodoxime (C15H17N5O6S2). IDENTIFICATION The retention times of the cefpodoxime proxetil R-epimer peak and the cefpodoxime proxetil S-epimer peak of the Sample solution correspond to those in the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.02 M ammonium acetate (2:3) Diluent: Acetonitrile and water (2:3) Standard stock solution: Transfer about 30mg of USP Cefpodoxime Proxetil RS to a 50-mL volumetric flask, dissolve in 5 mL of methanol, dilute with Diluent to volume, and mix. Standard solution: 0.03 mg/mL USP Cefpodoxime Proxetil RS from the Standard stock solution diluted with Diluent [NOTEPass through a filter having a 0.45-m or finer porosity.] Sample stock solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder, nominally equivalent to 50 mg of cefpodoxime, to a 100-mL volumetric flask. Dissolve in 40 mL of Diluent, sonicating for 5 min. Cool to room temperature, and dilute with Diluent to volume. Sample solution: 0.025 mg/mL from the Sample stock solution, diluted with Diluent [NOTEPass through a filter having a 0.45-m or finer porosity.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 235 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 30 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for cefpodoxime proxetil Sepimer and cefpodoxime proxetil R-epimer are 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between cefpodoxime proxetil Sepimer and cefpodoxime proxetil R-epimer Tailing factor: NMT 1.5 for cefpodoxime proxetil Repimer Relative standard deviation: NMT 1.0% from the sum of the areas of the cefpodoxime proxetil S-epimer and cefpodoxime proxetil R-epimer peaks for replicate injections
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cefpodoxime Proxetil RS
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Official Monographs / Cefprozil 163

Suitability requirements Resolution: NLT 2.5 between the cefprozil (Z)-isomer peak and the cefprozil (E)-isomer peak, System suitability solution Column efficiency: NLT 2500 theoretical plates in Standard solution A from the cefprozil (Z)-isomer peak, when calculated: Result = 5.545(tr/Wh/2)2
Cefprozil
(Comment on this Monograph)id=m14115=Cefprozil=Ca-ChlMonos.pdf)
C18H19N3O5S H2O 407.44 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[amino(4-hydroxyphenyl)acetyl]amino]-8-oxo-3-(1-propenyl)monohydrate, [6R-[6, 7(R*)]]-; (6R,7R)-7-[(R)-2-Amino-2-(p-hydroxyphenyl)acetamido]-8-oxo-3propenyl-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid [121123-17-9]; Anhydrous 389.43 [92665-29-7]. DEFINITION Cefprozil contains NLT 900 g and NMT 1050 g of cefprozil (C18H19N3O5S)/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Standard solution: USP Cefprozil (Z)-Isomer RS B. The retention times of the cefprozil (Z)-isomer and cefprozil (E)-isomer peaks of the Sample solution correspond to those of the Standard solutions, as obtained in the Assay. ASSAY PROCEDURE Solution A: 11.5 mg/mL of monobasic ammonium phosphate in water Adjust, if necessary, with phosphoric acid to a pH of 4.4. Mobile phase: Acetonitrile and Solution A (1:9) Pass this solution through a filter having a porosity of 0.5 m or finer. [NOTEDecreasing the proportion of acetonitrile increases retention times and improves the separation of the cefprozil isomer peaks.] Standard solution A: 0.25 mg/mL of USP Cefprozil (Z)Isomer RS [NOTEUse this solution within 6 h.] Standard stock solution B: 0.25 mg/mL of USP Cefprozil (E)-Isomer RS [NOTEUse this solution within 6 h.] Standard solution B: 0.025 mg/mL from Standard stock solution B diluted with water System suitability solution: Standard solution A and Standard stock solution B (1:1) [NOTEUse this solution within 6 h.] Sample solution: 0.3 mg/mL of Cefprozil in water Shake to assure dissolution. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution A [NOTEThe relative retention times for cefprozil (Z)-isomer and cefprozil (E)-isomer are about 0.7 and 1.0, respectively.]
Tailing factor: 0.91.1, in Standard solution A from the cefprozil (Z)-isomer peak, when calculated: Result = W0.1/2f W0.1 = width of the peak at 10% height Relative standard deviation: NMT 2.0% in Standard solution A Analysis Samples: Standard solution A, Standard solution B, and Sample solution B, Calculate the quantity (g) of cefprozil (Z)-isomer and cefprozil (E)-isomer in each mg of Cefprozil taken: Result = (rU/rS) (CS/CU) P = peak response of the cefprozil (Z)-isomer or the cefprozil (E)-isomer, as appropriate, of the Sample solution rS = peak response of the cefprozil (Z)-isomer or the cefprozil (E)-isomer, as appropriate, of the Standard solution CS = concentration of USP Cefprozil (Z)-isomer RS in Standard solution A or of the USP Cefprozil (E)isomer RS in Standard solution B, as appropriate (mg/mL) = nominal concentration of Cefprozil taken to CU prepare the Sample solution (mg/mL) P = the assigned potency (g/mg) of the appropriate USP Reference Standard Calculate the quantity, in g, of C18H19N3O5S in each mg of the Cefprozil taken by adding the values, in g/mg, of the cefprozil (Z)-isomer and the cefprozil (E)-isomer. Acceptance criteria: 9001050 g/mg rU SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 3.56.5, in a solution containing 5 mg/mL WATER DETERMINATION, Method I 921: 3.5%6.5% CEFPROZIL (E)-ISOMER RATIO Analysis: Calculate the ratio of the cefprozil (E)-isomer to the total cefprozil taken: Result = E/(E + Z) = content of cefprozil (E)-isomer (g/mg) as determined in the Assay Z = content of cefprozil (Z)-isome (g/mg) as determined in the Assay Acceptance criteria: The ratio is 0.060.11. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefprozil (E)-Isomer RS USP Cefprozil (Z)-Isomer RS E
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Sample solution: Transfer 15.0 mL of the Sample stock solution to a 50-mL volumetric flask, dilute with water to volume, and mix. Pass a portion of this solution through a filter having a 0.5-m or finer porosity. [NOTEUse this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution A [NOTERelative retention times are about 0.7 for the cefprozil (Z)-isomer and 1.0 for the cefprozil (E)-isomer.] Suitability requirements Resolution: NLT 2.5, between the cefprozil (Z)-isomer peak and the cefprozil (E)-isomer peak, System suitability solution Column efficiency: NLT 2500 theoretical plates from the cefprozil (Z)-isomer peak, Standard solution A, when calculated: Result = 5.545(tr/Wh/2)2 Tailing factor: 0.91.1 from the cefprozil (Z)-isomer peak, Standard solution A when calculated: Result = W0.1/2f = width of the peak at 10% height W0.1 Relative standard deviation: NMT 2.0%, Standard solution A Analysis Samples: Sample solution and Standard solution Calculate the concentration of the cefprozil (Z)-isomer and the cefprozil (E)-isomer (mg/mL) in the Sample solution taken: Result = (rU/rS) CS P F = peak response of the cefprozil (Z)-isomer or the cefprozil (E)-isomer, as appropriate, of the Sample solution rS = peak response of the cefprozil (Z)-isomer or the cefprozil (E)-isomer, as appropriate, of the Standard solution CS = concentration of USP Cefprozil (Z)-Isomer RS in Standard solution A or of the USP Cefprozil (E)Isomer RS in Standard solution B, as appropriate (mg/mL) P = the assigned potency of the appropriate USP Reference Standard (g/mg) F = correction factor, 0.001 mg/g Calculate the percentage of C18H19N3O5S in the portion of Cefprozil for Oral Suspension taken: rU Result = 100 (CZ + CE)/CU = concentration of the cefprozil (Z)-isomer in the Sample solution (mg/mL) CE = concentration of the cefprozil (E)-isomer in the Sample solution (mg/mL) = nominal concentration of cefprozil in the Sample CU solution (mg/mL) Acceptance criteria: 90%120.0% CZ PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: It meets the requirements for solids packaged in single-unit containers.
Cefprozil for Oral Suspension

(Comment on this Monograph)id=m14117=Cefprozil for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cefprozil for Oral Suspension is a dry mixture of Cefprozil and one or more suitable buffers, flavors, preservatives, suspending agents, and sweeteners. It contains NLT 90.0% and NMT 120.0% of the labeled amount of cefprozil (C18H19N3O5S). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 5 mg/mL of USP Cefprozil (Z)-Isomer RS in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Sample solution: Equivalent to 50 mg of cefprozil from Cefprozil for Oral Suspension powder, in a 20-mL glassstoppered test tube Add 10 mL of a mixture of acetone and 0.1 N hydrochloric acid (4:1), shake for 5 min, and allow to settle. Use the supernatant. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Butyl alcohol, glacial acetic acid, and water (60:20:20) Analysis Samples: Sample solution and Standard solution Allow the spots to dry, and develop the chromatogram in an equilibrated chromatographic chamber with the solvent system, until the solvent front has moved threefourths of the length of the plate. Remove the plate from the chamber, and allow the plate to air-dry in a hood. Place the dry plate in a chamber containing iodine vapors. Examine the plate, and locate the spots. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. The retention times of the cefprozil (Z)-isomer and cefprozil (E)-isomer peaks of the Sample solution correspond to those of the Standard solutions, as obtained in the Assay. ASSAY PROCEDURE Solution A: 11.5 mg/mL of monobasic ammonium phosphate in water Adjust, if necessary, with phosphoric acid to a pH of 4.4. Mobile phase: Acetonitrile and Solution A (1:9) Pass this solution through a filter having a porosity of 0.5 m or finer. [NOTEDecreasing the proportion of acetonitrile increases retention times and improves the separation of the cefprozil isomer peaks.] Standard solution A: 0.25 mg/mL of USP Cefprozil (Z)Isomer RS [NOTEUse this solution within 6 h.] Standard stock solution B: 0.25 mg/mL of USP Cefprozil (E)-Isomer RS Standard solution B: 0.025 mg/mL from Standard stock solution B diluted with water [NOTEUse this solution within 6 h.] System suitability solution: Standard solution A and Standard stock solution B (1:1) [NOTEUse this solution within 6 h.] Sample stock solution: Constitute one container of Cefprozil for Oral Suspension as directed in the labeling. Transfer a volume of Cefprozil for Oral Suspension, freshly mixed and free from air bubbles, nominally equivalent to 250 mg of cefprozil, to a 250-mL volumetric flask, dilute with water to volume, and mix, sonicating briefly.
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DELIVERABLE VOLUME 698: It meets the requirements for powder packaged in single-unit containters. SPECIFIC TESTS PH 791: 4.06.0, in the Cefprozil for Oral Suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 3.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefprozil (E)-Isomer RS USP Cefprozil (Z)-Isomer RS
Official Monographs / Cefprozil 165

Standard solution B: 0.025 mg/mL from Standard stock solution B, diluted with water [NOTEUse this solution within 6 h.] System suitability solution: Standard solution A and Standard stock solution B (1:1) [NOTEUse this solution within 6 h.] Sample stock solution: Transfer a number of Tablets, nominally equivalent to 1500 mg of cefprozil, to a 250-mL volumetric flask containing 180 mL of water. Allow the Tablets to disintegrate with the aid of swirling and sonication. Dilute with water to volume. Sample solution: Nominally 0.3 mg/mL of cefprozil from the Sample stock solution diluted with water [NOTEPass a portion of this solution through a filter having a 0.5-m or finer porosity, and use the filtrate as the Sample solution. Use this solution within 6 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution A [NOTEThe relative retention times for cefprozil (Z)-isomer and cefprozil (E)-isomer are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.5 between the cefprozil (Z)-isomer peak and the cefprozil (E)-isomer peak, System suitability solution Column efficiency: NLT 2500 theoretical plates in Standard solution A from the cefprozil (Z)-isomer peak, when calculated: Result = 5.545(tr/Wh/2)2 Tailing factor: 0.91.1 in Standard solution A from the cefprozil (Z)-isomer peak, when calculated: Result = W0.1/2f = width of the peak at 10% height W0.1 Relative standard deviation: NMT 2.0% in Standard solution A Analysis Samples: Sample solution and Standard solution Calculate the concentration of the cefprozil (Z)-isomer and cefprozil (E)-isomer, in mg/mL, of the Sample solution taken: Result = (rU/rS) CS P F = peak response of the cefprozil (Z)-isomer or the cefprozil (E)-isomer, as appropriate, from the Sample solution rS = peak response of the cefprozil (Z)-isomer or the cefprozil (E)-isomer, as appropriate, from the Standard solution = concentration of USP Cefprozil (Z)-Isomer RS in CS Standard solution A or of the USP Cefprozil (E)Isomer RS in Standard solution B, as appropriate (mg/mL) P = assigned potency of the appropriate USP Reference Standard (g/mg) F = correction factor, 0.001 mg/g Calculate the percentage of label claim of C18H19N3O5S in the portion of Tablets taken: rU Result = (CZ + CE)/CU 100
Cefprozil Tablets
(Comment on this Monograph)id=m14118=Cefprozil Tablets=Ca-Chl-Monos.pdf) DEFINITION Cefprozil Tablets contain NLT 90.0% and NMT 120.0% of the labeled amount of cefprozil (C18H19N3O5S). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 5 mg/mL of USP Cefprozil (Z)-Isomer RS in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Sample solution: Equivalent to 2.5 mg/mL of cefprozil from powdered Tablets in a mixture of acetone and 0.1 N hydrochloric acid (4:1) Shake for 5 min, and allow the mixture to settle. Use the supernatant as the Sample solution. Chromatographic system (See Chromatography 621, Thin-layer Chromatography.) Mode: TLC Application volume: 10 L Developing solvent system: Butyl alcohol, glacial acetic acid, and water (60:20:20) Analysis Samples: Sample solution and Standard solution Allow the spots to dry, and develop the chromatogram in an equilibrated chromatographic chamber with the solvent system, until the solvent front has moved threefourths of the length of the plate. Remove the plate from the chamber, and allow the plate to air-dry in a hood. Place the dry plate in a chamber containing iodine vapors. Examine the plate, and locate the spots. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. B. The retention times of the cefprozil (Z)-isomer and cefprozil (E)-isomer peaks of the Sample solution correspond to those of the Standard solutions, as obtained in the Assay. ASSAY PROCEDURE Solution A: 11.5 mg/mL of monobasic ammonium phosphate in water Adjust, if necessary, with phosphoric acid to a pH of 4.4. Mobile phase: Acetonitrile and Solution A (1:9) Filter this solution through a filter having a porosity of 0.5 m or finer. [NOTEDecreasing the proportion of acetonitrile increases retention times and improves the separation of the cefprozil isomer peaks.] Standard solution A: 0.25 mg/mL of USP Cefprozil (Z)Isomer RS [NOTEUse this solution within 6 h.] Standard stock solution B: 0.25 mg/mL of USP Cefprozil (E)-Isomer RS
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CZ
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= concentration of the cefprozil (Z)-isomer in the Sample solution (mg/mL) CE = concentration of the cefprozil (E)-isomer in the Sample solution (mg/mL) CU = nominal concentration of the cefprozil in the Sample solution (mg/mL) Acceptance criteria: 90%120.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 45 min Analysis: Determine the amount of C18H19N3O5S dissolved in the Dissolution Medium, as directed in the Assay, using instead of the Assay Sample solution, a filtered portion of the Dissolution Medium, diluted if necessary, to obtain a Sample solution containing 0.3 mg of cefprozil/mL. Calculate the quantity, in mg, of cefprozil (Z)-isomer and cefprozil (E)-isomer dissolved: Result = V (rU/rS) CS P F D = volume of Medium in mL, 900 = peak response of the cefprozil (Z)-isomer or the cefprozil (E)-isomer, as appropriate, from the Sample solution = peak response of the cefprozil (Z)-isomer or the rS cefprozil (E)-isomer, as appropriate, from the Standard solution = concentration of USP Cefprozil (Z)-Isomer RS in CS Standard solution A or of USP Cefprozil (E)Isomer RS in Standard solution B, as appropriate (mg/mL) F = correction factor, 0.001 mg/g P = assigned potency of the appropriate USP Reference Standard (g/mg) D = 1 or, where the filtered Dissolution Medium was diluted to prepare the Sample solution, the appropriate dilution factor Calculate the percentage of label claim of C18H19N3O5S dissolved: Result = (MZ + ME) 100/L = quantity of cefprozil (Z)-isomer dissolved (mg) MZ = quantity of cefprozil (E)-isomer dissolved (mg) ME L = label claim of cefprozil in Tablet (mg) Tolerances: NLT 75% (Q) of the labeled amount of cefprozil (C18H19N3O5S) UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 7.0% V rU
Ceftazidime
(Comment on this Monograph)id=m14120=Ceftazidime=CaChl-Monos.pdf)
C22H22N6O7S2 5H2O 636.65 Pyridinium, 1-[[7-[[(2-amino-4-thiazolyl)[(1-carboxy-1methylethoxy)imino]acetyl]amino]-2-carboxy-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-, hydroxide, inner salt, pentahydrate, [6R[6,7 (Z)]]-; 1-[[(6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-2carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en3yl]methyl]pyridinium hydroxide, inner salt, 72-(Z)-[O-(1carboxy-1-methylethyl)oxime], pentahydrate [78439-06-2]. Anhydrous 546.59 DEFINITION Ceftazidime contains NLT 95.0% and NMT 102.0% of C22H22N6O7S2, calculated on the dried basis. IDENTIFICATION The retention time of the Standard solution corresponds to that of the major peak for ceftazidime from the Sample solution as obtained in the Assay. ASSAY PROCEDURE Buffer solution: 42.59 mg/mL of anhydrous dibasic sodium phosphate and 27.22 mg/mL of monobasic potassium phosphate in water Mobile phase: Mix 40 mL of acetonitrile and 200 mL of Buffer solution, and dilute with water to obtain 2000 mL of solution. Filter using a filter having a porosity of 1 m or finer, and degas. Standard stock solution: Transfer 29 mg of USP Ceftazidime Pentahydrate RS to a 25-mL volumetric flask containing 2.5 mL of Buffer solution, and shake until dissolved. Dilute with water to volume. [NOTEProtect this solution from light.] Standard solution: 100 g/mL from the Standard stock solution diluted with water. [NOTEPrepare the solution immediately prior to chromatography.] System suitability stock solution: 0.1 mg/mL of USP Ceftazidime, Delta-3-Isomer RS in Buffer solution System suitability solution: Mix 1 mL of System suitability stock solution with 8 mL of water and 1 mL of the Standard stock solution. [NOTEPrepare immediately prior to chromatography.] Sample stock solution: Transfer 115 mg of Ceftazidime to a 100-mL volumetric flask containing 10.0 mL of Buffer
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cefprozil (E)-Isomer RS USP Cefprozil (Z)-Isomer RS
USP 32
solution, and shake until dissolved. Dilute with water to volume. [NOTEProtect this solution from light.] Sample solution: Dilute 5.0 mL of Sample stock solution to 50 mL with water. [NOTEPrepare immediately prior to chromatography.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 2.0 between ceftazidime and ceftazidime, delta-3-isomer, System suitability solution Tailing factor: 0.751.5, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H22N6O7S2 in the portion taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of Ceftazidime in the Standard solution (g/mL) CU = concentration of Ceftazidime in the Sample solution (g/mL) Acceptance criteria: 95.0%102.0% rU rS CS SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements STERILITY TESTS 71: Where the label states that it is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration, except to use Fluid A. [NOTETo each 1000 mL of Fluid A, 10 g of sodium bicarbonate have been added before sterilization.] PH 791: 3.04.0, in a solution containing 5 mg/mL LOSS ON DRYING 731 Sample: 300 mg Analysis: Dry in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h. Acceptance criteria: It loses 13.0%15.0% of its weight. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Ceftazidime is sterile or that it must be subjected to further processing during the preparation of injectable or other sterile dosage forms, it contains NMT 0.1 USP Endotoxin Unit/mg of ceftazidime. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable or other sterile dosage forms. USP REFERENCE STANDARDS 11 USP Ceftazidime Delta-3-Isomer RS USP Ceftazidime Pentahydrate RS USP Endotoxin RS
Official Monographs / Ceftazidime 167
Ceftazidime Injection
(Comment on this Monograph)id=m14122=Ceftazidime Injection=Ca-Chl-Monos.pdf) DEFINITION Ceftazidime Injection is a sterile isoosmotic solution of Ceftazidime in Water for Injection. It contains one or more suitable buffers and a tonicity-adjusting agent. It contains NLT 90.0% and NMT 120.0% of the labeled amount of C22H22N6O7S2. IDENTIFICATION The Sample solution exhibits a major peak for ceftazidime, the retention time of which corresponds to that exhibited in the Standard solution obtained as directed in the Assay. ASSAY PROCEDURE Buffer solution: 42.59 mg/mL of anhydrous dibasic sodium phosphate and 27.22 mg/mL of monobasic potassium phosphate in water Mobile phase: Mix 40 mL of acetonitrile and 200 mL of Buffer solution, and dilute with water to obtain 2000 mL of solution. Filter, using a filter having a porosity of 1 m or finer, and degas. Standard stock solution: Transfer 29 mg of USP Ceftazidime Pentahydrate RS to a 25-mL volumetric flask containing 2.5 mL of Buffer solution, and shake until dissolved. Dilute with water to volume. [NOTEProtect this solution from light.] Standard solution: 100 g/mL from the Standard stock solution diluted with water [NOTEPrepare the solution immediately before chromatography.] System suitability stock solution: 0.1 mg/mL of USP Ceftazidime, Delta-3-Isomer RS in Buffer solution System suitability solution: Mix 1 mL of System suitability stock solution with 8 mL of water and 1 mL of the Standard stock solution. [NOTEPrepare immediately before chromatography.] Sample stock solution: Allow a container of the Injection to thaw, and mix the solution. Transfer a volume of the Injection, nominally equivalent to 50 mg of ceftazidime, to a 50-mL volumetric flask, and dilute with Buffer solution to volume. Sample solution: Dilute 5.0 mL of Sample stock solution to 50 mL with water. [NOTEPrepare immediately before chromatography.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 2.0 between ceftazidime and ceftazidime, delta-3-isomer, System suitability solution Tailing factor: 0.751.5, Standard solution Relative standard deviation: NMT 1.0%, Standard solution
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Ceftazidime / Official Monographs
USP 32
containing 2.5 mL of Buffer solution, and shake until dissolved. Dilute with water to volume. [NOTEProtect this solution from light.] Standard solution: 100 g/mL from the Standard stock solution diluted with water [NOTEPrepare the solution immediately prior to use.] System suitability stock solution: 0.1 mg/mL of USP Ceftazidime, Delta-3-Isomer RS in the Buffer solution System suitability solution: Mix 1 mL of System suitability stock solution with 8 mL of water and 1 mL of the Standard stock solution. [NOTEPrepare immediately prior to use.] Sample stock solution A: Transfer a quantity of Ceftazidime for Injection, nominally equivalent to 250 mg of ceftazidime, to a 250-mL volumetric flask, and dilute with water to volume. [NOTEProtect this solution from light.] Sample solution A: Dilute 5.0 mL of Sample stock solution A with water to 50 mL. [NOTEPrepare immediately prior to use.] Sample stock solution B (where it is represented as being in a single-dose container): Constitute a container of Ceftazidime for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with water to obtain a solution containing nominally 1 mg of ceftazidime/mL. [NOTEProtect this solution from light.] Sample solution B: Dilute 5.0 mL of Sample stock solution B with water to 50 mL. [NOTEPrepare immediately prior to use.] Sample stock solution C (where the label states the quantity of ceftazidime in a given volume of constituted solution): Constitute a container of Ceftazidime for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a volume of the constituted solution with water to obtain a solution containing nominally 1 mg of ceftazidime/mL. [NOTEProtect this solution from light.] Sample solution C: Dilute 5.0 mL of Sample stock solution C with water to 50 mL. [NOTEPrepare immediately prior to use.] Chromatographic system: (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 2.0 between ceftazidime and ceftazidime, delta-3-isomer, System suitability solution Tailing factor: 0.751.5, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis I. Samples: Standard solution and Sample solution A Calculate the percentage of C22H22N6O7S2 on the dried and sodium carbonate-free or arginine-free basis in the portion of Ceftazidime for Injection taken: Result = (rU/rS) {C/[W (100-m-s)]} F V D 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H22N6O7S2 in the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of ceftazidime in the Standard solution (g/mL) = nominal concentration of ceftazidime in the CU Sample solution (g/mL) Acceptance criteria: 90.0%120.0% rU rS CS SPECIFIC TESTS PYROGEN TEST 151: Meets the requirements, the test dose being a volume of undiluted Injection providing 80 mg of ceftazidime/kg STERILITY TESTS 71: Meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration PH 791: 5.07.5 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements for Injections 1, Labeling. The label states that it is to be thawed just before use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Ceftazidime Delta-3-Isomer RS USP Ceftazidime Pentahydrate RS
Ceftazidime for Injection

(Comment on this Monograph)id=m14124=Ceftazidime for Injection=Ca-Chl-Monos.pdf) DEFINITION Ceftazidime for Injection is a sterile mixture of Sterile Ceftazidime and Sodium Carbonate or Arginine. It contains NLT 90.0% and NMT 105.0% of ceftazidime (C22H22N6O7S2) on the dried and sodium carbonate- or arginine-free basis, and NLT 90.0% and NMT 120.0% of the labeled amount of ceftazidime (C22H22N6O7S2). IDENTIFICATION A. The retention time of the major peak for ceftazidime from the Sample solutions corresponds to that of the Standard solution, as obtained in the Assay. B. It dissolves in 1 N hydrochloric acid with effervescence, evolving a colorless gas, which when passed into calcium hydroxide TS produces a white precipitate immediately. ASSAY PROCEDURE Buffer solution: 42.59 mg/mL of anhydrous dibasic sodium phosphate and 27.22 mg/mL of monobasic potassium phosphate in water Mobile phase: Mix 40 mL of acetonitrile and 200 mL of Buffer solution, and dilute with water to obtain 2000 mL of solution. Filter, using a filter having a porosity of 1 m or finer, and degas. Standard stock solution: Transfer 29 mg of USP Ceftazidime Pentahydrate RS to a 25-mL volumetric flask
USP 32
= concentration of C22H22N6O7S2 in the Standard solution (mg/mL) W = mg of Ceftazidime for Injection taken to prepare Sample stock solution A m = total percentage of loss on drying s = percentage of sodium carbonate or arginine in the Ceftazidime for Injection taken F = correction factor 100 V = volume of Sample stock solution A, 250 mL D = dilution factor to prepare Sample solution A from Sample stock solution A, 10 Acceptance criteria: 90.0%105.0% II. Samples: Sample solution B or Sample solution C, and Standard solution Calculate the percentage of C22H22N6O7S2 withdrawn from the container or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ceftazidime Pentahydrate RS in the Standard solution (g/mL) = nominal concentration of ceftazidime in Sample CU solution B or Sample solution C (g/mL) Acceptance criteria: 90.0%120.0% rU rS CS OTHER COMPONENTS SODIUM CARBONATE (WHERE PRESENT) Solution A: 19.07 mg/mL of potassium chloride Standard stock solution: 14 g/mL of sodium chloride, previously dried at 105 for 2 h Standard solution: Transfer 10.0 mL of Standard stock solution to a 100-mL volumetric flask, add 10.0 mL of Solution A, and dilute with water to volume. Blank: Transfer 10.0 mL of Solution A to a 100-mL volumetric flask, and dilute with water to volume. Sample stock solution: Use Sample stock solution A, and dilute an aliquot with water to obtain a solution containing 12.5 g of sodium carbonate per mL. Sample solution: Transfer 10.0 mL of Sample stock solution to a 100-mL volumetric flask, add 10.0 mL of Solution A and dilute with water to volume. Spectrophotometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer equipped with a sodium hollow-cathode lamp and an airacetylene flame Analytical wavelength: 589.0 nm (the sodium emission line) Analysis Samples: Standard solution and Sample solution Calculate the percentage of Na2CO3 in the portion of Ceftazidime for Injection taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 AU AS CS = absorbance of the Sample solution = absorbance of the Standard solution = concentration of sodium chloride in the Standard solution (g/mL) = concentration of Ceftazidime for Injection in the CU Sample solution (g/mL) = molecular weight of sodium carbonate, 105.99 Mr1 = twice the molecular weight of sodium chloride, Mr2 116.88 [NOTEUse this percentage to calculate the result from Sample solution A.] CS
Official Monographs / Ceftazidime 169

CONTENT OF ARGININE (WHERE PRESENT) Solution A: 1.15 mg/mL of monobasic ammonium phosphate in water, adjusted with phosphoric acid to a pH of 2.0 0.1 Mobile phase: Acetonitrile and Solution A (3:1) Standard solution: 0.2 mg/mL each of USP Ceftazidime Pentahydrate RS and of USP L-Arginine RS Sample solution: 0.2 mg/mL from Ceftazidime for Injection diluted with water Chromatographic system: (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 206 nm Column: 4-mm 25-cm; packing L20 Saturator pre-column: 4.6-mm 50-cm; packing L27 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 6.0 between the ceftazidime and the arginine peaks Tailing factor: NMT 4.0 for the arginine peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H14N4O2 in the Ceftazidime for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of arginine from the Sample solution = peak response of arginine from the Standard rS solution = concentration of USP L-Arginine RS in the CS Standard solution (mg/mL) = concentration of Ceftazidime for Injection in the CU Sample solution (mg/mL) [NOTEUse this percentage to calculate the assay result from Sample solution A.] PERFORMANCE TESTS Uniformity of Dosage Units 905: requirements Meets the rU
IMPURITIES Organic Impurities LIMIT OF PYRIDINE Solution A: 5.68 mg/mL of anhydrous dibasic sodium phosphate and 3.63 mg/mL of monobasic potassium phosphate in water Mobile phase: Acetonitrile, water, and 0.25 M monobasic ammonium phosphate (3:6:1), adjusted with ammonium hydroxide to a pH of 7.0 0.1 Pass this solution through a filter having a 1-m or finer porosity. (See Chromatography 621, System Suitability.) Standard stock solution: 2.5 mg/mL of pyridine in water Standard solution: 25 g/mL from the Standard stock solution in Solution A. [NOTEPrepare immediately prior to use.] Sample solution: To 660 mg of Ceftazidime for Injection, just removed from its container, in a 100-mL volumetric flask, promptly add Solution A to volume, and mix. Store this solution in a cool place, and use it within 1 h. Chromatographic system (See Chromatography 621, System Suitability.)
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Ceftazidime / Official Monographs

USP Ceftazidime Pentahydrate RS USP Endotoxin RS
USP 32
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.6 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.5 in the Standard solution Relative standard deviation: NMT 3.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of pyridine in the portion of Ceftazidime for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of pyridine from the Sample solution RS = peak response of pyridine from the Standard solution CS = concentration of pyridine in the Standard solution (g/mL) CU = concentration of the Sample solution (g/mL) Impurity Acceptance criteria: NMT 0.4% of pyridine is found where it contains sodium carbonate; and NMT 0.3% where it contains arginine. RU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.1 USP Endotoxin Unit/mg of ceftazidime STERILITY TESTS 71: Meets the requirements for Test for Sterility of the Product to be Examined, Membrane Filtration PH 791: 5.07.5, in a solution constituted in the sealed container (taking care to relieve the pressure inside the container during constitution) containing 100 mg of ceftazidime/mL LOSS ON DRYING 731: Dry 300 mg in vacuum at a pressure not exceeding 5 mm of mercury at 25 for 4 h: where it contains arginine, it loses NMT 12.5% of its weight. Where it contains sodium carbonate, it loses NMT 13.5% of its weight. Where it contains arginine, use the percentage loss obtained, m, to calculate the result from Sample solution A. Where it contains sodium carbonate, heat the residue in vacuum at a pressure not exceeding 5 mm of mercury at 100 an additional 3 h, and calculate the total percentage of weight loss. Use this percentage, m, to calculate the result from Sample solution A. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: Meets the requirements for Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids, protected from light. USP REFERENCE STANDARDS 11 USP L-Arginine RS USP Ceftazidime Delta-3-Isomer RS
Ceftizoxime Sodium
(Comment on this Monograph)id=m14139=Ceftizoxime Sodium=Ca-Chl-Monos.pdf)
405.39 C13H12N5NaO5S2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2,3dihydro-2-imino-4-thiazoly)(methoxyimino)acetyl]amino]-8oxomonosodium salt, [6R-[6,7(Z)]]-; Sodium (6R,7R)-7-[2-(2-imino-4-thiazolin-4-yl)glyoxylamido]-8oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylate 72-(Z)(O-methyloxime) [68401-82-1]. DEFINITION Ceftizoxime Sodium contains the equivalent of NLT 850 g and NMT 995 g of ceftizoxime (C13H13N5O5S2)/mg, calculated on the anhydrous basis. IDENTIFICATION A. The chromatogram of the Sample solution exhibits a major peak for ceftizoxime, the retention time of which corresponds to that exhibited by the Standard solution, obtained as directed in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191: Meets the requirements of the tests ASSAY PROCEDURE Buffer A: 1.42 mg/mL of citric acid monohydrate and 1.73 mg/mL of dibasic sodium phosphate in water Buffer B: 3.63 mg/mL of monobasic potassium phosphate and 10.73 mg/mL of dibasic sodium phosphate in water Mobile phase: Acetonitrile and Buffer A (1:9). Filter through a filter of 1 m or finer porosity. [NOTEAdjust the composition, if necessary, to meet the performance requirements under Chromatographic system.] Internal standard solution: Dissolve 1.2 g of salicylic acid in 10 mL of methanol, and dilute with Buffer B to 200 mL. Standard stock solution: 1 mg/mL of USP Ceftizoxime RS in Buffer B Standard solution: Add 2.0 mL of Standard stock solution and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Sample stock solution: 1 mg/mL of Ceftizoxime Sodium in Buffer B Sample solution: Add 2.0 mL of Sample stock solution and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; 5- to 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for ceftizoxime and salicylic acid are about 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 4 between the analyte and internal standard peaks Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Sample: Standard solution and Sample solution Calculate the quantity, in g, of ceftizoxime/mg of the Ceftizoxime Sodium: Result = (RU/RS) (CS/CU) F = peak response ratio of ceftizoxime to the internal standard peak from the Sample solution = peak response ratio of ceftizoxime to the internal RS standard peak from the Standard solution = concentration of USP Ceftizoxime RS in the CS Standard solution (g/mL) = concentration of Ceftizoxime Sodium in the CU Sample solution (g/mL) F = conversion factor, 1000 g/mg Acceptance criteria: 850995 g/mg SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 6.08.0, in a solution (1 in 10) WATER DETERMINATION, Method I 921: NMT 8.5% STERILITY TESTS 71: Where the label states that Ceftizoxime Sodium is sterile, it meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Ceftizoxime Sodium is sterile, or it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.10 USP Endotoxin Unit/mg of ceftizoxime. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Ceftizoxime RS USP Endotoxin RS RU
Official Monographs / Ceftizoxime 171

IDENTIFICATION The retention time of the major peak for ceftizoxime from the Sample solution corresponds to that of the Standard solution as obtained in the Assay. ASSAY PROCEDURE Buffer A: 1.42 mg/mL of citric acid monohydrate and 1.73 mg/mL of dibasic sodium phosphate Buffer B: 3.63 mg/mL of monobasic potassium phosphate and 10.73 mg/mL of dibasic sodium phosphate Mobile phase: Acetonitrile and Buffer A (1:9) Pass through a filter of 1-m or finer porosity. [NOTEAdjust the composition, if necessary, to meet the performance requirements under Chromatographic system.] Internal standard solution: Dissolve 1.2 g of salicylic acid in 10 mL of methanol, and dilute with Buffer B to 200 mL. Standard stock solution: 1 mg/mL of USP Ceftizoxime RS in Buffer B Standard solution: Add 2.0 mL of Standard stock solution and 5.0 mL of Internal standard solution. Dilute with Buffer B to 100 mL. Sample stock solution: Allow 1 container of Injection to thaw, and mix. Transfer a volume of the Injection, nominally equivalent to about 40 mg of ceftizoxime, to a 100-mL volumetric flask, and dilute with Buffer B to volume. Sample solution: To 5.0 mL of Sample stock solution, add 5.0 mL Internal standard solution. Dilute with Buffer B to 100 mL. Chromatographic system: (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; 5- to 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times are about 0.6 for ceftizoxime and 1.0 for salicylic acid.] Suitability requirements Resolution: NLT 4 between the analyte and internal standard peaks. Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Sample solution and Standard solution Calculate the percentage of label claim of C13H13N5O5S2 in the Injection Result = (RU/RS) (CS/CU) 100 = peak response ratio of the ceftizoxime peak to the internal standard peak from the Sample solution = peak response ratio of the ceftizoxime peak to RS the internal standard peak from the Standard solution CS = concentration of USP Ceftizoxime RS in the Standard solution (g/mL) = nominal concentration of ceftizoxime sodium in CU the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: Unit/mg of ceftizoxime NMT 0.10 USP Endotoxin RU
Ceftizoxime Injection
(Comment on this Monograph)id=m14141=Ceftizoxime Injection=Ca-Chl-Monos.pdf) DEFINITION Ceftizoxime Injection is a sterile solution of Ceftizoxime Sodium in a diluent containing one or more tonicity-adjusting agents in Water for Injection. It contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amount of ceftizoxime (C13H13N5O5S2).
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Ceftizoxime / Official Monographs
USP 32
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.0-mm 30-cm; 5- to 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times are about 0.6 for ceftizoxime and 1.0 for salicylic acid.] Suitability requirements Resolution: NLT 4 between the analyte and internal standard peaks Column efficiency: NLT 2000 theoretical plates for the analyte peak Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2% Analysis Samples: Sample solution A or Sample solution B, and Standard solution Calculate the percentage of C13H13N5O5S2 withdrawn from the container, or in the portion of constituted solution taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of ceftizoxime to the internal standard peak from Sample solution A or Sample solution B = peak response ratio of ceftizoxime to the internal RS standard peak from the Standard solution = concentration of ceftizoxime in the Standard CS solution (mg/mL) = nominal concentration of ceftizoxime in Sample CU solution A or Sample solution B (mg/mL) Acceptance criteria: 90.0%115.0% RU PERFORMANCE TESTS INJECTIONS, Constituted Solutions 1: meets the requirements. UNIFORMITY OF DOSAGE UNITS 905: At the time of use, it Meets the requirements
STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791: 5.58.0 PARTICULATE MATTER 788: It meets the requirements for small-volume injections. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements for Injections 1, Labeling. The label states that it is to be thawed just before use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Ceftizoxime RS USP Endotoxin RS
Ceftizoxime for Injection

(Comment on this Monograph)id=m14143=Ceftizoxime for Injection=Ca-Chl-Monos.pdf) DEFINITION Ceftizoxime for Injection contains an amount of Ceftizoxime Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of ceftizoxime (C13H13N5O5S2). ASSAY PROCEDURE Solution A: 1.42 mg/mL of citric acid monohydrate and 1.73 mg/mL of dibasic sodium phosphate in water Solution B: 3.63 mg/mL of monobasic potassium phosphate and 10.73 mg/mL of dibasic sodium phosphate in water Mobile phase: Acetonitrile and Solution A (about 1:9). Filter through a filter (1 m or finer porosity) and adjust the composition, if necessary, to meet the performance requirements under Chromatographic system. Internal standard solution: Dissolve 1.2 g of salicylic acid in 10 mL of methanol and dilute to 200 mL with Solution B. Standard stock solution: 1 mg/mL of USP Ceftizoxime RS in Solution B Standard solution: Transfer 2.0 mL of the Standard stock solution and 5.0 mL of the Internal standard solution to a 100-mL volumetric flask, and dilute with Solution B (0.02 mg/mL of ceftizoxime) to volume. Sample stock solution A: [NOTEPrepare where it is represented as being in a single-dose container.] Constitute Ceftizoxime for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with Solution B to obtain a solution containing 1 mg/mL of ceftizoxime. Sample solution A: Transfer 2.0 mL of Sample stock solution A and 5.0 mL of Internal standard solution to a 100-mL volumetric flask, and dilute with Solution B to volume. Sample stock solution B: [NOTEPrepare where the label states the quantity of ceftizoxime in a given volume of constituted solution.] Constitute Ceftizoxime for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a volume of the constituted solution with Solution B to obtain a solution containing 1 mg/mL of ceftizoxime. Sample solution B: Transfer 2.0 mL of Sample stock solution B and 5.0 mL of Internal standard solution to a 100-mL volumetric flask, and dilute with Solution B to volume.
SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.10 USP Endotoxin Unit/mg of ceftizoxime STERILITY TESTS, Test for Sterility of the Product to be Examined 71: It meets the requirements when tested as directed for Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections INJECTIONS, Labeling 1: Meets the requirements OTHER REQUIREMENTS: It meets the requirements for the Identification tests under Ceftizoxime Sodium. CRYSTALLINITY 695: Meets the requirements PH 791: 6.08.0, in a solution (1 in 10) WATER DETERMINATION, Method I 921: NMT 8.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Ceftizoxime RS USP Endotoxin RS
USP 32
Official Monographs / Ceftriaxone 173

Column efficiency: NLT 1500 theoretical plates, Standard solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C18H18N8O7S3/mg of the Ceftriaxone Sodium taken: Result = (rU/rS) (CS/CU) P
Ceftriaxone Sodium
(Comment on this Monograph)id=m14145=Ceftriaxone Sodium=Ca-Chl-Monos.pdf)
C18H16N8Na2O7S3 31/2H2O 661.60 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2amino-4-thiazolyl)(methoxyimino)acetyl]amino]-8-oxo-3-[ [(1,2,5,6-tetrahydro-2-methyl-5-,6-dioxo-1,2,4-triazin-3yl)thio]methyl]-, disodium salt, [6R-[6,7(Z)]]-, hydrate, (2:7); (6R,7R)-7-[2-(2-Amino-4-thiazolyl)glyoxylamido]-8-oxo-3-[ [(1,2,5,6-tetrahydro-2-methyl-5,6-dioxo-as-triazin-3yl)thio]methyl]-5-thia-1-azabicyclo[4.2.0]-oct-2-ene-2carboxylic acid, 72-(Z)-(O-methyloxime), disodium salt, sesquaterhydrate [104376-79-6]. Anhydrous 598.56 DEFINITION Ceftriaxone Sodium contains the equivalent of NLT 795 g of ceftriaxone (C18H18N8O7S3)/mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak for ceftriaxone from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Sodium 191 ASSAY PROCEDURE Solution A: 13.6 g/L of dibasic potassium phosphate and 4.0 g/L of monobasic potassium phosphate in water Adjust with phosphoric acid or 10 N potassium hydroxide to a pH of 7.0 0.1. Solution B: 25.8 mg/mL of sodium citrate in water Adjust with citric acid solution (1 in 5) to a pH of 5.0 0.1. Mobile phase: Dissolve 3.2 g of tetraheptylammonium bromide in 400 mL of acetonitrile, add 44 mL of Solution A and 4 mL of Solution B, and add water to make 1000 mL. Filter through a membrane filter of 0.5-m or finer porosity, and degas. System suitability solution: 160 g/mL of USP Ceftriaxone Sodium E-Isomer RS and 160 g/mL of USP Ceftriaxone Sodium RS in Mobile phase [NOTEUse this solution promptly after preparation.] Standard solution: 0.2 mg/mL of USP Ceftriaxone Sodium RS in Mobile phase [NOTEUse this solution promptly after preparation.] Sample solution: 0.2 mg/mL of Ceftriaxone Sodium in Mobile phase [NOTEUse this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 4.0-mm 15-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 3 between the ceftriaxone E-isomer and ceftriaxone peaks, System suitability solution
= peak response of ceftriaxone from the Sample solution rS = peak response of ceftriaxone from the Standard solution CS = concentration of USP Ceftriaxone Sodium RS in the Standard solution (mg/mL) CU = concentration of Ceftriaxone Sodium in the Sample solution (mg/mL) P = potency of USP Ceftriaxone Sodium RS in g of C18H18N8O7S3/mg Acceptance criteria: NLT 795 g/mg rU SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 6.08.0, in a solution (1 in 10) WATER DETERMINATION, Method I 921: 8.0%11.0% STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.20 USP Endotoxin Unit/mg of ceftriaxone. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Ceftriaxone Sodium E-Isomer RS USP Ceftriaxone Sodium RS USP Endotoxin RS
Ceftriaxone Injection
(Comment on this Monograph)id=m14147=Ceftriaxone Injection=Ca-Chl-Monos.pdf) DEFINITION Ceftriaxone Injection is a sterile solution of Ceftriaxone Sodium in a diluent containing one or more tonicity-adjusting agents in Water for Injection. It contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amount of ceftriaxone (C18H18N8O7S3). IDENTIFICATION The retention time of the major peak for ceftriaxone from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solvent A: 13.6 mg/mL of dibasic potassium phosphate and 4.0 mg/mL of monobasic potassium phosphate in water Adjust this solution with phosphoric acid or 10 N potassium hydroxide to a pH of 7.0 0.1. Solvent B: 25.8 mg/mL of sodium citrate in water Adjust with citric acid solution (1 in 5) to a pH of 5.0 0.1.
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Ceftriaxone / Official Monographs

USP Ceftriaxone Sodium E-Isomer RS USP Endotoxin RS
USP 32
Mobile phase: Dissolve 3.2 g of tetraheptylammonium bromide in 400 mL of acetonitrile, add 44 mL of Solvent A, 4 mL of Solvent B, and water to 1000 mL. Filter through a membrane filter of 0.5 m or finer porosity. Make adjustments if necessary. System suitability solution: 160 g/mL of USP Ceftriaxone Sodium E-Isomer RS and 160 g/mL of USP Ceftriaxone Sodium RS in Mobile phase. [NOTEUse this solution promptly after preparation.] Standard solution: 0.2 mg/mL of USP Ceftriaxone Sodium RS in Mobile phase [NOTEUse this solution promptly after preparation.] Sample solution: Allow one container of Injection to thaw and mix. Transfer a volume of Injection equivalent to 40 mg of ceftriaxone to a 200-mL volumetric flask, and dilute to volume with Mobile phase. [NOTEUse this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 4.0-mm 15-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 3 between the ceftriaxone E-isomer and ceftriaxone peaks, System suitability solution Column efficiency: NLT 1500 theoretical plates, Standard solution Tailing factor: NMT 2, Standard solution Relative standard deviation: NMT 2%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of ceftriaxone (C18H18N8O7S3) in the Injection taken: Result = (rU/rS) (CS/CU) P 100 = peak response of ceftriaxone from the Sample solution = peak response of ceftriaxone from the Standard rS solution = concentration of USP Ceftriaxone Sodium RS in CS the Standard solution (mg/mL) = nominal concentration of ceftriaxone in the CU Sample solution (g/mL) P = designated potency of ceftriaxone in the USP Ceftriaxone Sodium RS (g/mg) Acceptance criteria: 90.0%115.0% rU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Unit/mg of ceftriaxone STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration. PH 791: 6.08.0 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements under Injections 1, Labeling . The label states that it is to be thawed just prior to use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Ceftriaxone Sodium RS
Ceftriaxone for Injection

(Comment on this Monograph)id=m14149=Ceftriaxone for Injection=Ca-Chl-Monos.pdf) DEFINITION Ceftriaxone for Injection contains an amount of Ceftriaxone Sodium equivalent to NLT 776 g of ceftriaxone (C18H18N8O7S3) per mg, calculated on the anhydrous basis, and the equivalent of NLT 90.0% and NMT 115.0% of the labeled amount of ceftriaxone (C18H18N8O7S3). ASSAY PROCEDURE Solvent A: 13.6 mg/mL of dibasic potassium phosphate and 4.0 mg/mL of monobasic potassium phosphate in water Adjust this solution with phosphoric acid or 10 N potassium hydroxide to a pH of 7.0 0.1. Solvent B: 25.8 mg/mL of sodium citrate in water Adjust with citric acid solution (1 in 5) to a pH of 5.0 0.1. Mobile phase: Dissolve 3.2 g of tetraheptylammonium bromide in 400 mL of acetonitrile, add 44 mL of Solvent A, 4 mL of Solvent B, and water to 1000 mL. Filter through a membrane filter of 0.5 m or finer porosity. Make adjustments if necessary. System suitability solution: 160 g/mL of USP Ceftriaxone Sodium E-Isomer RS and 160 g/mL of USP Ceftriaxone Sodium RS in Mobile phase. [NOTEUse this solution promptly after preparation.] Standard solution: 0.2 mg/mL of USP Ceftriaxone Sodium RS in Mobile phase [NOTEUse this solution promptly after preparation.] Sample solution A: 0.2 mg/mL of Ceftriaxone for Injection in Mobile phase [NOTEUse this solution promptly after preparation.] Sample solution B: [NOTEPrepare where it is represented as being in a single-dose container.] Constitute Ceftriaxone for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents using a suitable hypodermic needle and syringe, and dilute with Mobile phase to obtain a solution containing 180 g/mL of ceftriaxone. [NOTEUse this solution promptly after preparation.] Sample solution C: [NOTEPrepare where the label states the quantity of ceftriaxone in a given volume of constituted solution.] Constitute Ceftriaxone for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a measured volume of the constituted solution with Mobile phase to obtain a solution containing 180 g/mL of ceftriaxone. [NOTEUse this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 4.0-mm 15-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 3 between the ceftriaxone E-isomer and ceftriaxone peaks, System suitability solution Column efficiency: NLT 1500 theoretical plates, Standard solution
USP 32
Tailing factor: NMT 2, Standard solution Relative standard deviation: NMT 2%, Standard solution Analysis Samples: Standard solution and Sample solution A, Sample solution B, or Sample solution C Calculate the quantity, in g/mg, of C18H18N8O7S3 in the portion of Ceftriaxone for Injection taken: Result = (rU/rS) [(CS P)/CU] = peak response of ceftriaxone from Sample solution A rS = peak response of ceftriaxone from the Standard solution CS = concentration of USP Ceftriaxone Sodium RS in the Standard solution (mg/mL) P = designated potency of ceftriaxone in USP Ceftriaxone Sodium RS (g/mg) CU = concentration of Ceftriaxone for Injection in Sample solution A (mg/mL) Calculate the percentage of C18H18N8O7S3 withdrawn from the container, or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) P 100 = peak response of ceftriaxone from the Sample solution = peak response of ceftriaxone from the Standard rS solution = concentration of USP Ceftriaxone Sodium RS in CS the Standard solution (mg/mL) = nominal concentration of ceftriaxone in Sample CU solution B or Sample solution C based on the labeled quantity in the container or in the portion of constituted solution taken, and the extent of dilution (g/mL) P = stated content of ceftriaxone in USP Ceftriaxone Sodium RS (g/mg) Acceptance criteria: Contains an amount of Ceftriaxone Sodium equivalent to NLT 776 g of ceftriaxone per mg, calculated on the anhydrous basis, and 90.0%115.0% of the labeled amount of C18H18N8O7S3. SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Constituted Solutions under Injections 1, Labeling. BACTERIAL ENDOTOXINS 85: NMT 0.20 USP Endotoxin Unit/mg of ceftriaxone STERILITY 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections CRYSTALLINITY 695: Meets the requirements PH 791: 6.08.0, in a solution (1 in 10) WATER DETERMINATION, Method I 921: 8.0%11.0% OTHER REQUIREMENTS: It responds to the Identification tests under Ceftriaxone Sodium. It also meets the requirements for Uniformity of Dosage Units 905 and for Injections 1, Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Ceftriaxone Sodium RS USP Ceftriaxone Sodium E-Isomer RS USP Endotoxin RS rU rU
Official Monographs / Cefuroxime 175
Cefuroxime Axetil
(Comment on this Monograph)id=m14152=Cefuroxime Axetil=Ca-Chl-Monos.pdf)
C20H22N4O10S 510.47 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[[(aminocarbonyl)oxy]methyl]-7-[[2furanyl(methoxyimino)acetyl]amino]-8-oxo-, 1(acetyloxy)ethyl ester, [6R-[67(Z)]]-; (RS)-1-Hydroxyethyl (6R,7R)-7-[2-(2-furyl)glyoxylamido]-3(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene 2carboxylate, 72-(Z)-(O-methyloxime), 1-acetate 3-carbamate [64544-07-6]. DEFINITION Cefuroxime Axetil is a mixture of the diastereoisomers of C20H22N4O10S. It contains the equivalent of NLT 745 g and NMT 875 g of C16H16N4O8S/mg, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197K: Meets the requirements
ASSAY PROCEDURE Solvent A: 0.2M monobasic ammonium phosphate Mobile phase: Methanol and Solvent A (19:31), filtered Internal standard solution: 5.4 mg/mL of acetanilide in methanol System suitability stock solution: 0.16 mg/mL of USP Cefuroxime Axetil Delta-3 Isomers RS in methanol Standard stock solution: 1.2 mg/mL of USP Cefuroxime Axetil RS in methanol System suitability solution: 10.0 mL of Standard stock solution, 5.0 mL of Internal standard solution, and 3.8 mL of System suitability stock solution made to 50.0 mL with Solvent A Standard solution: 10.0 mL of Standard stock solution, 5.0 mL of Internal standard solution, and 3.8 mL of methanol made to 50.0 mL with Solvent A [NOTEUse this Standard solution promptly, or refrigerate and use on the day prepared.] Sample stock solution: 1.2 mg/mL of Cefuroxime Axetil in methanol Sample solution: 10.0 mL of Sample stock solution, 5.0 mL of Internal standard solution, and 3.8 mL of methanol made to 50.0 mL with Solvent A [NOTEUse this Sample solution promptly, or refrigerate and use on the day prepared.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 4.6-mm 25-cm; 5-m packing L13 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for acetanilide, cefuroxime axetil diastereoisomer B, cefuroxime axetil diastereoisomer A, and cefuroxime axetil delta-3 isomers are 0.4, 0.8, 0.9, and 1.0, respectively.]
176
Cefuroxime / Official Monographs
USP 32
Suitability requirements Resolution: NLT 1.5 between cefuroxime axetil diastereoisomer A and B; NLT 1.5 between cefuroxime axetil diastereoisomer A and cefuroxime axetil delta-3 isomers, System suitability solution Column efficiency: NLT 3000 theoretical plates when measured using the cefuroxime axetil diastereoisomer A peak, Standard solution Relative standard deviation: NMT 2.0%, Sample solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C16H16N4O8S in each mg of Cefuroxime Axetil taken: Result = (RU/RS) (CS/CU) (PS/100) (100 K) = ratio of the sum of the peak responses of cefuroxime axetil diastereoisomers A and B to the peak response of the internal standard from the Sample solution = ratio of the sum of the peak responses of RS cefuroxime axetil diastereoisomers A and B to the peak response of the internal standard from the Standard solution = concentration of USP Cefuroxime Axetil RS in CS the Standard solution (mg/mL) = concentration of Cefuroxime Axetil in the Sample CU solution (mg/mL) = designated C16H16N4O8S content of anhydrous PS USP Cefuroxime Axetil RS (g/mg) K = percentage water content of USP Cefuroxime Axetil RS Acceptance criteria: 745875 g/mg SPECIFIC TESTS CRYSTALLINITY 695: Particles that do not show birefringence or exhibit extinction positions are amorphous, and particles that show birefringence and exhibit extinction positions are crystalline. WATER DETERMINATION, Method I 921: NMT 1.5% DIASTEREOISOMER RATIO: Solvent A, Mobile phase, Internal standard solution, System suitability solution, System suitability stock solution, Standard stock solution, Standard solution, Sample stock solution, Sample solution, and Chromatographic system: Prepare as directed in the Assay. Analysis: Proceed as directed in the Assay. Calculate the ratio of cefuroxime axetil diastereoisomer A to the sum of the cefuroxime axetil diastereoisomers A and B. Result = rA/(rA + rB) = peak response of cefuroxime axetil diastereoisomers A rB = peak response of cefuroxime axetil diastereoisomers B Acceptance criteria: 0.480.55 rA ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate whether it is amorphous or crystalline. USP REFERENCE STANDARDS 11 USP Cefuroxime Axetil RS USP Cefuroxime Axetil Delta-3 Isomers RS RU
Cefuroxime Axetil for Oral Suspension

(Comment on this Monograph)id=m14153=Cefuroxime Axetil for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cefuroxime Axetil for Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of cefuroxime (C16H16N4O8S). IDENTIFICATION The retention times of the major peaks for cefuroxime axetil diastereoisomers A and B of the Sample solution correspond to those of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Solution A: 23.0 mg/mL of monobasic ammonium phosphate in water Mobile phase: Methanol and Solution A (19:31) System suitability stock solution A: 1.2 mg/mL of USP Cefuroxime Axetil RS in methanol System suitability stock solution B: 0.16 mg/mL of USP Cefuroxime Axetil Delta-3 Isomers RS in methanol System suitability solution: Transfer 10.0 mL of System suitability stock solution A to a 50-mL volumetric flask. Add 5.0 mL of methanol and 3.8 mL of System suitability stock solution B. Dilute with Solution A to volume. Standard stock solution: 1.2 mg/mL of USP Cefuroxime Axetil RS in methanol Standard solution: Transfer 10.0 mL of Standard stock solution to a 50-mL volumetric flask, add 8.8 mL of methanol, and dilute with Solution A to volume. [NOTEUse this Standard solution promptly, or refrigerate and use on the day prepared.] Sample stock solution: Transfer to a 100-mL volumetric flask a portion equivalent to 250 mg of cefuroxime from Cefuroxime Axetil for Oral Suspension, freshly mixed and free from air bubbles, constituted as directed in the labeling in 50 mL of methanol, and shake by mechanical means for 10 min. Dilute with methanol to volume, and mix. Filter a portion of this stock solution. Sample solution: Transfer 5.0 mL of the filtered Sample stock solution to a 50-mL volumetric flask. Add 13.8 mL of methanol, dilute to volume with Solution A. [NOTEProtect this Sample solution from light and use promptly, or refrigerate and use on the day prepared.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 4.6-mm 25-cm; 5-m packing L13 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for acetanilide, cefuroxime axetil diastereoisomer B, cefuroxime axetil diastereoisomer A, and cefuroxime axetil delta-3 isomers are 0.4, 0.8, 0.9, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between cefuroxime axetil diastereoisomer A and B; NLT 1.5 between cefuroxime axetil diastereoisomer A and cefuroxime axetil delta-3 isomers, System suitability solution
USP 32
Column efficiency: NLT 3000 theoretical plates when measured using the cefuroxime axetil diastereoisomer A peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H16N4O8S in each mL taken: Result = (RU/RS) (CS/CU) PS F (100 K) = sum of the peak responses of the cefuroxime axetil diastereoisomers A and B from the Sample solution = sum of the of the peak responses of the RS cefuroxime axetil diastereoisomers A and B from the Standard solution = concentration of the Standard solution (mg/mL) CS = nominal concentration of cefuroxime axetil in CU the Sample solution (mg/mL) = designated cefuroxime content of anhydrous PS USP Cefuroxime Axetil RS (g/mg) F = conversion correction factor (0.001 mg/g) K = percentage of water content of USP Cefuroxime Axetil RS Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.07 M pH 7.0 phosphate buffer, prepared by dissolving 3.7 mg/mL of monobasic sodium phosphate and 5.7 mg/mL of anhydrous dibasic sodium phosphate in water; 900 mL Apparatus 2: 50 rpm Time: 30 min Analysis: Test 5.0 mL of constituted Cefuroxime Axetil for Oral Suspension equivalent to 125 or 250 mg of cefuroxime. Determine the amount of cefuroxime equivalent dissolved by using UV absorption at the wavelength of maximum absorbance at 280 nm on filtered portions of the solution under test, suitably diluted with Dissolution Medium, if necessary, in comparison with a Standard solution having a known concentration of USP Cefuroxime Axetil RS in the same Medium. Tolerances: NLT 60% (Q) of the labeled amount of C16H16N4O8S UNIFORMITY OF DOSAGE UNITS 905 Constitute Cefuroxime Axetil for Oral Suspension as directed in the labeling for a solid packaged in single-unit containers. Mix, and allow the contents of the container to drain into a beaker for 5 s. Withdraw and assay 5.0 mL of Cefuroxime Axetil for Oral Suspension from the beaker, or the total amount if it is less than 5 mL. It meets the requirements. SPECIFIC TESTS PH 791: 3.57.0 DELIVERABLE VOLUME 698 For multiple-unit containers: Constitute Cefuroxime Axetil for Oral Suspension as directed in the labeling for a solid packaged in multiple-unit containers. It meets the requirements. WATER DETERMINATION, Method I 921: NMT 6.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Cefuroxime Axetil RS USP Cefuroxime Axetil Delta-3 Isomers RS
Cefuroxime Axetil Tablets

(Comment on this Monograph)id=m14154=Cefuroxime Axetil Tablets=Ca-Chl-Monos.pdf) DEFINITION Cefuroxime Axetil Tablets contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of C16H16N4O8S. IDENTIFICATION The retention times of the major peaks for cefuroxime axetil diastereoisomers A and B of the Sample solution correspond to those in the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Solution A: 23.0 mg/mL of monobasic ammonium phosphate in water Mobile phase: Methanol and Solution A (19:31), filtered Internal standard solution: 5.4 mg/mL of acetanilide in methanol System suitability stock solution A: 1.2 mg/mL of USP Cefuroxime Axetil RS in methanol System suitability stock solution B: 0.16 mg/mL of USP Cefuroxime Axetil Delta-3 Isomers RS in methanol System suitability solution: In a 50-mL volumetric flask, mix 10.0 mL of System suitability stock solution A, 5.0 mL of Internal standard solution, and 3.8 mL of System suitability stock solution B. Dilute with Solution A to volume. Standard stock solution: 1.2 mg/mL of USP Cefuroxime Axetil RS in methanol Standard solution: Transfer 10.0 mL of Standard stock solution to a 50-mL volumetric flask, add 5.0 mL of Internal standard solution and 3.8 mL of methanol, and dilute with Solution A to volume. [NOTEUse this Standard solution promptly, or refrigerate and use on the day prepared.] Sample stock solution: Finely powder NLT 10 Tablets. Transfer the powder with the aid of methanol to a volumetric flask of such capacity that when filled to volume, the solution will contain the equivalent of about 2 mg/mL of C16H16N4O8S. Add methanol to fill the volumetric flask to about half its capacity, and shake by mechanical means for about 10 min. Dilute with methanol to volume. Filter a portion of this stock mixture. Sample solution: Transfer 5.0 mL of the filtrate from the Sample stock soluton to a 50-mL volumetric flask. Add 5.0 mL of Internal standard solution and 8.8 mL of methanol, and dilute with Solution A to volume. [NOTEUse this Sample solution promptly, or refrigerate and use on the day prepared.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 4.6-mm 25-cm; 5-m packing L13 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative tretention times for acetanilide, cefuroxime axetil diastereoisomer B, cefuroxime axetil diastereoisomer A, and cefuroxime axetil delta-3 isomers are 0.4, 0.8, 0.9, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between cefuroxime axetil diastereoisomer A and B; NLT 1.5 between cefuroxime axetil diastereoisomer A and cefuroxime axetil delta-3 isomers, System suitability solution
178

USP Cefuroxime Axetil Delta-3 Isomers RS
USP 32
Column efficiency: NLT 3000 theoretical plates when measured using the cefuroxime axetil diastereoisomer A peak, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H16N4O8S in each Tablet: Result = (RU/RS) (CS/CU) (PS) (100 K) = ratios of the sum of the peak responses of the cefuroxime axetil diastereoisomers A and B to the peak responses of the Internal standard of the Sample solution RS = ratios of the sum of the peak responses of the cefuroxime axetil diastereoisomers A and B to the peak responses of the Internal standard of the Standard solution CS = concentration of USP Cefuroxime Axetil RS in the Standard solution (mg/mL) CU = nominal concentration of cefuroxime axetil in the Sample solution (mg/mL) PS = designated C16H16N4O8S content of anhydrous USP Cefuroxime Axetil RS (g/mg) K = percentage of water content of USP Cefuroxime Axetil RS Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1 Medium: 0.07 N hydrochloric acid; 900 mL Apparatus 2: 55 rpm Time: 15 and 45 min Analysis: Determine the amount of C16H16N4O8S dissolved by employing UV absorption at the wavelength of maximum absorbance at 278 nm on filtered portions of the solution under test, suitably diluted with Dissolution Medium, if necessary, in comparison with a Standard solution having a known concentration of USP Cefuroxime Axetil RS, equivalent to 0.01 to 0.02 mg of C16H16N4O8S/mL in the same Medium. Acceptance criteria: NLT 60% (Q) of the labeled amount of C16H16N4O8S is dissolved in 15 min, and NLT 75% (Q) is dissolved in 45 min; except that where Tablets are labeled to contain the equivalent of 500 mg of cefuroxime, NLT 50% (Q) of the labeled amount of C16H16N4O8S is dissolved in 15 min, and NLT 70% (Q) is dissolved in 45 min. Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Apparatus 2: 100 rpm Medium, Times, and Analysis: Proceed as directed under Test 1. Acceptance criteria: NLT 60% (Q) of the labeled amount of C16H16N4O8S is dissolved in 15 min, and NLT 75% (Q) of the labeled amount of C16H16N4O8S is dissolved in 45 min. UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 6.0% RU
Cefuroxime Sodium
(Comment on this Monograph)id=m14155=Cefuroxime Sodium=Ca-Chl-Monos.pdf)
C16H15N4NaO8S 446.37 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[ [(aminocarbonyl)oxy]methyl]-7-[[2furanyl(methoxyimino)acetyl]amino]-8-oxo-, monosodium salt [6R-[6,7 (Z)]]-; Sodium (6R, 7R)-7-[2-(2-furyl)glyoxylamido]-3(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylate, 72-(Z)-(O-methyloxime), carbamate (ester) [56238-63-2] DEFINITION Cefuroxime Sodium contains the equivalent of NLT 855 g and NMT 1000 g of cefuroxime (C16H16N4O8S), calculated on the anhydrous basis. IDENTIFICATION A. The retention time of the major peak for cefuroxime from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191: It meets the requirements. ASSAY PROCEDURE Solution A: pH 3.4 acetate buffer Transfer 50 mL of 0.1 M sodium acetate to a 1000-mL volumetric flask, and dilute with 0.1 N acetic acid to volume. Mobile phase: Acetonitrile and Solution A (1:10) Filter through a membrane filter of 1-m or finer porosity. Internal standard solution: 1.5 mg/mL of orcinol in water Standard solution: Prepare a 1 mg/mL of C16H16N4O8S from USP Cefuroxime Sodium RS in water. Immediately transfer 5.0 mL of the resulting solution to a 100-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with water to volume. This Standard solution contains 0.05 mg of cefuroxime/mL. Sample solution: Prepare 1 mg/mL of Cefuroxime Sodium in water. Immediately transfer 5.0 mL of the resulting solution to a 100-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L15 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for cefuroxime and orcinol are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.5 between the analyte and internal standard peaks in the Standard solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: The labeling indicates whether the Tablets contain amorphous or crystalline Cefuroxime Axetil. If Tablets contain a mixture of amorphous and crystalline Cefuroxime Axetil, label to indicate the percentage of each contained therein. When more than one Dissolution test is given, the labeling states the Dissolution test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Cefuroxime Axetil RS
USP 32
Column efficiency: NLT 1300 theoretical plates in the Standard solution Tailing factor: NMT 2.0 in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of cefuroxime per mg in the portion of Cefuroxime Sodium taken: Result = (RU/RS) (CS/CU) F = peak response ratio of cefuroxime to the internal standard peak from the Sample solution = peak response ratio of cefuroxime to the internal RS standard peak from the Standard solution = concentration of USP Cefuroxime Sodium RS in CS the Standard solution (mg/mL) = concentration of Cefuroxime Sodium in the CU Sample solution (mg/mL) F = conversion factor, 1000 g/mg Acceptance criteria: 8551000 g/mg RU SPECIFIC TESTS PH 791: 6.08.5, in a solution (1 in 10) WATER DETERMINATION, Method I 921: NMT 3.5% STERILITY TESTS 71: Where the label states that Cefuroxime Sodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined , Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85 Where the label states that Cefuroxime Sodium is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.10 USP Endotoxin Unit/mg of cefuroxime. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile, or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cefuroxime Sodium RS USP Endotoxin RS

Internal standard solution: 1.5 mg/mL of orcinol in water Standard solution: 1 mg/mL of C16H16N4O8S from USP Cefuroxime Sodium RS in water Immediately transfer 5.0 mL of the resulting solution to a 100-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with water to volume. Sample solution: Allow a container of Injection to thaw, and mix the solution. Transfer a measured volume of Injection, equivalent to about 50 mg of cefuroxime, to a 50mL volumetric flask, and dilute with water to volume. Immediately transfer 5.0 mL of the resulting solution to a 100-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L15 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for cefuroxime and orcinol are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.5 between the analyte and internal standard peaks, Standard solution Column efficiency: NLT 1300 theoretical plates, Standard solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Sample: Sample solution and Standard solution Calculate the percentage of C16H16N4O8S in each mL of the Injection: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cefuroxime to the internal standard peak from the Sample solution = peak response ratio of cefuroxime to the internal RS standard peak from the Standard solution = concentration of USP Cefuroxime Sodium RS in CS the Standard solution (mg/mL) = nominal concentration of cefuroxime taken to CU prepare the Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% RU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Cefuroxime Injection
(Comment on this Monograph)id=m14156=Cefuroxime Injection=Ca-Chl-Monos.pdf) DEFINITION Cefuroxime Injection is a sterile isoosmotic solution of Cefuroxime Sodium in Water for Injection. It contains one or more suitable buffers and a tonicity-adjusting agent. It contains NLT 90.0% and NMT 120.0% of the labeled amount of cefuroxime (C16H16N4O8S). IDENTIFICATION The retention time of the major peak for cefuroxime from the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Transfer 50 mL of 0.1 M sodium acetate to a 1000-mL volumetric flask, and dilute with 0.1 N acetic acid to volume. Mobile phase: Acetonitrile and Solution A (1:10) Filter through a membrane filter of 1 m or finer porosity.
SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.10 USP Endotoxin Unit/mg of cefuroxime STERILITY TESTS 71: Meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration PH 791: 5.07.5 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Packaging, Containers for Injections. Maintain in the frozen state. LABELING: It meets the requirements under Injections 1, Labels and Labeling. The label states that it is to be thawed just before use, describes conditions for proper storage of
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Calculate the percentage of C16H16N4O8S withdrawn from the container, or in the portion of constituted solution or suspension taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the cefuroxime peak to the internal standard from the Sample solution RS = peak response ratio of the cefuroxime peak to the internal standard from the Standard solution = concentration of cefuroxime in the Standard CS solution (mg/mL) = nominal concentration of cefuroxime, of Sample CU solution A or Sample solution B, based on the labeled quantity in the container or in the portion of constituted solution or suspension taken, and the extent of dilution (mg/mL) [NOTEWhere the test for Uniformity of Dosage Units has been performed using the Analysis for content uniformity, use the average of these determinations as the Assay value.] Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Procedure for content uniformity: Perform the Assay on individual containers using Sample solution A or Sample solution B, or both, as appropriate. SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, the constituted solution for intravenous administration prepared from Cefuroxime for Injection meets the requirements for Constituted Solutions under Injections 1, Labeling. BACTERIAL ENDOTOXINS TEST 85: NMT 0.10 USP Endotoxin Unit/mg of cefuroxime STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 6.08.5, in a solution (1 in 10) WATER DETERMINATION, Method I 921: NMT 3.5% OTHER REQUIREMENTS: Meets the requirements for Injections 1, Labeling and meets the requirements of the Identification tests under Cefuroxime Sodium. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Cefuroxime Sodium RS USP Endotoxin RS RU
the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cefuroxime Sodium RS USP Endotoxin RS
Cefuroxime for Injection

(Comment on this Monograph)id=m14158=Cefuroxime for Injection=Ca-Chl-Monos.pdf) DEFINITION Cefuroxime for Injection contains an amount of Cefuroxime Sodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cefuroxime (C16H16N4O8S). ASSAY PROCEDURE Solution A: Transfer 50 mL of 0.1 M sodium acetate to a 1000-mL volumetric flask, and dilute with 0.1 N acetic acid to volume. Mobile phase: Acetonitrile and Solution A (1:10) Pass through a membrane filter of 1 m or finer porosity. Internal standard solution: 1.5 mg/mL of orcinol in water Standard stock solution: Prepare a solution of USP Cefuroxime Sodium RS equivalent to 1 mg/mL of cefuroxime. Immediately transfer 5.0 mL of the resulting solution to a 100-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with water to volume. Sample solution A (where it is represented as being in a single-dose container): Constitute Cefuroxime for Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute quantitatively with water to obtain a solution containing about 1 mg/mL of cefuroxime. Immediately transfer 5.0 mL of the resulting solution to a 100-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with water to volume. Sample solution B (where the label states the quantity of cefuroxime in a given volume of constituted solution or suspension): Constitute Cefuroxime for Injection in a volume of water, corresponding to the volume of solvent specified in the labeling. Dilute a measured volume of the constituted solution or suspension quantitatively with water to obtain a solution containing about 1 mg/mL of cefuroxime. Immediately transfer 5.0 mL of the resulting solution to a 100-mL volumetric flask, add 20.0 mL of Internal standard solution, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; 5-m packing L15 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times are 0.5 for cefuroxime and 1.0 for orcinol.] Suitability requirements Resolution: NLT 3.5 between the analyte and internal standard peaks in the Standard solution Column efficiency: NLT 1300 theoretical plates, Standard solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Sample solution A, or Sample solution B, and Standard solution
Oxidized Cellulose
(Comment on this Monograph)id=m14200=Oxidized Cellulose=Ca-Chl-Monos.pdf) DEFINITION Oxidized Cellulose contains NLT 16.0% and NMT 24.0% of carboxyl groups (COOH), calculated on the dried basis. It is sterile. IDENTIFICATION PROCEDURE Sample solution: 200 mg in 10 mL of 0.25 N sodium hydroxide Analysis: Shake the Sample solution for 1 min. Add 10 mL of water, and shake: the solution so obtained shows no more than a slight haze, and is substantially free from fibers
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and foreign particles. Allow to stand for 10 min: any swollen fibers initially present are no longer visible. Acidify with 3 N hydrochloric acid. Acceptance criteria: A flocculent white precipitate is formed. ASSAY PROCEDURE Sample solution: 500 mg of Oxidized Cellulose, previously dried over phosphorus pentoxide in vacuum for 18 h, in a 125-mL conical flask Add 50.0 mL of calcium acetate solution (1 in 50), swirl until the sample is completely covered, allow the mixture to stand for 30 min, then add phenolphthalein TS. Analysis: Titrate the solution with 0.1 N sodium hydroxide VS. Perform a blank determination by titrating 50.0 mL of the calcium acetate solution, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N sodium hydroxide is equivalent to 4.502 mg of COOH. Acceptance criteria: 16.0%24.0% IMPURITIES Inorganic Impurities LIMIT OF NITROGEN Sample: 1 g Analysis: Previously dried sample in vacuum over phosphorus pentoxide for 18 h, in a 500-mL Kjeldahl flask Arrange a 125-mL conical flask, containing 30 mL of boric acid solution (1 in 25) and 6 drops of mixed indicator (1 part of methyl red TS and 4 parts of bromocresol green TS), beneath the condenser of the distillation apparatus so that the tip of the condenser is well below the surface of the boric acid solution. To the Kjeldahl flask containing the Sample, add 1 g of Devardas alloy, 100 mL of recently boiled water, a small lump of paraffin, and 100 mL of 1 N sodium hydroxide. Connect the Kjeldahl flask to the condenser by a suitable trap bulb. Heat the mixture in the flask until 4550 mL of distillate has collected in the receiver. Rinse the condenser, and titrate the boric acid solution with 0.02 N sulfuric acid VS to a pale pink endpoint. Perform a blank determination, and make any necessary correction. Each mL of 0.02 N sulfuric acid is equivalent to 0.2801 mg of nitrogen. Acceptance criteria: NMT 0.5% RESIDUE ON IGNITION 281: NMT 0.15% Organic Impurities LIMIT OF FORMALDEHYDE Sample: 500 mg Analysis: Transfer the Sample to a 500-mL iodine flask. Add 250 mL of water, and allow to stand for NLT 2 h with intermittent shaking. Pipet 0.50 mL of the supernatant into a glass-stoppered test tube, and add 10 mL of chromotropic acid TS. Stopper the tube loosely, and heat in a boiling water bath for 30 min. Cool, and determine the absorbance of the solution at 570 nm with a suitable spectrophotometer, using a mixture of 0.5 mL of water and 10 mL of chromotropic acid TS as the blank. Acceptance criteria: The absorbance does not exceed that produced when 0.50 mL of dilute formaldehyde solution (1 in 40,000) is treated in the same manner (0.5%). SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements, the test specimen weighing approximately 250 mg, and 0.5 mL of 0.1 N sodium hydroxide being added to the portions of media used LOSS ON DRYING 731: Dry in vacuum over phosphorus pentoxide for 18 h: it loses NMT 15.0% of its weight.
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids, protected from direct sunlight. Store in a cold place. LABELING: The package bears a statement to the effect that the sterility of Oxidized Cellulose cannot be guaranteed if the package bears evidence of damage, or if the package has been previously opened. Oxidized Cellulose meets the requirements for Injections 1, Labeling.
Oxidized Regenerated Cellulose

(Comment on this Monograph)id=m14210=Oxidized Regenerated Cellulose=Ca-Chl-Monos.pdf) DEFINITION Oxidized Regenerated Cellulose contains NLT 18.0% and NMT 24.0% of carboxyl groups (COOH), calculated on the dried basis. It is sterile. IDENTIFICATION PROCEDURE Sample solution: 200 mg in 10 mL of 0.25 N sodium hydroxide Analysis: Shake the Sample solution for 1 min. Add 10 mL of water, and shake: the solution so obtained shows no more than a slight haze and is substantially free from fibers and from foreign particles. Allow to stand for 10 min: any swollen fibers initially present are no longer visible. Acidify with 3 N hydrochloric acid. Acceptance criteria: A flocculent white precipitate is formed. ASSAY PROCEDURE Sample solution: 1 g of Oxidized Regenerated Cellulose, previously dried at 90 for 2 h, in a 250-mL conical flask. Pipet 10 mL of 0.5 N sodium hydroxide VS into the flask, swirl to dissolve, and add 100 mL of water. Analysis: Immediately titrate with 0.1 N hydrochloric acid VS to a phenolphthalein endpoint. Perform a blank determination, and note the difference in volumes required. Each mL of the difference in volumes of 0.1 N hydrochloric acid consumed is equivalent to 4.50 mg of COOH. Acceptance criteria: 18.0%24.0% OTHER COMPONENTS NITROGEN CONTENT Sample: 1 g Analysis: Transfer a previously dried Sample in a 500-mL Kjeldahl flask. Arrange a 125-mL conical flask, containing 30 mL of boric acid solution (1 in 25) and 6 drops of mixed indicator (1 part of methyl red TS and 4 parts of bromocresol green TS), beneath the condenser of the distillation apparatus so that the tip of the condenser is well below the surface of the boric acid solution. To the Kjeldahl flask containing the Sample, add 1 g of Devardas alloy, 100 mL of recently boiled water, a small lump of paraffin, and 100 mL of 1 N sodium hydroxide. Connect the Kjeldahl flask to the condenser by a suitable trap bulb. Heat the mixture in the flask until 4550 mL of distillate has collected in the receiver. Rinse the condenser, and titrate the boric acid solution with 0.02 N sulfuric acid VS to a pale pink endpoint that persists for 30 s. Perform a blank determination, and make any necessary correction. Each mL of 0.02 N sulfuric acid is equivalent to 0.2801 mg of nitrogen.
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NMT 0.5%
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follows. Add 10 mL of 5 N nitric acid, 10.0 mL of ammonium vanadate TS, and about 60 mL of water. Swirl, and add 10.0 mL of a freshly prepared solution of 2.5 g of ammonium molybdate in 50 mL of warm water. Dilute with water to volume. Concomitantly determine the absorbances, AU and AS, of the solutions from the Standard solution and the Sample solution, respectively, at 400 nm with a suitable spectrophotometer, using the reagent blank to set the instrument. Calculate the percentage of total phosphate taken: (AU/AS) (10,000/W) = absorbance of the solution from the Sample solution = absorbance of the solution from the Standard AS solution W = weight of Cellulose Sodium Phosphate taken (mg) Calculate the percentage of inorganic bound phosphate in the undried Cellulose Sodium Phosphate by subtracting from this result the percentage of Free phosphate. Acceptance criteria: 31.0%36.0% OTHER COMPONENTS NITROGEN DETERMINATION, Method I 461: NMT 1.0% FREE PHOSPHATE Standard solution: Prepare as directed under Inorganic Bound Phosphate. Sample solution: 2 g of Cellulose Sodium Phosphate in 100 mL of water, stir, allow to stand for 5 min, stir again, and filter through moderately retentive filter paper, collecting the filtrate in a dry flask. Analysis: Transfer 2.0 mL of the Standard solution and 5.0 mL of the Sample solution to separate 100-mL volumetric flasks. Proceed as directed in the Analysis under Inorganic Bound Phosphate, beginning with Treat each of these. Calculate the percentage of free phosphate taken: (AU/AS) (4,000/W) AU
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.15% Organic Impurities LIMIT OF FORMALDEHYDE Sample: 500 mg Analysis: Transfer the Sample to a 500-mL iodine flask. Add 250 mL of water, and allow to stand for NLT 2 h with intermittent shaking. Pipet 0.5 mL of the supernatant into a glass-stoppered test tube, and add 10 mL of chromotropic acid TS. Stopper the tube loosely, and heat in a boiling water bath for 30 min. Cool, and determine the absorbance of the solution at 570 nm, with a suitable spectrophotometer, using a mixture of 0.5 mL of water and 10 mL of chromotropic acid TS as the blank. Acceptance criteria: The absorbance does not exceed that produced when 0.5 mL of dilute formaldehyde solution (1 in 40,000) is treated in the same manner (0.5% CH2O). SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements; the test specimen weighing approximately 250 mg and 0.5 mL of 0.1 N sodium hydroxide being added to the portions of media used. LOSS ON DRYING 731: Dry about 150 mg at 90 for 2 h: it loses NMT 15% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids, protected from direct sunlight. Store at controlled room temperature. LABELING: The package bears a statement to the effect that the sterility of Oxidized Regenerated Cellulose cannot be guaranteed if the package bears evidence of damage, or if the package has been previously opened. Oxidized Regenerated Cellulose meets the requirements for Injections 1, Labeling.
Cellulose Sodium Phosphate

(Comment on this Monograph)id=m14260=Cellulose Sodium Phosphate=Ca-Chl-Monos.pdf) DEFINITION Cellulose Sodium Phosphate is prepared by phosphorylation of alpha cellulose. It has an inorganic bound phosphate content of NLT 31.0% and NMT 36.0%, calculated on the dried basis. ASSAY INORGANIC BOUND PHOSPHATE Standard solution: Transfer 358.2 mg of monobasic potassium phosphate, primary standard grade, to a 250-mL volumetric flask, and dissolve in and dilute with water to volume. [NOTEEach mL of this solution contains 1.0 mg of phosphate.] Sample solution: Transfer 250 mg of Cellulose Sodium Phosphate to a 250-mL conical flask. Rinse to the bottom with a few mL of water. Add 10 mL of a mixture of 20 mL of perchloric acid and 15 mL of nitric acid. Heat cautiously to the production of dense, white fumes, cool the clear, almost colorless, solution, and transfer to a 100-mL volumetric flask with the aid of water. Dilute with water to volume. Analysis Samples: Standard solution and Sample solution Transfer 2.0-mL portions of the Standard solution and the Sample solution to separate 100-mL volumetric flasks. Treat each of these and a third flask, providing the blank, as
= absorbance of the solution from the Sample solution AS = absorbance of the solution from the Standard solution W = weight of undried Cellulose Sodium Phosphate taken (mg) Acceptance Criteria: NMT 3.5% of free phospate, calculated on the dried basis SODIUM CONTENT Standard stock solution: 508.5 mg of sodium chloride, previously dried at 105 for 2 h, in 100 mL of water Transfer to a 1000-mL volumetric flask, and dilute with water to volume. [NOTEEach mL of this solution contains 200 g of sodium] Standard solutions: Transfer 5.0, 10.0, 15.0, and 20.0 mL of the Standard stock solution to separate 100-mL volumetric flasks, and dilute each with water to volume. Sample solution: 250 mg of Cellulose Sodium Phosphate in 10 mL of a mixture of 20 mL of perchloric acid and 15 mL of nitric acid. Heat cautiously to the production of dense, white fumes, cool, transfer to a 1000-mL volumetric flask, and dilute with water to volume. Analysis: Concomitantly determine the emission of the Sample solution and of each Standard solution at the sodium emission line of 589 nm, with a suitable flame photometer. Plot the emissions of the Standard solutions versus their concentration of sodium, and draw a straight line best fitting the four plotted points. From the graph so obtained, determine the concentration of sodium in the Sample
AU
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solution. Calculate the percentage of sodium in the undried Cellulose Sodium Phosphate. Acceptance criteria: 9.5%13.0%, calculated on the dried basis IMPURITIES Inorganic Impurities HEAVY METALS, Method III 231:
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NMT 40 ppm
Cellulose Sodium Phosphate for Oral Suspension

(Comment on this Monograph)id=m14265=Cellulose Sodium Phosphate for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cellulose Sodium Phosphate for Oral Suspension contains Cellulose Sodium Phosphate. It has an inorganic bound phosphate content of NLT 28.0% and NMT 36.0%, calculated on the dried basis. ASSAY INORGANIC BOUND PHOSPHATE Standard solution: Transfer 358.2 mg of monobasic potassium phosphate, primary standard grade, to a 250-mL volumetric flask, and dissolve in and dilute with water to volume. [NOTEEach mL of this solution contains 1.0 mg of phosphate.] Sample solution: Transfer the equivalent to 250 mg of cellulose sodium phosphate from Cellulose Sodium Phosphate for Oral Suspension to a 250-mL conical flask. Rinse to the bottom with a few mL of water. Add 10 mL of a mixture of 20 mL of perchloric acid and 15 mL of nitric acid. Heat cautiously to the production of dense, white fumes, cool the clear, almost colorless, solution, and transfer to a 100-mL volumetric flask with the aid of water. Dilute with water to volume. Analysis Samples: Standard solution and Sample solution Transfer 2.0-mL portions of the Standard solution and the Sample solution to separate 100-mL volumetric flasks. Treat each of these and a third flask, providing the blank, as follows. Add 10 mL of 5 N nitric acid, 10.0 mL of ammonium vanadate TS, and about 60 mL of water. Swirl, and add 10.0 mL of a freshly prepared solution of 2.5 g of ammonium molybdate in 50 mL of warm water. Dilute with water to volume. Concomitantly determine the absorbances, AU and AS, of the solutions from the Standard solution and the Sample solution, respectively, at 400 nm with a suitable spectrophotometer, using the reagent blank to set the instrument. Calculate the percentage of total phosphate in the portion of Cellulose Sodium Phosphate for Oral Suspension: (AU/AS) (10,000/W) AU = absorbance of the solution from the Sample solution = absorbance of the solution from the Standard AS solution W = weight of Cellulose Sodium Phosphate for Oral Suspension taken (mg) Calculate the percentage of inorganic bound phosphate in the portion of undried Cellulose Sodium Phosphate for Oral
SPECIFIC TESTS PH 791: Place 3 g of it in a 100-mL beaker, add 60 mL of water, and stir occasionally for 5 min. Filter through a sintered-glass crucible. The pH of the filtrate is 6.09.0. LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 10.0% of its weight. CALCIUM BINDING CAPACITY Standard solution: Transfer 0.33 g of dried calcium carbonate, primary standard grade, in a 250-mL beaker with the aid of a few mL of water, and dilute with water to about 50 mL. Carefully and dropwise, add 2 N hydrochloric acid until all of the solid dissolves, and add 2 drops in excess. Heat the solution to boiling, and boil for 5 min. Cool the solution, and transfer to a 1000-mL volumetric flask. Dilute with water to volume. Calculate the molarity, M, of the solution taken: Result = g/100.09 g = weight of calcium carbonate taken (g) Standard edetate disodium titrant: Dissolve 10 g of edetate disodium in 100 mL of water. Slowly add alcohol until the first permanent precipitate is formed. Filter, and discard the solid. Add an equal volume of alcohol to the filtrate. Filter the resulting precipitate, discard the filtrate, and wash the residue on the filter, first with acetone, then with ethyl ether. Dry at 80 for 4 days at about 50% relative humidity. Transfer 3.72 g of this purified edetate disodium to a 1000-mL volumetric flask, and dissolve with water. Dilute with water to volume. Calculate the molarity, MS, of the solution taken: Result = w/372.24 w = weight of the purified edetate disodium taken (g) Analysis: Transfer 0.15 0.02 g of Cellulose Sodium Phosphate to a 250-mL beaker. Add 150.0 mL of the Standard solution, and stir the mixture for 5 min on a magnetic stirrer. Filter, discarding the first few mL of the filtrate. To 50.0 mL of the filtrate, add about 50 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Titrate with Standard edetate disodium titrant to a permanent deep blue color, and designate the number of mL consumed as VS. Calculate the calcium binding capacity of the undried Cellulose Sodium Phosphate, in mmol/g: Result = (150M 3VS MS)/W W = weight of Cellulose Sodium Phosphate taken (g) Acceptance criteria: The calcium binding capacity is NLT 1.8 mmol/g, calculated on the dried basis.
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edetate disodium titrant to a permanent deep blue color, and designate the number of mL consumed as VS. Calculate the calcium binding capacity, in mmol/g, of the portion of undried Cellulose Sodium Phosphate for Oral Suspension: Result = (150M 3VS MS)/W = weight of Cellulose Sodium Phosphate for Oral Suspension taken, and the other terms are as defined above (g) Acceptance criteria: The calcium binding capacity is NLT 1.8 mmol/g, calculated on the dried basis ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store in a refrigerator. W
Suspension by subtracting from this result the percentage of free phosphate. Acceptance criteria: 28.0%36.0% of inorganic bound phosphate, calculated on the dried basis OTHER COMPONENTS FREE PHOSPHATE Standard solution: Prepare as directed under Inorganic Bound Phosphate. Sample solution: Equivalent to 2 g of cellulose sodium phosphate from Cellulose Sodium Phosphate for Oral Suspension in 100 mL of water Stir, allow to stand for 5 min, stir again, and filter through moderately retentive filter paper, collecting the filtrate in a dry flask. Analysis: Transfer 2.0 mL of the Standard solution and 5.0 mL of the Sample solution to separate 100-mL volumetric flasks. Proceed as directed in the Analysis under Inorganic Bound Phosphate, beginning with Treat each of these. Calculate the percentage of free phosphate in the portion of Cellulose Sodium Phosphate for Oral Suspension: (AU/AS) (4000/W) = absorbance of the solution from the Sample solution = absorbance of the solution from the Standard As solution W = weight of undried Cellulose Sodium Phosphate for Oral Suspension taken (mg) Acceptance criteria: NMT 6.0% free phosphate, calculated on the dried basis. AU SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 10.0% of its weight. CALCIUM BINDING CAPACITY Standard solution: Transfer 0.33 g of dried calcium carbonate, primary standard grade, in a 250-mL beaker with the aid of a few mL of water, and dilute with water to about 50 mL. Carefully and dropwise add 2 N hydrochloric acid until all of the solid dissolves, and add 2 drops in excess. Heat the solution to boiling, and boil for 5 min. Cool the solution, and transfer to a 1000-mL volumetric flask. Dilute with water to volume. Calculate the molarity, M, of the solution taken: Result = g/100.09 g = weight of calcium carbonate taken (g) Standard edetate disodium titrant: Dissolve 10 g of edetate disodium in 100 mL of water. Slowly add alcohol until the first permanent precipitate is formed. Filter, and discard the solid. Add an equal volume of alcohol to the filtrate. Filter the resulting precipitate, discard the filtrate, and wash the residue on the filter, first with acetone, then with ethyl ether. Dry at 80 for 4 days at about 50% relative humidity. Transfer about 3.72 g of this purified edetate disodium to a 1000-mL volumetric flask, and dissolve with water. Dilute with water to volume. Calculate the molarity, MS, of the solution taken: Result = w/372.24 w = weight of the purified edetate disodium taken (g) Analysis: Transfer an equivalent to 0.15 g of cellulose sodium phosphate from Oral Suspension, to a 250-mL beaker. Add 150.0 mL of the Standard solution, and stir the mixture for 5 min on a magnetic stirrer. Filter, discarding the first few mL of the filtrate. To 50.0 mL of the filtrate add about 50 mL of water, 15 mL of 1 N sodium hydroxide, and 300 mg of hydroxy naphthol blue. Titrate with Standard
Cephalexin
(Comment on this Monograph)id=m14280=Cephalexin=Ca-ChlMonos.pdf)
365.40 C16H17N3O4S H2O 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7[(aminophenylacetyl)amino]-3-methyl-8-oxo-, monohydrate, [6R-[6,7 (R*)]]-; (6R,7R)-7-[(R)-2-Amino-2-phenylacetamido]-3-methyl-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid monohydrate [23325-78-2] Anhydrous [15686-71-2]. 347.40 DEFINITION Cephalexin has a potency of NLT 950 g and NMT 1030 g of C16H17N3O4S/mg, calculated on the anhydrous basis. IDENTIFICATION A. The IR absorption spectrum of a potassium bromide dispersion of it exhibits maxima only at the same wavelengths as that of a similar solution of USP Cephalexin RS. B. The UV absorption spectrum of a solution (1 in 50,000) exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Cephalexin RS, concomitantly measured, and the absorptivity, calculated on the anhydrous basis, at the wavelength of maximum absorbance at 262 nm is NLT 95.0% and NMT 104.0% of that of USP Cephalexin RS, the potency of the Reference Standard being taken into account. C. THIN-LAYER CHROMATOGRAPHY Standard solution: 25 mg/mL of USP Cephalexin RS in water, with 0.1 N hydrochloric acid Sample solution: 25 mg/mL in water, with 0.1 N hydrochloric acid Chromatographic system See (Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Ethyl acetate, acetonitrile, glacial acetic acid, and water (21:7:7:9)
USP 32
Analysis Samples: Standard solution and Sample solution Allow the spots to dry, place the plate in a saturated chamber containing the solvent system and lined with filter paper. Develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the plate to air-dry, and examine under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Prepare 1015 mL of a suitable mixture of acetonitrile, methanol, triethylamine, and water (100:50:15:850). Dissolve 1.0 g sodium-1-pentanesulfonate in this mixture and adjust with phosphoric acid to a pH of 3.0 0.1. Internal standard stock solution: Dissolve 300 mg of 1hydroxyphenzotriazole in 10 mL of methanol. Internal standard solution: 0.3 mg/mL of 1hydroxyphenzotriazole from Internal standard stock solution diluted with Mobile phase Standard stock solution: 1 mg/mL of USP Cephalexin RS in water Standard solution: Mix 10 mL of Standard stock solution with 15 mL of Internal standard solution. Sample stock solution: 1 mg/mL of Cephalexin in water Sample solution: Mix 10 mL of Standard stock solution with 15 mL of Internal standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for 1hydroxybenzotriazole and cephalexin are about 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the internal standard and the analyte peaks, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C16H17N3O4S per mg of the Cephalexin taken: Result = (RU/RS) (CS/CU) P RU RS CS CU P = ratio of the responses of the cephalexin peak to the 1-hydroxybenzotriazole peak from the Sample solution = ratio of the responses of the cephalexin peak to the 1-hydroxybenzotriazole peak from the Standard solution = concentration of USP Cephalexin RS in the Standard stock solution (mg/mL) = concentration of cephalexin in the Sample stock solution (mg/mL) = designated content of cephalexin in USP Cephalexin RS (g/mg)
Official Monographs / Cephalexin 185

Acceptance criteria: 9501030 g/mg
IMPURITIES Organic Impurities PROCEDURE 1 Solution A: Dissolve 1 g of sodium 1-pentanesulfonate in a mixture of 1000 mL of water and 15 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1. Solution B: Dissolve 1 g of sodium 1-pentanesulfonate in a mixture of 300 mL of water and 15 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1, and add 350 mL of acetonitrile and 350 mL of methanol. Mobile Phase: See the gradient table below.
Time (min) 0 1 33.3 34.3 Solution A (%) 100 100 0 0 Solution B (%) 0 0 100 100
Diluent: 18 mg/mL of monobasic potassium phosphate in water Standard solutions: 0.08 mg/mL and 0.16 mg/mL of C16H17N3O4S from USP Cephalexin RS in Diluent, taking into account the stated potency of the USP Cephalexin RS Sample solution: 5 mg/mL of Cephalexin in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1 mL/min Injection size: 20 L Analysis Samples: Standard solutions and Sample solution Plot the responses of the cephalexin peaks from the Standard solutions versus their concentrations, calculated on the anhydrous basis, in mg/mL, and draw a straight line through the two points and zero. From the line so obtained and the peak responses of the Sample solution, determine the concentration, I, in mg/mL, of each cephalexin-related substance of the Sample solution other than the cephalexin peak. Calculate the percentage of each cephalexin-related substance: Result = I/C 100 = concentration, calculated on the anhydrous basis of Cephalexin taken to prepare the Sample solution (mg/mL) Acceptance criteria Individual impurities: NMT 1.0% of any individual cephalexin-related substance Total impurities: NMT 5.0 % PROCEDURE 2: DIMETHYLANILINE 223: Meets the requirement SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +149 to +158 Sample solution: 5 mg/mL, in pH 4.4 neutralized phthalate buffer (See Reagents, Indicators, and SolutionsBuffer Solutions) C
186
Cephalexin / Official Monographs
USP 32
Standard stock solution: 1 mg/mL of USP Cephalexin RS in water Standard solution: Mix 10 mL of Standard stock solution and 15 mL of Internal standard solution. Sample stock solution: 1.15 mg/mL of Cephalexin Hydrochloride in water Sample solution: Mix 10 mL of Sample stock solution with 15 mL of Internal Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for 1hydroxybenzotriazole and cephalexin are 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the internal standard and the analyte peaks, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C16H17N3O4S in each mg of Cephalexin Hydrochloride taken: Result = (RU/RS) (CS/CU) P = ratio of the responses of the cephalexin peak to the 1-hydroxybenzotriazole peak from the Sample solution RS = ratio of the responses of the cephalexin peak to the 1-hydroxybenzotriazole peak from the Standard solution CS = concentration of USP Cephalexin RS, mg/mL in the Standard stock solution CU = concentration of Cephalexin Hydrochloride from the Sample stock solution (mg/mL) P = stated content of cephalexin in USP Cephalexin RS (g/mg) Acceptance criteria: 800880 g/mg RU IMPURITIES Organic Impurities PROCEDURE 1 Solution A: 1 g of sodium 1-pentanesulfonate in a mixture of 1000 mL of water and 15 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1. Solution B: 1 g of sodium 1-pentanesulfonate in a mixture of 300 mL of water and 15 mL of triethylamine. Adjust with phosphoric acid to a pH of 2.5 0.1, and add 350 mL of acetonitrile and 350 mL of methanol. Mobile phase: See the gradient table below.
Time (min) 0 1 33.3 34.3 Solution A (%) 100 100 0 0 Solution B (%) 0 0 100 100
CRYSTALLINITY 695: Meets the requirements PH 791: 3.05.5, in an aqueous suspension containing 50 mg/mL WATER DETERMINATION, Method I 921: 4.0%8.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cephalexin RS
Cephalexin Hydrochloride
(Comment on this Monograph)id=m14282=Cephalexin Hydrochloride=Ca-Chl-Monos.pdf) 401.87 C16H17N3O4S HCl H2O 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7[(aminophenylacetyl)amino]-3-methyl-8-oxo-, monohydrochloride, monohydrate, [6R-[6,7 (R*)]]-; (6R,7R)-7-[(2R)-2-Amino-2-phenylacetamido]-3-methyl-8-oxo-5thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, monohydrochloride, monohydrate; 7-(D-2-Amino-2-phenylacetamido)-3-methyl-3-cephem-4carboxylic acid hydrochloride monohydrate [105879-42-3]. DEFINITION Cephalexin Hydrochloride contains the equivalent of NLT 800 g and NMT 880 g of cephalexin (C16H17N3O4S) per mg. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 25 mg/mL of USP Cephalexin RS in water with 0.1 N hydrochloric acid Sample solution: 25 mg/mL in water with 0.1 N hydrochloric acid Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Ethyl acetate, acetonitrile, glacial acetic acid, and water (21:7:7:9) Analysis Samples: Standard solution and Sample solution Allow the spots to dry, place the plate in a saturated chamber containing the solvent system and lined with filter paper. Develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the plate to air-dry, and examine under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. The UV absorption spectrum of a solution (1 in 50,000) exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Cephalexin RS, concomitantly measured. C. IDENTIFICATION TESTSGENERAL, Chloride 191: 10 mg/mL meets the requirements for Chloride. ASSAY PROCEDURE Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a mixture of acetonitrile, methanol, triethylamine, and water (100:50:15:850), adjusted with phosphoric acid to a pH of 3.0 0.1 Internal standard solution: 0.3 mg/mL of 1hydroxybenzotriazole dissolved in methanol and diluted with Mobile phase (1:99) to volume.
Diluent: water
18 mg/mL of monobasic potassium phosphate in
USP 32
Standard solutions: 0.08 mg/mL and 0.16 mg/mL of C16H17N3O4S from USP Cephalexin RS in Diluent, taking into account the stated potency of the USP Cephalexin RS Sample solution: 6 mg/mL of Cephalexin Hydrochloride in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1 mL/min Injection size: 20 L Analysis Samples: Standard solutions and Sample solution Plot the responses of the cephalexin peaks of the Standard solutions versus their concentrations, calculated on the anhydrous basis, in mg/mL, and draw a straight line through the two points and zero. From the line so obtained and the peak responses of the Sample solution, determine the concentration, I, in mg/mL, of each cephalexin-related substance from the Sample solution other than the cephalexin peak. Calculate the percentage of each cephalexin-related substance represented by each peak of the Sample solution, other than the cephalexin peak. Result = (I/C) 100 = concentration of each cephalexin-related substance other than cephalexin in the Sample solution (mg/mL) C = concentration mg/mL of cephalexin from the Sample solution Acceptance criteria Individual impurities: NMT 1.0% of any individual cephalexin-related substance is found. Total impurities: NMT 5.0 % PROCEDURE 2: DIMETHYLANILINE 223: Meets the requirement SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 1.53.0, in a solution containing 10 mg/mL WATER DETERMINATION, Method I 921: 3.0%6.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cephalexin RS I

Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1 in 15) of ninhydrin in acetone (60:40:1.5) Analysis Samples: Standard solution and Sample solution Allow the solvent front to move the length of the plate in the Pre-developing solvent system, remove the plate from the chamber, and allow the solvent to evaporate. On this plate apply 10 L each of the Sample solution and Standard solution. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a mixture of acetonitrile, methanol, triethylamine, and water (100:50:15:850), adjusted with phosphoric acid to a pH of 3.0 0.1 Internal standard solution: 0.3 mg/mL of 1hydroxybenzotriazole dissolved in methanol and diluted with Mobile phase (1:99) Standard stock solution: 1 mg/mL of USP Cephalexin RS in water Standard solution: Mix 10 mL of Standard stock solution with 15 mL of Internal standard solution. Sample stock solution: Equivalent to 1 mg/mL of cephalexin from combined contents of NLT 20 Capsules in water. Sonicate, if necessary, to dissolve the cephalexin. Filter, if necessary, to obtain a clear solution. Sample solution: Mix 10 mL of Sample stock solution with 15 mL of Internal standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for 1hydroxybenzotriazole and cephalexin are 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the internal standard and the analyte peaks, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O4S in the portion of Capsules taken: Result = (RU/RS) (CS/CU) P F 100
Cephalexin Capsules
(Comment on this Monograph)id=m14290=Cephalexin Capsules=Ca-Chl-Monos.pdf) DEFINITION Cephalexin Capsules contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cephalexin (C16H17N3O4S). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 3 mg/mL of USP Cephalexin RS in water Sample solution: 3 mg/mL of cephalexin from Capsule in water and filter Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
188
RU
USP 32
Analysis Samples: Standard solution and Sample solution Allow the solvent front to move the length of the plate in the Pre-developing solvent system, remove the plate from the chamber, and allow the solvent to evaporate. On this plate apply 10 L each of the Sample solution and the Standard solution. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution ASSAY PROCEDURE Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a mixture of acetonitrile, methanol, triethylamine, and water (100:50:15:850), adjusted with phosphoric acid to a pH of 3.0 0.1. Standard stock solution: 1 mg/mL of USP Cephalexin RS in water Standard solution: Mix 10 mL of Standard stock solution with 15 mL of Internal standard solution. System suitability stock solution: 0.3 mg/mL of 1hydroxybenzotriazole dissolved in methanol, and diluted with Mobile phase (1:99) System suitability solution: 10 mL of System suitability stock solution and 15 mL of Internal standard solution Sample stock solution: Nominally equivalent to 1 mg/mL of cephalexin from Oral Suspension, constituted as directed in the labeling, freshly mixed and free from air bubbles. Sonicate, if necessary, to assure complete dissolution of the cephalexin. Filter, if necessary, to obtain a clear solution. Sample solution: Mix 10 mL of Sample stock solution and 15 mL of Internal standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution and System suitability solution [NOTEThe relative retention times for 1hydroxybenzotriazole and cephalexin are about 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the 1-hydroxybenzotriazole peak and the cephalexin peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O4S in each mL of Oral Suspension taken: Result = (rU/rS) (CS/CU) P F 100 rU rS CS = cephalexin peak response from the Sample solution = cephalexin peak response from the Standard solution = concentration of USP Cephalexin RS in the Standard stock solution (mg/mL)
= ratio of the response of the cephalexin peak to the 1-hydroxybenzotriazole peak from the Sample solution = ratio of the response of the cephalexin peak to RS the 1-hydroxybenzotriazole peak from the Standard solution = concentration of USP Cephalexin RS in the CS Standard stock solution (mg/mL) = nominal concentration of cephalexin from the CU Sample stock solution (mg/mL) P = designated potency of USP Cephalexin RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Dilute with Medium as needed. Standard solution: 20 g/mL of USP Cephalexin RS at a known concentration in the Medium Spectrometric conditions Mode: UV Analytical wavelength: 262 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 80% (Q) of the labeled amount of C16H17N3O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 10.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cephalexin RS
Cephalexin for Oral Suspension

(Comment on this Monograph)id=m14300=Cephalexin for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cephalexin for Oral Suspension is a dry mixture of Cephalexin and one or more suitable buffers, colors, diluents, and flavors. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of C16H17N3O4S per mL when constituted as directed in the labeling. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 3 mg/mL of USP Cephalexin RS in water Sample solution: 3 mg/mL of cephalexin from Oral Suspension, constituted as directed in the labeling and filtered Chromatographic system Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1 in 15) of ninhydrin in acetone (60:40:1.5)
USP 32
= nominal concentration of cephalexin from the Sample solution (mg/mL) P = designated potency of USP Cephalexin RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 2.0% DELIVERABLE VOLUME 698: Meets the requirements PH 791: 3.06.0 PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 For solid packaged in single-unit containers: meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cephalexin RS CU

Standard stock solution: 1 mg/mL of USP Cephalexin RS in water Standard solution: Standard stock solution and Internal standard solution (2:3) Sample stock solution: Equivalent to 1 mg/mL of cephalexin from combined contents of powdered Tablets (NLT 20) in water. Sonicate, if necessary, to assure complete dissolution of the cephalexin. Filter, if necessary, to obtain a clear solution. Sample solution: Sample stock solution and Internal standard solution (2:3) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for 1hydroxybenzotriazole and cephalexin are about 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the internal standard and the analyte peaks, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O4S in the portion of Tablets taken: Result = (RU/RS) (CS/CU) P F 100 = peak response ratio of cephalexin to the 1hydroxybenzotriazole peak from the Sample solution = peak response ratio of cephalexin to the 1RS hydroxybenzotriazole peak from the Standard solution = concentration of USP Cephalexin RS in the CS Standard stock solution used to prepare the Standard solution (mg/mL) = nominal concentration of cephalexin taken to CU prepare the Sample solution (mg/mL) P = designated potency of USP Cephalexin RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% RU PERFORMANCE TESTS DISSOLUTION 711 Cephalexin Medium: Water; 900 mL Apparatus 1: Use 40-mesh cloth and 100 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: 20 g/mL of USP Cephalexin RS in the Medium Spectrometric conditions Mode: UV Analytical wavelength: 262 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 80% (Q) of the labeled amount of C16H17N3O4S is dissolved.
Cephalexin Tablets
(Comment on this Monograph)id=m14310=Cephalexin Tablets=Ca-Chl-Monos.pdf) DEFINITION Cephalexin Tablets are prepared from Cephalexin or Cephalexin Hydrochloride. They contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cephalexin (C16H17N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 3 mg/mL of USP Cephalexin RS in water Sample solution: 3 mg/mL of cephalexin from powdered Tablets in water and filter Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1 in 15) of ninhydrin in acetone (60:40:1.5) Analysis Samples: Standard solution and Sample solution Allow the solvent front to move the length of the plate in the Pre-developing solvent system. On this plate apply 10 L each of the Sample solution and Standard solution. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: 1.0 g of sodium 1-pentanesulfonate in a mixture of acetonitrile, methanol, triethylamine, and water (100:50:15:850). Adjust with phosphoric acid to a pH of 3.0 0.1. Internal standard solution: 0.3 mg/mL of 1hydroxybenzotriazole dissolved in methanol and diluted with Mobile phase (1:99)
190
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Internal standard solution: 0.3 mg/mL of 1hydroxybenzotriazole dissolved in methanol and diluted with Mobile phase (1:99) Standard stock solution: 1 mg/mL of USP Cephalexin RS in water Standard solution: Mix 10 mL of Standard stock solution and 15 mL of Internal Standard solution. Sample stock solution: Nominally equivalent to 1 mg/mL of cephalexin from combined contents of NLT 20 powdered Tablets for Oral Suspension in water. Pass a portion of the solution through a filter having a 1-m or finer porosity. Sample solution: Mix 10 mL of Sample stock solution with 15 mL of Internal Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 of low acidity Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for 1hydroxybenzotriazole and cephalexin are about 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5 between the internal standard and the analyte peaks, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H17N3O4S in each Tablet for Oral Suspension: Result = (RU/RS) (CS/CU) P F 100 = peak response ratio of cephalexin to the 1hydroxybenzotriazole peak from the Sample solution RS = peak response ratio of cephalexin to the 1hydroxybenzotriazole peak from the Standard solution CS = concentration of USP Cephalexin RS in the Sample stock solution (mg/mL) CU = nominal concentration of cephalexin in the Sample stock solution (mg/mL) P = designated potency of USP Cephalexin RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS DISINTEGRATION 701: Tablets for Oral Suspension disintegrate in 3 min, using water at 20 5. DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: Use 40-mesh cloth and 100 rpm Time: 30 min Spectrometric conditions Mode: UV Analytical wavelength: 262 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 80% (Q) of the labeled amount of C16H17N3O4S is dissolved. DISPERSION FINENESS: Place 2 Tablets for Oral Suspension in 100 mL of water, and stir until completely dispersed. A smooth dispersion is obtained that passes through a No. 25 sieve.
Cephalexin hydrochloride Medium, Sample solution, Spectrometric conditions, Standard solution, and Analysis: Proceed as directed under Cephalexin. Apparatus 1: Use 10-mesh cloth and 150 rpm Time: 45 min Tolerances: NLT 75% (Q) of the labeled amount of C16H17N3O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 9.0% where Tablets contain Cephalexin; NMT 8.0% where Tablets contain Cephalexin Hydrochloride ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The label states whether the Tablets contain Cephalexin or Cephalexin Hydrochloride. USP REFERENCE STANDARDS 11 USP Cephalexin RS
Cephalexin Tablets for Oral Suspension

(Comment on this Monograph)id=m323=Cephalexin Tablets for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cephalexin Tablets for Oral Suspension contain NLT 90.0% and NMT 110.0% of the labeled amount of cephalexin (C16H17N3O4S). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 3 mg/mL of USP Cephalexin RS in water Sample solution: 3 mg/mL of cephalexin from powdered Tablets for Oral Suspension in water and filter Chromatographic system (See Chromatography 621, Thin-Layer Chromatography. Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1 of 15) of ninhydrin in acetone (60:40:1.5) Analysis Samples: Standard solution and Sample solution Allow the solvent front to move the length of the plate in the Pre-developing solvent system, remove the plate from the chamber, and allow the solvent to evaporate. On this plate apply 10 L each of the Standard solution and Sample solution. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: 1.0 g of sodium 1-pentanesulfonate in the mixture of acetonitrile, methanol, triethylamine, and water (100:50:15:850), adjust with phosphoric acid to a pH of 3.0 0.1
USP 32
UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements NMT 9.0%
Official Monographs / Cephalothin 191

Tailing factor: NMT 1.8, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Samples: Standard solution and Sample solution [NOTEUse peak areas where peak responses are indicated.] Calculate the quantity of C16H16N2O6S2, in g, in each mg of Cephalothin Sodium taken: Result = (rU/rS) (CS/CU) P
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cephalexin RS
Cephalothin Sodium
(Comment on this Monograph)id=m14455=Cephalothin Sodium=Ca-Chl-Monos.pdf)
= peak response of cephalothin from the Sample solution = peak response of cephalothin from the Standard rS solution = concentration of USP Cephalothin Sodium RS in CS the Standard solution (mg/mL) = concentration of Cephalothin Sodium taken to CU prepare the Sample solution (mg/mL) P = content of cephalothin in USP Cephalothin RS (g/mg) Acceptance criteria: NLT 850 g/mg IMPURITIES Organic Impurities PROCEDURE Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Standard solution: 0.01 mg/mL of USP Cephalothin Sodium RS Sample solution: Use the Sample solution prepared as directed in the Assay. Analysis Samples: Standard solution and Sample solution Proceed as directed for Analysis in the Assay, except to continue the chromatography of the Sample solution for at least 4 times the retention time of the main cephalothin peak. Acceptance criteria: The area of any peak from the Sample solution, except the main peak, is NMT the area of the main peak in the Standard solution (1.0%), and the sum of the areas of any such peaks is NMT 3 times the area of the main peak from the Standard solution (3.0%). [NOTEAny peak from the Sample solution with an area less than one-tenth that of the main peak from the Standard solution is to be disregarded.] SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +124 to +134 Sample solution: A known amount of specimen, equivalent to 50 mg/mL of cephalothin, in water CRYSTALLINITY 695: Meets the requirements PH 791: 4.57.0, in a solution containing 250 mg/mL LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 1.5% of its weight. STERILITY TESTS 71: Meets the requirements, when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cephalothin Sodium must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements. It contains NMT 0.13 USP Endotoxin Unit/mg of cephalothin. OTHER REQUIREMENTS: Where the label states that Cephalothin Sodium is sterile, it meets the requirements.
rU
418.42 C16H15N2NaO6S2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3[(acetyloxy)methyl]-8-oxo-7-[(2-thienylacetyl)amino]-, monosodium salt, (6R-trans)-; Monosodium (6R,7R)-3-(hydroxymethyl)-8-oxo-7-[2-(2-thienyl)acetamido]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate acetate (ester) [58-71-9]. DEFINITION Cephalothin Sodium contains the equivalent of NLT 850 g of C16H16N2O6S2 per mg, calculated on the dried basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: 25 g/mL in water B. IDENTIFICATION TESTSGENERAL, Sodium 191: requirements
Meets the
ASSAY PROCEDURE Mobile phase: Dissolve 17 g of sodium acetate in 790 mL of water, add 0.6 mL of glacial acetic acid, and if necessary, adjust with 0.1 N sodium hydroxide or glacial acetic acid to a pH of 5.9 0.1. Add 150 mL of acetonitrile and 70 mL of alcohol. Standard solution: 1 mg/mL of USP Cephalothin Sodium RS in Mobile phase System suitability solution: Heat a 5-mL portion of the Standard solution in a water bath at 90 for 10 min. Cool the solution, and immediately inject a portion of it into the chromatograph as directed under Chromatographic system. Sample solution: 1 mg/mL of Cephalothin Sodium in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5 m packing L1 Temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 9.0 between the two principal peaks, System suitability solution
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P
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= assigned potency of USP Cephalothin Sodium RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%115.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: NMT 0.13 USP Endotoxin Unit/mg of cephalothin PH 791: 6.08.5 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Injections as described under Injections 1. Maintain in the frozen state. LABELING: It meets the requirements for Labeling under Injections 1. The label states that it is to be thawed just prior to use, describes conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Cephalothin Sodium RS USP Endotoxin RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cephalothin Sodium RS USP Endotoxin RS
Cephalothin Injection
(Comment on this Monograph)id=m14457=Cephalothin Injection=Ca-Chl-Monos.pdf) DEFINITION Cephalothin Injection contains an amount of Cephalothin Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of cephalothin (C16H16N2O6S2). IDENTIFICATION ULTRAVIOLET ABSORPTION 197U Sample solution: 25 g/mL in water ASSAY PROCEDURE Mobile phase: 17 g of sodium acetate in 790 mL of water, add 0.6 mL of glacial acetic acid, and, if necessary, adjust with 0.1 N sodium hydroxide or glacial acetic acid to a pH of 5.9 0.1. Add 150 mL of acetonitrile and 70 mL of alcohol. Standard solution: 1 mg/mL of USP Cephalothin Sodium RS in Mobile phase System suitability solution: Heat a 5-mL portion of the Standard solution in a water bath at 90 for 10 min. Cool the solution, and immediately inject a portion of it into the chromatograph as directed under Chromatographic system. Sample solution: 1 mg/mL of cephalothin sodium in the Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 9.0 between the two principal peaks, System suitability solution Tailing factor: NMT 1.8, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H16N2O6S2 in each mL of the Injection: Result = (rU/rS) (CS/CU) P F 100 rU rS CS CU = peak response of cephalothin from the Sample solution = peak response of cephalothin from the Standard solution = concentration of USP Cephalothin Sodium RS in the Standard solution (mg/mL) = nominal concentration of cephalothin sodium in the Sample solution (mg/mL)
Cephalothin for Injection

(Comment on this Monograph)id=m14461=Cephalothin for Injection=Ca-Chl-Monos.pdf) DEFINITION Cephalothin for Injection contains an amount of Cephalothin Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of cephalothin (C16H16N2O6S2). It may contain Sodium Bicarbonate. IDENTIFICATION ULTRAVIOLET ABSORPTION 197U Sample solution: 25 g/mL in water ASSAY PROCEDURE Mobile phase: 17 g of sodium acetate in 790 mL of water, add 0.6 mL of glacial acetic acid, and if necessary adjust with 0.1 N sodium hydroxide or glacial acetic acid to a pH of 5.9 0.1. Add 150 mL of acetonitrile and 70 mL of alcohol Standard solution: 1 mg/mL of USP Cephalothin Sodium RS in Mobile phase System suitability solution: Heat a 5-mL portion of the Standard solution in a water bath at 90 for 10 min. Cool the solution, and immediately inject a portion of it into the chromatograph as directed under Chromatographic system. Sample solution A (where it is represented as being in a single-dose container): Constitute Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute quantitatively with Mobile phase to obtain a solution having a concentration of about 1 mg of cephalothin/mL. Sample solution B (where the label states the quantity of cephalothin in a given volume of constituted solution): Constitute 1 container of Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute an accurately measured portion of the constituted solution quantitatively with the Mobile phase to
USP 32
obtain a solution having a concentration of about 1 mg of cephalothin/mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L1 Temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 9.0 between the two principal peaks, System suitability solution Tailing factor: NMT 1.8, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Samples: Standard solution and Sample solution A or Sample solution B Calculate the percentage of C16H16N2O6S2 in the container, and in the portion of constituted solution: Result = (rU/rS) (CS/CU) P F 100 = cephalothin peak response from the Sample solution = cephalothin peak response from the Standard rS solution = concentration of USP Cephalothin Sodium RS in CS the Standard solution (mg/mL) = nominal concentration of cephalothin in Sample CU solution A or Sample solution B, based on the labeled quantity in the container or in the portion of constituted solution or suspension taken, and the extent of dilution P = assigned potency of USP Cephalothin Sodium RS (g/mg) F = unit conversion factor, 0.001 mg/g [NOTEWhere the test for Uniformity of dosage units has been performed using the Analysis for content uniformity, use the average of these determinations as the Assay value.] Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Analysis for content uniformity: Perform the Assay on individual containers using Sample solution A or Sample solution B, or both, as appropriate. SPECIFIC TESTS CONTENT OF SODIUM BICARBONATE (if present) Sample: 1 g Analysis: Dissolve in 50 mL of water. Add methyl orange TS, and titrate with 0.1 N sulfuric acid VS. Each mL of 0.1 N sulfuric acid is equivalent to 8.401 mg of NaHCO3. Calculate the percentage of sodium bicarbonate, and use the value obtained to calculate the Specific rotation on the dried and sodium bicarbonate-free basis. INJECTIONS, Constituted Solutions 1: At the time of use, it meets the requirements. OPTICAL ROTATION, Specific Rotation 781S: +124 to +134, calculated on the dried and sodium bicarbonate-free basis Sample solution: Equivalent to 50 mg/mL, in water BACTERIAL ENDOTOXINS TEST 85: NMT 0.13 USP Endotoxin Unit/mg of cephalothin STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. rU
Official Monographs / Cephapirin 193

PH 791: 6.08.5, in the solution constituted as directed in the labeling PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 1.5% of its weight. INJECTIONS, Labeling 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Cephalothin Sodium RS USP Endotoxin RS
Cephapirin Benzathine
(Comment on this Monograph)id=m14490=Cephapirin Benzathine=Ca-Chl-Monos.pdf)
1087.30 (C17H17N3O6S2)2 C16H20N2 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 3-[(acetyloxy)methyl]-8-oxo-7-[[(4-pyridinylthio)acetyl]amino]-, (6Rtrans)-, compd. with N,N-bis(phenylmethyl)-1,2ethanediamine (2:1); (6R,7R)-3-(Hydroxymethyl)-8-oxo-7-[2-(4pyridylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2carboxylic acid compound with N,N-dibenzylethylenediamine (2:1) [97468-37-6]. DEFINITION Cephapirin Benzathine contains the equivalent of NLT 715 g and NMT 820 g of cephapirin (C17H17N3O6S2) per mg. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Solution A: 26.2 mL of acetic acid and 99.12 g of potassium acetate to a 4-L volumetric flask. Add 2000 mL of water, and mix to dissolve. Dilute with water to volume, and pass through a 0.45-m nylon filter. Solution B: Acetonitrile Mobile phase: See the gradient table below.
Time (min) 0 6 10 12 21 Solution A (%) 91.5 91.5 80.0 80.0 91.5 Solution B (%) 8.5 8.5 20.0 20.0 8.5
Solution C: Acetic acid and water (2:3) Diluent: 205 mg/mL of potassium acetate in water. Adjust with acetic acid to a pH of 7.5 to 8.2. Pass through a 0.45m nylon filter.
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ether extract to the water-washed combined ether extracts. Evaporate this combined ether solution to a volume of 5 mL, add 2 mL of dehydrated alcohol, and evaporate to dryness. Dissolve the residue in 50 mL of glacial acetic acid, add 1 mL of p-naphtholbenzein TS, and titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 12.02 mg of benzathine (C16H20N2). Acceptance criteria: Between 20.0% and 24.0%, calculated on the anhydrous basis SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 4.07.0, in a suspension (1 in 10) WATER DETERMINATION, Method I 921: NMT 5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Cephapirin Benzathine RS USP Cephapirin Sodium RS
Solution D: Acetic acid and water (1:9) System suitability solution: 2.0 mg/mL of USP Cephapirin Sodium RS in Solution D. Heat the solution at 50 for 1218 h. Standard solution: In duplicate, 50 mg of USP Cephapirin Sodium RS, and transfer into a 25-mL volumetric flask. Add 2.5 mL of Solution C and 15.0 mL of Diluent, and agitate to dissolve. Add 7.0 mL of acetonitrile, and mix well. Allow the solution to return to room temperature, and dilute with water to volume. Sample solution: In duplicate, 60 mg of Cephapirin Benzathine, and transfer into a 25-mL volumetric flask. Add 2.5 mL of Solution C and 15.0 mL of Diluent, and mix to dissolve. Add 7.0 mL of acetonitrile, and mix. Allow the flask to return to room temperature, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 260 nm Column: 3.9-mm 15-cm; 4-m packing L1 Guard column: 3.2-mm 15-mm; 7-m packing L1 Temperature: 40 Flow rate: 2 mL/min Injection size: 2 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Using the results from the System suitability solution, calculate the percentage of the height of the valley taken: Result = (RV/RU) 100 = height of the valley between cephapirin and any impurity = impurity peak height RU The percentage of the height of the valley is NMT 25% for the impurity peaks adjacent to the cephapirin peak. [NOTEThe System suitability solution is acceptable as long as the cephapirin peak is larger than the two peaks on either side of the cephapirin peak.] Relative standard deviation: NMT 3.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity of C17H17N3O6S2 in each mg of Cephapirin Benzathine taken: RV Result = (rU/rS) (WS/WU) (VU/VS) P = average peak areas of the cephapirin peaks from the Sample solution = average peak areas of the cephapirin peaks from rS the Standard solution = quantity of USP Cephapirin Sodium RS used to WS prepare the Standard solution (mg) = quantity of Cephapirin Benzathine used to WU prepare the Sample solution (mg) = final volume of the Sample solution (mL) VU = final volume of the Standard solution (mL) VS P = assigned potency of USP Cephapirin Sodium RS (g/mg) Acceptance criteria: 715820 g/mg rU OTHER COMPONENTS BENZATHINE CONTENT Sample: 1 g of Cephapirin Benzathine Analysis: Add 30 mL of a saturated solution of sodium chloride and 10 mL of 5 N sodium hydroxide, and extract with four 50-mL portions of ether. Wash the combined ether extracts with three 10-mL portions of water. Extract the combined water washings with 25 mL of ether, and add the
Cephapirin Benzathine Intramammary Infusion

(Comment on this Monograph)id=m14495=Cephapirin Benzathine Intramammary Infusion=Ca-Chl-Monos.pdf) DEFINITION Cephapirin Benzathine Intramammary Infusion is a suspension of Cephapirin Benzathine in a suitable vegetable oil vehicle. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled quantity of cephapirin (C17H17N3O6S2). It contains a suitable dispersing agent. IDENTIFICATION INFRARED ABSORPTION 197K Sample solution: Transfer the contents of 1 syringe of Intramammary Infusion to a 50-mL centrifuge tube, add 25 mL of toluene, mix for 1 min, and centrifuge. Remove and discard the toluene layer without disturbing the residue in the centrifuge tube. Wash the residue with two 25-mL portions of toluene. Dry the residue in a vacuum at 60, and use the dried residue as the sample. Mix the dried residue with 9 parts of potassium bromide, and record the IR spectrum, using the diffuse reflectance technique. Acceptance criteria: The IR absorption spectrum so obtained corresponds to that of a similar dispersion of USP Cephapirin Benzathine RS in potassium bromide. ASSAY PROCEDURE Solution A: Transfer 26.2 mL of acetic acid and 99.12 g of potassium acetate to a 4-L volumetric flask. Add 2000 mL of water, and mix to dissolve. Dilute with water to volume, and pass through a 0.45-m nylon filter. Solution B: Acetonitrile Mobile Phase: See the gradient table below.
Time (min) 0 6 10 Solution A (%) 91.5 91.5 80.0 Solution B (%) 8.5 8.5 20.0
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Time (min) 12 21 Solution A (%) 80.0 91.5 Solution B (%) 20.0 8.5

= quantity of cephapirin benzathine used to prepare the Sample solution (mg) = entire volume in one syringe (volume of VU Intramammary Infusion in mL) VS = final volume of the Standard solution (mL) P = assigned potencyof USP Cephapirin Sodium RS (g cephapirin/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1.0%, 10 mL of Intramammary Infusion being tested ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed unit-dose disposable syringes at controlled room temperature. LABELING: Label Intramammary Infusion to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Cephapirin Benzathine RS USP Cephapirin Sodium RS WU
Solution C: Acetic acid and water (2:3) Diluent: 205 mg/mL of potassium acetate in water. Adjust with acetic acid to a pH of 7.58.2. Pass through a 0.45-m nylon filter. Solution D: Acetic acid and water (1:9) System suitability solution: 2.0 mg/mL of USP Cephapirin Sodium RS in Solution D. Heat the solution at 50 for 1218 h. Standard solution: In duplicate, weigh 50 mg of USP Cephapirin Sodium RS, and transfer into a 25-mL volumetric flask. Add 2.5 mL of Solution C and 15.0 mL of Diluent, and agitate to dissolve. Add 7.0 mL of acetonitrile, and mix well. Allow the solution to return to room temperature, and dilute with water to volume. Sample solution: Express the entire contents of a syringe of the Intramammary Infusion into a centrifuge tube. For each mL of Intramammary Infusion, add 1.0 mL of n-heptane and 1.5 mL of Solution C, cap, and vortex at high speed for 5 min. Centrifuge for 5 min at a sufficient speed to break the emulsion. Remove the aqueous layer, and pass through a 0.45-m nylon filter, discarding the first 0.5 mL. Transfer 2.5 mL of the filtered aqueous phase into a 25-mL volumetric flask that contains a solution composed of 15.0 mL of Diluent and 7.0 mL of acetonitrile. Add water to volume, and mix well to obtain a single phase. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 260 nm Column: 3.9-mm 15-cm; 4-m packing L1 Guard column: 3.2-mm 15-mm; 7-m packing L1 Temperature: 40 Flow rate: 2 mL/min Injection size: 2 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Relative standard deviation: NMT 3.0%, Standard solution Using the results from the System suitability solution, calculate the percentage of the height of the valley taken: Result = (rV/ri) 100 = height of the valley between cephapirin and any impurity = impurity peak height ri Acceptance criteria: The percentage of the height of the valley is NMT 25% for the impurity peaks adjacent to the cephapirin peak. [NOTEThe System suitability solution is acceptable as long as the cephapirin peak is larger than the two peaks on either side of the cephapirin peak.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H17N3O6S2 in each syringe of Intramammary Infusion taken: rV (rU/rS) (WS/WU) (VU/VS) P F 100 rU rS WS = peak area of the cephapirin peak from the Sample solution = average peak area of the cephapirin peak from the Standard solution = quantity of USP Cephapirin Sodium RS used to prepare the Standard solution (mg)
Cephapirin Sodium
(Comment on this Monograph)id=m14510=Cephapirin Sodium=Ca-Chl-Monos.pdf)
C17H16N3NaO6S2 445.45 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid, 3[(acetyloxy)methyl]-8-oxo-7-[[(4-pyridinylthio)acetyl]amino]-, monosodium salt, (6R-trans)-; Monosodium (6R,7R)-3-(hydroxymethyl)8-oxo-7-[2-(4-pyridylthio)acetamido]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate acetate (ester) [24356-60-3]. DEFINITION Cephapirin Sodium has a potency equivalent to NLT 855 g and NMT 1000 g of cephapirin (C17H17N3O6S2) per mg. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sodium 191: requirements
Meets the
ASSAY PROCEDURE Solution A: 26.2 mL of acetic acid and about 99.12 g of potassium acetate to a 4-L volumetric flask. Add 2000 mL of water, and mix to dissolve. Dilute with water to volume, and pass through a 0.45-m nylon filter. Solution B: Acetonitrile Mobile phase: See the gradient table below.
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WATER DETERMINATION, Method I 921: NMT 2.0% STERILITY TESTS 71: Where the label states that Cephapirin Sodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cephapirin Sodium is sterile or it must be subjected to further processing during the preparation of injectable dosage forms, it has NMT 0.17 USP Endotoxin Unit/mg of cephapirin. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cephapirin Sodium RS USP Endotoxin RS
Solution C: Acetic acid and water (2:3) Diluent: 205 mg/mL of potassium acetate in water. Adjust with acetic acid to a pH of 7.58.2. Pass through a 0.45-m nylon filter. Solution D: Acetic acid and water (1:9) System suitability solution: 2.0 mg/mL of USP Cephapirin Sodium RS in Solution D. Heat the solution at 50 for 1218 h. Standard solution: In duplicate, 50 mg of USP Cephapirin Sodium RS, and transfer into a 25-mL volumetric flask. Add 2.5 mL of Solution C and 15.0 mL of Diluent, and agitate to dissolve. Add 7.0 mL of acetonitrile, and mix well. Allow the solution to return to room temperature, and dilute with water to volume. Sample solution: In duplicate, 50 mg of Cephapirin Sodium, and transfer into a 25-mL volumetric flask. Add 2.5 mL of Solution C and 15.0 mL of Diluent, and mix to dissolve. Add 7.0 mL of acetonitrile. Allow the flask to return to room temperature, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 260 nm Column: 3.9-mm 15-cm; 4-m packing L1 Guard column: 3.2-mm 15-mm; 7-m packing L1 Temperature: 40 Flow rate: 2 mL/min Injection size: 2 L System suitability Samples: System suitability solution and Standard solution Suitability requirements: Use the results from the System suitability solution. Calculate the percentage of the height of the valley taken: Result = (rV/rU) 100 = height of the valley between cephapirin and any impurity rU = impurity peak height The percentage of the height of the valley is NMT 25% for the impurity peaks adjacent to the cephapirin peak. [NOTEThe System suitability solution is acceptable as long as the cephapirin peak is larger than the two peaks on either side of the cephapirin peak.] Relative standard deviation: NMT 3.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C17H17N3O6S2 in each mg of Cephapirin Sodium: rV Result = (rU/rS) (WS/WU) (VU/VS) P = average peak areas of the cephapirin peaks from the Sample solution = average peak areas of the cephapirin peaks from rS the Standard solution = quantity of USP Cephapirin Sodium RS used to WS prepare the Standard solution (mg) = quantity of Cephapirin Sodium used to prepare WU the Sample solution (mg) = final volume of the Sample solution (mL) VU = final volume of the Standard solution (mL) VS P = assigned potency of cephapirin/mg of USP Cephapirin Sodium RS (g) Acceptance criteria: 8551000 g/mg rU SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 6.58.5, in a solution containing 10 mg/mL of cephapirin
Cephapirin for Injection

(Comment on this Monograph)id=m14514=Cephapirin for Injection=Ca-Chl-Monos.pdf) DEFINITION Cephapirin for Injection contains an amount of Cephapirin Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of cephapirin. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sodium 191: the requirements.
It meets
ASSAY PROCEDURE Mobile phase: Dimethylformamide, glacial acetic acid, 11.7 N potassium hydroxide, and water (160:4:2:1834) [NOTE Increase the proportion of dimethylformamide to decrease the retention time of cephapirin.] System suitability solution: 1 mg/mL of Cephapirin Sodium in pH 2.0 hydrochloric acid buffer (see Reagents, Indicators, and SolutionsBuffer Solutions). Place 10 mL of this solution in a test tube, and heat at 95 for 10 min, accurately timed. Promptly cool the tube in an ice-water bath. Dilute 5 mL of the cooled solution with Mobile phase to obtain 50 mL of System suitability solution. Standard solution: 0.2 mg/mL of USP Cephapirin Sodium RS in Mobile phase Sample solution A (where it is packaged for dispensing and is represented as being in a single-dose container): Constitute Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with Mobile phase to obtain a solution containing the nominal equivalent of 0.2 mg of cephapirin/mL. Sample solution B (where the label states the quantity of cephapirin in a given volume of constituted solution): Constitute Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a volume of the constituted solution with Mobile phase to obtain a solution containing the nominal equivalent of 0.2 mg of cephapirin/mL. [NOTEUse the Standard solution and the Sample solutions within 1 h.] Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cephapirin lactone and cephapirin are about 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 0.9 between the cephapirin peak and the peak having a retention time of 0.9 relative to that of cephapirin, System suitability solution Column efficiency: NLT 1200 theoretical plates, Standard solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of label claim A or Sample solution B, in mg, of C17H17N3O6S2)withdrawn from the container, or in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) P F 100 = cephapirin peak response from the Sample solution = cephapirin peak response from the Standard rS solution = concentration of USP Cephapirin Sodium RS in CS the Standard solution (mg/mL) = nominal concentration of cephapirin in Sample CU solution A or Sample solution B (mg/mL) P = designated potency of USP Cephapirin Sodium RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements rU
Cephapirin Sodium Intramammary Infusion

(Comment on this Monograph)id=m14540=Cephapirin Sodium Intramammary Infusion=Ca-Chl-Monos.pdf) DEFINITION Cephapirin Sodium Intramammary Infusion is a suspension of Cephapirin Sodium in a suitable vegetable oil vehicle. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled quantity of cephapirin (C17H17N3O6S2). It contains a suitable dispersing agent. IDENTIFICATION INFRARED ABSORPTION 197K Sample solution: Transfer the contents of 1 syringe of Intramammary Infusion to a 50-mL centrifuge tube, add 25 mL of toluene, mix for 1 min, and centrifuge. Remove and discard the toluene layer without disturbing the residue in the centrifuge tube. Wash the residue with two 25-mL portions of toluene. Dry the residue in a vacuum at 60, and use the dried residue as the sample. Mix the dried residue with 9 parts of potassium bromide, and record the infrared spectrum, using the diffuse reflectance technique. ASSAY PROCEDURE Solution A: 26.2 mL of acetic acid and 99.12 g of potassium acetate to a 4-L volumetric flask. Add 2000 mL of water, and mix to dissolve. Dilute with water to volume, and pass through a 0.45-m nylon filter. Solution B: Acetonitrile Mobile phase: See gradient table below.
SPECIFIC TESTS INJECTIONS, Constituted Solutions 1: At the time of use, it meets the requirements. BACTERIAL ENDOTOXINS TEST 85: NMT 0.17 USP Endotoxin Unit/mg of cephapirin STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections CRYSTALLINITY 695: Meets the requirements PH 791: 6.58.5, in a solution containing 10 mg/mL of cephapirin WATER DETERMINATION, Method I 921: NMT 2.0% INJECTIONS, Labeling 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in Containers for Sterile Solids under Injections 1. USP REFERENCE STANDARDS 11 USP Cephapirin Sodium RS USP Endotoxin RS
Solution C: Acetic acid and water (2:3) Diluent: 205 mg/mL of potassium acetate in water. Adjust with acetic acid to a pH of 7.5 to 8.2. Pass through a 0.45m nylon filter. Solution D: Acetic acid and water (1:9) System suitability solution: 2.0 mg/mL of USP Cephapirin Sodium RS in Solution D. Heat the solution at 50 for 1218 h. Standard solution: In duplicate, 50 mg of USP Cephapirin Sodium RS, and transfer into a 25-mL volumetric flask. Add 2.5 mL of Solution C and 15.0 mL of Diluent, and agitate to dissolve. Add 7.0 mL of acetonitrile, and mix well. Allow the solution to return to room temperature, and dilute with water to volume. Sample solution: Express the entire contents of a syringe of the Intramammary Infusion into a centrifuge tube. For each mL of Intramammary Infusion, add 1.0 mL of n-heptane and 1.0 mL of Solution C, cap, and mix on a vortex mixer at high speed for 5 min. Centrifuge for 5 min at a speed sufficient to break the emulsion. Remove the aqueous layer,
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and pass through a 0.45-m nylon filter, discarding the first 0.5 mL. Transfer 2.5 mL of the filtered aqueous phase into a 25-mL volumetric flask that contains a solution composed of 15.0 mL of Diluent and 7.0 mL of acetonitrile. Add water to volume, and mix well to obtain a single phase. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 260 nm Column: 3.9-mm 15-cm; 4-m packing L1 Guard column: 3.2-mm 15-mm; 7-m packing L1 Temperature: 40 Flow rate: 2 mL/min Injection size: 2 L System suitability Samples: System suitability solution and Standard solution Suitability requirements: Using the results from the System suitability solution, calculate the percentage of the height of the valley taken: Result = (rV/rI) 100 = height of the valley between cephapirin and any impurity = impurity peak height rI The percentage of the height of the valley is NMT 25% for the impurity peaks adjacent to the cephapirin peak. [NOTEThe System suitability solution is acceptable as long as the cephapirin peak is larger than the two peaks on either side of the cephapirin peak.] Relative standard deviation: NMT 3.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H17N3O6S2 in each syringe of Intramammary Infusion: rV Result = (rU/rS) (CS/CU) P F 100 = peak areas of the cephapirin peaks from the Sample solution = average peak areas of the cephapirin peaks from rS the Standard solution = concentration of USP Cephapirin Sodium RS of CS the Sample solution (mg/mL) = nominal concentration of cephapirin sodium CU from the Standard solution (mg/mL) P = assigned potency of USP Cephapirin Sodium RS in g cephapirin/mg F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% rU SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1.0%, 10 mL of Intramammary Infusion being tested ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, unit-dose disposable syringes at controlled room temperature. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Cephapirin Sodium RS
Cephradine
(Comment on this Monograph)id=m14570=Cephradine=CaChl-Monos.pdf)
349.41 C16H19N3O4S 5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7[(amino-1,4-cyclohexadien-1-ylacetyl)amino]-3-methyl-8-oxo-, [6R-[6,7 (R*)]]-; (6R,7R)-7-[(R)-2-Amino-2-(1,4-cyclohexadien-1-yl)acetamido]-3methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid [38821-53-3]. Monohydrate 367.43 [31828-50-9 (non-stoichiometric hydrate)]. Dihydrate 385.44 [58456-86-3]. DEFINITION Cephradine has a potency of NLT 900 g and NMT 1050 g of total cephalosporins per mg, calculated as the sum of cephradine (C16H19N3O4S) and cephalexin (C16H17N3O4S), calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Mobile phase: Methanol, 0.5 M sodium acetate, 0.7 N acetic acid, and water (200:15:3:782). Pass the solution through a filter of 1 m or finer porosity. System suitability solution: 0.5 mg/mL of USP Cephradine RS and 0.5 mg/mL of USP Cephalexin RS in Mobile phase Standard solution: 0.5 mg/mL of USP Cephradine RS in Mobile phase Sample solution: 0.5 mg/mL of Cephradine in Mobile phase [NOTESonicate prior to dilution to volume.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cephalexin and cephradine are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the cephalexin peak and the cephradine peak, System suitability solution. Relative standard deviation: NMT 2.0%, Standard solution
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of total cephalosporins (sum of cephradine and cephalexin) in each mg of the Cephradine taken: Result = (rU/rS) (CS/CU) P = sum of the cephradine and cephalexin peak response from the Sample solution = sum of the cephradine and cephalexin peak rS response from the Standard solution = concentration of USP Cephradine RS from the CS Standard solution (mg/mL) = concentration of Cephradine taken to prepare CU the Sample solution (mg/mL) P = potency of USP Cephradine RS (g/mg) Acceptance criteria: 9001050 g IMPURITIES Organic Impurity PROCEDURE: LIMIT OF CEPHALEXIN: Using the chromatogram of the Sample solution in the Assay, calculate the percentage of C16H17N3O4S in the portion of Cephradine taken: Result = (rux/rU) 100 = peak response of the cephalexin from the Sample solution = sum of the cephalexin and cephradine peak rU responses in the chromatogram from the Sample solution Acceptance criteria: NMT 5.0% rux SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 3.56.0, in a solution containing 10 mg/mL WATER DETERMINATION, Method I 921: NMT 6.0%, except that if it is the dihydrate form, the limit is between 8.5%10.5% STERILITY TESTS 71: Where the label states that Cephradine is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cephradine is sterile or it must be subjected to further processing during the preparation of injectable dosage forms, it has NMT 0.20 USP Endotoxin Units/mg of cephradine. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is the dihydrate form, the label so indicates. Where the quantity of cephradine is indicated in the labeling of any preparation containing Cephradine, this shall be understood to be in terms of anhydrous cephradine (C16H19N3O4S). Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cephalexin RS USP Cephradine RS USP Endotoxin RS rU
Official Monographs / Cephradine 199
Cephradine Capsules
(Comment on this Monograph)id=m14580=Cephradine Capsules=Ca-Chl-Monos.pdf) DEFINITION Cephradine Capsules contain NLT 90.0% and NMT 120.0% of the labeled amount of cephradine, calculated as the sum of cephradine (C16H19N3O4S) and cephalexin (C16H17N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 3 mg/mL of USP Cephradine RS in water Sample solution: 3 mg/mL of cephradine from Capsule content in water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1-in-15) of ninhydrin in acetone (60:40:1.5) Analysis Samples: Standard solution and Sample solution Place the thin-layer chromatographic plate in a chamber containing the Pre-developing solvent system to a depth of 1 cm, allow the solvent front to move the length of the plate, remove the plate from the chamber, and allow the solvent to evaporate. Apply the Samples. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved threefourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, 0.5 M sodium acetate, 0.7 N acetic acid, and water (200:15:3:782). Pass the solution through a filter of 1-m or finer porosity. System suitability solution: 0.5 mg/mL of USP Cephradine RS and 0.5 mg/mL of USP Cephalexin RS in Mobile phase Standard solution: 0.5 mg/mL of USP Cephradine RS in Mobile phase Sample solution: Transfer the contents of NLT 20 Capsules to a tared container and determine the average weight/Capsule, and mix combined contents. Transfer a portion of the powder, nominally equivalent to 125 mg to a 250-mL volumetric flask. Add 50 mL of Mobile phase, sonicate for 15 min, and shake for 10 min. Dilute with Mobile phase to volume and pass a portion through a filter with a porosity of 0.5 m or finer. Chromatographic system (See Chromatography 621, System Suitability.)
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Cephradine / Official Monographs
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Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cephalexin and cephradine are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the cephalexin peak and the cephradine peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of cephradine (sum of cephradine and cephalexin) in the portion of Capsules taken: Result = (rU/rS) (CS/CU) P F 100 = sums of the cephradine and cephalexin peak response from the Sample solution = sums of the cephradine and cephalexin peak rS response from the Standard solution = concentration of USP Cephradine RS in the CS Standard solution (mg/mL) = nominal concentration of Cephradine taken to CU prepare the Sample solution (mg/mL) P = designated potency of USP Cephradine RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120% rU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.12 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 45 min Sample solution: Sample per Dissolution 711. Dilute with Medium as needed. Standard solution: USP Cephradine RS in Medium Spectrometric conditions Mode: UV Analytical wavelength: 255 nm Analysis Samples: Sample solution and Standard solution Tolerances: NLT 75% (Q) of the labeled amount of C16H19N3O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS LOSS ON DRYING 731: Dry 100 mg of the mixed contents of 4 Capsules in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: the Capsule contents lose NMT 7.0% of their weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: The quantity of cephradine stated in the labeling is in terms of anhydrous cephradine (C16H19N3O4S). USP REFERENCE STANDARDS 11 USP Cephalexin RS USP Cephradine RS
Cephradine for Injection

(Comment on this Monograph)id=m14590=Cephradine for Injection=Ca-Chl-Monos.pdf) DEFINITION Cephradine for Injection contains NLT 90.0% and NMT 115.0% of the labeled amount of cephradine, calculated as the sum of cephradine (C16H19N3O4S) and cephalexin (C16H17 N3O4S). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 3 mg/mL of USP Cephradine RS in water Sample solution: 3 mg/mL of cephradine from 1 container of Injection in water Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: Of 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1 in 15) of ninhydrin in acetone (60:40:1.5) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Analysis Samples: Standard solution and Sample solution Place a chromatographic plate in a chamber containing the Pre-developing solvent system to a depth of about 1 cm, allow the solvent front to move the length of the plate, remove the plate from the chamber, and allow the solvent to evaporate. Apply the Samples. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved about threefourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, 0.5 M sodium acetate, 0.7 N acetic acid, and water (200:15:3:782). Pass the solution through a filter of 1m or finer porosity. System suitability solution: 0.5 mg/mL of USP Cephradine RS and 0.5 mg/mL of USP Cephalexin RS in Mobile phase Standard solution: 0.5 mg/mL of USP Cephradine RS in Mobile phase Sample solution A (where it is represented as being in a single-dose container): Constitute Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with Mobile phase to obtain a solution containing nominally 0.5 mg of cephradine per mL. Sample solution B (where the label states the quantity of cephradine in a given volume of constituted solution): Constitute Injection in a volume of water corresponding to the volume of solvent specified in the labeling. Dilute a volume of the constituted solution with Mobile phase to
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obtain a solution containing nominally 0.5 mg of cephradine per mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cephalexin and cephradine are 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the cephalexin peak and the cephradine peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution A or Sample solution B Calculate the percentage of cephradine (sum of cephradine and cephalexin) withdrawn from the container, or in the portion of constituted solution: Result = (rU/rS) (CS/CU) P F 100 = sums of the cephradine and cephalexin peak response from Sample solution A or Sample solution B = sums of the cephradine and cephalexin peak rS response from the Standard solution = concentration of USP Cephradine RS in the CS Standard solution (mg/mL) = nominal concentration of cephradine from CU Sample solution A or Sample solution B (mg/mL) P = designated potency of USP Cephradine RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%115.0% rU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Official Monographs / Cephradine 201

USP Endotoxin RS
Cephradine for Oral Suspension

(Comment on this Monograph)id=m14610=Cephradine for Oral Suspension=Ca-Chl-Monos.pdf) DEFINITION Cephradine for Oral Suspension is a dry mixture of Cephradine and one or more suitable buffers, colors, diluents, and flavors. It contains NLT 90.0% and NMT 125.0% of the labeled amount of cephradine, calculated as the sum of cephradine (C16H19N3O4S) and cephalexin (C16H17N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 3 mg/mL of USP Cephradine RS in water Sample solution: Constitute 1 container of Oral Suspension as directed in the labeling. Mix a portion of the resulting suspension with water to obtain a concentration of about 3 mg/mL of cephradine and filter. Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: Of 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1 in 15) of ninhydrin in acetone (60:40:1.5) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Analysis Samples: Standard solution and Sample solution Place a chromatographic plate in a chamber containing the Pre-developing solvent system to a depth of 1 cm, allow the solvent front to move the length of the plate, remove the plate from the chamber, and allow the solvent to evaporate. Apply the Samples. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, 0.5 M sodium acetate, 0.7 N acetic acid, and water (200:15:3:782). Pass the solution through a filter of 1m or finer porosity System suitability solution: 0.5 mg/mL of USP Cephradine RS and 0.5 mg/mL of USP Cephalexin RS in Mobile phase Standard solution: 0.5 mg/mL of USP Cephradine RS in Mobile phase Sample solution: Constitute Oral Suspension as directed in the labeling. Dilute an accurately measured volume of the suspension so obtained, freshly mixed and free from air bubbles, in Mobile phase to obtain a solution containing about 0.5 mg of cephradine per mL. Pass a portion of this mixture through a filter having a porosity of 0.5-m or finer, discarding the first 5 mL of the filtrate. Use the filtrate as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.)
SPECIFIC TESTS CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Unit/mg of cephradine STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections PH 791: 8.09.6, in a solution containing 10 mg/mL LOSS ON DRYING 731: Dry about 100 mg in a capillarystoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 5.0% of its weight. OTHER REQUIREMENTS: It meets the requirements for Labeling under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Cephalexin RS USP Cephradine RS
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Mode: TLC Adsorbent: 0.25-mm layer of binder-free silica gel Application volume: 10 L Pre-developing solvent system: n-Hexane and tetradecane (95:5) Developing solvent system: Of 0.1 M citric acid, 0.1 M dibasic sodium phosphate, and a solution (1 in 15) of ninhydrin in acetone (60:40:1.5) Analysis Samples: Standard solution and Sample solution Place the chromatographic plate in a chamber containing the Pre-developing solvent system to a depth of 1 cm, and allow the solvent front to move the length of the plate. Apply the Samples. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, dry the plate for 10 min at 110, and examine the chromatogram. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, 0.5 M sodium acetate, 0.7 N acetic acid, and water (200:15:3:782). Pass the solution through a filter of 1-m or finer porosity. System suitability solution: 0.5 mg/mL of USP Cephradine RS and 0.5 mg/mL of USP Cephalexin RS in Mobile phase Standard solution: 0.5 mg/mL of USP Cephradine RS in Mobile phase Sample stock solution: Place NLT 5 Tablets in a high-speed glass blender jar containing a volume of water, sufficient to yield a concentration of nominally NLT 5 mg/mL of cephradine, and blend for 4 1 min. Sample solution: Nominally 0.5 mg/mL cephradine from Sample stock solution diluted with Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cephalexin and cephradine are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the cephalexin peak and the cephradine peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of label claim of cephradine (sum of cephradine and cephalexin) in each Tablet taken: Result = (rU/rS) (CS/CU) P F 100 rU rS CS CU = sum of the cephradine and cephalexin peak response from the Sample solution = sum of the cephradine and cephalexin peak response from the Standard solution = concentration of USP Cephradine RS in the Standard solution (mg/mL) = nominal concentration of cephradine from the Sample solution (mg/mL)
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for cephalexin and cephradine are about 0.8 and 1.0, respectively. ] Suitability requirements Resolution: NLT 2.0 between the cephalexin peak and the cephradine peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of label claim of cephradine (sum of cephradine and cephalexin) in constituted Oral Suspension: Result = (rU/rS) (CS/CU) P F 100 = sums of the cephradine and cephalexin peak response from the Sample solution = sums of the cephradine and cephalexin peak rS response from the Standard solution = concentration of USP Cephradine RS in the CS Standard solution (mg/mL) = nominal concentration of cephradine, from the CU Sample solution (mg/mL) P = designated potency of USP Cephradine RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%125.0% rU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For solid packaged in single-unit containers: meets the requirements DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 3.56.0, in the suspension constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 1.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cephalexin RS USP Cephradine RS
Cephradine Tablets
(Comment on this Monograph)id=m14620=Cephradine Tablets=Ca-Chl-Monos.pdf) DEFINITION Cephradine Tablets contain NLT 90.0% and NMT 120.0% of the labeled amount of cephradine, calculated as the sum of cephradine (C16H19N3O4S) and cephalexin (C16H17N3O4S). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 3 mg/mL of USP Cephradine RS in water Sample solution: 3 mg/mL of cephradine from powdered Tablets in water and filtered Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
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= designated potency of USP Cephradine RS (g/mg) F = unit conversion factor, 0.001 mg/g Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.12 N hydrochloric acid; 900 mL Apparatus 2: 75 rpm Time: 60 min Detector: UV 255 nm Standard solution: USP Cephradine RS in Medium Sample solution: Sample per Dissolution 711. Dilute filtered portions with Medium to a concentration that is similar to the Standard solution. Tolerances: NLT 85% (Q) of the labeled amount of C16H19N3O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 6.0% P
Official Monographs / Cetylpyridinium 203

IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2%, calculated on the anhydrous basis HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE: PYRIDINE: Dissolve 1 g in 10 mL of sodium hydroxide solution (1 in 10) without heating: the odor of pyridine is not immediately perceptible. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 8084, the preliminary drying treatment being omitted ACIDITY: Dissolve 500 mg in 50 mL of water, add phenolphthalein TS, and titrate with 0.020 N sodium hydroxide: NMT 2.5 mL is required for neutralization. WATER DETERMINATION, Method I 921: 4.5%5.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cetylpyridinium Chloride RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cephalexin RS USP Cephradine RS
Cetylpyridinium Chloride Lozenges

(Comment on this Monograph)id=m14790=Cetylpyridinium Chloride Lozenges=Ca-Chl-Monos.pdf) DEFINITION Cetylpyridinium Chloride Lozenges contain NLT 90.0% and NMT 125.0% of the labeled amount of C21H38ClN H2O in a suitable molded base. IDENTIFICATION PROCEDURE Eluting solvent: Alcohol and 1.2 N hydrochloric acid (7:3) Chromatographic column: Pack a pledget of fine glass wool in the base of a 10-mm 200-mm chromatographic tube. Add styrene-divinylbenzene cation-exchange resin (strong acid form), to form a uniform column 12 cm in height, and top the column with a pledget of fine glass wool. Sample solution: Dissolve the nominal equivalent of 500 g of cetylpyridinium chloride, from NLT 20 powdered Lozenges, in 50 mL of water, and immediately transfer this solution to the Chromatographic column. Discard the eluate, wash the column, successively, with 200 mL of water, 100 mL of alcohol, 100 mL of water, and 100 mL of 3 N hydrochloric acid, and discard the washings. Elute the column with 80 mL of Eluting solvent. Collect the eluate in a 100-mL volumetric flask, and dilute with the Eluting solvent to volume. Standard solution: 5 g/mL of USP Cetylpyridinium Chloride RS in Eluting solvent Spectrometric conditions Mode: UV Analytical wavelength: 225300 nm Analysis Samples: Sample solution and Standard solution Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of the Standard solution. ASSAY PROCEDURE 0.004 M sodium lauryl sulfate: Dissolve 1.15 g of sodium lauryl sulfate in 500 mL of water, add 2 mL of sulfuric acid, and dilute with water to 1000 mL.
Cetylpyridinium Chloride
(Comment on this Monograph)id=m14780=Cetylpyridinium Chloride=Ca-Chl-Monos.pdf)
358.00 C21H38ClN H2O Pyridinium, 1-hexadecyl-, chloride, monohydrate; 1-Hexadecylpyridinium chloride monohydrate [6004-24-6]. Anhydrous 339.99 [123-03-5]. DEFINITION Cetylpyridinium Chloride contains NLT 99.0% and NMT 102.0% of C21H38ClN, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 40 g/mL in water C. IDENTIFICATION TESTSGENERAL, Chloride 191: 2 mg/mL in water: a 10-mL portion of the solution meets the requirements, except that when silver nitrate TS is added, turbidity is produced rather than a curdy white precipitate. ASSAY PROCEDURE Sample solution: Dissolve 200 mg of Cetylpyridinium Chloride in 75 mL of water. Add 10 mL of chloroform, 0.4 mL of bromophenol blue solution (1 in 2000), and 5 mL of a freshly prepared solution of sodium bicarbonate (4.2 in 1000). Analysis: Titrate the Sample solution with 0.02 M sodium tetraphenylboron VS until the blue color disappears from the chloroform layer. Add the last portions of the sodium tetraphenylboron solution dropwise, agitating vigorously after each addition. Each mL of 0.02 M sodium tetraphenylboron is equivalent to 6.800 mg of C21H38ClN.
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Cetylpyridinium / Official Monographs
USP 32
Analysis: Titrate the Sample solution with 0.02 M sodium tetraphenylboron VS until the blue color disappears from the chloroform layer. Add the last portions of the sodium tetraphenylboron solution dropwise, agitating vigorously after each addition. Each mL of 0.02 M sodium tetraphenylboron is equivalent to 7.160 mg of C21H38ClN H2O. Acceptance criteria: 95.0%105.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cetylpyridinium Chloride RS
Standardization of 0.004 M sodium lauryl sulfate: Determine the molarity of the solution as follows. To a glassstoppered 100-mL cylinder, transfer 10.0 mL of 0.004 M cetylpyridinium chloride (1.432 mg/mL of USP Cetylpyridinium Chloride RS), add 5 mL of 2 N sulfuric acid, 20 mL of chloroform, and 1 mL of methyl yellow TS, and titrate with the sodium lauryl sulfate solution, with frequent vigorous shaking until the chloroform layer acquires the first permanent orange-pink color. Calculate the molarity, and restandardize before each use. [NOTESulfuric acid is included in this solution to inhibit precipitate formation. If a precipitate forms under storage, discard the solution, and prepare and standardize a fresh solution of 0.004 M sodium lauryl sulfate.] Sample solution: Dissolve the nominal equivalent of 10 mg of cetylpyridinium chloride, from 100 powdered Lozenges, in 500 mL of water. Add 5 mL of 2 N sulfuric acid, 20 mL of chloroform, and 1 mL of methyl yellow TS. Shake until the chloroform layer develops a bright yellow color. Analysis: Titrate with 0.004 M sodium lauryl sulfate, shaking thoroughly after each addition until the chloroform layer develops the first permanent orange-pink color. Each mL of 0.004 M sodium lauryl sulfate is equivalent to 1.432 mg of C21H38ClN H2O. Acceptance criteria: 90.0%125.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cetylpyridinium Chloride RS
Activated Charcoal
(Comment on this Monograph)id=m14850=Activated Charcoal=Ca-Chl-Monos.pdf) DEFINITION Activated Charcoal is the residue from the destructive distillation of various organic materials, treated to increase its adsorptive power. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Sample: 0.50 g Acceptance criteria: NMT 4.0% CHLORIDE AND SULFATE 221, Chloride Sample solution: A 10-mL portion of the filtrate obtained in the test for Reaction Acceptance criteria: Sample solution shows no more chloride than is contained in 1.5 mL of 0.020 N hydrochloric acid (NMT 0.2%). CHLORIDE AND SULFATE 221, Sulfate Sample solution: A 10-mL portion of the filtrate obtained in the test for Reaction Acceptance criteria: Sample solution shows no more sulfate than is contained in 1.0 mL of 0.020 N sulfuric acid (NMT 0.2%). SULFIDE Sample: 0.50 g Analysis: Place in a small conical flask, add 20 mL of water and 5 mL of hydrochloric acid, and boil gently. Acceptance criteria: The escaping vapors do not darken paper moistened with lead acetate TS. CYANOGEN COMPOUNDS Sample: 5 g Analysis: Add to 50 mL of water and 2 g of tartaric acid in a distilling flask connected to a condenser provided with a tightly fitting adapter, the end of which dips below the surface of a mixture of 2 mL of 1 N sodium hydroxide and 10 mL of water, contained in a small flask surrounded by ice. Heat the mixture in the distilling flask to boiling, and distill about 25 mL. Dilute the distillate with water to 50 mL, and mix. To 25 mL of the diluted distillate add 12 drops of ferrous sulfate TS, heat the mixture almost to boiling, cool, and add 1 mL of hydrochloric acid. Acceptance criteria: No blue color is produced. HEAVY METALS 231 Sample: 1.0 g Analysis: Boil with a mixture of 20 mL of 3 N hydrochloric acid and 5 mL of bromine TS for 5 min, filter, and wash the charcoal and the filter with 50 mL of boiling water. Evaporate the filtrate and washing to dryness, and to the residue add 1 mL of 1 N hydrochloric acid, 20 mL of water, and 5 mL of sulfurous acid. Boil the solution until all of the sulfur dioxide is expelled, filter if necessary, and dilute with
Cetylpyridinium Chloride Topical Solution

(Comment on this Monograph)id=m14800=Cetylpyridinium Chloride Topical Solution=Ca-Chl-Monos.pdf) DEFINITION Cetylpyridinium Chloride Topical Solution contains NLT 95.0% and NMT 105.0% of the labeled amount of C21H38ClN H2O. IDENTIFICATION A. PROCEDURE Standard solution: 40 g/mL of USP Cetylpyridinium Chloride RS in water Sample solution: 40 g/mL of cetylpyridinium chloride from Topical solution diluted with water Analysis Samples: Standard solution and Sample solution Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths. B. IDENTIFICATION TESTSGENERAL, Chloride 191 Sample solution: Evaporate on a steam bath a volume of Topical Solution equivalent to 500 mg of cetylpyridinium chloride from Topical Solution to one-half of its original volume. Acceptance criteria: A 10-mL portion of the Sample solution meets the requirements, except that a turbidity is produced, rather than a curdy white precipitate, when the silver nitrate TS is added. ASSAY PROCEDURE Sample solution: Add a volume of Topical Solution nominally equivalent to 150 mg of cetylpyridinium chloride, to 75 mL of water. Add 10 mL of chloroform, 0.4 mL of bromophenol blue solution (1 in 2000), and 5 mL of a freshly prepared solution of sodium bicarbonate (4.2 in 1000).
USP 32
water to 50 mL. To 20 mL of the solution add water to make 25 mL. Acceptance criteria: NMT 50 ppm Organic Impurities PROCEDURE 1: UNCARBONIZED CONSTITUENTS Sample: 0.25 g Analysis: Boil with 10 mL of 1 N sodium hydroxide for 5 s, and filter. Acceptance criteria: The filtrate is colorless. PROCEDURE 2: ACID-SOLUBLE SUBSTANCES Sample: 1.0 g Analysis: Boil with a mixture of 20 mL of water and 5 mL of hydrochloric acid for 5 min, filter into a tared porcelain crucible, and wash the residue with 10 mL of hot water, adding the washing to the filtrate. To the combined filtrate and washing add 1 mL of sulfuric acid, evaporate to dryness, and ignite to constant weight. Acceptance criteria: The residue weighs NMT 35 mg (NMT 3.5%). SPECIFIC TESTS ADSORPTIVE POWER Alkaloids Sample: 1 g of Activated Charcoal, previously dried at 120 for 4 h Analysis: Shake with a solution of 100 mg of strychnine sulfate in 50 mL of water for 5 min, and pass through a dry filter, rejecting the first 10 mL of the filtrate. To a 10mL portion of the subsequent filtrate add 1 drop of hydrochloric acid and 5 drops of mercuricpotassium iodide TS. Acceptance criteria: No turbidity is produced. Dyes Sample: 250 mg Analysis: Pipet 50 mL of methylene blue solution (1 in 1000) into each of two glass-stoppered, 100-mL flasks. Add Sample to one of the flasks, insert the stopper in the flask, and shake for 5 min. Pass the contents of each flask through a dry filter, rejecting the first 20 mL of each filtrate. Pipet 25-mL portions of the remaining filtrates into two 250-mL volumetric flasks. Add to each flask 50 mL of sodium acetate solution (1 in 10), mix, and add from a buret 35.0 mL of 0.1 N iodine VS, swirling the mixture during the addition. Insert the stoppers in the flasks, and allow them to stand for 50 min, shaking them vigorously at 10-min intervals. Dilute each mixture with water to volume, mix, allow to stand for 10 min, and pass through dry filters, rejecting the first 30 mL of each filtrate. Titrate the excess iodine in a 100-mL aliquot of each subsequent filtrate with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as the endpoint is approached. Calculate the mL of 0.1 N iodine consumed in each titration. Acceptance criteria: The difference between the two volumes is NLT 0.7 mL. MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Salmonella species and Escherichia coli. REACTION Sample: 3.0 g Analysis: Boil with 60 mL of water for 5 min, allow to cool, restore the original volume by the addition of water, and filter. Acceptance criteria: The filtrate is colorless and is neutral to litmus. LOSS ON DRYING 731: Dry it at 120 for 4 h: it loses NMT 15.0% of its weight.
Official Monographs / Chloral 205

Chloral Hydrate
(Comment on this Monograph)id=m14960=Chloral Hydrate=Ca-Chl-Monos.pdf)
C2H3Cl3O2 1,1-Ethanediol, 2,2,2-trichloro-; Chloral hydrate [302-17-0].
165.40
DEFINITION Chloral Hydrate contains NLT 99.5% and NMT 102.5% of C2H3Cl3O2. IDENTIFICATION PROCEDURE Sample solution: 10 mL of a 0.1 mg/mL chloral hydrate solution Analysis: To the Sample solution add 10 mL of 1ethylquinaldinium iodide solution (15 in 1000), which has been passed through a 0.45-m filter. Add 60 mL of isopropyl alcohol, 5 mL of an aqueous 0.1 M monoethanolamine solution, and 15 mL of water. Heat in a water bath at 60 for 15 min. Acceptance criteria: A blue color develops. ASSAY PROCEDURE Sample: 4 g of Chloral Hydrate Analysis: Dissolve in 10 mL of water, add 30.0 mL of 1 N sodium hydroxide VS, and allow the mixture to stand for 2 min. Add a few drops of phenolphthalein TS and titrate the residual alkali at once with 1 N sulfuric acid VS. Each mL of 1 N sodium hydroxide corresponds to 165.4 mg of C2H3Cl3O2. Acceptance criteria: 99.5%102.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221: To a solution (1 in 10) in alcohol add a few drops of silver nitrate TS: any opalescence produced does not exceed that of a control containing 0.10 mL of 0.020 N hydrochloric acid (0.007%). READILY CARBONIZABLE SUBSTANCES TEST 271: Shake 500 mg, at intervals of 5 min during 1 h, with 5 mL of sulfuric acid TS in a glass-stoppered cylinder that previously has been rinsed with sulfuric acid TS, and transfer the mixture to a comparison vessel: the mixture has no more color than Matching Fluid P. SPECIFIC TESTS ACIDITY: A solution (1 in 20) in alcohol does not at once redden moistened blue litmus paper.
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Chloral / Official Monographs
USP 32
Analysis: To 10 mL of Sample solution add 10 mL of 1ethylquinaldinium iodide solution (15 in 1000), which has been passed through a 0.45-m filter. Add 60 mL of isopropyl alcohol, 5 mL of an aqueous 0.1 M monoethanolamine solution, and 15 mL of water. Mix, and heat in a water bath at 60 for 15 min. Acceptance criteria: A blue color develops. ASSAY PROCEDURE Sample solution: 25.0 mL of Oral Solution Analysis: Transfer Sample solution to a 250-mL conical flask with the aid of several portions of water. Add 30.0 mL of 1 N sodium hydroxide VS, and mix. After the mixture has stood for 2 min, add 5 drops of phenolphthalein TS, and immediately titrate the excess sodium hydroxide with 1 N sulfuric acid VS. Transfer 5.0 mL of Oral Solution to a second 250-mL conical flask with the aid of several portions of water. Add 10 drops of phenolphthalein TS, and titrate with 0.1 N sodium hydroxide VS. Calculate the weight, in mg, of C2H3Cl3O2 in the amount of Oral Solution taken by the first titration: Result = 165.4 [A (0.5 B)] A = volume of 1 N sodium hydroxide VS consumed B = volume of 0.1 N sodium hydroxide VS consumed Acceptance criteria: 95.0%110.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.
Chloral Hydrate Capsules

(Comment on this Monograph)id=m14970=Chloral Hydrate Capsules=Ca-Chl-Monos.pdf) DEFINITION Chloral Hydrate Capsules contain NLT 95.0% and NMT 110.0% of the labeled amount of C2H3Cl3O2. IDENTIFICATION PROCEDURE Sample solution: 0.1 mg/mL of chloral hydrate from Capsule contents diluted with water Analysis: To 10 mL of Sample solution add 10 mL of 1ethylquinaldinium iodide solution (15 in 1000), which has been passed through a 0.45-m filter. Add 60 mL of isopropyl alcohol, 5 mL of an aqueous 0.1 M monoethanolamine solution, and 15 mL of water. Mix, and heat in a water bath at 60 for 15 min. Acceptance criteria: A blue color develops. ASSAY PROCEDURE Sample: Number of Capsules, nominally equivalent to 2.5 g of chloral hydrate, in a glass-stoppered, 250-mL flask Analysis: Add 25 mL of water, insert the stopper in the flask, and heat on a steam bath with frequent swirling until the Capsules are dissolved. Cool to room temperature, add 25 mL of neutralized alcohol and 20.0 mL of 1 N sodium hydroxide VS, mix, and allow the mixture to stand for 4 min. Add phenolphthalein TS and titrate the excess alkali with 1 N sulfuric acid VS. Each mL of 1 N sodium hydroxide is equivalent to 165.4 mg of C2H3Cl3O2. Acceptance criteria: 95.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 500 mL Apparatus 2: 50 rpm Time: 15 min Analysis: Place 1 Capsule in each vessel, and allow the Capsule to sink to the bottom of the vessel before starting rotation of the blade. Observe the Capsules, and record the time taken for each capsule shell to rupture. Tolerances: The requirements are met if all of the Capsules tested rupture in NMT 15 min. If 1 or 2 of the Capsules rupture in more than 15 but NMT 30 min, repeat the test on 12 additional Capsules. NMT 2 of the total of 18 Capsules tested rupture in more than 15 but NMT 30 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, preferably at a controlled room temperature.
Chlorambucil
(Comment on this Monograph)id=m15040=Chlorambucil=CaChl-Monos.pdf)
304.21 C14H19Cl2NO2 Benzenebutanoic acid, 4-[bis(2-chloroethyl)amino]-; 4-[p-[Bis(2-chloroethyl)amino]phenyl]butyric acid [305-03-3]. DEFINITION Chlorambucil contains NLT 98.0% and NMT 101.0% of C14H19Cl2NO2, calculated on the anhydrous basis. [CAUTIONGreat care should be taken to prevent inhaling particles of Chlorambucil and exposing the skin to it.] IDENTIFICATION A. INFRARED ABSORPTION 197S: A solution (1 in 125), carbon disulfide, 1-mm cell B. PROCEDURE Sample solution: Dissolve 50 mg of chlorambucil in acetone and dilute with water to 10 mL. Analysis: To the Sample solution add 1 drop of 2 N sulfuric acid, then add 4 drops of silver nitrate TS. Acceptance criteria: No opalescence is observed immediately (absence of chloride ion). Warm the solution on a steam bath: opalescence develops (presence of ionizable chlorine).
Chloral Hydrate Oral Solution

(Comment on this Monograph)id=m14990=Chloral Hydrate Oral Solution=Ca-Chl-Monos.pdf) DEFINITION Chloral Hydrate Oral Solution contains NLT 95.0% and NMT 110.0% of the labeled amount of chloral hydrate (C2H3Cl3O2). IDENTIFICATION PROCEDURE Sample solution: Equivalent to 0.1 mg/mL of chloral hydrate from Oral solution diluted with water
USP 32
ASSAY PROCEDURE Sample: 200 mg of Chlorambucil Analysis: Dissolve in 10 mL of acetone, add 10 mL of water, and titrate with 0.1 N sodium hydroxide VS, using phenolphthalein TS as the indicator. Each mL of 0.1 N sodium hydroxide is equivalent to 30.42 mg of C14H19Cl2NO2. Acceptance criteria: 98.0%101.0% SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 6569 WATER DETERMINATION, Method I 921: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorambucil RS
Official Monographs / Chlorambucil 207

Mode: LC Detector: UV 254 nm Column: 25- or 30-cm 2-mm stainless steel column, packed with spherical silica microbeads, 510 m in diameter Flow rate: Capable of giving the required resolution and a suitable elution time Injection size: 1012 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0, Standard solution Relative standard deviation: The response factor for 68 injections does not vary by more than 2.0%. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H19Cl2NO2 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100
Chlorambucil Tablets
(Comment on this Monograph)id=m15070=Chlorambucil Tablets=Ca-Chl-Monos.pdf) DEFINITION Chlorambucil Tablets contain NLT 85.0% and NMT 110.0% of the labeled amount of C14H19Cl2NO2. IDENTIFICATION INFRARED ABSORPTION 197S Sample: Shake a quantity of finely powdered Tablets, equivalent to 16 mg of chlorambucil, with 20 mL of carbon disulfide. Filter, evaporate to dryness, and dissolve the residue in 2 mL of carbon disulfide. Cell: 1 mm ASSAY PROCEDURE Mobile phase: Mix 500 mL of alcohol with 1.0 mL of glacial acetic acid. Dilute with water to 1 L. Internal standard solution: 0.4 mg/mL of USP Propylparaben RS in alcohol Standard stock solution: 1 mg/mL of USP Chlorambucil RS in alcohol Standard solution: Transfer 2.0 mL of the Standard stock solution to a 100-mL volumetric flask containing 50 mL of alcohol and, while gently swirling, add 5.0 mL of 0.1 N hydrochloric acid and 2.0 mL of Internal standard solution. Dilute with alcohol to volume. Sample solution: Transfer the nominal equivalent of 2 mg of chlorambucil from finely powdered Tablets (NLT 20) to a 100-mL volumetric flask containing 50 mL of alcohol and, while gently swirling, add 5.0 mL of 0.1 N hydrochloric acid and 2.0 mL of Internal standard solution. Sonicate for 5 min, and dilute with alcohol to volume. Filter through a mediumporosity, sintered-glass filtering funnel, maintaining reduced pressure for the minimum necessary time to avoid solvent loss of evaporation. Chromatographic system (See Chromatography 621, System Suitability.)
= peak response ratio of chlorambucil to internal standard from the Sample solution = peak response ratio of chlorambucil to internal RS standard from the Standard solution = concentration of the Standard solution (g/mL) CS = nominal concentration of chlorambucil in the CU Sample solution (g/mL) Acceptance criteria: 85.0%110.0% PERFORMANCE TESTS DISINTEGRATION 701 Analysis: Place 1 Tablet in each of the six tubes of the basket, and if the Tablet has a soluble external coating, immerse the basket in water at room temperature for 5 min. Operate the apparatus, using simulated gastric fluid TS maintained at 37 2 as the immersion fluid. After 30 min of operation in simulated gastric fluid TS, lift the basket from the fluid, and observe the Tablets. If the Tablets have not disintegrated completely, substitute simulated intestinal fluid TS maintained at 37 2 as the immersion fluid, and continue the test for a total period of time equal to 45 min, including previous exposure to water and simulated gastric fluid TS. Lift the basket from the fluid, and observe the Tablets. Acceptance criteria: All of the Tablets have disintegrated completely. If 1 or 2 Tablets fail to disintegrate completely, repeat the test on 12 additional Tablets: NLT 16 of the total of 18 Tablets tested disintegrate completely. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve coated Tablets in wellclosed containers. Preserve uncoated Tablets in well-closed, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorambucil RS USP Propylparaben RS
RU
208
Chloramphenicol / Official Monographs
USP 32
Sample solution: 10 mg/mL of Chloramphenicol in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Developing solvent system: Chloroform, methanol, and glacial acetic acid (79:14:7) Analysis Samples: Standard solution B, Standard solution C, and Sample solution Proceed as directed under General Chapter. Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, air-dry, and examine under short-wavelength UV light. Acceptance criteria: Any spot other than the principal spot of the Sample solution does not exceed in size or intensity the principal spot of Standard solution B (1%); and the sum of the impurities represented by all of the spots other than the principal spot, based on a comparison of the intensities of such spots with the intensities of the principal spots of Standard solutions B and C, does not exceed 2%. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 149153 OPTICAL ROTATION, Specific Rotation 781S: +17.0 to +20.0 Sample solution: 50 mg/mL, undried, in dehydrated alcohol CRYSTALLINITY 695: Meets the requirements PH 791: 4.57.5, in an aqueous suspension containing 25 mg/mL BACTERIAL ENDOTOXINS TEST 85: Where Chloramphenicol is intended for use in preparing injectable dosage forms, it contains NMT 0.2 USP Endotoxin Unit/mg of chloramphenicol. STERILITY TESTS 71: Where the label states that Chloramphenicol is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration, except to use 1 g of solid specimen. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable or other sterile dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable or other sterile dosage forms. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Endotoxin RS
Chloramphenicol
(Comment on this Monograph)id=m15120=Chloramphenicol=Ca-Chl-Monos.pdf)
323.13 C11H12Cl2N2O5 Acetamide, 2,2-dichloro-N-[2-hydroxy-1-(hydroxymethyl)-2-(4nitrophenyl)ethyl]-, [R-(R*,R*)]-; D-threo-(-)-2,2-Dichloro-N-[-hydroxy--(hydroxymethyl)-pnitrophenethyl]acetamide [56-75-7]. DEFINITION Chloramphenicol contains NLT 97.0% and NMT 103.0% of C11H12Cl2N2O5. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard solution: 80 g/mL of USP Chloramphenicol RS in Mobile phase. Filter. Sample solution: 80 g/mL of chloramphenicol in Mobile phase. Pass a portion of the solution through a filter having a porosity of 0.5 m or finer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in the portion of Chloramphenicol taken: Result = (rU/rS) (CS/CU) 100 rU = peak response from the Sample solution = peak response from the Standard solution rS CS = concentration of the Standard solution (g/mL) = concentration of the Sample solution (g/mL) CU Acceptance criteria: 97.0%103.0% IMPURITIES Organic Impurities PROCEDURE Standard solution A: 10 mg/mL of USP Chloramphenicol RS in methanol Standard solution B: 100 g/mL of USP Chloramphenicol RS from Standard solution A in methanol Standard solution C: 50 g/mL of USP Chloramphenicol RS from Standard solution A in methanol
Chloramphenicol Capsules
(Comment on this Monograph)id=m15130=Chloramphenicol Capsules=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Capsules contain NLT 90.0% and NMT 120.0% of the labeled amount of chloramphenicol (C11H12Cl2N2O5). IDENTIFICATION A The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
USP 32
ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard solution: Transfer 25 mg of USP Chloramphenicol RS to a 200-mL volumetric flask, add 10 mL of water, and heat on a steam bath until completely dissolved. Cool to room temperature, dilute with Mobile phase to volume, and pass through a 0.5-m or finer porosity filter. Sample stock solution: Transfer a number of Capsules, nominally equivalent to 2500 mg of chloramphenicol, to a 1000-mL volumetric flask, add 100 mL of water, and heat on a steam bath until the Capsules have disintegrated. Add 300 mL of water, and heat on a steam bath for 20 min, with occasional mixing. Cool to room temperature, and dilute with water to volume. Sample solution: Transfer 5.0 mL of the Sample stock solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Pass a portion of the solution through a 0.5-m or finer porosity filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in each Capsule taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (mg/mL) = nominal concentration of chloramphenicol in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm Time: 30 min Standard solution: USP Chloramphenicol RS at a known concentration in Medium Sample solution: Filtered sample per Dissolution 711, diluted with medium as necessary. Spectrometric conditions Mode: UV Analytical wavelength: 278 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 85% (Q) of the labeled amount of C11H12Cl2N2O5 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
Official Monographs / Chloramphenicol 209
Chloramphenicol Cream
(Comment on this Monograph)id=m15134=Chloramphenicol Cream=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Cream contains NLT 90.0% and NMT 130.0% of the labeled amount of C11H12Cl2N2O5. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard stock solution: Transfer 40 mg of USP Chloramphenicol RS to a 100-mL volumetric flask, dissolve in methanol, and dilute with methanol to volume. Standard solution: 10.0 mL of Standard stock solution diluted with Mobile phase to 50.0 mL. Pass through a 0.5m or finer porosity filter, and use the clear filtrate. Sample stock solution: Transfer Cream, equivalent to 40 mg of chloramphenicol, to a 100-mL volumetric flask. Add 80 mL of methanol, and sonicate for 10 min. Cool to room temperature, and dilute with methanol to volume. Sample solution: 10.0 mL of Sample stock solution diluted with Mobile phase to 50.0 mL. Filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in the portion of Cream taken: Result = (rU/rS) (CS/CU) 100 = peak height from the Sample solution = peak height from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of chloramphenicol in the CU Sample solution (g/mL) Acceptance criteria: 90.0%130.0% rU rS CS PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight containers. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
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USP 32
LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Endotoxin RS
Chloramphenicol Injection
(Comment on this Monograph)id=m15140=Chloramphenicol Injection=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Injection is a sterile solution of Chloramphenicol in one or more suitable solvents. It contains NLT 90.0% and NMT 115.0% of the labeled amount of C11H12Cl2N2O5. It may contain suitable buffers. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard solution: Dissolve 80 g/mL of USP Chloramphenicol RS in Mobile phase, filter, and use the clear filtrate. Sample stock solution: Transfer 200 mg of chloramphenicol from Injection to a 100-mL volumetric flask, and add Mobile phase to volume. Sample solution: 1.0 mL of Sample stock solution diluted with Mobile phase to 25.0 mL. Pass through a 0.5-m or finer porosity filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in each mL of Injection taken: Result = (rU/rS) (CS/CU) 100 = peak height from the Sample solution = peak height from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of chloramphenicol in CU the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% rU rS CS SPECIFIC TESTS PH 791: 5.08.0, in a solution diluted with water (1:1) BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.2 USP Endotoxin Unit/mg of chloramphenicol. STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration, 1 mL from each container being transferred directly to the membrane filter. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers.
Chloramphenicol Ophthalmic Ointment

(Comment on this Monograph)id=m15150=Chloramphenicol Ophthalmic Ointment=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Ophthalmic Ointment contains NLT 90.0% and NMT 130.0% of the labeled amount of C11H12Cl2N2O5. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard stock solution: 0.25 mg/mL of USP Chloramphenicol RS in methanol Standard solution: 10.0 mL of Standard stock solution diluted with Mobile phase to 25.0 mL. Pass through a 0.5m or finer porosity filter, and use the clear filtrate. Sample stock solution: 25 mg of chloramphenicol from Ophthalmic Ointment, to a suitable conical flask. Add 20 mL of cyclohexane, and sonicate for 2 min. Add 60 mL of methanol. Filter this mixture, collecting the filtrate in a 100mL volumetric flask. Wash the filter with methanol, collecting the washings in the volumetric flask. Dilute with methanol to volume. Transfer 50.0 mL of the resulting solution to a suitable round-bottom flask, and evaporate to dryness by rotating the flask under vacuum in a water bath at 35. Dissolve the residue in 50.0 mL of methanol. Sample solution: 10.0 mL of Sample stock solution diluted with Mobile phase to 25.0 mL. Pass through a 0.5-m or finer porosity filter and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in the portion of Ophthalmic Ointment taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak height from the Sample solution = peak height from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of chloramphenicol in the Sample solution (g/mL)
USP 32

STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a refrigerator until dispensed. The containers or individual cartons are sealed and tamper proof so that sterility is assured at the time of first use. LABELING: The labeling states that there is a 21-day, beyonduse period after dispensing. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements MINIMUM FILL 755: Meets the requirements METAL PARTICLES: It meets the requirements under Metal Particles in Ophthalmic Ointments 751. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
Chloramphenicol Ophthalmic Solution

(Comment on this Monograph)id=m15160=Chloramphenicol Ophthalmic Solution=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Ophthalmic Solution is a sterile solution of Chloramphenicol. It contains NLT 90.0% and NMT 130.0% of the labeled amount of C11H12Cl2N2O5. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard solution: 100 g/mL of USP Chloramphenicol RS in Mobile phase. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Sample solution: 100 g/mL of chloramphenicol from Ophthalmic Solution, in Mobile phase. Pass through a 0.5m or finer porosity filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in each mL of the Ophthalmic Solution taken: Result = (rU/rS) (CS/CU) 100 = peak height of the Sample solution = peak height of the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of chloramphenicol in the CU Sample solution (g/mL) Acceptance criteria: 90.0%130.0% SPECIFIC TESTS PH 791: 7.07.5, or for an unbuffered Opthalmic Solution or an Opthalmic Solution that is labeled for veterinary use, 3.06.0 rU rS CS
Chloramphenicol for Ophthalmic Solution

(Comment on this Monograph)id=m15161=Chloramphenicol for Ophthalmic Solution=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol for Ophthalmic Solution is a sterile, dry mixture of Chloramphenicol with or without one or more suitable buffers, diluents, and preservatives. It contains NLT 90.0% and NMT 130.0% of the labeled amount of C11H12Cl2N2O5, when constituted as directed. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard solution: 100 g/mL of USP Chloramphenicol RS in Mobile phase. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Sample solution: Transfer the contents of 1 container of Chloramphenicol for Ophthalmic Solution to a suitable container with the aid of the Mobile phase, and prepare a solution containing 100 g/mL of Chloramphenicol with the Mobile phase. Pass a portion of the solution through a 0.5m or finer porosity filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280-nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak height from the Sample solution = peak height from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of chloramphenicol in the Sample solution (g/mL)
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90.0%130.0%
USP 32
IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard solution: 100 g/mL of USP Chloramphenicol RS in Mobile phase. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Sample stock solution: 50 mg of chloramphenicol from Otic Solution to a 100-mL volumetric flask. Dilute with Mobile phase to volume. Sample solution: 10.0 mL of Sample stock solution diluted with Mobile phase to 50.0 mL. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in each mL of the Otic Solution taken: Result = (rU/rS) (CS/CU) 100 = peak height from the Sample solution = peak height from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) CU = nominal concentration of chloramphenicol in the Sample solution (g/mL) Acceptance criteria: 90.0%130.0% rU rS CS SPECIFIC TESTS PH 791: 4.08.0, when diluted with an equal volume of water STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
SPECIFIC TESTS PH 791: 7.17.5, in 5 mg/mL of chloramphenicol aqueous solution STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: If packaged in combination with a container of solvent, label it with a warning that it is not for injection. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
Chloramphenicol Oral Solution

(Comment on this Monograph)id=m15170=Chloramphenicol Oral Solution=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Oral Solution is a solution of Chloramphenicol in a suitable solvent. It contains one or more suitable buffers and preservatives. It has a potency of NLT 90.0% and NMT 120.0% of the labeled amount of C11H12Cl2N2O5. IDENTIFICATION PROCEDURE Sample solution: 20 g/mL chloramphenicol from Oral Solution diluted with water Standard solution: 20 g/mL USP Chloramphenicol RS Spectrometric conditions Mode: UV Analysis Samples: Sample solution and Standard solution Acceptance criteria: The UV spectrum of the Sample solution exhibits maxima and minima only at the same wavelength as that of the Standard solution. ASSAY PROCEDURE Sample solution: Oral Solution diluted to yield a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81. Acceptance criteria: 90.0%120.0% SPECIFIC TESTS PH 791: 5.08.5, when diluted with an equal volume of water ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that it is for veterinary use only and that it is not to be used in animals raised for food production. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
Chloramphenicol Tablets Chloramphenicol Otic Solution

(Comment on this Monograph)id=m15173=Chloramphenicol Otic Solution=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Otic Solution is a sterile solution of Chloramphenicol in a suitable solvent. It contains NLT 90.0% and NMT 130.0% of the labeled amount of C11H12Cl2N2O5. (Comment on this Monograph)id=m15183=Chloramphenicol Tablets=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol Tablets contain NLT 90.0% and NMT 120.0% of the labeled amount of C11H12Cl2N2O5.
USP 32
IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard solution: 25 mg of USP Chloramphenicol RS to a 200-mL volumetric flask. Add 10 mL of water, and heat on a steam bath until completely dissolved. Cool to room temperature, and dilute with Mobile phase to volume. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Sample stock solution: 500 mg of chloramphenicol from finely powdered Tablets (NLT 20 Tablets) to a 200-mL volumetric flask. Add 80 mL of water, and heat on a steam bath for 20 min, with occasional mixing. Cool to room temperature, and dilute with water to volume. Sample solution: 1.0 mL of Sample stock solution diluted with Mobile phase to 20.0 mL. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak height from the Sample solution = peak height from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) CU = nominal concentration of chloramphenicol in the Sample solution (g/mL) Acceptance criteria: 90.0%120.0% rU rS CS PERFORMANCE TESTS DISINTEGRATION 701 Time: 60 min UNIFORMITY OF DOSAGE UNITS 905:
Chloramphenicol and Hydrocortisone Acetate for Ophthalmic Suspension

(Comment on this Monograph)id=m15220=Chloramphenicol and Hydrocortisone Acetate for Ophthalmic Suspension=Ca-ChlMonos.pdf) DEFINITION Chloramphenicol and Hydrocortisone Acetate for Ophthalmic Suspension is a sterile, dry mixture of Chloramphenicol and Hydrocortisone Acetate with or without one or more suitable buffers, diluents, and preservatives. It contains NLT 90.0% and NMT 130.0% of the labeled amount of chloramphenicol (C11H12Cl2N2O5), and NLT 90.0% and NMT 115.0% of the labeled amount of hydrocortisone acetate (C23H32O6), when constituted as directed. IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard stock solution: 37.5 mg of USP Chloramphenicol RS and 37.5J mg of USP Hydrocortisone Acetate RS, J being the ratio of the labeled amount, in mg, of hydrocortisone acetate to the labeled amount, in mg, of chloramphenicol in the Chloramphenicol and Hydrocortisone Acetate for Ophthalmic Suspension. To a 100-mL volumetric flask, add 15 mL of water and 75 mL of methanol, sonicate for a few seconds, and dilute with methanol to volume. Standard solution: 10.0 mL of Standard stock solution diluted with Mobile phase to 50.0 mL. Pass through a 0.5m or finer porosity filter, and use the clear filtrate. Sample stock solution: Contents of an accurately counted number of containers of Chloramphenicol and Hydrocortisone Acetate for Ophthalmic Suspension, equivalent to 37.5 mg of chloramphenicol, to a 100-mL volumetric flask with the aid of 5 mL of water for each 12.5 mg of chloramphenicol contained therein. Wash each container with methanol, and add the washings to the volumetric flask. Dilute with methanol to volume. Sample solution: 10.0 mL of Sample stock solution diluted with Mobile phase to 50.0 mL. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label Tablets to indicate that they are for veterinary use only and are not to be used in animals raised for food production. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS
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USP 32
Sample stock solution: 25 mg of chloramphenicol from Chloramphenicol and Polymyxin B Sulfate Ophthalmic Ointment, to a suitable conical flask. Add 20 mL of cyclohexane, mix, and sonicate for 2 min. Add 60 mL of methanol, and mix. Filter this mixture, collecting the filtrate in a 100-mL volumetric flask. Wash the filter with methanol, collecting the washings in the volumetric flask. Dilute with methanol to volume. Transfer 50.0 mL of the resulting solution to a suitable round-bottom flask, and evaporate to dryness by rotating the flask under vacuum in a water bath at 35. Dissolve the residue in 50.0 mL of methanol. Sample solution: 10.0 mL of Sample stock solution diluted with Mobile phase to 25.0 mL. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5 m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in the portion of Ophthalmic Ointment taken: Result = (rU/rS) (CS/CU) 100 = chloramphenicol peak height from the Sample solution = chloramphenicol peak height from the Standard rS solution = concentration of USP Chloramphenicol RS in the CS Standard solution (g/mL) = nominal concentration of chloramphenicol in the CU Sample solution (g/mL) Acceptance criteria: 90.0%120.0% POLYMYXIN Analysis: Proceed as directed for polymyxin under AntibioticsMicrobial Assays 81, using a portion of Ophthalmic Ointment equivalent to 5000 Polymyxin B Units, shaken in a separator containing 50 mL of ether, and extracted with four 20-mL portions of Buffer No. 6. Combine the aqueous extracts in a 100-mL volumetric flask, dilute with Buffer No. 6 to volume, and mix. Dilute an accurately measured portion of this solution quantitatively with Buffer No. 6 to obtain a Test Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%125.0% SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements METAL PARTICLES: It meets the requirements of the test for Metal Particles in Ophthalmic Ointments 751. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Polymyxin B Sulfate RS rU
Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in each container taken: Result = (rU/rS) (CS/CU) 100 = peak height of the Sample solution = peak height of the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of chloramphenicol in CU the Sample solution (g/mL) Acceptance criteria: 90.0%130.0% Calculate the percentage of C23H32O6 in each container taken: rU rS CS Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Hydrocortisone Acetate RS in the Standard solution (g/mL) = nominal concentration of hydrocortisone acetate CU in the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS PH 791: 7.17.5, in 5 mg/mL chloramphenicol aqueous suspension STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. ADDITIONAL REQUIREMENTS LABELING: If packaged in combination with a container of solvent, label it with a warning that it is not for injection. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Hydrocortisone Acetate RS rU rS CS
Chloramphenicol and Polymyxin B Sulfate Ophthalmic Ointment

(Comment on this Monograph)id=m15240=Chloramphenicol and Polymyxin B Sulfate Ophthalmic Ointment=Ca-ChlMonos.pdf) DEFINITION Chloramphenicol and Polymyxin B Sulfate Ophthalmic Ointment contains NLT 90.0% and NMT 120.0% of the labeled amount of chloramphenicol (C11H12Cl2N2O5) and NLT 90.0% and NMT 125.0% of the labeled amount of polymyxin B. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution as obtained in the Assay for Chloramphenicol. ASSAY CHLORAMPHENICOL Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard stock solution: 0.25 mg/mL of USP Chloramphenicol RS in methanol Standard solution: 10.0 mL of Standard stock solution diluted with Mobile phase to 25.0 mL. Pass through a 0.5m or finer porosity filter, and use the clear filtrate.
USP 32

Sample solution: 10.0 mL of Sample stock solution diluted with Mobile phase to 25.0 mL. Pass through a 0.5-m or finer porosity filter, and use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 10-cm; 5m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1800 theoretical plates from the analyte peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in the portion of Ophthalmic Ointment taken: Result = (rU/rS) (CS/CU) 100 = peak response of chloramphenicol from the Sample solution rS = peak response of chloramphenicol from the Standard solution CS = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) CU = nominal concentration of chloramphenicol in the Sample solution (g/mL) Acceptance criteria: 90.0%120.0% of the labeled amount of C11H12Cl2N2O5 Calculate the percentage of C23H32O6 in the portion of Ophthalmic Ointment taken: rU Result = (rU/rS) (CS/CU) 100 = peak response of hydrocortisone acetate from the Sample solution = peak response of hydrocortisone acetate from the rS Standard solution = concentration of USP Hydrocortisone Acetate RS CS in the Standard solution (g/mL) = nominal concentration of hydrocortisone acetate CU in the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% of the labeled amount of C23H32O6 rU SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements MINIMUM FILL 755: Meets the requirements METAL PARTICLES: It meets the requirements of the test for Metal Particles in Ophthalmic Ointments 751. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Hydrocortisone Acetate RS USP Polymyxin B Sulfate RS
Chloramphenicol, Polymyxin B Sulfate, and Hydrocortisone Acetate Ophthalmic Ointment

(Comment on this Monograph)id=m15250=Chloramphenicol, Polymyxin B Sulfate, and Hydrocortisone Acetate Ophthalmic Ointment=Ca-Chl-Monos.pdf) DEFINITION Chloramphenicol, Polymyxin B Sulfate, and Hydrocortisone Acetate Ophthalmic Ointment contains NLT 90.0% and NMT 120.0% of the labeled amount of chloramphenicol (C11H12Cl2N2O5); NLT 90.0% and NMT 125.0% of the labeled amount of polymyxin B; and NLT 90.0% and NMT 115.0% of the labeled amount of hydrocortisone acetate (C23H32O6). IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay for chloramphenicol and hydrocortisone acetate. ASSAY POLYMYXIN Analysis: Proceed as directed for polymyxin under AntibioticsMicrobial Assays 81, using a portion of Ophthalmic Ointment equivalent to 5000 Polymyxin B Units, shaken in a separator containing 50 mL of ether and extracted with four 20-mL portions of Buffer No. 6. Combine the aqueous extracts in a 100-mL volumetric flask, dilute with Buffer No. 6 to volume, and mix. Dilute a portion of this solution quantitatively with Buffer No. 6 to obtain a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%125.0% of the labeled amount of polymyxin B CHLORAMPHENICOL AND HYDROCORTISONE ACETATE Mobile phase: Methanol, glacial acetic acid, and water (45:0.1:55) Standard stock solution: 25 mg of USP Chloramphenicol RS and 25J mg of USP Hydrocortisone Acetate RS to a 100mL volumetric flask, dissolve in methanol, and dilute with methanol to volume [NOTEJ is the ratio of the labeled amount, in mg, of hydrocortisone acetate to the labeled amount, in mg, of chloramphenicol/g of Ophthalmic Ointment.] Standard solution: 10.0 mL of Standard stock solution diluted with Mobile phase to 25.0 mL. Pass through a 0.5m or finer porosity filter, and use the clear filtrate. Sample stock solution: 25 mg of chloramphenicol from Chloramphenicol, Polymyxin B Sulfate, and Hydrocortisone Acetate Ophthalmic Ointment, to a suitable conical flask. Add 20 mL of cyclohexane, mix, and sonicate for 2 min. Add 60 mL of methanol, and mix. Filter this mixture, collecting the filtrate in a 100-mL volumetric flask. Wash the filter with methanol, collecting the washings in the volumetric flask. Dilute with methanol to volume. Transfer 50.0 mL of the resulting solution to a suitable round-bottom flask, and evaporate to dryness by rotating the flask under vacuum in a water bath at 35. Dissolve the residue in 50.0 mL of methanol.
USP 32
Chloramphenicol and Prednisolone Ophthalmic Ointment Chloramphenicol Palmitate Chloramphenicol Palmitate Oral Suspension Chloramphenicol Sodium Succinate Chloramphenicol Sodium Succinate for Injection Chlordiazepoxide Chlordiazepoxide Tablets Chlordiazepoxide and Amitriptyline Hydrochloride Tablets Chlordiazepoxide Hydrochloride Chlordiazepoxide Hydrochloride Capsules Chlordiazepoxide Hydrochloride for Injection Chlordiazepoxide Hydrochloride and Clidinium Bromide Capsules Chlorhexidine Gluconate Oral Rinse Chlorhexidine Gluconate Solution Chlorophyllin Copper Complex Sodium Chloroprocaine Hydrochloride Chloroprocaine Hydrochloride Injection Chloroquine Chloroquine Hydrochloride Injection Chloroquine Phosphate Chloroquine Phosphate Tablets Chlorothiazide Chlorothiazide Oral Suspension Chlorothiazide Tablets Chlorothiazide Sodium for Injection Chloroxylenol Chlorpheniramine Maleate Chlorpheniramine Maleate Extended-Release Capsules Chlorpheniramine Maleate Injection Chlorpheniramine Maleate Oral Solution Chlorpheniramine Maleate Tablets Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride Extended-Release Capsules Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride Extended-Release Tablets Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Extended-Release Capsules Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Oral Solution Chlorpromazine Chlorpromazine Suppositories Chlorpromazine Hydrochloride Chlorpromazine Hydrochloride Oral Concentrate Chlorpromazine Hydrochloride Injection
MONOGRAPH LIST /
Chlorpromazine Hydrochloride Syrup Chlorpromazine Hydrochloride Tablets Chlorpropamide Chlorpropamide Tablets Chlortetracycline Bisulfate Chlortetracycline and Sulfamethazine Bisulfates Soluble Powder Chlortetracycline Hydrochloride Chlortetracycline Hydrochloride Ointment Chlortetracycline Hydrochloride Ophthalmic Ointment Chlortetracycline Hydrochloride Soluble Powder Chlortetracycline Hydrochloride Tablets Chlorthalidone Chlorthalidone Tablets Chlorzoxazone Chlorzoxazone Tablets Cholecalciferol Cholecalciferol Solution Cholestyramine Resin Cholestyramine for Oral Suspension Chromic Chloride Chromic Chloride Injection Sodium Chromate Cr 51 Injection Chromium Cr 51 Edetate Injection Chymotrypsin Chymotrypsin for Ophthalmic Solution Ciclopirox Ciclopirox Olamine Ciclopirox Olamine Cream Ciclopirox Olamine Topical Suspension Cilastatin Sodium Cilostazol Cilostazol Tablets Cimetidine Cimetidine Tablets Cimetidine Hydrochloride Cimetidine Injection Cimetidine in Sodium Chloride Injection Cinoxacin Cinoxacin Capsules Ciprofloxacin Ciprofloxacin Hydrochloride Ciprofloxacin and Dexamethasone Otic Suspension
/ MONOGRAPH LIST
Clobetasol Propionate Topical Solution Clocortolone Pivalate Clocortolone Pivalate Cream Clofazimine Clofazimine Capsules Clofibrate Clofibrate Capsules Clomiphene Citrate Clomiphene Citrate Tablets Clomipramine Hydrochloride Clomipramine Hydrochloride Capsules Clonazepam Clonazepam Oral Suspension Clonazepam Tablets Clonidine Clonidine Hydrochloride Clonidine Hydrochloride Tablets Clonidine Hydrochloride and Chlorthalidone Tablets Clonidine Transdermal System Clopidogrel Bisulfate Clopidogrel Tablets Clorazepate Dipotassium Clorazepate Dipotassium Tablets Clorsulon Clotrimazole Clotrimazole Cream Clotrimazole Lotion Clotrimazole Lozenges Clotrimazole Topical Solution Clotrimazole Vaginal Inserts
USP 32
Ciprofloxacin Injection Ciprofloxacin Ophthalmic Ointment Ciprofloxacin Ophthalmic Solution Ciprofloxacin Tablets Cisplatin Cisplatin for Injection Citalopram Hydrobromide Citalopram Tablets Anhydrous Citric Acid Citric Acid Monohydrate Citric Acid, Magnesium Oxide, and Sodium Carbonate Irrigation Cladribine Clarithromycin Clarithromycin for Oral Suspension Clarithromycin Tablets Clarithromycin Extended-Release Tablets Clavulanate Potassium Clemastine Fumarate Clemastine Fumarate Tablets Clidinium Bromide Clindamycin Hydrochloride Clindamycin Hydrochloride Capsules Clindamycin Hydrochloride Oral Solution Clindamycin Palmitate Hydrochloride Clindamycin Palmitate Hydrochloride for Oral Solution Clindamycin Phosphate Clindamycin Phosphate Vaginal Cream Clindamycin Phosphate Gel Clindamycin Injection Clindamycin for Injection Clindamycin Phosphate Topical Solution Clindamycin Phosphate Topical Suspension Clindamycin Phosphate Vaginal Inserts Clioquinol Clioquinol Cream Clioquinol Ointment Compound Clioquinol Topical Powder Clioquinol and Hydrocortisone Cream Clioquinol and Hydrocortisone Ointment Clobetasol Propionate Clobetasol Propionate Cream Clobetasol Propionate Ointment
Clotrimazole and Betamethasone Dipropionate Cream Cloxacillin Benzathine Cloxacillin Benzathine Intramammary Infusion Cloxacillin Sodium Cloxacillin Sodium Capsules Cloxacillin Sodium Intramammary Infusion Cloxacillin Sodium for Oral Solution Clozapine Clozapine Tablets Coal Tar Coal Tar Ointment Coal Tar Topical Solution Cyanocobalamin Co 57 Capsules
USP 32
Cyanocobalamin Co 57 Oral Solution Cyanocobalamin Co 58 Capsules Cocaine Cocaine Hydrochloride Cocaine Hydrochloride Tablets for Topical Solution Cocaine and Tetracaine Hydrochlorides and Epinephrine Topical Solution Coccidioidin Cod Liver Oil Codeine Codeine Phosphate Codeine Phosphate Injection Codeine Phosphate Tablets Codeine Sulfate Codeine Sulfate Tablets Colchicine Colchicine Injection Colchicine Tablets Colestipol Hydrochloride Colestipol Hydrochloride for Oral Suspension Colestipol Hydrochloride Tablets Colistimethate Sodium Colistimethate for Injection Colistin Sulfate Colistin Sulfate for Oral Suspension Colistin and Neomycin Sulfates and Hydrocortisone Acetate Otic Suspension Collodion Flexible Collodion Colloidal Oatmeal Copper Gluconate Corticotropin Injection Corticotropin for Injection Repository Corticotropin Injection Corticotropin Zinc Hydroxide Injectable Suspension Cortisone Acetate Cortisone Acetate Injectable Suspension Cortisone Acetate Tablets Purified Cotton Cromolyn Sodium
MONOGRAPH LIST /
Cromolyn Sodium Inhalation Powder Cromolyn Sodium Inhalation Solution Cromolyn Sodium Nasal Solution Cromolyn Sodium Ophthalmic Solution Crotamiton Crotamiton Cream Cupric Chloride Cupric Chloride Injection Cupric Sulfate Cupric Sulfate Injection Cyanocobalamin Cyanocobalamin Injection Cyclandelate Cyclizine Hydrochloride Cyclizine Hydrochloride Tablets Cyclobenzaprine Hydrochloride Cyclobenzaprine Hydrochloride Tablets Cyclopentolate Hydrochloride Cyclopentolate Hydrochloride Ophthalmic Solution Cyclophosphamide Cyclophosphamide for Injection Cyclophosphamide Tablets Cyclopropane Cycloserine Cycloserine Capsules Cyclosporine Cyclosporine Capsules Cyclosporine Injection Cyclosporine Oral Solution Cyproheptadine Hydrochloride Cyproheptadine Hydrochloride Oral Solution Cyproheptadine Hydrochloride Tablets Cyromazine Cysteine Hydrochloride Cysteine Hydrochloride Injection Cytarabine Cytarabine for Injection
USP 32
Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Resolution: The peak for chloramphenicol obtained from the Ophthalmic Ointment, at a retention time corresponding to that of the peak obtained from the USP Reference Standard, exhibits baseline separation from the adjacent prednisolone peak. Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 in the portion of Ophthalmic Ointment taken: Result = (rU/rS) (CS/CU) 100 = peak height of the Sample solution = peak height of the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (mg/mL) CU = nominal concentration of chloramphenicol in the Sample solution (mg/mL) Acceptance criteria: 90.0%130.0% PREDNISOLONE Solution A, Solution B, and Mobile phase: Proceed as directed in the Assay for Chloramphenicol. Standard solution: 0.2 mg/mL of USP Prednisolone RS in Solution A Sample solution: 2.0 mg of Prednisolone from Ophthalmic Ointment, to a screw-capped test tube. Add 10 mL of nheptane, and shake by mechanical means until the substance is dissolved. Add 10.0 mL of Solution A, and shake by mechanical means for 30 s. Allow the layers to separate, and carefully remove the upper phase. Centrifuge the lower phase for 15 min, and use the clear portion. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 15 L System suitability Sample: Standard solution Suitability requirements Resolution: The peak for prednisolone obtained from the Ophthalmic Ointment, at a retention time corresponding to that of the peak obtained from the USP Reference Standard exhibits baseline separation from the adjacent chloramphenicol peak. Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution. Calculate the percentage of C21H28O5 in the portion of Ophthalmic Ointment taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of the Sample solution rU rS CS
Chloramphenicol and Prednisolone Ophthalmic Ointment

(Comment on this Monograph)id=m15280=Chloramphenicol and Prednisolone Ophthalmic Ointment=Chl-Cy-Monos.pdf) DEFINITION Chloramphenicol and Prednisolone Ophthalmic Ointment contains NLT 90.0% and NMT 130.0% of the labeled amount of chloramphenicol (C11H12Cl2N2O5), and NLT 90.0% and NMT 115.0% of the labeled amount of prednisolone (C21H28O5). IDENTIFICATION A. PROCEDURE Sample: 20 mg of chloramphenicol from Ophthalmic Ointment Analysis: Transfer the Sample to a screw-capped test tube, add 5 mL of 5 N sodium hydroxide and 2 mL of pyridine, and shake. Place the tube in a water bath at 50 for 20 min. Acceptance criteria: A reddish brown color develops in the pyridine layer. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chloramphenicol. C. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 0.5 mg/mL of USP Prednisolone RS in chloroform Sample solution: 1.5 mg of prednisolone from Ophthalmic Ointment, to a screw-capped test tube. Add 10 mL of methylene chloride, and shake to disperse. Heat at 60 for 15 min, and allow to cool while shaking for 30 min. Allow to separate, draw off the upper ointment layer, and retain the lower methylene chloride layer. Application volume: 0.4 mL Developing solvent system: Chloroform and acetone (4:1) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow each portion to dry before adding the next portion to the same spot. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY CHLORAMPHENICOL Solution A: Methanol and water (4:1) Solution B: 1.36 mg/mL of sodium acetate trihydrate in water. Adjust with glacial acetic acid to a pH of 4.0. Mobile phase: Methanol and Solution B (1:1) Standard solution: 0.3 mg/mL of USP Chloramphenicol RS in Solution A Sample solution: 3.0 mg of chloramphenicol from Ophthalmic Ointment, to a screw-capped test tube. Add 10 mL of n-heptane, and shake by mechanical means until the substance is dissolved. Add 10.0 mL of Solution A, and shake by mechanical means for 30 s. Allow the layers to separate, and carefully remove the upper phase. Centrifuge the lower phase for 15 min, and use the clear portion. Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
= peak response of the Standard solution = concentration of USP Prednisolone RS in the Standard solution (mg/mL) = nominal concentration of prednisolone in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements METAL PARTICLES: It meets the requirements of the test for Metal Particles in Ophthalmic Ointments 751. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Prednisolone RS rS CS

Relative standard deviation: NMT 0.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage, in g, of C11H12Cl2N2O5 equivalent in each mg of specimen taken: Result = (rU/rS) (CS/CU) P 100 = peak response of chloramphenicol palmitate from the Sample solution = peak response of chloramphenicol palmitate rS from the Standard solution CS = concentration of USP Chloramphenicol Palmitate RS in the Standard solution (mg/mL) = nominal concentration of chloramphenicol CU palmitate in the Sample solution (mg/mL) P = chloramphenicol palmitate equivalent of USP Chloramphenicol Palmitate RS (g/mg) Acceptance criteria: 555595 g SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 8795 SPECIFIC ROTATION 781S: +21 to +25 Sample solution: 50 mg/mL, undried, in dehydrated alcohol CRYSTALLINITY 695: Meets the requirements ACIDITY: Dissolve 1.0 g by heating at 35 with 5 mL of 80% alcohol and ether (1:1), previously neutralized using phenolphthalein TS. Titrate with 0.1 N sodium hydroxide VS, using phenolphthalein TS, until on gentle shaking a pink color persists for NLT 30 s. Acceptance criteria: NMT 0.4 mL is consumed. LOSS ON DRYING 731: Dry it to constant weight over phosphorus pentoxide in vacuum at a pressure not exceeding 5 mm of mercury. Acceptance criteria: It loses NMT 0.5% of its weight. FREE CHLORAMPHENICOL Sample solution: Dissolve 1.0 g in 80 mL of xylene with the aid of gentle warming. Cool, and extract with three 15mL portions of water, combining the aqueous extracts and discarding the xylene. Dilute the combined aqueous extracts with water to 50 mL, extract with 10 mL of toluene, allow to separate, and discard the toluene. Centrifuge a portion of the aqueous solution. Use the clear solution. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 278 nm Analysis: Determine the absorbance of the Sample solution, using as a reagent blank to set the instrument to zero a solution prepared following the procedure for the Sample solution, but omitting the specimen. Acceptance criteria: The absorbance is NMT 0.268 (0.045%). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Chloramphenicol Palmitate RS rU
Chloramphenicol Palmitate
(Comment on this Monograph)id=m15330=Chloramphenicol Palmitate=Chl-Cy-Monos.pdf) 561.54 C27H42Cl2N2O6 Hexadecanoic acid, 2-[(2,2-dichloroacetyl)amino]-3-hydroxy-3(4-nitrophenyl)propyl ester, [R-(R*,R*)]-; D-threo-()-2,2-Dichloro-N-[-hydroxy--(hydroxymethyl)-pnitrophenethyl]acetamide -palmitate [530-43-8]. DEFINITION Chloramphenicol Palmitate has a potency equivalent to NLT 555 g and NMT 595 g of chloramphenicol (C11H12Cl2N2O5)/ mg. IDENTIFICATION The retention time of the chloramphenicol palmitate peak of the Sample solution corresponds to that of the chloramphenicol palmitate peak of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (172:1:27) Standard stock solution: Transfer 65 mg of USP Chloramphenicol Palmitate RS to a 50-mL volumetric flask, add 40 mL of methanol and 1 mL of glacial acetic acid, and sonicate for a few min. Dilute with methanol to volume. Standard solution: 10.0 mL of Standard stock solution diluted with Mobile phase to 25.0 mL Sample stock solution: Transfer 65 mg of Chloramphenicol Palmitate to a 50-mL volumetric flask, add 40 mL of methanol and 1 mL of glacial acetic acid, and sonicate for a few min. Dilute with methanol to volume. Sample solution: 10.0 mL of Sample stock solution diluted with Mobile phase to 25.0 mL Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2400 theoretical plates for the analyte peak
Chloramphenicol Palmitate Oral Suspension

(Comment on this Monograph)id=m15340=Chloramphenicol Palmitate Oral Suspension=Chl-Cy-Monos.pdf) DEFINITION Chloramphenicol Palmitate Oral Suspension contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of chloramphenicol (C11H12Cl2N2O5). It contains one or
USP 32
NLT 14 h. Prepare a 1-in-3 mineral oil dispersion of the dried residue, and place a portion of it between two sodium chloride plates, taking care not to allow air bubbles to form. Analysis: Concomitantly record the absorption spectra of the Standard solution and the Sample solution from 1113 m, with a suitable IR absorption spectrophotometer, using an empty cell to set the instrument to 100% transmittance. Adjust the cell thickness of the Standard solution and the Sample solution so that transmittances of 20%30% are obtained at 12.3 m. On each spectrum, draw a straight baseline between the absorption minima at wavelengths of 11.3 and 12.65 m. Draw straight lines, perpendicular to the wavelength scale, at the wavelengths of maximum absorbance at 11.65 and 11.86 m, intersecting both the baseline and the spectrum. Determine the absorbance ratio: Result = (A11.65a A11.65b)/(A11.86a A11.86b) [NOTEIn the above equation the parenthetic expressions are the differences in absorbance values obtained at the wavelengths indicated by the subscripts for the spectrum (a), and the point of intersection of the perpendicular line with the baseline (b).] Acceptance criteria: The absorbance ratio of the Sample solution is greater than that of the Standard solution, corresponding to NMT 10% of polymorph A. PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For suspension packaged in single-unit containers, it meets the requirements. DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 4.57.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chloramphenicol Palmitate RS USP Chloramphenicol Palmitate Nonpolymorph A RS USP Chloramphenicol Palmitate Polymorph A RS
more suitable buffers, colors, flavors, preservatives, and suspending agents. IDENTIFICATION The retention time of the chloramphenicol palmitate peak of the Sample solution corresponds to that of the chloramphenicol palmitate peak of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (172:1:27) Standard stock solution: 65 mg of USP Chloramphenicol Palmitate RS To a 50-mL volumetric flask, add 40 mL of methanol and 1 mL of glacial acetic acid, and sonicate for a few min. Dilute with methanol to volume. Standard solution: 10.0 mL of Standard stock solution diluted with Mobile phase to 25.0 mL Sample stock solution: Transfer 160 mg of chloramphenicol palmitate from Oral Suspension, wellshaken and free from air bubbles, to a 200-mL volumetric flask containing 20 mL of methanol. Add 4 mL of glacial acetic acid, and dilute with methanol to volume. Sample solution: Pass 20 mL of Sample stock solution through glass-fiber filter paper. Dilute 10.0 mL of filtrate with Mobile phase to 25.0 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2400 theoretical plates from the analyte peak Relative standard deviation: NMT 0.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C11H12Cl2N2O5 equivalent in each mL of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chloramphenicol Palmitate RS in the Standard solution (mg/mL) = concentration of chloramphenicol palmitate in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% rU rS CS OTHER COMPONENTS LIMIT OF POLYMORPH A Standard solution: A dry mixture of 1 part by weight of USP Chloramphenicol Palmitate Polymorph A RS and 9 parts by weight of USP Chloramphenicol Palmitate Nonpolymorph A RS. Prepare a 1-in-3 mineral oil dispersion of this mixture, and place a portion of it between two sodium chloride plates, taking care not to allow air bubbles to form. Sample solution: 20 mL of previously mixed Oral Suspension in a 50-mL centrifuge tube. Add 20 mL of water, and mix. Centrifuge, and discard the supernatant. Add 20 mL of water to the residue in the centrifuge tube, mix, centrifuge, and discard the supernatant. Repeat this washing two times. Dry the residue in a vacuum over silica gel for
Chloramphenicol Sodium Succinate

(Comment on this Monograph)id=m15347=Chloramphenicol Sodium Succinate=Chl-Cy-Monos.pdf) 445.18 C15H15Cl2N2NaO8 Butanedioic acid, mono[2-[(2,2-dichloroacetyl)amino]-3hydroxy-3-(4-nitrophenyl)propyl] ester, monosodium salt, [R(R*,R*)]-; D-threo-()-2,2-Dichloro-N-[-hydroxy--(hydroxymethyl)-pnitrophenethyl]acetamide -(sodium succinate) [982-57-0]. DEFINITION Chloramphenicol Sodium Succinate has a potency equivalent to NLT 650 g and NMT 765 g of chloramphenicol (C11H12Cl2N2O5)/mg. IDENTIFICATION The Sample solution exhibits an absorption maximum at a wavelength of 276 nm, as obtained in the Assay. ASSAY PROCEDURE Standard solution: 20 g/mL of USP Chloramphenicol RS Sample solution: Equivalent to 20 g/mL of chloramphenicol from Chloramphenicol Sodium Succinate
USP 32
Spectrometric conditions Mode: Spectrophotometry Cell: 1 cm Blank: Water Analysis: Concomitantly determine the absorbance of the Standard solution at the wavelength of maximum absorbance at 278 nm, and the absorbance of the Sample solution at the wavelength of maximum absorbance at 276 nm. Calculate the quantity, in g, of C11H12Cl2N2O5 in each mg of Chloramphenicol Sodium Succinate taken: Result = (AU/AS) (CS/CU) P = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) CU = nominal concentration of chloramphenicol in the Sample solution (g/mL) P = potency of USP Chloramphenicol RS (g/mL) Acceptance criteria: 650765 g/mg IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE CHLORAMPHENICOL Solution A: 0.05 M monobasic ammonium phosphate, adjusted with 10% phosphoric acid to a pH of 2.5 0.1 Mobile phase: Methanol and Solution A (2:3) Standard solution: 6 g/mL of USP Chloramphenicol RS in Mobile phase. Pass through a filter having 0.5-m or finer porosity, and use the filtrate. Sample solution: 660 g/mL of Chloramphenicol Sodium Succinate in Mobile phase. Pass a portion through a filter having 0.5-m or finer porosity, and use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard solution and Sample solution Suitability requirements Column efficiency: NLT 1750 theoretical plates from the chloramphenicol-1-succinate and chloramphenicol-3succinate peaks, Sample solution Resolution: NLT 2.0 between the two peaks, Sample solution Tailing factor: NMT 1.2, Sample solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of free C11H12Cl2N2O5 in the portion of Chloramphenicol Sodium Succinate taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of Chloramphenicol Sodium Succinate in the Sample solution (g/mL) AU AS CS

SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +5.0 to +8.0 Sample solution: 50 mg/mL PH 791: 6.47.0, in equivalent to 250 mg/mL of chloramphenicol solution WATER DETERMINATION, Method I 921: NMT 5.0% STERILITY TESTS 71: Where the label states that Chloramphenicol Sodium Succinate is sterile, it meets the requirements. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Chloramphenicol Sodium Succinate must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.2 USP Endotoxin Unit/mg of chloramphenicol. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing sterile dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of sterile dosage forms. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Endotoxin RS
Chloramphenicol Sodium Succinate for Injection

(Comment on this Monograph)id=m15349=Chloramphenicol Sodium Succinate for Injection=Chl-Cy-Monos.pdf) DEFINITION Chloramphenicol Sodium Succinate for Injection contains an amount of Chloramphenicol Sodium Succinate equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of chloramphenicol (C11H12Cl2N2O5). IDENTIFICATION The Sample solution exhibits an absorption maximum at a wavelength of 276 nm, as obtained in the Assay. ASSAY PROCEDURE Standard solution: 20 g/mL of USP Chloramphenicol RS Sample solution: Constitute one container of Injection as directed in the labeling. Prepare a solution equivalent to 20 g/mL of chloramphenicol from Injection in water. Spectrometric conditions Mode: Spectrophotometry Cell: 1 cm Blank: Water Analysis: Concomitantly determine the absorbance of the Standard solution, at the wavelength of maximum absorbance at 278 nm, and the absorbance of the Sample solution, at the wavelength of maximum absorbance at 276 nm. Calculate the percentage of C11H12Cl2N2O5 in each mL of the constituted portion of Injection taken: Result = (AU/AS) (CS/CU) 100 AU AS = absorbance of the Sample solution = absorbance of the Standard solution

= concentration of USP Chloramphenicol RS in the Standard solution (g/mL) = nominal concentration of chloramphenicol in the CU Sample solution (g/mL) Acceptance criteria: 90.0%115.0% CS WATER DETERMINATION, Method I 921: NMT 5.0%
USP 32
IMPURITIES Organic Impurities PROCEDURE: LIMIT OF FREE CHLORAMPHENICOL Solution A: 0.05 M monobasic ammonium phosphate, adjust the pH with 10% phosphoric acid to 2.5 0.1 Mobile phase: Methanol and Solution A (2:3) Standard solution: 6 g/mL of USP Chloramphenicol RS in Mobile phase, pass through a 0.5-m or finer porosity filter, and use the filtrate Sample stock solution: Equivalent of 100 mg/mL of chloramphenicol by dissolving the contents of one container in a volume of Mobile phase Sample solution: Equivalent of 0.5 mg/mL of chloramphenicol from Sample stock solution in Mobile phase. Pass through a 0.5-m or finer porosity filter, and use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 275 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard solution and Sample solution Suitability requirements Column efficiency: NLT 1750 theoretical plates from chloramphenicol-1-succinate and chloramphenicol-3succinate peaks, Sample solution Resolution: NLT 2.0 between the two peaks, Sample solution Tailing factor: NMT 1.2, Sample solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of free C11H12Cl2N2O5 in the sample taken: Result = (rU/rS) (CS/CU) 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Chloramphenicol RS in the Standard solution (g/mL) CU = nominal concentration of chloramphenicol in the Sample solution (g/mL) Acceptance criteria: NMT 2.0% rU rS CS SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.2 USP Endotoxin Unit/mg of chloramphenicol. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections. OPTICAL ROTATION, Specific Rotation 781S: +5.0 to +8.0 Sample solution: 50 mg/mL PH 791: 6.47.0, in equivalent to 250 mg/mL of chloramphenicol solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Chloramphenicol RS USP Endotoxin RS
Chlordiazepoxide
(Comment on this Monograph)id=m15450=Chlordiazepoxide=Chl-Cy-Monos.pdf)
299.75 C16H14ClN3O 3H-1,4-Benzodiazepin-2-amine, 7-chloro-N-methyl-5-phenyl-, 4oxide; 7-Chloro-2-(methylamino)-5-phenyl-3H-1,4-benzodiazepine 4oxide [58-25-3]. DEFINITION Chlordiazepoxide contains NLT 98.0% and NMT 102.0% of C16H14ClN3O, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. PROCEDURE Sample solution: To 20 mg, add 5 mL of hydrochloric acid and 10 mL of water, and heat to boiling to effect hydrolysis. Analysis: To the cooled Sample solution, add 2 mL of sodium nitrite solution (1 in 1000), shake, add 1 mL of ammonium sulfamate solution (1 in 200), then shake for 2 min, and add 1 mL of N-(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: A reddish violet color is produced. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: Methanol and water (3:2) Standard solution: 200 g/mL of USP Chlordiazepoxide RS in Mobile phase Sample stock solution: 100 mg of Chlordiazepoxide to a 50-mL volumetric flask, dissolve in Mobile phase, sonicate for 5 min, dilute with Mobile phase to volume, and mix. Pass this solution through a membrane filter having a 0.5-m or finer porosity. Sample solution: 10 mL of the Sample stock solution to a 100-mL volumetric flask, and dilute with Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3600 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H14ClN3O in the portion of Chlordiazepoxide taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlordiazepoxide RS in the Standard solution (g/mL) = concentration of Chlordiazepoxide in the Sample CU solution (g/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Standard solution A: 100 g/mL of USP Chlordiazepoxide Related Compound A RS in acetone Standard solution B: 10 g/mL of USP 2-Amino-5chlorobenzophenone RS in acetone Sample solution: 50.0 mg to a 10-mL conical flask, add 2.5 mL of acetone, and shake. Allow any undissolved particles to settle, and use the supernatant. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 50 L for Sample solution, and 10 L for Standard solution A and Standard solution B Developing solvent system: Ethyl acetate Spray reagent: 2 N sulfuric acid Analysis Samples: Standard solutions and Sample solution Develop the chromatogram in a chromatographic chamber not previously saturated with the Developing solvent system. Locate the spots on the plate by lightly spraying with Spray reagent, drying at 105 for 15 min, and then spraying in succession with sodium nitrite solution (1 in 1000), ammonium sulfamate solution (1 in 200), and N(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots at the respective RF values produced by the Standard solutions, corresponding to NMT 0.1% of chlordiazepoxide related compound A, and NMT 0.01% of 2-amino-5chlorobenzophenone. SPECIFIC TESTS LOSS ON DRYING 731: 0.3% of its weight. Dry at 105 for 3 h: it loses NMT
Official Monographs / Chlordiazepoxide 9

USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Chlordiazepoxide RS USP Chlordiazepoxide Related Compound A RS
Chlordiazepoxide Tablets
(Comment on this Monograph)id=m15480=Chlordiazepoxide Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlordiazepoxide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C16H14ClN3O. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. PROCEDURE Sample solution: 20 mg from finely powdered Tablets, add 5 mL of hydrochloric acid and 10 mL of water, and heat to boiling to effect hydrolysis Analysis: To the cooled Sample solution, add 2 mL of sodium nitrite solution (1 in 1000), shake, add 1 mL of ammonium sulfamate solution (1 in 200), then shake for 2 min, and add 1 mL of N-(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: A reddish violet color is produced. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: Methanol and water (3:2) Standard solution: 200 g/mL of USP Chlordiazepoxide RS in Mobile phase Sample solution: 5 mg of chlordiazepoxide from finely powdered Tablets (NLT 20), to a 25-mL volumetric flask, add 20 mL of Mobile phase, sonicate for 5 min to dissolve, dilute with Mobile phase to volume, and pass through a 5m membrane filter, discarding the first 5 mL of the filtrate Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3600 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H14ClN3O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlordiazepoxide RS in the Standard solution (g/mL) = nominal concentration of chlordiazepoxide in the Sample solution (g/mL)
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Chlordiazepoxide / Official Monographs

90.0%110.0
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IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Solution A: 2.402 mg/mL of glacial acetic acid, 4.612 mg/mL of phosphoric acid, and 2.473 mg/mL of boric acid, in water Solution B: 0.20 N sodium hydroxide solution and Solution A (10.5:100) Solution C: Methanol, tetrahydrofuran, and Solution B (1:4:5) Mobile phase: 0.01 M sodium lauryl sulfate in Solution C Standard stock solution: 1 mg/mL of USP Chlordiazepoxide RS and 2.8 mg/mL of USP Amitriptyline Hydrochloride RS in Solution C Standard solution: 1.0 mL of Standard stock solution diluted with Solution C to 10.0 mL Sample stock solution: 50 mg of chlordiazepoxide and 125 mg of amitriptyline from finely powdered Tablets (NLT 20), to a 50-mL volumetric flask, add Solution C to volume, sonicate to disperse the mixture, and allow undissolved particles to settle Sample solution: 1.0 mL of Sample stock solution diluted with Solution C to 10.0 mL. Pass a portion of this solution through a 0.5-m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe retention times for chlordiazepoxide and amitriptyline hydrochloride are about 5 min and 7 min, respectively.] Suitability requirements Resolution: NLT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H14ClN3O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlordiazepoxide RS in the Standard solution (g/mL) CU = nominal concentration of chlordiazepoxide in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% Calculate the percentage of C20H23N in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 Mr2 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Amitriptyline Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of amitriptyline hydrochloride in the Sample solution (g/mL) = molecular weight of amitriptyline, 277.40 = molecular weight of amitriptyline hydrochloride, 313.86 rU rS CS
PERFORMANCE TESTS DISSOLUTION 711 Medium: Simulated gastric fluid TS, prepared without pepsin; 900 mL Apparatus 1: 100 rpm Time: 30 min Detector: UV 309 nm Standard solution: USP Chlordiazepoxide RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Tolerances: NLT 85% (Q) of the labeled amount of C16H14ClN3O is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution A: 1 mg/mL of USP Chlordiazepoxide Related Compound A RS in acetone Standard solution B: 100 g/mL of USP 2-Amino-5chlorobenzophenone RS in acetone Sample solution: Transfer the equivalent of 25 mg of chlordiazepoxide from finely powdered Tablets to a 10-mL conical flask, add 2.5 mL of acetone, and shake. Allow any undissolved particles to settle, and use the supernatant. Spray reagent: 2 N sulfuric acid Application volume: 50 L for Sample solution, 20 L for Standard solution A, and 5 L for Standard solution B Developing solvent system: Ethyl acetate Analysis Samples: Standard solutions A and B, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in a chromatographic chamber (not previously saturated with the Developing solvent system). Locate the spots on the plate by lightly spraying with Spray reagent, drying at 105 for 15 min, and then spraying in succession with sodium nitrite solution (1 in 1000), ammonium sulfamate solution (1 in 200), and N-(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots at the respective RF values produced by the Standard solutions, corresponding to NMT 4.0% of chlordiazepoxide related compound A, and to NMT 0.1% of 2-amino-5chlorobenzophenone. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Chlordiazepoxide RS USP Chlordiazepoxide Related Compound A RS
Chlordiazepoxide and Amitriptyline Hydrochloride Tablets

(Comment on this Monograph)id=m15510=Chlordiazepoxide and Amitriptyline Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlordiazepoxide and Amitriptyline Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of chlordiazepoxide (C16H14ClN3O) and an amount of amitriptyline hydrochloride equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of amitriptyline (C20H23N).
USP 32

UNIFORMITY OF DOSAGE UNITS, Content Uniformity 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Standard solution A: 1 mg/mL of USP Chlordiazepoxide Related Compound A RS in acetone Standard solution B: 50 g/mL of USP 2-Amino-5chlorobenzophenone RS in acetone Sample solution: 25 mg of chlordiazepoxide from finely powdered Tablets, to a 10-mL conical flask, add 2.5 mL of acetone, and shake. Allow any undissolved particles to settle, and use the supernatant. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 50 L for Sample solution, 20 L for Standard solution A, and 10 L for Standard solution B Developing solvent system: Ethyl acetate Spray reagent: 2 N sulfuric acid Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed in the chapter. Develop the chromatogram in a chromatographic chamber (not previously saturated with the Developing solvent system). Locate the spots on the plate by lightly spraying with Spray reagent, drying at 105 for 15 min, and then spraying in succession with sodium nitrite solution (1 in 1000), ammonium sulfamate solution (1 in 200), and N(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots at the respective RF values produced by the Standard solutions, corresponding to NMT 4.0% of chlordiazepoxide related compound A, and NMT 0.1% of 2-amino-5chlorobenzophenone. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Amitriptyline Hydrochloride RS USP Chlordiazepoxide RS USP Chlordiazepoxide Related Compound A RS
PERFORMANCE TESTS DISSOLUTION 711 Medium: Simulated gastric fluid TS, prepared without pepsin; 900 mL Apparatus 1: 100 rpm Time: 30 min Detector: UV 239 nm and 309 nm Blank: Medium Standard solution A: USP Chlordiazepoxide RS in Medium Standard solution B: USP Amitriptyline Hydrochloride RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Analysis: Calculate the percentage of C16H14ClN3O dissolved: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution measured at 309 nm = absorbance of Standard solution A measured at AS 309 nm = concentration of USP Chlordiazepoxide RS in CS Standard solution A measured at 309 nm (mg/mL) = nominal concentration of chlordiazepoxide in the CU Sample solution (mg/mL) Calculate the percentage of C20H23N dissolved: AU Result = (AX/AS) (CS/CU) (Mr1/Mr2) 100 = absorbance variable calculated as shown in the formula below = absorbance of Standard solution B measured at AS 239 nm CS = concentration of USP Amitriptyline Hydrochloride RS in Standard solution B measured at 239 nm (mg/mL) = nominal concentration of amitriptyline CU hydrochloride in the Sample solution (mg/mL) = molecular weight of amitriptyline, 277.40 Mr1 = molecular weight of amitriptyline hydrochloride, Mr2 313.86 The absorbance value AX in the formula above is calculated as follows: Result = (AU239 AU309) [(C309 AS239)/(C239 AS309)] = absorbance of the Sample solution measured at 239 nm AU309 = absorbance of the Sample solution measured at 309 nm = concentration of USP Chlordiazepoxide RS in C309 Standard solution A measured at 309 nm (mg/mL) AS239 = absorbance of Standard solution A measured at 239 nm = concentration of USP Chlordiazepoxide RS in C239 Standard solution A measured at 239 nm (mg/mL) AS309 = absorbance of Standard solution A measured at 309 nm [NOTEAll of the chlordiazepoxide measurements may be made with either a single Standard solution, or two separate Standard solutions.] Tolerances: NLT 85% (Q) of the labeled amount of C16H14ClN3O, and an amount of amitriptyline hydrochloride equivalent to NLT 85% (Q) of the labeled amount of C20H23N is dissolved. AU239 AX
Chlordiazepoxide Hydrochloride
(Comment on this Monograph)id=m15520=Chlordiazepoxide Hydrochloride=Chl-Cy-Monos.pdf) 336.22 C16H14ClN3O HCl 3H-1,4-Benzodiazepin-2-amine, 7-chloro-N-methyl-5-phenyl-, 4oxide, monohydrochloride; 7-Chloro-2-(methylamino)-5-phenyl-3H-1,4-benzodiazepine 4oxide monohydrochloride [438-41-5]. DEFINITION Chlordiazepoxide Hydrochloride contains NLT 98.0% and NMT 102.0% of C16H14ClN3O HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The relative retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
12
USP 32
Developing solvent system: Ethyl acetate Spray reagent: 2 N sulfuric acid Analysis Samples: Standard solutions and Sample solution Proceed as directed in the chapter. Develop the chromatogram in a chromatographic chamber (not previously saturated with the Developing solvent system). Locate the spots on the plate by lightly spraying with Spray reagent, drying at 105 for 15 min, and then spraying in succession with sodium nitrite solution (1 in 1000), ammonium sulfamate solution (1 in 200), and N(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots at the respective RF values produced by the Standard solutions, corresponding to NMT 0.1% of chlordiazepoxide related compound A, and to NMT 0.01% of 2-amino-5chlorobenzophenone. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 212218, with decomposition LOSS ON DRYING 731: Dry it in a vacuum over phosphorus pentoxide at 60 for 4 h: it loses NMT 0.5% of its weight. STERILITY TESTS 71 (where the label states that Chlordiazepoxide Hydrochloride is sterile): Meets the requirements BACTERIAL ENDOTOXINS TEST 85: It contains NMT 3.57 USP Endotoxin Units/mg of chlordiazepoxide hydrochloride. OTHER REQUIREMENTS: It meets the requirements for Labeling under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Chlordiazepoxide Hydrochloride RS USP Chlordiazepoxide Related Compound A RS USP Endotoxin RS
C. PROCEDURE Sample solution: To 20 mg, add 5 mL of hydrochloric acid and 10 mL of water, and heat to boiling to effect hydrolysis. Analysis: To the cooled Sample solution, add 2 mL of sodium nitrite solution (1 in 1000), 1 mL of ammonium sulfamate solution (1 in 200), and 1 mL of N-(1naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: A reddish-violet color is produced. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: Methanol and water (3:2) Standard solution: 200 g/mL of USP Chlordiazepoxide Hydrochloride RS in Mobile phase Sample stock solution: 2 mg/mL of Chlordiazepoxide Hydrochloride in Mobile phase [NOTESonicate if necessary.] Sample solution: 0.2 mg/mL from Sample stock solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3600 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H14ClN3O HCl in the portion of Chlordiazepoxide Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlordiazepoxide Hydrochloride RS in the Standard solution (g/mL) = concentration of chlordiazepoxide hydrochloride CU in the Sample solution (g/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Standard solution A: 100 g/mL of USP Chlordiazepoxide Related Compound A RS in acetone Standard solution B: 10 g/mL of USP 2-Amino-5chlorobenzophenone RS in acetone Sample solution: 50.0 mg to a 10-mL conical flask, add 2.5 mL of acetone, and shake. Allow any undissolved particles to settle, and use the supernatant. Chromatographic System (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 50 L for the Sample solution and 10 L for the Standard solutions rU rS CS
Chlordiazepoxide Hydrochloride Capsules

(Comment on this Monograph)id=m15530=Chlordiazepoxide Hydrochloride Capsules=Chl-Cy-Monos.pdf) DEFINITION Chlordiazepoxide Hydrochloride Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of C16H14ClN3O HCl. IDENTIFICATION A. The Sample solution as prepared in the Assay exhibits maxima at 245 2 nm and 311 2 nm, and the ratio A245/A311 is between 2.90 and 3.45. B. PROCEDURE Sample solution: To the equivalent of 20 mg of chlordiazepoxide hydrochloride from Capsule contents, add 5 mL of hydrochloric acid and 10 mL of water, and heat to boiling to effect hydrolysis.
USP 32
Analysis: To the cooled Sample solution add 2 mL of sodium nitrite solution (1 in 1000), 1 mL of ammonium sulfamate solution (1 in 200), and 1 mL of N-(1naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: A reddish-violet color is produced. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: Methanol and water (3:2) Standard solution: 200 g/mL of USP Chlordiazepoxide Hydrochloride RS in Mobile phase Sample solution: 0.2 mg/mL of chlordiazepoxide hydrochloride from Capsule contents, in Mobile phase [NOTESonicate if necessary.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3600 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H14ClN3O in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlordiazepoxide Hydrochloride RS in the Standard solution (g/mL) = concentration of chlordiazepoxide hydrochloride CU in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 30 min Detector: UV 245 nm, in 1-cm cells Sample solution: Sample per Dissolution 711. Remove the contents of 12 Capsules as completely as possible with the aid of a current of air. Dissolve the empty capsule shells in 900 mL of Medium. Filter a portion of the solution. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Chlordiazepoxide Hydrochloride RS in Medium Tolerances: NLT 85% (Q) of the labeled amount of C16H14ClN3O HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity [NOTEUse low-actinic glassware.] Sample stock solution: Transfer the contents of 1 Capsule to a 200-mL volumetric flask, dissolve in and dilute with water to volume, and filter, discarding the first 20 mL of the filtrate. Sample solution: 6 g/mL of chlordiazepoxide hydrochloride in 0.1 N hydrochloric acid, from Sample stock solution rU rS CS

Standard solution: 6 g/mL of USP Chlordiazepoxide Hydrochloride RS in 0.1 N hydrochloric acid Spectroscopic conditions Mode: Spectrophotometry Analytical wavelength: 245 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H14ClN3O HCl in the Capsule taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Chlordiazepoxide Hydrochloride RS in the Standard solution (g/mL) = concentration of chlordiazepoxide hydrochloride in the Sample solution (g/mL)
IMPURITIES Organic Impurities PROCEDURE Standard solution A: 1 mg/mL of USP Chlordiazepoxide Related Compound A RS in acetone Standard solution B: 50 g/mL of USP 2-Amino-5chlorobenzophenone RS in acetone Sample solution: 25 mg of chlordiazepoxide hydrochloride from Capsule contents, to a 10-mL conical flask. Add 2.5 mL of acetone, and shake. Allow any undissolved particles to settle, and use the supernatant. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 50 L for Sample solution, 15 L for Standard solution A, and 10 L for Standard solution B Developing solvent system: Ethyl acetate Spray reagent: 2 N sulfuric acid Analysis Samples: Standard solutions and Sample solution Proceed as directed in the chapter. Develop the chromatogram in a chromatographic chamber (not previously saturated with the Developing solvent system). Locate the spots on the plate by lightly spraying with Spray reagent, drying at 105 for 15 min, and then spraying in succession with sodium nitrite solution (1 in 1000), ammonium sulfamate solution (1 in 200), and N(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots at the respective RF values produced by the Standard solutions, corresponding to NMT 3.0% of chlordiazepoxide related compound A, and to NMT 0.1% of 2-amino-5chlorobenzophenone. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Chlordiazepoxide Hydrochloride RS USP Chlordiazepoxide Related Compound A RS
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Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 50 L for the Sample solution and 10 L for Standard solutions A and B Developing solvent system: Ethyl acetate Spray reagent: 2 N sulfuric acid Analysis Samples: Standard solutions A and B and Sample solution Develop the chromatogram in a chromatographic chamber (not previously saturated with the developing solvent) in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by lightly spraying with Spray reagent, drying at 105 for 15 min, and then spraying in succession with sodium nitrite solution (1 in 1000), ammonium sulfamate solution (1 in 200), and N(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots at the respective RF values produced by Standard solutions A and B, corresponding to NMT 0.1% of chlordiazepoxide related compound A, and to NMT 0.01% of 2-amino-5chlorobenzophenone. SPECIFIC TESTS PH 791: 2.53.5, in a solution (1 in 100) COMPLETENESS OF SOLUTION 641: It dissolves in the solvent and in the concentration recommended in the labeling to yield a clear solution. CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Labels and Labeling and Constituted Solutions under Injections 1. LOSS ON DRYING 731: Dry in a vacuum over phosphorus pentoxide at 60 for 4 h: it loses NMT 0.5% of its weight. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 3.57 USP Endotoxin Units/mg of chlordiazepoxide hydrochloride STERILITY TESTS 71: Meets the requirements OTHER REQUIREMENTS: It meets the requirements for Labels and Labeling under Injections 1. PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Chlordiazepoxide Hydrochloride for Injection

(Comment on this Monograph)id=m15535=Chlordiazepoxide Hydrochloride for Injection=Chl-Cy-Monos.pdf) DEFINITION Chlordiazepoxide Hydrochloride for Injection is Chlordiazepoxide Hydrochloride suitable for parenteral use. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The relative retention time of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. To 20 mg add 5 mL of hydrochloric acid and 10 mL of water, and heat to boiling to effect hydrolysis. To the cooled solution add 2 mL of sodium nitrite solution (1 in 1000), 1 mL of ammonium sulfamate solution (1 in 200), and 1 mL of N-(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000): a reddish violet color is produced. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Mobile phase: Methanol and water (3:2) Standard solution: 200 g/mL of USP Chlordiazepoxide Hydrochloride RS in Mobile phase Sample stock solution: 2 mg/mL of Chlordiazepoxide Hydrochloride for Injection in Mobile phase Sample solution: 0.2 mg/mL from Sample stock solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3600 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution. Calculate the percentage of chlordiazepoxide hydrochloride in the portion of Chlordiazepoxide Hydrochloride for Injection taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlordiazepoxide Hydrochloride RS in the Standard solution (g/mL) = concentration of chlordiazepoxide hydrochloride in the Sample solution (g/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids, as described under Injections 1, protected from light. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Chlordiazepoxide Hydrochloride RS USP Chlordiazepoxide Related Compound A RS USP Endotoxin RS
IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Standard solution A: 100 g/mL of USP Chlordiazepoxide Related Compound A RS in acetone Standard solution B: 10 g/mL of USP 2-Amino-5chlorobenzophenone RS in acetone Sample solution: Transfer 50.0 mg to a 10-mL conical flask, add 2.5 mL of acetone, and shake. Allow any undissolved particles to settle, and use the supernatant.
Chlordiazepoxide Hydrochloride and Clidinium Bromide Capsules

(Comment on this Monograph)id=m15560=Chlordiazepoxide Hydrochloride and Clidinium Bromide Capsules=Chl-CyMonos.pdf) DEFINITION Chlordiazepoxide Hydrochloride and Clidinium Bromide Capsules contain NLT 90.0% and NMT 110.0% of the labeled amounts of chlordiazepoxide hydrochloride (C16H14ClN3O HCl) and clidinium bromide (C22H26BrNO3).
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IDENTIFICATION The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEUse low-actinic glassware.] Solution A: 1.92 mg/mL of sodium 1-pentanesulfonate in water. Adjust with 1 N sulfuric acid to a pH of 3.8 0.1. Mobile phase: Methanol, tetrahydrofuran, and Solution A (3:12:35) Diluent: Methanol and water (1:1) Standard solution: 0.1 mg/mL of USP Chlordiazepoxide Hydrochloride RS and 0.05 mg/mL of USP Clidinium Bromide RS in Diluent Sample solution: Weigh the contents of NLT 20 Capsules, and calculate the average weight per Capsule. Mix the combined contents of the Capsules, and transfer an equivalent to about 5 mg of C16H14ClN3O HCl to a 50-mL volumetric flask. Add about 25 mL of Diluent, sonicate for 5 min, and shake by mechanical means for 10 min. Dilute with Diluent to volume and filter, discarding the first 20 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 212 nm Column: 8 mm 10 cm; packing L1 Flow rate: 3 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for clidinium bromide and chlordiazepoxide hydrochloride are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 5.0 between the clidinium bromide and chlordiazepoxide hydrochloride peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H14ClN3O HCl in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = chlordiazepoxide hydrochloride peak response from the Sample solution rS = chlordiazepoxide hydrochloride peak response from the Standard solution = concentration of USP Chlordiazepoxide CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of chlordiazepoxide CU hydrochloride in the Sample solution (mg/mL) Calculate the percentage of C22H26BrNO3 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = clidinium bromide peak response from the Sample solution = clidinium bromide peak response from the Standard solution = concentration of USP Clidinium Bromide RS in the Standard solution (mg/mL) = nominal concentration of clidinium bromide in the Sample solution (mg/mL) rU

PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 30 min Determine the amounts of chlordiazepoxide hydrochloride and clidinium bromide dissolved using the following method. Solution A: 1.92 mg/mL of sodium 1-pentanesulfonate in water. Adjust with dilute sulfuric acid (1 in 100) to a pH of 3.8 0.1. Mobile phase: Methanol, tetrahydrofuran, and Solution A (6:18:75) Standard solution: Separately prepare USP Chlordiazepoxide Hydrochloride RS and USP Clidinium Bromide RS in Medium. Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 212 nm Column: 4 mm 25 cm; packing L1 Flow rate: 2 mL/min Injection size: 100 L System suitability Sample: Standard solution [NOTEThe relative retention times for clidinium bromide and chlordiazepoxide hydrochloride are about 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 5.0 between the two components Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the amounts of C16H14ClN3O HCl and C22H26BrNO3 dissolved in comparison with known concentrations of the Standard solution. Tolerances: NLT 75% (Q) each of the labeled amounts of C16H14ClN3O HCl and C22H26BrNO3 are dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF CHLORDIAZEPOXIDE RELATED COMPOUND A AND 2-AMINO-5-CHLOROBENZOPHENONE Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution A: 1 mg/mL of USP Chlordiazepoxide Related Compound A RS in acetone Standard solution B: 50 g/mL of USP 2-Amino-5chlorobenzophenone RS in acetone Sample solution: Transfer 25 mg of chlordiazepoxide hydrochloride from Capsule contents, to a 10-mL conical flask, add 2.5 mL of acetone, and shake. Allow any undissolved particles to settle, and use the supernatant. Spray reagent: 2 N sulfuric acid Application volume: 50 L for the Sample solution, 15 L for Standard solution A, and 10 L for Standard solution B Developing solvent system: Ethyl acetate Analysis Samples: Standard solutions and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in a chromatographic chamber (not previously saturated with the developing solvent) in the Developing solvent system
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Extracting solvent mixture: Dehydrated alcohol and cyclohexane (1:1) Standard solution: 50 mg/mL of USP Clidinium Bromide RS in 0.1 N methanolic hydrochloric acid [NOTEPrepare this solution at the time of use.] Identification solution: 50 mg/mL of USP Clidinium Bromide RS in 0.1 N methanolic hydrochloric acid, and to 1 mL of this solution add 20 L of a solution of 25 mg/mL of USP Clidinium Bromide Related Compound A RS in methanol [NOTEPrepare this solution at the time of use.] Sample solution: Empty a number of Capsules, equivalent to 25 mg of clidinium bromide, into a glass-stoppered centrifuge tube, and add 5 mL of the Extracting solvent mixture. Heat the tube gently, with shaking, to 50, centrifuge, and decant the clear supernatant into a second tube. Repeat the addition of Extracting solvent mixture twice, heating, centrifuging, and decanting as before, and combine the three extracts in a single tube. Gently heating, evaporate the combined extracts under a stream of nitrogen to dryness. Dissolve the residue in 0.5 mL of methanol. Chromatographic plates: Predevelop suitable thin-layer chromatographic plates (see Chromatography 621, ThinLayer Chromatography) by placing in a chromatographic chamber saturated with Developing solvent system, and allow the Developing solvent system to move 15 cm. Remove the plates from the chamber, dry at 105 for 15 min, and cool. Solution A: 17 mg/mL of bismuth subnitrate in a mixture of glacial acetic acid and water (1:4) Solution B: 0.4 g/mL of potassium iodide Application volume: 20 L Developing solvent system: Acetone, methanol, water, and hydrochloric acid (14:4:1:1) Spray reagent: Solution A, Solution B, and dilute sulfuric acid (1 in 10) (1:1:8). Add 15 5 g/L of iodine, and mix until solution is complete. Analysis Samples: Standard solution, Identification solution, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Place the plates in an unsaturated chromatographic chamber containing freshly prepared Developing solvent system, and develop the chromatogram until the solvent front has moved 15 cm. Remove the plates, and dry at 105 for 10 min. Cool to room temperature, and spray with Spray reagent. Any spot in the chromatogram of the Sample solution occurring at an RF value of 0.4 is not greater in size or intensity than the corresponding spot in the chromatogram of the Identification solution; and the Standard solution shows no spot at the RF value corresponding to that of clidinium bromide related compound A. Acceptance criteria: NMT 1.0% of clidinium bromide related compound A ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Chlordiazepoxide Hydrochloride RS USP Chlordiazepoxide Related Compound A RS USP Clidinium Bromide RS USP Clidinium Bromide Related Compound A RS USP 3-Quinuclidinyl Benzilate RS
until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by lightly spraying with Spray reagent, drying at 105 for 15 min, and then spraying in succession with sodium nitrite solution (1 in 1000), ammonium sulfamate solution (1 in 200), and N-(1-naphthyl)ethylenediamine dihydrochloride solution (1 in 1000). Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots at the respective RF values produced by the Standard solutions, corresponding to NMT 3.0% of chlordiazepoxide related compound A, and to NMT 0.1% of 2-amino-5chlorobenzophenone. PROCEDURE 2: LIMIT OF 3-QUINUCLINDINYL BENZILATE Adsorbent: 0.25-mm layer of chromatographic silica gel Sample solution: Place a glass wool plug in the bottom of a 2.5-cm 35 5-cm glass chromatographic tube, add 2 g of chromatographic siliceous earth triturated with 1 mL of 1 N hydrochloric acid, and lightly pack with a glass tamping rod. Empty a number of Capsules, equivalent to 15 mg of clidinium bromide, into a 100-mL beaker, add 3 mL of 1 N hydrochloric acid, and swirl to dissolve. Add 4 g of chromatographic siliceous earth, mix with a spatula, and add the Capsulesiliceous earth mixture to the chromatographic tube. Dry-wash the beaker with an additional 0.51.0 g of chromatographic siliceous earth, adding the washing to the top of the column. Lightly pack with the tamping rod, and overlay the column with glass wool. Insert the lower exit tube of the column into a 125mL separator, and elute the column with 100 mL of chloroform previously distilled over 1 N sulfuric acid and saturated with water. Extract the chloroform eluate with 20 mL of freshly prepared ascorbic acid solution (1 in 20), preserving the extract. Extract the eluate with a second 15mL portion of the ascorbic acid solution, combine the extracts in the separator, and discard the chloroform layer. Neutralize the acid extracts by adding sodium bicarbonate until the solution is slightly alkaline to the pH paper. Extract the slightly alkaline solution with two 25-mL portions of chloroform, combine the chloroform extracts, and pass through dry, fluted filter paper into a 100-mL beaker. Evaporate the chloroform to dryness with the aid of a stream of nitrogen, and transfer the residue to a glassstoppered 1.0 mL volumetric flask, using methanol to facilitate the transfer. Dilute with methanol to volume. Identification solution: 30 g/mL of USP 3-Quinuclidinyl Benzilate RS in methanol Spray reagent: Potassium iodoplatinate TS Application volume: 100 L for the Sample solution, 15 L for the Identification solution Developing solvent system: Methanol Analysis Samples: Sample solution and Identification solution Place the plate in a paper-lined, methanol-saturated chromatographic chamber, and proceed as directed for Chromatography 621, Thin-layer Chromatography. Remove the plate, air-dry, spray with Spray reagent, and allow the spots to develop for 10 min. Any spot in the chromatogram of the Sample solution occurring at an RF value of 0.3 is not greater in size or intensity than the spot in the chromatogram of the Identification solution. Acceptance criteria: NMT 0.03% of 3-quinuclidinyl benzilate PROCEDURE 3: LIMIT OF CLIDINIUM BROMIDE RELATED COMPOUND A Adsorbent: 0.25-mm layer of chromatographic silica gel mixture
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Official Monographs / Chlorhexidine 17

Time (min) 16 21 Solution A (%) 100 100 Solution B (%) 0 0
Chlorhexidine Gluconate Oral Rinse

(Comment on this Monograph)id=m15629=Chlorhexidine Gluconate Oral Rinse=Chl-Cy-Monos.pdf) DEFINITION Chlorhexidine Gluconate Oral Rinse is prepared from Chlorhexidine Gluconate Solution. It contains NLT 90.0% and NMT 110.0% of the labeled amount of chlorhexidine gluconate (C22H30Cl2N10 2C6H12O7). IDENTIFICATION A. The retention time of the major peak for chlorhexidine from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. To a volume of Oral Rinse, equivalent to 10 mg of chlorhexidine gluconate, add 5 mL of a solution of cetyltrimethylammonium bromide (1 in 100), 1 mL of 10 N sodium hydroxide, and 1 mL of bromine TS: a deep red color is produced. C. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 0.6 mg/mL of USP Potassium Gluconate RS Sample solution: Undiluted Oral Rinse Chromatographic system (See Chromatography 621, System Suitability.) Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 15 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3) Spray reagent: Dissolve 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask, add 1.0 g of ceric sulfate, swirl to dissolve, and dilute with 2 N sulfuric acid to volume. Analysis Samples: Standard solution and Sample solution Develop the chromatogram until the solvent front has moved 10 cm from the point of spotting. Remove the plate from the chamber, and dry at 110 for 20 min. Allow to cool, and spray with Spray reagent. Heat the plate at 110 for 10 min. Acceptance criteria: The principal spot of the Sample solution corresponds in color, size, and RF value to that of the Standard solution. ASSAY PROCEDURE Diluent: Prepare a solution of 27.6 g of monobasic sodium phosphate in about 1.5 L of water. Adjust with phosphoric acid to a pH of 3.0, and dilute with water to 2000 mL. Solution A: Dissolve 27.6 g of monobasic sodium phosphate and 10 mL of triethylamine in 1.5 L of water. Adjust with phosphoric acid to a pH of 3.0, and dilute with water to 2000 mL. Prepare a mixture of the resulting solution and acetonitrile (70:30). [NOTESmall adjustments in the acetonitrile content may be made to meet acceptable resolution criteria.] Degas before use, and sparge with helium during the analysis. Solution B: Acetonitrile Mobile phase: Use variable mixtures of Solution A and Solution B as directed in the gradient table below.
System suitability solution: 50 g/mL of USP Chlorhexidine Acetate RS and 1 g/mL of p-chloroaniline in Diluent Standard solution: 1 mg/mL of USP Chlorhexidine Acetate RS in water. Dilute with Diluent to obtain a 50 g/mL solution. Sample solution: Nominally 60 g/mL of chlorhexidine gluconate from the Oral Rinse diluted with Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 239 nm Column: 4.6-mm 25-cm; base-deactivated 5-m packing L1 Temperature: 40 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 3.0 between chlorhexidine and pchloroaniline Relative standard deviation: NMT 2.0% from the chlorhexidine peak, NMT 5.0% from the p-chloroaniline peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30Cl2N10 2C6H12O7 in the portion of Oral Rinse taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = chlorhexidine peak area response from the Sample solution = chlorhexidine peak area response from the rS Standard solution = concentration of USP Chlorhexidine Acetate RS CS in the Standard solution (g/mL) nominal concentration of chlorhexidine gluconate CU in the Sample solution (g/mL) = molecular weight of chlorhexidine gluconate, Mr1 897.76 = molecular weight of chlorhexidene acetate, Mr2 625.55 Acceptance criteria: 90.0%110.0% rU OTHER COMPONENTS CONTENT OF ALCOHOL Internal standard solution: Dilute 25 mL of n-propyl alcohol with water to 500 mL. Standard solution: Transfer 0.25 g of dehydrated alcohol to a 28-mL screw-capped vial containing 3 mL of water. Add 5.0 mL of Internal standard solution, and dilute with water to almost fill the vial. Cap the vial, and using a vortex mixer, mix for 15 s. Sample solution: Transfer 2.5 g of Oral Rinse to a 28-mL screw-capped vial. Add 5.0 mL of Internal standard solution, and dilute with water to almost fill the vial. Cap the vial, and using a vortex mixer, mix for 15 s. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.53-mm 30-m column, the internal wall coated with a 1.5-m film of liquid phase G27
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Chlorhexidine / Official Monographs

Acceptance criteria: SPECIFIC TESTS PH 791: 5.07.0 NMT 3 g/mL
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Temperature Column Between periods of use: 150 At the time of use: The initial column temperature is maintained at 35 until the alcohol peaks elute, then is increased at a rate of 30/min to a final temperature of 225. Detector: 275 Injector: 250 Injection type: Split injection port Split ratio: 10:1 Injection size: 0.5 L Carrier gas: Helium System suitability Sample: Standard solution [NOTEThe relative retention times for alcohol and npropyl alcohol are 1.0 and 1.5, respectively.] Suitability requirements Resolution: NLT 2 between alcohol and n-propyl alcohol Tailing factor: NMT 3.0 for the alcohol peak Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C2H5OH in the portion of Oral Rinse taken: Result = (RU/RS) (WS/WU) (100/L) = peak response ratio of alcohol to n-propyl alcohol from the Sample solution = peak response ratio of alcohol to n-propyl RS alcohol from the Standard solution = weight of dehydrated alcohol taken to prepare WS the Standard solution (g) = weight of Oral Rinse taken to prepare the Sample WU solution (g) L = labeled amount of alcohol Acceptance criteria: 90.0%115.0% RU IMPURITIES Organic Impurities PROCEDURE: LIMIT OF p-CHLOROANILINE Solution A, Solution B, Mobile phase, Diluent, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Standard solutions: Transfer 10 mg of p-chloroaniline to a 100-mL volumetric flask, add 2 mL of acetonitrile, swirl to dissolve, and dilute with Diluent to volume. Dilute this solution quantitatively, and stepwise if necessary, with Diluent to obtain Standard solutions having known concentrations of 1.5, 1.2, 0.6, and 0.3 g/mL of pchloroaniline. Sample solution: Transfer 10.0 mL of Oral Rinse to a 25mL volumetric flask, and dilute with Diluent to volume. Analysis Samples: Standard solutions and Sample solution Separately inject equal volumes (50 L) of the Standard solutions and Sample solution. Record the chromatograms, and measure the areas for the pchloroaniline peaks. Plot the peak responses from the Standard solutions versus the relevant concentrations, in g/mL. Draw the straight line best fitting the four plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of p-chloroaniline in the Sample solution. Calculate the quantity, in g/mL, of p-chloroaniline in the portion of Oral Rinse taken: Result = 2.5 C
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, at controlled room temperature. LABELING: The labeling indicates that the Oral Rinse is to be expectorated and not swallowed after rinsing. USP REFERENCE STANDARDS 11 USP Chlorhexidine Acetate RS USP Potassium Gluconate RS
Chlorhexidine Gluconate Solution

(Comment on this Monograph)id=m15620=Chlorhexidine Gluconate Solution=Chl-Cy-Monos.pdf) 897.76 C22H30Cl2N10 2C6H12O7 2,4,11,13-Tetraazatetradecanediimidamide, N,N-bis(4chlorophenyl)-3,12-diimino-, di-D-gluconate; 1,1-Hexamethylenebis[5-(p-chlorophenyl)biguanide] di-Dgluconate [18472-51-0]. DEFINITION Chlorhexidine Gluconate Solution is an aqueous solution of chlorhexidine gluconate. It contains NLT 19.0% and NMT 21.0% of C22H30Cl2N10 2C6H12O7 (w/v). IDENTIFICATION A. INFRARED ABSORPTION 197K Standard solution: 5 mg/mL of USP Chlorhexidine RS in 70% alcohol. Recrystallize this solution, and dry the crystals at 105 for 1 h. Sample solution: To 1 mL of Solution add 40 mL of water, and cool in ice. Add 10 N sodium hydroxide, dropwise with stirring, until the solution produces a red color on thiazole yellow paper, and add 1 mL in excess. Filter, wash the precipitate with water until the washings are free from alkali, recrystallize the residue from 70% alcohol, and dry the crystals at 105 for 1 h. B. The crystals obtained in Identification test A melt at 132136 (see Melting Range or Temperature 741). C. To 0.05 mL of Solution, add 5 mL of a solution of cetyltrimethylammonium bromide (1 in 100), 1 mL of 10 N sodium hydroxide, and 1 mL of bromine TS: a deep red color is produced. D. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 10 mg/mL of USP Potassium Gluconate RS Sample solution: Sample in water (1:5) Spray reagent: Dissolve 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid in a 100-mL volumetric flask. Add 1.0 g of ceric sulfate, swirl to dissolve, and dilute with 2 N sulfuric acid to volume. Application volume: 5 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3) Analysis Samples: Standard solution and Sample solution Develop the chromatogram in a solvent system until the solvent front has moved 10 cm from the point of spotting. Remove the plate from the chamber, and dry at 110 for 20 min. Allow to cool, and spray with Spray reagent. Heat the plate at 110 for 10 min.
USP 32
Acceptance criteria: The principal spot from the Sample solution corresponds in color, size, and RF value to that from the Standard solution. ASSAY PROCEDURE Diluent: 27.6 g of monobasic sodium phosphate in 1.5 L of water. Adjust with phosphoric acid to a pH of 3.0, and dilute with water to 2000 mL. Solution A: Dissolve 27.6 g of monobasic sodium phosphate and 10 mL of triethylamine in 1.5 L of water. Adjust with phosphoric acid to a pH of 3.0, and dilute with water to 2000 mL. Mix the resulting solution and acetonitrile (70:30). Solution B: Acetonitrile Mobile phase: Use variable mixtures of Solution A and Solution B as directed in the gradient table below.
Official Monographs / Chlorhexidine 19

= molecular weight of chlorhexidine acetate, 625.55 Acceptance criteria: 19.0%21.0% IMPURITIES Organic Impurities PROCEDURE 1 Solution A, Solution B, Mobile phase, and Diluent: Proceed as directed in the Assay. Sample stock solution: Transfer 5.0 mL of Solution to a 100-mL volumetric flask, and dilute with water to volume. Sample solution: 5.0 mL of the Sample stock solution to a 25-mL volumetric flask, and dilute with Diluent to volume. This solution contains 2 mg/mL of chlorhexidine gluconate. Reference solution A: 3.0 mL of the Sample solution to a 50-mL volumetric flask, and dilute with Diluent to volume. This solution contains 0.06 mg/mL of chlorhexidine gluconate. Reference solution B: 2.0 mL of Reference solution A to a 100-mL volumetric flask, and dilute with Diluent to volume. This solution contains 0.0012 mg/mL of chlorhexidine gluconate. System suitability solution: Transfer 10 mg of USP Chlorhexidine Related Compounds RS to a 10-mL volumetric flask. Dissolve in 2 mL of acetonitrile, and dilute with Diluent to volume. Chromatographic system: Proceed as directed in the Assay, except the Injection size and chromatograph are programmed as shown in the gradient table below.
Mr2
System suitability solution: 50 g/mL of USP Chlorhexidine Acetate RS and 1 g/mL of p-chloroaniline in Diluent Standard solution: 1 mg/mL of USP Chlorhexidine Acetate RS. Prepare 50 g/mL from the resulting solution with Diluent. Sample stock solution: Transfer 5.0 mL of Solution to a 250-mL volumetric flask, and dilute with water to volume. Sample solution: Transfer 5.0 mL of the Sample stock solution to a 250-mL volumetric flask, and dilute with Diluent. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 239 nm Column: 4.6-mm 25-cm; base-deactivated 5-m packing L1 Column temperature: 40 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 3.0 between chlorhexidine and pchloroaniline Relative standard deviation: NMT 2.0% from the chlorhexidine peak, and NMT 5.0% from the pchloroaniline peak Analysis Samples: Standard solution and Sample solution Calculate the percentage (w/v) of C22H30Cl2N10 2C6H12O7 in the portion of Solution taken: Result = (rU/rS) (0.25 CS) (Mr1/Mr2) 100 rU rS CS Mr1 = peak response area of chlorhexidine from the Sample solution = peak response area of chlorhexidine from the Standard solution = concentration of USP Chlorhexidine Acetate RS in the Standard solution (g/mL) = molecular weight of chlorhexidine gluconate, 897.76
Injection size: 20 L System suitability Sample: System suitability solution [NOTEThe relative retention times for the main related compound peak and chlorhexidine are 0.6 and 1.0, respectively.] Suitability requirements Resolution: The two peaks between the main related compound peak and the chlorhexidine peak should be at least partially resolved from each other and completely resolved from the chlorhexidine peak. Analysis Samples: Sample solution, Reference solution A, and Reference solution B Examine the chromatogram from the Sample solution. Acceptance criteria: The sum of the peak areas, other than chlorhexidine and any peak areas less than that obtained for chlorhexidine in the chromatogram from Reference solution B, is NMT the peak area for chlorhexidine in the chromatogram from Reference solution A (3.0%). PROCEDURE 2: LIMIT OF p-CHLOROANILINE Solution A, Solution B, Mobile phase, Diluent, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Standard solutions: Transfer 10 mg of p-chloroaniline to a 100-mL volumetric flask, add 2 mL of acetonitrile, swirl to dissolve, and dilute with Diluent to volume. Dilute a volume of this solution with Diluent to obtain Standard solutions having known concentrations of 1.5, 1.2, 0.6, and 0.3 g/mL of p-chloroaniline.
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Chlorhexidine / Official Monographs
USP 32
Sample stock solution: Transfer 100 mg of previously dried Chlorophyllin Copper Complex Sodium to a Kjeldahl flask. Add 2.0 mL of sulfuric acid, 1.0 mL of nitric acid, and 1.0 mL of hydrogen peroxide, and carefully heat under a fume hood until a light green color is obtained. [NOTEIf the solution has any hint of a brown tint, continue to add 0.5mL portions of nitric acid until a green color is obtained.] Cool, transfer the contents quantitatively to a 1000-mL volumetric flask with several portions of water, dilute the contents of the flask with water to volume, and mix. Sample solution: Transfer 10.0 mL of Sample stock solution to a 50-mL volumetric flask, and dilute with water to volume. Spectrometric conditions Mode: Atomic absorption spectrometer Lamp: Copper hollow-cathode lamp Flame: Airacetylene Analytical wavelength: Copper emission line of 324.8 nm Blank: Water Analysis: Determine the absorbances of the Standard solutions and the Sample solution. Plot the absorbances of the Standard solutions versus the concentration, in g/mL, of copper, and draw the straight line best fitting the four plotted points. From the graph so obtained, determine the concentration, C, in g/mL, of copper in the Sample solution. Calculate the percentage of copper in the portion of Chlorophyllin Copper Complex Sodium taken: Result = 500 (C/W) = weight of Chlorophyllin Copper Complex Sodium taken to prepare the Sample solution (in mg on the dried basis) LIMIT OF IONIC COPPER Copper stock solution and Standard solutions: Prepare as directed in the test for Content of total copper. Sample solution: Transfer 100 mg of Chlorophyllin Copper Complex Sodium to a 150-mL conical flask, add 75 mL of water, and shake by mechanical means for 3 min. Adjust with 1 N hydrochloric acid to a pH of 3.0, transfer the suspension thus obtained to a 100-mL volumetric flask, and dilute with water to volume. Filter this suspension, discarding the first 10 mL of the filtrate. Analysis: Proceed as directed for Analysis in the test for Content of total copper. Calculate the percentage of ionic copper in the portion of Chlorophyllin Copper Complex Sodium taken: Result = 10 (C/W) [NOTETerms are as defined in Content of total copper.] Acceptance criteria: NMT 0.25%, calculated on the dried basis CONTENT OF CHELATED COPPER: NLT 4.0% Analysis: Calculate the percentage of chelated copper in the portion of Chlorophyllin Copper Complex Sodium taken by subtracting the percentage of ionic copper found in the test for Limit of ionic copper from the percentage of total copper found in the test for Content of total copper, calculated on the dried basis. CONTENT OF SODIUM Standard stock solution: Dissolve 254.2 mg of sodium chloride, previously dried at 105 for 2 h, in 50 mL of water, transfer to a 1000-mL volumetric flask, and dilute with water to volume. Each mL of this solution contains 100 g of sodium. Standard solutions: Transfer to each of five 100-mL volumetric flasks 10 mL of a solution of a nonionic wetting agent (1 in 500). Dilute the contents of one of the flasks with water to volume to provide a blank. To the remaining flasks add, respectively, 2.5, 5.0, 10.0, and 15.0 mL of the W
Sample stock solution: Transfer 5.0 mL of Solution to a 100-mL volumetric flask, and dilute with water to volume. Sample solution: 10.0 mL of Standard stock solution to a 250-mL volumetric flask, and dilute with Diluent to volume. This solution contains 0.4 mg/mL of chlorhexidine gluconate. Analysis Samples: Separately inject equal volumes (50 L) of the Standard solutions and Sample solution. Record the chromatograms, and measure the areas for the pchloroaniline peaks. Plot the peak responses from the Standard solutions versus the relevant concentrations, in g/mL. Draw the straight line best fitting the four plotted points. From this graph, determine the concentration, C, in g/mL, of p-chloroaniline in the Sample solution. Calculate the quantity, in g/mL, of p-chloroaniline in the portion of Solution taken: Result = 500 C [NOTEIf the quantity so obtained is less than 150 g/mL, the tests may be repeated using a more appropriate dilution in the preparation of the Sample solution.] Acceptance criteria: NMT 500 g/mL SPECIFIC TESTS SPECIFIC GRAVITY 841: 1.061.07 PH 791: 5.57.0, when diluted 1 in 20 with water ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Chlorhexidine RS USP Chlorhexidine Acetate RS USP Chlorhexidine Related Compounds RS USP Potassium Gluconate RS
Chlorophyllin Copper Complex Sodium

(Comment on this Monograph)id=m15900=Chlorophyllin Copper Complex Sodium=Chl-Cy-Monos.pdf) DEFINITION Chlorophyllin Copper Complex Sodium contains sodium salts of copper-chelated chlorophyll derivatives. It contains no artificial coloring. IDENTIFICATION A. SPECTROPHOTOMETRY AND LIGHT-SCATTERING (in the visible region) 851 Sample solution: 10 g/mL Medium: pH 7.5 phosphate buffer, prepared by mixing 0.15 M dibasic sodium phosphate and 0.15 M monobasic potassium phosphate (21:4) Ratio: A405/A630, 3.03.9 OTHER COMPONENTS CONTENT OF TOTAL COPPER: NLT 4.25% Copper stock solution: Transfer 1.000 g of copper to a 1000-mL volumetric flask, dissolve in 20 mL of nitric acid, and dilute with 0.2 N nitric acid to volume. This solution contains 1000 g of copper per mL. [NOTEStore in a polyethylene bottle.] Standard solutions: Pipet 5.0 mL of Copper stock solution into a 500-mL volumetric flask, and dilute with water to volume. Transfer 5.0, 10.0, 15.0, and 20.0 mL, respectively, of this solution to separate 100-mL volumetric flasks, and dilute the contents of each flask with water to volume. These Standard solutions contain 0.5, 1.0, 1.5, and 2.0 g/mL of copper, respectively.
USP 32
Standard stock solution, dilute with water to volume, and mix. These Standard solutions contain 2.5, 5.0, 10.0, and 15.0 g/mL of sodium, respectively. Sample solution: Transfer 100 mg of Chlorophyllin Copper Complex Sodium to a 1000-mL volumetric flask, add 100 mL of a solution of nonionic wetting agent (1 in 500) and 400 mL of water, and shake by mechanical means for 5 min. Dilute with water to volume. Standard graph: Set the flame photometer for maximum transmission at a wavelength of 589 nm. Adjust the instrument to zero transmittance with the blank. Adjust the instrument to 100% transmittance with the most concentrated of the Standard solutions. Read the percentage of transmittance of the other Standard solutions, and plot the percentage of transmittance versus the concentration, in g/mL, of sodium. Analysis: Adjust the instrument as directed under Standard graph, read the percentage of transmittance of the Sample solution, and from the Standard graph read the concentration, C, in g/mL, of sodium in the Sample solution. Calculate the percentage of sodium in the portion of Chlorophyllin Copper Complex Sodium taken: Result = 100 (C/W) = weight of Chlorophyllin Copper Complex Sodium taken to prepare the Sample solution (mg) Acceptance criteria: 5%7%, calculated on the dried basis NITROGEN DETERMINATION, Method I 461: NLT 4.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 30%, calculated on the dried basis ARSENIC, Method II 211: NMT 3 ppm LEAD 251: NMT 10 ppm IRON 241: NMT 0.50% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Escherichia coli and Salmonella species. PH 791: 9.510.7, in a solution (1 in 100) LOSS ON DRYING 731: Dry at 105 for 2 h: it loses NMT 5% of its weight. TEST FOR FLUORESCENCE: Apply 10 L of a 1% solution of it on filter paper, allow to dry, and examine the area of application under long-wavelength UV light through a red optical filter: no fluorescence is visible. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. W
Official Monographs / Chloroprocaine 21

DEFINITION Chloroprocaine Hydrochloride contains NLT 98.0% and NMT 102.0% of C13H19ClN2O2 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 290 nm Medium: pH 4.5 buffer Prepare by dissolving 13.61 g of monobasic potassium phosphate in 750 mL of water, adjusting to a pH of 4.5 0.1 with potassium hydroxide solution (1 in 180), and diluting with water to 1000 mL. Sample solution: 10 g/mL Absorptivities: Do not differ by more than 3.0%, calculated on the dried basis. C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Mobile phase: 0.8 mg/mL of sodium 1-heptanesulfonate in water, acetonitrile, methanol, and glacial acetic acid (74:20:5:1) Solution A: 0.2 mg/mL of recrystallized 4-amino-2chlorobenzoic acid in methanol Standard solution: Transfer about 50 mg of USP Chloroprocaine Hydrochloride RS to a 50-mL volumetric flask containing 5.0 mL of Solution A, add 15 mL of methanol, and dilute with water to volume. This solution contains 1 mg of USP Chloroprocaine Hydrochloride RS and 0.02 mg of 4-amino-2-chlorobenzoic acid per mL. System suitability solution: Equal volumes of Solution A and Standard solution Sample solution: 100 mg of Chloroprocaine Hydrochloride into a 100mL volumetric flask. Dissolve in 40 mL of methanol, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for 4-amino-2chlorobenzoic acid and chloroprocaine are about 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5.0 between 4-amino-2-chlorobenzoic acid and chloroprocaine, System suitability solution. Relative standard deviation: NMT 1.0% from chloroprocaine, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C13H19ClN2O2 HCl in the portion of Chloroprocaine Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response of chloroprocaine in the Sample solution = peak response of chloroprocaine in the Standard solution = concentration of USP Chloroprocaine Hydrochloride RS in the Standard solution (mg/mL) = concentration of Chloroprocaine Hydrochloride in the Sample solution (mg/mL)
Chloroprocaine Hydrochloride
(Comment on this Monograph)id=m15920=Chloroprocaine Hydrochloride=Chl-Cy-Monos.pdf)
307.22 C13H19ClN2O2 HCl Benzoic acid, 4-amino-2-chloro-, 2-(diethylamino)ethyl ester, monohydrochloride; 2-(Diethylamino)ethyl 4-amino-2-chlorobenzoate monohydrochloride [3858-89-7].
CU
22
Chloroprocaine / Official Monographs

98.0%102.0%
USP 32
Analysis Samples: Standard solution and Sample solution Proceed as directed in the chapter. Allow the spots to dry, and develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution (RF 0.40). After viewing, spray the chromatogram with Spray reagent: violet-blue colored spots, characteristic of tertiary nitrogen compounds, are visible. ASSAY PROCEDURE Mobile phase: 0.8 mg/mL of sodium 1-heptanesulfonate in water, acetonitrile, methanol, and glacial acetic acid (74:20:5:1) Solution A: 0.2 mg/mL of recrystallized 4-amino-2chlorobenzoic acid in methanol Standard solution: 1 mg/mL of USP Chloroprocaine Hydrochloride RS and 0.02 mg/mL of 4-amino-2chlorobenzoic acid. Transfer 50 mg of USP Chloroprocaine Hydrochloride RS to a 50-mL volumetric flask containing 5.0 mL of Solution A, add 15 mL of methanol, and dilute with water to volume. System suitability solution: Equal volumes of Solution A and Standard solution Sample solution: Volume of injection, equivalent to 100 mg of chloroprocaine hydrochloride, to a 100-mL volumetric flask, add 40 mL of methanol, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 5 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for 4-amino-2chlorobenzoic acid and chloroprocaine are about 0.35 and 1.0, respectively.] Suitability requirements Resolution: NLT 5.0 between 4-amino-2-chlorobenzoic acid and chloroprocaine, System suitability solution Relative standard deviation: NMT 1.0% from chloroprocaine, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C13H19ClN2O2 HCl in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of chloroprocaine from the Sample solution = peak response of chloroprocaine from the rS Standard solution = concentration of USP Chloroprocaine CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of chloroprocaine CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% rU IMPURITIES Organic Impurities PROCEDURE Analysis: Using the chromatograms obtained as directed for the Assay, calculate the percentage of 4-amino-2-
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Analysis: Using the chromatograms obtained as directed in the Assay, calculate the percentage of 4-amino-2chlorobenzoic acid (C7H6ClNO2) in the portion of Chloroprocaine Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response of 4-amino-2-chlorobenzoic acid in the Sample solution = peak response of 4-amino-2-chlorobenzoic acid rS in the Standard solution = concentration of 4-amino-2-chlorobenzoic acid CS in the Standard solution (mg/mL) = concentration of Chloroprocaine Hydrochloride CU in the Sample solution (mg/mL) Acceptance criteria: NMT 0.625% of 4-amino-2chlorobenzoic acid SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 173176 ACIDITY: Dissolve 1.0 g in 25 mL of water, add 2 drops of methyl red TS, and titrate with 0.020 N sodium hydroxide: NMT 1.8 mL is required to produce a yellow color. LOSS ON DRYING 731: Dry 500 mg at 105 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chloroprocaine Hydrochloride RS rU
Chloroprocaine Hydrochloride Injection

(Comment on this Monograph)id=m15930=Chloroprocaine Hydrochloride Injection=Chl-Cy-Monos.pdf) DEFINITION Chloroprocaine Hydrochloride Injection is a sterile solution of Chloroprocaine Hydrochloride in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of chloroprocaine hydrochloride (C13H19ClN2O2 HCl). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: Dissolve 60 mg of USP Chloroprocaine Hydrochloride RS in 10 mL of water in a 60-mL separator. Add 5 mL of dilute ammonium hydroxide (4 in 10), and immediately extract with four 10-mL portions of chloroform, passing the extracts through cotton filters into a 50-mL volumetric flask. Dilute with chloroform to volume. Sample solution: In a 60-mL separator, mix a volume of Injection, equivalent to 60 mg of chloroprocaine hydrochloride, with sufficient water to obtain 10 mL of solution. Add 5 mL of dilute ammonium hydroxide (4 in 10), and immediately extract with four 10-mL portions of chloroform, passing the extracts through cotton filters into a 50-mL volumetric flask. Dilute with chloroform to volume. Spray reagent: Iodoplatinate TS Application volume: 10 L Developing solvent system: Chloroform and methanol (4:1) Chromatographic chamber: Allow the system to equilibrate for 15 min with the Developing solvent system.
USP 32
chlorobenzoic acid (C7H6ClNO2) in the chloroprocaine hydrochloride contained in the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of 4-amino-2-chlorobenzoic acid from the Sample solution = peak response of 4-amino-2-chlorobenzoic acid rS from the Standard solution = concentration of 4-amino-2-chlorobenzoic acid CS in the Standard solution (mg/mL) CU = nominal concentration of 4-amino-2chlorobenzoic acid in the Sample solution (mg/mL) Acceptance criteria: NMT 3.0% SPECIFIC TESTS PH 791: 2.74.0 INJECTIONS 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Chloroprocaine Hydrochloride RS rU
Official Monographs / Chloroquine 23

B. ULTRAVIOLET ABSORPTION 197U Medium: Dilute hydrochloric acid (1 in 1000) Solution: 10 g/mL Acceptance criteria: The ratio A343/A329 is 1.001.15. ASSAY Change to read: PROCEDURE Sample solution: Dissolve 250 mg of Chloroquine in 50 mL of vglacial acetic acidvUSP32. Add crystal violet TS. Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 15.99 mg of C18H26ClN3. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
NMT 0.2%
Chloroquine
(Comment on this Monograph)id=m15980=Chloroquine=ChlCy-Monos.pdf)
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 8792 LOSS ON DRYING 731: Dry a sample at 105 for 2 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Chloroquine Phosphate RS
Chloroquine Hydrochloride Injection

319.87 C18H26ClN3 1,4-Pentanediamine, N4-(7-chloro-4-quinolinyl)-N1,N1-diethyl-; 7-Chloro-4-[[4-(diethylamino)-1-methylbutyl]amino]quinoline [54-05-7]. DEFINITION Chloroquine contains NLT 98.0% and NMT 102.0% of C18H26ClN3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION Standard solution: Dissolve 50 mg of USP Chloroquine Phosphate RS in 25 mL of 0.1 N hydrochloric acid. Add 2 mL 1 N sodium hydroxide and 4 mL of chloroform, and shake for 2 min. Centrifuge if necessary to clarify the lower phase, and pass through a dry filter. Sample solution: 8.75 mg/mL in chloroform, passed through a dry filter Analysis: Determine the absorption of the Standard solution and Sample solution without delay in 1-mm cells between 7 and 15 m, using chloroform in a matched cell as a blank. Acceptance criteria: The IR absorption spectrum of the Sample solution so obtained exhibits maxima only at the same wavelengths as that of the Standard solution. (Comment on this Monograph)id=m16030=Chloroquine Hydrochloride Injection=Chl-Cy-Monos.pdf) C18H26ClN3 2HCl 392.79 1,4-Pentanediamine, N4-(7-chloro-4-quinolinyl)-N1,N1-diethyl-, dihydrochloride; 7-(Chloro-4-4-diethylamino)-1-methylbutylaminoquinoline dihydrochloride [3545-67-3]. DEFINITION Chloroquine Hydrochloride Injection is a sterile solution of Chloroquine in Water for Injection prepared with the aid of Hydrochloric Acid. It contains NLT 47.5 mg and NMT 52.5 mg of C18H26ClN3 2HCl in each mL. IDENTIFICATION A. The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of the Standard solution concomitantly measured, as employed for measurement of absorbance in the Assay. The ratio A343/A329 is between 1.00 and 1.15. B. PROCEDURE Sample solution: 20 mL of a 1 mg/mL solution of chloroquine hydrochloride from Injection diluted with water
24
Chloroquine / Official Monographs
USP 32
IDENTIFICATION A. It meets the requirements under IdentificationOrganic Nitrogenous Bases 181, chloroform being substituted for carbon disulfide in the test. B. ULTRAVIOLET ABSORPTION 197U Medium: Dilute hydrochloric acid (1 in 1000) Solution: 10 g/mL Ratio: A343/A329, 1.001.15 ASSAY PROCEDURE Standard solution: 20 mg/mL of USP Chloroquine Phosphate RS in water. Prepare 10 g/mL from the resulting solution with dilute hydrochloric acid (1 in 1000). Sample solution: 20 mg/mL of Chloroquine Phosphate in water. Prepare 10 g/mL from the resulting solution with dilute hydrochloric acid (1 in 1000). Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 343 nm Cell: 1 cm Blank: Dilute hydrochloric acid (1 in 1000) Analysis Samples: Standard solution and Sample solution Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C18H26ClN3 2H3PO4 in the portion of Chloroquine Phosphate taken: Result = (AU/AS) (CS/CU) 100 AU = absorbance of the Sample solution AS = absorbance of the Standard solution = concentration of the Standard solution (g/mL) CS CU = concentration of the Sample solution (g/mL) Acceptance criteria: 98.0%102.0% SPECIFIC TESTS LOSS ON DRYING 731: 2.0% of its weight. Dry at 105 for 16 h: it loses NMT
Analysis: Add 5 mL of trinitrophenol TS. Acceptance criteria: A yellow precipitate is formed. Filter and wash the precipitate with water until the last washing is colorless, and dry over silica gel: the precipitate melts between 205 and 210. [CautionPicrates may explode.] C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Diluent 1: Dilute hydrochloric acid (1 in 1000) Diluent 2: Dilute hydrochloric acid (1 in 100) Standard solution: 10 g/mL of USP Chloroquine Phosphate RS in Diluent 1 Sample stock solution: From a volume of Injection, equivalent to 150 mg of chloroquine hydrochloride, to a 1L volumetric flask and dilute with water to volume Sample solution: 5.0 mL of Sample stock solution, and 10.0 mL of Diluent 2, and then dilute with water to 100.0 mL Spectrometric conditions Mode: UV spectrophotometry Analytical wavelength: 343 nm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the concentration, in mg/mL, of C18H26ClN3 2HCl in the portion of Injection taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2)/L = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Chloroquine Phosphate RS in the Standard solution (g/mL) = nominal concentration of chloroquine CU hydrochloride in the Sample solution (g/mL) = molecular weight of C18H26ClN3 2HCl, 392.79 Mr1 = molecular weight of C18H26ClN3 2H3PO4, 515.86 Mr2 L = label claim (mg/mL) Acceptance criteria: 47.552.5 mg/mL SPECIFIC TESTS PH 791: 5.56.5 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.7 USP Endotoxin Unit/mg chloroquine hydrochloride. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Chloroquine Phosphate RS USP Endotoxin RS AU AS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chloroquine Phosphate RS
Chloroquine Phosphate Tablets

(Comment on this Monograph)id=m16110=Chloroquine Phosphate Tablets=Chl-Cy-Monos.pdf) DEFINITION Chloroquine Phosphate Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of C18H26ClN3 2H3PO4. IDENTIFICATION A. The UV absorption spectrum of the solution employed for measurement of absorbance in the Assay exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Chloroquine Phosphate RS, concomitantly measured. The ratio A343/A329 is between 1.00 and 1.15. B. To 20-mL filtered solution of 1 mg/mL of Chloroquine Phosphate from finely powdered Tablets in water, add 5 mL of trinitrophenol TS: a yellow precipitate is formed. Filter, wash the precipitate with water until the last washing is
Chloroquine Phosphate
(Comment on this Monograph)id=m16080=Chloroquine Phosphate=Chl-Cy-Monos.pdf) 515.86 C18H26ClN3 2H3PO4 1,4-Pentanediamine, N4-(7-chloro-4-quinolinyl)-N1,N1-diethyl-, phosphate (1:2); 7-Chloro-4-[[4-(diethylamino)-1-methylbutyl]amino]quinoline phosphate (1:2) [50-63-5]. DEFINITION Chloroquine Phosphate contains NLT 98.0% and NMT 102.0% of C18H26CIN3 2H3PO4, calculated on the dried basis.
USP 32
colorless, and dry over silica gel: the precipitate melts between 205 and 210. [CAUTIONPicrates may explode.] ASSAY PROCEDURE Standard solution: 10 g/mL of USP Chloroquine Phosphate RS in dilute hydrochloric acid (1 in 1000) Sample solution: Transfer the equivalent of 800 mg of chloroquine phosphate from powdered Tablets (NLT 20), to a 200-mL volumetric flask, add 100 mL of water, and shake by mechanical means for 20 min. Add water to volume, and filter, discarding the first 50 mL of the filtrate. Pipet 50 mL of the clear filtrate into a 250-mL separator, add 5 mL of 6 N ammonium hydroxide, agitate, and extract the liberated chloroquine with five 25-mL portions of chloroform. Wash the combined chloroform extracts with 10 mL of water, and extract the water washing with 10 mL of chloroform. Evaporate the combined chloroform extracts on a steam bath to 10 mL, then add 50 mL of dilute hydrochloric acid (1 in 250), and continue heating on the steam bath until the odor of chloroform is no longer perceptible. Transfer the solution to a 200-mL volumetric flask, wash the evaporating vessel with portions of dilute hydrochloric acid (1 in 1000), adding the washings to the volumetric flask, and add more of the same dilute hydrochloric acid to volume. Dilute this solution quantitatively and stepwise to obtain a concentration of about 10 g/mL with dilute hydrochloric acid (1 in 1000). Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 343 nm Cell: 1 cm Blank: Dilute hydrochloric acid (1 in 1000) Analysis Samples: Standard solution and Sample solution Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C18H26ClN3 2H3PO4 in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of chloroquine phosphate in the Sample solution (g/mL) Acceptance criteria: 93.0%107.0% AU AS CS CU PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 100 rpm Time: 45 min Detector: UV 343 nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Chloroquine Phosphate RS in Medium Tolerances: NLT 75% (Q) of the labeled amount of C18H26ClN3 2H3PO4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chloroquine Phosphate RS = = = =
Official Monographs / Chlorothiazide 25
Chlorothiazide
(Comment on this Monograph)id=m16210=Chlorothiazide=ChlCy-Monos.pdf)
C7H6ClN3O4S2 295.73 2 H-1,2,4-Benzothiadiazine-7-sulfonamide, 6-chloro-, 1,1dioxide; 6-Chloro-2 H-1,2,4-benzothiadiazine-7-sulfonamide 1,1-dioxide [58-94-6]. DEFINITION Chlorothiazide contains NLT 98.0% and NMT 102.0% of C7H6ClN3O4S2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M: Previously dried at 105 for 1 h B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 292 nm Medium: Sodium hydroxide solution (1 in 250) Sample solution: 10 g/mL Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0% ASSAY PROCEDURE [NOTEThe Standard solution and Sample solution should be injected immediately upon preparation.] Mobile phase: Acetonitrile and 0.1 M monobasic sodium phosphate (1:9), adjust with phosphoric acid to a pH of 3.0 0.1 System suitability solution: 0.15 mg/mL of USP Chlorothiazide RS and 1.5 g/mL of USP Benzothiadiazine Related Compound A RS in Mobile phase Standard solution: 0.15 mg/mL of USP Chlorothiazide RS in Mobile phase [NOTEA volume of acetonitrile not exceeding 10% of the total volume of solution may be used to dissolve the reference standard.] Sample solution: 0.15 mg/mL of Chlorothiazide in Mobile phase [NOTEA volume of acetonitrile not exceeding 10% of the total volume of solution may be used to dissolve the sample.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for benzothiadiazine related compound A and chlorothiazide are about 0.9 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.5 between benzothiadiazine related compound A and chlorothiazide, System suitability solution Relative standard deviation: NMT 1.5%, Standard solution
26
Chlorothiazide / Official Monographs

USP Chlorothiazide RS
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6ClN3O4S2 in the portion of Chlorothiazide taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlorothiazide RS in the Standard solution (mg/mL) CU = nominal concentration of Chlorothiazide in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE: Dissolve 1.00 g in a mixture of 10 mL of water and 10 mL of sodium hydroxide solution (1 in 10). Cool in an ice bath, and add 20 mL of water and 5 mL of nitric acid. A flocculent, white precipitate is formed. Titrate potentiometrically with 0.050 N silver nitrate, using a silversilver chloride electrode system. Acceptance criteria: NMT 0.28 mL (0.05%) SELENIUM 291: NMT 30 ppm HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Mobile phase, System suitability solution, Sample solution and Chromatographic system: Proceed as directed in the Assay. Standard solution: 1.5 g/mL of USP Benzothiadiazine Related Compound A RS in Mobile phase Analysis Samples: Standard solution and Sample solution Calculate the percentage of benzothiadiazine related compound A in the portion of Chlorothiazide taken: Result = (rU/rS) (CS/CU) 100 = peak response of benzothiadiazine related compound A from the Sample solution rS = peak response of benzothiadiazine related compound A from the Standard solution CS = concentration of USP Benzothiadiazine Related Compound A RS in the Standard solution (g/mL) = nominal concentration of benzothiadiazine CU related compound A in the Sample solution (g/mL) Acceptance criteria: NMT 1.0% rU SPECIFIC TESTS LOSS ON DRYING 731: 1.0% of its weight. Dry at 105 for 1 h: it loses NMT
Chlorothiazide Oral Suspension

(Comment on this Monograph)id=m16240=Chlorothiazide Oral Suspension=Chl-Cy-Monos.pdf) DEFINITION Chlorothiazide Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of chlorothiazide (C7H6ClN3O4S2). IDENTIFICATION The UV absorption spectrum of the solution of chlorothiazide prepared from Oral Suspension as directed in the Assay exhibits maxima and minima at the same wavelengths as that of a solution of USP Chlorothiazide RS, prepared as directed in the Assay, concomitantly measured. ASSAY PROCEDURE Standard solution: 10 g/mL of USP Chlorothiazide RS in sodium hydroxide solution (1 in 250) Sample stock solution: Equivalent to 1 mg/mL of chlorothiazide from Oral Suspension, in sodium hydroxide solution (1 in 250) Sample solution: 10.0 mL of the resulting solution diluted with hydrochloric acid (1 in 100) to 100.0 mL. Transfer 50.0 mL of the resulting solution to a 125-mL separator, and wash with two 25-mL portions of chloroform, discarding the washings. Transfer 10.0 mL of the washed solution to a 100mL volumetric flask, and dilute with sodium hydroxide solution (1 in 250) to volume. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 292 nm Cell: 1 cm Blank: Sodium hydroxide solution (1 in 250) Analysis Samples: Standard solution and Sample solution Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C7H6ClN3O4S2 in each mL of the Oral Suspension taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of chlorothiazide in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% AU AS CS CU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for Oral Suspension packaged in single-unit containers DELIVERABLE VOLUME 698: Meets the requirements for Oral Suspension packaged in multiple-unit containers = = = =
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at 25; excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Benzothiadiazine Related Compound A RS
USP 32
SPECIFIC TESTS PH 791: 3.24.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Chlorothiazide RS CS
Official Monographs / Chlorothiazide 27

= concentration of USP Chlorothiazide RS in the Standard solution (mg/mL) = nominal concentration of chlorothiazide in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.05 M pH 8.0 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL Apparatus 2: 75 rpm Time: 60 min Detector: UV 294 nm Standard solution: USP Chlorothiazide RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Tolerances: NLT 75% (Q) of the labeled amount of C7H6ClN3O4S2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chlorothiazide RS
Chlorothiazide Tablets
(Comment on this Monograph)id=m16250=Chlorothiazide Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlorothiazide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C7H6ClN3O4S2. IDENTIFICATION A. The chromatogram of the Sample solution, obtained as directed in the Assay, exhibits a peak for chlorothiazide, the retention time of which corresponds to that exhibited by the Standard solution. B. IDENTIFICATION TESTSGENERAL, Sulfite 191: Powder 1 Tablet, and fuse it with a pellet of sodium hydroxide: the ammonia fumes produced turn moistened red litmus paper blue, and the residue meets the requirements. ASSAY PROCEDURE Mobile phase: Prepare a solution of 0.08 M monobasic sodium phosphate (adjusted with phosphoric acid to a pH of 2.9 0.1) and methanol (95:5). Standard solution: 0.5 mg/mL. Transfer 25 mg of USP Chlorothiazide RS to a 50-mL volumetric flask, add 5 mL of 0.05 M monobasic sodium phosphate solution, followed by 10 mL of acetonitrile to the flask, and sonicate with occasional shaking for 3 min. Dilute with water to volume and filter. Sample solution: Equivalent to 250 mg of chlorothiazide from finely powdered Tablets (NLT 20) to a 500-mL volumetric flask. Add 50 mL of 0.05 M monobasic sodium phosphate solution, and shake by mechanical means for 15 min, followed by sonication for 2 min. Add 100 mL of acetonitrile, sonicate for 3 min, and dilute with water to volume. [NOTEPrepare fresh daily.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1, and fitted with a guard column Flow rate: 2 mL/min Injection size: 15 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 4.3 Tailing factor: NMT 2.0 for chlorothiazide Column efficiency: NLT 1300 theoretical plates for chlorothiazide Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H6ClN3O4S2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
Chlorothiazide Sodium for Injection

(Comment on this Monograph)id=m16280=Chlorothiazide Sodium for Injection=Chl-Cy-Monos.pdf) C7H5ClN3NaO4S2 317.71 2H-1,2,4-Benzothiadiazine-7-sulfonamide, 6-chloro-, 1,1-dioxide, monosodium salt; 6-Chloro-2H-1,2,4-benzothiadiazine-7-sulfonamide 1,1-dioxide monosodium salt [7085-44-1]. DEFINITION Chlorothiazide Sodium for Injection is a sterile, freeze-dried mixture of Chlorothiazide Sodium (prepared by the neutralization of Chlorothiazide with the aid of Sodium Hydroxide) and Mannitol. It contains chlorothiazide sodium (C7H5ClN3NaO4S2) equivalent to NLT 93.0% and NMT 107.0% of the labeled amount of chlorothiazide (C7H6ClN3O4S2). IDENTIFICATION ULTRAVIOLET ABSORPTION 197U Medium: Sodium hydroxide solution (1 in 250) Sample solution: 10 g/mL ASSAY PROCEDURE Standard solution: 10 g/mL of USP Chlorothiazide RS in sodium hydroxide solution (1 in 250) Sample solution: Equivalent to 10 g/mL of chlorothiazide from Injection, in sodium hydroxide solution (1 in 250) Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 292 nm Cell: 1 cm Blank: Sodium hydroxide solution (1 in 250) Analysis Samples: Standard solution and Sample solution Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C7H6ClN3O4S2 in the portion of Chlorothiazide Sodium for Injection taken: Result = (AU/AS) (CS/CU) 100
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Chlorothiazide / Official Monographs

AU = absorbance of the Sample solution = absorbance of the Standard solution AS = concentration of the Standard solution (g/mL) CS = concentration of the Sample solution (g/mL) CU Acceptance criteria: 93.0%107.0%
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Temperature Column: 210 Injector: 210 Detector: 210 Carrier gas: Dry nitrogen Flow rate: 30 mL/min Injection size: 2 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between the p-chlorophenol peak and the chloroxylenol peak Tailing factor: NMT 1.5 for the chloroxylenol peak Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9ClO in the portion of Chloroxylenol taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the chloroxylenol peak to that of the p-chlorophenol peak from the Sample solution = peak response ratio of the chloroxylenol peak to RS that of the p-chlorophenol peak from the Standard solution CS = concentration of USP Chloroxylenol RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: NLT 98.5% of C8H9ClO RU IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% IRON 241 Sample: 0.10 g Analysis: Transfer the Sample to a suitable crucible, add 5 drops of sulfuric acid, and ignite at a low heat until thoroughly ashed. Add to the carbonized mass 10 drops of sulfuric acid, and heat cautiously until white fumes are no longer evolved. Ignite, preferably in a muffle furnace, at 500600, until the carbon is completely burned off. Cool, add 4 mL of 6 N hydrochloric acid, cover, digest on a steam bath for 15 min, uncover, and slowly evaporate on a steam bath to dryness. Moisten the residue with 1 drop of hydrochloric acid, add 10 mL of hot water, and digest for 2 min. Dilute with water to 25 mL. Filter, if necessary, rinse the crucible and the filter with 10 mL of water, combining the filtrate and rinsing in a 50-mL color-comparison tube, add 2 mL of hydrochloric acid, dilute with water to 47 mL, and mix. Acceptance criteria: NMT 0.01% Organic Impurities PROCEDURE Standard solution: 0.08 mg/mL each of 3,5dimethylphenol and USP Chloroxylenol Related Compound A RS in chloroform Sample solution: 40.0 mg/mL of chloroxylenol in chloroform Chromatographic system (See Chromatography 621, System Suitability.)
PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905:
SPECIFIC TESTS PH 791: 9.210.0, in a solution prepared as directed in the labeling BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.3 USP Endotoxin Unit/mg of chlorothiazide sodium. INJECTIONS, Constituted Solutions 1: At the time of use, it meets the requirements. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Chlorothiazide RS USP Endotoxin RS
Chloroxylenol
(Comment on this Monograph)id=m16420=Chloroxylenol=ChlCy-Monos.pdf)
C8H9ClO Phenol, 4-chloro-3,5-dimethyl-; 4-Chloro-3,5-xylenol [88-04-0]. DEFINITION Chloroxylenol contains NLT 98.5% of C8H9ClO. IDENTIFICATION INFRARED ABSORPTION 197K
156.61
ASSAY PROCEDURE Internal standard solution: 0.8 mg/mL of p-chlorophenol in chloroform Standard solution: 1 mg/mL of USP Chloroxylenol RS in the Internal standard solution Sample solution: 1 mg/mL of Chloroxylenol in the Internal standard solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 1.8-m column packed with 3% phase G16 on support S1A
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Mode: GC Detector: Flame ionization Column: 4-mm 1.8-m column packed with 3% phase G16 on support S1A Temperature Column: 180 Injector: 200 Detector: 200 Carrier gas: Dry nitrogen Flow rate: 30 mL/min Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 4.5 between 3,5-dimethylphenol and chloroxylenol related compound A Relative standard deviation: NMT 10% Analysis Samples: Standard solution and Sample solution Calculate the percentages of 3,5-dimethylphenol (C8H10O) and chloroxylenol related compound A (C8H9ClO) in the portion of Chloroxylenol taken: Result = (rU/rS) 100 = peak area of the appropriate analyte from the Sample solution = peak area of the appropriate analyte from the rS Standard solution Calculate the percentage of each impurity in the portion of Chloroxylenol taken: rU Result = (rU/rT) 100 rU = area of each peak from the Sample solution, excluding that of the main chloroxylenol peak, the 3,5-dimethylphenol peak, and the chloroxylenol related compound A peak rT = sum of the areas of all the peaks Calculate the percentage of total impurities in the portion of Chloroxylenol taken: Result = (rT/rS) 100 = sum of the areas of all the peaks from the Sample solution, excluding that of the main chloroxylenol peak = sum of the areas of all the peaks from the rS Sample solution Acceptance criteria 3,5-Dimethylphenol: NMT 0.2% Chloroxylenol related compound A: NMT 0.2% Any individual impurity: NMT 0.5% Total impurities: NMT 1.5% rT SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 114116 WATER DETERMINATION, Method I 921: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chloroxylenol RS
Official Monographs / Chlorpheniramine 29

USP Chloroxylenol Related Compound A RS
Chlorpheniramine Maleate
(Comment on this Monograph)id=m16450=Chlorpheniramine Maleate=Chl-Cy-Monos.pdf)
390.86 C16H19ClN2 C4H4O4 2-Pyridinepropanamine, -(4-chlorophenyl)-N,N-dimethyl-, (Z)-2-butenedioate (1:1); 2-[p-Chloro--[2-(dimethylamino)ethyl]benzyl] pyridine maleate (1:1) [113-92-8]. DEFINITION Chlorpheniramine Maleate contains NLT 98.0% and NMT 100.5% of C16H19ClN2 C4H4O4, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample solution: 500 mg in 20 mL of glacial acetic acid and 2 drops of crystal violet TS Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 19.54 mg of C16H19ClN2 C4H4O4. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Sample solution: 40 mg/mL of Chlorpheniramine Maleate in methylene chloride Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization detector Column: 1.2-m 4-mm glass column containing 3% phase G3 on support S1AB Temperature Column: 190 Injector: 250 Detector: 250 Carrier gas: Dry helium Flow rate: Adjust to obtain a retention time of 45 min for the main peak.
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Chlorpheniramine / Official Monographs

Injection size: 1 L System suitability Sample: Sample solution Suitability requirements Tailing factor: NMT 1.8 Analysis Sample: Sample solution Record the chromatogram for a total time of NLT twice the retention time of the chlorpheniramine peak, and measure the areas of the peaks. Acceptance criteria: The total relative area of all extraneous peaks (except that of the solvent peak and maleic acid, if observed) does not exceed 2.0%.
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acid (1 in 100) to volume, and filter. Transfer 10.0 mL of the filtrate to a 50-mL volumetric flask, and dilute with dilute hydrochloric acid (1 in 100) to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 261 nm Column: 3.9-mm 15-cm; 10-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 900 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of the label claim of C16H19ClN2 C4H4O4 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of the Capsules in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 1. Medium: Water; 500 mL Apparatus 1: 100 rpm Times: 1.5, 6.0, and 10.0 h Analysis: Determine the amount of C16H19ClN2 C4H4O4 dissolved by using the procedure set forth in the Assay, using a filtered portion of the solution under test in comparison with a Standard solution having a known concentration of USP Chlorpheniramine Maleate RS in the same Medium. Tolerances: The percentages of the labeled amount of C16H19ClN2 C4H4O4 dissolved at the times specified conform to the acceptance table below.
Time (h) 1.5 6.0 10.0 Amount Dissolved 15%40% 50%80% NLT 70%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 130135 LOSS ON DRYING 731: Dry at 105 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS
Chlorpheniramine Maleate ExtendedRelease Capsules

(Comment on this Monograph)id=m16460=Chlorpheniramine Maleate Extended-Release Capsules=Chl-Cy-Monos.pdf) DEFINITION Chlorpheniramine Maleate Extended-Release Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of chlorpheniramine maleate (C16H19ClN2 C4H4O4). IDENTIFICATION A. The retention time of the chlorpheniramine peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 1.2 mg/mL USP Chlorpheniramine Maleate RS in methanol and water (1:1) Sample solution: Transfer the contents of 1 Capsule to a 10-mL volumetric flask, add 5 mL of methanol, and insert the stopper into the flask. Sonicate this solution for 10 min, dilute with water to volume, mix, and filter. Application volume: 10 L Developing solvent system: Ethyl acetate, methanol, and ammonium hydroxide (20:1:1) Analysis: Proceed as directed for Chromatography 621, Thin-Layer Chromatography, and examine the plate under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: 2.0 g of sodium perchlorate and 2.0 mL of triethylamine/L of methanol and water (650:350) Standard solution: 0.12 mg/mL of USP Chlorpheniramine Maleate RS in dilute hydrochloric acid (1 in 100) Sample solution: Weigh and mix the contents of NLT 20 Capsules. Transfer a portion of the mixture, equivalent to 120 mg of chlorpheniramine maleate, to a 200-mL volumetric flask. Add 100 mL of dilute hydrochloric acid (1 in 100), bring to a boil on a hot plate, and continue boiling moderately for 5 min. Cool, dilute with dilute hydrochloric
rU rS CS CU
= = = =
Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Proceed as directed for Apparatus 1 and Apparatus 2, Delayed-Release Dosage Forms, Method B, Procedure. Medium: Prepare as directed under Method B, except to use 900 mL of media. Operate the apparatus for 1 h in the Acid Stage, and use the acceptance criteria given in Tolerances. Operate the apparatus for 6 h in the Buffer Stage, except to use 900 mL of simulated intestinal fluid TS without enzyme, and use the acceptance criteria given in Tolerances. Apparatus 2: 50 rpm Times: 1.0, 3.0, and 7.0 h Analysis: Proceed as directed in Test 1. Tolerances: The percentages of the labeled amount of C16H19ClN2 C4H4O4 dissolved at the times specified conform to the acceptance table below.
USP 32
Time (h) 1.0 3.0 7.0

USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS USP Endotoxin RS
Amount Dissolved 30%60% 55%85% NLT 70%
Chlorpheniramine Maleate Oral Solution

(Comment on this Monograph)id=m16480=Chlorpheniramine Maleate Oral Solution=Chl-Cy-Monos.pdf) DEFINITION Chlorpheniramine Maleate Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of chlorpheniramine maleate (C16H19ClN2 C4H4O4). IDENTIFICATION A. PROCEDURE Sample solution: Evaporate the remaining extract from the Assay on a steam bath to a small volume, then transfer it to a smaller, more suitable vessel, and evaporate it just to the point where hexane vapors are no longer perceptible. Transfer the oily residue, with the aid of four 3-mL portions of dimethylformamide, to a suitable glass-stoppered graduated cylinder, and dilute with dimethylformamide to 15.0 mL. Acceptance criteria: The optical rotation of the solution so obtained, in a 100-mm tube, after correcting for the blank, is NMT +0.01 (distinction from dexchlorpheniramine maleate). B. ULTRAVIOLET ABSORPTION 197U Sample solution: The solution used for measurement of absorbance in the Assay ASSAY PROCEDURE Standard solution: 0.4 mg/mL of USP Chlorpheniramine Maleate RS Sample solution: Oral Solution Analysis: Transfer 10 mL each of the Standard solution and the Sample solution into two different separators. Treat each solution as follows. Add 10 mL of sodium hydroxide solution (1 in 10), and extract with two 50-mL portions of solvent hexane. Combine the extracts in a second separator, wash with 10 mL of sodium hydroxide solution (1 in 250), and discard the washing. Extract the hexane solution with two 40-mL portions of dilute hydrochloric acid (1 in 100), collect the extracts in a 100-mL volumetric flask, and add the same dilute acid to volume. Wash 50-mL portions of each solution, and of dilute hydrochloric acid (1 in 100), respectively, with three 30-mL portions of chloroform and then with 50 mL of solvent hexane, and discard the washings. Filter the acid phases through paper, discarding the first few mL of each filtrate. Spectrometric conditions Analytical wavelength: 264 nm Cell: 1 cm Blank: Extracted acid Calculate the percentage of C16H19ClN2 C4H4O4 taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = = = = absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of chlorpheniramine maleate in the Sample solution (mg/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label the Capsules to indicate the Dissolution Test with which the product complies. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS
Chlorpheniramine Maleate Injection

(Comment on this Monograph)id=m16470=Chlorpheniramine Maleate Injection=Chl-Cy-Monos.pdf) DEFINITION Chlorpheniramine Maleate Injection is a sterile solution of Chlorpheniramine Maleate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C16H19ClN2 C4H4O4. IDENTIFICATION A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181: Dilute a volume of the Injection, equivalent to 50 mg of chlorpheniramine maleate, with dilute hydrochloric acid (1 in 1000) to 25 mL, and proceed as directed under the chapter beginning with Transfer the liquid to a separator. The Injection meets the requirements of the test. B. Evaporate a volume of the Injection, equivalent to 25 mg of chlorpheniramine maleate, on a steam bath to dryness, and dry the residue at 105 for 1 h: it melts between 128 and 135. ASSAY PROCEDURE Standard solution: 1.25 mg/mL of USP Chlorpheniramine Maleate RS in dilute sulfuric acid (1 in 350). Treat 20 mL of this solution the same as the portion of Injection being assayed. Sample solution: Proceed with Injection as directed under Salts of Organic Nitrogenous Bases 501. Analysis Mode: UV 264 nm Calculate the percentage of C16H19ClN2 C4H4O4 in the portion of Injection taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of chlorpheniramine maleate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: It contains NMT 8.8 USP Endotoxin Units/mg of chlorpheniramine maleate. PH 791: 4.05.2 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, protected from light. AU AS CS CU = = = =
32

90.0%110.0% If present, 6.0%8.0% Acceptance criteria: 90.0%110.0%
USP 32
OTHER COMPONENTS ALCOHOL DETERMINATION 611:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS
Chlorpheniramine Maleate Tablets

(Comment on this Monograph)id=m16490=Chlorpheniramine Maleate Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlorpheniramine Maleate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C16H19ClN2 C4H4O4. IDENTIFICATION INFRARED ABSORPTION 197M Sample solution: 1.25 mg/mL of chlorpheniramine maleate from a portion of powdered Tablets in dilute hydrochloric acid (1 in 100) Standard solution: 1.25 mg/mL of USP Chlorpheniramine Maleate RS in dilute hydrochloric acid (1 in 100) Analysis: Treat each solution as follows. Render alkaline, to a pH of about 11, with sodium hydroxide solution (1 in 10). Extract with two 50-mL portions of solvent hexane, collect the extracts in a beaker, and evaporate to dryness. Prepare a mineral oil dispersion of the residue so obtained and determine the infrared absorption spectrum of the preparation in the region between 2 and 12 m. Acceptance criteria: The spectrum of the Sample solution exhibits maxima only at the same wavelengths as that of the Standard solution. ASSAY PROCEDURE Standard solution: 0.2 mg/mL of USP Chlorpheniramine Maleate RS in dilute hydrochloric acid (1 in 100), and treat 20.0 mL of this solution the same as the Sample solution in dilute hydrochloric acid (1 in 100) of the portion of Tablets taken. Sample solution: Using a portion of powdered Tablets equivalent to 4 mg of chlorpheniramine maleate, proceed as directed under Salts of Organic Nitrogenous Bases 501, but using dilute hydrochloric acid (1 in 100) instead of the dilute sulfuric acid (1 in 350), and dilute sulfuric acid (1 in 70), and using solvent hexane instead of the ether, and diluting 10 mL of the Sample solution with dilute hydrochloric acid (1 in 100) to 25.0 mL. Analysis Mode: UV 264 nm Calculate the percentage of C16H19ClN2 C4H4O4 taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = = = = absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of chlorpheniramine maleate in the Sample solution (mg/mL)
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 265 nm Standard solution: USP Chlorpheniramine Maleate RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C16H19ClN2 C4H4O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS
Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride Extended-Release Capsules

(Comment on this Monograph)id=m16503=Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride ExtendedRelease Capsules=Chl-Cy-Monos.pdf) DEFINITION Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride Extended-Release Capsules contain NLT 90.0% and NMT 110.0% of the labeled amounts of chlorpheniramine maleate (C16H19ClN2 C4H4O4) and phenylpropanolamine hydrochloride (C9H13NO HCl). IDENTIFICATION A. The retention time of the major peak for chlorpheniramine maleate in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. B. The retention time of the major peak for phenylpropanolamine hydrochloride in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Phenylpropanolamine hydrochloride. ASSAY CHLORPHENIRAMINE MALEATE Mobile phase: Methanol and water (3:2) containing 0.34 g of monobasic potassium phosphate, 0.05 g of triethylamine hydrochloride, 0.025 g of sodium lauryl sulfate, and 0.1 mL of phosphoric acid in each 100 mL of solution Standard stock solution: 0.8 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: 8 g/mL of USP Chlorpheniramine Maleate RS from Standard stock solution in phosphoric acid solution (1 in 1000) Sample stock solution: Transfer NLT 10 Capsules to a suitable container, add 100 mL of water and 10 mL of phosphoric acid solution (1 in 20), and heat gently until the Capsules are fully dispersed. Cool to room temperature.
USP 32
Sample solution: 8 g/mL of chlorpheniramine maleate from Sample stock solution in water System suitability solution: Standard solution and Standard solution prepared as directed in the Assay for Phenylpropanolamine hydrochloride (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Inject 20 L of the System suitability solution and Standard solution. Suitability requirements Resolution: NLT 2.0 between phenylpropanolamine and chlorpheniramine, System suitability solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution. Calculate the percentage of chlorpheniramine maleate in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PHENYLPROPANOLAMINE HYDROCHLORIDE Mobile phase: Proceed as directed in the Assay for Chlorpheniramine maleate. Standard stock solution: 2.5 mg/mL of USP Phenylpropanolamine Hydrochloride RS in water. Standard solution: Transfer 1.0 mL of Standard stock solution to a 50-mL volumetric flask, add 5 mL of methanol, and dilute with phosphoric acid solution (1 in 1000) to volume. Sample stock solution: Transfer NLT 10 Capsules to a suitable container, add 100 mL of water and 10 mL of phosphoric acid solution (1 in 20), and heat gently until the Capsules are fully dispersed. Cool to room temperature. Sample solution: 50 g/mL of phenylpropanolamine hydrochloride solution from Sample stock solution in water. System suitability solution: Standard solution and Standard solution prepared as directed in the Assay for Chlorpheniramine maleate (1:1) Chromatographic system: Proceed as directed in the Assay for Chlorpheniramine maleate. Analysis Samples: Standard solution and Sample solution. Calculate the percentage of phenylpropanolamine hydrochloride in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = = = = peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (g/mL) rU rS CS CU = = = =

PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Times: 3, 6, and 12 h Analysis: Determine the amounts of C16H19ClN2 C4H4O4 and C9H13NO HCl dissolved as directed in the Assay for Chlorpheniramine maleate and the Assay for Phenylpropanolamine hydrochloride. Tolerances: The percentages of the labeled amounts of C16H19ClN2 C4H4O4 and C9H13NO HCl dissolved at the specified times conform to Acceptance Table 2.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS USP Phenylpropanolamine Hydrochloride RS
Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride Extended-Release Tablets

(Comment on this Monograph)id=m16504=Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride ExtendedRelease Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlorpheniramine Maleate and Phenylpropanolamine Hydrochloride Extended-Release Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of chlorpheniramine maleate (C16H19ClN2 C4H4O4) and phenylpropanolamine hydrochloride (C9H13NO HCl). IDENTIFICATION A. The retention time of the major peak for chlorpheniramine maleate in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. B. The retention time of the major peak for phenylpropanolamine hydrochloride of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Phenylpropanolamine hydrochloride. ASSAY CHLORPHENIRAMINE MALEATE Mobile phase: Methanol and water (3:2) containing 0.34 g of monobasic potassium phosphate, 0.05 g of triethylamine hydrochloride, 0.025 g of sodium lauryl sulfate, and 0.1 mL of phosphoric acid in each 100 mL of solution Standard stock solution: 0.8 mg/mL of USP Chlorpheniramine Maleate RS in water.
34

Analysis Samples: Standard solution and Sample solution Calculate the percentage of phenylpropanolamine hydrochloride in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100
USP 32
Standard solution: 8 g/mL from Standard stock solution in phosphoric acid solution (1 in 1000) Sample stock solution: Transfer the equivalent of 40 mg of chlorpheniramine maleate from finely powdered Tablets (NLT 20 Tablets), to a suitable container, add 100 mL of water and 10 mL of phosphoric acid solution (1 in 20), and heat gently until the powder is fully dispersed. Cool to room temperature. Sample solution: 8 g/mL of chlorpheniramine maleate, prepared as follows. Transfer a volume of Sample stock solution equivalent to 0.8 mg of chlorpheniramine maleate, to a 100-mL volumetric flask. Dilute with water to volume. Dilute a portion of this solution with phosphoric acid solution (1 in 1000) to obtain the Sample solution. System suitability solution: Standard solution and Standard solution prepared as directed in the Assay for Phenylpropanolamine hydrochloride (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Inject 20 L of the System suitability solution and Standard solution Suitability requirements Resolution: NLT 2.0 between phenylpropanolamine and chlorpheniramine, System suitability solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of chlorpheniramine maleate in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PHENYLPROPANOLAMINE HYDROCHLORIDE Mobile phase: Proceed as directed in the Assay for Chlorpheniramine maleate Standard stock solution: 2.5 mg/mL of USP Phenylpropanolamine Hydrochloride RS in water. Standard solution: Transfer 1.0 mL of this solution to a 50mL volumetric flask, add 5 mL of methanol, and dilute with phosphoric acid solution (1 in 1000) to volume. Sample stock solution: Transfer the equivalent of 500 mg of phenylpropanolamine hydrochloride from finely powdered Tablets (NLT 20 Tablets), to a suitable container, add 100 mL of water and 10 mL of phosphoric acid solution (1 in 20), and heat gently until the powder is fully dispersed. Cool to room temperature. Sample solution: Transfer a volume of Sample stock solution equivalent to 5 mg of phenylpropanolamine hydrochloride, to a 100-mL volumetric flask. Dilute with water to volume. System suitability solution: Standard solution and Standard solution prepared as directed in the Assay for Chlorpheniramine maleate (1:1) Chromatographic system: Proceed as directed in the Assay for Chlorpheniramine maleate. rU rS CS CU = = = =
peak response from the Sample solution peak response from the Standard solution concentration of Standard solution (g/mL) nominal concentration of Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS CU PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Times: 3, 6, and 12 h Analysis: Determine the amounts of C16H19ClN2 C4H4O4 and C9H13NO HCl dissolved as directed in the Assay for Chlorpheniramine maleate and the Assay for Phenylpropanolamine hydrochloride. Tolerances: The percentages of the labeled amounts of C16H19ClN2 C4H4O4 and C9H13NO HCl dissolved at the specified times conform to Acceptance Table 2.
= = = =
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS USP Phenylpropanolamine Hydrochloride RS
Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Extended-Release Capsules

(Comment on this Monograph)id=m16510=Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Extended-Release Capsules=Chl-Cy-Monos.pdf) DEFINITION Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Extended-Release Capsules contain NLT 90.0% and NMT 110.0% of the labeled amounts of chlorpheniramine maleate (C16H19ClN2 C4H4O4) and pseudoephedrine hydrochloride (C10H15NO HCl). IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Pseudoephedrine hydrochloride.
USP 32
ASSAY CHLORPHENIRAMINE MALEATE Mobile phase: Methanol and water (3:2) containing 0.34 g of monobasic potassium phosphate, 0.15 g of triethylamine hydrochloride, 0.25 g of sodium lauryl sulfate, and 0.1 mL of phosphoric acid in each 100 mL of solution Standard stock solution: 0.8 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: 8 g/mL of USP Chlorpheniramine Maleate RS from Standard stock solution in phosphoric acid solution (1 in 1000) Sample stock solution: Transfer NLT 10 Capsules to a suitable container, add 100 mL of water and 10 mL of phosphoric acid solution (1 in 20), and heat gently until the Capsules are fully dispersed. Cool to room temperature. Sample solution: 8 g/mL of chlorpheniramine maleate from Sample stock solution in water System suitability solution: Standard solution, and Standard solution prepared as directed in the Assay for Pseudoephedrine hydrochloride (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: NLT 2.0 between pseudoephedrine and chlorpheniramine, System suitability solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Inject 10 L of the Standard solution and the Sample solution. Calculate the percentage of chlorpheniramine maleate in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (g/mL) nominal concentration of chlorpheniramine maleate in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE HYDROCHLORIDE Mobile phase: Proceed as directed in the Assay for Chlorpheniramine maleate. Standard stock solution: 3.0 mg/mL of USP Pseudoephedrine Hydrochloride RS in water Standard solution: 0.12 mg/mL from Standard stock solution in 0.1% phosphoric acid Sample stock solution: Transfer NLT 10 Capsules to a suitable container. Add 100 mL of water and 10 mL of a phosphoric acid solution (1 in 20), and heat gently until the Capsules are fully dispersed. Cool to room temperature. Sample solution: 0.12 mg/mL of pseudoephedrine hydrochloride from Sample stock solution in water System suitability solution: Standard solution, and Standard solution prepared as directed in the Assay for Chlorpheniramine maleate (1:1) Chromatographic system: Proceed as directed in the Assay for Chlorpheniramine Maleate. Analysis Samples: Inject 10 L of the Standard solution and Sample solution. Calculate the percentage of pseudoephedrine hydrochloride in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = = = = rU rS CS CU

peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of pseudoephedrine hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 1. Medium: Water; 900 mL Apparatus 2: 50 rpm Times: 3, 6, and 12 h Analysis: Determine the amounts of C16H19ClN2 C4H4O4 and C10H15NO HCl dissolved by employing the methods set forth in the Assay for Chlorpheniramine maleate and the Assay for Pseudoephedrine hydrochloride, respectively. Tolerances: The percentages of the labeled amounts of C16H19ClN2 C4H4O4 and C10H15NO HCl dissolved at the specified times conform to Acceptance Table 2.
= = = =
Test 2: If the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2. Medium 1: Simulated gastric fluid TS, prepared without pepsin; 900 mL Medium 2: Simulated intestinal fluid TS, prepared without pancreatin; 900 mL Apparatus 2: 50 rpm Time for Medium 1: 1.5 h Times for Medium 2: 3 and 6 h Analysis: Determine the amounts of C16H19ClN2 C4H4O4 and C10H15NO HCl dissolved by using the methods set forth in the Assay for Chlorpheniramine maleate and the Assay for Pseudoephedrine hydrochloride, respectively, using Standard solutions having known concentrations of the relevant USP Reference Standard in the appropriate Medium. Tolerances: The percentages of the labeled amounts of C16H19ClN2 C4H4O4 and C10H15NO HCl dissolved at the specified times conform to Acceptance Table 2.
Time (h) 1.5 3.0 6.0 Amount Dissolved (Medium 1) 15%40% Amount Dissolved (Medium 2) 35%75% NLT 50%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. LABELING: The labeling indicates the Dissolution Test with which the product complies. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS USP Pseudoephedrine Hydrochloride RS
36

CU
USP 32
= nominal concentration of chlorpheniramine maleate in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PSEUDOEPHEDRINE HYDROCHLORIDE Mobile phase: Proceed as directed in the Assay for Chlorpheniramine maleate. Standard stock solution: 1.5 mg/mL of USP Pseudoephedrine Hydrochloride RS in water Standard solution: Transfer 1.0 mL of Standard stock solution to a 10-mL volumetric flask, add 8 mL of Mobile phase, and dilute with water to volume. Sample solution: Transfer a volume of Oral Solution, equivalent to 15 mg of pseudoephedrine hydrochloride, to a 100-mL volumetric flask. Add 80 mL of Mobile phase, dilute with water to volume, mix, and filter. System suitability solution: Standard solution for pseudoephedrine hydrochloride and Standard solution prepared as directed in the Assay for Chlorpheniramine maleate (1:1) Chromatographic system: Proceed as directed in the Assay for Chlorpheniramine maleate. Analysis Samples: Standard solution and Sample solution (10 L) Calculate the percentage of C10H15NO HCl in the portion of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Pseudoephedrine Hydrochloride RS in the Standard solution (g/mL) CU = nominal concentration of pseudoephedrine hydrochloride in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For Oral Solution packaged in single-unit containers: meets the requirements DELIVERABLE VOLUME 698: For Oral Solution packaged in multiple-unit containers: meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpheniramine Maleate RS USP Pseudoephedrine Hydrochloride RS
Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Oral Solution

(Comment on this Monograph)id=m16514=Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Oral Solution=ChlCy-Monos.pdf) DEFINITION Chlorpheniramine Maleate and Pseudoephedrine Hydrochloride Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amounts of chlorpheniramine maleate (C16H19ClN2 C4H4O4) and pseudoephedrine hydrochloride (C10H15NO HCl). IDENTIFICATION A. The retention time of the major peak for chlorpheniramine maleate in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Chlorpheniramine maleate. B. The retention time of the major peak for pseudoephedrine hydrochloride in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Pseudoephedrine hydrochloride. ASSAY CHLORPHENIRAMINE MALEATE Mobile phase: Methanol and water (3:2) containing 0.34 g of monobasic potassium phosphate, 0.15 g of triethylamine hydrochloride, 0.25 g of sodium lauryl sulfate, and 0.1 mL of phosphoric acid in each 100 mL of solution Standard stock solution: 1 mg/mL of USP Chlorpheniramine Maleate RS in water Standard solution: Transfer 1.0 mL of Standard stock solution to a 100-mL volumetric flask, add 80 mL of Mobile phase, and dilute with water to volume. This solution contains 10 g/mL. Sample solution: Equivalent to 10 g/mL of chlorpheniramine maleate from Oral Solution, in Mobile phase and water (4:1) System suitability solution: Standard solution for chlorpheniramine maleate and Standard solution prepared as directed in the Assay for Pseudoephedrine hydrochloride (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 15-cm; packing L11 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution Suitability requirements Resolution: NLT 2.0 between pseudoephedrine and chlorpheniramine, System suitability solution Tailing factor: NMT 2.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution (10 L) Calculate the percentage of C16H19CIN2 C4H4O4 in the portion of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlorpheniramine Maleate RS in the Standard solution (g/mL)
Chlorpromazine
(Comment on this Monograph)id=m16630=Chlorpromazine=Chl-Cy-Monos.pdf)
318.86 C17H19ClN2S 10H-Phenothiazine-10-propanamine, 2-chloro-N,N-dimethyl-; 2-Chloro-10-[3-(dimethylamino)propyl]phenothiazine [50-53-3].
USP 32
DEFINITION Chlorpromazine contains NLT 98.0% and NMT 101.0% of C17H19ClN2S, calculated on the dried basis. [NOTEThroughout the following analyses, protect sample or Assay specimens, the Reference Standard, and solutions containing them, by conducting the analyses without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. INFRARED ABSORPTION Sample solution: 10 mg/mL in carbon disulfide Standard solution: Dissolve 55 mg of USP Chlorpromazine Hydrochloride RS in 3 mL of 1 N sodium hydroxide, and extract the resulting solution with 5.0 mL of carbon disulfide. Acceptance criteria: The IR absorption spectrum of the Sample solution in a 1.0-mm cell between 7 and 15 m exhibits maxima only at the same wavelengths as that of the Standard solution. B. The principal spot found in the test for Procedure: Other alkylated phenothiazines corresponds in RF to the spot from the Standard stock solution. ASSAY PROCEDURE Sample: 750 mg of Chlorpromazine Analysis: Dissolve the Sample in 25 mL of glacial acetic acid in a 250-mL conical flask, warming gently on a steam bath to effect solution. Cool, add crystal violet TS, and titrate with 0.1 N perchloric acid VS to a blue endpoint. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 31.89 mg of C17H19ClN2S. Acceptance criteria: 98.0%101.0% IMPURITIES Organic Impurities PROCEDURE: OTHER ALKYLATED PHENOTHIAZINES Adsorbent: Chromatographic silica gel mixture Sample solution: 4.5 mg/mL of chlorpromazine in methanol Standard stock solution: 5 mg/mL of USP Chlorpromazine Hydrochloride RS in methanol Standard solution: 25 g/mL from Standard stock solution in methanol Application volume: 10 L Developing solvent system: Freshly prepared mixture of ether and ethyl acetate saturated with ammonium hydroxide (1:1) Analysis Samples: Sample solution, Standard stock solution, and Standard solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram, using the Developing solvent system, until the solvent front has moved 10 cm from the origin. Remove the plate from the chamber, and air-dry for 20 min. View under shortwavelength UV light. Acceptance criteria: The area and intensity of any spot, other than the principal spot, from the solution of Chlorpromazine are not greater than those of the spot from the Standard solution (0.5%). SPECIFIC TESTS LOSS ON DRYING 731: Dry in a vacuum at room temperature for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpromazine Hydrochloride RS
Official Monographs / Chlorpromazine 37
Chlorpromazine Suppositories
(Comment on this Monograph)id=m16660=Chlorpromazine Suppositories=Chl-Cy-Monos.pdf) DEFINITION Chlorpromazine Suppositories contain NLT 90.0% and NMT 110.0% of the labeled amount of C17H19ClN2S. [NOTEThroughout the following Analyses, protect Sample specimens, the Reference Standard, and solutions containing them, by conducting the Analyses without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION PROCEDURE: The principal spot found in the test for other alkylated phenothiazines corresponds in RF to the spot from the Standard stock solution. ASSAY PROCEDURE Standard solution: 5.5 g/mL of USP Chlorpromazine Hydrochloride RS in 0.1 N hydrochloric acid Sample stock solution: Place NLT 10 Suppositories in a 250-mL beaker, reduce the mass to the consistency of a paste by crushing with a spatula, and mix. Weigh a portion of the mass, equivalent to 50 mg of chlorpromazine, place in a beaker, and dissolve in 40 mL of ether. Transfer to a 250-mL separator with the aid of three 25-mL portions of ether, and extract with four 75-mL portions of 0.1 N hydrochloric acid, collecting the aqueous extracts in a 500mL volumetric flask, and add 0.1 N hydrochloric acid to volume. Sample solution: Transfer 10.0 mL of the Sample stock solution to a 200-mL volumetric flask, add 0.1 N hydrochloric acid to volume. Spectrometric conditions Mode: Spectrophotometry Analytical Wavelength: 254 nm and 277 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis: Determine the absorbances of the Standard solution and the Sample solution. Calculate the percentage of C17H19ClN2S in the portion of Suppositories taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 = (A254A277), difference of absorbances for the Sample solution = (A254A277), difference of absorbances for the AS Standard solution = concentration of the Standard solution (g/mL) CS = concentration of the Sample solution (g/mL) CU = molecular weight ratio of chlorpromazine, Mr1 318.86 = molecular weight of chlorpromazine Mr2 hydrochloride, 355.32 Acceptance criteria: 90.0%110.0% AU IMPURITIES Organic Impurities PROCEDURE: OTHER ALKYLATED PHENOTHIAZINES Adsorbent: Chromatographic silica gel mixture Sample solution: Transfer a portion of suppositories equivalent to 45 mg of chlorpromazine to a stoppered centrifuge tube, add 10 mL of methanol, shake vigorously to disperse the solid, warming gently if necessary, and centrifuge. Standard stock solution: 5 mg/mL of USP Chlorpromazine Hydrochloride RS in methanol Standard solution: 25 g/mL from Standard stock solution in methanol
38
Chlorpromazine / Official Monographs

Application volume: 10 L Developing solvent system: Freshly prepared mixture of ether and ethyl acetate saturated with ammonium hydroxide (1:1) Analysis Samples: Sample solution, Standard stock solution, and Standard solution Analysis: Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram, using the Developing solvent system, until the solvent front has moved 10 cm from the origin. Remove the plate from the chamber, and air-dry for 20 min. View under shortwavelength UV light. Acceptance criteria: The area and intensity of any spot, other than the principal spot, from the Sample solution are not greater than those of the spot from the Standard solution (0.5%).
USP 32
Application volume: 10 L Developing solvent system: Freshly prepared mixture of ether and ethyl acetate saturated with ammonium hydroxide (1:1) Analysis Samples: Standard stock solution, Standard solution, and Sample solution Proceed as directed under Chromatography 621, ThinLayer Chromatography. Develop the chromatogram, using Developing solvent system, until the solvent front has moved 10 cm from the origin. Remove the plate from the chamber, and air-dry for 20 min. View under shortwavelength UV light. Acceptance criteria: The area and intensity of any spot, other than the principal spot, from the solution of Chlorpromazine are not greater than those of the spot from the Standard solution (0.5%). SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 195198 LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpromazine Hydrochloride RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Chlorpromazine Hydrochloride RS
Chlorpromazine Hydrochloride
(Comment on this Monograph)id=m16690=Chlorpromazine Hydrochloride=Chl-Cy-Monos.pdf) C17H19ClN2S HCl 355.33 10H-Phenothiazine-10-propanamine, 2-chloro-N,N-dimethyl-, monohydrochloride; 2-Chloro-10-[3-(dimethylamino)propyl]phenothiazine monohydrochloride [69-09-0]. DEFINITION Chlorpromazine Hydrochloride contains NLT 98.0% and NMT 101.5% of C17H19ClN2S HCl, calculated on the dried basis. [NOTEThroughout the following analyses, protect sample specimens, the Reference Standard, and solutions containing them, by conducting the analyses without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. INFRARED ABSORPTION 197K B. The principal spot found in the test for Procedure: other alkylated phenothiazines corresponds in RF to the spot from the Standard stock solution. C. IDENTIFICATION TESTSGENERAL, Chloride 191: A 100mg/mL solution meets the requirements of the test. ASSAY PROCEDURE Sample: 700 mg of Chlorpromazine Hydrochloride Analysis: Dissolve the Sample in 75 mL of glacial acetic acid in a beaker and add 10 mL of mercuric acetate TS. Titrate with 0.1 N perchloric acid VS. Each mL of 0.1 N perchloric acid is equivalent to 35.53 mg of C17H19ClN2S HCl. Acceptance criteria: 98.0%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE: OTHER ALKYLATED PHENOTHIAZINES Adsorbent: Chromatographic silica gel mixture Standard stock solution: 5 mg/mL of USP Chlorpromazine Hydrochloride RS in methanol Standard solution: 25 g/mL of Standard stock solution in methanol Sample solution: 5 mg/mL in methanol (chlorpromazine hydrocloride is previously dried)
Chlorpromazine Hydrochloride Oral Concentrate

(Comment on this Monograph)id=m16692=Chlorpromazine Hydrochloride Oral Concentrate=Chl-Cy-Monos.pdf) DEFINITION Chlorpromazine Hydrochloride Oral Concentrate contains NLT 90.0% and NMT 110.0% of the labeled amount of C17H19ClN2S HCl. [NOTEThroughout the following analyses, protect sample specimens, the Reference Standard, and solutions containing them, by conducting the analyses without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 0.2 mg/mL of USP Chlorpromazine Hydrochloride RS in methanol Sample solution: Transfer a volume of Oral Concentrate, equivalent to 20 mg of chlorpromazine hydrochloride, to a 125-mL separator. Add 10 mL of water and 2 mL of sodium hydroxide solution (1 in 2). Extract with three 30-mL portions of ether. Filter the combined ether extracts through anhydrous sodium sulfate. With the aid of a stream of nitrogen, evaporate the ether to 5 mL. Quantitatively transfer the solution to a 40-mL centrifuge tube. Evaporate with a stream of nitrogen and mild heat to dryness. Dissolve the residue in 100 mL of methanol. Spray reagent: Dissolve 100 mg of platinic chloride in 10 mL of 0.1 N hydrochloric acid, add 25 mL of potassium iodide solution (1 in 25), 0.5 mL of formic acid, and dilute with water to 100 mL. Application volume: 15 L Developing solvent system: Freshly prepared mixture of ethyl acetate that has been saturated with ammonium hydroxide, ether, and methanol (15:5:4) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in the
USP 32
Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, air-dry, and spray with Spray reagent. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. CHLORIDE AND SULFATE, Chloride 191: Dilute a portion of the Oral Concentrate with an equal volume of water: this solution meets the requirements of the tests. ASSAY PROCEDURE Sample solution: Transfer the equivalent of 10 mg of chlorpromazine hydrochloride from Oral Concentrate to a 50-mL volumetric flask, and add 0.1 N hydrochloric acid to volume. Pipet 10 mL of the solution into a 250-mL separator, add 20 mL of water, render alkaline with ammonium hydroxide, and extract with four 25-mL portions of ether. Extract the combined ether extracts with four 25mL portions of 0.1 N hydrochloric acid, collecting the aqueous extracts in a 250-mL volumetric flask. Aerate to remove residual ether, and add 0.1 N hydrochloric acid to volume. Standard solution: 8 g/mL of USP Chlorpromazine Hydrochloride RS in 0.1 N hydrochloric acid Spectrometric conditions Mode: Spectrophotometry Analytical wavelengths: 254 and 277 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis: Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C17H19ClN2S HCl in each mL of the Oral Concentrate taken: Result = (AU/AS) (CS/CU) 100 = (A254 A277), difference of absorbances for the Sample solution = (A254 A277), difference of absorbances for the AS Standard solution = concentration of USP Chlorpromazine CS Hydrochloride RS in the Standard solution (g/mL) CU = nominal concentration of chlorpromazine hydrochloride in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% AU IMPURITIES Organic Impurities PROCEDURE: LIMIT OF CHLORPROMAZINE SULFOXIDE Adsorbent: 0.25-mm layer of chromatographic silica gel Chlorpromazine sulfoxide standard solution: Transfer 5 mL of 10.6 mg/mL of USP Chlorpromazine Hydrochloride RS in dilute hydrochloric acid (1 in 100) to a 50-mL volumetric flask. Add 2 mL of 30% hydrogen peroxide and heat at 60 for 10 min. Cool, and dilute with 1 M sodium bisulfite to volume. Transfer 10.0 mL to a 60-mL separator, and add 2 mL of sodium hydroxide solution (1 in 2). Extract with three 30-mL portions of ether. Filter the extracts through ether-wetted anhydrous sodium sulfate into a 250-mL conical flask. Cautiously evaporate the extracts to dryness. Dissolve the residue in 10.0 mL of methanol, and filter if necessary. Each mL of this solution contains 1 mg of chlorpromazine sulfoxide. Sample solution: Transfer the equivalent of 20 mg of chlorpromazine hydrochloride, from Oral Concentrate, to a 125-mL separator. Add 10 mL of water and 2 mL of

sodium hydroxide solution (1 in 2). Extract with three 30mL portions of ether. Filter the combined ether extracts through anhydrous sodium sulfate. With the aid of a stream of nitrogen evaporate the ether to 5 mL. Quantitatively transfer the solution to a 40-mL centrifuge tube. Evaporate with a stream of nitrogen and mild heat to dryness. Dissolve the residue in 1.0 mL of methanol. Spray reagent: Dissolve 100 mg of platinic chloride in 10 mL of 0.1 N hydrochloric acid, add 25 mL of potassium iodide solution (1 in 25), 0.5 mL of formic acid, and dilute with water to 100 mL. Application volume: 15 L Developing solvent system: Freshly prepared mixture of ethyl acetate that has been saturated with ammonium hydroxide, ether, and methanol (15:5:4) Analysis Samples: Chlorpromazine sulfoxide standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, air-dry, and spray with Spray reagent. Acceptance criteria: The chromatogram from the Sample solution may exhibit a secondary spot whose RF value corresponds to, and whose size and intensity are not greater than, those of the spot of the Chlorpromazine sulfoxide standard solution (5.0%). SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements for the absence of Escherichia coli. PH 791: 2.34.1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: Label it to indicate that it must be diluted prior to administration. USP REFERENCE STANDARDS 11 USP Chlorpromazine Hydrochloride RS
Chlorpromazine Hydrochloride Injection

(Comment on this Monograph)id=m16700=Chlorpromazine Hydrochloride Injection=Chl-Cy-Monos.pdf) DEFINITION Chlorpromazine Hydrochloride Injection is a sterile solution of Chlorpromazine Hydrochloride in Water for Injection. It contains, in each mL, NLT 23.75 mg and NMT 26.25 mg of C17H19ClN2S HCl. [NOTEThroughout the following analyses, protect sample specimens, the Reference Standard, and solutions containing them, by conducting the analyses without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 2.5 mg/mL of USP Chlorpromazine Hydrochloride RS in dilute methanol (9 in 10) Sample solution: 2.5 mg/mL of chlorpromazine hydrochloride from Injection in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
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Application volume: 10 L Developing solvent system: Freshly prepared mixture of equal volumes of ether and ethyl acetate saturated with ammonium hydroxide Analysis Samples: Standard solution and Sample solution Dry the applied solutions with the aid of a stream of nitrogen. Develop the chromatogram, using the Developing solvent system, until the solvent front has moved 13 cm from the origin. Remove the plate from the chamber, and air-dry for 30 min. Examine under short-wavelength UV light. Acceptance criteria: The area and intensity of the only other spot from the Sample solution, other than the principal spot, are not greater than those of the spot from the Standard solution (5.0%). SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: It contains NMT 6.9 USP Endotoxin Units/mg of chlorpromazine hydrochloride. PH 791: 3.45.4 INJECTIONS 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Chlorpromazine Hydrochloride RS USP Endotoxin RS
Mode: TLC Adsorbent: Chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Freshly prepared mixture of equal volumes of ether and ethyl acetate saturated with ammonium hydroxide Analysis Samples: Standard solution and Sample solution Develop the chromatogram in the Developing solvent system until the solvent front has moved 10 cm from the origin. Remove the plate from the developing chamber, air-dry for 20 min, then view under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. IDENTIFICATION TESTSGENERAL, Chloride 191 It meets the requirements. ASSAY PROCEDURE Sample solution: Transfer the equivalent of 100 mg of chlorpromazine hydrochloride from Injection to a 500-mL volumetric flask, and add 0.1 N hydrochloric acid to volume. Pipet 10 mL of the solution into a 250-mL separator, add 20 mL of water, render alkaline with ammonium hydroxide, and extract with four 25-mL portions of ether. Extract the combined ether extracts with four 25-mL portions of 0.1 N hydrochloric acid, and collect the aqueous extracts in a 250mL volumetric flask. Aerate to remove residual ether, and add 0.1 N hydrochloric acid to volume. Standard solution: 8 g/mL of USP Chlorpromazine Hydrochloride RS in 0.1 N hydrochloric acid Spectrometric conditions Mode: Spectrophotometry Analytical wavelengths: 254 and 277 nm Cell: 1cm Blank: 0.1 N hydrochloric acid Analysis Samples: Sample solution and Standard solution Determine the absorbances of the Standard solution and Sample solution at the wavelength of maximum absorbance. Calculate the quantity, in mg, of C17H19ClN2S HCl in each mL of the Injection taken: Result = (12.5 CS AU)/(AS V) = concentration of USP Chlorpromazine Hydrochloride RS in the Standard solution (g/mL) = (A254 A277), difference in the absorbances for AU the Sample solution = (A254 A277), difference in the absorbances for AS the Standard solution V = volume of Injection taken (mL) Acceptance criteria: 23.7526.25 mg/mL CS IMPURITIES Organic Impurities PROCEDURE: LIMIT OF CHLORPROMAZINE SULFOXIDE [NOTEConduct this test without exposure to daylight, and with the minimum necessary exposure to artificial light.] Standard solution: 50 g/mL of USP Chlorpromazine Hydrochloride RS in methanol Sample solution: Sample solution prepared as directed in Identification test A and methanol (2:3) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture
Chlorpromazine Hydrochloride Syrup

(Comment on this Monograph)id=m16720=Chlorpromazine Hydrochloride Syrup=Chl-Cy-Monos.pdf) DEFINITION Chlorpromazine Hydrochloride Syrup contains, in each 100 mL, NLT 190 and NMT 210 mg of C17H19ClN2S HCl. [NOTEThroughout the following Analyses, protect sample specimens, the Reference Standard, and solutions containing them, by conducting the Analyses without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel Standard solution: 0.2 mg/mL of USP Chlorpromazine Hydrochloride RS in methanol Sample solution: Transfer a volume of Syrup, equivalent to 20 mg of chlorpromazine hydrochloride, to a 125-mL separator. Add 10 mL of water, 2 mL of sodium hydroxide solution (1 in 2), and mix. Extract with three 30-mL portions of ether. Filter the combined ether extracts through anhydrous sodium sulfate. With the aid of a stream of nitrogen, evaporate the ether to 5 mL. Quantitatively transfer the solution to a 40-mL centrifuge tube. Evaporate with a stream of nitrogen and mild heat to dryness. Dissolve the residue in 100 mL of methanol. Spray reagent: Dissolve 100 mg of platinic chloride in 10 mL of 0.1 N hydrochloric acid, add 25 mL of potassium iodide solution (1 in 25), 0.5 mL of formic acid, and dilute with water to 100 mL. Application volume: 15 L Developing solvent system: Freshly prepared mixture of ethyl acetate that has been saturated with ammonium hydroxide, ether, and methanol (15:5:4)
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Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, air-dry, and spray with Spray reagent. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. IDENTIFICATION TESTSGENERAL, Chloride 191: Dilute a portion of the Syrup with an equal volume of water. This solution meets the requirements. ASSAY PROCEDURE Standard solution: 8 g/mL of USP Chlorpromazine Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: Transfer the equivalent of 10 mg of chlorpromazine hydrochloride from Syrup, to a 50-mL volumetric flask, and add 0.1 N hydrochloric acid to volume. Pipet 10 mL of the solution into a 250-mL separator, add 20 mL of water, render alkaline with ammonium hydroxide, and extract with four 25-mL portions of ether. Extract the combined ether extracts with four 25-mL portions of 0.1 N hydrochloric acid, collecting the aqueous extracts in a 250mL volumetric flask. Aerate to remove residual ether, and add 0.1 N hydrochloric acid to volume. Spectrometric conditions Mode: Spectrophotometry Analytical wavelengths: 254 and 277 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis: Determine the absorbances of the Standard solution and Sample solution at the wavelength of maximum absorbance. Calculate the quantity, in mg, of C17H19ClN2S HCl in each mL of the Syrup taken: Result = (AUw254AUw277)/(ASw254ASw277) (CS/CU) F AUw254 = absorbance of the Sample solution at wavelength 254 AUw277 = absorbance of the Sample solution at wavelength 277 ASw254 = absorbance of the Standard solution at wavelength 254 ASw277 = absorbance of the Standard solution at wavelength 277 CS = concentration of USP Chlorpromazine Hydrochloride RS in the Standard solution (g/mL) CU = nominal concentration of chlorpromazine hydrochloride in the Sample solution (g/mL) F = conversion factor (g to mg) Acceptance criteria: 190210 mg/100 mL IMPURITIES Organic Impurities PROCEDURE: LIMIT OF CHLORPROMAZINE SULFOXIDE Adsorbent: 0.25-mm layer of chromatographic silica gel Chlorpromazine sulfoxide standard solution: Transfer 5 mL of 10.6 mg/mL of USP Chlorpromazine Hydrochloride RS in dilute hydrochloric acid (1 in 100) to a 50-mL volumetric flask. Add 2 mL of 30% hydrogen peroxide, and heat at 60 for 10 min. Cool, and dilute with 1 M sodium bisulfite to volume. Transfer 10.0 mL to a 60-mL separator, add 2 mL of sodium hydroxide solution (1 in 2), and mix. Extract with three 30-mL portions of ether. Filter the extracts through ether-wetted anhydrous sodium sulfate into a 250-mL conical flask. Cautiously evaporate the

extracts to dryness. Dissolve the residue in 10.0 mL of methanol, and filter, if necessary. Each mL of this solution contains 1 mg of chlorpromazine sulfoxide. Sample solution: Transfer the equivalent of 20 mg of chlorpromazine hydrochloride from Syrup to a 125-mL separator. Add 10 mL of water and 2 mL of sodium hydroxide solution (1 in 2), and mix. Extract with three 30mL portions of ether. Filter the combined ether extracts through anhydrous sodium sulfate. With the aid of a stream of nitrogen, evaporate the ether to 5 mL. Quantitatively transfer the solution to a 40-mL centrifuge tube. Evaporate with a stream of nitrogen, and mild heat to dryness. Dissolve the residue in 1.0 mL of methanol. Spray reagent: Dissolve 100 mg of platinic chloride in 10 mL of 0.1 N hydrochloric acid, add 25 mL of potassium iodide solution (1 in 25), 0.5 mL of formic acid, and dilute with water to 100 mL. Application volume: 15 L Developing solvent system: Freshly prepared mixture of ethyl acetate that has been saturated with ammonium hydroxide, ether, and methanol (15:5:4) Analysis Samples: Chlorpromazine sulfoxide standard solution and Sample solution Analysis: Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, air-dry, and spray with Spray reagent. Acceptance criteria: The chromatogram from the Sample solution may exhibit a secondary spot whose RF value corresponds to, and whose size and intensity are not greater than, those of the spot of the Chlorpromazine sulfoxide standard solution (5.0%). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlorpromazine Hydrochloride RS
Chlorpromazine Hydrochloride Tablets

(Comment on this Monograph)id=m16730=Chlorpromazine Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlorpromazine Hydrochloride Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of C17H19ClN2S HCl. [NOTEThroughout the following analyses, protect sample specimens, the Reference Standard, and solutions containing them, by conducting the procedures without delay, under subdued light, or using low-actinic glassware.] IDENTIFICATION A. The principal spot found in the test under Procedure: Other alkylated phenothiazines corresponds in RF to the spot of the Standard stock solution. B. IDENTIFICATION TESTSGENERAL, Chloride 191: Equivalent to 1 mg/mL of chlorpromazine hydrochloride from powdered Tablets in water ASSAY PROCEDURE Standard solution: 8 g/mL of USP Chlorpromazine Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: Transfer an amount equivalent to 100 mg of chlorpromazine hydrochloride from finely powdered Tablets (NLT 20 Tablets) to a 500-mL volumetric flask. Add 200 mL of water and 5 mL of hydrochloric acid, insert the
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Develop the chromatogram, using the Developing solvent system, until the solvent front has moved about 10 cm from the origin. Remove the plate from the chamber, and air-dry for 20 min. View under short-wavelength UV light. Acceptance criteria: The area and intensity of any spot, other than the principal spot, from the solution from the Tablets is not greater than that of the spot of the Standard solution (0.5%). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Chlorpromazine Hydrochloride RS
stopper, and shake for about 10 min. Dilute with water to volume, and mix. Filter a portion of the solution, discarding the first 50 mL of the filtrate. Pipet 10.0 mL of the solution into a 250-mL separator, add 20 mL of water, render alkaline with ammonium hydroxide, and extract with four 25-mL portions of ether. Extract the combined ether extracts with four 25-mL portions of 0.1 N hydrochloric acid, collecting the aqueous extracts in a 250-mL volumetric flask. Aerate to remove residual ether, and dilute with 0.1 N hydrochloric acid to volume. Spectrometric conditions Mode: Spectrophotometry Analytical wavelengths: 254 and 277 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis: Determine the absorbances of the Standard solution and Sample solution. Calculate the percentage of C17H19ClN2S HCl in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 = (A254 A277), difference in the absorbances for the Sample solution = (A254 A277), difference in the absorbances for the AS Standard solution = concentration of USP Chlorpromazine CS Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of chlorpromazine CU hydrochloride in the Sample solution (g/mL) Acceptance criteria: 95.0%105.0% AU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1: 50 rpm Time: 30 min Analytical wavelength: UV 254 nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Chlorpromazine Hydrochloride RS in Medium Tolerances: NLT 80% (Q) of the labeled amount of C17H19ClN2S HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE: OTHER ALKYLATED PHENOTHIAZINES Standard stock solution: 5 mg/mL of USP Chlorpromazine Hydrochloride RS in methanol Standard solution: 25 g/mL from Standard stock solution in methanol Sample solution: Transfer an amount equivalent to 50 mg of chlorpromazine hydrochloride from finely powdered Tablets to a stoppered centrifuge tube. Add 10 mL of methanol, shake vigorously, and centrifuge (remove any sugar coating by prior washing with water). Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: Chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Freshly prepared mixture of ether and ethyl acetate saturated with ammonium hydroxide (1:1) Analysis Samples: Standard stock solution, Standard solution, and Sample solution
Chlorpropamide
(Comment on this Monograph)id=m16780=Chlorpropamide=Chl-Cy-Monos.pdf)
276.74 C10H13ClN2O3S Benzenesulfonamide, 4-chloro-N-[(propylamino)carbonyl]-; 1-[(p-Chlorophenyl)sulfonyl]-3-propylurea [94-20-2]. DEFINITION Chlorpropamide contains NLT 97.0% and NMT 103.0% of C10H13ClN2O3S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 1 mg/mL of Chlorpropamide in acetone Developing solvent system: Methylene chloride, methanol, cyclohexane, and ammonium hydroxide (100:50:30:10) ASSAY PROCEDURE Mobile phase: Mix equal volumes of acetonitrile and dilute glacial acetic acid (1 in 100). [NOTEDo not exceed 50% of acetonitrile.] Standard solution: 0.05 mg/mL of USP Chlorpropamide RS in Mobile phase Sample solution: 0.05 mg/mL of Chlorpropamide in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 for the analyte peak Relative standard deviation: NMT 2.0% for replicate injections
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Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H13ClN2O3S in the portion of Chlorpropamide taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlorpropamide RS in the Standard solution (mg/mL) = concentration of Chlorpropamide in the Sample CU solution (mg/mL) Acceptance criteria: 97.0%103.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.4% SELENIUM 291: NMT 30 ppm HEAVY METALS, Method II 231: NMT 30 ppm SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: Between 126 and 129 LOSS ON DRYING 731: Dry a sample in a vacuum at 60 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chlorpropamide RS
Official Monographs / Chlortetracycline 43

Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H13ClN2O3S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Chlorpropamide RS in the Standard solution (mg/mL) = nominal concentration of chlorpropamide in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 60 min Detector: UV 230 nm Sample solution: Sample per Dissolution 711. Dilute with 0.1 N hydrochloric acid to a concentration that is similar to that of the Standard solution, and filter. Standard solution: USP Chlorpropamide RS in 0.1 N hydrochloric acid. [NOTEA volume of alcohol not exceeding 10% of the final volume of the Standard solution may be used to dissolve the USP Chlorpropamide RS.] Tolerances: NLT 75% (Q) of the labeled amount of C10H13ClN2O3S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chlorpropamide RS rU rS CS
Chlorpropamide Tablets
(Comment on this Monograph)id=m16810=Chlorpropamide Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlorpropamide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C10H13ClN2O3S. IDENTIFICATION [NOTEThe Sample is prepared as follows.] Sample: Shake the equivalent of 100 mg of chlorpropamide from finely powdered Tablets with 20 mL of 1 N hydrochloric acid, and extract with 50 mL of chloroform. Filter the chloroform through chloroform-washed cotton into a suitable beaker, and evaporate the chloroform on a steam bath with the aid of a current of dry air to dryness. Dry the residue at 105 for 1 h. A. INFRARED ABSORPTION 197K B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Prepare a 1 mg/mL solution of chlorpropamide from the dried Sample in acetone. Developing solvent system: Methylene chloride, methanol, cyclohexane, and ammonium hydroxide (100:50:30:10) ASSAY PROCEDURE Mobile phase: Mix equal volumes of acetonitrile and dilute glacial acetic acid (1 in 100). [NOTEDo not exceed 50% of acetonitrile.] Standard solution: 0.05 mg/mL of USP Chlorpropamide RS in the Mobile phase Sample stock solution: Transfer an amount of finely powdered Tablets (powder NLT 20 Tablets) equivalent to 50 mg of chlorpropamide to a 100-mL volumetric flask. Add Mobile phase to volume and filter, discarding the first 10 mL of the filtrate. Sample solution: Pipet 10 mL of the Sample stock solution into a second 100-mL volumetric flask, and add Mobile phase to volume.
Chlortetracycline Bisulfate
(Comment on this Monograph)id=m16930=Chlortetracycline Bisulfate=Chl-Cy-Monos.pdf) DEFINITION Chlortetracycline Bisulfate has a potency equivalent to NLT 760 g of C22H23ClN2O8 HCl per mg, calculated on the dried and butyl alcohol-free basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Wavelength: 368 nm Medium: 0.1 N hydrochloric acid Sample solution: 40 g/mL Acceptance criteria: Absorptivities, calculated on the dried and butyl alcohol-free basis, are NLT 83.0% and NMT 95.0% of that of the USP Chlortetracycline Hydrochloride RS, the potency of the Reference Standard being taken into account.
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solution from Standard solution 1. Calculate the percentage of butyl alcohol in the Sample taken: Result = (AU/AS) (WS/WU) 100 = absorbance of the Sample solution = absorbance of Standard solution 2 = weight of butyl alcohol taken to prepare Standard solution 1 (g) = weight of the Sample (g) WU Acceptance criteria: NMT 15.0% AU AS WS
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ASSAY PROCEDURE Analysis: Proceed with Chlortetracycline Bisulfate as directed for chlortetracycline under AntibioticsMicrobial Assays 81. Acceptance criteria: NLT 760 g/mg OTHER COMPONENTS SULFATE CONTENT Sample: 1 g Analysis: Transfer to a 250-mL beaker, and dissolve in 100 mL of water. Neutralize the solution with 7.5 N ammonium hydroxide to litmus paper, and warm. Filter, and wash the filter with warm water. Neutralize the filtrate with 6 N hydrochloric acid to litmus, and add an additional 4 mL of 6 N hydrochloric acid. Heat the solution to boiling, and add, with constant stirring, sufficient boiling barium chloride TS to precipitate all of the sulfate. Add an additional 2 mL of barium chloride TS, and digest on a steam bath for 1 h. Pass the mixture through ashless filter paper, transferring the residue quantitatively to the filter, and wash the residue with hot water until no precipitate is obtained when 1 mL of silver nitrate TS is added to 5 mL of washing. Transfer the paper containing the residue to a tared crucible. Char the paper, without burning, and ignite the crucible and its contents to constant weight. Perform a blank determination concurrently with the Sample determination, and subtract the weight of residue from that of the Sample determination to obtain the weight of residue attributable to the sulfate content of the specimen. Acceptance criteria: NLT 15.0%, calculated on the dried and butyl alcohol-free basis IMPURITIES Organic Impurities PROCEDURE: BUTYL ALCOHOL Solution A: 0.2 g/mL of ceric ammonium nitrate in 4 N nitric acid Standard solution 1: 3 mg/mL of butyl alcohol in water Standard solution 2: 10 mL of Standard solution 1 and 1 drop of dimethicone to a 50-mL distilling flask equipped with a condenser and an extension that reaches into a collecting tube maintained in an ice-water bath. Distill slowly, and collect about 8 mL of distillate. Warm the distillate to ambient temperature, and transfer with the aid of water to a 10-mL volumetric flask. Dilute with water to volume. Sample solution: Transfer Chlortetracycline Bisulfate, equivalent to 30 mg of butyl alcohol, to a 50-mL distilling flask equipped with a condenser and an extension that reaches into a collecting tube maintained in an ice bath. Add 25 mL of water and 1 drop of dimethicone to the distilling flask. Distill slowly, and collect 8 mL of the distillate. Warm the distillate to ambient temperature, and transfer with the aid of water to a 10-mL volumetric flask. Dilute with water to volume. Blank: Water Spectrometric conditions Mode: Visible spectrophotometry Analytical wavelength: 475 nm Analysis Samples: Standard solution 1, Standard solution 2, Sample solution, and Blank To four separate test tubes add, respectively, 5.0 mL of Standard solution 1, 5.0 mL of Standard solution 2, 5.0 mL of Sample solution, and 5.0 mL of water to provide a blank. To each add 2.0 mL of Solution A. Concomitantly determine the absorbances of the solutions from the Standard solutions and the Sample solution at the wavelength of maximum absorbance. In a suitable determination, the absorbance of the solution from Standard solution 2 is NLT 98.0% of the absorbance of the
SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements LOSS ON DRYING 731: Dry it in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: Label it to indicate that it is intended for veterinary use only. USP REFERENCE STANDARDS 11 USP Chlortetracycline Hydrochloride RS
Chlortetracycline and Sulfamethazine Bisulfates Soluble Powder

(Comment on this Monograph)id=m16936=Chlortetracycline and Sulfamethazine Bisulfates Soluble Powder=Chl-CyMonos.pdf) DEFINITION Chlortetracycline and Sulfamethazine Bisulfates Soluble Powder is a dry mixture of Chlortetracycline Bisulfate and Sulfamethazine Bisulfate and one or more suitable buffers and diluents. It contains the equivalent of NLT 85.0% and NMT 125.0% of the labeled amounts of chlortetracycline hydrochloride (C22H24N2O8 HCl) and sulfamethazine (C12H14N4O2S). ASSAY CHLORTETRACYCLINE HYDROCHLORIDE Analysis: Proceed as directed for chlortetracycline under AntibioticsMicrobial Assays 81, using an equivalent to 100 mg of chlortetracycline hydrochloride from Powder, dissolved in an accurately measured volume of 0.01 N hydrochloric acid to obtain a stock solution having a convenient concentration. Dilute an accurately measured volume of this stock solution quantitatively and stepwise with water to obtain a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 85.0%125.0% SULFAMETHAZINE Analysis: Proceed as directed under Nitrite Titration 451, using an equivalent to 500 mg of sulfamethazine from Powder. Each mL of 0.1 M sodium nitrite is equivalent to 27.83 mg of sulfamethazine. Acceptance criteria: 85.0%125.0% SPECIFIC TESTS LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers.
USP 32
LABELING: Label it to indicate that it is intended for veterinary use only. USP REFERENCE STANDARDS 11 USP Chlortetracycline Hydrochloride RS
Official Monographs / Chlortetracycline 45

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: Where it is intended for use in preparing sterile dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of sterile dosage forms. USP REFERENCE STANDARDS 11 USP Chlortetracycline Hydrochloride RS USP Oxytetracycline RS USP Tetracycline Hydrochloride RS
Chlortetracycline Hydrochloride
(Comment on this Monograph)id=m16940=Chlortetracycline Hydrochloride=Chl-Cy-Monos.pdf)
Chlortetracycline Hydrochloride Ointment

C22H23ClN2O8 HCl 515.34 2-Naphthacenecarboxamide, 7-chloro-4(dimethylamino)-1,4,4a,5,5a,6,11,12aoctahydro-3,6,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-, monohydrochloride [4S-(4,4a,5a,6,12a)]-; 7-Chloro-4-(dimethylamino)-1,4,4a,5,5a,6,11,12aoctahydro-3,6,10,12,12a-pentahydroxy-6-methyl-1,11dioxo-2-naphthacenecarboxamide monohydrochloride [64-72-2]. DEFINITION Chlortetracycline Hydrochloride has a potency of NLT 900 g of C22H23ClN2O8 HCl per mg. [NOTEChlortetracycline Hydrochloride labeled solely for use in preparing oral veterinary dosage forms has a potency of NLT 820 g of C22H23ClN2O8 HCl per mg.] IDENTIFICATION A. IDENTIFICATIONTETRACYCLINES, Method II 193: Meets the requirements Sample solution: 0.5 mg/mL in methanol System suitability solution: 0.5 mg/mL each of USP Chlortetracycline Hydrochloride RS, USP Oxytetracycline RS, and USP Tetracycline Hydrochloride RS in methanol B. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements Sample solution: 50 mg/mL ASSAY PROCEDURE Analysis: Proceed as directed under AntibioticsMicrobial Assays 81. Acceptance criteria: NLT 900 g/mg of C22H23ClN2O8 HCl; when labeled solely for use in preparing oral veterinary dosage forms: NLT 820 g/mg SPECIFIC TESTS OPTICAL ROTATION 781S: 235 to 250 Sample solution: 5 mg/mL that has been allowed to stand in the dark for 30 min CRYSTALLINITY 695: Meets the requirements PH 791: 2.33.3, in 10 mg/mL solution LOSS ON DRYING 731: NMT 2.0% Sample: 100 mg Analysis: Dry in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h. STERILITY TESTS 71: Where the label states that Chlortetracycline Hydrochloride is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration, 6 g of specimen aseptically dissolved in 200 mL of Fluid D being used. (Comment on this Monograph)id=m16959=Chlortetracycline Hydrochloride Ointment=Chl-Cy-Monos.pdf) DEFINITION Chlortetracycline Hydrochloride Ointment contains NLT 90.0% and NMT 125.0% of the labeled amount of C22H23ClN2O8 HCl in a suitable ointment base. ASSAY PROCEDURE Analysis: Proceed as directed under AntibioticsMicrobial Assays 81, using an equivalent to 30 mg of chlortetracycline hydrochloride from Ointment, shaken in a separator with about 50 mL of ether, and extracted with four 20-mL portions of 0.01 N hydrochloric acid. Combine the aqueous extracts in a 100-mL volumetric flask, and dilute with 0.01 N hydrochloric acid to volume. Dilute this stock solution quantitatively and stepwise with water to obtain a Sample Dilution having a concentration assumed to be equal to the medium dose level of the Standard. Acceptance criteria: 90.0%125.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 0.5%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in well-closed, light-resistant containers. USP REFERENCE STANDARDS 11 USP Chlortetracycline Hydrochloride RS
Chlortetracycline Hydrochloride Ophthalmic Ointment

(Comment on this Monograph)id=m16960=Chlortetracycline Hydrochloride Ophthalmic Ointment=Chl-Cy-Monos.pdf) DEFINITION Chlortetracycline Hydrochloride Ophthalmic Ointment contains NLT 90.0% and NMT 125.0% of the labeled amount of C22H23ClN2O8 HCl. ASSAY PROCEDURE: Proceed as directed under AntibioticsMicrobial Assays 81, using an equivalent to 10 mg of chlortetracycline hydrochloride from Ointment, shaken in a separator with 50 mL of ether, and extracted with four 20-
46
Chlortetracycline / Official Monographs
USP 32
LABELING: Label it to indicate that it is intended for oral veterinary use only. USP REFERENCE STANDARDS 11 USP Chlortetracycline Hydrochloride RS
mL portions of 0.01 N hydrochloric acid. Combine the aqueous extracts in a 100-mL volumetric flask, and dilute with 0.01 N hydrochloric acid to volume. Dilute this stock solution quantitatively and stepwise with water to obtain a sample dilution having a concentration assumed to be equal to the median dose level of the standard. Acceptance criteria: 90.0%125.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements WATER DETERMINATION, Method I 921: NMT 0.5%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel METAL PARTICLES IN OPHTHALMIC OINTMENTS 751: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Chlortetracycline Hydrochloride RS
Chlortetracycline Hydrochloride Tablets

(Comment on this Monograph)id=m16973=Chlortetracycline Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlortetracycline Hydrochloride Tablets contain NLT 90.0% and NMT 120.0% of the labeled amount of C22H23ClN2O8 HCl. IDENTIFICATION IDENTIFICATIONTETRACYCLINES, Method II 193 Sample solution: 0.5 mg/mL of chlortetracycline hydrochloride in methanol from an appropriately diluted quantity of finely ground Tablet powder; shake the solution, then filter Standard solution: 0.5 mg/mL each of USP Chlortetracycline Hydrochloride RS, USP Oxytetracycline RS, and USP Tetracycline Hydrochloride RS in methanol. Analysis: Proceed as directed in the chapter. ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 Sample solution: Transfer NLT 5 Tablets to a high-speed glass blender jar containing a measured volume of 0.01 N hydrochloric acid, so that after blending for 35 min a solution of convenient concentration is obtained. Analysis: Proceed as directed for chlortetracycline, using a measured volume of the Sample solution diluted with water to obtain a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISINTEGRATION 701: 1 h, simulated gastric fluid TS being used as the sample medium in place of water UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Weight Variation. SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 3.0%, or where the Tablets have a diameter of greater than 15 mm, NMT 6.0%, a quantity of finely ground Tablet powder being used ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. LABELING: Label the Tablets to indicate that they are intended for veterinary use only. USP REFERENCE STANDARDS 11 USP Chlortetracycline Hydrochloride RS USP Oxytetracycline RS USP Tetracycline Hydrochloride RS
Chlortetracycline Hydrochloride Soluble Powder

(Comment on this Monograph)id=m16963=Chlortetracycline Hydrochloride Soluble Powder=Chl-Cy-Monos.pdf) DEFINITION Chlortetracycline Hydrochloride Soluble Powder contains NLT 90.0% and NMT 125.0% of the labeled amount of C22H23ClN2O8 HCl. ASSAY PROCEDURE Sample solution 1 (where it is labeled on a weight basis): Dissolve 3 g of Powder in a measured volume of 0.01 N hydrochloric acid sufficient to obtain a solution containing NLT 1000 g/mL of C22H23ClN2O8 HCl. Sample solution 2 (where the label states the amount of chlortetracycline in the immediate container): Transfer the contents of 1 container of Powder to a measured volume of 0.01 N hydrochloric acid sufficient to obtain a solution containing NLT 1000 g/mL of C22H23ClN2O8 HCl. Analysis: Proceed with Powder as directed for chlortetracycline under AntibioticsMicrobial Assays 81, using a measured volume of Sample solution diluted quantitatively and stepwise to yield a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%125.0% SPECIFIC TESTS LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light.
USP 32
Official Monographs / Chlorthalidone 47

Tailing factor: NMT 2.0 for chlorthalidone and chlorthalidone related compound A Relative standard deviation: NMT 2.0% from five replicate injections Analysis Samples: Standard solution and Sample solution. Calculate the percentage of C14H11ClN2O4S in the portion of Chlorthalidone taken: Result = (RU/RS) (CS/CU) 100
Chlorthalidone
(Comment on this Monograph)id=m17010=Chlorthalidone=Chl-Cy-Monos.pdf)
C14H11ClN2O4S 338.77 Benzenesulfonamide, 2-chloro-5-(2,3-dihydro-1-hydroxy-3oxo-1H-isoindol-1-yl)-; 2-Chloro-5-(1-hydroxy-3-oxo-1isoindolinyl)benzenesulfonamide [77-36-1]. DEFINITION Chlorthalidone contains NLT 98.0% and NMT 102.0% of C14H11ClN2O4S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 275 nm Medium: 2 N hydrochloric acid in methanol (1 in 50) Solution: 100 g/mL Acceptance criteria: Absorptivities do not differ by more than 4.0% C. Dissolve 50 mg in 3 mL of sulfuric acid: an intense yellow color develops. ASSAY PROCEDURE Mobile phase: Methanol and 0.01 M dibasic ammonium phosphate (2:3), adjusted dropwise with phosphoric acid to a pH of 5.5 0.1 Internal standard solution: 1 mg/mL of 2,7naphthalenediol in methanol Chlorthalidone related compound A solution: 5 g/mL of USP Chlorthalidone Related Compound A RS in methanol Standard stock solution: 1 mg/mL of USP Chlorthalidone RS in methanol. Standard solution: Pipet 5 mL of Standard stock solution into a 50-mL volumetric flask containing 5.0 mL of Internal standard solution and 10.0 mL of Chlorthalidone related compound A solution, and dilute with water to volume. This solution contains 0.1 mg/mL of chlorthalidone and 1 g/mL of chlorthalidone related compound A. Sample stock solution: 1 mg/mL of Chlorthalidone in methanol. Sample solution: Pipet 5 mL of Sample stock solution into a 50-mL volumetric flask containing 5.0 mL of Internal standard solution and 10.0 mL of methanol, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements [NOTEThe relative retention times for chlorthalidone related compound A, chlorthalidone, and the internal standard are about 0.5, 0.8, and 1.0, respectively.] Resolution: NLT 1.5 between chlorthalidone and chlorthalidone related compound A, and between chlorthalidone and the internal standard
= peak response ratio of chlorthalidone to the internal standard from the Sample solution RS = peak response ratio of chlorthalidone to the internal standard from the Standard solution CS = concentration of the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% RU IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221: Shake 1.0 g with 40 mL of water for 5 min, and pass through chloride-free filter paper previously rinsed with water: the filtrate shows no more chloride than corresponds to 0.50 mL of 0.020 N hydrochloric acid (0.035%). HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE: LIMIT OF CHLORTHALIDONE RELATED COMPOUND A Analysis: Proceed as directed in the Assay, except to calculate the percentage of chlorthalidone related compound A in the portion of Chlorthalidone taken: Result = (RU/RS) (Cr/CT) (0.1) = peak response ratio of chlorthalidone related compound A to the internal standard from the Sample solution = peak response ratio of chlorthalidone related RS compound A to the internal standard from the Standard solution = concentration of USP Chlorthalidone Related Cr Compound A RS in the Standard solution (g/mL) = concentration of Chlorthalidone in the Sample CT solution (mg/mL) Acceptance criteria: NMT 1.0% [NOTEUSP Chlorthalidone Related Compound A RS is 4chloro-3-sulfamoyl-2-benzophenone carboxylic acid (CCA).] RU SPECIFIC TESTS LOSS ON DRYING 731: 0.4% of its weight. Dry 2 g at 105 for 4 h: it loses NMT
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chlorthalidone RS USP Chlorthalidone Related Compound A RS
Chlorthalidone Tablets
(Comment on this Monograph)id=m17040=Chlorthalidone Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlorthalidone Tablets contain NLT 92.0% and NMT 108.0% of the labeled amount of C14H11ClN2O4S.
48
Chlorthalidone / Official Monographs

RS
USP 32
= peak response ratio of chlorthalidone to the internal standard from the Standard solution = concentration of the Standard solution (mg/mL) CS = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 92.0%108.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 60 min Detector: UV 275 nm Sample solution: Sample per Dissolution 711. Standard solution: 5 mg/mL of USP Chlorthalidone RS in methanol Analysis: Determine the amount of C14H11ClN2O4S dissolved from UV absorbances of filtered portions of the solution under test, suitably diluted with water, in comparison with a quantitative dilution in water of the Standard solution having a known concentration of USP Chlorthalidone RS comparable to the concentration of the solution under test. Tolerances: NLT 70% (Q) of the labeled amount of C14H11ClN2O4S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chlorthalidone RS
IDENTIFICATION A. INFRARED ABSORPTION 197M Sample: Digest a quantity of powdered Tablets, equivalent to 100 mg of chlorthalidone, with 10 mL of acetone on a steam bath for about 5 min. Filter the solution into a 50-mL beaker, add 20 mL of water, and boil on the steam bath for about 5 min, passing a gentle current of air above the solution to remove the acetone. Cool in an ice bath, filter, and dry the crystals at 105 for 4 h. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol and 0.01 M dibasic ammonium phosphate (2:3), adjusted dropwise with phosphoric acid to a pH of 5.5 0.1 Internal standard solution: 1.0 mg/mL of 2,7naphthalenediol in methanol Standard stock solution: 1 mg/mL of USP Chlorthalidone RS in methanol. Standard solution: Pipet 5 mL of this solution into a 50-mL volumetric flask containing 5.0 mL of Internal standard solution and 10.0 mL of methanol, and dilute with water to volume. Sample stock solution: Transfer the equivalent to 100 mg of chlorthalidone from powdered Tablets (NLT 20), to a 100-mL volumetric flask. Dissolve in 50 mL of methanol, shake for 30 min, and dilute with methanol to volume. Transfer 30 mL of this solution to a 50-mL centrifuge tube, and centrifuge for 10 min. Sample solution: Pipet 5 mL of the clear supernatant into a 50-mL volumetric flask containing 5.0 mL of Internal standard solution and 10.0 mL of methanol. Dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for chlorthalidone and the internal standard are about 0.8 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between chlorthalidone and the internal standard Tailing factor: NMT 2.0 for the chlorthalidone and internal standard peaks Relative standard deviation: NMT 2.0% from five replicate injections Analysis Samples: Standard solution and Sample solution. Calculate the percentage of C14H11ClN2O4S in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 RU = peak response ratio of chlorthalidone to the internal standard from the Sample solution
Chlorzoxazone
(Comment on this Monograph)id=m17085=Chlorzoxazone=Chl-Cy-Monos.pdf)
C7H4ClNO2 2(3H)-Benzoxazolone, 5-chloro-; 5-Chloro-2-benzoxazolinone [95-25-0]. DEFINITION Chlorzoxazone contains NLT 98.0% and NMT 102.0% of C7H4ClNO2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Previously dried B. ULTRAVIOLET ABSORPTION 197U Medium: Methanol Sample solution: 20 g/mL ASSAY PROCEDURE Standard solution: methanol
169.56
20 g/mL of USP Chlorzoxazone RS in
USP 32
Sample solution: 20 g/mL of Chlorzoxazone in methanol Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 282 nm Cell: 1 cm Blank: Methanol Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H4ClNO2 in the portion taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Chlorzoxazone RS in the Standard solution (g/mL) CU = concentration of the Sample solution (g/mL) Acceptance criteria: 98.0%102.0% AU AS CS OTHER COMPONENTS CHLORINE CONTENT Sample solution: Dissolve 300 mg in 10 mL of alcohol in a suitable flask. Add 3.5 g of Raneys nickelaluminum catalyst, and connect to a suitable reflux condenser. Chill the flask in an ice bath, and add 75 mL of 2.5 N sodium hydroxide through the condenser. When the reaction has subsided, remove the ice bath. After 10 min, heat the flask gently, gradually increasing the heat until the mixture refluxes rapidly. After 90 min from the time of the addition of the alkali, discontinue heating, cool, and rinse the condenser with water, collecting the rinsings in the flask. Transfer the liquid to a 200-mL volumetric flask, wash the residue with water, and add the washing to the volumetric flask. Dilute with water to volume. Transfer 100.0 mL of this solution to a beaker, neutralize, then acidify (using congo red as the indicator) by adding nitric acid dropwise with mixing. Analysis: Titrate with 0.1 N silver nitrate VS, using silver and calomel electrodes. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Cl. Acceptance criteria: 20.6%21.2%, calculated on the dried basis IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.15% HEAVY METALS, Method II 231: NMT 0.002% Organic Impurities PROCEDURE Standard solution A: 100 g/mL of USP Chlorzoxazone Related Compound A RS (2-amino-4-chlorophenol) in methanol Standard solution B: 50 g/mL of p-chlorophenol in methanol Sample solution: 20 mg/mL in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Hexane and dioxane (63:37) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Locate the spots on the plate by examination under short-wavelength UV light.
Official Monographs / Chlorzoxazone 49

Acceptance criteria: Any spot of the Sample solution, other than one corresponding to chlorzoxazone, does not exceed, in size or intensity, the principal spot of Standard solution A, corresponding to NMT 0.5% of any individual impurity. Expose the plate to iodine vapors in a closed chamber, and locate the spots: any spot of the Sample solution, other than one corresponding to chlorzoxazone, does not exceed, in size or intensity, the principal spot of Standard solution B, corresponding to NMT 0.25% of any individual impurity. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 189194 LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Chlorzoxazone RS USP Chlorzoxazone Related Compound A RS
Chlorzoxazone Tablets
(Comment on this Monograph)id=m17092=Chlorzoxazone Tablets=Chl-Cy-Monos.pdf) DEFINITION Chlorzoxazone Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C7H4ClNO2. IDENTIFICATION A. ULTRAVIOLET ABSORPTION Sample solution: Disperse a portion of powdered Tablets, equivalent to 100 mg of chlorzoxazone, in 100 mL of methanol, shake for 15 min, and filter. Dilute in methanol to 0.02 mg/mL of chlorzoxazone. Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Chlorzoxazone RS, concomitantly measured. B. The chromatogram of the Sample solution exhibits a major peak for chlorzoxazone, the retention time of which corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Glacial acetic acid and water (1:99) Mobile phase: Acetonitrile, water, and glacial acetic acid (30:70:1) Internal standard solution: 1.25 mg/mL of phenacetin in acetonitrile Standard stock solution: 1.25 mg/mL of USP Chlorzoxazone RS in Mobile phase Standard solution: 5.0 mL of Standard stock solution and 10.0 mL of Internal standard solution in a 50-mL volumetric flask; dilute with Solution A to volume. Sample stock solution: Transfer the equivalent to 125 mg of chlorzoxazone, from powdered Tablets (NLT 20), to a 100-mL volumetric flask. Add 70 mL of acetonitrile, and shake by mechanical means for about 30 min. Dilute with acetonitrile to volume. Filter a portion of this solution, discarding the first 10 mL of the filtrate. Sample solution: Transfer 5.0 mL of the Sample stock solution to a 50-mL volumetric flask. Add 10.0 mL of the Internal standard solution and dilute with Solution A to volume. System suitability stock solution: 8.5 mg/mL of pchlorophenol in acetonitrile
50
Chlorzoxazone / Official Monographs
USP 32
System suitability solution: Transfer 1 mL of System suitability stock solution and 4 mL of Standard stock solution and 10 mL of Internal standard solution to a 50-mL volumetric flask. Dilute with Solution A to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for phenacetin, chlorzoxazone, and p-chlorophenol are about 0.7, 1.0, and 1.2, respectively.] Suitability requirements Resolution: NLT 2.0 between the chlorzoxazone peak and the p-chlorophenol peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H4ClNO2 in the Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak area ratio of chlorzoxazone to the phenacetin peak from the Sample solution = peak area ratio of chlorzoxazone to the RS phenacetin peak from the Standard solution = concentration of USP Chlorzoxazone RS in the CS Standard solution (mg/mL) CU = nominal concentration of chlorzoxazone in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 [NOTEUse 2-L vessels for this test.] Medium: pH 6.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions), 1800 mL Apparatus 2: 75 rpm Time: 60 min Analytical wavelength: UV 284 nm Standard solution: USP Chlorzoxazone RS in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution, and filter. Tolerances: NLT 75% (Q) of the labeled amount of C7H4ClNO2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Chlorzoxazone RS RU
Cholecalciferol
(Comment on this Monograph)id=m17120=Cholecalciferol=ChlCy-Monos.pdf)
C27H44O 9,10-Secocholesta-5,7,10(19)-trien-3-ol, (3,5Z,7E)-; Cholecalciferol [67-97-0]. DEFINITION Cholecalciferol contains NLT 97.0% and NMT 103.0% of C27H44O.
384.64
IDENTIFICATION A. INFRARED ABSORPTION 197K [NOTEThe range is 212 m.] B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 265 nm Medium: Alcohol Sample solution: 10 g/mL Acceptance criteria: Absorptivities do not differ by more than 3.0%. C. PROCEDURE To a solution of 0.5 mg in 5 mL of chloroform, add 0.3 mL of acetic anhydride and 0.1 mL of sulfuric acid, and shake vigorously. Acceptance criteria: A bright red color is produced, and it rapidly changes through violet and blue to green. D. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 [NOTEPrepare the Standard solution and Sample solution without heating, and handle without delay.] Diluent: 10 mg/mL squalane in chloroform Standard solution: 50 mg/mL of USP Cholecalciferol RS in Diluent Sample solution: 50 mg/mL of cholecalciferol in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Cyclohexane and diethyl ether (1:1) Spray reagent: A solution (1 in 50) of acetyl chloride in antimony trichloride TS Analysis Samples: Standard solution and Sample solution Apply the solutions on a line parallel to and about 2.5 cm from the bottom edge of a thin-layer chromatographic plate. Place the plate in a developing chamber containing and equilibrated with Developing solvent system. Develop the chromatogram until the solvent front has moved about
USP 32
15 cm above the line of application. Perform the development and subsequent operations in the dark. Remove the plate, allow the solvent to evaporate, and spray with Spray reagent. Acceptance criteria: The chromatogram from the Sample solution shows a yellowish orange area (cholecalciferol) having the same RF value as the area of the Standard solution, and may show below the cholecalciferol area a violet area attributed to 7-dehydrocholesterol. ASSAY PROCEDURE Dehydrated hexane: Prepare a chromatographic column by packing a chromatographic tube, 60- 8-cm in diameter, with 500 g of 50- to 250-m chromatographic siliceous earth, activated by drying at 150 for 4 h (see Chromatography 621, Column Adsorption Chromatography). Pass 500 mL of hexanes through the column, and collect the eluate in a glass-stoppered flask. Mobile phase: N-amyl alcohol and Dehydrated hexane (3:997) [NOTEThe ratio of components and the flow rate may be varied to meet the System suitability requirements.] [NOTEUse low-actinic glassware for all the solutions, and prepare solutions fresh daily.] Standard stock solution: 0.6 mg/mL of USP Cholecalciferol RS in toluene [NOTEDissolve without heat.] Standard solution: 120 g/mL from Standard stock solution in Mobile phase Sample stock solution: 0.6 mg/mL of Cholecalciferol in toluene [NOTEDissolve without heat.] Sample solution: 120 g/mL from Sample stock solution in Mobile phase System suitability solution: 25 mg/mL of USP Vitamin D Assay System Suitability RS in toluene and Mobile phase (1:1). Heat this solution, under reflux, at 90 for 45 min, and cool. This solution contains cholecalciferol, precholecalciferol, and trans-cholecalciferol. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L3 Injection size: 510 L System suitability Sample: System suitability solution [NOTEThe relative retention times for precholecalciferol, trans-cholecalciferol, and cholecalciferol are 0.4, 0.5, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.0 between trans-cholecalciferol and precholecalciferol Relative standard deviation: NMT 2.0% for the cholecalciferol peak from five injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C27H44O in the portion of Cholecalciferol taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cholecalciferol RS in the Standard solution (g/mL) = concentration of Cholecalciferol in the Sample CU solution (g/mL) Acceptance criteria: 97.0%103.0% rU rS CS SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +105 to +112 Sample solution: 5 mg/mL, in alcohol. Prepare the solution without delay, using Cholecalciferol from a container
Official Monographs / Cholecalciferol 51

opened NMT 30 min, and determine the optical rotation within 30 min after the solution has been prepared. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in hermetically sealed containers under nitrogen, in a cool place and protected from light. USP REFERENCE STANDARDS 11 USP Cholecalciferol RS USP Vitamin D Assay System Suitability RS
Cholecalciferol Solution
(Comment on this Monograph)id=m17125=Cholecalciferol Solution=Chl-Cy-Monos.pdf) DEFINITION Cholecalciferol Solution is a solution of Cholecalciferol in an edible vegetable oil, in Polysorbate 80, or in Propylene Glycol. It contains NLT 90.0% and NMT 120.0% of the labeled amount of C27H44O. ASSAY PROCEDURE Mobile phase: Hexane and pentanol (9.97:0.03) Standard solution A: Dissolve USP Cholecalciferol RS in toluene, and dilute with Mobile phase to 500 g/mL. [NOTEPrepare this solution fresh daily.] Standard solution B: Dilute Standard solution A with Mobile phase to 50 g/mL. [NOTEReserve a portion of this solution for the determination of the Precholecalciferol response factor.] Standard solution: Dilute Standard solution B with Mobile phase to 5 g/mL. [NOTEStore at a temperature not above 0.] Sample solution: Dilute equivalent to 250 g of cholecalciferol from Cholecalciferol Solution with Mobile phase to volume in a 50-mL volumetric flask. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L3 Flow rate: 2 mL/min Injection size: 10 L Calibration Sample: Standard solution Record the peak responses. Calculate the cholecalciferol response factor, FC: Result = CS/rS = concentration of USP Cholecalciferol RS in the Standard solution (g/mL) rS = peak response of cholecalciferol Precholecalciferol response factor Precholecalciferol solution: Transfer 5.0 mL of Standard solution B to a round-bottom flask fitted with a reflux condenser. Displace the air with nitrogen, and reflux for 1 h in a water bath under a nitrogen atmosphere to obtain a solution containing cholecalciferol and precholecalciferol. Cool, transfer the solution, with the aid of several portions of Mobile phase, to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Chromatograph the Precholecalciferol solution, and record the peak responses. Calculate the concentration, CS, in g/mL, of cholecalciferol: Result = FC rS CS
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= response factor for cholecalciferol = peak response of cholecalciferol from the Standard solution Calculate the concentration, Cpre, in g/mL, of precholecalciferol: Result = CS CS = concentration of USP Cholecalciferol RS in the Standard solution (g/mL) = concentration of cholecalciferol (g/mL) CS Calculate the response factor, Fpre, for precholecalciferol: CS Result = Cpre/rp FC rS
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Analysis: Titrate the Sample solution with 0.1 N silver nitrate VS using a silver-glass electrode system. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of Cl. Acceptance criteria: 13.0%17.0% of Cl, calculated on the dried basis. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 0.002% SPECIFIC TESTS PH 791: 4.06.0, in a slurry (1 in 100) LOSS ON DRYING 731: Dry over phosphorus pentoxide at a pressure not exceeding 50 mm of mercury at 70 for 16 h: it loses NMT 12.0% of its weight. DIALYZABLE QUATERNARY AMINES pH 9.2 buffer: 3.80 mg/mL of sodium borate decahydrate Bromothymol blue solution: 1.5 mg/mL of bromothymol blue and 4.05 mg/mL of sodium carbonate in water Standard stock solution: 0.2 0.01 mg/mL of 60% benzyltrimethylammonium chloride [NOTEPrepare this solution fresh.] Standard solution: Cut a 20- to 25-cm piece of cellulose dialysis tubing having a molecular weight cut-off that falls within the 6,00014,000 range and a dry flat width of 59 cm, and place it in water to hydrate until pliable, appropriately sealing one end. Pipet 5 mL of the Standard stock solution into the tubing, add 5 mL of water, appropriately seal the open end, place the tube in a suitable vessel containing 100 mL of water so that it is completely immersed in the water, and stir the fluid for 16 h to effect dialysis. Sample solution: Cut a 20- to 25-cm piece of cellulose dialysis tubing having a molecular weight cut-off that falls within the 6,00014,000 range and a dry flat width of 59 cm, and place it in water to hydrate until pliable, appropriately sealing one end. Weigh 2 0.01 g of Cholestyramine Resin, and carefully transfer the specimen into the tubing, taking care to ensure that none adheres to the upper walls of the tubing. Add 10 mL of water to the contents of the tube, appropriately seal the open end, and place the tube in a suitable vessel containing 100 mL of water so that it is completely immersed in the water. Stir the fluid for 16 h to effect dialysis. Analysis: Pipet the following into each of three separators: Separator 15 mL of Standard solution, Separator 25 mL of Sample solution, and Separator 35 mL of water. To each separator, add 5 mL of pH 9.2 buffer, 1 mL of Bromothymol blue solution, and 10 mL of chloroform. Shake each separator vigorously for 1 min, allow the phases to separate until the chloroform phase is clear, and collect the chloroform extracts in separate 25-mL volumetric flasks. Repeat the extraction process with a second 10-mL portion of chloroform, and combine with the previous extracts. Dilute each solution with chloroform to volume, if necessary. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 420 nm Blank: Solution from Separator 3 Analysis: Concomitantly determine the absorbances of the Sample solution and the Standard solution. Acceptance criteria: The absorbance of the Sample solution does not exceed that of the Standard solution (0.05% as benzyltrimethylammonium chloride).
A
= concentration of precholecalciferol (g/mL) Cpre = peak response of precholecalciferol rp System suitability Sample: Precholecalciferol solution [NOTEThe relative retention times for precholecalciferol and cholecalciferol are about 0.4 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.0 between the precholecalciferol peak and the choleciferol peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C27H44O in the Solution taken: Result = (FC rC) + (Fpre rpre) 100/L = response factor for cholecalciferol = peak response of cholecalciferol from the Sample solution = response factor for precholecalciferol Fpre = peak response of precholecalciferol from the rpre Sample solution L = label claim Acceptance criteria: 90.0%120.0% FC rC ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. LABELING: Label it to indicate the concentration, in mg/mL, of cholecalciferol. Label it also to state that it is to be used for manufacturing only. USP REFERENCE STANDARDS 11 USP Cholecalciferol RS
Cholestyramine Resin
(Comment on this Monograph)id=m17270=Cholestyramine Resin=Chl-Cy-Monos.pdf) Cholestyramine [11041-12-6]. DEFINITION Cholestyramine Resin is a strongly basic anion-exchange resin in the chloride form, consisting of styrene-divinylbenzene copolymer with quaternary ammonium functional groups. Each g exchanges NLT 1.8 g and NMT 2.2 g of sodium glycocholate, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K: Meets the requirements
OTHER COMPONENTS CONTENT OF CHLORIDE Sample solution: To 750 mg of Cholestyramine Resin, add 100 mL of water and 50 mg of potassium nitrate. Add, with stirring, 2 mL of nitric acid.
suitable tubing is Spectra/Por 1, Item #132 665, available from Spectrum Laboratories, Inc. (www.spectrapor.com), or equivalent.
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EXCHANGE CAPACITY Mobile phase: Acetonitrile and 0.08 M monobasic potassium phosphate (7:13). Adjust with phosphoric acid to a pH of 3.0 (see Chromatography 621, System Suitability). Potassium phosphate buffer: 4 mg/mL of monobasic potassium phosphate and 12 mg/mL of dibasic potassium phosphate Sodium glycocholate solution: 30 mg/mL of sodium glycocholate in the Potassium phosphate buffer Reference solution: Dilute Sodium glycocholate solution in water to 1.25 mg/mL. Standard solution: Transfer 100 mg of USP Cholestyramine Resin RS to a 25-mL conical flask. Pipet 15.0 mL of the Sodium glycocholate solution into the flask, and stir by mechanical means for 2 h. Transfer the contents to a centrifuge tube, and centrifuge for 15 min. Transfer 5.0 mL of the supernatant to a 50-mL volumetric flask, and dilute to volume. System suitability solution: 0.6 mg/mL of sodium glycocholate and 0.3 mg/mL of taurodeoxycholic acid in water Sample solution: Transfer 100 mg of anhydrous Cholestyramine Resin to a 25-mL conical flask. Pipet 15.0 mL of the Sodium glycocholate solution into the flask, and stir by mechanical means for 2 h. Transfer the contents to a centrifuge tube, and centrifuge for 15 min. Transfer 5.0 mL of the supernatant to a 50-mL volumetric flask, and dilute to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Samples: Reference solution and System suitability solution Suitability requirements Resolution: NLT 1.5 between sodium glycocholate and taurodeoxycholic acid, System suitability solution Tailing factor: NMT 2.5, Reference solution Relative standard deviation: NMT 1.5%, Reference solution Analysis Samples: Reference solution, Standard solution, and Sample solution Calculate the quantity, in mg, of sodium glycocholate absorbed on each g of the Resin taken: Result = M(2.5rR rU)WS/(2.5rR rS)WU = stated value of sodium glycocholate absorbed per g of USP Cholestyramine Resin RS = peak response from the Reference solution rR = peak response from the Sample solution rU = weight of USP Cholestyramine Resin RS taken to WS prepare the Standard solution (mg) = peak response from the Standard solution rS = weight of Cholestyramine Resin, calculated on WU the dried basis, taken to prepare the Sample solution (mg) Acceptance criteria: Each g exchanges NLT 1.8 g and NMT 2.2 g of sodium glycocholate, calculated on the dried basis. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cholestyramine Resin RS M
Official Monographs / Cholestyramine 53
Cholestyramine for Oral Suspension

(Comment on this Monograph)id=m17280=Cholestyramine for Oral Suspension=Chl-Cy-Monos.pdf) DEFINITION Cholestyramine for Oral Suspension is a mixture of Cholestyramine Resin with suitable excipients and coloring and flavoring agents. It contains NLT 85.0% and NMT 115.0% of the labeled amount of dried cholestyramine resin. IDENTIFICATION INFRARED ABSORPTION Sample: Transfer the equivalent to 500 mg of dried cholestyramine resin from Oral Suspension to a suitable flask, add 100 mL of 0.1 N hydrochloric acid, stir to suspend the solid, and heat on a steam bath for 10 min. Filter, wash the residue with three 50-mL portions of water, and dry at 70 and at a pressure not exceeding 50 mm of mercury for 16 h. Acceptance criteria: The IR absorption spectrum of a potassium bromide dispersion of the residue so obtained exhibits maxima only at the same wavelengths as that of a similar preparation of USP Cholestyramine Resin RS. ASSAY PROCEDURE Mobile phase: Acetonitrile and 0.08 M monobasic potassium phosphate (7:13). Adjust with phosphoric acid to a pH of 3.0. Potassium phosphate buffer: 4 mg/mL of monobasic potassium phosphate and 12 mg/mL of dibasic potassium phosphate in water Sodium glycocholate solution: 30 mg/mL of sodium glycocholate in Potassium phosphate buffer Reference solution: Dilute Sodium glycocholate solution in water to 1.25 mg/mL. Standard solution: Transfer 100 mg of USP Cholestyramine Resin RS to a 25-mL conical flask. Pipet 15.0 mL of Sodium glycocholate solution into the flask, and stir by mechanical means for 2 h. Transfer the contents to a centrifuge tube, and centrifuge for 15 min. Transfer 5.0 mL of the supernatant to a 50-mL volumetric flask, and dilute to volume. System suitability solution: 0.6 mg/mL of sodium glycocholate and 0.3 mg/mL of taurodeoxycholic acid in water Sample solution: Transfer the equivalent to 100 mg of cholestyramine resin from Oral Suspension to a 25-mL conical flask. Pipet 15.0 mL of Sodium glycocholate solution into the flask, and stir by mechanical means for 2 h. Transfer the contents to a centrifuge tube, and centrifuge for 15 min. Transfer 5.0 mL of the supernatant to a 50-mL volumetric flask, and dilute to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: Reference solution and System suitability solution Suitability requirements Resolution: NLT 1.5 between sodium glycocholate and taurodeoxycholic acid, System suitability solution
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solution for 5 min. Cool the solution slightly, and add 5 mL of nickel sulfate solution (1 in 20). Boil the solution until no more oxygen is evolved, cool, and add 2 N sulfuric acid dropwise until the color of the solution changes from yellow to orange. Add to the flask a freshly prepared solution of 4 g of potassium iodide and 2 g of sodium bicarbonate in 100 mL of water, then add 6 mL of hydrochloric acid. Immediately insert the stopper in the flask, and allow to stand in the dark for 10 min. Rinse the stopper and the sides of the flask with a few mL of water. Analysis: Titrate the liberated iodine with 0.1 N sodium thiosulfate VS to an orange color. Add 3 mL of starch TS, and continue the titration to a blue-green endpoint. Each mL of 0.1 N sodium thiosulfate is equivalent to 8.882 mg of CrCl3 6H2O. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Sulfate 221 Sample solution: 2.0 g in 10 mL of water, add 1 mL of 3 N hydrochloric acid, filter if necessary to obtain a clear solution, wash the filter with two 5-mL portions of water, and dilute with water to 40 mL Control solution: Prepare as Sample solution, but use 1.0 g of the substance under test. After the filtration step, add 0.10 mL of 0.020 N sulfuric acid. Analysis: To each solution add 3 mL of barium chloride TS, mix, and allow to stand overnight. Decant most of the supernatants, without disturbing the precipitates, but leaving twice the volume of liquid in the control solution as in the Sample solution. Dilute each solution with water to 25 mL, and sonicate for 1 min. Acceptance criteria: Any turbidity in the Sample solution does not exceed that in the control solution (0.01%). IRON 241: NMT 0.01% Sample solution: 10 mg/mL. Transfer 10 mL of this solution to a 100-mL color comparison tube, dilute with water to 45 mL, add 2 mL of hydrochloric acid, and mix. Analysis: Proceed as directed for Procedure, except to add 15 mL of butyl alcohol to the Sample solution and the Standard preparation at the same time that the Ammonium Thiocyanate solution is added. Shake for 30 s, and allow the layers to separate. Acceptance criteria: The color in the upper butyl alcohol layer from the Sample solution is not darker than that from the Standard preparation. SPECIFIC TESTS INSOLUBLE MATTER: Transfer 10 g to a 250-mL beaker, add 100 mL of water, cover the beaker, and heat to boiling. Digest the hot solution on a steam bath for 30 min, and filter through a tared filtering crucible of fine porosity. Rinse the beaker with hot water, passing the rinsings through the filter, and wash the filter with hot water until the last washing is colorless. Dry the filter at 105: the weight of the residue does not exceed 1 mg (0.01%). SUBSTANCES NOT PRECIPITATED BY AMMONIUM HYDROXIDE: Dissolve 2.0 g in 80 mL of water, heat the solution to boiling, and add a slight excess of ammonium hydroxide. Continue heating to remove the excess ammonia, cool, dilute with water to 100.0 mL, and mix. Pass through a retentive filter, and transfer 50.0 mL of the clear filtrate to an evaporating dish that previously has been ignited and tared. Add 0.5 mL of sulfuric acid to the filtrate, evaporate on a steam bath to dryness, heat gently to remove the excess acid, and ignite gently: the weight of the residue does not exceed 2.0 mg (0.20% as sulfate).
Tailing factor: NMT 2.5, Reference solution Relative standard deviation: NMT 1.5%, Reference solution Analysis Samples: Reference solution, Standard solution, and Sample solution Calculate the percent label claim of cholestyramine resin in the Cholestyramine for Oral Suspension taken: Result = [M (2.5 rr rU)WS]/[(2.5 rr rS) WU Q] 100/L = stated value of sodium glycocholate absorbed on USP Cholestyramine Resin RS (mg/g) = peak response from the Reference solution rr = peak response from the Sample solution rU = weight of USP Cholestyramine Resin RS taken to WS prepare the Standard solution (mg) = peak response from the Standard solution rS = weight of Oral Suspension taken to prepare the WU Sample solution (mg) Q = exchange capacity of the dried cholestyramine resin used in the manufacture of the Oral Suspension, as measured by the quantity of sodium glycocholate absorbed on the dried cholestyramine resin (mg/g) (see Cholestyramine Resin, Exchange Capacity) L = label claim of cholestyramine resin in Oral Suspension (mg/mg) Acceptance criteria: 85.0%115.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: for Weight Variation Meets the requirements M
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cholestyramine Resin RS
Chromic Chloride
(Comment on this Monograph)id=m17480=Chromic Chloride=Chl-Cy-Monos.pdf) CrCl3 6H2O Chromium chloride (CrCl3) hexahydrate; Chromium(3+) chloride hexahydrate [10060-12-5]. Anhydrous [10025-73-7]. 266.45
158.36
DEFINITION Chromic Chloride contains NLT 98.0% and NMT 101.0% of CrCl3 6H2O. IDENTIFICATION A. To 5 mL of a solution (1 in 250) in a test tube, add 1 mL of 5 N sodium hydroxide and 10 drops of 30% hydrogen peroxide, and heat gently for about 2 min: a yellow color develops. B. To 5 mL of a solution (1 in 250) in a test tube, add 5 drops of silver nitrate TS: a white, curdy precipitate is formed, and it is insoluble in nitric acid. ASSAY PROCEDURE Sample solution: 0.4 g of Chromic Chloride in 100 mL of water contained in a glass-stoppered, 500-mL conical flask, add 5 mL of 5 N sodium hydroxide, and mix. Pipet slowly 4 mL of 30% hydrogen peroxide into the flask, and boil the
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Official Monographs / Chromate 55

SPECIFIC TESTS PH 791: 1.52.5 BACTERIAL ENDOTOXINS TEST 85: Contains NMT 16.70 USP Endotoxin Units/g of chromium OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I or Type II glass. LABELING: Label the Injection to indicate that it is to be diluted to the appropriate strength with Sterile Water for Injection or other suitable fluid prior to administration. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Chromic Chloride Injection

(Comment on this Monograph)id=m17485=Chromic Chloride Injection=Chl-Cy-Monos.pdf) DEFINITION Chromic Chloride Injection is a sterile solution of Chromic Chloride in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of Cr. IDENTIFICATION The Assay, Sample solution exhibits an absorption maximum at about 360 nm when tested as directed for Analysis in the Assay. ASSAY PROCEDURE Sodium chloride solution: 27 mg/mL of sodium chloride Chromium stock solution: 2.829 mg/mL of potassium dichromate. This solution contains 1000 g/mL of chromium. [NOTEStore in a polyethylene bottle.] Standard solutions: Pipet 10 mL of the Chromium stock solution into a 1000-mL volumetric flask, and dilute with water to volume. Transfer 10.0 mL and 20.0 mL, respectively, of this stock solution to separate 100-mL volumetric flasks, and transfer 15.0 mL and 20.0 mL, respectively, of the stock solution to separate 50-mL volumetric flasks. Add 20 mL of Sodium chloride solution to each 100-mL volumetric flask, and 10 mL of Sodium chloride solution to each 50-mL volumetric flask, dilute the contents of each flask with water to volume, and mix. These Standard solutions contain, respectively, 1.0, 2.0, 3.0, and 4.0 g/mL of chromium. Sample solution: Transfer a volume of Injection equivalent to about 60 g of chromium to a 25-mL volumetric flask. From the labeled amount of sodium chloride, if any, in the Injection, calculate the amount, in mg, of sodium chloride in the volume of Injection taken, and add sufficient Sodium chloride solution to bring the total sodium chloride content of the flask to 135 mg. Dilute with water to volume, and mix. Blank: 1:5 dilution of the Sodium chloride solution Spectroscopic conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Lamp: Chromium hollow-cathode Flame: Airacetylene Absorption wavelength: Chromium emission line at 357.9 nm Analysis: Determine the absorbances of the Standard solutions and the Sample solution. Plot the absorbances of the Standard solutions versus concentration, in g/mL, of chromium, and draw the straight line best fitting the four plotted points. From the graph so obtained, determine the concentration, in g/mL, of chromium in the Sample solution. Calculate the quantity, in g, of chromium in each mL of the Injection taken: Result = 25 C/V C V = concentration of chromium in the Sample solution (g/mL) = volume of Injection taken (mL)
Sodium Chromate Cr 51 Injection

(Comment on this Monograph)id=m17530=Sodium Chromate Cr 51 Injection=Chl-Cy-Monos.pdf) Chromic acid (H251CrO4), disodium salt; Disodium chromate (Na251CrO4) [7775-11-3]. DEFINITION Sodium Chromate Cr 51 Injection is a sterile solution of radioactive chromium (51Cr) processed in the form of sodium chromate in Water for Injection. For those uses where an isotonic solution is required, Sodium Chloride may be added in appropriate amounts as provided under Injections 1. Chromium 51 is produced by the neutron bombardment of enriched chromium 50. Sodium Chromate Cr 51 Injection contains NLT 90.0% and NMT 110.0% of the labeled amount of 51Cr as sodium chromate expressed in megabecquerels (millicuries)/mL at the time indicated in the labeling. The sodium chromate content is NLT 90.0% and NMT 110.0% of the labeled amount. The specific activity is NLT 370 megabecquerels (10 millicuries)/mg of sodium chromate at the end of the expiry period. Other chemical forms of radioactivity do not exceed 10.0% of the total radioactivity. IDENTIFICATION RADIOACTIVITY, Radionuclide Identification 821: Its gammaray spectrum is identical to that of a specimen of 51Cr of known purity that exhibits a photopeak having an energy of 0.320 MeV. ASSAY PROCEDURE 1: RADIOACTIVITY 821: Using a suitable counting assembly (see Radioactivity 821, Selection of a Counting Assembly), determine the radioactivity, in MBq (or Ci)/mL, of Injection by use of a calibrated system. Acceptance criteria: 90.0%110.0% of the labeled amount of 51Cr PROCEDURE 2: SODIUM CHROMATE Standard stock solution: 3.735 mg/mL of potassium chromate in water to obtain 1.0 mg/mL of chromium solution Standard solutions: Pipet 0.25, 0.50, 0.75, 0.100, 0.125, and 0.150 mL of the Standard stock solution into separate 100-mL volumetric flasks. To each flask add 0.42 mL of 0.1 N sodium bicarbonate, and dilute with water to volume to obtain solutions having final concentrations of 0.25, 0.50, 0.75, 1.00, 1.25, and 1.50 g/mL of chromium
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as megabecquerels (millicuries)/mL at the time of calibration; a statement to indicate whether the contents are intended for diagnostic or therapeutic use; the expiration date; and the statement [CAUTIONRadioactive Material]. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay and the quantity of chromium, and also indicates that the radioactive half-life of 51Cr is 27.8 days. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Sample solution: Use the Injection. Spectrometric conditions (See Spectrophotometry and Light-Scattering 851.) Mode: Atomic absorption spectrophotometer Lamp: Chromium hollow-cathode Flame: Airacetylene (fuel-rich) Analytical wavelength: Chromium emission line at 357.7 nm Blank: 0.42 mL of 0.1 N sodium bicarbonate in a 100-mL volumetric flask, and dilute with water to volume Analysis Samples: Standard solutions, Sample solution, and Blank Use water to set the instrument to zero. Determine the absorbances of the solutions. Plot the absorbances of the Standard solutions and the Blank versus concentration, in g/mL, of chromium, and perform a regression analysis. A suitable standard curve will have an intercept between 0.002 and +0.002, and a regression coefficient of NLT 0.99. Using the standard curve so obtained, determine the concentration, C, in g/mL, of chromium in the Injection taken. Calculate the quantity of sodium chromate, in g/mL: Result = 3.115 C 3.115 = conversion factor Acceptance criteria: 90.0%110.0% of the labeled amount of sodium chromate IMPURITIES Inorganic Impurities RADIOCHEMICAL PURITY Sample: Injection Chromatographic system (See Chromatography 621, System Suitability.) Mode: Paper chromatography Absorbent: 25- 300-mm strip of chromatographic paper Application volume: Appropriately diluted Sample, so that it provides a count rate of 20,000 counts/min Developing solvent system: Dilute alcohol (95 in 100), ammonium hydroxide, and water (2:1:5) Analysis: Place the application volume about 25 mm from one end of the strip and immediately develop. Air-dry the chromatogram, and determine the radioactivity distribution by scanning the chromatogram with a suitable collimated radiation detector. Acceptance criteria: The radioactivity of the chromate band is NLT 90.0% of the total radioactivity. The RF value for the chromate band falls within 10% of the value found for a known sodium chromate specimen when determined under identical conditions. SPECIFIC TESTS PH 791: 7.58.5 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 175/V USP Endotoxin Unit/mL of the Injection, when compared with the USP Endotoxin RS, in which V is the maximum recommended total dose, in mL, at the expiration date or time. OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that it is not subject to the recommendation on Volume in Container. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or in multiple-dose containers. LABELING: Label it to include the following, in addition to the information specified for Labeling under Injections 1: the time and date of calibration; the amount of sodium chromate expressed in g/mL; the amount of 51Cr as sodium chromate expressed as total megabecquerels (millicuries) and
Chromium Cr 51 Edetate Injection

(Comment on this Monograph)id=m17531=Chromium Cr 51 Edetate Injection=Chl-Cy-Monos.pdf) Glycine, N,N-1,2-ethanediylbis[N-(carboxymethyl)]-, chromium-51 complex; (Ethylenedinitrilo)tetraacetic acid, chromium-51 complex [27849-89-4]. DEFINITION Chromium Cr 51 Edetate Injection is a sterile solution containing radioactive chromium (51Cr) in the form of a complex of chromium (III) with edetic acid, present in excess. It is made isotonic by the addition of Sodium Chloride. It contains NLT 90.0% and NMT 110.0% of the labeled amount of 51Cr as edetate complex expressed in megabecquerels (or microcuries or millicuries)/mL at the time indicated in the labeling. Other chemical forms of radioactivity do not exceed 5.0% of the total radioactivity. It may contain a suitable preservative. It contains NMT 1 mg/mL of chromium (Cr). IDENTIFICATION A. RADIOACTIVITY, Radionuclide Identification 821: Its gamma-ray spectrum is identical to that of a sample of 51Cr of known purity that exhibits a photopeak having an energy of 0.320 MeV. ASSAY RADIOACTIVITY 821 Analysis: Using a suitable counting assembly (see Radioactivity 821, Selection of a Counting Assembly), determine the radioactivity, in MBq (or Ci)/mL, of Injection by use of a calibrated system. IMPURITIES Inorganic Impurities LIMIT OF FREE CHROMIUM Standard solution: Dissolve 0.96 g of chromium potassium sulfate dodecahydrate and 2.87 g of edetate disodium in 50 mL of water, boil for 10 min, cool, adjust with 0.2 M sodium hydroxide to a pH between 3.5 and 6.5, and dilute with water to 100.0 mL to obtain 1 mg/mL of chromium solution. Sample solution: Use the Injection. Spectrometric conditions Analytical wavelength: 560 nm Blank: Water Analysis Samples: Standard solution and Sample solution Acceptance criteria: The absorbance of the Sample solution is NMT that of the Standard solution. RADIOCHEMICAL PURITY Electrolyte solution: 0.2 mg/mL of barbital sodium and 10 mg/mL of sodium nitrate in water Analysis (see Electrophoresis 726): Soak a 2.5-cm 17.0-cm 0.22-mm cellulose strip in 100 mL of Electrolyte solution for 1060 min. Remove the strip with forceps, and blot to remove excess solution. Attach the strip to the support bridge of an electrophoresis chamber containing
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Electrolyte solution. Apply to the strip about 10 L of Injection as a 3-mm band at a position 10 cm from the cathode. Attach the chamber cover, and perform the electrophoresis at 30 V/cm, using a stabilized current. Remove the strip from the chamber, and blot the ends. Using a suitable scanner and counting assembly, determine the radioactivity distribution: chromium 51Cr edetate moves about 5 cm towards the anode; and 51Cr chromic ion moves about 7 cm towards the cathode. Acceptance criteria: The radioactivity of the chromium 51Cr edetate band is NLT 95% of the total radioactivity. RADIONUCLIDIC PURITY Analysis: Using a suitable gamma-ray spectrometer (see Selection of a Counting Assembly in the Assay section under Radioactivity 821), determine the radioactivity of each radionuclidic impurity observed in the gamma-ray spectrum. Acceptance criteria Any individual impurity: NMT 0.1% Total impurities: NMT 0.3% CHEMICAL PURITY Analysis: Using a validated limit test and a known analytical technique, demonstrate the absence of any ingredients and reagents employed in the synthetic process. SPECIFIC TESTS PH 791: 3.56.5 BACTERIAL ENDOTOXINS TEST 85: NMT 175/V USP Endotoxin Unit/mL of the Injection, in which V is the maximum recommended total dose, in mL, at the expiration date or time OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that it is not subject to the recommendation on Volume in Container. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, at a temperature between 2 and 8. LABELING: Label it to include the following, in addition to the information specified for Injections 1, Labeling: the time and date of calibration; the amount of 51Cr as edetate complex expressed as total MBq (or mCi) and as MBq (or mCi)/mL at the time of calibration; the expiration date; and the statement [CAUTIONRadioactive Material]. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay and the quantity of chromium, and also indicates that the radioactive half-life of 51Cr is 27.8 days. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Official Monographs / Chymotrypsin 57

Solution A: Monobasic potassium phosphate solution and Dibasic sodium phosphate solution (38.9:61.1) [NOTEIf necessary, adjust to a pH of 7.0 by the dropwise addition of Dibasic sodium phosphate solution.] Substrate solution: Dissolve 23.7 mg of N-acetyl-L-tyrosine ethyl ester, suitable for use in assaying Chymotrypsin, in 50 mL of Solution A, with warming. When the solution is cool, dilute with additional Solution A to 100 mL. [NOTESubstrate solution may be stored in the frozen state and used after thawing, but it is important to freeze it immediately after preparation.] Sample solution: Dissolve a quantity of Chymotrypsin in 0.0012 N hydrochloric acid to yield a solution containing between 12 and 16 USP Chymotrypsin Units/mL. The dilution is correct if, during the conduct of the Assay, there is a change in absorbance of between 0.008 and 0.012 in each 30-s interval. Analysis [NOTEDetermine the suitability of the substrate and check the adjustment of the spectrophotometer by performing this Analysis using USP Chymotrypsin RS in place of the Sample solution.] Conduct the Assay in a suitable spectrophotometer equipped to maintain a temperature of 25 0.1 in the cell compartment. Determine the temperature in the reaction cell before and after the measurement of absorbance to ensure that the temperature does not change by more than 0.5. Pipet 0.2 mL of 0.0012 N hydrochloric acid and 3.0 mL of Substrate solution into a 1-cm cell. Place this cell in the spectrophotometer, and adjust the instrument so that the absorbance will read 0.200 at 237 nm. Pipet 0.2 mL of Sample solution into another 1-cm cell, add 3 mL of Substrate solution, and place the cell in the spectrophotometer. [NOTECarefully follow this order of addition, and begin timing the reaction from the addition of the Substrate solution.] Read the absorbance at 30-s intervals for NLT 5 min. Repeat the procedure on the same dilution at least once. Absolute absorbance values are less important than a constant rate of absorbance change. If the rate of change fails to remain constant for NLT 3 min, repeat the test and, if necessary, use a lower concentration. The duplicate determination at the same dilution matches the first determination in rate of absorbance change. Determine the average absorbance change per min, using only the values within the 3-min portion of the curve where the rate of absorbance change is constant. Plot a curve of absorbance against time. One USP Chymotrypsin Unit is the activity causing a change in absorbance of 0.0075/min under the conditions specified in this Assay. Calculate the number of USP Chymotrypsin Units/mg taken: Result = (A2 A1)/(0.0075 T W) = absorbance straight-line initial reading = absorbance straight-line final reading = elapsed time between the initial and final reading (min) W = weight of Chymotrypsin in the volume of solution used in determining the absorbance (mg) Acceptance criteria: NLT 1000 USP Chymotrypsin Units/mg; NLT 90.0% and NMT 110.0% of the labeled potency IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 2.5% Organic Impurities PROCEDURE: LIMIT OF TRYPSIN Chymotrypsin solution: 10 mg/mL in water Solution A: Dissolve 294 mg of calcium chloride in 40 mL of 0.20 M tris(hydroxymethyl)aminomethane, adjust with 1 A2 A1 T
Chymotrypsin
(Comment on this Monograph)id=m17600=Chymotrypsin=ChlCy-Monos.pdf) Chymotrypsin [9004-07-3]. DEFINITION Chymotrypsin is a proteolytic enzyme crystallized from an extract of the pancreas gland of the ox, Bos taurus Linn (Fam. e Bovidae). It contains NLT 1000 USP Chymotrypsin Units in each mg, calculated on the dried basis, and NLT 90.0% and NMT 110.0% of the labeled potency, as determined by the Assay. ASSAY PROCEDURE Monobasic potassium phosphate solution: 9.08 mg/mL of monobasic potassium phosphate in water Dibasic sodium phosphate solution: 9.46 mg/mL of anhydrous dibasic sodium phosphate in water
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Chymotrypsin / Official Monographs

N hydrochloric acid to a pH of 8.1, and dilute with water to 100 mL. Substrate solution: Transfer 98.5 mg of p-toluenesulfonylL-arginine methyl ester hydrochloride, suitable for use in assaying trypsin, to a 25-mL volumetric flask. Add 5 mL of Solution A, and swirl until the substrate dissolves. Add 0.25 mL of methyl red-methylene blue TS, and dilute with water to volume. Analysis [NOTEDetermine the suitability of the substrate by performing this analysis using the appropriate amount of USP Trypsin Crystallized RS in place of the solution under test.] By means of a micropipet, transfer 50 L of Chymotrypsin solution to a depression on a white spot plate. Add 0.2 mL of Substrate solution. Acceptance criteria: No purple color develops within 3 min (NMT 1% of trypsin).
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ASSAY PROCEDURE Monobasic potassium phosphate solution: 9.08 mg/mL of monobasic potassium phosphate in water Dibasic sodium phosphate solution: 9.46 mg/mL of anhydrous dibasic sodium phosphate in water Solution A: Monobasic potassium phosphate solution and Dibasic sodium phosphate solution (38.9:61.1). If necessary, adjust to a pH of 7.0 by the dropwise addition of Dibasic sodium phosphate solution. Substrate solution: Dissolve 23.7 mg of N-acetyl-L-tyrosine ethyl ester, suitable for use in assaying chymotrypsin, in 50 mL of Solution A, with warming. When the solution is cool, dilute with additional Solution A to 100 mL. [NOTESubstrate solution may be stored in the frozen state and used after thawing, but it is important to freeze it immediately after preparation.] Sample stock solution: Dissolve the contents of 1 vial of Ophthalmic Solution in 5.0 mL of 0.0012 N hydrochloric acid. Sample solution: Dilute a volume (V2, in mL) of the Sample stock solution, equivalent to 300 USP Chymotrypsin Units, with 0.0012 N hydrochloric acid to 25.0 mL. Analysis [NOTEDetermine the suitability of the substrate and check the adjustment of the spectrophotometer by performing this Analysis using USP Chymotrypsin RS in place of the Sample solution.] Conduct the Assay in a suitable spectrophotometer equipped to maintain a temperature of 25 0.1 in the cell compartment. Determine the temperature in the reaction cell before and after the measurement of absorbance to ensure that the temperature does not change by more than 0.5. Pipet 0.2 mL of 0.0012 N hydrochloric acid and 3.0 mL of Substrate solution into a 1-cm cell. Place this cell in the spectrophotometer, and adjust the instrument so that the absorbance will read 0.200 at 237 nm. Pipet 0.2 mL of Sample solution into another 1-cm cell, add 3 mL of Substrate solution, and place the cell in the spectrophotometer. [NOTECarefully follow this order of addition, and begin timing the reaction from the addition of the Substrate solution. Read the absorbance at 30-s intervals for NLT 5 min. Repeat the procedure on the same dilution at least once. Absolute absorbance values are less important than a constant rate of absorbance change. If the rate of change fails to remain constant for NLT 3 min, repeat the test and, if necessary, use a lower concentration. The duplicate determination at the same dilution matches the first determination in rate of absorbance change. Determine the average absorbance change per min, using only the values within the 3-min portion of the curve where the rate of absorbance change is constant. Plot a curve of absorbance against time. One USP Chymotrypsin Unit is the activity causing a change in absorbance of 0.0075/min under the conditions specified in this Assay. Calculate the percentage of label claim of USP Chymotrypsin Units in a vial: ] Result = U (V1/V2) (A2 A1)/(T 2.4 0.0075/min) (100/L) U V1 V2 A2 A1 T = total USP Chymotrypsin Units in the Sample solution, 300 = volume of the Sample stock solution (mL), 5 = volume (mL) as defined under Sample solution = absorbance straight-line initial reading = absorbance straight-line final reading = elapsed time (min) between the initial and final readings
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Pseudomonas aeruginosa and Salmonella species and Staphylococcus aureus. LOSS ON DRYING 731: Dry in a vacuum oven at 60 for 4 h: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Chymotrypsin RS USP Trypsin Crystallized RS
Chymotrypsin for Ophthalmic Solution

(Comment on this Monograph)id=m17610=Chymotrypsin for Ophthalmic Solution=Chl-Cy-Monos.pdf) DEFINITION Chymotrypsin for Ophthalmic Solution is sterile Chymotrypsin. When constituted as directed in the labeling, it yields a solution containing NLT 80.0% and NMT 120.0% of the labeled potency. IDENTIFICATION PROCEDURE Monobasic potassium phosphate solution: 9.08 mg/mL of monobasic potassium phosphate in water Dibasic sodium phosphate solution: 9.46 mg/mL of anhydrous dibasic sodium phosphate in water Solution A: Monobasic potassium phosphate solution and Dibasic sodium phosphate solution (38.9:61.1). If necessary, adjust to a pH of 7.0 by the dropwise addition of Dibasic sodium phosphate solution. Substrate solution: Transfer 237.0 mg of N-acetyl-L-tyrosine ethyl ester, suitable for use in assaying chymotrypsin, to a 100-mL volumetric flask, add 2 mL of alcohol, and swirl until solution is effected. Add 20 mL of Solution A, add 1 mL of methyl redmethylene blue TS, and dilute to volume. If necessary, adjust to a pH of 7.0 by the dropwise addition of Monobasic potassium phosphate solution. Sample solution: Dissolve the contents of 1 vial of Ophthalmic Solution in 1 mL of saline TS. Analysis: Transfer 0.2 mL of Sample solution to a suitable dish, and add 0.2 mL of Substrate solution. Acceptance criteria: A purple color is produced within 3 min. [NOTEThis is distinct from trypsin, which produces no purple color within 3 min.]
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= number of USP Chymotrypsin Units in the solution on which the absorbance was determined L = label potency of entire vial (USP Chymotrypsin Units) Acceptance criteria: 80.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements Analysis: Assay 10 individual units as directed in the Assay, and calculate the average of the 10 results. Acceptance criteria: The average is NLT 80.0% and NMT 120.0% of the labeled amount. The contents of NMT 2 vials deviate by more than 10% from the average content. The contents of none of the vials deviate by more than 15% from the average. IMPURITIES LIMIT OF TRYPSIN Chymotrypsin solution: 10 mg/mL in water from Ophthalmic Solution Solution A: Dissolve 294 mg of calcium chloride in 40 mL of 0.20 M tris(hydroxymethyl)aminomethane, adjust with 1 N hydrochloric acid to a pH of 8.1, and dilute to 100 mL. Substrate solution: Transfer 98.5 mg of p-toluenesulfonyl-Larginine methyl ester hydrochloride, suitable for use in assaying trypsin, to a 25-mL volumetric flask. Add 5 mL of Solution A, and swirl until the substrate dissolves. Add 0.25 mL of methyl redmethylene blue TS, and dilute to volume. Analysis [NOTEDetermine the suitability of the substrate by performing the Analysis using the appropriate amount of USP Trypsin Crystallized RS in place of the Sample solution.] By means of a micropipet, transfer 50 L of Chymotrypsin solution to a depression on a white spot plate. Add 0.2 mL of the Substrate solution. Acceptance criteria: No purple color develops within 3 min (NMT 1% of trypsin). SPECIFIC TESTS PH 791: 4.38.7, in the solution constituted as directed in the labeling STERILITY TESTS 71: It meets the requirements. COMPLETENESS OF SOLUTION 641: It dissolves in the solvent and in the concentration recommended in the labeling to yield a clear solution. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Chymotrypsin RS USP Trypsin Crystallized RS 2.4
Official Monographs / Ciclopirox 59
Ciclopirox
(Comment on this Monograph)id=m17620=Ciclopirox=Chl-CyMonos.pdf)
C12H17NO2 2(1H)-Pyridinone, 6-cyclohexyl-1-hydroxy-4-methyl-; 6-Cyclohexyl-1-hydroxy-4-methyl-2(1H)-pyridone [29342-05-0]. DEFINITION Ciclopirox contains NLT 98.0% and NMT 101.0% of C12H17NO2, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K
207.27
ASSAY PROCEDURE Sample: 150 mg Analysis: Dissolve the Sample in 20 mL of methanol. Add 20 mL of water. Titrate with 0.1 N sodium hydroxide VS. Carry out a blank test. Determine the factor of the 0.1 N sodium hydroxide VS using 100 mg of benzoic acid and titrate under the conditions prescribed above. Each mL of 0.1 N sodium hydroxide is equivalent to 20.73 mg of C12H17NO2. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE [NOTECarry out the operations avoiding exposure to actinic light. All materials that are in direct contact with ciclopirox (e.g., column materials, reagents, solvents, etc.) should contain only very low amounts of extractable metal cations.] Mobile phase: Acetonitrile, edetate disodium solution (0.96 in 1000), and glacial acetic acid (230:770:0.1) Diluent: Acetonitrile and Mobile phase (1:9) Rinsing solution: Acetonitrile, acetylacetone, glacial acetic acid, and water (500:1:1:500) Standard stock solution: 1.5 mg/mL each of USP Ciclopirox Related Compound A RS and USP Ciclopirox Related Compound B RS in Diluent
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Ciclopirox / Official Monographs

Standard solution A: Dilute 1.0 mL of Standard stock solution with Diluent to 200 mL. Standard solution B: Dilute 2.0 mL of Standard solution A with Diluent to 10 mL. Sample solution: 30 mg of Ciclopirox in a mixture of 2 mL of acetonitrile and 15 mL of Mobile phase. If necessary, use an ultrasonic bath to dissolve. Dilute with Mobile phase to 20.0 mL. System suitability solution: Standard stock solution and Sample solution (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm and 298 nm [NOTECiclopirox related compound A intense absorbs strongly at 220 nm. Ciclopirox, ciclopirox related compound B, and 6-cyclohexyl-4-methyl-2(1H)-pyridone absorb strongly at 298 nm.] Column: 4.0-mm 8-cm; packing L10 Flow rate: 0.7 mL/min Injection size: 10 L System suitability Samples: Standard solution B, Sample solution, and System suitability solution at 298 nm [NOTEThe relative retention times for ciclopirox related compound A, 6-cyclohexyl-4-methyl-2(1H)-pyridone, ciclopirox, and ciclopirox related compound B are 0.5, 0.9, 1.0, and 1.3, respectively.] Suitability requirements Resolution: NLT 2.0 between the ciclopirox related compound B peak and ciclopirox peak, System suitability solution Signal-to-noise ratio: NLT 3 for ciclopirox related compound B, Standard solution B Tailing factor: NMT 2.0 factor for the ciclopirox peak, Sample solution Analysis Samples: Standard solution A, Standard solution B, and Sample solution [NOTEIn order to ensure desorption of disruptive metal ions, every new column must be rinsed with the Rinsing solution over a period of NLT 15 h and then with the Mobile phase for NLT 5 h with a flow rate of 0.2 mL/min. The chromatographic run time is NLT 2.5 times the retention time of the ciclopirox peak.] Acceptance criteria: The peak response at 220 nm of the ciclopirox related compound A peak in the chromatogram of the Sample solution is NMT the peak response at 220 nm of the corresponding peak in the chromatogram of Standard solution A (0.5% with reference to ciclopirox). The sum of responses at 298 nm of the peaks in the chromatogram of the Sample solution is NMT the peak response at 298 nm of the ciclopirox related compound B peak in the chromatogram of Standard solution A (0.5% with reference to ciclopirox). [NOTEAt 298 nm disregard any peak due to the solvent and any peak with a response less than the response of the ciclopirox related compound B peak in the chromatogram of Standard solution B (0.1% with reference to ciclopirox).] USP Ciclopirox Related Compound B RS
USP 32
Ciclopirox Olamine
(Comment on this Monograph)id=m17630=Ciclopirox Olamine=Chl-Cy-Monos.pdf)
268.35 C12H17NO2 C2H7NO 2(1H)-Pyridinone, 6-cyclohexyl-1-hydroxy-4-methyl-, compound with 2-aminoethanol (1:1); 6-Cyclohexyl-1-hydroxy-4-methyl-2(1H)-pyridone compound with 2-aminoethanol (1:1) [41621-49-2]. DEFINITION Ciclopirox Olamine contains NLT 97.5% and NMT 101.5% of ciclopirox olamine (C12H17NO2 C2H7NO). IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 200 mg Analysis: Dissolve the Sample in 2 mL of methanol and add 38 mL of water. Titrate with 0.1 N sodium hydroxide VS. Perform a blank determination, and make any necessary corrections. Determine the factor of the 0.1 N sodium hydroxide VS using 100 mg of benzoic acid and titrate under the conditions prescribed above. Each mL of 0.1 N sodium hydroxide is equivalent to 26.84 mg of C12H17NO2 C2H7NO. Acceptance criteria: 97.5%101.5% OTHER COMPONENTS MONOETHANOLAMINE CONTENT Analysis: Dissolve 300 mg in 25 mL of glacial acetic acid. Titrate with 0.1 N perchloric acid VS. Perform a blank determination and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 6.108 mg of C2H7NO. Acceptance criteria: 223230 mg/g of C12H17NO2 C2H7NO found in the Assay IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE [NOTECarry out the operations avoiding exposure to actinic light. All materials that are in direct contact with Ciclopirox Olamine (e.g., column materials, reagents, solvents, etc.) should contain only very low amounts of extractable metal cations.] Mobile phase: Acetonitrile, edetate disodium solution (0.96 in 1000), and glacial acetic acid (230:770:0.1) Diluent: Acetonitrile and Mobile phase (1:9) Rinsing solution: Acetonitrile, acetylacetone, glacial acetic acid, and water (500:1:1:500) Standard stock solution: 1.5 mg/mL each of USP Ciclopirox Related Compound A RS and USP Ciclopirox Related Compound B RS in Diluent Standard solution A: Dilute 1.0 mL of Standard stock solution with Diluent to 200 mL.
SPECIFIC TESTS LOSS ON DRYING 731: Dry it in a vacuum to constant weight: it loses NMT 1.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, protected from light. Store at a temperature between 15 and 30. USP REFERENCE STANDARDS 11 USP Ciclopirox RS USP Ciclopirox Related Compound A RS
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Standard solution B: Dilute 2.0 mL of Standard solution A with Diluent to 10 mL. Sample solution: 40 mg of Ciclopirox Olamine in a mixture of 2 mL of acetonitrile, 20 L of glacial acetic acid, and 15 mL of Mobile phase. If necessary, use an ultrasonic bath to dissolve. Dilute with Mobile phase to 20.0 mL. System suitability solution: Standard stock solution and Sample solution (1:1) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm and 298 nm [NOTECiclopirox related compound A intense absorbs strongly at 220 nm. Ciclopirox, Ciclopirox related compound B, and 6-cyclohexyl-4-methyl-2(1H)-pyridone absorb strongly at 298 nm.] Column: 4.0-mm 8-cm; packing L10 Flow rate: 0.7 mL/min Injection size: 10 L System suitability Samples: Standard solution B, Sample solution, and System suitability solution at 298 nm [NOTEThe relative retention times for ciclopirox related compound A, 6-cyclohexyl-4-methyl-2(1H)-pyridone, ciclopirox, and ciclopirox related compound B are 0.5, 0.9, 1.0, and 1.3, respectively.] Suitability requirements Resolution: NLT 2.0 between the ciclopirox related compound B peak and ciclopirox peak, System suitability solution Signal-to-noise ratio: NLT 3 for ciclopirox related compound B, Standard solution B Tailing factor: NMT 2.0 factor for the ciclopirox peak, Sample solution Analysis Samples: Standard solution A, Standard solution B, and Sample solution [NOTEIn order to ensure desorption of disruptive metal ions, every new column must be rinsed with the Rinsing solution over a period of NLT 15 h and then with the Mobile phase for NLT 5 h with a flow rate of 0.2 mL/min. The chromatographic run time is NLT 2.5 times the retention time of the ciclopirox peak.] Acceptance criteria: The peak response at 220 nm of the ciclopirox related compound A peak in the chromatogram of the Sample solution is NMT the peak response at 220 nm of the corresponding peak in the chromatogram of Standard solution A (0.5% with reference to ciclopirox). The sum of responses at 298 nm of the peaks in the chromatogram of the Sample solution is NMT the peak response at 298 nm of the ciclopirox related compound B peak in the chromatogram of Standard solution A (0.5% with reference to ciclopirox). [NOTEAt 298 nm disregard any peak due to the solvent and any peak with a response less than the response of the ciclopirox related compound B peak in the chromatogram of Standard solution B (0.1% with reference to ciclopirox).] SPECIFIC TESTS PH 791: 8.09.0, in a mixture with water (1:100) ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. Store between 5 and 25. USP REFERENCE STANDARDS 11 USP Ciclopirox Olamine RS USP Ciclopirox Related Compound A RS USP Ciclopirox Related Compound B RS
Official Monographs / Ciclopirox 61
Ciclopirox Olamine Cream

(Comment on this Monograph)id=m17633=Ciclopirox Olamine Cream=Chl-Cy-Monos.pdf) DEFINITION Ciclopirox Olamine Cream contains NLT 90.0% and NMT 110.0% of the labeled amount of ciclopirox olamine (C12H17NO2 C2H7NO). IDENTIFICATION ULTRAVIOLET ABSORPTION Sample solution: Dilute 4 mL of the Sample solution obtained as directed in the Assay with a mixture of methanol and 6.25 N sodium hydroxide (123:2) to make 100 mL. Acceptance criteria: The UV absorption spectrum of the solution so obtained exhibits maxima and minima at the same wavelengths as that of a similar solution prepared from the Standard solution obtained as directed in the Assay, concomitantly measured. ASSAY PROCEDURE Ferrous sulfate solution: 24 mg/mL of ferrous sulfate in glacial acetic acid and water (0.6:24.4) Standard solution: 0.2 mg/mL of USP Ciclopirox Olamine RS in methanol Sample solution: Equivalent to 0.2 mg/mL of ciclopirox olamine from Cream, in methanol [NOTEShake by mechanical means for about 10 min, centrifuge, and use the supernatant.] Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 440 nm Cell: 1 cm Analysis: Transfer 4.0 mL of the Standard solution, 4.0 mL of the Sample solution, and 4.0 mL of methanol to provide a blank, to separate 25-mL volumetric flasks. Add 15 mL of methanol to each flask. Then to each flask add 1.0 mL of Ferrous sulfate solution, and dilute with methanol to volume. Store the flasks in the dark for 1 h. Determine the absorbances of the resulting solutions at the wavelength of maximum absorbance, 440 nm. Calculate the percentage of C12H17NO2 C2H7NO in the Cream taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% AU AS CS CU OTHER COMPONENTS CONTENT OF BENZYL ALCOHOL (IF PRESENT) Diluent: Chloroform and methanol (4:1) Internal standard solution: 1.75 mg/mL of 1-nonyl alcohol in Diluent Standard solution: 2 mg/mL of USP Benzyl Alcohol RS in Diluent. Transfer 5.0 mL to a 50-mL volumetric flask, add 5.0 mL of Internal standard solution, and dilute with Diluent to volume. Sample solution: Transfer 1.0 g of Cream to a 50-mL volumetric flask, and add 30 mL of Diluent. Add 5.0 mL of Internal standard solution, and dilute with Diluent to volume. = = = =
62
Ciclopirox / Official Monographs
USP 32
IDENTIFICATION A. ULTRAVIOLET ABSORPTION Sample solution: Dilute 4 mL of the Sample solution obtained as directed in the Assay with a mixture of methanol and 6.25 N sodium hydroxide (123:2) to make 100 mL. Acceptance criteria: The UV absorption spectrum of the solution so obtained exhibits maxima and minima at the same wavelengths as that of a similar solution prepared from the Standard solution obtained as directed in the Assay, concomitantly measured. ASSAY PROCEDURE Ferrous sulfate solution: 24 mg/mL of ferrous sulfate in glacial acetic acid and water (0.6:24.4) Standard solution: 0.2 mg/mL of USP Ciclopirox Olamine RS in methanol Sample solution: Equivalent to 0.2 mg/mL of ciclopirox olamine from Topical Suspension, in methanol [NOTEShake by mechanical means for about 10 min, centrifuge, and use the supernatant.] Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 440 nm Cell: 1 cm Analysis: Transfer 4.0 mL of the Standard solution, 4.0 mL of the Sample solution, and 4.0 mL of methanol to provide a blank, to separate 25-mL volumetric flasks. Add 15 mL of methanol to each flask. Then to each flask add 1.0 mL of Ferrous sulfate solution, mix, and dilute with methanol to volume. Store the flasks in the dark for 1 h. Determine the absorbances of the resulting solutions at the wavelength of maximum absorbance, 440 nm. Calculate the percentage of C12H17NO2 C2H7NO in the portion of Topical Suspension taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Ciclopirox Olamine RS in the Standard solution (mg/mL) = nominal concentration of ciclopirox olamine in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% AU AS CS OTHER COMPONENTS CONTENT OF BENZYL ALCOHOL Diluent: Chloroform and methanol (4:1) Internal standard solution: 1.75 mg/mL of 1-nonyl alcohol in Diluent Standard solution: 2 mg/mL of USP Benzyl Alcohol RS in Diluent. Transfer 5.0 mL to a 50-mL volumetric flask, add 5.0 mL of Internal standard solution, and dilute with Diluent to volume. Sample solution: Transfer 1.0 g of Topical Suspension to a 50-mL volumetric flask, add 30 mL of Diluent, and mix. Add 5.0 mL of Internal standard solution, and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 2-m glass column packed with 3% phase G3 on 100- to 120-mesh support S1AB
Chromatographic system Mode: GC Detector: Flame ionization Column: 4-mm 2-m glass column packed with 3% phase G3 on 100- to 120-mesh support S1AB Temperature Column: 100 Injector: 315 Detector: 315 Carrier gas: Nitrogen Flow rate: 45 mL/min Injection size: 2 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.6 between the peaks Tailing factor: NMT 3.5 for the benzyl alcohol peak and the internal standard peak Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution [NOTEAfter six injections, raise the column temperature to about 300 for about 5 min, then cool to 100.] Calculate the percentage of benzyl alcohol in the Cream taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the benzyl alcohol peak to the internal standard peak from the Sample solution = peak response ratio of the benzyl alcohol peak RS to the internal standard peak from the Standard solution = concentration of benzyl alcohol in the Standard CS solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amount of benzyl alcohol PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS PH 791: 5.08.0 Analysis: Add 15 mL of boiling water, previously adjusted with 0.1 N hydrochloric acid or 0.1 N sodium hydroxide to a pH of 67, to 3.5 g of Cream in a 50-mL centrifuge tube. Place a cap on the tube, and shake vigorously until an emulsion is formed. Loosen the cap, and heat the tube on a steam bath for 10 min. Allow to cool, centrifuge, and determine the pH of the aqueous phase. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Benzyl Alcohol RS USP Ciclopirox Olamine RS RU
Ciclopirox Olamine Topical Suspension

(Comment on this Monograph)id=m17635=Ciclopirox Olamine Topical Suspension=Chl-Cy-Monos.pdf) DEFINITION Ciclopirox Olamine Topical Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of ciclopirox olamine (C12H17NO2 C2H7NO).
USP 32
Temperature Column: 100 Injector: 315 Detector: 315 Carrier gas: Nitrogen Flow rate: 45 mL/min Injection size: 2 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.6 between the peaks Tailing factor: NMT 3.5 for the benzyl alcohol peak and internal standard peak Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution [NOTEAfter six injections, raise the column temperature to 300 for about 5 min, then cool to 100.] Calculate the percentage of benzyl alcohol in the portion of Topical Suspension taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the benzyl alcohol peak to the internal standard peak from the Sample solution = peak response ratio of the benzyl alcohol peak RS to the internal standard peak from the Standard solution CS = concentration of benzyl alcohol in the Standard solution (mg/mL) = nominal concentration of benzyl alcohol in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of the claimed amount of benzyl alcohol RU PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS PH 791: 5.08.0 Analysis: Add 15 mL of boiling water, previously adjusted with 0.1 N hydrochloric acid or 0.1 N sodium hydroxide to a pH of 67, to 3.5 g of Topical Suspension in a 50-mL centrifuge tube. Place a cap on the tube, and shake vigorously until an emulsion is formed. Loosen the cap, and heat the tube on a steam bath for 10 min. Allow to cool, centrifuge, and determine the pH of the aqueous phase. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Benzyl Alcohol RS
Official Monographs / Cilastatin 63

USP Ciclopirox Olamine RS
Cilastatin Sodium
(Comment on this Monograph)id=m17650=Cilastatin Sodium=Chl-Cy-Monos.pdf)
380.44 C16H25N2NaO5S 2-Heptenoic acid, 7-[(2-amino-2-carboxyethyl)thio]-2-[[(2,2dimethylcyclopropyl)carbonyl]amino]-, monosodium salt, [R[R*,S*-(Z)]]-; Sodium (Z)-7-[[(R)-2-amino-2-carboxyethyl]thio]-2-[(S)2,2dimethylcyclopropanecarboxamido]-2-heptenoate [81129-83-1]. DEFINITION Cilastatin Sodium contains NLT 98.0% and NMT 101.5% of C16H25N2NaO5S, calculated on the anhydrous and solvent-free basis. IDENTIFICATION A. The retention time of the major peak for cilastatin from the Sample solution, as obtained in the test for Organic impurities, Procedure 1, corresponds to that of a similar preparation of USP Cilastatin Ammonium Salt RS. B. Ignite a small portion of it on a platinum wire in a nonluminous flame: an intense yellow color is imparted to the flame. ASSAY PROCEDURE Sample solution: Transfer 300 mg of Cilastatin Sodium to a suitable beaker, add 30 mL of methanol, dissolve by swirling, and add 5 mL of water. Analysis: Titrate potentiometrically with 0.1 N hydrochloric acid to a pH of about 3. Then titrate with 0.1 N sodium hydroxide until three inflection points have been observed. Calculate the titer difference, in mL, between the first and third inflection points. Each mL of 0.1 N sodium hydroxide is equivalent to 19.022 mg of C16H25N2NaO5S.
64
Cilastatin / Official Monographs

98.0%101.5%
USP 32
Acceptance criteria Any individual impurity: NMT 0.5% PROCEDURE 2: LIMIT OF SOLVENTS Internal standard solution: 0.5 L/mL of n-propyl alcohol in water Standard stock solution: 2 L/mL of acetone, 0.5 L/mL of methanol, and 0.5 L/mL of mesityl oxide in water. Standard solution: Transfer 2.0 mL of Standard stock solution and 2.0 mL of Internal standard solution to a 10-mL volumetric flask and dilute with water to volume. This solution contains 316 g of acetone/mL, 79 g of methanol/mL, and 86 g of mesityl oxide/mL Sample solution: Transfer 200 mg of Cilastatin Sodium to a 10-mL volumetric flask, add 2.0 mL of Internal standard solution and 5 mL of water, and dissolve by shaking. Dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.53-mm 30-m capillary column, the internal wall of which is coated with a 1.0-m film of liquid phase G16 Temperature Column: See the temperature program table below.
Time (min) 0 2.5 5.0 5.5 Temperature () 50 50 70 70
IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1 Solvent: Water Solution A: Acetonitrile and dilute phosphoric acid (1 in 1000) (3:7). Filter through a 0.5-m or finer porosity. Solution B: Dilute phosphoric acid (1 in 1000). Filter through a 0.5-m or finer porosity. Mobile phase: Use variable mixtures of Solution A and Solution B; see the gradient table below.
Time (min) 0 30 Solution A (%) 15 100 Solution B (%) 85 0
Sample solution: 1.6 mg/mL of Cilastatin Sodium in Solvent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.5-mm 25-cm; packing L1 Temperature: 50 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Sample solution Suitability requirements Capacity factor, k: NLT 10 Column efficiency: NLT 3000 theoretical plates from the cilastatin peak Tailing factor: NMT 4.5 Analysis Samples: Solvent and Sample solution Calculate the chromatographic purity, in percentage, of the Cilastatin Sodium taken: Result = 100 ru/(rT rB rA) = area of the cilastatin peak from the Sample solution = sum of the areas of all the peaks from the rT Sample solution = sum of the areas of all the peaks from the rB Solvent = response of the peak, if any, of nonretained rA substances, such as acetone, at the solvent front from the Sample solution Acceptance criteria: NLT 98.5% Calculate the percentage of each impurity in the portion of Cilastatin Sodium taken: Result = 100 ri/(rT rB rA) ri rT rB rA = peak area for each impurity from the Sample solution = sum of the areas of all the peaks from the Sample solution = sum of the areas of all the peaks from the Solvent = response of the peak, if any, of nonretained substances, such as acetone, at the solvent front from the Sample solution ru
Injector: 160 Detector: 250 Carrier gas: Helium Flow rate: 9 mL/min Injection size: 1 L System suitability Sample: Standard solution Suitability requirements [NOTEThe relative retention times for acetone, methanol, n-propyl alcohol, and mesityl oxide are about 0.26, 0.35, 0.67, and 1.0, respectively.] Relative standard deviation: NMT 5.0% determined from peak area ratios of each analyte to n-propyl alcohol Analysis Samples: Standard solution and Sample solution Inject using the solvent (water) flush technique. Calculate the percentages of acetone, methanol, and mesityl oxide in the Cilastatin Sodium taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = peak area ratio of the corresponding analyte to n-propyl alcohol from the Sample solution = peak area ratio of the corresponding analyte to n-propyl alcohol from the Standard solution = concentration of the appropriate analyte in the Standard solution (g/mL) = concentration of the Sample solution (g/mL)
USP 32
Acceptance criteria Acetone: NMT 1.0% Methanol: NMT 0.5% Mesityl oxide: NMT 0.4% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +41.5 to +44.5, on the anhydrous and solvent-free basis Sample solution: 10 mg/mL, in a mixture of methanol and hydrochloric acid (120:1) PH 791: 6.57.5, in a 10 mg/mL solution BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cilastatin Sodium is sterile, it contains NMT 0.17 USP Endotoxin Unit/mg of cilastatin. STERILITY TESTS 71: Where the label states that Cilastatin Sodium is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration, 6 g of specimen dissolved in 200 mL of Fluid A being used. WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1, and store in a cold place. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile. USP REFERENCE STANDARDS 11 USP Cilastatin Ammonium Salt RS USP Endotoxin RS
Time (min) 0 6.5 10 20 20.1 28
Official Monographs / Cilostazol 65

Solution A (%) 100 50 0 0 100 100 Solution B (%) 0 50 100 100 0 0
Cilostazol
(Comment on this Monograph)id=m17660=Cilostazol=Chl-CyMonos.pdf)
C20H27N5O2 2(1H)-Quinolinone, 6-[4-(1-cyclohexyl-1H-tetrazol-5yl)butoxy]-3,4-dihydro-; 6-[4-(1-Cyclohexyl-1H-tetrazol-5-yl)butoxy]-3,4dihydrocarbostyril [73963-72-1]. DEFINITION Cilostazol contains NLT 98.0% and NMT 102.0% of C20H27N5O2, calculated on the dried basis.
369.46
System suitability solution: 0.05 mg/mL each of USP Cilostazol RS, USP Cilostazol Related Compound A RS, and USP Cilostazol Related Compound B RS in Diluent Standard solution: Prepare 1.0 mg/mL of USP Cilostazol RS in acetonitrile, with sonication if necessary. Transfer 4 mL of this solution to a 10-mL volumetric flask, and dilute with water to volume. Further dilute this solution with Diluent to obtain a solution having a concentration of about 0.04 mg/mL. Sample solution: Transfer 20 mg of Cilostazol to a 50-mL volumetric flask, dissolve in 20 mL of acetonitrile, sonicate if necessary, and dilute with water to volume. Transfer 1 mL of this solution to a 10-mL volumetric flask, and dilute with Diluent to volume (0.04 mg/mL). Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 10-cm; 3.5-m packing L7 Temperature: 40 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution [NOTEFor relative retention times, see Impurity Table 1 under Organic Impurities.] Suitability requirements Resolution: NLT 3.0 between cilostazol related compound B and cilostazol Tailing factor: NMT 2.0 for the cilostazol peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C20H27N5O2 in the portion taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution rU = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221 Sample solution: Dissolve 0.5 g of Cilostazol in 40 mL of dimethylformamide, and add 6 mL of diluted nitric acid and dimethylformamide to make 50 mL. Control solution: To 0.25 mL of 0.01 M hydrochloric acid, add 6 mL of diluted nitric acid and dimethylformamide to make 50 mL. Analysis: Add 1 mL of silver nitrate TS to the Sample solution and to the Control solution, mix well, and allow to stand for 5 min, protecting from direct sunlight. Compare
IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Acetonitrile and water (2:3) Solution A: Acetonitrile and water (3:7) Solution B: Acetonitrile and water (1:1) Mobile phase: Use variable mixtures of Solution A and Solution B.
66
Cilostazol / Official Monographs

Impurity Table 1 Relative Retention Time 0.2 0.9 1.9 Relative Response Factor 1.7 0.58 1.0
USP 32
the opalescence developed in both solutions against a black background by viewing downward or transversely. Acceptance criteria: The opalescence developed in the Sample solution is NMT that of the Control solution (0.018%). HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Diluent, Solution A, Solution B, Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Standard stock solution: 0.5 mg/mL each of USP Cilostazol RS and USP Cilostazol Related Compound C RS in acetonitrile, using sonication if necessary. Transfer 4 mL of Standard stock solution to a 10-mL volumetric flask, and dilute with water to volume. Further dilute this solution, stepwise if necessary, with Diluent to obtain a solution having known concentrations of about 0.4 g/mL of each component. Sample solution: Transfer 20 mg of Cilostazol to a 50-mL volumetric flask, dissolve in 20 mL of acetonitrile, with sonication if necessary, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Analysis Samples: Standard solution and Sample solution Calculate the percentage of cilostazol related compound C taken: Result = (rU/rS) (CS/CU) F 100 = cilostazol related compound C peak response from the Sample solution = cilostazol related compound C peak response rS from the Standard solution = concentration of cilostazol related compound C CS in the Standard solution (g/mL) = concentration of Cilostazol in the Sample CU solution (mg/mL) F = unit conversion factor (g to mg) Calculate the percentage of other impurities: Result = (ri/rS) (CS/CU) F RRF 100 ri rS CS CU F RRF = peak response for any other impurity from the Sample solution = cilostazol peak response from the Standard solution = concentration of Cilostazol in the Standard solution (g/mL) = concentration of Cilostazol in the Sample solution (mg/mL) = unit conversion factor (g to mg) = relative response factor from Impurity Table 1 rU
Name Cilostazol related compound Aa Cilostazol related compound Bb Cilostazol related compound Cc Any other individual impurity
a b
Acceptance Criteria, NMT (%) 0.1 0.1 0.1 0.1
6-Hydroxy-3,4-dihydro-1H-quinolin-2-one. 6-[4-(1-Cyclohexyl-1H-tetrazol-5-yl)-butoxy]-1H-quinolin-2-one. c1-(4-(5-Cyclohexyl-1H-tetrazol-1-yl)butyl)-6-(4-(1-cyclohexyl-1H-tetrazol-5yl)butoxy)-3,4-dihydroquinolin-2(1H)-one.
SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 110 for 3 h: it loses NMT 0.25% of its weight, previously dried at 110 for 3h. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at room temperature. USP REFERENCE STANDARDS 11 USP Cilostazol RS USP Cilostazol Related Compound A RS USP Cilostazol Related Compound B RS USP Cilostazol Related Compound C RS
Cilostazol Tablets
(Comment on this Monograph)id=m17665=Cilostazol Tablets=Chl-Cy-Monos.pdf) DEFINITION Cilostazol Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of cilostazol (C20H27N5O2). IDENTIFICATION A. INFRARED ABSORPTION 197S Standard solution: 100 mg/mL of USP Cilostazol RS in chloroform
USP 32
Sample solution: Equivalent to 100 mg of cilostazol from finely powdered Tablets, into a glass container. Add 1 mL of chloroform, shake for 1 min, and pass through a 0.5-m or finer filter. B. The retention time of the cilostazol peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, and water (7:3:10) Internal standard solution: 4 mg/mL of benzophenone in methanol Standard solution: 0.1 mg/mL of USP Cilostazol RS and 0.04 mg/mL of the Internal standard solution in methanol Sample solution: Equivalent to 50 mg of cilostazol from powdered Tablets (NLT 20), into a suitable volumetric flask and add an appropriate quantity of Internal standard solution. Dilute quantitatively, and stepwise if necessary, with methanol to obtain a solution having a known concentration of about 0.1 mg/mL of USP Cilostazol RS and 0.04 mg/mL of the internal standard. Pass a portion of this solution through a membrane filter having a 0.5-m or finer porosity, and use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 150-cm; packing L1 Temperature: Ambient Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 9.0 between the cilostazol and benzophenone peaks eluted in this order Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of the labeled amount of C20H27N5O2 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cilostazol to the internal standard from the Sample solution = peak response ratio of cilostazol to the internal RS standard from the Standard solution = concentration of USP Cilostazol RS in the CS Standard solution (mg/mL) = nominal concentration of cilostazol in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements
Official Monographs / Cimetidine 67

USP REFERENCE STANDARDS 11 USP Cilostazol RS
Cimetidine
(Comment on this Monograph)id=m17670=Cimetidine=Chl-CyMonos.pdf)
C10H16N6S 252.34 Guanidine, N-cyano-N-methyl-N-[2-[[(5-methyl-1H-imidazol-4yl)methyl]thio]ethyl]-; 2-Cyano-1-methyl-3-[2-[[(5-methylimidazol-4yl)methyl]thio]ethyl]guanidine [51481-61-9]. DEFINITION Cimetidine contains NLT 98.0% and NMT 102.0% of C10H16N6S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The UV absorption spectrum of a solution (1 in 80,000) in 0.1 N sulfuric acid exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Cimetidine RS, concomitantly measured. ASSAY PROCEDURE Mobile phase: Transfer 200 mL of methanol and 0.3 mL of phosphoric acid to a 1000-mL volumetric flask, dilute with water to volume, and filter. Standard stock solution: 0.4 mg/mL of USP Cimetidine RS in methanol and water (1:4) [NOTEPrepare by initially dissolving the USP Cimetidine RS in one part methanol, and diluting that solution with four parts water in an appropriate volumetric flask.] Standard solution: 0.01 mg/mL of USP Cimetidine RS in Mobile phase from the Standard stock solution Sample stock solution: 0.4 mg/mL of Cimetidine in methanol and water, prepared as directed for the Standard stock solution Sample solution: 0.01 mg/mL of Cimetidine in Mobile phase from the Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight and light-resistant containers. Store at controlled room temperature.
68
Cimetidine / Official Monographs

Acceptance criteria Any single impurity: NMT 0.2% Total impurities: NMT 1.0%
USP 32
Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 0.6 Column efficiency: NLT 1000 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H16N6S: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cimetidine RS in the Standard solution (mg/mL) = concentration of Cimetidine in the Sample CU solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% HEAVY METALS, Method II 231: NMT 0.002% Organic Impurities PROCEDURE Mobile phase: Mix 240 mL of methanol, 0.3 mL of 85% phosphoric acid, 940 mg of sodium 1-hexanesulfonate, and sufficient water to make 1 L. Filter before use. Standard solution: 0.80 g/mL of USP Cimetidine RS in Mobile phase Sample solution: 0.4 mg/mL of Cimetidine in Mobile phase [NOTEMix, sonicate for 15 min, and then mix again.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 3.0 Column efficiency: NLT 2000 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the Cimetidine: Result = (ri/rS) (CS/CU) 100 ri rS CS CU = peak response for each impurity from the Sample solution = peak response of the cimetidine from the Standard solution = concentration of USP Cimetidine RS in the Standard solution (g/mL) = concentration of Cimetidine in the Sample solution (g/mL) rU rS CS
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: Between 139 and 144 LOSS ON DRYING 731: Dry a sample at 110 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Cimetidine RS
Cimetidine Tablets
(Comment on this Monograph)id=m17700=Cimetidine Tablets=Chl-Cy-Monos.pdf) DEFINITION Cimetidine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of cimetidine (C10H16N6S). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Transfer 200 mL of methanol and 0.3 mL of phosphoric acid to a 1000-mL volumetric flask, dilute with water to volume, and filter. Standard stock solution: 0.4 mg/mL of USP Cimetidine RS in methanol and water (1:4) [NOTEPrepare by initially dissolving the USP Cimetidine RS in one part methanol and diluting that solution with four parts water in an appropriate volumetric flask.] Standard solution: 0.01 mg/mL of USP Cimetidine RS in Mobile phase from the Standard stock solution Sample stock solution: Transfer an amount of finely powdered Tablets (NLT 20), equivalent to 100 mg of cimetidine, to a 250-mL volumetric flask. Add 50 mL of methanol, shake for 2 min, add 40 mL of water, sonicate for 15 min, and dilute with water to volume. Sample solution: Transfer 5.0 mL of the Sample stock solution to a 200-mL volumetric flask and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 0.6 Column efficiency: NLT 1000 theoretical plates Relative standard deviation: NMT 2.0% for the response for replicate injections
USP 32
Analysis: Calculate the percentage of C10H16N6S in the portion of the Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cimetidine RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 1: 100 rpm [NOTEA 20-mesh basket may be used for 800-mg strength Tablets.] Time: 15 min Detector: UV 218 nm Sample solution: Sample under test. Dilute with Medium to a concentration that is similar to the Standard solution, and filter. Standard solution: USP Cimetidine RS in Medium Tolerances: NLT 80% (Q) of the labeled amount of C10H16N6S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cimetidine RS
Official Monographs / Cimetidine 69

Sample solution: 0.0115 mg/mL of Cimetidine Hydrochloride diluted with Mobile phase from the Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements: Capacity factor: NLT 0.6 Column efficiency: NLT 1000 theoretical plates, determined from the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H16N6SHCl in the portion of the Cimetidine Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Cimetidine Hydrochloride in the Standard solution (mg/mL) CU = concentration of Cimetidine Hydrochloride in the Sample solution (mg/mL) Acceptance criteria: NLT 98.0% and NMT 102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Mobile phase: Transfer 940 mg of sodium 1hexanesulfonate to a 1000-mL volumetric flask, add 240 mL of methanol followed by 0.3 mL of phosphoric acid, and dilute to volume with water. Filter before use. Sample solution 1: 0.4 mg/mL of Cimetidine Hydrochloride in Mobile phase Sample solution 2: 0.8 g/mL of Cimetidine Hydrochloride diluted with Mobile phase from Sample solution 1 System suitability solution: Dissolve 50 mg of Cimetidine Hydrochloride in 10 mL of 1 N hydrochloric acid, heat on a steam bath for 10 min (or boil on a hot plate for 2 min), and allow to cool. Dilute a suitable volume of this solution with Mobile phase to obtain a solution having a concentration of 2 g/mL. [NOTEUse this solution within 24 h of its preparation. Adjusting of the heating step may be necessary to achieve a satisfactory amide analog peak response for the measurement of the resolution between the cimetidine and the amide analog peaks.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: System suitability solution and Sample solution 2 Suitability requirements Resolution: NLT 4.0 between the cimetidine and the amide analog peaks for the System suitability solution rU rS CS
Cimetidine Hydrochloride
(Comment on this Monograph)id=m17730=Cimetidine Hydrochloride=Chl-Cy-Monos.pdf) 288.81 C10H16N6S HCl Guanidine, N-cyano-N-methyl-N-[2-[[(5-methyl-1H-imidazol-4yl)methyl]thio]ethyl]-, monohydrochloride; 2-Cyano-1-methyl-3-[2-[[(5-methylimidazol-4yl)methyl]thio]ethyl]guanidine monohydrochloride [70059-30-2]. DEFINITION Cimetidine Hydrochloride contains NLT 98.0% and NMT 102.0% of C10H16N6S HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Solution: 14 g/mL Medium: 0.1 N sulfuric acid ASSAY PROCEDURE Mobile phase: Transfer 200 mL of methanol and 0.3 mL of phosphoric acid to a 1000-mL volumetric flask, dilute to volume with water, and filter. Standard stock solution: 0.5 mg/mL of USP Cimetidine Hydrochloride RS in a mixture of methanol and water (4:1) Standard solution: 0.0125 mg/mL of USP Cimetidine Hydrochloride RS from the Standard stock solution diluted with Mobile phase Sample stock solution: 0.46 mg/mL Cimetidine Hydrochloride prepared as follows: dissolve 115 mg of Cimetidine Hydrochloride in 50 mL of water in a 250-mL volumetric flask, add 50 mL of methanol, and dilute to volume with water.
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Cimetidine / Official Monographs

Capacity factor: NLT 3.0 for Sample solution 2 Column efficiency: NLT 2000 theoretical plates for Sample solution 2 Relative standard deviation: NMT 7.0% for replicate injections of Sample solution 2 Analysis Samples: Sample solution 1 and Sample solution 2 Calculate the percentage of each impurity in the portion of Cimetidine Hydrochloride taken: Result = (rI/rS) D 100 = peak response for each impurity of the Sample solution 1 = peak response of the cimetidine of the Sample rS solution 2 D = dilution factor to prepare Sample solution 2 from Sample solution 1 Acceptance criteria Any single impurity: NMT 0.2% Total impurities: NMT 1.0% rI
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Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 0.6 Column efficiency: NLT 1000 theoretical plates, determined from the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and the Sample solution Calculate the percentage of C10H16N6S in each mL of the Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cimetidine Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of cimetidine in Sample solution (mg/mL) = molecular weight of cimetidine, 252.34 Mr1 = molecular weight of cimetidine hydrochloride, Mr2 288.81 Acceptance criteria: NLT 90.0% and NMT 110.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.5 USP Endotoxin Units/mg of cimetidine hydrochloride. PH 791: Between 3.8 and 6.0 OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose glass or plastic containers. Glass containers are preferably of Type I or Type II glass. USP REFERENCE STANDARDS 11 USP Cimetidine Hydrochloride RS USP Endotoxin RS rU rS CS
SPECIFIC TESTS LOSS ON DRYING 731 Dry a sample at 105 for 2 h: it loses NMT 0.590% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Cimetidine Hydrochloride RS
Cimetidine Injection
(Comment on this Monograph)id=m17740=Cimetidine Injection=Chl-Cy-Monos.pdf) DEFINITION Cimetidine Injection is a sterile solution of Cimetidine Hydrochloride in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C10H16N6S. IDENTIFICATION The retention time of the major peak for cimetidine of the Sample solution corresponds to that of the cimetidine peak of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Transfer 200 mL of methanol and 0.3 mL of phosphoric acid to a 1000-mL volumetric flask, dilute with water to volume, and filter. Standard stock solution: 0.5 mg/mL of USP Cimetidine Hydrochloride RS in a mixture of methanol and water (80:20) Standard solution: 0.0125 mg/mL of USP Cimetidine Hydrochloride RS in Mobile phase: from the Standard stock solution. Sample solution: Nominally equivalent of 0.02 mg/mL of cimetidine in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
Cimetidine in Sodium Chloride Injection

(Comment on this Monograph)id=m17755=Cimetidine in Sodium Chloride Injection=Chl-Cy-Monos.pdf) DEFINITION Cimetidine in Sodium Chloride Injection is a sterile solution of Cimetidine Hydrochloride and Sodium Chloride in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount C10H16N6S and NLT 95.0 percent and NMT 110.0 percent of the labeled amount of NaCl. IDENTIFICATION A. PROCEDURE: The chromatogram obtained from the Sample solution exhibits a major peak for cimetidine, the
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retention time of which corresponds to that of the cimetidine peak in the chromatogram of the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Sodium 191, and Chloride 191: It meets the requirements. ASSAY CIMETIDINE Mobile phase: Methanol, phosphoric acid, and water (200:0.3:799.7) Standard solution: 0.5 mg/mL of USP Cimetidine Hydrochloride RS in water and methanol (4:1). Dilute in Mobile phase to 0.0125 mg/mL. Sample solution: Equivalent to 0.01 mg/mL of cimetidine from Injection, in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 0.6 Column efficiency: NLT 1000 theoretical plates from the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage label claim of C10H16N6S: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response obtained from the Sample solution = peak response obtained from the Standard solution = concentration of USP Cimetidine Hydrochloride CS RS in the Standard solution (mg/mL) = The nominal concentration of cimetidine in the CU Sample solution (mg/mL) Mr1 = molecular weight ratio of cimetidine (252.34) = molecular weight ratio of cimetidine Mr2 hydrochloride (288.81) Acceptance criteria: 90.0%110.0% SODIUM CHLORIDE Sample solution: 0.5 mg/mL of sodium chloride from Injection in water Analysis: Titrate with 0.1 N silver nitrate VS, using a silversilver chloride electrode. Each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of chloride. From the determined chloride concentration per mL, subtract the quantity, (35.453/252.35)W, to correct for the chloride present as cimetidine hydrochloride, where W = quantity of cimetidine in the Injection (mg/mL), as determined in the Assay for cimetidine. Multiply the corrected value by 1.648 to obtain the quantity, in mg/mL, of sodium chloride in the volume of Injection taken. Acceptance criteria: 95.0%110.0% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: Contains NMT 0.5 USP Endotoxin Units/mg of cimetidine hydrochloride PH 791: 5.07.0 OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose glass or plastic containers. Glass containers are preferably of Type I or Type II glass. rU rS
Official Monographs / Cinoxacin 71

USP REFERENCE STANDARDS 11 USP Cimetidine Hydrochloride RS USP Endotoxin RS
Cinoxacin
(Comment on this Monograph)id=m17830=Cinoxacin=Chl-CyMonos.pdf)
C12H10N2O5 262.22 [1,3]Dioxolo[4,5-g]cinnoline-3-carboxylic acid, 1-ethyl-1,4dihydro-4-oxo-; 1-Ethyl-1,4-dihydro-4-oxo[1,3]dioxolo[4,5-g]cinnoline-3carboxylic acid [28657-80-9]. DEFINITION Cinoxacin contains NLT 97.0% and NMT 102.0% of C12H10N2O5, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution A in the chromatogram prepared as directed in the Procedure under Organic Impurities. ASSAY PROCEDURE Solution A: 38.1 mg/mL of sodium borate in water Internal standard solution: Prepare an aqueous solution containing 2 mg/mL of sulfanilic acid and 5.0 mL of Solution A in each 100 mL. Mobile phase: Dilute 100.0 mL of Solution A and 0.426 g of sodium sulfate with water to 1000 mL, mix, and degas. [NOTEThe quantity of sodium sulfate may be varied to meet System suitability requirements, and to provide a suitable elution time.] Standard stock solution: 1 mg/mL of USP Cinoxacin RS in Solution A Standard solution: Dilute 5.0 mL of Standard stock solution and 5.0 mL of Internal standard solution with water to 100 mL (50 g/mL USP Cinoxacin RS). Sample stock solution: 1 mg/mL of Cinoxacin in Solution A Sample solution: Dilute 5.0 mL of Sample stock solution and 5.0 mL of Internal standard solution with water to 100 mL. Chromatographic system Mode: LC Detector: UV 254 nm Column: 1.8-mm 1-m; packing L12 Flow rate: 1 mL/min Injection size: 1.0 L System suitability Sample: Standard solution [NOTEThe relative retention times for cinoxacin and sulfanilic acid are 1.0 and 2.2, respectively.] Suitability requirements Resolution: NLT 4.4 between cinoxacin and sulfanilic acid Tailing factor: NMT 2.1 for the cinoxacin peak Relative standard deviation: NMT 2.0% from five replicate injections
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Cinoxacin / Official Monographs
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Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H10N2O5 in the portion taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cinoxacin to the peak response of sulfanilic acid from the Sample solution RS = peak response ratio of cinoxacin to the peak response of sulfanilic acid from the Standard solution = concentration of USP Cinoxacin RS in the CS Standard solution (g/mL) = concentration of Cinoxacin in the Sample CU solution (g/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Organic Impurities PROCEDURE Diluent: Chloroform, dimethylformamide, dimethyl sulfoxide, and nitromethane (1:1:1:1) Standard solution A: 5 mg/mL of USP Cinoxacin RS in Diluent Standard solution B: 0.05 mg/mL from Standard solution A in Diluent Sample solution: 5 mg/mL of Cinoxacin in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Acetonitrile, water, and ammonium hydroxide (105:30:7.5) Analysis Samples: Standard solution A, Standard solution B, and Sample solution. Place a volume of the Developing solvent system sufficient to develop the chromatogram, cover, and allow to equilibrate for 30 min. Apply the solutions, dry the plate, and apply each solution three additional times at the corresponding initial locations. Dry the plate thoroughly after each application, and develop the chromatogram until the solvent front has moved to the top of the plate. Remove the plate from the developing chamber, and allow the solvent to evaporate. View the plate under short- and long-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A, and no spot of the Sample solution, other than the principal spot, is larger or more intense than the principal spot of Standard solution B (1.0%). SPECIFIC TESTS LOSS ON DRYING 731: Dry it in vacuum at 60 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cinoxacin RS RU
Cinoxacin Capsules
(Comment on this Monograph)id=m17835=Cinoxacin Capsules=Chl-Cy-Monos.pdf) DEFINITION Cinoxacin Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of C12H10N2O5. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Diluent: Chloroform, dimethylformamide, dimethyl sulfoxide, and nitromethane (1:1:1:1) Standard solution: 5 mg/mL of USP Cinoxacin RS in Diluent Sample solution: Equivalent to 5 mg/mL of cinoxacin from Capsules in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Acetonitrile, water, and ammonium hydroxide (105:30:7.5) Chromatographic chamber: Lined with paper, place a volume of Developing solvent system sufficient to develop the chromatogram, cover, and allow to equilibrate for 30 min. Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Apply the solutions, dry the plate, and apply each solution three additional times at the corresponding initial locations. Dry the plate thoroughly after each application, and develop the chromatogram until the solvent front has moved to the top of the plate. View the plate under short- and long-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Sample solution: Equivalent to 2.5 mg/mL of cinoxacin from the mixed contents of NLT 20 Capsules, in 0.1 M sodium borate. Filter the solution, discarding the first 20 mL of the filtrate, and dilute with water to 10 g/mL. Standard solution: 10 g/mL of USP Cinoxacin RS in the same medium in 0.1 M sodium borate Spectrometric conditions Mode: UV Analytical wavelength: 352 nm Cell: 1 cm Blank: 0.1 M sodium borate and water (1:249) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H10N2O5 in the Capsules: Result = (AU/AS) (CS/CU) 100 AU AS CS = absorbance of the Sample solution = absorbance of the Standard solution = concentration of the Standard solution (g/mL)
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= nominal concentration of the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Buffer solution: pH 6.5 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions) Medium: Buffer solution, 500 mL for Capsules containing 250 mg or less of cinoxacin; 1000 mL for Capsules containing more than 250 mg of cinoxacin Apparatus 1: 100 rpm Time: 30 min Sample solutions: Sample per Dissolution 711. Filter and dilute with 0.1 N sodium hydroxide solution as needed Standard solution: 0.35 mg/mL of USP Cinoxacin RS in Medium Spectrometric conditions Mode: UV Analytical wavelength: 270 nm Analysis Samples: Standard solution and Sample solution Acceptance criteria: NLT 60% (Q) of the labeled amount of C12H10N2O5 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cinoxacin RS CU
Official Monographs / Ciprofloxacin 73

Mode: TLC Adsorbent: 0.25-mm layer of silica gel mixture Application volume: 5 L Analysis Samples: Standard solution and Sample solution Separately apply, as 1-cm bands to the thin-layer chromatographic plate, 5 L of each solution to a suitable thin-layer chromatographic plate. Place the plate in an atmosphere of ammonia for about 15 mins, then transfer the plate to a suitable unsaturated chromatographic chamber, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow the plate to air-dry for about 15 min. Examine the plate under both short- and long-wavelength UV light. Acceptance criteria: The intensity and RF value of the principal band from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Solution A: 0.025 M phosphoric acid. Adjust with triethylamine to a pH of 3.0 0.1. Mobile phase: Acetonitrile and Solution A (13:87) Standard solution: Transfer 12.5 mg of USP Ciprofloxacin RS to a 25-mL volumetric flask. Add 0.1 mL of 7% phosphoric acid and dilute with Mobile phase to volume. System suitability stock solution: 0.025 mg/mL of USP Ciprofloxacin Ethylenediamine Analog RS in Mobile phase System suitability solution: Transfer 1.0 mL of the System suitability stock solution to a 10-mL volumetric flask and dilute with Standard solution to volume. Sample solution: Transfer 25 mg of Ciprofloxacin to a 50mL volumetric flask. Add 0.2 mL of 7% phosphoric acid, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 4.6-mm 25-cm; packing L1 Column temperature: 30 1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for ciprofloxacin ethylenediamine analog and ciprofloxacin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 6 between ciprofloxacin ethylenediamine analog and ciprofloxacin, System suitability solution Column efficiency: NLT 2500 theoretical plates from the ciprofloxacin peak, Standard solution Tailing factor: NMT 2.5 for the ciprofloxacin peak, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Sample: Standard solution and Sample solution. Calculate the percentage of C17H18FN3O3 in the portion of Ciprofloxacin taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
Ciprofloxacin
(Comment on this Monograph)id=m17865=Ciprofloxacin=ChlCy-Monos.pdf)
331.34 C17H18FN3O3 3-Quinolinecarboxylic acid, 1-cyclopropyl-6-fluoro-1,4dihydro-4-oxo-7-(1-piperazinyl)-; 1-Cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-3quinolinecarboxylic acid [85721-33-1]. DEFINITION Ciprofloxacin contains NLT 98.0% and NMT 102.0% of C17H18FN3O3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION The IR absorption spectrum of a potassium bromide dispersion of it exhibits maxima at the same wavelengths as that of a similar preparation of USP Ciprofloxacin RS. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution: 10.0 mg/mL of USP Ciprofloxacin RS in 6 N ammonium hydroxide Sample solution: 10.0 mg/mL of Ciprofloxacin in 6 N ammonium hydroxide Developing solvent system: Methylene chloride, methanol, ammonium hydroxide, and acetonitrile (4:4:2:1) Chromatographic system (See Chromatography 621.)
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= concentration of USP Ciprofloxacin RS in the Standard solution (mg/mL) = concentration of ciprofloxacin in the Sample CU solution (mg/mL) Acceptance criteria: 98.0%102.0% CS
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15 min, transfer the plate to a suitable chromatographic chamber, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow the plate to air-dry for about 15 mins. Examine the plate under short-wavelength UV light. Acceptance criteria: Any spot from the Sample solution, at an RF value corresponding to the principal spot from the Standard solution, is not greater in size or intensity than the principal spot from the Standard solution (0.2%). PROCEDURE 2 Solution A, Mobile phase, System suitability stock solution, System suitability solution, Standard solution, Sample solution, Chromatographic system and System suitability: Prepare as directed in the Assay. Analysis: Proceed as directed in the Assay. Calculate the percentage of each impurity peak in the chromatogram from the Sample solution taken: Result = (rU/rT) 100 rU = peak response of each impurity = sum of the responses of all the peaks rT Acceptance criteria Ciprofloxacin ethylenediamine analog or any other individual impurity peak: NMT 0.2% Total impurities: NMT 0.5% SPECIFIC TESTS CLARITY OF SOLUTION: Dissolve 0.25 g in 10 mL of 0.1 N hydrochloric acid: a clear to slightly opalescent solution is obtained. MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: Where it is intended for use in preparing Ciprofloxacin for Oral Suspension, the total microbial count does not exceed 1000 cfu/g, and the total combined molds and yeast count does not exceed 100 cfu/g. It also meets the requirement for absence of Salmonella species and Escherichia coli. LOSS ON DRYING 731: Dry it in vacuum at 120 for 6 h: it loses NMT 1.0% of its weight, except that where it is labeled as intended for use in preparing Ciprofloxacin for Oral Suspension, it loses between 10% and 20% of its weight. STERILITY TESTS 71: Where the label states that it is sterile, it meets the requirements. BACTERIAL ENDOTOXINS 85: Where the label states that it is sterile or where the label states that Ciprofloxacin must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.50 USP Endotoxin Unit/mg of ciprofloxacin. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at 25, excursion permitted between 15 and 30, and avoid excessive heat. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. Where it is intended for use in preparing Ciprofloxacin for Oral Suspension, it is so labeled. USP REFERENCE STANDARDS 11 USP Ciprofloxacin RS USP Ciprofloxacin Ethylenediamine Analog RS USP Endotoxin RS USP Fluoroquinolonic Acid RS
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1%, except that where it is intended for use in preparing Ciprofloxacin for Oral Suspension, it is NMT 0.2%. CHLORIDE Sample solution: Add 30.0 mL of water to 0.5 g of Ciprofloxacin, shake for 5 min, and filter through chloridefree filter paper. Use the filtrate as the Sample solution. Standard solution: 8.2 g/mL of sodium chloride (5 g/mL of chloride) Samples: Standard solution and Sample solution Analysis Samples: Standard soluton and Sample solution Transfer 15.0 mL of the Sample solution to a 50-mL colorcomparison tube, and transfer 10.0 mL of Standard solution to a second matched 50-mL color-comparison tube, add 5.0 mL of water to the tube containing the Standard solution, and mix. To each tube add 1 mL of 2 N nitric acid, mix, add 1 mL of silver nitrate TS, and mix. Acceptance criteria: The turbidity exhibited by the Sample solution does not exceed that of the Standard solution (0.02%). SULFATE Sample solution: Dissolve 0.5 g in 5.0 mL of 2 N acetic acid and 15.0 mL of water. Standard solution: 18.1 g/mL of potassium sulfate in 30% alcohol (10 g/mL of sulfate) Analysis Samples: Standard solution and Sample solution To each of two 50-mL matched color-comparison tubes transfer 1.50 mL of Standard solution. To each tube add, successively and with continuous shaking, 1.0 mL of 250 mg/mL barium chloride solution, and allow to stand for 1 min. To one of the tubes transfer 15.0 mL of the Standard solution and 0.5 mL of 30% acetic acid, and mix. To the second tube add 15.0 mL of the Sample solution and 0.5 mL of 30% acetic acid, and mix. Acceptance criteria: The turbidity exhibited in the tube containing the Sample solution does not exceed that of the tube containing the Standard solution (0.04%). HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1: LIMIT OF FLUOROQUINOLONIC ACID Standard stock solution: Transfer 5.0 mg of USP Fluoroquinolonic Acid RS to a 50-mL volumetric flask containing 0.05 mL of 6 N ammonium hydroxide and dilute to volume with water. Standard solution: Dilute 2.0 mL of the Standard stock solution to 10.0 mL with water. Sample solution: 10.0 mg/mL of Ciprofloxacin in 0.1 N acetic acid Developing solvent system: Methylene chloride, methanol, acetonitrile and ammonium hydroxide (4:4:1:2) Chromatographic system (See Chromatography 621.) Mode: TLC Adsorbent: 0.25-mm layer of silica gel mixture Application volume: 5 L Analysis Samples: Standard solution and Sample solution Place the plate in a suitable chamber in which is placed a beaker containing 50 mL of ammonium hydroxide. After
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Mode: LC Detector: UV 278 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 30 1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for ciprofloxacin ethylenediamine analog and ciprofloxacin are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 6 between the ciprofloxacin ethylenediamine analog peak and the ciprofloxacin peak, System suitability solution Column efficiency: NLT 2500 theoretical plates from the ciprofloxacin peak, Standard solution Tailing factor: NMT 2.5 for the ciprofloxacin peak, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H18FN3O3 in the portion of Ciprofloxacin Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution rU = peak response of the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Sulfate 221: A 375-mg portion shows no more sulfate than corresponds to 0.15 mL of 0.020 N sulfuric acid (0.04%). HEAVY METALS, Method II 231: NMT 0.002% Organic Impurities PROCEDURE 1: LIMIT OF FLUOROQUINOLONIC ACID Standard solution: Transfer 5.0 mg of USP Fluoroquinolonic Acid RS to a 50-mL volumetric flask containing 0.05 mL of 6 N ammonium hydroxide, add water to volume, and mix. Transfer 2.0 mL of this solution to a 10.0-mL volumetric flask, and dilute with water to volume. Sample solution: 10 mg/mL of Ciprofloxacin Hydrochloride in water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Adsorbent: 0.25-mm layer of silica gel mixture Application volume: 5 L Developing solvent system: Methylene chloride, methanol, acetonitrile, and ammonium hydroxide (4:4:1:2) Analysis Samples: Standard solution and Sample solution Proceed as directed under the General Chapter. Place the plate in a suitable chamber in which is placed a beaker containing 50 mL of ammonium hydroxide. After 15 min, transfer the plate to a suitable chromatographic chamber, and develop the chromatogram in developing solvent system. Allow the chromatogram to develop until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow the plate to air-dry for about 15 min. Examine the plate under short-wavelength UV light.
Ciprofloxacin Hydrochloride
(Comment on this Monograph)id=m17870=Ciprofloxacin Hydrochloride=Chl-Cy-Monos.pdf)
385.82 C17H18FN3O3 HCl H2O 3-Quinolinecarboxylic acid, 1-cyclopropyl-6-fluoro-1,4dihydro-4-oxo-7-(1-piperazinyl)-, monohydrochloride, monohydrate; 1-Cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)-3quinolinecarboxylic acid, monohydrochloride, monohydrate [86393-32-0]. DEFINITION Ciprofloxacin Hydrochloride contains NLT 98.0% and NMT 102.0% of C17H18FN3O3 HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. THIN-LAYER CHROMATOGRAPHY Standard solution: 10 mg/mL of USP Ciprofloxacin Hydrochloride RS in water Sample solution: 10 mg/mL of Ciprofloxacin Hydrochloride in water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of silica gel mixture Application volume: 5 L Developing solvent system: Methylene chloride, methanol, ammonium hydroxide, and acetonitrile (4:4:2:1) Analysis Samples: Standard solution and Sample solution Proceed as directed under the General Chapter. Separately apply, as 1-cm bands to the thin-layer chromatographic plate. Place the plate in an atmosphere of ammonia for about 15 min, then transfer the plate to a suitable unsaturated chromatographic chamber, and develop the chromatogram in Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow the plate to air-dry for about 15 min. Examine the plate under both short- and longwavelength UV light. Acceptance criteria: The intensity and RF value of the principal band of the Sample solution corresponds to that of the Standard solution. C. IDENTIFICATION TESTSGENERAL, Chloride 191 ASSAY PROCEDURE Solution A: 0.025 M phosphoric acid Adjust with triethylamine to a pH of 3.0 0.1. Mobile phase: Acetonitrile and Solution A (13:87) Standard solution: 0.5 mg/mL of USP Ciprofloxacin Hydrochloride RS in Mobile phase System suitability solution: 0.025 mg/mL of USP Ciprofloxacin Ethylenediamine Analog RS in Mobile phase. Transfer 1.0 mL of this solution to a 10-mL volumetric flask, and dilute with Standard solution to volume. Sample solution: 0.5 mg/mL of Ciprofloxacin Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
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Dilute with Mobile phase to 0.13 mg/mL of ciprofloxacin. Standard solution B: Dilute Standard solution with 0.0025 mg/mL of ciprofloxacin from Standard solution A diluted with Mobile phase System suitability solution: 0.04 mg/mL each of USP Ciprofloxacin Hydrochloride RS and USP Ciprofloxacin Ethylenediamine Analog RS in Mobile phase. Dilute 2.0 mL of this solution with Mobile phase to 50 mL. Sample solution: Transfer nominally equivalent to 3 mg of ciprofloxacin from freshly mixed Otic Suspension to a 25-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Samples: Standard solution A, Standard solution B, and System suitability solution Suitability requirements Resolution: Between ciprofloxacin and the ciprofloxacin ethylenediamine analog is NLT 3.0, System suitability solution Column efficiency: NLT 2500 theoretical plates for ciprofloxacin, System suitability solution Tailing factor: NMT 2.0 for ciprofloxacin, from the System suitability solution Relative standard deviation: NMT 2.0% from replicate injections of Standard solution A and Standard solution B Analysis Samples: Standard solution A and Sample solution Calculate the percentage of the label claim of C17H18FN3O3 in the portion of Otic Suspension taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = ciprofloxacin peak response from the Sample solution = ciprofloxacin peak response from the Standard rS solution A = concentration of USP Ciprofloxacin CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of ciprofloxacin in the CU Sample solution (mg/mL) Mr1 = molecular weight of ciprofloxacin, 331.34 = molecular weight of anhydrous ciprofloxacin Mr2 hydrochloride, 367.81 Acceptance criteria: 90.0%110.0% of the labeled amount of C17H18FN3O3 DEXAMETHASONE Solution A and Mobile phase: Prepare as directed under Limit of ciprofloxacin formamide Standard stock solution: 2 mg/mL of USP Dexamethasone RS in acetonitrile. Dilute with Mobile phase Standard solution A: 0.2 mg/mL of USP Dexamethasone RS from Standard stock solution diluted with Mobile phase Standard solution B: 0.004 mg/mL of USP Dexamethazone RS from Standard solution A diluted with Mobile phase System suitability solution: 0.2 mg/mL of USP Dexamethasone RS and 0.2 mg/mL of USP Dexamethasone Acetate RS in Mobile phase Sample solution: Transfer equivalent to 2 mg of dexamethasone from freshly mixed Otic Suspension, to a 10-mL volumetric flask, and dilute with Mobile phase to volume. rU
Acceptance criteria: Any spot of the Sample solution, at an RF value corresponding to the principal spot of the Standard solution, is not greater in size or intensity than the principal spot from the Standard solution (0.2%). PROCEDURE 2 Mobile phase, System suitability solution, Standard solution, Sample solution, Chromatographic system, and System suitability: Prepare as directed in the Assay. Analysis Sample: Sample solution Calculate the percentage of each impurity peak in the chromatogram of the Sample solution taken: Result = (rU/rT) 100 = response of each impurity peak rU = sum of the responses of all the peaks rT Acceptance criteria Individual impurities NMT 0.2% for the ciprofloxacin ethylenediamine analog or any other individual impurity peak Total impurities: NMT 0.5% SPECIFIC TESTS PH 791: 3.04.5, in a 25 mg/mL solution WATER DETERMINATION, Method I 921: 4.7%6.7% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Ciprofloxacin Ethylenediamine Analog RS USP Ciprofloxacin Hydrochloride RS USP Fluoroquinolonic Acid RS
Ciprofloxacin and Dexamethasone Otic Suspension

(Comment on this Monograph)id=m1023=Ciprofloxacin and Dexamethasone Otic Suspension=Chl-Cy-Monos.pdf) DEFINITION Ciprofloxacin and Dexamethasone Otic Suspension is a sterile aqueous suspension containing ciprofloxacin hydrochloride and dexamethasone. It contains NLT 90.0% and NMT 110.0% of the labeled amount of ciprofloxacin (C17H18FN3O3), and NLT 90.0% and NMT 110.0% of the labeled amount of dexamethasone (C22H29FO5). IDENTIFICATION A. The Sample solution, obtained as directed in the Assay for Ciprofloxacin, exhibits a major peak for ciprofloxacin, the retention time of which corresponds to that of the Standard solution, obtained as directed in the Assay for Ciprofloxacin. B. The Sample solution, obtained as directed in the Assay for Dexamethasone, exhibits a major peak for dexamethasone, the retention time of which corresponds to that of the Standard solution, obtained as directed in the Assay for Dexamethasone. ASSAY CIPROFLOXACIN Solution A: Add 6.0 mL of phosphoric acid and 8 g of diethylamine phosphate to 2.0 L of water. Adjust with 50% sodium hydroxide to a pH of 3.0. Mobile phase: Acetonitrile and Solution A (11:89) Standard solution A: 1.48 mg/mL of USP Ciprofloxacin Hydrochloride RS in 0.1 N hydrochloric acid.
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Chromatographic system Mode: LC Detector: UV 254 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1.5 mL/minute Injection size: 50 L System suitability Samples: Standard solution A, Standard solution B, and System suitability solution Suitability requirements Resolution: Between dexamethasone and dexamethasone acetate is NLT 12, System suitability solution Column efficiency: NLT 2000 theoretical plates for dexamethasone, System suitability solution Tailing factor: NMT 2.0 for dexamethasone, System suitability solution Relative standard deviation: NMT 2.0% from replicate injections of the Standard solution A and Standard solution B Analysis Samples: Standard solution and Sample solution Calculate the percentage of the label claim of C22H29FO5 in the portion of Otic Suspension taken: Result = (rU/rS) (CS/CU) 100 = dexamethasone peak response from the Sample solution = dexamethasone peak response from the rS Standard solution CS = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) CU = nominal concentration of dexamethasone in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amount of C22H29FO5 IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF CIPROFLOXACIN FORMAMIDE Solution A: phosphoric acid and water (3:997) Adjust with 50% sodium hydroxide to a pH of 3.0. Mobile phase: Acetonitrile and Solution A (27:73) Standard stock solution: 0.25 mg/mL of USP Ciprofloxacin Formamide RS in methanol Standard solution: 0.015 mg/mL from Standard stock solution diluted with Mobile phase System suitability solution: 0.025 mg/mL of USP Dexamethasone RS and 0.025 mg/mL of USP Ciprofloxacin Formamide RS in methanol and Mobile phase (3:17) [NOTEDissolve in methanol, and dilute with Mobile phase to volume.] Sample solution: Transfer an amount of freshly mixed Otic Suspension, nominal equivalent to 6 mg, to a 10-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1.5 mL/minute Injection size: 50 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: Between ciprofloxacin formamide and dexamethasone is NLT 8, System suitability solution Column efficiency: NLT 2000 theoretical plates for ciprofloxacin formamide, Standard solution Tailing factor: NMT 2.0 for ciprofloxacin formamide, Standard solution rU

Relative standard deviation: NMT 2.0% determined from replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of each related compound in the portion of Otic Suspension taken: Result = (rU/rS) (CS/CU) 100 = ciprofloxacin formamide peak response from the Sample solution = ciprofloxacin formamide peak response from the rS Standard solution = concentration of USP Ciprofloxacin Formamide CS RS in the Standard solution (mg/mL) = nominal concentration of ciprofloxacin of the CU Sample solution (mg/mL) Acceptance criteria: NMT 0.5% of the labeled amount of ciprofloxacin PROCEDURE 2: CIPROFLOXACIN RELATED COMPOUNDS Analysis: From the chromatogram of the Sample solution B, obtained as directed in the Assay for ciprofloxacin, measure the responses for the ciprofloxacin ethylenediamine analog and the other minor peaks. Calculate the percentage of each related compound in the portion of Otic Suspension taken: rU Result = (rU/rS) (CS/CU) (Mr1/Mr2) (100/F) = related compound peak responses from the Sample solution = ciprofloxacin peak response from the Dilute rS Standard solution = concentration of USP Ciprofloxacin CS Hydrochloride RS in the Standard solution B (mg/mL), calculated on the anhydrous basis CU = nominal concentration of ciprofloxacin of the Otic Suspension (mg/mL) = molecular weight of ciprofloxacin, 331.34 Mr1 Mr2 = molecular weight of anhydrous ciprofloxacin hydrochloride, 367.81 F = relative response factor (1.3 for ciprofloxacin ethylenediamine analog and 1.0 assumed for all other degradation products) Acceptance criteria Ciprofloxacin ethylenediamine analog: NMT 0.4% of the labeled amount of ciprofloxacin Other single related compound: NMT 0.2% Sum of all related compounds: NMT 0.8% PROCEDURE 3: DEXAMETHASONE RELATED COMPOUNDS Analysis: From the chromatogram of the Sample solution, obtained as directed in the Assay for dexamethasone, measure the responses for the 21-dehydro-17-deoxy related compound, the 20-carboxy-17-desoxy related compound, and other minor peaks. Calculate the percentage of the label claim of each related compound in the the Otic Suspension taken: rU Result = (rU/rS) (CS/CU) 100 rU rS CS CU = related compound peak responses from the Sample solution = dexamethasone peak response from the Dilute Standard solution = concentration of USP Dexamethasone RS in the Standard solution B (mg/mL) = nominal concentration of dexamethazone of the Otic Suspension (mg/mL)
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Acceptance criteria 21-Dehydro-17-deoxy related compound: NMT 1.0% 20-Carboxy-17-desoxy related compound: NMT 2.6% Other single related compound: NMT 0.3% Sum of all related compounds: NMT 3.5% [NOTEThe relative retention times are about 1.4 to 1.6 for the 21-dehydro-17-deoxy related compound and about 2.8 to 3.2 for the 20-carboxy-17-desoxy related compound.]
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Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Separately apply, as 1-cm bands to the thin-layer chromatographic plate. Place the plate in an atmosphere of ammonia for about 15 min, then transfer the plate to a suitable unsaturated chromatographic chamber, and develop the chromatogram in the Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow the plate to air-dry for about 15 min. Examine the plate under both short- and longwavelength UV light. Acceptance criteria: The intensity and RF value of the principal band from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Solution A: 0.025 M phosphoric acid. Adjust with triethylamine to a pH of 3.0 0.1. Mobile phase: Acetonitrile and Solution A (13:87) (see Chromatography 621, System Suitability) Standard solution: 0.5 mg/mL of USP Ciprofloxacin Hydrochloride RS in Mobile phase System suitability solution: 0.025 mg/mL of USP Ciprofloxacin Ethylenediamine Analog RS in Mobile phase. Transfer 1.0 mL of this solution to a 10-mL volumetric flask, and dilute with Standard solution to volume. Sample solution: Equivalent to 0.5 mg/mL of Ciprofloxacin from Injection diluted with Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 30 1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution and System suitability solution [NOTEThe relative retention times for ciprofloxacin ethylenediamine analog and ciprofloxacin are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 6 between the ciprofloxacin ethylenediamine analog peak and the ciprofloxacin peak Column efficiency: NLT 2500 theoretical plates from the ciprofloxacin peak, Standard solution Tailing factor: NMT 2.5 for the ciprofloxacin peak, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percent label claim of ciprofloxacin in the Injection: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 Mr2 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ciprofloxacin Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of ciprofloxacin, Sample solution (mg/mL) = molecular weight of ciprofloxacin, 331.34 = molecular weight of anhydrous ciprofloxacin hydrochloride, 367.81
SPECIFIC TESTS PH 791: 3.84.8 STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration. PARTICLE SIZE Carrier fluid: Heat Purified Water to a temperature of 40 to 50, add 100 mg/L of dexamethasone while stirring, cool to room temperature while stirring, pass through a 0.2-m filter, and store in a clean, covered container. Sample solution: Dilute a volume of 10 L of Otic Suspension with Carrier fluid to 25 mL. Analysis (See Particulate Matter in Injections 788, Light Obscuration Particle Count Test.) Analyze the Sample solution using an electronic, liquid-borne particle counting system that employs a light obscuration sensor with a suitable sample feeding device. Acceptance criteria: NLT 99.5% of the particles are 25 m, not less than 99.95% are 50 m, and not less than 99.995% are 100 m. OSMOLALITY 785: 270330 mOsmol/kg ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. Avoid freezing. USP REFERENCE STANDARDS 11 USP Ciprofloxacin Ethylenediamine Analog RS USP Ciprofloxacin Formamide RS USP Ciprofloxacin Hydrochloride RS USP Dexamethasone RS USP Dexamethasone Acetate RS
Ciprofloxacin Injection
(Comment on this Monograph)id=m17872=Ciprofloxacin Injection=Chl-Cy-Monos.pdf) DEFINITION Ciprofloxacin Injection is a sterile solution of Ciprofloxacin or Ciprofloxacin Hydrochloride in Water for Injection, in 5% Dextrose Injection, or in 0.9% Sodium Chloride Injection prepared with the aid of Lactic Acid. It contains NLT 90.0% and NMT 110.0% of the labeled amount of ciprofloxacin (C17H18FN3O3). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 0.5 mg/mL of USP Ciprofloxacin Hydrochloride RS in water Sample solution: 0.5 mg/mL of Ciprofloxacin from Injection in water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Methylene chloride, methanol, ammonium hydroxide, and acetonitrile (4:4:2:1)
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Acceptance criteria: 90.0%110.0% of C17H18FN3O3

potassium chromate TS, and titrate with 0.1 N silver nitrate TS. Each mL of 0.1 N silver nitrate is equivalent to 5.844 mg of sodium chloride (NaCl). Acceptance criteria: 85.594.5 mg IMPURITIES Organic Impurities PROCEDURE: LIMIT OF CIPROFLOXACIN ETHYLENEDIAMINE ANALOG Mobile phase, System suitability solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Analysis Samples: Sample solution Proceed as directed for Analysis in the Assay. Calculate the percentage of ciprofloxacin ethylenediamine analog from the chromatogram obtained from the Sample solution: Result = F rA/(0.7rA + rC) 100 = correction factor for ciprofloxacin ethylenediamine analog, 0.7 = ciprofloxacin ethylenediamine analog peak rA response = ciprofloxacin peak response rC Acceptance criteria: NMT 0.5% SPECIFIC TESTS PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements PH 791: 3.54.6, except that where the Injection is labeled as being a concentrated form, its pH is 3.33.9 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.50 USP Endotoxin Unit/mg of ciprofloxacin. STERILITY TESTS 71: It meets the requirements for Test for Sterility of the Product to Be Examined, Membrane Filtration. COLOR AND ACHROMICITY 631 (WHERE IT IS LABELED AS BEING A CONCENTRATED FORM): It has no more color than a solution prepared by diluting 5.0 mL of Matching Fluid O with 95.0 mL of 0.12 N hydrochloric acid. OTHER REQUIREMENTS: It meets the requirements for Injections 1, Volume in Container. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass, and store in a cool place or at controlled room temperature. Avoid freezing and exposure to light. LABELING: The label indicates whether the vehicle is Sterile Water for Injection, 5% Dextrose Injection, or 0.9% Sodium Chloride Injection. Label the Injection that has Sterile Water for Injection as the vehicle to indicate that it is a concentrated form that must be diluted to appropriate strength (12 mg/mL) with 5% Dextrose Injection or 0.9% Sodium Chloride Injection before administration, and that the resulting solution is stable for up to 14 days when stored in a cool place or at controlled room temperature. USP REFERENCE STANDARDS 11 USP Ciprofloxacin Ethylenediamine Analog RS USP Ciprofloxacin Hydrochloride RS USP Endotoxin RS USP Sodium Lactate RS F
OTHER COMPONENTS LACTIC ACID CONTENT Mobile phase: Acetonitrile and 0.005 N sulfuric acid (3:17) Standard solution: 0.8 mg/mL of USP Sodium Lactate RS in water or 4 mg/mL where the Injection is labeled as being a concentrated form Sample solution: Use the undiluted Injection. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 208 nm Column: 7.8-mm 30-cm; packing L17 Temperature: 40 1 Flow rate: 0.6 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2.0% [NOTEAfter each analysis, rinse the column with a mixture of 0.01 N sulfuric acid and acetonitrile to elute the ciprofloxacin from the column. Promptly regenerate the column with 0.01 N sulfuric acid, and the column may be reused or stored.] Analysis Samples: Standard solution and Sample solution Calculate the concentration of lactic acid (C3H6O3) in mg/mg of ciprofloxacin: Result = (rU/rS) (CS/CU) (Mr1/Mr2) = peak response of lactic acid from the Sample solution rS = peak response of lactic acid from the Standard solution = concentration of USP Sodium Lactate RS in the CS Standard solution (mg/mL) CU = nominal concentration of ciprofloxacin in the Sample solution (mg/mL) = molecular weight of lactic acid, 90.08 Mr1 = molecular weight of sodium lactate, 112.07 Mr2 Acceptance criteria: 0.2880.352 mg of lactic acid for each mg of ciprofloxacin claimed on the label, except that where the Injection is labeled as being a concentrated form, it contains 0.3350.409 mg of lactic acid for each mg of ciprofloxacin claimed on the label. DEXTROSE CONTENT (IF PRESENT) Analysis Sample solution: Undiluted Injection Determine the angular rotation in a suitable polarimeter tube (see Optical Rotation 781). Calculate the percentage (g/100 mL) of dextrose (C6H12O6 H2O) in the portion of Injection taken: A R (Mr1/Mr2) (100/F) = 100 mm divided by the length of the polarimeter tube (mm) R = observed rotation (degrees) = molecular weight of dextrose monohydrate, Mr1 198.17 Mr2 = molecular weight of anhydrous dextrose, 180.16 F = midpoint of the specific rotation range for anhydrous dextrose (degrees), 52.9 Acceptance criteria: 4.755.25 g/100 mL SODIUM CHLORIDE CONTENT (IF PRESENT) Sample solution: Injection Analysis: Transfer 10.0 mL of Sample solution to a suitable container, dilute with water to 150 mL, add 1.5 mL of A rU
Ciprofloxacin Ophthalmic Ointment

(Comment on this Monograph)id=m17875=Ciprofloxacin Ophthalmic Ointment=Chl-Cy-Monos.pdf) DEFINITION Ciprofloxacin Ophthalmic Ointment contains an amount of Ciprofloxacin Hydrochloride equivalent to NLT 90.0% and
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SPECIFIC TESTS STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration. METAL PARTICLES IN OPHTHALMIC OINTMENTS 751: Meets the requirements PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. Store at a temperature between 2 and 25. USP REFERENCE STANDARDS 11 USP Ciprofloxacin Ethylenediamine Analog RS USP Ciprofloxacin Hydrochloride RS
NMT 110.0% of the labeled amount of ciprofloxacin (C17H18FN3O3). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.005 M tetrabutylammonium phosphate solution. Adjust with phosphoric acid to a pH of 2.0. Mobile phase: Methanol and Solution A (1:3) System suitability solution: 0.005 mg/mL of USP Ciprofloxacin Ethylenediamine Analog RS in Standard solution Standard solution: 0.033 mg/mL of USP Ciprofloxacin Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: Transfer an amount nominally equivalent to 750 g of ciprofloxacin from Ophthalmic Ointment, to a screw-capped tube. Add 15 mL of solvent hexane, and shake vigorously until the Ophthalmic Ointment is dispersed. Loosen the cap, and heat in a water bath at 60 for 30 min, with occasional swirling. Remove from the bath, tighten the cap, and shake for 1.5 min while still hot. Add 25.0 mL of 0.1 N hydrochloric acid, and shake vigorously for 1.5 min. Allow the layers to separate, and use the lower, aqueous layer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution and System suitability solution [NOTERelative retention times for the ciprofloxacin ethylenediamine analog and ciprofloxacin are 0.8 and 1.0, respectively.] Suitability requirements Resolution: Between ciprofloxacin ethylenediamine analog and ciprofloxacin is NLT 2.0, System suitability solution Column efficiency: NLT 500 theoretical plates, Standard solution Tailing factor: 0.92.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of the label claim of C17H18FN3O3 in the portion of Ophthalmic Ointment taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ciprofloxacin Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of ciprofloxacin in the CU Sample solution (mg/mL) = molecular weight of ciprofloxacin, 331.34 Mr1 = molecular weight of ciprofloxacin hydrochloride Mr2 monohydrate, 385.82 Acceptance criteria: 90.0%110.0% of the labeled amount of C17H18FN3O3 rU rS CS
Ciprofloxacin Ophthalmic Solution

(Comment on this Monograph)id=m17876=Ciprofloxacin Ophthalmic Solution=Chl-Cy-Monos.pdf) DEFINITION Ciprofloxacin Ophthalmic Solution is a sterile, aqueous solution of Ciprofloxacin Hydrochloride. It contains NLT 90.0% and NMT 110.0% of the labeled amount of ciprofloxacin (C17H18FN3O3). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 3.5 mg/mL of USP Ciprofloxacin Hydrochloride RS in water Sample solution: 3 mg/mL of ciprofloxacin from Ophthalmic Solution in water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of silica gel mixture Application volume: 3 L Developing solvent system: Methylene chloride, methanol, ammonium hydroxide, and acetonitrile (4:4:2:1) Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Separately apply, as 1-cm bands to the thin-layer chromatographic plate. Place the plate in an atmosphere of ammonia for about 15 min, then transfer the plate to a suitable unsaturated chromatographic chamber, and develop the chromatogram in developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow the plate to air-dry for about 15 min. Examine the plate under both short- and longwavelength UV light. Acceptance criteria: The intensity and RF value of the principal band from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Solution A: 0.005 M tetrabutylammonium phosphate solution Adjust with phosphoric acid to a pH of 2.0. Mobile phase: Methanol and Solution A (1:3) Standard solution: 0.14 mg/mL of USP Ciprofloxacin Hydrochloride RS in water
USP 32
System suitability solution: 0.01 mg/mL of USP Ciprofloxacin Ethylenediamine Analog RS in Standard solution Sample solution: Equivalent to 0.12 mg/mL of ciprofloxacin from Ophthalmic Solution, in water. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution and System suitability solution [NOTERelative retention times for the ciprofloxacin ethylenediamine analog and ciprofloxacin are 0.8 and 1.0, respectively.] Suitability requirements Resolution: Between ciprofloxacin ethylenediamine analog and ciprofloxacin is NLT 1.5., System suitability solution Capacity factor: 1.56 for the ciprofloxacin peak, Standard solution Column efficiency: NLT 500 theoretical plates, Standard solution Tailing factor: 0.92.0, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of the label claim of C17H18FN3O3 in the portion of Ophthalmic Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ciprofloxacin Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of ciprofloxacin in the CU Sample solution (unit/mL) = molecular weight of ciprofloxacin, 331.34 Mr1 = molecular weight of anhydrous ciprofloxacin Mr2 hydrochloride, 367.81 Acceptance criteria: 90.0%110.0% of the labeled amount of C17H18FN3O3 rU rS CS SPECIFIC TESTS PH 791: 3.55.5 STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers protected from light, at room temperature. USP REFERENCE STANDARDS 11 USP Ciprofloxacin Ethylenediamine Analog RS USP Ciprofloxacin Hydrochloride RS

IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHY Standard solution: 1.5 mg/mL of USP Ciprofloxacin Hydrochloride RS in water Sample solution: Place a number of Tablets, equivalent to about 1500 mg of ciprofloxacin, in a suitable flask containing 750 mL of water, and sonicate for about 20 min. Dilute with water to 1000 mL, and mix. Centrifuge a portion of this suspension, and use the clear supernatant. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of silica gel mixture Application volume: 10 L Developing solvent system: Methylene chloride, methanol, ammonium hydroxide, and acetonitrile (4:4:2:1) Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Separately apply, as 1-cm bands to the thin-layer chromatographic plate. Place the plate in an atmosphere of ammonia for about 15 min, then transfer the plate to a suitable unsaturated chromatographic chamber, and develop the chromatogram in developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow the plate to air-dry for about 15 min. Examine the plate under both short- and longwavelength UV light. Acceptance criteria: The intensity and RF value of the principal band from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Solution A: 0.025 M phosphoric acid. Adjust with triethylamine to a pH of 2.0 0.1. Solution B: Acetonitrile and Solution A (13:87) Solution C: 0.025 M phosphoric acid Adjust with triethylamine to a pH of 3.0 0.1. Mobile phase: Acetonitrile and Solution C (13:87) System suitability solution: 0.05 mg/mL of USP Ciprofloxacin Ethylenediamine Analog RS in Standard solution Standard solution: 0.2 mg/mL of USP Ciprofloxacin Hydrochloride RS in Solution B Sample solution: Transfer 5 Tablets to a 500-mL volumetric flask, add 400 mL of Solution B, and sonicate for about 20 min. Dilute with Solution B to volume, mix and filter through a 0.45 m membrane filter. Prepare nominally 0.20 mg/mL of ciprofloxacin from the filtrate with Solution B. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 278 nm Column: 4.6-mm 25-cm; packing L1 Temperature: 30 1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution and System suitability solution [NOTEThe retention time is 6.410.8 min for ciprofloxacin. Relative retention times for ciprofloxacin ethylenediamine analog and ciprofloxacin are 0.7 and 1.0, respectively.]
Ciprofloxacin Tablets
(Comment on this Monograph)id=m17880=Ciprofloxacin Tablets=Chl-Cy-Monos.pdf) DEFINITION Ciprofloxacin Tablets contain Ciprofloxacin Hydrochloride equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of ciprofloxacin (C17H18FN3O3).
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USP 32
DEFINITION Cisplatin contains NLT 98.0% and NMT 102.0% of Cl2H6N2Pt, calculated on the anhydrous basis. [CAUTIONCisplatin is potentially cytotoxic. Great care should be taken to prevent inhaling particles and exposing the skin to it.] IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. INFRARED ABSORPTION 197K C. THIN-LAYER CHROMATOGRAPHY Standard solution: 1 mg/mL of USP Cisplatin RS in dimethylformamide Sample solution: 1 mg/mL of Cisplatin in dimethylformamide Solution A: 5.6 g of stannous chloride in 10 mL of hydrochloric acid. Stir for 5 min. [NOTEIt is not necessary that all of the solids dissolve.] Solution B: 0.2 g of potassium iodide in 90 mL of water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Acetone and 1 N nitric acid (9:1) Spray reagent A: Mix Solution A and Solution B together. Disregard any precipitate that is formed. [NOTEStore in the dark. The solution is usable for at least 1 week.] Spray reagent B: 20 mg/mL of potassium iodide in water Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Place the plate in a suitable chromatographic chamber containing a filter paper lining and equilibrated for 30 min with the developing solvent system. Develop the plate for a distance of about 8 cm from the origin. Remove the plate, and allow it to air-dry. Complete the drying by heating in a forced-air oven at about 100 for 1 min. Spray the plate with Spray reagent A, heat it in an oven at about 100 for 5 min, cool, and spray with Spray reagent B, to bring out the full color of the spots. Acceptance criteria: The principal spot from the Sample solution corresponds in appearance and RF value to that produced by the Standard solution. ASSAY PROCEDURE Mobile phase: Ethyl acetate, methanol, dimethylformamide, and degassed water (25:16:5:5) Standard solution: 1 mg/mL of USP Cisplatin RS in dimethylformamide [NOTEUse within 1 h.] Sample solution: 1 mg/mL of Cisplatin in dimethylformamide Chromatographic system (See Chromatography 621, System Suitability.)
Suitability requirements Resolution: Between the ciprofloxacin ethylenediamine analog peak and the ciprofloxacin peak is NLT 6, System suitability requirements Column efficiency: NLT 2500 theoretical plates from the ciprofloxacin peak, Standard solution Tailing factor: NMT 2.0 for the ciprofloxacin peak, Standard solution Relative standard deviation: NMT 1.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of the label claim of C17H18FN3O3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) (100 / L) = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Ciprofloxacin Hydrochloride RS in the Standard solution (mg/mL), calculated on the anhydrous basis = nominal concentration of ciprofloxacin in the CU Sample solution (mg/mL) = molecular weight of ciprofloxacin, 331.34 Mr1 = molecular weight of anhydrous ciprofloxacin Mr2 hydrochloride, 367.81 Acceptance criteria: 90.0%110.0% of the labeled amount of C17H18FN3O3 PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Sample solutions: Sample per 711 Dissolution. Dilute with Medium as needed. Standard solution: USP Ciprofloxacin Hydrochloride RS in Medium Spectrometric conditions Mode: UV Analytical wavelength: 276 nm Analysis Samples: Standard solution and Sample solution Tolerances: An amount of ciprofloxacin hydrochloride (C17H18FN3O3 HCl) equivalent to NLT 80% (Q) of the labeled amount of ciprofloxacin (C17H18FN3O3) is dissolved in 30 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Ciprofloxacin Ethylenediamine Analog RS USP Ciprofloxacin Hydrochloride RS rU rS CS
Cisplatin
(Comment on this Monograph)id=m17910=Cisplatin=Chl-CyMonos.pdf)
Cl2H6N2Pt Platinum, diamminedichloro-, (SP-4-2)-; cis-Diamminedichloroplatinum [15663-27-1].
300.04
USP 32
Mode: LC Detector: UV 310 nm Column: 4.0-mm 30-cm; packing L8 Flow rate: 2.0 mL/minute Injection size: 40 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of Cl2H6N2Pt in the portion of Cisplatin taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution rU = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% IMPURITIES Organic Impurities PROCEDURE 1: LIMIT OF TRICHLOROAMMINEPLATINATE Mobile phase: 0.4 mg/mL of ammonium sulfate in water. The pH of this solution is 5.9 0.1. Make adjustments to the ionic strength of the Mobile phase, if necessary, to meet the system suitability requirements. Standard solution: 6 g/mL of USP Potassium Trichloroammineplatinate RS in saline TS [NOTEUse low-actinic glassware.] [NOTEUse within 4 h.] Sample solution: Nominally 0.5 mg/mL of Cisplatin in saline TS [NOTEUse low-actinic glassware.] [NOTECompletely dissolve by stirring by mechanical means for 30 min. Use within 4 h.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 209 nm Column: 4.6-mm 25-cm; packing L14 Flow rate: 2 mL/min Injection size: 20 L System suitability [NOTEThe relative retention times are about 1.0 for cisplatin (in the void volume) and 5.0 for trichloroammineplatinate.] Sample: Standard solution Suitability requirements Resolution: Between the saline TS peak and the trichloroammineplatinate peak is NLT 2.0. Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of trichloroammineplatinate in the portion of Cisplatin taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 Mr2 = = = = peak area from the Sample solution peak area from the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (g/mL) = molecular weight of trichloroammineplatinate, 318.48 = molecular weight of potassium trichloroammineplatinate, 357.58
Official Monographs / Cisplatin 83

Acceptance criteria: NMT 1.0% PROCEDURE 2: LIMIT OF TRANSPLATIN Mobile phase: 0.18 M monobasic potassium phosphate solution. Adjust with phosphoric acid to a pH of 3.2. Standard stock solution: 0.05 mg/mL of USP Transplatin RS in saline TS [NOTEdissolve by stirring by mechanical means for 30 min.] Standard solution A: Add 5 mL of Standard stock solution to 12 mg of USP Cisplatin RS. Dilute with saline TS to 25 mL, and stir by mechanical means for 30 min to dissolve. Standard solution B: Pipet 10 mL of Standard solution A into a 50-mL volumetric flask. Add 5.0 mL of 1 in 200 solution of thiourea (prepared fresh daily) and 5.0 mL of 1 N hydrochloric acid, and dilute with saline TS to volume. Place 10 mL of this solution in a suitable serum vial, seal with a polytef-lined closure, and heat in a heating block at 60 0.5 for 60 min. Remove, and cool to room temperature. Sample solution A: 0.5 mg/mL of Cisplatin in saline TS [NOTEDissolve by stirring by mechanical means for 30 min.] Sample solution B: Pipet 10 mL of Sample solution A into a 50-mL volumetric flask and proceed as directed for Standard solution B, beginning with add 5.0 mL of a 1 in 200 solution of thiourea. System suitability solution: 0.05 mg/mL of USP Cisplatin RS in saline TS. Stir by mechanical means for 30 min to dissolve. Pipet 10 mL of this solution and 10 mL of Standard stock solution into a 50-mL volumetric flask, and proceed as directed for Standard solution B beginning with add 5.0 mL of a 1 in 200 solution of thiourea. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L9 Column temperature: 45 Flow rate: 2 mL/min Injection size: 20 L [NOTECondition the column by pumping Mobile phase at a flow rate of 2 mL/min for 30 min, then at 0.5 mL/min for 30 min, and then again at 2 mL/min for 30 min.] System suitability Samples: Standard solution B and System suitability solution [NOTEThe retention time of the derivatized transplatin is between 5.0 and 9.0 min; or, if it is not, modify the Mobile phase as necessary, and recondition the column. The relative retention times are about 1.0 for cisplatin and 1.3 for transplatin.] Suitability requirements Column efficiency, n: NLT 2500 theoretical plates Relative standard deviation: NMT 4.0% Resolution: NLT 1.7 Analysis Samples: Standard solution B and Sample solution B Calculate the percentage of transplatin in the portion of Cisplatin taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak area from Sample solution B = peak area from Standard solution B = concentration of USP Transplatin RS in Standard solution A (g/mL) = nominal concentration of Cisplatin in the Sample solution A (g/mL)
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Cisplatin / Official Monographs
USP 32
Acceptance criteria: NMT 2.0% UV PURITY RATIO [NOTECleanse all glassware with a mixture of hydrochloric acid and nitric acid (3:1), rinse thoroughly with water, and dry before use. Do not use dichromate for cleaning. Do not use acetone or pressurized air for drying. Protect the Sample solution from light, and use within 1 h after its preparation.] Analysis Sample: 98.5 0.5 mg of ground cisplatin Transfer Sample to a 100-mL volumetric flask, and add 0.1 N hydrochloric acid to volume. Using a clean magnetic stir bar, alternately stir at a high speed for 5 min and sonicate for 10 s until complete solution is effected, inverting the flask frequently to remove particles that may cling to the neck. Obtain the UV absorption spectrum, using thoroughly rinsed 2-cm cells, with 0.1 N hydrochloric acid in the reference cell. Acceptance criteria: The ratio of the absorbance at the maximum near 301 nm to that at the minimum near 246 nm is NLT 4.5. SPECIFIC TESTS PLATINUM CONTENT [NOTEThoroughly cleanse all glassware with nitric acid, and rinse with Purified Water, to prevent mirroring of the platinum precipitate.] Analysis: Transfer 0.5 g of Cisplatin to a 600-mL beaker. Add 300 mL of 0.1 N hydrochloric acid, and slowly dissolve by heating nearly to boiling on a hot plate covered with an insulating pad, and stirring frequently with a glass stirring rod. When solution is complete, remove the insulating pad, and boil for about 10 min. Remove the beaker from the hot plate, allow to cool for 1 min without stirring, and filter through quantitative, fine-porosity, smooth, dense, ashless filter paper, collecting the filtrate in a 600-mL beaker, completing the transfer to the filter with hot water. Wash the filter with hot water. Place the beaker containing the combined filtrate and washings on a hot plate, and evaporate to a volume of 300 mL. Place a glass stirring rod in the beaker, and heat the solution to boiling. Slowly add to the center of the beaker, by dropwise additions, 10.0 mL of hydrazine hydrate, 85%. [CautionHydrazine is toxic.] Add 2 drops of 10 N sodium hydroxide, boil for 10 min to coagulate the precipitate for ease of filtration, cool, and filter through quantitative, medium-porosity, smooth, ashless filter paper. Rinse the beaker with hot water, and pour the rinsings onto the filter. Wipe the beaker and the stirring rod with small pieces of the same kind of paper used for this filtration, and place these and the filter containing the precipitate in a No. 1 porcelain crucible, previously ignited to constant weight. Dry on a hot plate covered with an insulating pad, slowly increase the heat to char, and ignite for 1 h at 800. Cool in a desiccator, and again weigh. Acceptance criteria: The weight of the platinum so obtained is between 64.42% and 65.22% of the weight of Cisplatin taken, on the anhydrous basis. CRYSTALLINITY 695: Meets the requirements WATER DETERMINATION, Method I 921: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Protect from light. USP REFERENCE STANDARDS 11 USP Cisplatin RS USP Transplatin RS USP Potassium Trichloroammineplatinate RS
Cisplatin for Injection

(Comment on this Monograph)id=m17920=Cisplatin for Injection=Chl-Cy-Monos.pdf) DEFINITION Cisplatin for Injection is a sterile, lyophilized mixture of Cisplatin, Mannitol, and Sodium Chloride. It contains NLT 90.0% and NMT 110.0% of the labeled amount of cisplatin (Cl2H6N2Pt). [CAUTIONCisplatin is potentially cytotoxic. Great care should be taken in handling the powder and preparing solutions.] IDENTIFICATION PROCEDURE Standard solution: 1.0 mg/mL of USP Cisplatin RS, 9 mg/mL of sodium chloride, and 10 mg/mL of D-mannitol in water Sample solution: Dissolve the contents of 1 container in water to provide a Cisplatin concentration of 1.0 mg/mL, based on label claim. Solution A: 5.6 g of stannous chloride in 10 mL of hydrochloric acid Stir for 5 min. [NOTEIt is not necessary that all of the solids dissolve.] Solution B: 0.2 g of potassium iodide in 90 mL of water Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Acetone and 1 N nitric acid (9:1) Spray reagent A: Mix Solution A and Solution B together. Disregard any precipitate that is formed. [NOTEStore in the dark. The solution is usable for at least 1 week.] Spray reagent B: 20 mg/mL of potassium iodide in water Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Place the plate in a suitable chromatographic chamber containing a filterpaper lining and equilibrated for 30 min with the Developing solvent system. Develop the plate for a distance of about 8 cm from the origin. Remove the plate, and allow it to air-dry. Complete the drying by heating in a forced-air oven at about 100 for 1 min. Spray the plate with Spray reagent A, heat it in an oven at about 100 for 5 min, cool, and spray with Spray reagent B to bring out the full color of the spots. Acceptance criteria: The principal spot from the Sample solution corresponds in appearance and RF value to that from the Standard solution. ASSAY PROCEDURE Mobile phase: Ethyl acetate, methanol, dimethylformamide, and degassed water (25:16:5:5) Standard solution: 1 mg/mL of USP Cisplatin RS in dimethylformamide [NOTEUse within 1 h.] Sample solution: Quantitatively dissolve the Cisplatin in one container by sonicating for 5 min with dimethylformamide to yield 1.0 mg/mL of Cisplatin. Filter, and discard the first mL of the filtrate.
USP 32
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 310 nm Column: 4.0-mm 30-cm; packing L8 Flow rate: 2.0 mL/min Injection size: 40 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution. Calculate the quantity as a percentage of the label claim of Cl2H6N2Pt in the container: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cisplatin RS in the Standard solution (mg/mL) = nominal concentration of cisplatin in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements W Mr1
Official Monographs / Cisplatin 85

= labeled amount of Cisplatin per container (mg) = molecular weight of trichloroammineplatinate, 318.48 = molecular weight of potassium Mr2 trichloroammineplatinate, 357.58 F = unit conversion factor; 0.001 mg/g Acceptance criteria: NMT 1.0% PROCEDURE: LIMIT OF TRANSPLATIN Mobile phase: 0.18 M monobasic potassium phosphate solution Adjust with phosphoric acid to a pH of 3.2. Standard stock solution: 0.05 mg/mL of USP Transplatin RS in saline TS [NOTEDissolve by stirring by mechanical means for 30 min.] Standard solution A: Add 5 mL of Standard stock solution to 12 mg of USP Cisplatin RS. Dilute with saline TS to 25 mL, and stir by mechanical means for 30 min to dissolve. Standard solution B: Pipet 10 mL of Standard solution A into a 50-mL volumetric flask. Add 5.0 mL of 1 in 200 solution of thiourea (prepared fresh daily) and 5.0 mL of 1 N hydrochloric acid, and dilute with saline TS to volume. Place 10 mL of this solution in a suitable serum vial, seal with a polytef-lined closure, and heat in a heating block at 60 0.5 for 60 min. Remove, and cool to room temperature. Sample solution A: Dissolve the contents of one container with water to yield a nominal 0.5 mg/mL solution of Cisplatin. Sample solution B: Pipet 10 mL of Sample solution A into a 50-mL volumetric flask and proceed as directed for Standard solution B, beginning with add 5.0 mL of a 1 in 200 solution of thiourea. System suitability solution: 0.05 mg/mL of USP Cisplatin RS in saline TS Stir by mechanical means for 30 min to dissolve. Pipet 10 mL of this solution and 10 mL of Standard stock solution into a 50-mL volumetric flask, and proceed as directed for Standard solution B beginning with add 5.0 mL of a 1 in 200 solution of thiourea. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L9 Column temperature: 45 Flow rate: 2.0 mL/min Injection size: 20 L [NOTECondition the column by pumping Mobile phase at a flow rate of 2.0 mL/min for 30 min, then at 0.5 mL/min for 30 min, and then again at 2.0 mL/min for 30 min] System suitability Sample: System suitability solution and Standard solution B [NOTEThe retention time of the derivatized transplatin is 5.09.0 min; or, if it is not, modify the Mobile phase as necessary, and recondition the column. The relative retention times for cisplatin and transplatin are 1.0 and 1.3, respectively.] Suitability requirements Resolution: NLT 1.7, System suitability solution Column efficiency: NLT 2500 theoretical plates, Standard solution B Relative standard deviation: NMT 4.0%, Standard solution B Analysis Samples: Standard solution B and Sample solution B Calculate the percentage of transplatin in the portion of cisplatin taken: Result = (rU/rS) (CS/CU) F 100
IMPURITIES Organic Impurities PROCEDURE: LIMIT OF TRICHLOROAMMINEPLATINATE Mobile phase: 0.4 mg/mL of ammonium sulfate in water The pH of this solution is 5.9 0.1. Make adjustments to the ionic strength of the Mobile phase, if necessary, to meet the System suitability requirements. [NOTEUse low-actinic glassware to prepare the Standard solution and the Sample solution.] Standard solution: 6 g/mL of USP Potassium Trichloroammineplatinate RS in saline TS [NOTEUse within 4 h.] Sample solution: Quantitatively dissolve with water the contents of one container to yield a 0.5 mg/mL solution of Cisplatin. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 209 nm Column: 4.6-mm 25-cm; packing L14 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for cisplatin (in the void volume) and trichloroammineplatinate are 1.0 and 5.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the saline TS and the trichloroammineplatinate peaks Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of trichloroammineplatinate in the portion of cisplatin taken: Result = (rU/rS) (CS V/W) (Mr1/Mr2) F 100 rU rS CS V = = = = peak area from the Sample solution peak area from the Standard solution concentration of Standard solution (g/mL) volume of the constituted container contents (mL)
86
Cisplatin / Official Monographs

= peak area from the Sample solution B = peak area from the Standard solution B = concentration of USP Transplatin RS in the Standard solution B (g/mL) = nominal concentration of cisplatin in the Sample CU solution B (mg/mL) F = unit conversion factor, 0.001 mg/g Acceptance criteria: NMT 2.0% rU rS CS
USP 32
DEFINITION Citalopram Hydrobromide contains NLT 98.0% and NMT 102.0% of C20H21FN2O HBr, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Bromide 191: 10 mg/mL ASSAY PROCEDURE Solution A: 1 mg/mL of sodium acetate in water and triethylamine (497:3). Adjust with acetic acid to a pH of 4.6. Mobile phase: Acetonitrile and Solution A (1:4) The apparent pH is 5.0 0.1. Diluent: Methanol and water (1:1) Standard solution: 0.625 mg/mL of USP Citalopram Hydrobromide RS in Diluent Sample solution: 0.625 mg/mL of Citalopram Hydrobromide in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 239 nm Column: 150-mm 4.6-cm; 5 m packing L7 Temperature: 50 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3000 theoretical plates Tailing factor: NMT 3.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Record the chromatograms for about 30 min. Calculate the percentage of C20H21FN2O HBr, in the portion of Citalopram Hydrobromide taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution rU = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% The sample is moistened with 2 mL of nitric acid and 5 drops of sulfuric acid. HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities [NOTEOn the basis of the synthetic route used, perform either Procedure 1 or Procedure 2. However, if the chloro and bromo analogs are potential related compounds in the synthetic route used, Test-2 is recommended] PROCEDURE 1 Solution A, Mobile phase, Diluent, and Chromatographic system: Proceed as directed in the Assay. Standard stock solution: Use the Standard solution, prepared as directed in the Assay.
SPECIFIC TESTS PH 791: 3.56.2 in the solution constituted as directed in the labeling, using Sterile Water for Injection. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 2.0 USP Endotoxin Units/mg of cisplatin. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. WATER DETERMINATION, Method I 921 Sample: Container contents Analysis: Use anhydrous formamide as the extraction solvent, and use the following procedure. Introduce 50 mL of anhydrous formamide into the titration vessel, and titrate with the Reagent to the electrometric endpoint. Use the formamide thus dried to rinse a suitable glass syringe equipped with a 22-gauge needle, about 8 cm long. Add the rinse back to the titration vessel, and again titrate the vessel contents, if necessary. Via the syringe, withdraw 5 mL of the formamide thus titrated, and, through the closure of the container, expel the contents into the container. Shake the container to obtain a solution. With the same syringe, withdraw all of the contents of the container, and transfer to the titration vessel. Titrate to the endpoint, adjusting the feeding speed control to the lowest setting, to avoid overtitration. Acceptance criteria: NMT 2.0% CONSTITUTED SOLUTIONS: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. OTHER REQUIREMENTS: It meets the requirements for Injections 1, Labels and Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1, Containers for Sterile Solids. Protect from light. USP REFERENCE STANDARDS 11 USP Cisplatin RS USP Endotoxin RS USP Potassium Trichloroammineplatinate RS USP Transplatin RS
Citalopram Hydrobromide
(Comment on this Monograph)id=m17940=Citalopram Hydrobromide=Chl-Cy-Monos.pdf)
405.30 C20H21FN2O HBr 5-Isobenzofurancarbonitrile, 1-[3-(dimethylamino)propyl]-1-(4fluorophenyl)-1,3-dihydro-, monohydrobromide; 1-[3-(Dimethylamino)propyl]-1-(p-fluorophenyl)-5phthalancarbonitrile monohydrobromide [59729-32-7].
USP 32
Standard solution: 0.625 g/mL of citalopram hydrobromide from Standard stock solution in Mobile phase. System suitability solution: 0.001 mg/mL of USP Citalopram Hydrobromide RS and USP Citalopram Related Compound D RS in Diluent Sensitivity solution: Standard solution and Diluent (1:9) Sample solution: Use the Sample solution, prepared as directed under Assay. System suitability Sample: System suitability solution and Sensitivity solution [NOTEFor the purpose of identification, the approximate relative retention times are 0.90 for citalopram related compound D and 1.0 for citalopram hydrobromide.] Suitability requirements Resolution: Between citalopram related compound D and citalopram is NLT 1.8., System suitability solution Tailing factor: 0.81.5 for the citalopram hydrobromide peak, System suitability solution Relative standard deviation: NMT 5.0%, based on the citalopram peak, System suitability solution Signal-to-noise ratio: NLT, Sensitivity solution Analysis Samples: Standard solution and Sample solution Record the chromatograms for about 40 min. Calculate the percentage of related compounds in the portion of Citalopram Hydrobromide taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100/F = peak response for each impurity from the Sample solution rS = peak response for the citalopram peak from the Standard solution CS = concentration of Citalopram Hydrobromide in the Standard solution (mg/mL) = concentration of Citalopram Hydrobromide in CU the Sample solution (mg/mL) = molecular weight of citalopram, 324.39 Mr1 = molecular weight of citalopram hydrobromide, Mr2 405.30 F = relative response factor for each impurity relative to citalopram (free base) Acceptance criteria Individual impurities (See Impurity Table 1.) Total impurities: NMT 0.5%
Impurity Table 1 Impurity Name Relative Retention Time Relative Response Factor 0.34 Acceptance Criteria NMT (%) 0.1
Official Monographs / Citalopram 87

Impurity Table 1 (continued) Impurity Name Citalopram related compound D Citalopram hydrobromide Citalopram related compound E Individual unknown impurity Relative Retention Time 0.90 1.0 1.29 Relative Response Factor 1.04 1.0 0.91 1.0 Acceptance Criteria NMT (%) 0.1 0.1 0.1
PROCEDURE 2 Buffer: 2.7 mg/mL of monobasic potassium phosphate in water Add 1 mL/L of N, N-dimethyloctylamine, stir, and adjust with phosphoric acid to a pH of 3.0. Diluent: Acetonitrile and Buffer (3:7) Solution A: Methanol, tetrahydrofuran and Buffer (24:6:70) Solution B: Acetonitrile and Buffer (4:1) Mobile phase: See the gradient table below.
rU
1-(3-Dimethylamino0.13 propyl)-1-(4fluorophenyl)-5-(4dimethylaminobutyryl)-1,3dihydrobenzofuran Citalopram related compound A 4-[4Dimethylamino-1(4-fluorophenyl)-1hydroxy-1butyl]-3hydroxymethyl benzonitrile Citalopram related compound B Citalopram related compound C 0.18 0.26
0.77 0.99
0.1 0.1
0.40 0.67
0.98 0.69
0.1 0.1
Standard solution: 1.5 g/mL each of USP Citalopram Hydrobromide RS, USP Citalopram Related Compound A RS, USP Citalopram Related Compound C RS, USP Citalopram Related Compound D RS, USP Citalopram Related Compound G RS, and USP Citalopram Related Compound H RS in Diluent Sample solution: 1.5 mg/mL of Citalopram Hydrobromide in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 224 nm Column: 4.6-mm 25-cm; 3-m packing L1 Temperature: 40 Flow rate: 0.8 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEFor the purpose of identification, the approximate relative retention times of citalopram related compounds are provided in Impurity Table 2.] Suitability requirements Resolution: Between citalopram and citalopram related compound D is NLT 2.0 and between citalopram related compound G and citalopram related compound H is NLT 4.0. Relative standard deviation: NMT 2.0% for the citalopram peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of each citalopram related compound in the portion of Citalopram Hydrobromide taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100
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= peak area of each impurity from the Sample solution = peak area of each corresponding impurity from rS the Standard solution = concentration of each citalopram related CS compound in the Standard solution (mg/mL) = concentration of Citalopram Hydrobromide in CU the Sample solution (mg/mL) = molecular weight of citalopram, 324.39 Mr1 = molecular weight of citalopram hydrobromide, Mr1 405.30 Acceptance criteria Individual impurities: See Impurity Table 2. Total specified and unspecified impurities: NMT 0.5%
Impurity Table 2 Impurity Name Relative Retention Time 0.40 0.88 1.0 1.09 2.20 2.30 Relative Response Factor Acceptance Criteria NMT (%) 0.10 0.10 0.10 0.10 0.10 0.10
USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Extract finely ground Tablet powder containing 200 mg of citalopram with 30 mL of water, and filter. Add 1 mL of 1 N sodium hydroxide to the filtrate, and extract with 50 mL of cyclohexane by shaking for 10 min. Pass the cyclohexane layer through a silicone-treated filter paper into a beaker. Reduce the filtrate down to 3 mL, using gentle heat as necessary. Transfer the hot solution to a small centrifuge tube. Induce crystallization while cooling by scratching the side of the test tube with a spatula. Centrifuge the mixture, and decant off the cyclohexane. Dry the residue under vacuum in a desiccator. Mix approximately 2 mg of the residue with approximately 300 mg of potassium bromide, and record the IR spectrum. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 1.42 mg/mL of anhydrous dibasic sodium phosphate in water Diluent: Methanol and Solution A(4:1) Mobile phase: 0.77 mg/mL of dodecyltrimethylammonium bromide in Diluent Internal standard solution: 0.25 mg/mL of USP Citalopram Related Compound F RS in Diluent Standard stock solution: 1.25 mg/mL of USP Citalopram Hydrobromide RS in Diluent Standard solution: Pipet 5.0 mL of the Standard stock solution and 5.0 mL of Internal standard solution into a 50mL volumetric flask. Dilute with Diluent to volume Sample solution: Transfer 10 Tablets to a 200-mL volumetric flask, add 25 mL of Solution A, and shake by mechanical means until disintegrated. Add 100 mL of methanol, and sonicate for about 5 min. Allow to cool to room temperature, then dilute with Diluent to volume. Allow to stand until the residue settles before taking an aliquot for dilution. Transfer a volume of the clear supernatant to a 50mL volumetric flask to obtain a final nominal concentration between 0.090 and 0.10 mg/mL of citalopram. Add 5.0 mL of Internal standard solution, and dilute with Diluent to volume. Pass a portion through a filter (PTFE) having a 0.45m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5 m packing L1 Column temperature: 45 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTERelative retention times are about 1.36 for citalopram related compound F and 1.0 for citalopram.] Suitability requirements Resolution: Between citalopram and citalopram related compound F is NLT 1.5. Column efficiency: NLT 2000 theoretical plates, calculated from the citalopram peak Relative standard deviation: NMT 1.5% for the citalopram peak
rU
Citalopram related compound A Citalopram related compound C Citalopram Citalopram related compound D Citalopram related compound G Citalopram related compound H Individual unspecified impurity
SPECIFIC TESTS SPECIFIC ROTATION 781S: 0.2 to +0.2, at 20 Sample solution: 25 mg/mL, in methanol PH 791: 5.56.5, in 5 mg/mL solution WATER DETERMINATION, Method I 921: NMT 0.5%, with 250 mg COMPLETENESS OF SOLUTION: The absorbance at 410 nm of a 2.5% w/v solution, in 96% alcohol, against a sample solvent in a 1-cm quartz cell is NMT 0.040. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store at controlled room temperature. LABELING: If a test for Organic Impurities other than Procedure 1 is used, then the labeling states with which Organic Impurities test the article complies. USP REFERENCE STANDARDS 11 USP Citalopram Hydrobromide RS USP Citalopram Related Compound A RS USP Citalopram Related Compound C RS USP Citalopram Related Compound D RS USP Citalopram Related Compound G RS USP Citalopram Related Compound H RS
Citalopram Tablets
(Comment on this Monograph)id=m17948=Citalopram Tablets=Chl-Cy-Monos.pdf) DEFINITION Citalopram Tablets contain an amount of Citalopram Hydrobromide equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of citalopram free base (C20H21FN2O).
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of citalopram of label claim in the portion of the Tablets taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 = ratio of the peak response of citalopram to the internal standard from the Sample solution = ratio of the peak response of citalopram to the RS internal standard from the Standard solution CS = concentration of the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of citalopram, 324.39 Mr1 = molecular weight of citalopram hydrobromide, Mr2 405.30 Acceptance criteria: 90.0%110.0% of the labeled amount of C20 H21 FN2 O RU PERFORMANCE TESTS DISSOLUTION 711 Buffer solution: pH 1.5 buffer(prepared by transferring 118 mL of 1 N hydrochloric acid and 82 mL of 1 N sodium hydroxide to a 1000-mL volumetric flask, diluting with water to volume, and adjusting with 1 N sodium hydroxide to a pH of 1.5 Medium: Buffer solution, 800 mL, deaerated Apparatus 1: 100 rpm Time: 30 min Detector: UV 239 nm Sample solutions: Sample per Dissolution 711. Pass through a 0.45-m PVDF filter, dilute with Medium as needed. Standard solution: 12 g/mL of USP Citalopram Hydrobromide RS in Medium Procedure: Calculate the amount of citalopram hydrobromide dissolved, as a percentage:
Official Monographs / Citalopram 89

solution through a membrane filter (PVDF) having a 0.45m or finer porosity, and use the filtrate. Standard solution: Use the Standard solution, prepared as directed in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C20H21FN2O in the portion of sample taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 Mr2 = ratio of the peak responses of citalopram to the internal standard from the Sample solution = ratio of the peak responses of citalopram to the internal standard from the Standard solution = concentration of the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) = molecular weight of citalopram, 324.39 = molecular weight of citalopram hydrobromide, 405.30
absorbance from the Sample solution absorbance from the Standard solution concentration of the Standard solution (mg/mL) molecular weight of citalopram, 324.39 molecular weight of citalopram hydrobromide, 405.30 D = dilution factor of the Sample solution 800 = volume of Medium (mL) L = Tablet label claim of citalopram (mg) Tolerances: NLT 80% (Q) of the labeled amount of citalopram hydrobromide (C20 H21 FN2 O HBr) is dissolved in 30 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Procedure for content uniformity Solution A, Diluent, Internal standard solution, Mobile phase, Chromatographic system and System suitability: Proceed as directed in the Assay. Sample solution: Transfer 1 Tablet to a 100-mL volumetric flask, add 10 mL of Buffer, and shake by mechanical means until disintegrated. Add 40 mL of methanol, and sonicate for about 5 min. Allow to cool to room temperature. Add sufficient volume of Internal standard solution, and dilute to prepare 0.1 mg/mL of citalopram and 0.025 mg/mL of internal standard with Diluent. Pass a portion of this AU AS CS MR1 MR2
= = = = =
IMPURITIES Organic Impurities PROCEDURE Solution A: 3.15 mg/mL of potassium dihydrogen phosphate and 3.60 mg/mL of disodium hydrogen phosphate (Na2 HPO4 12H2 O) in water Mobile phase: Methanol, acetonitrile, and Solution A (38:7:55). Adjust with phosphoric acid to a pH of 6.5. Standard stock solution: 0.5 mg/mL of USP Citalopram Hydrobromide RS in Mobile phase Standard solution: 0.625 g/mL of citalopram hydrobromide from Standard stock solution in Mobile phase Sensitivity solution: 0.05 g/mL of citalopram hydrobromide from Standard solution diluted with Mobile phase Related compounds stock solutions: Separately prepare 0.1 mg/mL each of USP Citalopram Related Compound A RS, USP Citalopram Related Compound B RS, USP Citalopram Related Compound C RS, and USP Citalopram Related Compound E RS in Mobile phase. Peak identification solution: 0.001 mg/mL each of USP Citalopram Related Compound A RS, USP Citalopram Related Compound B RS, USP Citalopram Related Compound C RS, and USP Citalopram Related Compound E RS, using the Standard stock solution as the Diluent System suitability solution: Dilute 0.5 mL of Citalopram related compound C stock solution and 25.0 mL of Standard stock solution with Mobile phase to 50 mL to obtain a solution containing 0.001 mg/mL of Citalopram related compound C and 0.25 mg/mL of citalopram hydrobromide. Sample solution: Transfer 10 Tablets into a 200-mL volumetric flask, add 25 mL of Solution A, and shake by mechanical means until disintegrated. Add 100 mL of a mixture of methanol and water (1:1), mix, and sonicate for about 5 min. Allow to cool, dilute with a mixture of methanol and water (1:1) to volume, and mix thoroughly. Allow the excipients to settle. Dilute as necessary to obtain a final concentration of 0.5 mg/mL of citalopram. Pass a portion of this solution through a polytetrafluoroethylene (PTFE) membrane filter having a 0.45-m or finer porosity, and use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 239 nm Column: 4.6-mm 15-cm; 5-m packing L1 Column temperature: 45 Flow rate: 0.8 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: NLT 3.5 Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 1.5 The citalopram peak shows no shoulders or excessive tailing. Relative standard deviation: NMT 5% Sample: Sensitivity solution Suitability requirements Signal-to-noise ratio: At least 3 Sample: System suitability solution Suitability requirements Resolution: Between citalopram related compound C and citalopram is NLT 3. Sample: Peak identification solution [NOTEFor identification purposes, approximate relative retention times are given in Impurity Table 1.] Suitability requirements Resolution: The four related compound peaks are baseline resolved from each other and the citalopram peak. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in each Tablet taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) (1/F) 100 = peak response for each citalopram related compound from the Sample solution rS = peak response of the corresponding peak in the Standard solution CS = concentration of Citalopram Hydrobromide in the Standard solution (mg/mL) CU = nominal concentration of citalopram hydrobromide in the Sample solution (mg/mL) = molecular weight of citalopram, 324.39 Mr1 = molecular weight of citalopram hydrobromide, Mr2 405.30 F = relative response factor for each impurity relative to citalopram (free base) Acceptance criteria Individual impurities (See Impurity Table 1.) Total impurities: NMT 0.8% rU
Impurity Table 1 Impurity Name Relative Retention Time 0.43 0.60 0.83 1.32 Relative Response Factor 0.77 0.98 0.69 0.91 1.0 Acceptance Criteria NMT (%) 0.2 0.25 0.25 0.1 0.2
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Citalopram Hydrobromide RS USP Citalopram Related Compound A RS USP Citalopram Related Compound B RS USP Citalopram Related Compound C RS USP Citalopram Related Compound E RS USP Citalopram Related Compound F RS
Anhydrous Citric Acid

(Comment on this Monograph)id=m17971=Anhydrous Citric Acid=Chl-Cy-Monos.pdf)
C6H8O7 1,2,3-Propanetricarboxylic acid, 2-hydroxy-; Citric acid [77-92-9].
192.13
DEFINITION Anhydrous Citric Acid contains NLT 99.5% and NMT 100.5% of C6H8O7. IDENTIFICATION INFRARED ABSORPTION 197K Dry the substance to be examined at 105 for 2 h. ASSAY PROCEDURE Sample solution: 0.550 g of Anhydrous Citric Acid in 50.0 mL to a tared flask. Add 0.5 mL of phenolphthalein TS. Analysis: Titrate with 1 N sodium hydroxide VS. Each mL of 1 N sodium hydroxide is equivalent to 64.03 mg of C6H8O7. Acceptance criteria: 99.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1%, determined on 1.0 g SULFATE Standard sulfate solution A: 1.81 mg/mL of potassium sulfate in 30% alcohol. Immediately before use, transfer 10.0 mL of this solution to a 1000-mL volumetric flask and dilute with 30% alcohol to volume. This solution contains 10 g/mL of sulfate. Standard sulfate solution B: 1.81 mg/mL of potassium sulfate in water. Immediately before use, transfer 10.0 mL of this solution to a 1000-mL volumetric flask and dilute to volume. This solution contains 10 g/mL of sulfate. Sample stock solution: 66.7 mg/mL of citric acid Sample solution: To 4.5 mL of Standard sulfate solution A, add 3 mL of a barium chloride solution (1 in 4), shake, and allow to stand for 1 min. To 2.5 mL of the resulting suspension, add 15 mL of the Citric acid stock solution and 0.5 mL of 5 N acetic acid. Standard solution: Prepare as directed for the Sample solution, except use 15 mL of Standard sulfate solution B instead of the Citric acid stock solution.
Citalopram related compound A Citalopram related compound B Citalopram related compound C Citalopram related compound E Any other individual unidentified impurity
USP 32
Analysis Samples: Standard solution and Sample solution Acceptance criteria: Any turbidity produced in the Sample solution after 5 min standing is NMT that produced in the Standard solution (0.015%). HEAVY METALS 231: NMT 10 ppm LIMIT OF ALUMINUM (where it is labeled as intended for use in dialysis) Standard stock solution: To 352 mg of aluminum potassium sulfate in a 100-mL volumetric flask, dissolve with swirling in a few mL of water, add 10 mL of diluted sulfuric acid, and dilute to volume. Immediately before use, mix 1 mL of the resulting solution and dilute to 100 mL. pH 6.0 Acetate buffer: Dissolve 50 g of ammonium acetate with 150 mL, adjust with glacial acetic acid to a pH of 6.0, and dilute to 250 mL. Sample solution: Dissolve 20.0 g of Anhydrous Citric Acid in 100 mL of water, and add 10 mL of pH 6.0 Acetate buffer. Extract this solution with successive portions of 20, 20, and 10 mL of a 0.5% solution of 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50-mL volumetric flask. Dilute the combined extracts with chloroform to volume. Standard solution: Prepare a mixture of 2.0 mL of Standard stock solution, 10 mL of pH 6.0 Acetate buffer, and 98 mL of water. Extract this mixture as described for the Sample solution, dilute the combined extracts with chloroform to volume. Blank solution: Prepare a mixture of 10 mL of pH 6.0 Acetate buffer and 100 mL of water. Extract this mixture as described for the Sample solution, dilute the combined extracts with chloroform to volume. Fluorometric conditions Excitation wavelength: 392 nm Emission wavelength: 518 nm Analysis Samples: Sample solution and Standard solution Acceptance criteria: The fluorescence of the Sample solution does not exceed that of the Standard solution (0.2 ppm). Organic Impurities LIMIT OF OXALIC ACID Sample stock solution: Dissolve 800 mg of Anhydrous Citric Acid in 4 mL. Sample solution: Add 3 mL of hydrochloric acid and 1 g of granular zinc, boil for 1 min, and allow to stand for 2 min. Transfer the supernatant to a test tube containing 0.25 mL of a phenylhydrazine hydrochloride solution (1 in 100), and heat to boiling. Cool rapidly, transfer to a graduated cylinder, and add an equal volume of hydrochloric acid and 0.25 mL of a potassium ferricyanide solution (1 in 20). Shake, and allow to stand for 30 min. Standard solution: Prepare as directed for the Sample solution, except use 4 mL of 0.10 mg/mL oxalic acid solution, equivalent to 0.0714 mg/mL of anhydrous oxalic acid, instead of the Citric acid solution. Analysis Samples: Standard solution and Sample solution Acceptance criteria: Any pink color produced in the Sample solution is not more intense than that produced in the Standard solution (0.036%). SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1.0% BACTERIAL ENDOTOXINS 85: The level of bacterial endotoxins is such that the requirement in the relevant dosage form monograph(s) in which Anhydrous Citric Acid is used can be met. Where the label states that Anhydrous Citric Acid must be subjected to further processing during the preparation of injectable dosage forms, the level of bacterial endotoxins is such that the requirement in the
Official Monographs / Citric 91

relevant dosage form monograph(s) in which Anhydrous Citric Acid is used can be met. STERILITY 71: Where the label states that Anhydrous Citric Acid is sterile, it meets the requirements for Sterility 71 in the relevant dosage form monograph(s) in which Anhydrous Citric Acid is used. READILY CARBONIZABLE SUBSTANCES Sample: 1.0 g of powdered Anhydrous Citric Acid Analysis: Transfer the Sample to a 22- 175-mm test tube previously rinsed with 10 mL of sulfuric acid TS and allowed to drain for 10 min. Add 10 mL of sulfuric acid TS, agitate until solution is complete, and immerse in a water bath at 90 1 for 60 0.5 min, keeping the level of the acid below the level of the water during the entire period. Cool the tube in running water, and transfer the acid to a colorcomparison tube. Acceptance criteria: The color of the acid is not darker than that of a similar volume of Matching Fluid K (see Color and Achromicity 631) in a matching tube, the tubes being observed vertically against a white background. CLARITY OF SOLUTION [NOTEThe Sample solution is to be compared to Standard suspension A in diffused daylight 5 min after preparation of Standard suspension A.] Hydrazine sulfate solution: 10 mg/mL of hydrazine sulfate [NOTEAllow to stand for 4 to 6 h before use.] Methenamine solution: Transfer 2.5 g of methenamine and add 25.0 mL. [NOTEUse a glass-stoppered flask to dissolve.] Primary opalescent suspension: Transfer 25.0 mL of Hydrazine sulfate solution to the Methenamine solution. Mix, and allow to stand for 24 h. [NOTEThis suspension is stable for 2 months, provided it is stored in a glass container free from surface defects. The suspension must not adhere to the glass and must be well mixed before use.] Opalescence standard: Dilute 15.0 mL of Primary opalescent suspension to 1000 mL. [NOTEThis suspension should not be used beyond 24 h after preparation.] Standard suspension A: Dilute 5.0 mL of Opalescence standard to 100 mL. Standard suspension B: Dilute 10.0 mL of Opalescence standard to 100 mL. Sample solution: 200 mg/mL of Anhydrous Citric Acid in water Analysis Samples: Standard suspension A, Standard suspension B, and Sample solution Transfer a sufficient portion of the Sample solution to a test tube of colorless, transparent, neutral glass with a flat base and an internal diameter of 15 to 25 mm to obtain a depth of 40 mm. Similarly transfer portions of Standard suspension A, Standard suspension B , and water to separate matching test tubes. Compare the Sample solution, Standard suspension A, Standard suspension B, and water in diffused daylight, viewing vertically against a black background (see Spectrophotometry and LightScattering 851, Visual Comparison). [NOTEThe diffusion of light must be such that Standard suspension A can readily be distinguished from water, and that Standard suspension B can readily be distinguished from Standard suspension A.] Acceptance criteria: The Sample solution shows the same clarity as that of water. COLOR OF SOLUTION Standard stock solution A: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (10 g/L) (2.4:0.6:0:7.0)
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USP 32
ASSAY PROCEDURE Sample solution: 0.550 g of Citric Acid Monohydrate in 50.0 mL to a tared flask. Add 0.5 mL of phenolphthalein TS. Analysis: Titrate with 1 N sodium hydroxide VS. Each mL of 1 N sodium hydroxide is equivalent to 64.03 mg of C6H8O7. Acceptance criteria: 99.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1%, determined on 1.0 g SULFATE Standard sulfate solution A: 1.81 mg/mL of potassium sulfate in 30% alcohol. Immediately before use, transfer 10.0 mL of this solution to a 1000-mL volumetric flask, and dilute with 30% alcohol to volume. This solution contains 10 g/mL of sulfate. Standard sulfate solution B: 1.81 mg/mL of potassium sulfate. Immediately before use, transfer 10.0 mL of this solution to a 1000-mL volumetric flask, and dilute to volume. This solution contains 10 g/mL of sulfate. Sample stock solution: 66.7 mg/mL of Citric Acid Monohydrate Sample solution: To 4.5 mL of Standard sulfate solution A, add 3 mL of a barium chloride solution (1 in 4), shake, and allow to stand for 1 min. To 2.5 mL of the resulting suspension, add 15 mL of the Sample stock solution and 0.5 mL of 5 N acetic acid, and mix. Standard solution: Prepare like the Sample solution, except use 15 mL of Standard sulfate solution B instead of Sample stock solution. Analysis Samples: Standard solution and Sample solution Acceptance criteria: Any turbidity produced in the Sample solution after 5 min standing is NMT that produced in the Standard solution (0.015%). HEAVY METALS 231: NMT 10 ppm LIMIT OF ALUMINUM (where it is labeled as intended for use in dialysis) Standard aluminum solution: To 352 mg of aluminum potassium sulfate in a 100-mL volumetric flask, dissolve with swirling in a few mL of water, add 10 mL diluted sulfuric acid, and dilute with water to volume. Immediately before use, mix 1 mL of the resulting solution, and dilute with water to 100 mL. pH 6.0 acetate buffer: Dissolve 50 g of ammonium acetate with 150 mL, adjust with glacial acetic acid to a pH of 6.0, and dilute to volume to 250 mL. Sample solution: Dissolve 20.0 g of Citric Acid Monohydrate in 100 mL, and add 10 mL of pH 6.0 acetate buffer. Extract this solution with successive portions of 20, 20, and 10 mL of a 0.5% solution of 8-hydroxyquinoline in chloroform, combining the chloroform extracts in a 50-mL volumetric flask. Dilute the combined extracts with chloroform to volume. Standard solution: Prepare a mixture of 2.0 mL of Standard aluminum solution, 10 mL of pH 6.0 acetate buffer, and 98 mL of water. Extract this mixture as described for the Sample solution, and dilute the combined extracts with chloroform to volume. Blank solution: Prepare a mixture of 10 mL of pH 6.0 acetate buffer and 100 mL of water. Extract this mixture as described for the Sample solution, and dilute the combined extracts with chloroform to volume.
Standard stock solution B: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (10 g/L) (2.4:1.0:0.4:6.2) Standard stock solution C: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (10 g/L) (9.6:0.2:0.2:0) Standard solution A: Transfer 2.5 mL of Standard stock solution A and dilute with dilute hydrochloric acid to 100 mL (10 g/L). Standard solution B: Transfer 2.5 mL of Standard stock solution B and dilute with dilute hydrochloric acid to 100 mL (10 g/L). Standard solution C: Transfer 0.75 mL of Standard stock solution C and dilute with dilute hydrochloric acid to 100 mL (10 g/L). [NOTEPrepare the Standard solutions immediately before use.] Standard solution D: Water Sample solution: Use the Sample solution prepared as directed in the test for Clarity of solution. Analysis Samples: Standard solution A, Standard solution B, Standard solution C, Standard solution D, and Sample solution Transfer portions of each of the Samples to individual test tubes of colorless, transparent, neutral glass with a flat base and an internal diameter of 15 to 25 mm to obtain a depth of 40 mm. Compare Samples against diffuse daylight, viewing vertically against a white background (see Spectrophotometry and Light-Scattering 851, Visual Comparison). Acceptance criteria: The Sample solution is not more intensely colored than Standard solution A, B, C, or D. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. No storage requirements are specified. LABELING: Where it is intended for use in dialysis solutions, it is so labeled. Where Anhydrous Citric Acid must be subjected to further processing during the preparation of injectable dosage forms to ensure acceptable levels of bacterial endotoxins, it is so labeled. Where Anhydrous Citric Acid is sterile, it is so labeled. USP REFERENCE STANDARDS 11 USP Citric Acid RS USP Endotoxin RS
Citric Acid Monohydrate

(Comment on this Monograph)id=m17972=Citric Acid Monohydrate=Chl-Cy-Monos.pdf)
210.14 C6H8O7 H2O 1,2,3-Propanetricarboxylic acid, 2-hydroxy-, monohydrate [5949-29-1]. DEFINITION Citric Acid Monohydrate contains one molecule of water of hydration. It contains NLT 99.5% and NMT 100.5% of C6H8O7, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197K: examined at 105 for 2 h. Dry the substance to be
USP 32
Fluorometric conditions Excitation wavelength: 392 nm Emission wavelength: 518 nm Analysis Samples: Standard solution and Sample solution Acceptance criteria: The fluorescence of the Sample solution does not exceed that of the Standard solution (0.2 ppm). Organic Impurities PROCEDURE: LIMIT OF OXALIC ACID Standard stock solution: Dissolve 800 mg of Citric Acid Monohydrate in 4 mL. Sample solution: Add 3 mL of hydrochloric acid and 1 g of granular zinc, boil for 1 min, and allow to stand for 2 min. Transfer the supernatant to a test tube containing 0.25 mL of a phenylhydrazine hydrochloride solution (1 in 100), and heat to boiling. Cool rapidly, transfer to a graduated cylinder, and add an equal volume of hydrochloric acid and 0.25 mL of a potassium ferricyanide solution (1 in 20). Shake and allow to stand for 30 min. Standard solution: Prepare like the Sample solution, except use 4 mL of 0.10 mg/mL oxalic acid solution, equivalent to 0.0714 mg/mL of anhydrous oxalic acid, instead of the Sample stock solution. Analysis Samples: Standard solution and Sample solution Acceptance criteria: Any pink color produced in the Sample solution is not more intense than that produced in the Control solution (0.036%). SPECIFIC TESTS WATER DETERMINATION, Method I 921: 7.5%9.0% BACTERIAL ENDOTOXINS TEST 85: The level of bacterial endotoxins is such that the requirement in the relevant dosage form monograph(s) in which Citric Acid Monohydrate is used can be met. Where the label states that Citric Acid Monohydrate must be subjected to further processing during the preparation of injectable dosage forms, the level of bacterial endotoxins is such that the requirement in the relevant dosage form monograph(s) in which Citric Acid Monohydrate is used can be met. STERILITY TESTS 71: Where the label states that Citric Acid Monohydrate is sterile, it meets the requirements for Sterility Tests 71 in the relevant dosage form monograph(s) in which Citric Acid Monohydrate is used. READILY CARBONIZABLE SUBSTANCES Sample: 1.0 g powdered Citric Acid Monohydrate Analysis: Transfer Sample to a 22- 175-mm test tube previously rinsed with 10 mL of sulfuric acid TS and allowed to drain for 10 min. Add 10 mL of sulfuric acid TS, agitate until solution is complete, and immerse in a water bath at 90 1 for 60 0.5 min, keeping the level of the acid below the level of the water during the entire period. Cool the tube in running water, and transfer the acid to a colorcomparison tube. Acceptance criteria: The color of the acid is not darker than that of a similar volume of Matching Fluid K (see Color and Achromicity 631) in a matching tube, the tubes being observed vertically against a white background. CLARITY OF SOLUTIONS [NOTEThe Sample solution is to be compared to Reference suspension A in diffused daylight 5 min after preparation of Reference suspension A.] Hydrazine sulfate solution: 10 mg/mL of hydrazine sulfate [NOTEAllow to stand for 46 h before use.] Methenamine solution: Transfer 2.5 g of Methenamine and add 25.0 mL. [NOTEUse a glass-stoppered flask to dissolve.] Primary opalescent suspension: Transfer 25.0 mL Hydrazine sulfate solution to the Methenamine solution. Allow to stand for 24 h. [NOTEThis suspension is stable for 2 months, provided it is stored in a glass container free from
Official Monographs / Citric 93

surface defects. The suspension must not adhere to the glass and must be well mixed before use.] Opalescence standard: Dilute 15.0 mL Primary opalescent suspension to 1000 mL. [NOTEThis suspension should not be used beyond 24 h after preparation.] Standard suspension A: Dilute 5.0 mL Opalescence standard to 100 mL. Standard suspension B: Dilute 10.0 mL Opalescence standard to 100 mL. Sample solution: 200 mg/mL of Citric Acid Monohydrate Analysis Samples: Standard suspension A, Standard suspension B, water and Sample solution Transfer a sufficient portion of the Samples to individual test tube of colorless, transparent, neutral glass with a flat base and an internal diameter of 1525 mm to obtain a depth of 40 mm. Compare the Samples in diffused daylight, viewing vertically against a black background (see Spectrophotometry and Light-Scattering 851, Visual Comparison). [NOTEThe diffusion of light must be such that Standard suspension A can readily be distinguished from water, and that Standard suspension B can readily be distinguished from Standard suspension A.] Acceptance criteria: The Sample solution shows the same clarity as that of water. COLOR OF SOLUTION Standard stock solution A: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (10 g/L) (2.4:0.6:0:7.0) Standard stock solution B: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (10 g/L) (2.4:1.0:0.4:6.2) Standard stock solution C: Ferric chloride CS, cobaltous chloride CS, cupric sulfate CS, and dilute hydrochloric acid (10 g/L) (9.6:0.2:0.2:0) Standard solution A: Transfer 2.5 mL of Standard stock solution A, and dilute with dilute hydrochloric acid (10 g/L) to 100 mL. Standard solution B: Transfer 2.5 mL of Standard stock solution B, and dilute with dilute hydrochloric acid (10 g/L) to 100 mL. Standard solution C: Transfer 0.75 mL of Standard stock solution C, and dilute with dilute hydrochloric acid (10 g/L) to 100 mL. [NOTEPrepare the Standard solutions immediately before use.] Standard solution D Sample solution: Use the Sample solution prepared as directed in the test for Clarity of solutions. Analysis Samples: Standard solution A, Standard solution B, Standard solution C, and Sample solution Transfer a sufficient portion of the Samples to individual test tube of colorless, transparent, neutral glass with a flat base and an internal diameter of 1525 mm to obtain a depth of 40 mm. Compare the Samples in diffused daylight, viewing vertically against a white background (see Spectrophotometry and Light-Scattering 851, Visual Comparison). Acceptance criteria: The Sample solution is not more intensely colored than Standard solution A, Standard solution B, Standard solution C, or Standard solution D. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. No storage requirements specified. LABELING: Where it is intended for use in dialysis solutions, it is so labeled. Where Citric Acid Monohydrate must be subjected to further processing during the preparation of injectable dosage forms to ensure acceptable levels of bacterial endotoxins, it is so labeled. Where Citric Acid Monohydrate is sterile, it is so labeled.
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Citric / Official Monographs
USP 32
PROCEDURE 2: CITRIC ACID Mobile phase, Standard solution 1, and Chromatographic system: Proceed as directed under Assay for Citric Acid/Citrate and Phosphate 345. Sample solution: Transfer equivalent to 130 mg of monohydrate citric acid from Irrigation, into a suitable volumetric flask, and proceed as directed under Assay for Citric Acid/Citrate and Phosphate 345, Assay Preparation for Citric Acid/Citrate Assay. Analysis Samples: Standard solution 1 and Sample solution Proceed as directed under 345, Procedure, and calculate the percentage of monohydrate citric acid (C6H8O7 H2O) in the portion of Irrigation taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 citrate peak area from the Sample solution citrate peak area from the Standard solution concentration of Standard solution 1(g/mL) nominal concentration of the Sample solution (unit/mL) Mr1 = molecular weight of citric acid monohydrate, 210.14 Mr2 = molecular weight of citrate, 189.10 Acceptance criteria: 95.0%105.0% of the labeled amount of C6H8O7 H2O (Official January 1, 2009) PROCEDURE 3: MAGNESIUM OXIDE Sample solution: A volume of Irrigation, nominally equivalent to 40 mg of magnesium oxide Analysis: Transfer Sample solution to a beaker containing 130 mL of water heated to 75 5, and add 4 mL of ammonium chloride TS and then 5 mL of ammonium hydroxide. Mix, and add slowly, with stirring, 8 mL of 8hydroxyquinoline TS. After allowing to stand for 30 min at 75, filter through a sintered-glass crucible, previously dried and weighed, and wash the precipitate with 50 mL of a warm mixture of water and 6 N ammonium hydroxide (45:5), followed by 50 mL of cool water. Dry the crucible and contents at 105 for 3 h, cool, and weigh. Determine the equivalent of MgO in the portion of Irrigation taken by multiplying the weight of C18H12MgN2O2 2H2O so obtained by 0.1156 Acceptance criteria: 95.0%105.0% of the labeled amounts of MgO PROCEDURE 4: SODIUM CARBONATE Sodium chloride stock solution: 4.75 mg/mL of sodium chloride, previously dried at 105 for 2 h, in water Internal standard solution: 0.636 mg/mL of lithium chloride in water Standard solution: Prepare a mixture of Internal standard solution and Sodium chloride stock solution (99:1). Sample stock solution: Nominally equivalent to 4.4 mg/mL sodium carbonate from Irrigation diluted with water Sample solution: Internal standard solution and Sample stock solution (99:1) Spectrometric conditions Mode: Flame photometer Analytical wavelengths: 591 nm and 671 nm Analysis: Concomitantly determine the emittances of the Standard solution and the Sample solution, adjusting the instrument to zero emittance with Internal standard solution. Calculate the percentage of the label claim of Na2CO3 in the Irrigation taken: rU rS CS CU Result = (rU,591/rU,671)(rS,671/rS,591) (CS/Cu) (MW1/MW2) 100 = = = =
USP REFERENCE STANDARDS 11 USP Citric Acid RS
Citric Acid, Magnesium Oxide, and Sodium Carbonate Irrigation

(Comment on this Monograph)id=m17980=Citric Acid, Magnesium Oxide, and Sodium Carbonate Irrigation=Chl-CyMonos.pdf) DEFINITION Citric Acid, Magnesium Oxide, and Sodium Carbonate Irrigation is a sterile solution of Citric Acid, Magnesium Oxide, and Sodium Carbonate in Water for Injection. It contains NLT 95.0 %and NMT 105.0 %of the labeled amounts of citric acid (C6H8O7 H2O), magnesium oxide (MgO), and sodium carbonate (Na2CO3). IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Sodium, Magnesium 191 B. To 10 mL of the Irrigation add 1 mL of mercuric sulfate TS, heat to boiling, and add a few drops of potassium permanganate TS: a white precipitate is formed. ASSAY PROCEDURE 1: CITRIC ACID Mobile phase: Sulfuric acid and water (1:499). Pass through a membrane filter, heat to 40, and degas. [NOTEMaintain the temperature of the Mobile phase at 40 throughout the analysis.] System suitability solution: 1 mg/mL of citric acid and 2 mg/mL of boric acid in water Standard solution: 1.2 mg/mL of anhydrous citric acid in water Sample solution: Equivalent to 1.3 mg/mL of citric acid monohydrate from Irrigation, in water Chromatographic system Mode: LC Detector: Refractive index Column: 7.8-mm 30-cm; packing L17 Temperature: 40 Flow rate: 0.6 mL/min Injection size: 50 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: Between citric acid and boric acid is NLT 4.0 (System suitability solution) Relative standard deviation: NMT 1.5% (Standard solution) Analysis Samples: Standard solution and Sample solution Calculate the percentage of the label claim of C6H8O7 H2O in the portion of Irrigation taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 peak response of the Sample solution peak response of the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of Sodium carbonate in the Sample solution (mg/mL) = molecular weight of citric acid monohydrate, Mr1 210.14 = molecular weight of citrate, 192.12 Mr2 Acceptance criteria: 95.0%105.0% of the labeled amount of C6H8O7 H2O (Official until January 1, 2009) rU rS CS CU = = = =
USP 32
= emittance reading from the Sample solution at the wavelength indicated by the subscript, 591 rU,671 = emittance reading from the Sample solution at the wavelength indicated by the subscript, 671 = emittance reading from the Standard solution at rS,671 the wavelength indicated by the subscript, 671 = emittance reading from the Standard solution at rS,591 the wavelength indicated by the subscript, 591 = concentration of sodium chloride in the Standard CS solution (mg/mL) = nominal concentration of sodium carbonate in CU the Sample solution (mg/mL) = molecular weight of sodium carbonate, 105.99 Mr1 = two times the molecular weight of sodium Mr2 chloride, 116.88 Acceptance criteria: 95.0%105.0% of the labeled amounts of Na2CO3 SPECIFIC TESTS PH 791: 3.84.2 BACTERIAL ENDOTOXINS 85: It contains NMT 2.80 Endotoxin Units/mL. OTHER REQUIREMENTS: It meets the requirements under Injections 1, except that the container may be designed to empty rapidly, and may exceed 1000 mL in capacity. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I or Type II glass. USP REFERENCE STANDARDS 11 USP Endotoxin RS USP REFERENCE STANDARDS 11 USP Citric Acid RS USP Endotoxin RS (Official January 1, 2009) rU,591
Official Monographs / Cladribine 95

System suitability solution: 0.02 mg/mL each of USP Cladribine RS and USP Cladribine Related Compound A RS in Diluent Standard solution: 0.5 mg/mL of USP Cladribine RS in Diluent Sample solution: 0.5 mg/mL of Cladribine in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1.0 mL/min Injection size: 10 L System suitability Sample: Standard solution and System suitability solution Suitability requirements Resolution: Between cladribine and cladribine related compound A is NLT 1.5., System suitability solution Tailing factor: NMT 2.0 for the cladribine peak, Standard solution Relative standard deviation: NMT 2.0% , Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10 H12 ClN5 O3 in the portion of Cladribine taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cladribine RS in the Standard solution (mg/mL) = concentration of Cladribine in the Sample CU solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Buffer solution, Diluent, Mobile phase, System suitability solution, Standard solution, Suitability requirements, Chromatographic system, and System suitability: Prepare as directed under Assay. Sample solution: Use the Sample solution, prepared as directed in the Assay. Analysis Samples: Sample solution Calculate the percentage of each impurity in the portion of Cladribine taken: Result = (rU/rT) 100 = peak response of each individual impurity rU = sum of the responses of all the peaks rT Acceptance criteria Individual impurities: (See Impurity Table 1.) Total impurities: NMT 1.0%
Relative Retention Time 0.41 0.47 Relative Response Factor Acceptance Criteria NMT (%) 0.20 0.20
rU rS CS
Cladribine
(Comment on this Monograph)id=m17984=Cladribine=Chl-CyMonos.pdf)
C10 H12 ClN5 O3 Adenosine, 2-chloro-2-deoxy-; 2-Chloro-2-deoxyadenosine [4291-63-8].
285.69
DEFINITION Cladribine contains NLT 98.0 %and NMT 102.0 %of C10 H12 ClN5 O3, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer solution: Dissolve 9.96 g of triethylamine phosphate in 500 mL of water. Add another 500 mL of water and adjust pH to 6.1 with potassium hydroxide. Diluent: Methanol and water (1:9) Mobile phase: Filtered and degassed mixture of methanol and Buffer solution (11:39)
Impurity Name 2,6-Diaminopurine-2deoxyriboside 2-Deoxyadenosine
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Cladribine / Official Monographs

Relative Retention Time 0.60 0.91 Relative Response Factor Acceptance Criteria NMT (%) 0.20 0.20
USP 32
= peak response of the Standard solution = concentration of USP Clarithromycin RS in the Standard solution (mg/mL) = concentration of Clarithromycin in the Sample CU solution (mg/mL) Acceptance criteria: 96.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Sample: 0.5 g Analysis: Moisten the charred residue with 1 mL of sulfuric acid. Acceptance criteria: NMT 0.2% HEAVY METALS 231 Diluent: Dioxane and water (85:15) Sample solution: 50 mg/mL in Diluent Transfer 12 mL of this solution to a color-comparison tube. Blank: Add 10 mL of Diluent and 2 mL of the Sample solution to a color-comparison tube. Standard solution: Prepare using standard lead solution (1 ppm Pb) obtained by diluting standard lead solution (100 ppm Pb) with Diluent. Add 10 mL of this solution (1 ppm Pb) and 2 mL of the Sample solution to a color-comparison tube. To each of the three tubes containing the Sample solution, the Blank, and the Standard solution add 2 mL of pH 3.5 acetate buffer, mix, add 1.2 mL of thioacetamideglycerin base TS, and mix. Compared to the Blank, the Standard solution shows a slight brown color. After 2 min, any brown color in the Sample solution is not more intense than that in the Standard solution. Acceptance criteria: NMT 20 ppm Organic Impurities PROCEDURE Solution A: 4.76 g/L of monobasic potassium phosphate. Adjust with dilute phosphoric acid (l in 10) or potassium hydroxide (45% w/v) to a pH of 4.4. Pass this solution through a C18 filtration kit. Solution B: Acetonitrile Mobile phase: Use variable mixtures of Solution A and Solution B as directed under Table 1.
Table 1 Time (min) 0 32 34 36 42 Solution A (%) 75 40 40 75 75 Solution B (%) 25 60 60 25 25
Impurity Name 2-Chloroadenine 2-Methoxy-2deoxyadenosine (cladribine related compound A) Any individual unspecified impurity
rS CS
0.10
SPECIFIC TESTS SPECIFIC ROTATION 781S: Between 17.0 and 21.0 Sample solution: 10 mg/mL, in dimethylformamide WATER DETERMINATION, Method I 921: NMT 4.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and protect from light. Store between 2 and 8. USP REFERENCE STANDARDS 11 USP Cladribine RS USP Cladribine Related Compound A RS
Clarithromycin
(Comment on this Monograph)id=m17990=Clarithromycin=Chl-Cy-Monos.pdf)
C38H69NO13 Erythromycin, 6-O-methyl-; 6-O-Methylerythromycin [81103-11-9]. DEFINITION Clarithromycin contains NLT 96.0% and NMT 102.0% of C38H69NO13, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197K
747.95
ASSAY PROCEDURE Solution A, Solution B, Mobile phase, Diluent, and Standard solution D: Proceed as directed in the Procedure under Organic Impurities. Standard solution: Use Standard solution A, prepared as directed in the Procedure under Organic Impurities. Sample solution: Use the Sample solution, prepared as directed in the Procedure under Organic Impurities. Chromatographic system: Proceed as directed in the Procedure under Organic Impurities. System suitability: Proceed as directed in the Procedure under Organic Impurities. In addition, the relative standard deviation for replicate injections of the Standard solution is NMT 1.5%. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C38H69NO13 in the portion taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of the Sample solution
Diluent: Acetonitrile and water (1:1) Standard solution A: 1.5 mg/mL of USP Clarithromycin RS in acetonitrile and water (1:1) [NOTEDissolve in acetonitrile and then add water.] Standard solution B: 0.075 mg/mL from Standard solution A in Diluent Standard solution C: 0.0075 mg/mL from Standard solution B in Diluent Standard solution D: 1.5 mg/mL of USP Clarithromycin Identity RS in acetonitrile and water (1:1) [NOTEDissolve in acetonitrile and then add water.] Sample solution: 1.5 mg/mL of Clarithromycin in acetonitrile and water (1:1) [NOTEDissolve in acetonitrile and then add water.] Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 205 nm Column: 4.6-mm 10-cm; packing L1 Column temperature: 40 Flow rate: 1.1 mL/min Injection size: 10 L System suitability Sample: Standard solution B [NOTEsee relative retention times below]
Relative Retention Time about 0.38 about 0.42 about 0.63 about 0.74 about 0.79 about 0.81 about 0.89 about 0.96 about 1.15 about 1.27 about 1.33 about 1.35 about 1.38 about 1.59 about 1.72 about 1.82 about 11 min
Official Monographs / Clarithromycin 97

= 1.0, or the correction factor of 0.27, and 0.15 applied to the responses for peaks at relative retention times in relation to that of clarithromycin of about 1.72, and 1.82, corresponding to related compound G and related compound H, respectively Acceptance criteria Any single related compound: NMT 1.0%, NMT four related compounds exceed the limit of 0.4% Total of all related compounds: NMT 3.5% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S Sample solution: 10 mg/mL, in methylene chloride Acceptance criteria: 94 to 102 (at 20) CRYSTALLINITY 695: Meets the requirements PH 791: 8.010.0 Sample: 1-in-500 suspension in water and methanol (19:1) WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clarithromycin RS USP Clarithromycin Identity RS F
Name Impurity I Impurity A Impurity J Impurity L Impurity B Impurity M Impurity C Impurity D Impurity N Impurity E Impurity F Impurity P Impurity O Impurity K Impurity G Impurity H Clarithromycin
Clarithromycin for Oral Suspension

(Comment on this Monograph)id=m17993=Clarithromycin for Oral Suspension=Chl-Cy-Monos.pdf) DEFINITION Clarithromycin for Oral Suspension is a dry mixture of Clarithromycin, dispersing agents, diluents, preservatives, and flavorings. It contains NLT 90.0% and NMT 115.0% of the labeled amount of clarithromycin (C38H69NO13), the labeled amount being 25 mg or 50 mg/mL when constituted as directed in the labeling. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol and 0.067 M monobasic potassium phosphate (3:2), adjust with phosphoric acid to a pH of 3.5. Pass through a filter having a 0.5-m or finer porosity. Standard stock solution: Equivalent to 2100 g/ml of clarithromycin from USP Clarithromycin RS Standard solution: 415 g/ml of clarithromycin from USP Clarithromycin RS from Standard stock solution diluted with Mobile phase. Sample solution: Constitute Clarithromycin for Oral Suspension as directed in the labeling. Transfer equivalent to 1 to 2 g of clarithromycin from Oral Suspension, with the aid of 330 mL of 0.067 M dibasic potassium phosphate, to a 1000-mL volumetric flask containing 50 mL of 0.067 M dibasic potassium phosphate. Shake by mechanical means for 30 min, and dilute with methanol to volume. Sonicate for about 30 min, and allow to cool. Dilute with methanol to volume, add a magnetic stirring bar, and stir for 60 min. Allow to settle, and transfer volume of the clear supernatant, nominally equivalent to 20 mg of clarithromycin, to a 50-mL
Suitability requirements Tailing factor: NMT 1.7 for the main clarithromycin peak System suitability Sample: Standard solution D Suitability requirements Peak-to-valley ratio (HP/HV): NLT 3.0 from the ratio of Impurity D and clarithromycin, where: HP = height above the baseline of the peak due to Impurity D = height above the baseline of the lowest point of HV the curve separating this peak from the peak due to clarithromycin Analysis Samples: Diluent, Standard solution B, Standard solution C, Standard solution D, and the Sample solution Calculate the percentage of each impurity in the Clarithromycin taken: Result = (rU/rS) (CS/CU) F 100 rU rS CS CU = peak response for any individual impurity observed in the chromatogram from the Sample solution = peak response of the main clarithromycin peak of the Standard solution C = concentration of USP Clarithromycin RS in Standard solution C (mg/mL) = concentration of Clarithromycin in the Sample solution (mg/mL)
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Clarithromycin / Official Monographs
USP 32
Adjust with phosphoric acid to a pH of 4.0, and then pass through a filter having a 0.5-m or finer porosity. Standard stock solution: 625 g/mL of clarithromycin from USP Clarithromycin RS dissolved in methanol. [NOTEshake and sonicate to facilitate dissolution.] Standard solution: 125 g/mL of clarithromycin from Standard stock solution diluted with Mobile phase through a 0.5 m porosity filter. System suitability solution: Prepare 625 g/mL of USP Clarithromycin Related Compound A RS in methanol. Transfer 10 mL of this solution and 10 mL of Standard stock solution to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution: Finely powder a counted number of Tablets, nominally equivalent to 2000 mg of clarithromycin, and with the aid of methanol, transfer the powder to a 500mL volumetric flask, add 350 mL of methanol, and shake by mechanical means for 30 min. Dilute with methanol to volume, and allow any insoluble matter to settle. Transfer 3.0 mL to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Filter through a filter having a 0.5-m or finer porosity. Use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 15-cm; packing L1 Column temperature: 50 Flow rate: 1 mL/min Injection size: 2050 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clarithromycin and Carithromycin Related Compound A are 0.75 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between clarithromycin and Clarithromycin Related Compound A, System suitability solution Column efficiency: NLT 750 theoretical plates from the clarithromycin peak, Standard solution Tailing factor: 0.91.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate percentage of label claim of C38H69NO13 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 peak area response of the Sample solution peak area response of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Buffer solution: Transfer 13.61 g of sodium acetate trihydrate to a 1-L volumetric flask, add water to dissolve, dilute with water to volume. Adjust with 0.1 M acetic acid to a pH of 5.0. rU rS CS CU = = = =
volumetric flask, dilute with Mobile phase to volume, and filter through a filter having a 0.5-m or finer porosity. Use the filtrate as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Guard column: Packing L1 (optional) Column: 4.6-mm 15-cm; packing L1 Column temperature: 50 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Column efficiency, n: NLT 2100 theoretical plates from the clarithromycin peak Tailing factor: 1.01.7 Capacity factor, k: 2.56 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C38H69NO13 in each mL of the constituted Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 peak area response from the Sample solution peak area response from the Standard solution concentration of Standard solution (g/mL) nominal concentration of clarithromycin in the Sample solution (g /mL) Acceptance criteria: 90.0%115.0% rU rS CS CU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 (for powder packaged in single-unit containers): Meets the requirements DELIVERABLE VOLUME 698 (for powder packaged in multipleunit containers): Meets the requirements SPECIFIC TESTS PH 791: 4.05.4, in the suspension constituted as directed in the labeling LOSS ON DRYING 731: Dry 1 g of it in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clarithromycin RS USP Clarithromycin Related Compound A RS = = = =
Clarithromycin Tablets
(Comment on this Monograph)id=m17995=Clarithromycin Tablets=Chl-Cy-Monos.pdf) DEFINITION Clarithromycin Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C38H69NO13. IDENTIFICATION The retention time of the major peak from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol and 0.067 M monobasic potassium phosphate (13:7)
USP 32
Medium: Buffer solution, 900 mL. Apparatus: 2: 50 rpm Time: 30 min Standard solution: Proceed as directed under Assay. Sample solution: Sample per Dissolution 711. Dilute with Mobile phase to yield a nominal concentration of 125 g/ml charithromycin. Analysis Sample: Standard solution and Sample solution Determine the amount of C38H69NO13 dissolved in the Medium, as directed in the Assay. Calculate the percentage of clarithromycin dissolved as follows: Result = (rU/rS) (CS/CU) 100 peak area response of the Sample solution peak area response of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (g /mL) Tolerances: NLT 80% (Q) of the labeled amount of C38H69NO13 is dissolved in 30 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU rS CS CU SPECIFIC TESTS LOSS ON DRYING 731: Dry a portion of powdered Tablets in vacuum at a pressure not exceeding 5 mm of mercury at 110 for 3 h: it loses NMT 6.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clarithromycin RS USP Clarithromycin Related Compound A RS = = = =

Sample solution: Transfer a nominal equivalent of 2000 mg of clarithromycin, from finely powdered Tablets, to a 500mL volumetric flask with the aid of methanol. Add 350 mL of methanol, and shake by mechanical means for 30 min. Dilute with methanol to volume. Sonicate for 30 min. Cool to room temperature, and allow to stand for at least 16 h. Allow any insoluble matter to settle. Transfer 3.0 mL of the supernatant to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Filter through a filter having a 0.5 m or finer porosity. Use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Guard column: Packing L1 (optional) Column: 4.6-mm 15-cm; packing L1 Column temperature: 50 Flow rate: 1 mL/min Injection size: 2050 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for clarithromycin and clarithromycin related compound A are about 0.75 and 1.0, respectively, in System suitability solution.] Suitability requirements Resolution: NLT 2.0 between clarithromycin and clarithromycin related compound A, System suitability solution Column efficiency: NLT 750 theoretical plates from the clarithromycin peak, Standard solution Tailing factor: NLT 0.9 and NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of label claim of C38H69NO13 in the Extended-Release Tablets: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Clarithromycin RS in the Standard solution (g/mL) = nominal concentration of clarithromycin in the CU Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Test 1 Buffer solution: 0.3 M phosphate buffer, pH 6.0 0.5 [NOTE 40.83 mg/mL monophosphate potassium phosphate and 24 mg/ml sodium hydroxide, pH adjusted with either phosphoric acid or 1 N sodium hydroxide.] Medium: Buffer solution, 900 mL Apparatus 2: 75 rpm Times: 30, 45, 60, and 120 min Determine the percentages of the labeled amount of C38H69NO13 dissolved using the following method. Standard solutions: Prepare five solutions of USP Clarithromycin RS dissolved in acetonitrile and diluted in Medium, with known concentrations over the range of about 60600 g/mL. Sample solution: Use portions of the solution under test passed through a 35-m polyethylene filter. Chromatographic system: Proceed as directed in the Assay except the Injection size is 50 L.
Clarithromycin Extended-Release Tablets

(Comment on this Monograph)id=m17997=Clarithromycin Extended-Release Tablets=Chl-Cy-Monos.pdf) DEFINITION Clarithromycin Extended-Release Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of clarithromycin (C38H69NO13). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol and 0.067 M monobasic potassium phosphate (13:7) Adjust with phosphoric acid to a pH of 4.0 . Pass through a filter having a 0.5-m or finer porosity. Standard stock solution: 625 g/mL of clarithromycin from USP Clarithromycin RS in methanol, shaking and sonicating if necessary to effect dissolution. Standard solution: 125 g/mL of clarithromycin from Standard stock solution diluted with Mobile phase, filtered through a 0.5-m or finer porosity filter System suitability stock solution: Prepare 625 g/mL of USP Clarithromycin Related Compound A RS in methanol. System suitability solution: 125 g/mL USP Clarithromycin Related Compound A RS and 125 g/mL clarithromycin from System suitability stock solution and Standard stock solution diluted with Mobile phase
100
USP 32
Mode: LC Detector: UV 210 nm Column: 4.6-mm 15-cm; 5-m packing L1 Temperature: 50 Flow rate: 1 mL/min Injection size: 5 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Determine the concentration, in mg/mL, of clarithromycin in the Sample solution at each time point: Result = (rU/rS) CS rU = peak response of the Sample solution rS = peak response of the Standard solution CS = concentration of the Standard solution (mg/mL) Calculate the amount, as a percentage, of clarithromycin dissolved with volume correction: Result = [{Cn [VM VU(n 1)]} + [ Ci VU]] 100/L = concentration of clarithromycin in the Sample solution at each time point (mg/mL) VM = volume of Medium, 900 mL VU = volume of the Sample withdrawn at each time point (mL) n = number of time points L = tablet label claim (mg) [NOTEThe summation of the amount of clarithromycin removed at previous sampling time points is applicable only where n is greater than 1.] Tolerances: The percentages of the labeled amounts of C38H69NO13 dissolved at the times specified conform to Acceptance Table 2.
Time (h) 2 12 24 Amount Dissolved (%) NMT 20 4570 NLT 80
Analysis Samples: Five Standard solutions and Sample solution Perform a linear regression analysis to generate a standard curve using the peak area of each Standard solution versus its concentration. Determine the amount in percentage of C38H69NO13 dissolved at each specified time interval, using the peak area of each Sample solution and the linear regression statistics for the Standard solutions. Tolerances: The percentages of the labeled amounts of C38H69NO13 dissolved at the times specified conform to the following Acceptance table.
Amount Dissolved, Individual Limits (%) NMT 65 5585 NLT 75 NLT 85 NMT 75 4595 NLT 65 NLT 75 NMT 2 tablets release more than 75%, and no individual tablet releases more than 85%. NMT 2 tablets are outside the range of 45%95%, and no individual tablet is outside the range of 35%105%. NMT 2 tablets release less than 65%, and no individual tablet releases less than 55%. NMT 2 tablets release less than 75%, and no individual tablet releases less than 65%. Amount Dissolved, Average Limits (%) NMT 65 5585 NLT 75 NLT 85 NMT 65
Level L1
Time (min) 30 45 60 120
L2
30 45 60 120
L3
30
Cn
45
5585
60
NLT 75
120
NLT 85
Test 2 [NOTEIf the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2.] Medium: 0.05 M phosphate buffer, pH 6.8 containing 0.5% of sodium lauryl sulfate; 900 mL, degassed by sonication and vacuum Apparatus 1: 100 rpm Times: 2, 12, and 24 h Determine the percentages of the labeled amount of C38H69NO13 dissolved using the following method. Solution A: 0.067 M Phosphate buffer, pH 2.5 (prepared by dissolving 9.2 g of monobasic sodium phosphate monohydrate in about 800 mL of water; adjust with phosphoric acid to a pH of 2.5; and dilute with water to 1000 mL) Mobile phase: Methanol and Solution A (13:7) Standard solution: 0.56 mg/mL of USP Clarithromycin RS in a solution of methanol and Medium (1:9) [NOTEInitially, sonicate to dissolve with methanol.] Sample solution: Centrifuge the solution under test at 2500 rpm for 10 min. Chromatographic system (See Chromatography 621, System Suitability.)
Test 3 [NOTEIf the product complies with this test, the labeling indicates that it meets USP Dissolution Test 3.] Medium: Acetate buffer, pH 4.75 (prepared by dissolving 3.59 g of sodium acetate trihydrate and 11.0 mL of 2 N acetic acid in 1000 mL of water; adjust with 2 N acetic acid to a pH of 4.75); 1000 mL Apparatus 1: 10 mesh; 50 rpm Times: 1, 2, 4, 8, and 12 h Determine the percentages of the labeled amount of C38H69NO13 dissolved by using the following method. Solution B: Dissolve 9.12 g of monobasic potassium phosphate in 1000 mL of water Mobile phase: Methanol and Solution B (13:7) Adjust with phosphoric acid to a pH of 4.0. Standard stock solution: 625 g/mL of USP Clarithromycin RS in methanol, taking into account the stated potency, in g/mg, of USP Clarithromycin RS Shake and sonicate, if necessary, to ensure dissolution.
USP 32
Standard solution: 125 g/mL of clarithromycin from Standard stock solution diluted with Mobile phase System suitability stock solution: 625 g/mL of USP Clarithromycin Related Compound A RS in methanol System suitability solution: 125 g/mL USP Clarithromycin Related Compound A and 125 g/mL of clarithromycin from System suitability stock solution and Standard stock solution diluted with Mobile phase Sample solution: Withdraw 10 mL of the solution under test. Transfer 3 mL to a 25-mL volumetric flask and dilute with Mobile phase to volume. Filter through a 0.45-m filter and replace 10 mL of Medium in each vessel. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 15-cm; 5-m packing L1 Column temperature: 50 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clarithromycin and clarithromycin related compound A are about 0.75 and 1.0, respectively, in System suitability solution.] Suitability requirements Resolution: NLT 2.0 between clarithromycin and clarithromycin related compound A, System suitability solution Column efficiency: NLT 750 theoretical plates from the clarithromycin peak, Standard solution Tailing factor: NLT 0.9 and NMT 2, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Determine the concentration, in mg/mL, of C38H69NO13 in the Sample solution at a time point: Result = (rU/rS) CS = peak response of the Sample solution rU = peak response of the Standard solution rS = concentration of the Standard solution (mg/mL) CS Calculate the amount, as a percentage, of clarithromycin dissolved with volume correction at time points n greater than 2: Result = [{Cn VM} + [ Ci VU]] 100/L = concentration of clarithromycin in the Sample solution at each time point (mg/mL) VM = volume of Medium, 1000 mL VU = volume of sample withdrawn at each time point (mL); n is the time point (at 2 h, n = 2), summation of the concentration of the Sample solution from the first to the (n 1)th time point (only applicable for n greater than 2) L = tablet label claim (mg) [NOTESummation of the concentration of the Sample solution from the first to the (n 1)th time point (only applicable for n equal to or greater than 2).] Tolerances: The percentages of the labeled amount of C38H69NO13 dissolved at the times specified conform to Acceptance Table 2. Cn
Time (h) 1 2 4 8 12

Amount Dissolved (%) NMT 15 1030 3555 NLT 80 NLT 90
Test 4 [NOTEIf the product complies with this test, the labeling indicates that it meets USP Dissolution Test 4.] Medium: Phosphate buffer, pH 6.0 (prepared by dissolving 6.8 mg/mL of potassium dihydrogen phosphate and 0.18 mg/mL of sodium hydroxide in water; adjust with dilute sodium hydroxide or phosphoric acid to a pH of 6.0 0.1); 900 mL Apparatus 2: 50 rpm Times: 2, 4, 8, and 12 h Determine the percentages of the labeled amount of C38H69NO13 dissolved by using the following method. Solution B: 6.8 mg/mL of potassium dihydrogen phosphate in water Adjust with dilute sodium hydroxide or phosphoric acid to a pH of 4.5 0.1. Mobile phase: Methanol and Solution B (16:9) Standard solution: Transfer 20 mg of USP Clarithromycin RS to a 50-mL volumetric flask. Dissolve in 30 mL of Medium, and sonicate until dissolved, about 10 min. Add 2 mL of methanol, and dilute with Medium to volume. Sample solution: Use the solution under test passed through a 0.45-m suitable filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 203 nm Column: 4.0-mm 12.5-cm; 5-m packing L7 Column temperature: 30 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Determine the concentration, in mg/mL of clarithromycin in the Sample solution at each time point: Result = (rU/rS) CS = peak response of the Sample solution rU = peak response of the Standard solution rS = concentration of the Standard solution (mg/mL) CS Calculate the amount, as a percentage, of clarithromycin dissolved at each time point: Result = Cn [VM (n 1) VS] + (C1 + C2 + . + Cn1) VS 100/L Cn VM VS = concentration in the Sample solution at each time point (mg/mL) = volume of Medium, 900 mL = volume of the sample taken at each time point (mL)
102
USP 32
System suitability solution: 0.5 mg/ml of amoxicillin dissolved in Standard solution Sample solution: 0.25 mg/mL of Clavulanate Lithium in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4-mm 30-cm; 310 m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clavulanic acid and amoxicillin are about 0.5 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.5 between the amoxicillin and clavulanic acid peaks in the System suitability solution Column efficiency: NLT 550 theoretical plates from the analyte peak in the Standard solution Tailing factor: NMT 1.5 for the analyte peak in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H9NO5 in each mg of the Clavulanate Potassium taken: Result = (rU/rS) (CS/CU) P F 100 peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (mg/mL) concentration of Clavulanate Postassium in the Sample solution (mg/mL) P = designated potency of USP Clavulanate Lithium, in g/mg of clavulanic acid F = unit conversion factor, 0.001mg/g Acceptance criteria: 75.5%92.0% IMPURITIES Organic Impurities PROCEDURE 1 Solution A: 0.05 M monobasic sodium phosphate, adjust with phosphoric acid to a pH of 4.0 0.1 Solution B: Methanol and Solution A (1:1) Mobile phase: Use variable mixtures of Solution A and Solution B as directed under Program Table
Program Table Time (min) 0 4 15 18 24 Solution A (%) 100 100 50 50 100 Solution B (%) 0 0 50 50 0
L = tablet label claim (mg) Tolerances: The percentages of the labeled amount of C38H69NO13 dissolved at the times specified conform to Acceptance Table 2.
Time (h) 2 4 8 12 Amount Dissolved (%) NMT 25 2040 4575 NLT 80
SPECIFIC TESTS LOSS ON DRYING 731: Dry a portion of powdered Tablets in vacuum at a pressure not exceeding 5 mm of mercury at 110 for 3 h: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, protected from light. Store at 25, excursions permitted between 15 and 30. LABELING: When more than one Dissolution test is given, the labeling states the Dissolution test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Clarithromycin RS USP Clarithromycin Related Compound A RS
Clavulanate Potassium
(Comment on this Monograph)id=m18000=Clavulanate Potassium=Chl-Cy-Monos.pdf)
rU rS CS CU
= = = =
237.25 C8H8KNO5 4-Oxa-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 3-(2hydroxyethylidene)-7-oxo-, monopotassium salt, 2R(2,3Z,5)-; Potassium (Z)-(2R,5R)-3-(2-hydroxyethylidene)-7-oxo-4-oxa-1azabicyclo[3.2.0]heptane-2-carboxylate [61177-45-5]. DEFINITION Clavulanate Potassium contains the equivalent of NLT 75.5% and NMT 92.0% of clavulanic acid (C8H9NO5), calculated on the anhydrous basis. IDENTIFICATION A. The retention time of the major peak for clavulanic acid in the Sample solution corresponds to that in the Standard solution, as obtained in the Assay. B. IDENTIFICATION TESTSGENERAL, Potassium 191: Meets the requirements ASSAY PROCEDURE Solution A: 7.8 mg/mL of monobasic sodium phosphate in water, adjust with phosphoric acid or 10 N sodium hydroxide to a pH of 4.4 0.1 Mobile phase: Methanol and Solution A (1:19) Standard solution: 0.25 mg/mL of USP Clavulanate Lithium RS in water
Standard solution: 0.1 mg/mL of USP Clavulanate Lithium RS in Solution A Sample solution: 10 mg/mL of Clavulanate Potassium in Solution A System suitability solution: 0.1 mg/mL each of USP Clavulanate Lithium RS and amoxicillin in Solution A Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 230 nm Column: 4.6-mm 10-cm; 5 m packing L1 Temperature: 40 Flow rate: 1 mL/min [NOTEThe system is equilibrated for 15 min with 100% Solution A] Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for clavulanic acid and amoxicillin are about 1.0 and 2.5, respectively.] Suitability requirements Resolution: NLT 13 between the clavulanic acid peak and the amoxicillin peak in the System suitability solution Column efficiency: NLT 2000 theoretical plates from the clavulanic acid peak in the System suitability solution Tailing factor: NMT 2.0 for the clavulanic acid peak in the System suitability solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage, in terms of clavulanate potassium equivalent, of each impurity in the Clavulanate Potassium taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response of an individual impurity peak from the Sample solution = clavulanic acid peak response from the Standard rS solution = concentration of Standard solution (mg/mL) CS = nominal concentration of Clavulanate Potassium CU from the Sample solution (mg/mL) Mr1 = molecular weight of clavulanate potassium, 237.3 Mr2 = molecular weight of clavulanate lithium, 205.1 Acceptance criteria Total impurities: NMT 2.0% PROCEDURE 2: LIMIT OF CLAVAM-2-CARBOXYLATE POTASSIUM Mobile phase: 0.1 M monobasic sodium phosphate, adjust with phosphoric acid to a pH of 4.0 0.1 Standard solution: 5 g/mL of USP Clavam-2-Carboxylate Potassium RS in water Sample solution: 10 mg/mL of Clavulanate Potassium in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4-mm 30-cm; 310 m packing L1 Flow rate: 0.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for clavam-2carboxylic acid and clavulanic acid are about 0.7 and 1.0, respectively.] Suitability requirements Column efficiency: NLT 4000 theoretical plates in the Standard solution Tailing factor: NMT 1.5 in the Standard solution Relative standard deviation: NMT 5.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of clavam-2-carboxylate potassium in the specimen taken: rU Result = (rU/rS) (CS/CU) 100 rU rS CS CU = = = =
Official Monographs / Clavulanate 103

peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (g/mL) concentration of Clavulanate Potassium in the Sample solution (g/mL) Acceptance criteria: NMT 0.01% PROCEDURE 3: LIMIT OF ALIPHATIC AMINES Internal standard solution: 50 L of 3-methyl-2pentanone in water to 100 mL Sample solution: Transfer 1.0 g of Clavulanate Potassium to a centrifuge tube, add 5.0 mL of Internal standard solution, 5.0 mL of 2 N sodium hydroxide, 5.0 mL of isopropyl alcohol, and 5 g of sodium chloride. Shake for 1 min, and centrifuge to separate the layers. Use the upper layer. Standard solution: Dissolve 80.0 mg of each of the following amines in 2 N hydrochloric acid: 1,1dimethylethylamine, diethylamine, tetramethylethylenediamine, 1,1,3,3-tetramethylbutylamine, and N,N-diisopropylethylenediamine. Dilute with 2 N hydrochloric acid to 200.0 mL. Transfer 5.0 mL of this solution to a centrifuge tube. Add 5.0 mL of the Internal standard solution, 10.0 mL of 2 N sodium hydroxide, 5.0 mL of isopropyl alcohol, and 5 g of sodium chloride. Shake for 1 min, and centrifuge to separate the layers. Use the upper layer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.53-mm 50-m capillary fused silica column that contains a 5-m film coating of stationary phase G41 Temperature Column temperature
Time (min) 0 7 10.8 125.8 Temperature () 35 35 150 150
Injector temperature: 200 Detector temperature: 250 Carrier gas: Helium Flow rate: 8 mL/min Split ratio: 1:10 Injection size: 1 L System suitability Sample: Standard solution [NOTEsee Table for relative retention times]
Name 1,1-Dimethylethylamine Diethylamine Isopropyl alcohol Tetramethylethylenediamine 1,1,3,3-Tetramethylbutylamine N,N-Diisopropylethylenediamine Bis(2-methylamino)ethyl ether Relative Retention Time 0.55 0.76 1.0 1.07 1.13 1.33 1.57
104
Clavulanate / Official Monographs

RU
USP 32
= peak response ratio of 2-ethylhexanoic acid to the 3-cyclohexylpropionic acid peak from the Sample solution = peak response ratio of 2-ethylhexanoic acid to RS the 3-cyclohexylpropionic acid peak from the Standard solution = concentration of 2-ethylhexanoic acid in the CS Standard solution (g/mL) CU = nominal concentration of Clavulanate Potassium in the Sample solution (g/mL) Acceptance criteria: NMT 0.8% SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: Where the label states that Clavulanate Potassium is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.03 USP Endotoxin Unit per mg. STERILITY TESTS 71: Where the label states that Clavulanate Potassium is sterile, it meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. PH 791: 5.58.0, in 10 mg/mL solution WATER DETERMINATION, Method I 921: NMT 1.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Clavam-2-Carboxylate Potassium RS USP Clavulanate Lithium RS USP Endotoxin RS
Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the Clavulanate Potassium taken: Result = (rU/rS) 100 = peak response for an individual impurity from the Sample solution = peak response for the relevant analyte from the rS Standard solution Calculate the percentage of any individual impurity for which no relevant reference compound is provided in the Standard solution by the same formula, except for rS use the peak response corresponding to the 1,1dimethylethylamine peak. Acceptance criteria Total of all aliphatic amines: NMT 0.2% PROCEDURE 4: LIMIT OF 2-ETHYLHEXANOIC ACID Internal standard solution: 1 mg/mL of 3cyclohexylpropionic acid in cyclohexane Standard solution: 1.5 mg/mL of 2-ethylhexanoic acid in Internal standard solution. Transfer 1.0 mL of this solution to a centrifuge tube, and add 4.0 mL of 4 N hydrochloric acid. Shake for 1 min, and allow the phases to separate, centrifuging if necessary. Withdraw the lower phase, and reserve the upper phase. To the lower phase add 1.0 mL of Internal standard solution, and shake for 1 min. Allow the phases to separate, centrifuging if necessary. Withdraw the upper phase, and combine with the reserved upper layer. Sample solution: Transfer 300 mg of Clavulanate Potassium to a centrifuge tube. Add 4.0 mL of 4 N hydrochloric acid, and shake with two successive 1.0 mL portions of the Internal standard solution. Allow the phases to separate, centrifuging if necessary. Use the combined upper phases. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.53-mm 25-m capillary fused silica column that contains a 1-m film coating of stationary phase G35 Temperature Column temperature rU
Time (min) 0 2 7.3 10.3 Temperature () 40 40 200 200
Clemastine Fumarate
(Comment on this Monograph)id=m18020=Clemastine Fumarate=Chl-Cy-Monos.pdf)
Injector temperature: 200 Detector temperature: 300 Carrier gas: Hydrogen Flow rate: 100 mL/min Injection size: 1 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between the 2-ethylhexanoic acid peak and the 3-cyclohexylpropionic acid peak in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of 2-ethylhexanoic acid in the Clavulanate Potassium taken: Result = (RU/RS) (CS/CU) 100
C21H26ClNO C4H4O4 459.96 Pyrrolidine, 2-[2-[1-(4-chlorophenyl)-1-phenylethoxy]ethyl]-1methyl-, [R-(R*,R*)]-, (E)-2-butenedioate (1:1); (+)-(2R)-2-[2-[[[(R)-p-Chloro--methyl--phenylbenzyl]oxy]ethyl]-1-methylpyrrolidine fumarate (1:1) [14976-57-9]. DEFINITION Clemastine Fumarate contains NLT 98.0% and NMT 102.0% of C21H26ClNO C4H4O4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 5 mg/mL of fumaric acid in dilute alcohol (8 in 10) Sample solution: 20 mg/mL of Clemastine Fumarate in dilute alcohol (8 in 10) with slight warming
USP 32
Spray reagent: 0.1 M potassium permanganate Application volume: 5 L Developing solvent system: Diisopropyl ether, formic acid, and water (14:5:1) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Dry the developed plate at 100 for 30 min, cool, and spray the plate with Spray reagent. Dry briefly with the aid of a current of warm air, and examine the chromatogram. Acceptance criteria: The principal spot of the Sample solution corresponds in RF value, color, and intensity to that of the Standard solution. ASSAY PROCEDURE Sample solution: 350 mg of Clemastine Fumarate in 60 mL of glacial acetic acid in a small conical flask Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 46.00 mg of C21H26ClNO C4H4O4. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities HEAVY METALS, METHOD II 231: NMT 20 ppm Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Diluent: Chloroform and methanol (1:1) Standard solution: 20 mg/mL of USP Clemastine Fumarate RS in Diluent Comparison solutions: Dilute portions of the Standard solution with the Diluent to prepare five Comparison solutions having known concentrations of 0.10, 0.08, 0.06, 0.04, and 0.02 mg/mL, respectively (0.5%, 0.4%, 0.3%, 0.2%, and 0.1% of the Standard solution, respectively). Sample solution: 20 mg/mL of Clemastine Fumarate in Diluent Solution A: 17 mg/mL of bismuth subnitrate in glacial acetic acid and water (1:4) Solution B: 0.4 g/mL of potassium iodide Spray reagent: Mix 5.0 mL of Solution A, 5.0 mL of Solution B, and 20 mL of glacial acetic acid in a 100mL volumetric flask, and dilute with water to volume. Application volume: 5 L Developing solvent system: Chloroform, methanol, and ammonium hydroxide (90:10:1) Analysis Samples: Standard solution, five Comparison solutions, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Locate the spots on the plate by spraying first with Spray reagent, then with 3% hydrogen peroxide: the principal spot of the Sample solution corresponds in RF value, color, and intensity to that of the Standard solution. Acceptance criteria Any individual impurity: NMT 0.5% Total impurities: NMT 1.0% SPECIFIC TESTS SPECIFIC ROTATION 781S: +15.0 to +18.0 at 20 Sample solution: 10 mg/mL, in methanol PH 791: 3.24.2, in a suspension (1 in 10) LOSS ON DRYING 731: Dry it at 105 to constant weight: it loses NMT 0.5% of its weight.
Official Monographs / Clemastine 105

CLARITY AND COLOR OF SOLUTION: Sample solution: 10 mg/mL of Clemastine Fumarate in methanol Comparison solution: Prepare a solution by mixing 2.5 mL of 0.02 mM in sodium chloride, 2.5 mL of water, 5.0 mL of 2.5 N nitric acid, and 1.0 mL of 0.1 N silver nitrate (2.5:2.5:5:1). [NOTEUse this solution within 5 min.] Color matching fluid: Matching Fluid C (see Color and Achromicity 631) and water (1:3) Analysis: Transfer 10 mL each of the Sample solution, the Comparison solution, and the Color matching fluid to separate test tubes having the same nominal diameter (about 12 mm). View the Sample solution and the Comparison solution horizontally against a dull black background: the Sample solution is clear or not more opalescent than the Comparison solution. View the Sample solution and the Color matching fluid horizontally against a dull white background: the Sample solution is colorless or not more intensely colored than the Color matching fluid. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at a temperature not exceeding 25. USP REFERENCE STANDARDS 11 USP Clemastine Fumarate RS
Clemastine Fumarate Tablets

(Comment on this Monograph)id=m18050=Clemastine Fumarate Tablets=Chl-Cy-Monos.pdf) DEFINITION Clemastine Fumarate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C21H26ClNO C4H4O4. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Diluent: Chloroform and methanol (1:1) Standard solution: 2.5 mg/mL of USP Clemastine Fumarate RS in Diluent Sample solution: Place a portion of powdered Tablets, equivalent to 2.5 mg of clemastine fumarate in a glassstoppered flask. Add 10 mL of Diluent, and shake for 20 min. Filter, wash the residue with two 5-mL portions of Diluent, and evaporate the combined filtrate and washings to dryness under vacuum. Dissolve the residue in 1 mL of Diluent. Solution A: 17 mg/mL of bismuth subnitrate in glacial acetic acid and water (1:4) Solution B: 0.4 g/mL of potassium iodide Spray reagent: Mix 5.0 mL of Solution A, 5.0 mL of Solution B, and 20 mL of glacial acetic acid in a 100-mL volumetric flask, and dilute with water to volume. Application volume: 5 L Developing solvent system: Chloroform, methanol, and ammonium hydroxide (90:10:1) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram until the solvent front has moved three-fourths of the length of the plate, mark the solvent front, and dry the plate at room temperature with the aid of a current of air. Locate the spots on the plate by spraying first with Spray reagent, then with 3% hydrogen peroxide.
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at the wavelength of maximum absorbance at about 420 nm of the chloroform solutions of the Sample solution and the Standard solution, using the chloroform solution of the blank to set the instrument. Tolerances: NLT 75% (Q) of the labeled amount of C21H26ClNO C4H4O4 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Solution A: 0.1 mg/mL of bromocresol purple in 0.33 N acetic acid Solution B: Methanol and 0.33 N acetic acid (1:9) Standard solution: 0.27 mg/mL of USP Clemastine Fumarate RS in Solution B. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with Solution B to volume. Sample solution: Mix 1 finely powdered Tablet with a volume of Solution B, sufficient to obtain 27 g/mL of clemastine fumarate solution. Shake for 30 min, filter, and discard the first few mL of the filtrate. Analysis: Transfer 15.0 mL each of the Standard solution, Sample solution, and Solution B to provide the blank to individual 125-mL separators. Add 25 mL of Solution A and 50.0 mL of chloroform to each, and shake by mechanical means for 15 min. Allow the layers to separate, and filter the chloroform layers. Concomitantly determine the absorbances of the filtered solutions of the Sample solution and the Standard solution at the wavelength of maximum absorbance at about 406 nm, using the blank to set the instrument. Calculate percentage of C21H26ClNO C4H4O4 in the Tablet taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Clemastine Fumarate RS in the Standard solution (g/mL) = nominal concentration of clemastine fumarate in the Sample solution (g/mL)
Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Solution A: 9.47 mg/mL of anhydrous dibasic sodium phosphate Solution B: 9.08 mg/mL of monobasic potassium phosphate Solution C: Solution A and Solution B (153:97) Solution D: Solution C and water (1:3) Mobile phase: Methanol and Solution D (83:17) Standard solution: 0.14 mg/mL of USP Clemastine Fumarate RS in methanol and water (1:1) Sample solution: Dilute a quantity equivalent to 14 mg of clemastine fumarate, from finely powdered Tablets (NLT 20), to a 200-mL conical flask. Pipet 100 mL of a mixture of methanol and water (1:1) into the flask, shake for 30 min, centrifuge, and filter the supernatant. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 4 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 1.5%, for 5 replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H26ClNO C4H4O4 in the portion of Tablets taken: Result= (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clemastine Fumarate RS in the Standard solution (mg/mL) = nominal concentration of clemastine fumarate in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Solution A: Dissolve 20.0 g of monohydrate citric acid in 1000 mL of water, add 22.0 mL of sodium hydroxide solution (3 in 10) and 8.8 mL of hydrochloric acid, and dilute with water to 2000 mL. Adjust, if necessary, with sodium hydroxide solution (1 in 2) to a pH of 4.0. Medium: Solution A; 500 mL Apparatus 2: 50 rpm Time: 30 min Sample solution: Centrifuge 60 mL of the solution under test for 20 min at 4000 rpm. Standard solution: Comparable concentration of USP Clemastine Fumarate RS in Dissolution Medium Analysis: Transfer 50.0 mL each of Sample solution, Standard solution, and Medium to three separate 125-mL separators, and treat each of the solutions as follows. Add 10 mL of methyl orange solution (2 in 10,000), mix, add 20.0 mL of chloroform, shake simultaneously by mechanical means for 10 min, remove the chloroform layer, and centrifuge the chloroform layer for 10 min at 4000 rpm. Determine the amount of C21H26ClNO C4H4O4 dissolved from absorbances
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clemastine Fumarate RS
Clidinium Bromide
(Comment on this Monograph)id=m18100=Clidinium Bromide=Chl-Cy-Monos.pdf)
C22H26BrNO3 432.35 1-Azoniabicyclo[2.2.2]octane, 3-[(hydroxydiphenylacetyl)oxy]-1methyl-, bromide, ()-; ()-3-Hydroxy-1-methylquinuclidinium bromide benzilate [3485-62-9]. DEFINITION Clidinium Bromide contains NLT 99.0% and NMT 100.5% of C22H26BrNO3, calculated on the dried basis.
USP 32
IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample: 250 mg Analysis: Dissolve the Sample in 5 mL of water in a test tube, cool in an ice bath, add 5 mL of trinitrophenol TS, and scratch the inner surface of the tube with a glass rod to induce crystallization. Collect the precipitate on a filter, wash with cold water, and dry at 105 for 1 h. Test the picrate so obtained by the method for Class I (see Melting Range or Temperature 741). [CAUTIONPicrates may explode.] Acceptance criteria: The picrate melts between 184 and 189. C. PROCEDURE Sample solution: 50 mg/mL Analysis: To the Sample solution, add a few drops of 2 N nitric acid and 1 mL of silver nitrate TS. Acceptance criteria: A yellowish white precipitate is formed. ASSAY PROCEDURE Sample: 1.2 g Analysis: Dissolve the Sample in 80 mL of glacial acetic acid, warming if necessary to effect solution. Cool, and add 15 mL of mercuric acetate TS. Titrate with 0.1 N perchloric acid in dioxane VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 43.24 mg of C22H26BrNO3. Acceptance criteria: 99.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS 231 Sample solution: 40 mg/mL Acceptance criteria: NMT 20 ppm Organic Impurities PROCEDURE Standard solution: 100 mg/mL of USP Clidinium Bromide RS in 0.1 N methanolic hydrochloric acid Sample solution: 100 mg/mL of Clidinium Bromide in 0.1 N methanolic hydrochloric acid Reference solution 1: Dissolve 3.0 mg of USP 3Quinuclidinyl Benzilate RS in 100 mL of 0.1 N methanolic hydrochloric acid. Reference solution 2: Dissolve 100 mg of USP Clidinium Bromide RS in 1.0 mL of 0.1 N methanolic hydrochloric acid, and add 20 L of a solution of 25.0 mg of USP Clidinium Bromide Related Compound A RS in 1.0 mL of 0.1 N methanolic hydrochloric acid. Spray reagent: Dissolve 850 mg of bismuth subnitrate in a mixture of 10 mL of glacial acetic acid and 40 mL of water. In a separate container, dissolve 20 g of potassium iodide in 50 mL of water. Mix the two solutions, and dilute with dilute sulfuric acid (1 in 10) to 500 mL. Add 7.5 g 2.5 g of iodine, and mix until the solution is complete. Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Developing solvent system: Acetone, methanol, hydrochloric acid, and water (70:20:5:5) Chromatographic plates: (See Chromatography 621, Thin-Layer Chromatography.) Predevelop suitable thin-layer chromatographic plates by placing in a chromatographic chamber saturated with Developing solvent system, and allow the Developing solvent system to move about 15 cm. Remove the plates from the chamber, dry at 105 for 15 min, and cool.
Official Monographs / Clindamycin 107

Analysis 1 (3-quinuclidinyl benzilate): Proceed as directed under Chromatography 621, Thin-Layer Chromatography, applying the Standard solution, Reference solution 1, and the Sample solution. Place the plate in an unsaturated chromatographic chamber containing freshly prepared Developing solvent system, and allow the solvent front to move 10 cm. Remove the plate, dry at 105 for 10 min, cool, and spray with potassium iodoplatinate TS. Acceptance criteria 1: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution, and the Sample solution shows no spot at an RF value (about 0.8) corresponding to that of 3-quinuclidinyl benzilate. Analysis 2 (limit of clidinium bromide related compound A): Proceed as directed under Chromatography 621, Thin-Layer Chromatography, using Reference solution 2 and the Sample solution. Place the plate in an unsaturated chromatographic chamber containing freshly prepared Developing solvent system, and allow the solvent front to move 15 cm. Remove the plate, dry at 105 for 10 min, cool, and spray with Spray reagent. Acceptance criteria 2: Any spot in the chromatogram of the Sample solution at an RF value of about 0.4 is not greater in size or intensity than the minor spot of Reference solution 2: NMT 0.5% of clidinium bromide related compound A. SPECIFIC TESTS LOSS ON DRYING 731 Analysis: Dry a sample at 105 for 3 h. Acceptance criteria: The sample loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clidinium Bromide RS USP Clidinium Bromide Related Compound A RS USP 3-Quinuclidinyl Benzilate RS
Clindamycin Hydrochloride
(Comment on this Monograph)id=m18160=Clindamycin Hydrochloride=Chl-Cy-Monos.pdf)
461.45 C18H33ClN2O5S HCl L-threo--D-galacto-Octopyranoside, methyl 7-chloro-6,7,8trideoxy-6-[[(1-methyl-4-propyl-2-pyrrolidinyl)carbonyl]amino]-1-thio-, (2S-trans)-, monohydrochloride; Methyl 7-chloro-6,7,8-trideoxy-6-(1-methyl-trans-4-propyl-L-2pyrrolidinecarboxamido)-1-thio-L-threo--D-galactooctopyranoside monohydrochloride [21462-39-5]. Monohydrate [58207-19-5]. 479.47 DEFINITION Clindamycin Hydrochloride is the hydrated hydrochloride salt of clindamycin, a substance produced by the chlorination of lincomycin. It has a potency equivalent to NLT 800 g of C18H33ClN2O5S/mg.
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Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; 5 m packing L1 Flow rate: 1.0 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements [NOTEThe relative retention times are 0.4 for lincomycin, 0.65 for clindamycin B, 0.8 for 7epiclindamycin, and 1.0 for clindamycin.] Analysis Samples: Standard solution and Sample solution Record the chromatograms for a period of time that is six times the retention time of clindamycin. Calculate the percentage of lincomycin in the Clindamycin Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Lincomycin Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of lincomycin CU hydrochloride in the Sample solution (mg/mL) Calculate the percentage of all other related compounds in the Clindamycin Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = response of an individual related compound, other than lincomycin, in the Sample solution = clindamycin peak response in the Standard rS solution = concentration of USP Clindamycin CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria Individual impurities: See the Impurity Table. Total of all related compounds, including lincomycin: NMT 6.0% rU
Impurity Table Impurity Name 7-Epiclindamycin Clindamycin B Any other individual related compound Acceptance Criteria, NMT (%) 4.0 2.0 1.0
IDENTIFICATION INFRARED ABSORPTION 197M:
ASSAY PROCEDURE Solution A: 6.8 mg/mL of monobasic potassium phosphate in water, and adjust with 8 N potassium hydroxide to a pH of 7.5. Mobile phase: Acetonitrile and Solution A (9:11) [NOTEIncreasing the proportion of acetonitrile in the Mobile phase decreases the retention time, and decreasing it increases the resolution between 7-epiclindamycin and clindamycin.] Standard solution: 1 mg/mL of USP Clindamycin Hydrochloride RS in Mobile phase Sample solution: 1 mg/mL of Clindamycin Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; 5m packing L1 Flow rate: 1.0 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.4 between the clindamycin B peak and the 7-epiclindamycin peak, and NLT 3.0 between the 7epiclindamycin peak and the clindamycin peak Column efficiency: NLT 4000 theoretical plates from the clindamycin peak Tailing factor: NMT 1.2 for the clindamycin peak Relative standard deviation: NMT 1.0% for the clindamycin peak Analysis Samples: Standard solution and Sample solution Record the chromatograms for a period of time that is twice the retention time of the clindamycin peak. Calculate the potency of C18H33ClN2O5S in g/mg, in the portion of Clindamycin Hydrochloride taken: Result = (rU/rS) (CS/CU) P = peak area response from the Sample solution = peak area response from the Standard solution = concentration of USP Clindamycin Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of clindamycin hydrochloride in the Sample solution (mg/mL) P = potency of clindamycin (g/mg of USP Clindamycin Hydrochloride RS) Acceptance criteria: NLT 800 g/mg IMPURITIES Organic Impurities PROCEDURE Solution A and Mobile phase: Prepare as directed in the Assay. Standard stock solution: 0.5 mg/mL of USP Lincomycin Hydrochloride RS and 1 mg/mL of USP Clindamycin Hydrochloride RS in Mobile phase Standard solution: Standard stock solution and Mobile phase (1:9) Sample solution: 5 mg/mL of Clindamycin Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) rU rS CS
rU rS CS
SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 3.05.5, in a 100 mg/mL solution WATER DETERMINATION, Method I 921: 3.0%6.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clindamycin Hydrochloride RS USP Lincomycin Hydrochloride RS
USP 32

Apparatus 1: 100 rpm Time: 30 min Solution A: Dissolve 16 g of dl-10-camphorsulfonic acid, 8 g of ammonium acetate, and 8 mL of glacial acetic acid in 1600 mL of water. Mobile phase: Methanol and Solution A (3:2). Adjust with hydrochloric acid or 5 N sodium hydroxide to a pH of 6.0 0.05. Standard solution: Prepare a solution of USP Clindamycin Hydrochloride RS in Medium having a known concentration similar to that expected in the Sample solution. Sample solution: Use a filtered portion of the Sample solution, diluted with Medium if necessary. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index detector Column: 3 m, packing L1 Flow rate: 2.0 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the amount of C18H33ClN2O5S dissolved. Acceptance criteria: NLT 80% (Q) of the labeled amount of C18H33ClN2O5S is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 7.0%
Clindamycin Hydrochloride Capsules

(Comment on this Monograph)id=m18170=Clindamycin Hydrochloride Capsules=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Hydrochloride Capsules contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of clindamycin (C18H33ClN2O5S). IDENTIFICATION The retention time of the major peak of the Sample solution obtained as directed in the Assay corresponds to that of the Standard solution, relative to the internal standard. ASSAY PROCEDURE Solution A: Add 2 g of dl-10-camphorsulfonic acid, 1 g of ammonium acetate, and 1 mL of glacial acetic acid to 200 mL of water. Mobile phase: Methanol and Solution A (3:2) Adjust, if necessary, with hydrochloric acid or a sodium hydroxide solution (1 in 2) to a pH of 6.0 0.1. Internal standard solution: Phenylethyl alcohol and Mobile phase (0.5:99.5) Standard solution: 18 mg/mL of USP Clindamycin Hydrochloride RS in Internal standard solution Sample solution: Remove as completely as possible the contents of NLT 20 Capsules, weigh, and mix. Transfer equivalent to 75 mg of clindamycin from powder to a suitable container. Add 5.0 mL of Internal standard solution, and shake for 30 min. Centrifuge or filter, if necessary, to obtain a clear solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index detector Column: 4-mm 30-cm stainless steel column; packing L1 Flow rate: 1.0 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for the internal standard and clindamycin are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 5.0 between the analyte and internal standard Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H33ClN2O5S in the portion of Capsules taken: Result = (rU/rS) (CS/CU) P 100 = peak response ratio of clindamycin to the internal standard from the Sample solution = peak response ratio of clindamycin to the rS internal standard from the Standard solution CS = concentration of USP Clindamycin Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of clindamycin in the CU Sample solution (mg/mL) P = potency of clindamycin (g/mg of USP Clindamycin Hydrochloride) Acceptance criteria: 90.0%120.0% rU PERFORMANCE TESTS DISSOLUTION 711 Medium: pH 6.8 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions); 900 mL
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clindamycin Hydrochloride RS
Clindamycin Hydrochloride Oral Solution

(Comment on this Monograph)id=m18180=Clindamycin Hydrochloride Oral Solution=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Hydrochloride Oral Solution contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of clindamycin (C18H33ClN2O5S). IDENTIFICATION The retention time of the clindamycin peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 6.8 mg/mL of monobasic potassium phosphate in water, and adjust with 8 N potassium hydroxide to a pH of 7.5 Mobile phase: Acetonitrile and Solution A (9:11) [NOTEIncreasing the proportion of acetonitrile in the Mobile phase decreases the retention time, and decreasing it increases the resolution between 7-epiclindamycin and clindamycin.] Standard solution: 1 mg/mL of USP Clindamycin Hydrochloride RS in Mobile phase
110

IDENTIFICATION INFRARED ABSORPTION 197M:
USP 32
Sample solution: Equivalent to 0.85 mg/mL of clindamycin from Oral Solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.4 between the clindamycin B and 7epiclindamycin peaks, and NLT 3.0 between the 7epiclindamycin and clindamycin peaks Column efficiency: NLT 4000 theoretical plates from the clindamycin peak Tailing factor: NMT 1.2 for the clindamycin peak Relative standard deviation: NMT 1.0% for the clindamycin peak Analysis Samples: Standard solution and Sample solution Record the chromatograms for a period of time that is twice the retention time of the clindamycin peak. Calculate the percentage of C18H33ClN2O5S in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) P 100 = peak area response from the Sample solution = peak area response from the Standard solution = concentration of USP Clindamycin Hydrochloride in the Standard solution (mg/mL) = nominal concentration of clindamycin CU hydrochloride in the Sample solution (mg/mL) P = potency of clindamycin (g/mg of USP Clindamycin Hydrochloride) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For solution packaged in single-unit containers: meets the requirements SPECIFIC TESTS PH 791: 2.56.0 DELIVERABLE VOLUME 698: rU rS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label Oral Solution to indicate that it is intended for veterinary use only. USP REFERENCE STANDARDS 11 USP Clindamycin Hydrochloride RS
ASSAY PROCEDURE Internal standard solution: 5 mg/mL of cholesteryl benzoate in chloroform Standard solution: Transfer 150 mg of USP Clindamycin Palmitate Hydrochloride RS to a glass-stoppered, 15-mL conical centrifuge tube. Add 5 mL of water, 5.0 mL of Internal standard solution, and 1 mL of sodium carbonate solution (3 in 10). Insert the stopper, shake vigorously for NLT 10 min, and centrifuge. Remove the upper aqueous layer, and transfer 1.0 mL of the lower chloroform layer to a 15-mL centrifuge tube. Add 1.0 mL of pyridine and 1.0 mL of acetic anhydride. Agitate the tube to ensure complete mixing, cover the top of the centrifuge tube with a plastic cap through which a small hole has been punched, heat at 100 for 2.5 h, and allow to cool. Mix, and centrifuge, if necessary. Use the clear solution. Sample solution: Transfer 150 mg of Clindamycin Palmitate Hydrochloride to a glass-stoppered, 15-mL conical centrifuge tube. Add 5 mL of water, 5.0 mL of Internal standard solution, and 1 mL of sodium carbonate solution (3 in 10). Insert the stopper, shake vigorously for NLT 10 min, and centrifuge. Remove the upper aqueous layer, and transfer 1.0 mL of the lower chloroform layer to a 15-mL centrifuge tube. Add 1.0 mL of pyridine and 1.0 mL of acetic anhydride. Agitate the tube to ensure complete mixing, cover the top of the centrifuge tube with a plastic cap through which a small hole has been punched, heat at 100 for 2.5 h, and allow to cool. Mix, and centrifuge, if necessary. Use the clear solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.6-m 3-mm glass column packed with 1% phase G36 on support S1AB Column temperature: 290 Detector temperature: 320 Carrier gas: Dry helium Flow rate: 60 mL/min Injection size: 1.0 L Analysis Samples: Standard solution and Sample solution In a suitable chromatogram, the resolution of the peaks is complete. The elution order is: cholesteryl benzoate, clindamycin palmitate. Calculate the potency, in g of C18H33ClN2O5S/mg, in the Clindamycin Palmitate Hydrochloride taken: Result = (RU/RS) (CS/CU) P RU RS CS CU P = peak response ratio of clindamycin palmitate to cholesteryl benzoate from the Sample solution = peak response ratio of clindamycin palmitate to cholesteryl benzoate from the Standard solution = concentration of USP Clindamycin Palmitate Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of Clindamycin Palmitate Hydrochloride in the Sample solution (mg/mL) = potency of the USP Clindamycin Palmitate Hydrochloride RS, in g of clindamycin/mg
Clindamycin Palmitate Hydrochloride

(Comment on this Monograph)id=m18220=Clindamycin Palmitate Hydrochloride=Chl-Cy-Monos.pdf) C34H63ClN2O6S HCl 699.85 L-threo-- D-galacto-Octopyranoside, methyl 7-chloro-6,7,8trideoxy-6-[[(1-methyl-4-propyl-2-pyrrolidinyl)carbonyl]amino]-1-thio-2-hexadecanoate, monohydrochloride, (2S-trans)-; Methyl 7-chloro-6,7,8-trideoxy-6-(1-methyl-trans-4-propyl-L-2pyrrolidinecarboxamido)-1-thio-L-threo-- D-galactooctopyranoside 2-palmitate monohydrochloride [25507-04-4]. DEFINITION Clindamycin Palmitate Hydrochloride has a potency equivalent to NLT 540 g of clindamycin (C18H33ClN2O5S)/mg.
USP 32
Acceptance criteria: NLT 540 g

Column temperature: 290 Detector temperature: 320 Carrier gas: Dry helium Flow rate: 60 mL/min Injection size: 1 L Analysis Samples: Standard solution and Sample solution In a suitable chromatogram, the resolution of the peaks is complete. The elution order is cholesteryl benzoate, clindamycin palmitate. Calculate the percentage of C18H33ClN2O5S in each mL of the solution constituted from Clindamycin Palmitate Hydrochloride for Oral Solution taken: Result = (RU/RS) (CS/CU) P 100
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
NMT 0.5%
SPECIFIC TESTS PH 791: 2.83.8, in a 10 mg/mL solution WATER DETERMINATION, Method I 921: NMT 3.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clindamycin Palmitate Hydrochloride RS
Clindamycin Palmitate Hydrochloride for Oral Solution

(Comment on this Monograph)id=m18250=Clindamycin Palmitate Hydrochloride for Oral Solution=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Palmitate Hydrochloride for Oral Solution is a dry mixture of Clindamycin Palmitate Hydrochloride and one or more suitable buffers, colors, diluents, flavors, and preservatives. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of clindamycin (C18H33ClN2O5S), the labeled amount being 15 mg/mL when constituted as directed in the labeling. ASSAY PROCEDURE Internal standard solution: 5 mg/mL of cholesteryl benzoate in chloroform Standard solution: Transfer 150 mg of USP Clindamycin Palmitate Hydrochloride RS to a glass-stoppered, 15-mL conical centrifuge tube. Add 5 mL of water, 5.0 mL of Internal standard solution, and 1 mL of sodium carbonate solution (3 in 10). Insert the stopper, shake vigorously for NLT 10 min, and centrifuge. Remove the upper aqueous layer, and transfer 1.0 mL of the lower chloroform layer to a 15-mL centrifuge tube. Add 1.0 mL of pyridine and 1.0 mL of acetic anhydride. Agitate the tube to ensure complete mixing, cover the top of the centrifuge tube with a plastic cap through which a small hole has been punched, heat at 100 for 2.5 h, and allow to cool. Mix and centrifuge, if necessary. Use the clear solution. Sample solution: Constitute the Clindamycin Palmitate Hydrochloride for Oral Solution as directed in the labeling, and transfer 5.0 mL of the constituted solution to a glassstoppered, 15-mL conical centrifuge tube. Add 5.0 mL of Internal standard solution and 1 mL of sodium carbonate solution (3 in 10). Insert the stopper, shake vigorously for NLT 10 min, and centrifuge. Remove the upper aqueous layer, and transfer 1.0 mL of the lower chloroform layer to a 15-mL centrifuge tube. Add 1.0 mL of pyridine and 1.0 mL of acetic anhydride. Agitate the tube to ensure complete mixing, cover the top of the centrifuge tube with a plastic cap through which a small hole has been punched, heat at 100 for 2.5 h, and allow to cool. Mix and centrifuge, if necessary. Use the clear solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.6-m 3-mm glass column packed with 1% phase G36 on support S1AB
= peak response ratio of clindamycin palmitate to the cholesteryl benzoate peak from the Sample solution = peak response ratio of clindamycin palmitate to RS the cholesteryl benzoate peak from the Standard solution = concentration of USP Clindamycin Palmitate CS Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of clindamycin palmitate CU hydrochloride in the Sample solution (mg/mL) P = potency of the USP Clindamycin Palmitate Hydrochloride RS (g of clindamycin/mg) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 (for solid packaged in single-unit containers): Meets the requirements SPECIFIC TESTS PH 791: 2.55.0, in the solution constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 3.0% DELIVERABLE VOLUME 698: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clindamycin Palmitate Hydrochloride RS
RU
Clindamycin Phosphate
(Comment on this Monograph)id=m18280=Clindamycin Phosphate=Chl-Cy-Monos.pdf) 504.97 C18H34ClN2O8PS L-threo--D-galacto-Octopyranoside, methyl 7-chloro-6,7,8trideoxy-6-[[(1-methyl-4-propyl-2-pyrrolidinyl)carbonyl]amino] -1-thio-, 2-(dihydrogen phosphate), (2S-trans)-; Methyl 7-chloro-6,7,8-trideoxy-6-(1-methyl-trans-4-propyl-L-2pyrrolidinecarboxamido)-1-thio-L-threo--D-galactooctopyranoside 2-(dihydrogen phosphate) [24729-96-2]. DEFINITION Clindamycin Phosphate has a potency equivalent to NLT 758 g of C18H33ClN2O5S/mg, calculated on the anhydrous basis.
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Acceptance criteria: NLT 758 g/mg
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IDENTIFICATION INFRARED ABSORPTION 197M Sample: Undried ASSAY PROCEDURE Solution A: Add 14 mL of phosphoric acid to 4000 mL of HPLC grade water. Add 10 mL of ammonium hydroxide, and adjust with ammonium hydroxide to a pH of 3.90 0.05. Solution B: Acetonitrile and methanol (9:1) Diluent: Solution B and Solution A (1:4) Solution C: Solution B and Solution A (2:23) Solution D: Solution B and Solution A (12:13) Mobile phase: Use variable mixtures of Solution C and Solution D as directed under Chromatographic system. Standard solution: Weigh 22 mg of USP Clindamycin Phosphate RS. Add 10.0 mL of Diluent, shake briefly, and sonicate for 5 min to dissolve. Allow to cool to ambient temperature. Sample solution: Weigh 22 mg of Clindamycin Phosphate. Add 10.0 mL of Diluent, shake briefly, and sonicate for 5 min to dissolve. Allow to cool to ambient temperature. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; packing L7 Temperature: 40 Flow rate: 1.2 mL/min
Program Table Time (min) 0 40 41 46 Solution A (%) 95.0 5.0 95.0 95.0 Solution B (%) 5.0 95.0 5.0 5.0
SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 3.54.5, in a 10 mg/mL solution WATER DETERMINATION, Method I 921: NMT 6.0% STERILITY TESTS 71: Meets the requirements when tested as directed under Test for Sterility of the Product to be Examined, Membrane Filtration, and using 6 g of specimen aseptically dissolved in 200 mL of Fluid A. BACTERIAL ENDOTOXINS 85: Where the label states that Clindamycin Phosphate must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements. It contains NMT 0.58 USP Endotoxin Unit/mg of clindamycin. OTHER REQUIREMENTS: Where the label states that Clilndamycin Phosphate is sterile, it meets the requirements. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Clindamycin Phosphate RS USP Endotoxin RS
Clindamycin Phosphate Vaginal Cream

(Comment on this Monograph)id=m18283=Clindamycin Phosphate Vaginal Cream=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Phosphate Vaginal Cream contains an amount of clindamycin phosphate equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of clindamycin (C18H33ClN2O5S). IDENTIFICATION The relative retention time of the major peak for clindamycin phosphate of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 13.6 mg/mL of monobasic potassium phosphate in water, adjust with phosphoric acid to a pH of 2.5 Mobile phase: Acetonitrile and Solution A (9:31) System suitability solution: 0.6 mg/mL each of USP Clindamycin Phosphate RS and USP Clindamycin Hydrochloride RS in the Mobile phase Standard solution: 0.25 mg/mL of USP Clindamycin Phosphate RS, in the Mobile phase Sample solution: Transfer an equivalent to 20 mg of clindamycin from Cream to a 125-mL conical flask, add 100.0 mL of Mobile phase and 10 glass beads (10 mm in diameter). Insert the stopper in the flask securely, and shake by mechanical means at 50 for 1 h. Cool in an ice bath for 20 min, and centrifuge. Filter a few mL of the cloudy lower layer through a filter of 2 m or finer porosity, and use the filtrate. Chromatographic system (See Chromatography 621, System Suitability.)
Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.0 between clindamycin phosphate and its nearest related compound, determined as follows: Between the peaks, draw a line perpendicular to the base line at the valley that separates the two peaks. The height of the valley from the base line is NMT 40% of the height of the related compound peak. Calculate the resolution, R, using peak widths at half height. Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity of C18H33ClN2O5S, in g, in the portion of Clindamycin Phosphate taken: Result = (rU/rS) (CS/CU) P rU rS CS CU P = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) = nominal concentration of clindamycin phosphate in the Sample solution (mg/mL) = potency of clindamycin in the USP Clindamycin Phosphate RS (g/mg)
USP 32
Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for clindamycin phosphate and clindamycin are about 1.0 and 1.5, respectively.] Suitability requirements Resolution: NLT 6.0 between the clindamycin phosphate peak and the clindamycin peak, System suitability solution Column efficiency: NLT 1700 theoretical plates, System suitability solution Tailing factor: NMT 1.3, System suitability solution Relative standard deviation: NMT 2.5%, Standard solution Analysis Samples: Standard solution and Sample solution. Calculate the percentage of C18H33ClN2O5S in the portion of Cream taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) = nominal concentration of clindamycin phosphate CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 3.06.0, determined on the undiluted Cream ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clindamycin Hydrochloride RS USP Clindamycin Phosphate RS rU rS CS

means for 30 min. Centrifuge a portion of the solution thus obtained, and if necessary, filter a portion of the supernatant. Use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clindamycin phosphate and clindamycin are 1.0 and 1.5, respectively.] Suitability requirements Resolution: NLT 6.0 between the clindamycin phosphate and the clindamycin peaks, System suitability solution Column efficiency: NLT 1700 theoretical plates, in the System suitability solution, when calculated: 5.545 (tr/Wh/2)2 Tailing factor: NMT 1.3, System suitability solution Relative standard deviation: NMT 2.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H33ClN2O5S in the portion of Gel taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) = nominal concentration of clindamycin phosphate CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 4.56.5 MINIMUM FILL 755:
Clindamycin Phosphate Gel

(Comment on this Monograph)id=m18285=Clindamycin Phosphate Gel=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Phosphate Gel contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of clindamycin (C18H33ClN2O5S). IDENTIFICATION The retention time of the clindamycin phosphate peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 13.6 mg/mL of monobasic potassium phosphate in water, adjust with phosphoric acid to a pH of 2.5 Mobile phase: Acetonitrile and Solution A (9:31) System suitability solution: 0.6 mg/mL each of USP Clindamycin Phosphate RS and USP Clindamycin Hydrochloride RS in Mobile phase Standard solution: 0.25 mg/mL of USP Clindamycin Phosphate RS, in Mobile phase Sample solution: Transfer an equivalent to 20 mg of clindamycin from Gel to a 100-mL volumetric flask, dilute with Mobile phase to volume, and shake by mechanical
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clindamycin Hydrochloride RS USP Clindamycin Phosphate RS
Clindamycin Injection
(Comment on this Monograph)id=m18289=Clindamycin Injection=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Injection contains an amount of Clindamycin Phosphate in Water for Injection equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of clindamycin (C18H33ClN2O5S). It may be frozen. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 13.6 mg/mL of monobasic potassium phosphate. Adjust with phosphoric acid to a pH of 2.5.
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Mobile phase: Acetonitrile and Solution A (9:31) [NOTEEnsure that the concentration of acetonitrile in the Mobile phase is NLT 22% and NMT 25% in order to retain the correct elution order.] System suitability solution: 25 g/mL of USP Benzyl Alcohol RS and 250 g/mL of USP Clindamycin Phosphate RS, in Mobile phase Standard solution: 0.24 mg/mL of USP Clindamycin Phosphate RS, in Mobile phase Sample stock solution: Transfer a volume of Injection, equivalent to 300 mg of clindamycin, to a 100-mL volumetric flask, dilute with Mobile phase to volume, and mix. Sample solution: Sample stock solution and Mobile phase (7:93), 0.21 mg/mL of clindamycin Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clindamycin phosphate and benzyl alcohol are 1.0 and 1.2, respectively.] Suitability requirements Resolution: NLT 2.0 between clindamycin phosphate and benzyl alcohol, System suitability solution Relative standard deviation: NMT 2.5, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H33ClN2O5S in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of clindamycin phosphate from the Sample solution = peak response of clindamycin phosphate from rS the Standard solution CS = concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) CU = nominal concentration of clindamycin phosphate in the Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% rU SPECIFIC TESTS PH 791: 5.57.0 BACTERIAL ENDOTOXINS TEST 85: Contains NMT 0.58 USP Endotoxin Unit/mg of clindamycin. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections. OTHER REQUIREMENTS: Meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, or in suitable plastic containers. LABELING: Meets the requirements under Injections 1, Labeling. Where it is maintained in the frozen state, the label states that it is to be thawed just prior to use, describes the conditions for proper storage of the resultant solution, and directs that the solution is not to be refrozen. USP REFERENCE STANDARDS 11 USP Benzyl Alcohol RS USP Clindamycin Phosphate RS USP Endotoxin RS
Clindamycin for Injection

(Comment on this Monograph)id=m18301=Clindamycin for Injection=Chl-Cy-Monos.pdf) DEFINITION Clindamycin for Injection contains an amount of Clindamycin Phosphate having a potency equivalent to NLT 758 g of clindamycin (C18H33ClN2O5S)/mg, calculated on the anhydrous basis. IDENTIFICATION INFRARED ABSORPTION 197M Sample specimen: Undried ASSAY PROCEDURE Solution A: Add 14 mL of phosphoric acid to 4000 mL of HPLC grade water. Add 10 mL of ammonium hydroxide, and adjust with ammonium hydroxide to a pH of 3.90 0.05. Solution B: Acetonitrile and methanol (9:1) Diluent: Solution B and Solution A (1:4) Solution C: Solution B and Solution A (2:23) Solution D: Solution B and Solution A (12:13) Mobile phase: Use variable mixtures of Solution C and Solution D as directed for Chromatographic system Standard solution: Weigh 22 mg of USP Clindamycin Phosphate RS. Add 10.0 mL of Diluent, shake briefly, and sonicate for 5 min to dissolve. Allow to cool to ambient temperature. Sample solution: Weigh 22 mg of Clindamycin Phosphate. Add 10.0 mL of Diluent, shake briefly, and sonicate for 5 min to dissolve. Allow to cool to ambient temperature. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; packing L7 Temperature: 40 Flow rate: 1.2 mL/min
Program Table Time (min) 0 40 41 46 Solution A (%) 95.0 5.0 95.0 95.0 Solution B (%) 5.0 95.0 5.0 5.0
Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.0 between clindamycin phosphate and its nearest related compound, determined as follows: Between the peaks, draw a line perpendicular to the base line at the valley that separates the two peaks. The height of the valley from the base line is NMT 40% of the height of the related compound peak. Calculate the resolution, R, using peak widths at half height. Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity of C18H33ClN2O5S, in g, in the portion of Injection taken: Result = (rU/rS) (CS/CU) P
USP 32
= peak area from the Sample solution = peak area from the Standard solution = concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) = nominal concentration of clindamycin phosphate CU in the Sample solution (mg/mL) P = potency of clindamycin in the USP Clindamycin Phosphate RS (g/mg) Acceptance criteria: NLT 758 g/mg SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: Contains NMT 0.58 USP Endotoxin Unit/mg of clindamycin. STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration, 6 g of specimen aseptically dissolved in 200 mL of Fluid A being used. PH 791: 3.54.5, in 10 mg/mL solution WATER DETERMINATION, Method I 921: NMT 6.0% CRYSTALLINITY 695: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Clindamycin Phosphate RS rU rS CS

Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clindamycin phosphate and 4-hydroxyacetophenone are about 1.0 and 1.2, respectively.] Suitability requirements Resolution: NLT 2.0 between clindamycin phosphate and 4-hydroxyacetophenone, System suitability solution Relative standard deviation: NMT 2.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H33ClN2O5S in each mL of the Topical Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) CU = nominal concentration of clindamycin phosphate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.07.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clindamycin Phosphate RS rU rS CS
Clindamycin Phosphate Topical Solution

(Comment on this Monograph)id=m18303=Clindamycin Phosphate Topical Solution=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Phosphate Topical Solution contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of clindamycin (C18H33ClN2O5S). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 13.6 mg/mL of monobasic potassium phosphate in water, adjust with phosphoric acid to a pH of 2.5 Mobile phase: Acetonitrile and Solution A (9:31) [NOTEEnsure that the concentration of acetonitrile in the Mobile phase is NLT 22% and NMT 25% in order to retain the correct elution order.] Standard solution: 0.24 mg/mL of USP Clindamycin Phosphate RS in Mobile phase System suitability stock solution: 4 mg/mL of 4hydroxyacetophenone in acetonitrile System suitability solution: Prepare 0.04 mg/mL of 4hydroxyacetophenone from this solution with Mobile phase. Mix the resulting solution and Standard solution (1:3). Sample solution: Equivalent to 0.2 mg/mL of clindamycin from Topical Solution in the Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
Clindamycin Phosphate Topical Suspension

(Comment on this Monograph)id=m18305=Clindamycin Phosphate Topical Suspension=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Phosphate Topical Suspension contains the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of clindamycin (C18H33ClN2O5S). IDENTIFICATION The retention time of the clindamycin phosphate peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 13.6 mg/mL of monobasic potassium phosphate in water. Adjust with phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile and Solution A (9:31) System suitability solution: 0.6 mg/mL each of USP Clindamycin Phosphate RS and USP Clindamycin Hydrochloride RS, in the Mobile phase
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portions of methylene chloride, and allow the filter to airdry. Acceptance criteria: The IR absorption of the Sample exhibits maxima at the same wavelengths as that of a similar preparation of USP Clindamycin Phosphate RS. B. The retention time of the clindamycin phosphate peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 13.6 mg/mL of monobasic potassium phosphate in water. Adjust with phosphoric acid to a pH of 2.5. Solution B: 10.54 mg/mL of monobasic potassium phosphate in water. Adjust with phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile and Solution A (9:31) System suitability solution: 0.24 mg/mL of USP Clindamycin Phosphate RS and 6 g/mL of USP Clindamycin Hydrochloride RS, in Solution B Standard solution: 0.24 mg/mL of USP Clindamycin Phosphate RS, in Solution B Sample solution: Transfer one Vaginal Insert to a suitable 100-mL container. Add 40 mL of isooctane, and seal the container tightly with a teflon-lined septum and crimp cap. Shake vigorously for 15 min until all of the Vaginal Insert is dissolved. Add 40.0 mL of Solution B. Recap the container tightly, and shake vigorously for NLT 30 min, taking care to avoid leakage. Allow the layers to separate, and remove a volume of the lower aqueous layer sufficient to perform the following steps. Pass the aqueous solution through a filter having a 5-m or finer porosity, discarding the first 2 mL of the filtrate. Collect the remaining filtrate, and prepare a solution equivalent to 0.2 mg/mL of clindamycin with Solution B. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1.0 mL/min Injection size: 35 L System suitability Sample: System suitability solution and Standard solution Suitability requirements Resolution: NLT 2.0 between clindamycin phosphate and clindamycin hydrochloride for the System suitability solution, when calculated: Result = 1.177 [(t2 t1)/(wh1 + wh2)] = retention time of clindamycin hydrochloride (min) = retention time of clindamycin phosphate (min) t1 wh1 = width of the clindamycin phosphate peak at half-height (min) = width of the clindamycin hydrochloride peak at wh2 half-height (min) Relative standard deviation: NMT 2.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H33ClN2O5S equivalent in the portion of Vaginal Insert taken: t2 Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
Standard solution: 0.25 mg/mL of USP Clindamycin Phosphate RS, in the Mobile phase Sample solution: Using a suitable hypodermic needle and syringe, transfer the equivalent to 20 mg of clindamycin from Topical suspension to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clindamycin phosphate and clindamycin are about 1.0 and 1.5, respectively.] Suitability requirements Resolution: NLT 6.0 between the clindamycin phosphate and clindamycin peaks, System suitability solution Column efficiency: NLT 1700 theoretical plates, in the System suitability solution, when calculated: 5.545 (tr/Wh/2)2 Tailing factor: NMT 1.3, System suitability solution Relative standard deviation: NMT 2.5%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H33ClN2O5S in each mL of the Topical Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) = nominal concentration of clindamycin phosphate CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 4.56.5 MINIMUM FILL 755:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clindamycin Hydrochloride RS USP Clindamycin Phosphate RS
Clindamycin Phosphate Vaginal Inserts

(Comment on this Monograph)id=m18304=Clindamycin Phosphate Vaginal Inserts=Chl-Cy-Monos.pdf) DEFINITION Clindamycin Phosphate Vaginal Inserts contain the equivalent of NLT 90.0% and NMT 110.0% of the labeled amount of clindamycin (C18H33ClN2O5S). IDENTIFICATION A. INFRARED ABSORPTION 197M Sample: Transfer a Vaginal Insert into a suitable container, add 120 mL of methylene chloride, insert a stopper, and shake until the Vaginal Insert is completely dissolved. Using a vacuum, pass through a methylene chloride-compatible filter having a 0.45 m porosity. Rinse the filter with several
USP 32
= concentration of USP Clindamycin Phosphate RS in the Standard solution (mg/mL) = nominal concentration of clindamycin phosphate CU in the Sample solution (mg/mL) [NOTEFor the Assay value, use the average of the determinations obtained in the test for Uniformity of Dosage Units 905, Content Uniformity.] Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements CS
Official Monographs / Clioquinol 117

add 1.0 mL of bis(trimethylsilyl) acetamide and 1.0 mL of Internal standard solution, and attach the cap. Heat in a water bath at 50 for 15 min, and then cool to ambient temperature. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.83-m 2-mm glass column packed with 3% liquid phase G3 on 80- to 100-mesh support S1AB Temperatures Column: Initial temperature is 200 for a conditioning period of NLT 16 h (not connected to the detector) and is then reduced to 165. Injector: 170 Detector: 250 Carrier gas: Helium Flow rate: 30 mL/min Injection size: 1 L [NOTEHydrogen and air are introduced into the detector at rates of 25 mL and 500 mL/min, respectively.] System suitability Sample: Standard solution [NOTEThe relative retention times for clioquinol and pyrene are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 between the clioquinol and the internal standard peaks Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H5ClINO in the portion taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clioquinol to the internal standard peak from the Sample solution RS = peak response ratio of clioquinol to the internal standard peak from the Standard solution CS = concentration of USP Clioquinol RS in the Standard solution (mg/mL) = nominal concentration of clioquinol in the CU Sample solution (mg/mL) Acceptance criteria: 93.0%100.5% RU IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, at controlled room temperature, or in a cool place. USP REFERENCE STANDARDS 11 USP Clindamycin Hydrochloride RS USP Clindamycin Phosphate RS
Clioquinol
(Comment on this Monograph)id=m18315=Clioquinol=Chl-CyMonos.pdf)
C9H5ClINO 8-Quinolinol, 5-chloro-7-iodo-; 5-Chloro-7-iodo-8-quinolinol [130-26-7].
305.50
DEFINITION Clioquinol, dried over phosphorus pentoxide for 5 h, contains NLT 93.0% and NMT 100.5% of C9H5ClINO (the 5-chloro-7iodo-8-quinolinol isomer). IDENTIFICATION A. Prepare a Standard solution as directed for Standard stock solution in the Assay, except to use 1.0 mL of pyridine instead of the Internal standard solution, and chromatograph as directed in the Assay: the chromatogram of the Sample solution obtained in the Assay exhibits a peak for clioquinol, the retention time of which corresponds with that exhibited by the Standard solution. B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 267 nm Medium: 3 N hydrochloric acid Solution: 5 g/mL Acceptance criteria: Absorptivities calculated on the dried basis do not differ by more than 3.0%. C. Heat 100 mg with 5 mL of sulfuric acid: copious violet vapors of iodine are evolved. ASSAY PROCEDURE Internal standard solution: 2 mg/mL of pyrene in pyridine Standard stock solution: 3 mg/mL of USP Clioquinol RS in a mixture of pyridine and n-hexane (4:1) Standard solution: Transfer 1.0 mL of the Standard stock solution to a screw-capped glass vial fitted with a septum, add 1.0 mL of bis(trimethylsilyl) acetamide and 1.0 mL of Internal standard solution, attach the cap, and mix. Heat in a water bath at 50 for 15 min, and then cool to ambient temperature. Sample stock solution: 3 mg/mL of Clioquinol, previously dried, in a mixture of pyridine and n-hexane (4:1) Sample solution: Transfer 1.0 mL of the Sample stock solution to a screw-capped glass vial fitted with a septum,
NMT 0.5%
SPECIFIC TESTS LOSS ON DRYING 731: Dry it over phosphorus pentoxide for 5 h: it loses NMT 0.5% of its weight. FREE IODINE AND IODIDE: Shake 1.0 g with 20 mL of water for 30 s, allow to stand for 5 min, and filter. To 10 mL of the filtrate add 1 mL of 2 N sulfuric acid, then add 2 mL of chloroform, and shake: no violet color appears in the chloroform (free iodine). To the mixture add 5 mL of 2 N sulfuric acid and 1 mL of potassium dichromate TS, and shake for 15 s: the color in the chloroform layer is no deeper than that produced in a control test made in the following manner. Dilute 2.0 mL of potassium iodide solution (1 in 6000) with water to 10 mL, add 6 mL of 2 N sulfuric acid, 1 mL of potassium dichromate TS, and 2 mL of chloroform, and shake for 15 s (0.05% of iodide). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clioquinol RS
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Clioquinol / Official Monographs
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System suitability Sample: Standard solution [NOTEThe relative retention times for clioquinol and pyrene are 0.6 and 1.0, respectively. ] Suitability requirements Resolution: NLT 3 between the clioquinol and the internal standard peaks Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H5ClINO in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clioquinol to the internal standard peak from the Sample solution = peak response ratio of clioquinol to the internal RS standard peak from the Standard solution = concentration of USP Clioquinol RS in the CS Standard solution (mg/mL) = nominal concentration of clioquinol in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clioquinol RS
Clioquinol Cream
(Comment on this Monograph)id=m18317=Clioquinol Cream=Chl-Cy-Monos.pdf) DEFINITION Clioquinol Cream contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H5ClINO in a suitable cream base. IDENTIFICATION A. Prepare a Standard solution as directed for Standard stock solution in the Assay, except to use 1.0 mL of pyridine instead of the Internal standard solution, and chromatograph as directed in the Assay: the chromatogram of the Sample solution obtained in the Assay exhibits a peak for clioquinol, the retention time of which corresponds with that exhibited by the Standard solution. B. PROCEDURE Sample solution: Place an equivalent to 25 mg of clioquinol, from Cream, in a 100-mL volumetric flask, add 75 mL of dilute hydrochloric acid (1 in 4), and heat on a steam bath to melt the Cream, shaking vigorously to extract the clioquinol. Cool under running water, and add dilute hydrochloric acid (1 in 4) to volume. Filter through paper, and dilute 3 mL of the filtrate with dilute hydrochloric acid (1 in 4) to 100 mL. Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Clioquinol RS, concomitantly measured. ASSAY PROCEDURE Internal standard solution: 2 mg/mL of pyrene in pyridine Standard stock solution: 3 mg/mL of USP Clioquinol RS in a mixture of pyridine and n-hexane (4:1) Standard solution: Transfer 1.0 mL of the Standard stock solution to a screw-capped glass vial fitted with a septum, add 1.0 mL of bis(trimethylsilyl) acetamide and 1.0 mL of Internal standard solution, attach the cap, and mix. Heat in a water bath at 50 for 15 min, and then cool to ambient temperature. Sample stock solution: Transfer equivalent to 150 mg of clioquinol, from Cream, to a 60-mL separator. Place the separator on its side in a vacuum oven at a pressure of 10 mm of mercury at 45 for 4 h. Remove the separator from the oven, allow to cool, add 15 mL of a mixture of pyridine and n-hexane (4:1), insert a polytef stopper. Transfer the mixture to a 50-mL volumetric flask, and rinse the separator with two 15-mL portions of the same solvent, shaking each time for 30 s. Transfer both rinsings to the volumetric flask, and dilute with the same solvent to volume. Sample solution: Transfer 1.0 mL of the Sample stock solution to a screw-capped glass vial fitted with a septum, add 1.0 mL of bis(trimethylsilyl)acetamide and 1.0 mL of Internal standard solution, and attach the cap. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.83-m 2-mm glass column packed with 3% liquid phase G3 on 80- to 100-mesh support S1AB Temperature Column: Initial temperature is 200 for a conditioning period of NLT 16 h (not connected to the detector) and is then reduced to 165. Injector: 170 Detector: 250 Carrier gas: Helium Flow rate: 30 mL/min Injection size: 1 L [NOTEHydrogen and air are introduced into the detector at rates of 25 mL and 500 mL/min, respectively.]
Clioquinol Ointment
(Comment on this Monograph)id=m18321=Clioquinol Ointment=Chl-Cy-Monos.pdf) DEFINITION Clioquinol Ointment contains NLT 90.0% and NMT 110.0% of the labeled amount of C9H5ClINO in a suitable ointment base. IDENTIFICATION A. Prepare a Standard solution as directed for the Standard stock solution in the Assay, except to use 1.0 mL of pyridine instead of the Internal standard solution, and chromatograph as directed in the Assay: the chromatogram of the Sample solution obtained in the Assay exhibits a peak for clioquinol, the retention time of which corresponds with that exhibited by the Standard solution. B. PROCEDURE Sample solution: Place an equivalent to 25 mg of clioquinol, from Ointment, in a 100-mL volumetric flask, add 75 mL of dilute hydrochloric acid (1 in 4), and heat on a steam bath to melt the Ointment, shaking vigorously to extract the clioquinol. Cool under running water, and add dilute hydrochloric acid (1 in 4) to volume. Filter through paper, and dilute 3 mL of the filtrate with dilute hydrochloric acid (1 in 4) to 100 mL. Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Clioquinol RS, concomitantly measured. ASSAY PROCEDURE Internal standard solution: 2 mg/mL of pyrene in pyridine Standard stock solution: 3 mg/mL of USP Clioquinol RS in a mixture of pyridine and n-hexane (4:1) Standard solution: Transfer 1.0 mL of the Standard stock solution to a screw-capped glass vial fitted with a septum, add 1.0 mL of bis(trimethylsilyl) acetamide and 1.0 mL of Internal standard solution, and attach the cap. Heat in a
USP 32
water bath at 50 for 15 min, and then cool to ambient temperature. Sample solution: Transfer equivalent to 150 mg of clioquinol, from Ointment, to a 125-mL separator. Add 75 mL of n-hexane. Add 15 mL of dimethylformamide, and mix for 1 min. Allow the layers to separate, and transfer the lower layer to a 50-mL volumetric flask. Repeat the extraction with separate 15-mL and 10-mL portions of dimethylformamide, and transfer the lower layers to the 50mL volumetric flask. Dilute with dimethylformamide to volume. Transfer 1.0 mL of this solution to a screw-capped glass vial fitted with a septum, and evaporate at 60 under a stream of nitrogen to dryness. Add 1.0 mL of a mixture of pyridine and n-hexane (4:1) to the residue, add 1.0 mL of bis(trimethylsilyl) acetamide and 1.0 mL of Internal standard solution, and attach the cap. Heat in a water bath at 50 for 15 min, then cool to ambient temperature. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.83-m 2-mm glass column packed with 3% liquid phase G3 on 80- to 100-mesh support S1AB Temperature Column: Initial temperature is 200 for a conditioning period of NLT 16 h (not connected to the detector) and is then reduced to 165. Injector: 170 Detector: 250 Carrier gas: Helium Flow rate: 30 mL/min Injection size: 1 L [NOTEHydrogen and air are introduced into the detector at rates of 25 mL and 500 mL/min, respectively.] System suitability Sample: Standard solution [NOTEThe relative retention times for clioquinol and pyrene are 0.6 and 1.0, respectively] Suitability requirements Resolution: NLT 3 between the clioquinol and the internal standard peaks Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H5ClINO in the portion of Ointment taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clioquinol to the internal standard peak from the Sample solution = peak response ratio of clioquinol to the internal RS standard peak from the Standard solution = concentration of USP Clioquinol RS in the CS Standard solution (mg/mL) = nominal concentration of clioquinol in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clioquinol RS
Compound Clioquinol Topical Powder

(Comment on this Monograph)id=m18323=Compound Clioquinol Topical Powder=Chl-Cy-Monos.pdf) DEFINITION Compound Clioquinol Topical Powder contains NLT 22.5% and NMT 27.5% of C9H5ClINO.
Clioquinol Lactic Acid Zinc Stearate Lactose To make 250 g 25 g 200 g 525 g 1000 g
Mix the Lactic Acid with the Lactose, then add the Clioquinol and the Zinc Stearate.
IDENTIFICATION PROCEDURE Sample solution: Place equivalent to 30 mg of clioquinol from Topical Powder, in a glass-stoppered, 50-mL conical flask, add 20 mL of 1 N sulfuric acid, and shake for 5 min. Analysis: Transfer 5 mL of the filtrate to a glass-stoppered test tube, add 5 drops of potassium dichromate TS and 2 mL of chloroform, and shake well. Acceptance criteria: A red-violet color develops in the chloroform layer. ASSAY PROCEDURE Standard solution: 5 g/mL of USP Clioquinol RS in 3 N hydrochloric acid Sample stock solution: Transfer a quantity of Topical Powder, equivalent to 50 mg of clioquinol, to a 200-mL volumetric flask, add 100 mL of 3 N hydrochloric acid, and shake by mechanical means for 15 min. Dilute with 3 N hydrochloric acid to volume. Filter a portion of the solution. Sample solution: Dilute 4.0 mL of the filtrate with 3 N hydrochloric acid to 200.0 mL. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 267 nm Cell: 1 cm Blank: 3 N hydrochloric acid Analysis Samples: Standard solution and Sample solution Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C9H5ClINO in the portion of Topical Powder taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Clioquinol RS in the Standard solution (g/mL) = nominal concentration of clioquinol in the Sample solution (g/mL)
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22.5%27.5%
USP 32
Temperature Column: 165 Injector: 170 Detector: 250 Carrier gas: Dry helium Flow rate: 30 mL/min Injection size: 1 L System suitability Sample: Standard solution [NOTEThe relative retention times for clioquinol and pyrene are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the analyte and internal standard peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Record the chromatograms to obtain NLT 40% of maximum recorder response, and measure the peak response of each component. Calculate the percentage of C9H5ClINO in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = ratio of the peak responses of the clioquinol peak to the internal standard peak from the Sample solution = ratio of the peak responses of the clioquinol peak to the internal standard peak from the Standard solution = concentration of USP Clioquinol RS in the Standard solution (mg/mL) = nominal concentration of clioquinol in the Sample solution (mg/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Clioquinol RS
Clioquinol and Hydrocortisone Cream

(Comment on this Monograph)id=m18327=Clioquinol and Hydrocortisone Cream=Chl-Cy-Monos.pdf) DEFINITION Clioquinol and Hydrocortisone Cream contains NLT 90.0% and NMT 110.0% of the labeled amounts of clioquinol (C9H5ClINO) and of hydrocortisone (C21H30O5) in a suitable cream base. IDENTIFICATION A. The chromatogram of the Sample solution obtained as directed in the Assay for Clioquinol exhibits a peak for clioquinol, the retention time of which corresponds to that exhibited by the Standard solution. B. The chromatogram of the Sample solution obtained as directed in the Assay for Hydrocortisone exhibits a peak for hydrocortisone, the retention time of which corresponds to that exhibited by the Standard solution. ASSAY CLIOQUINOL Internal standard solution: 2 mg/mL of pyrene in pyridine Standard stock solution: 3 mg/mL of USP Clioquinol RS in a mixture of pyridine and n-hexane (4:1) Standard solution: Transfer 1.0 mL of the Standard stock solution, 1.0 mL of N,O-bis(trimethylsilyl)acetamide and 1.0 mL of Internal standard solution to a suitable screw-capped glass vial, fitted with a polytef-lined septum, and mix. Heat on a water bath at 50 for 15 min, and cool to room temperature. Sample solution: Transfer equivalent to 150 mg of clioquinol from Cream to a 60-mL separator. Place the separator on its side in a vacuum oven at 45 for 4 h. Remove the separator, cool to room temperature, and add 15.0 mL of a mixture of pyridine and hexane (4:1). Insert the stopper in the separator, and mix until the specimen is completely dispersed. Transfer the contents of the separator to a 50-mL volumetric flask, rinse the separator with two 15mL portions of a mixture of pyridine and hexane (4:1), collecting the rinsings in the volumetric flask, and dilute with the same solvent mixture to volume. Immediately transfer 1 mL of this solution to a dry, screw-capped glass vial, and evaporate with the aid of gentle heat and a stream of nitrogen to dryness. Dissolve the residue in 1.0 mL of a mixture of pyridine and hexane (4:1), add 1 mL each of N,O-bis(trimethylsilyl)acetamide and Internal standard solution to the screw-capped glass vial, fitted with a polyteflined septum, and mix. Heat on a water bath at 50 for 15 min, and cool to room temperature. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m column packed with 3% liquid phase G3 on 80- to 100-mesh support SlAB
HYDROCORTISONE Mobile phase: Acetonitrile, methanol and water (1:1:2.75) Standard stock solution: 1 mg/mL of USP Hydrocortisone RS in alcohol Standard solution: Standard stock solution and alcohol (1:9) Sample solution: Transfer equivalent to 10 mg of hydrocortisone from Cream to a 50-mL centrifuge tube. Add 30 mL of alcohol and heat on a steam bath just to boiling. Shake for 15 min and centrifuge. Transfer the supernatant extract to a 100-mL volumetric flask. Repeat the extraction with two 20-mL portions of alcohol, combining the extracts in the 100-mL volumetric flask. Add alcohol to volume, mix, and filter. System suitability stock solution: 0.5 mg/mL of methylparaben in alcohol System suitability solution: Mix the System suitability stock solution, Standard stock solution, and alcohol (1:10:89) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Pre-injection guard column: Packing L2 Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for methylparaben and hydrocortisone are 0.6 and 1.0, respectively.]
USP 32
Suitability requirements Resolution: NLT 2.0 between the hydrocortisone and methylparaben peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H30O5 in the portion of Cream taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Hydrocortisone RS in the Standard solution (g/mL) = nominal concentration of hydrocortisone in the CU Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clioquinol RS USP Hydrocortisone RS

50-mL volumetric flask, and dilute with dimethylformamide to volume. Transfer 1.0 mL of this solution to a suitable size screw-capped vial, and evaporate the solution with the aid of nitrogen at 60 to dryness. Dissolve the residue in 1.0 mL of a mixture of pyridine and hexane (4:1), and pipet 1.0 mL of N,O-bis(trimethylsilyl)acetamide and 1.0 mL of Internal standard solution into the glass vial, fitted with a polyteflined septum, and securely close. Heat the vial on a water bath at 50 for 15 min, and cool to room temperature Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m column packed with 3% liquid phase G3 on 80- to 100-mesh support SlAB Temperature Column: 165 Injector: 170 Detector: 250 Carrier gas: Dry helium Flow rate: 30 mL/min Injection size: 1 L System suitability Sample: Standard solution [NOTEThe relative retention times for clioquinol and pyrene are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 3.0 between the analyte and internal standard peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution. Record the chromatograms to obtain NLT 40% of maximum recorder response, and measure the peak response of each component. Calculate the percentage of C9H5ClINO taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = ratio of the peak responses of the clioquinol peak to the internal standard peak from the Sample solution = ratio of the peak responses of the clioquinol peak to the internal standard peak from the Standard solution = concentration of USP Clioquinol RS in the Standard solution (mg/mL) = nominal concentration of clioquinol in the Sample solution (mg/mL)
Clioquinol and Hydrocortisone Ointment

(Comment on this Monograph)id=m18332=Clioquinol and Hydrocortisone Ointment=Chl-Cy-Monos.pdf) DEFINITION Clioquinol and Hydrocortisone Ointment contains NLT 90.0% and NMT 110.0% of the labeled amounts of C9H5ClINO and of hydrocortisone C21H30O5 in a suitable ointment base. IDENTIFICATION The Sample solution obtained as directed in the Assay for clioquinol exhibits a peak for clioquinol, the retention time of which corresponds to that exhibited by the Standard solution. The Sample solution obtained as directed in the Assay for hydrocortisone exhibits a peak for hydrocortisone, the retention time of which corresponds to that exhibited by the Standard solution. ASSAY CLIOQUINOL Internal standard solution: 2 mg/mL of pyrene in pyridine Standard stock solution: 3 mg/mL of USP Clioquinol RS in a mixture of pyridine and n-hexane (4:1) Standard solution: Transfer 1.0 mL of the Standard stock solution, 1.0 mL of N,O-bis(trimethylsilyl)acetamide and 1.0 mL of Internal standard solution to a suitable screw-capped glass vial fitted with a polytef-lined septum, and mix. Heat on a water bath at 50 for 15 min, and cool to room temperature Sample solution: Transfer equivalent to 150 mg of clioquinol from Ointment to a 125-mL separator. Add 75 mL of n-hexane, insert the stopper in the separator, and mix until the specimen is completely dispersed. Extract with 25 mL of dimethylformamide, collecting the extract in a 50-mL volumetric flask. Repeat the extraction with two 10-mL portions of dimethylformamide, collecting the extracts in the
HYDROCORTISONE Mobile phase: Acetonitrile, methanol and water (1:1:2.75) Standard stock solution: 1 mg/mL of USP Hydrocortisone RS in alcohol Standard solution: Standard stock solution and alcohol (1:9) Sample solution: Transfer equivalent to 10 mg of hydrocortisone from Ointment to a 50-mL centrifuge tube. Add 30 mL of alcohol and heat on a steam bath just to boiling. Shake for 15 min, and centrifuge. Transfer the supernatant extract to a 100-mL volumetric flask. Repeat the extraction with two 20-mL portions of alcohol, combining the extracts in the 100-mL volumetric flask. Add alcohol to volume, mix, and filter. System suitability stock solution: 0.5 mg/mL of methylparaben in alcohol. System suitability solution: Mix the System suitability stock solution, Standard stock solution, and alcohol (1:10:89). Chromatographic system (See Chromatography 621, System Suitability.)
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USP 32
Standard solution: Dissolve a quantity of USP Clobetasol Propionate RS in methanol and Internal standard solution to obtain a final solution of 0.04 mg/mL of USP Clobetasol Propionate RS and 0.08 mg/mL of beclomethasone dipropionate. System suitability solution: 0.001 mg of USP Clobetasol Propionate Related Compound A RS and 0.1 mg of USP Clobetasol Propionate RS/mL of Mobile phase Sample solution: Transfer 4 mg of Clobetasol Propionate to a 100-mL volumetric flask, add 40.0 mL of Internal standard solution, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for clobetasol propionate related compound A and clobetasol propionate are 1.1 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.5 between clobetasol propionate and clobetasol propionate related compound A Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution [NOTEThe relative retention times for clobetasol propionate and beclomethasone dipropionate are 1.0 and 1.6, respectively.] Calculate the percentage of C23H32ClFO3 in the portion taken: Result = (RU/RS) (CS/CU) 100
Mode: LC Detector: UV 254 nm Guard column: Packing L2 Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for methylparaben and hydrocortisone are 0.6 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the hydrocortisone and methylparaben peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution. Calculate the percentage of C21H30O5 taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Hydrocortisone RS in the Standard solution (g/mL) = nominal concentration of hydrocortisone in the CU Sample solution (unit/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS MINIMUM FILL 755: Meets the requirements
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clioquinol RS USP Hydrocortisone RS
Clobetasol Propionate
(Comment on this Monograph)id=m18334=Clobetasol Propionate=Chl-Cy-Monos.pdf)
= ratio of the clobetasol propionate peak area to the internal standard peak area from the Sample solution = ratio of the clobetasol propionate peak area to RS the internal standard peak area from the Standard solution = concentration of USP Clobetasol Propionate RS CS in the Standard solution (mg/mL) = nomimal concentration of clobetasol propionate CU in the Sample solution (mg/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS 231: NMT 20 ppm Organic Impurities PROCEDURE Solution A, Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Sample solution: 0.1 mg/mL of Clobetasol Propionate in Mobile phase Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Clobetasol Proprionate taken: Result = (rU/rT) 100 rU rT = peak area for each impurity = sum of the areas of all of the peaks
RU
C25H32ClFO5 466.97 Pregna-1,4-diene-3,20-dione, 21-chloro-9-fluoro-11-hydroxy-16methyl-17-(1-oxopropoxy)-, (11,16)-; 21-Chloro-9-fluoro-11,17-dihydroxy-16-methylpregna-1,4diene-3,20-dione 17-propionate [25122-46-7; 25122-41-2]. DEFINITION Clobetasol Propionate contains NLT 97.0% and NMT 102.0% of C25H32ClFO5, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197M ASSAY PROCEDURE Solution A: 0.05 M monobasic sodium phosphate. Adjust with 85% phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile, methanol, and Solution A (19:4:17) Internal standard solution: 0.2 mg/mL of beclomethasone dipropionate in methanol
USP 32
Acceptance criteria: Any individual impurity: NMT 1.0% Total impurities: NMT 2.5% SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: Approximately 196 SPECIFIC ROTATION 781S: +98 to +104 at 20 Sample solution: 10 mg/mL in dioxane LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clobetasol Propionate RS USP Clobetasol Propionate Related Compound A RS
Official Monographs / Clobetasol 123

Mode: LC Detector: UV 240 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for clobetasol propionate, clobetasol propionate related compound A, and beclomethasone dipropionate are 1.0, 1.1, and 1.6, respectively.] Suitability requirements Resolution: NLT 1.5 between clobetasol propionate and clobetasol propionate related compound A Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C25H32ClFO5 in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 = ratio of the clobetasol propionate peak area to the internal standard peak area from the Sample solution RS = ratio of the clobetasol propionate peak area to the internal standard peak area from the Standard solution CS = concentration of USP Clobetasol Propionate RS in the Standard solution (mg/mL) CU = nominal concentration of clobetasol propionate in the Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% RU PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Salmonella species, and the total aerobic microbial count does not exceed 100 cfu/g. PH 791: 4.57.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight containers. Store at controlled room temperature. Do not refrigerate. USP REFERENCE STANDARDS 11 USP Clobetasol Propionate RS USP Clobetasol Propionate Related Compound A RS
Clobetasol Propionate Cream

(Comment on this Monograph)id=m18336s6=Clobetasol Propionate Cream=Chl-Cy-Monos.pdf) DEFINITION Clobetasol Propionate Cream is Clobetasol Propionate in a suitable cream base. It contains NLT 90.0% and NMT 115.0% of the labeled amount of C25H32ClFO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Transfer a quantity of Cream equivalent to 0.75 mg of clobetasol propionate to a 25-mL plasticstoppered centrifuge tube. Add 10 mL of methanol, and cap. Heat in a 60 water bath for 4 min, remove the tube from the bath, and shake vigorously. Repeat the heating and shaking. Cool to room temperature, add 3.5 mL of water, and mix. Centrifuge at 3500 rpm for 10 min. Transfer 5 mL of the supernatant to a 100-mL separator, add 1 g of sodium chloride and 10 mL of water, and mix. Extract with 5 mL of chloroform by shaking for 1 min, collect the lower layer, and evaporate with the aid of a stream of nitrogen to dryness. Dissolve the residue in 0.5 mL of chloroform. Standard solution: Same concentration of USP Clobetasol Propionate RS as the Sample solution Developing solvent system: Chloroform, acetone, and alcohol (20:2:1) ASSAY PROCEDURE Solution A: 0.05 M monobasic sodium phosphate. Adjust with 50% sodium hydroxide solution to a pH of 5.5. Mobile phase: Acetonitrile, methanol, and Solution A (19:4:17) Internal standard solution: 0.2 mg/mL of beclomethasone dipropionate in methanol Standard solution: Dissolve a quantity of USP Clobetasol Propionate RS in methanol and Internal standard solution to obtain a final solution of 0.04 mg/mL of USP Clobetasol Propionate RS and 0.08 mg/mL of beclomethasone dipropionate. System suitability solution: 0.001 mg/mL of USP Clobetasol Propionate Related Compound A RS and 0.1 mg/mL of USP Clobetasol Propionate RS in Mobile phase Sample solution: A quantity of Cream equivalent to 1.0 mg of clobetasol propionate in 10.0 mL of Internal standard solution and 15.0 mL of methanol. Shake vigorously to disperse the Cream, and centrifuge at about 3500 rpm for 10 min. Filter a portion of the supernatant through a 0.45m filter. Chromatographic system (See Chromatography 621, System Suitability.)
Clobetasol Propionate Ointment

(Comment on this Monograph)id=m18338=Clobetasol Propionate Ointment=Chl-Cy-Monos.pdf) DEFINITION Clobetasol Propionate Ointment is Clobetasol Propionate in a suitable ointment base. It contains NLT 90.0% and NMT 115.0% of the labeled amount of C25H32ClFO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: Same concentration of USP Clobetasol Propionate RS as the Sample solution, in chloroform Sample solution: Equivalent to 1.0 mg of clobetasol propionate from Ointment, in a 25-mL plastic-stoppered
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centrifuge tube. Add 10 mL of methanol, and cap. Heat in a 70 water bath for 4 min, remove the tube from the bath, and shake vigorously. Repeat the heating and shaking. Freeze the mixture in an ice bath for 5 min, and centrifuge at about 3500 rpm for 10 min. Transfer 5 mL of the supernatant to a suitable vial. Evaporate with the aid of a stream of nitrogen to dryness. Dissolve the residue in 1.0 mL of chloroform. Developing solvent system: Chloroform, acetone, and alcohol (20:2:1) ASSAY PROCEDURE Solution A: 0.05 M monobasic sodium phosphate. Adjust with 85% phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile, methanol, and Solution A (19:4:17) Internal standard solution: 0.2 mg/mL of beclomethasone dipropionate, in methanol Standard solution: 0.04 mg/mL of USP Clobetasol Propionate RS and 0.08 mg/mL of beclomethasone dipropionate, in Internal standard solution and methanol (2:3) System suitability solution: 0.001 mg/mL of USP Clobetasol Propionate Related Compound A RS and 0.1 mg/mL of USP Clobetasol Propionate RS, in Mobile phase Sample solution: Equivalent of 1.0 mg of clobetasol propionate from Ointment, in a 125-mL separator. Add 30 mL of hexane, 10.0 mL of Internal standard solution, and shake. Collect the lower layer in a 25-mL volumetric flask. Extract the hexane remaining in the separator with two 5mL portions of Mobile phase, and combine all of the extracts in the 25-mL volumetric flask. Dilute with Mobile phase to volume, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for clobetasol propionate and clobetasol propionate related compound A are 1.0 and 1.1, respectively.] Suitability requirements Resolution: NLT 1.5 between clobetasol propionate and clobetasol propionate related compound A Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Sample: Standard solution and Sample solution [NOTEThe relative retention times for clobetasol propionate and beclomethasone dipropionate are 1.0 and 1.6, respectively.] Calculate the percentage of C25H32ClFO5 in the portion of Ointment taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = ratio of the clobetasol propionate peak area to the internal standard peak area obtained from the Sample solution = ratio of the clobetasol propionate peak area to the internal standard peak area obtained from the Standard solution = concentration of USP Clobetasol Propionate RS in the Standard solution (mg/mL) = nominal concentration of clobetasol propionate in the Sample solution (mg/mL)
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Salmonella species; and the total aerobic microbial count does not exceed 100 cfu/g. MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight containers. Store at controlled room temperature (15 to 30). Do not refrigerate. USP REFERENCE STANDARDS 11 USP Clobetasol Propionate RS USP Clobetasol Propionate Related Compound A RS
Clobetasol Propionate Topical Solution

(Comment on this Monograph)id=m18339=Clobetasol Propionate Topical Solution=Chl-Cy-Monos.pdf) DEFINITION Clobetasol Propionate Topical Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of C25H32ClFO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: Same concentration of USP Clobetasol Propionate RS as the Sample solution, in chloroform Sample solution: Equivalent to 1.5 mg of clobetasol propionate from Topical Solution, in a 50-mL separator, add 5 mL of water, and extract with 5 mL of chloroform. Collect the lower layer through a cotton wool plug and evaporate with the aid of a stream of nitrogen to dryness. Dissolve the residue in 2 mL of chloroform. Developing solvent system: Chloroform, acetone, and alcohol (20:2:1) ASSAY PROCEDURE Solution A: 0.05 M monobasic sodium phosphate. Adjust with 85% phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile, methanol, and Solution A (19:4:17) Internal standard solution: 0.2 mg/mL of beclomethasone dipropionate, in methanol Standard solution: 0.04 mg/mL of USP Clobetasol Propionate RS and 0.08 mg/mL of beclomethasone dipropionate, in Internal standard solution and methanol (2:3) System suitability solution: 0.001 mg/mL of USP Clobetasol Propionate Related Compound A RS and 0.1 mg/mL of USP Clobetasol Propionate RS, in Mobile phase Sample solution: Equivalent to 1.0 mg of clobetasol propionate from Topical Solution, in a 25-mL volumetric flask Add 10.0 mL of Internal standard solution, dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 240 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for clobetasol propionate, clobetasol propionate related compound A,
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and beclomethasone dipropionate are about 1.0, 1.1, and 1.6, respectively.] Suitability requirements Resolution: NLT 1.5 between clobetasol propionate and clobetasol propionate related compound A Column efficiency: NLT 5000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Sample: Standard solution and Sample solution Calculate the percentage of C25H32ClFO5 in the portion of Topical Solution taken: Result = (RU/RS) (CS/CU) 100 = ratio of the clobetasol propionate peak area to the Internal standard peak area from the Sample solution = ratio of the clobetasol propionate peak area to RS the Internal standard peak area from the Standard solution = concentration of USP Clobetasol Propionate RS in CS the Standard solution (mg/mL) = nominal concentration of clobetasol propionate CU in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Salmonella species, and the total aerobic microbial count does not exceed 100 cfu/g. MINIMUM FILL 755: Meets the requirements PH 791: 4.5 6.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at controlled room temperature. Do not refrigerate. USP REFERENCE STANDARDS 11 USP Clobetasol Propionate RS USP Clobetasol Propionate Related Compound A RS RU
Official Monographs / Clocortolone 125

IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Spectral range: At 238 nm Medium: Methanol Solution: 15 g/mL Absorptivities: Calculated on the dried basis, do not differ by more than 3.0% ASSAY PROCEDURE Standard stock solution: 0.75 mg/mL of USP Clocortolone Pivalate RS, in chloroform Standard solution: 30 g/mL of USP Clocortolone Pivalate RS from Standard stock solution, in methanol Sample stock solution: 0.75 mg/mL of Clocortolone Pivalate, in chloroform Sample solution: 30 g/mL of Clocortolone Pivalate, from Sample stock solution in methanol Blank solution: 250 mg of isoniazid and 0.3 mL of hydrochloric acid, in 500 mL of methanol Analysis: Transfer 10.0-mL portions of the Standard solution and the Sample solution to separate glass-stoppered, 50-mL conical flasks, and evaporate on a steam bath to dryness. To each flask, and to a third flask to provide the blank, add 15.0 mL of Blank solution. Swirl the contents of the flasks to dissolve the residues. Insert the stoppers securely in the flasks, and place in a water bath at 60 for 2.5 h. Cool to room temperature. Spectrometric conditions Mode: Spectrophotometry Absorbance wavelength: 405 nm Cell: 1 cm Samples: Standard solution, Sample solution, and Blank solution Calculate the percentage of C27H36ClFO5 in the portion of Clocortolone Pivalate taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Clocortolone Pivalate RS in the Standard solution (g/mL) CU = nominal concentration of clocortolone pivalate in the Sample solution (g/mL) Acceptance criteria: 97.0%103.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Sample: 100 mg Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Sample solution: 4 mg/mL of Clocortolone Pivalate, in chloroform and methanol (1:1) Standard solution: 4 mg/mL of USP Clocortolone Pivalate RS, in chloroform and methanol (1:1) Blank: Chloroform and methanol (1:1) Application volume: 100 L Developing solvent system: Cyclohexane and ethyl acetate (2:1) AU AS CS
Clocortolone Pivalate
(Comment on this Monograph)id=m18340=Clocortolone Pivalate=Chl-Cy-Monos.pdf)
495.02 C27H36ClFO5 Pregna-1,4-diene-3,20-dione, 9-chloro-21-(2,2-dimethyl-1oxopropoxy)-6-fluoro-11-hydroxy-16-methyl-, (6,11,16)-; 9-Chloro-6-fluoro-11,21-dihydroxy-16-methylpregna-1,4diene-3,20-dione 21-pivalate [34097-16-0]. DEFINITION Clocortolone Pivalate contains NLT 97.0% and NMT 103.0% of C27H36ClFO5, calculated on the dried basis.
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Application volume: 20 L Developing solvent system: Cyclohexane and ethyl acetate (2:1) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Locate the spots on the plate by viewing under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot obtained from the Sample solution corresponds to that obtained from the Standard solution. ASSAY PROCEDURE Solution A: 250 mg of isoniazid and 0.3 mL of hydrochloric acid, in 500 mL of methanol Standard solution: 0.06 mg/mL of USP Clocortolone Pivalate RS, in methanol Sample solution: Equivalent to 3 mg of clocortolone pivalate by using a plastic syringe equipped with a suitable cannula, in a 50-mL volumetric flask. Add 25 mL of methanol, and warm the flask in a 60 water bath for 10 min, with occasional swirling, to disperse the Cream. Cool to room temperature, and dilute with methanol to volume. Allow any insoluble material to settle. Blank solution: 0.3 mL of hydrochloric acid diluted with methanol to 500 mL Analysis: Transfer 10.0 mL of the Standard solution to a 25mL volumetric flask. Transfer 10.0-mL portions of the Sample solution into two separate 25-mL volumetric flasks labeled Sample solution and Assay Blank, respectively. Evaporate the contents of the three flasks with the aid of a stream of air or nitrogen to dryness. Transfer 20.0 mL of Solution A to the flasks containing the Standard solution, the Sample solution, and a fourth 25-mL volumetric flask labeled Reagent Blank. Pipet 20 mL of Blank solution into the flask labeled Assay Blank. Insert the stoppers securely in the flasks, and place in a water bath at 60 for 2.5 h, occasionally swirling the contents of each flask. Cool the flasks to room temperature, dilute with the Blank solution to volume, and mix. Centrifuge the Sample solution at high speed for 10 min. Spectrometric conditions: Mode: Spectrophotometry Absorbance wavelength: 390 nm Cell: 1 cm Samples: Standard solution, Sample solution, Assay blank, Reagent blank, and Blank solution as the solvent blank Calculate the percentage of C27H36ClFO5 in the portion of Cream taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution, corrected for the Assay blank and the Reagent blank = absorbance of the Standard solution, corrected AS for the Reagent blank = concentration of USP Clocortolone Pivalate RS in CS the Standard solution (mg/mL) CU = nominal concentration of clocortolone pivalate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 5.07.0, in a 1-in-10 aqueous dispersion MINIMUM FILL 755: Meets the requirements PARTICLE SIZE DETERMINATION: Place a small portion of Cream on a microscope slide, apply a cover slide, press slightly, and examine under 40x objective magnification using a suitable microscope equipped with polarized light. Scan the complete slide preparation, and record the size of the largest crystal found in reference to a calibrated grid: no AU
Analysis Samples: Standard solution, Blank, and Sample solution Activate the plate at 105 for 30 min before use. Apply the solutions as streaks 2.5 cm from the bottom of the appropriate section of the plate, and dry the streaks with a gentle current of air. Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Air-dry the plate, and develop the chromatogram a second time using the same Developing solvent system. Air-dry the plate, and locate the principal band occupied by the Standard solution by viewing under UV light. Mark this band as well as corresponding bands in the Blank and Sample solution sections of the plate. Quantitatively remove the silica gel from each band, and transfer to separate glass-stoppered, 50-mL centrifuge tubes. Add 25.0 mL of methanol to each tube, shake for NLT 20 min, and centrifuge. Concomitantly determine the absorbances of the supernatants from the Sample solution and the Standard solution against the Blank. Spectrometric conditions Mode: Spectrophotometry Absorbance wavelength: 238 nm Calculate the percentage of chromatographic impurities in the Sample solution taken: Result = 100 [(AU/AS) (CS/CU) 100] = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Clocortolone Pivalate RS in the Standard solution (mg/mL) = nominal concentration of clocortolone pivalate CU in the Sample solution (mg/mL) Acceptance criteria Total impurities: NMT 3.0% SPECIFIC TESTS SPECIFIC ROTATION 781S: +125 to +135 Sample solution: 40 mg/mL, in chloroform LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 1.0% of its weight. COLOR AND CLARITY OF SOLUTION: A 1 in 100 solution in chloroform is clear and practically colorless. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clocortolone Pivalate RS AU AS CS
Clocortolone Pivalate Cream

(Comment on this Monograph)id=m18350=Clocortolone Pivalate Cream=Chl-Cy-Monos.pdf) DEFINITION Clocortolone Pivalate Cream contains NLT 90.0% and NMT 110.0% of C27H36ClFO5 in a suitable cream base. It may contain suitable preservatives. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 0.5 mg/mL of USP Clocortolone Pivalate RS, in chloroform Sample solution: Equivalent to 1 mg of clocortolone pivalate from Cream, in a suitable separator. Add 5 mL of water, and extract with 10 mL of chloroform. Evaporate the chloroform layer to dryness, and dissolve the residue in 2 mL of methanol.
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particle in the Cream is greater than 50 microns when measured in the longitudinal axis. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clocortolone Pivalate RS
Official Monographs / Clofazimine 127

Chromatographic plate: Immediately before use, expose the plate to ammonia vapors for 30 min by suspending the plate in a tank containing a shallow layer of approximately 25 mL of Solution A. [NOTEPrevent the plate from coming into contact with the liquid] Analysis Samples: Standard solutions and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and air-dry. Examine the plate under short-wavelength UV light, and compare the intensities of any secondary spots observed in the chromatogram of the Sample solution with those of the principal spots in the chromatograms of the Standard solutions. Acceptance criteria Individual impurities: NMT 1.0% Total impurities: NMT 2.0% SPECIFIC TESTS LOSS ON DRYING 731: 0.5% of its weight. Dry it at 105 for 3 h: it loses NMT
Clofazimine
(Comment on this Monograph)id=m18360=Clofazimine=ChlCy-Monos.pdf)
473.40 C27H22Cl2N4 2-Phenazinamine, N,5-bis(4-chlorophenyl)-3,5-dihydro-3-[(1methylethyl)imino]-; 3-(p-Chloroanilino)-10-(p-chlorophenyl)-2,10-dihydro-2(isopropylimino)phenazine [2030-63-9]. DEFINITION Clofazimine contains NLT 98.5% and NMT 101.5% of C27H22Cl2N4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197S: 5% solution in methylene chloride B. The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A, as obtained in the Procedure for Organic Impurities. ASSAY PROCEDURE Sample: 300 mg of Clofazimine Analysis: Dissolve the Sample in 5 mL of chloroform, with the aid of heat if necessary. Add 20 mL of acetone and 5 mL of glacial acetic acid, and titrate with 0.1 N perchloric acid VS in glacial acetic acid, using a glass electrode and a calomel electrode with a saturated solution of potassium chloride as the bridge fluid and agar gel as the bridge. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 47.34 mg of C27H22Cl2N4. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution A: 0.5 mg/mL of USP Clofazimine RS in methylene chloride Standard solution B: 0.25 mg/mL of USP Clofazimine RS from Standard solution A in methylene chloride Standard solution C: 0.1 mg/mL of USP Clofazimine RS from Standard solution A in methylene chloride Sample solution: 50 mg/mL of Clofazimine in methylene chloride Solution A: Ammonium hydroxide and water (1:99) [NOTEPrepare this solution fresh daily.] Application volume: 5 L Developing solvent system: Methylene chloride and npropyl alcohol (10:1)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers at room temperature. USP REFERENCE STANDARDS 11 USP Clofazimine RS
Clofazimine Capsules
(Comment on this Monograph)id=m18365=Clofazimine Capsules=Chl-Cy-Monos.pdf) DEFINITION Clofazimine Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of C27H22Cl2N4. IDENTIFICATION The RF value of the principal spot observed in the chromatogram of the Sample solution corresponds to that of the Standard solution, obtained as directed in the Procedure for Organic Impurities. The UV absorption spectrum of the Sample solution, prepared as directed in the Assay, exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Clofazimine RS, concomitantly measured. ASSAY PROCEDURE Solution A: Hydrochloric acid and methanol (1:99) Reference solution: Methylene chloride and Solution A (1:9) Standard stock solution: 0.075 mg/mL of USP Clofazimine RS in methylene chloride Standard solution: Standard stock solution and Solution A (1:9) Sample stock solution: Remove as completely as possible the contents of NLT 20 Capsules with the aid of methylene chloride. Dissolve in methylene chloride, filter the solution through a pledget of cotton, and prepare 0.075 mg/mL solution with methylene chloride.
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rotation of the blade. Observe the Capsules, and record the time taken for each capsule shell to rupture. Tolerances: The requirements are met if all the Capsules tested rupture in NMT 15 min. If 1 or 2 of the Capsules rupture in more than 15 but NMT 30 min, repeat the test on 12 additional Capsules. NMT 2 of the total of 18 Capsules tested rupture in more than 15 but NMT 30 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clofazimine RS
Sample solution: Sample stock solution and Solution A (1:9) Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 491 nm Blank: Reference solution Analysis: Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C27H22Cl2N4 taken: Result = (AU/As) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Clofazimine RS in the Standard solution (mg/mL) CU = nominal concentration of clofazimine in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% AU AS CS IMPURITIES Organic Impurities PROCEDURE Standard solution A: 0.5 mg/mL of USP Clofazimine RS in methylene chloride Standard solution B: 0.1 mg/mL of USP Clofazimine RS from Standard solution A in methylene chloride Standard solution C: 0.04 mg/mL of USP Clofazimine RS from Standard solution A in methylene chloride Sample solution: To a portion of Capsule contents equivalent to 500 mg of clofazimine, add 25 mL of methylene chloride and 25 mL of 0.1 N sodium hydroxide, and sonicate for 30 min. Withdraw the methylene chloride layer, and filter through anhydrous sodium sulfate. Solution A: Ammonium hydroxide and water (1:99) [NOTEPrepare this solution fresh daily.] Chromatographic system Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Methylene chloride and npropyl alcohol (10:1) Chromatographic plate: Immediately before use, expose the plate to ammonia vapors for 30 min by suspending the plate in a tank containing a shallow layer of approximately 25 mL of Solution A. [NOTEPrevent the plate from coming into contact with the liquid.] Analysis Samples: Standard solutions and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and air-dry. Examine the plate under short-wavelength UV light, and compare the intensities of any secondary spots observed from the Sample solution with those of the principal spots from the Standard solutions. Acceptance criteria Individual impurities: NMT 1.0% Total impurities: NMT 2.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 500 mL Apparatus 2: 50 rpm Time: 15 min Analysis: Place 1 Capsule in each vessel, and allow the Capsule to sink to the bottom of the vessel before starting
Clofibrate
(Comment on this Monograph)id=m18400=Clofibrate=Chl-CyMonos.pdf)
C12H15ClO3 242.70 Propanoic acid, 2-(4-chlorophenoxy)-2-methyl-, ethyl ester; Ethyl 2-(p-chlorophenoxy)-2-methylpropionate [637-07-0]. DEFINITION Clofibrate contains NLT 97.0% and NMT 103.0% of C12H15ClO3, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197F B. ULTRAVIOLET ABSORPTION 197U Medium: Methanol Solution: 20 g/mL ASSAY PROCEDURE Ion-exchange resin: To a beaker containing 75 mL of 1 N sodium hydroxide add 3 g of a 50- to 100-mesh strongly basic styrene-divinylbenzene anion-exchange resin, and allow the mixture to stand for 15 min, with occasional stirring. Wash the resin with water until the last washing is neutral to litmus paper, then wash with three 50-mL portions of methanol. Ion-exchange column: Place a plug of glass wool in the base of a 1- 15-cm ion-exchange tube, and transfer to the tube a sufficient amount of Ion-exchange resin, slurried in methanol, to produce a column bed height of from 6 cm to 8 cm. Standard solution: 20 g/mL of USP Clofibrate RS, in methanol Sample solution: 2 mg/mL of Clofibrate, in methanol. Transfer 10.0 mL of this solution to the Ion-exchange column, and collect the eluate in a 100-mL volumetric flask. Rinse the column with 25 mL of methanol, collect the rinsing in the volumetric flask, and dilute with methanol to volume. Dilute with methanol to 20 g/mL of clofibrate.
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Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 226 nm Cell: 1 cm Blank: Methanol Analysis Samples: Standard solution and Sample solution Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance (226 nm). Calculate the percentage of C12H15ClO3 in the portion taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Clofibrate RS in the Standard solution (g/mL) = nominal concentration of clofibrate in the CU Sample solution (g/mL) Acceptance criteria: 97.0%103.0% IMPURITIES Organic Impurities PROCEDURE 1 Standard solution: 0.1 mg/mL of USP Clofibrate RS and 0.03 mg/mL of p-chlorophenol in chloroform. To 10.0 mL of this solution add 5.0 L of tributyrin. Sample solution: To 10.0 mL of Clofibrate add 5.0 L of tributyrin. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.53-mm 15-m column to the internal walls of which is bonded a 1.5-m film of phase G1 Temperature Column
Time (min) 0 113 1322 Temperature 120 120180 (5/min) 180
Official Monographs / Clofibrate 129

= ratio of the areas of the clofibrate peak to that of the tributyrin peak from the Standard solution = concentration of USP Clofibrate RS in the CS Standard solution (mg/mL) Acceptance criteria Any individual impurity (other than p-chlorophenol): NMT 0.01% Total of all such impurities: NMT 0.12% PROCEDURE 2: LIMIT OF p-CHLOROPHENOL Analysis: Use the chromatograms obtained in Procedure 1. Calculate the percentage of p-chlorophenol in the portion of Clofibrate taken: Result = (RU/RS) CS 0.1 = ratio of the areas of the p-chlorophenol and tributyrin peaks from the Sample solution RS = ratio of the areas of the p-chlorophenol and tributyrin peaks from the Standard solution CS = concentration of p-chlorophenol in the Standard solution (mg/mL) Acceptance criteria: NMT 0.003% of p-chlorophenol SPECIFIC TESTS REFRACTIVE INDEX 831: 1.5001.505, at 20 ACIDITY Analysis: Mix 10.0 g with 100 mL of neutralized alcohol, add 3 drops of phenolphthalein TS, and titrate with 0.10 N sodium hydroxide. Acceptance criteria: NMT 0.90 mL is required for neutralization. WATER DETERMINATION, Method I 921: NMT 0.2% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Clofibrate RS RU RS
AU AS CS
Clofibrate Capsules
(Comment on this Monograph)id=m18410=Clofibrate Capsules=Chl-Cy-Monos.pdf) DEFINITION Clofibrate Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of C12H15ClO3. IDENTIFICATION INFRARED ABSORPTION 197F ASSAY PROCEDURE Ion-exchange resin: To a beaker containing 75 mL of 1 N sodium hydroxide add 3 g of a 50- to 100-mesh strongly basic styrene-divinylbenzene anion-exchange resin, and allow the mixture to stand for 15 min, with occasional stirring. Wash the resin with water until the last washing is neutral to litmus paper, then wash with three 50-mL portions of methanol. Ion-exchange column: Place a plug of glass wool in the base of a 1- 15-cm ion-exchange tube, and transfer to the tube a sufficient amount of Ion-exchange resin, slurried in methanol, to produce a column bed height of from 6 cm to 8 cm. Standard solution: 20 g/mL of USP Clofibrate RS, in methanol Sample solution: Weigh accurately NLT 20 Capsules in a tared weighing bottle. With a sharp blade, carefully open the Capsules, without loss of shell material, and transfer the
Injector: 210 Detector: 220 Carrier gas: Helium Flow rate: 2 mL/min Injection size: 1 L Injection type: Split injector Split ratio: 20:1 System suitability Sample: Standard solution Suitability requirements [NOTERelative retention times: The relative retention times for p-chlorophenol, clofibrate, and tributyrin are about 0.2, 0.55, and 1.0, respectively.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity, other than pchlorophenol, in the portion taken: Result = (RU/RS) CS 0.1 RU = ratio of the areas of each individual impurity peak (other than the p-chlorophenol peak) to that of the tributyrin peak from the Sample solution
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contents to a 100-mL beaker. Remove any liquid from the emptied capsules by washing with several small portions of ether. Discard the washings, and allow the capsules to dry in a jet of dry air until the odor of ether no longer is perceptible. Weigh the empty capsules in the tared weighing bottle, and calculate the average net weight/capsule. Transfer an accurately weighed amount of capsule contents, equivalent to 200 mg of clofibrate, to a 100-mL volumetric flask, add methanol to volume, and mix. Transfer 10.0 mL of this solution to the Ion-exchange column, and collect the eluate in a 100-mL volumetric flask. Rinse the column with 25 mL of methanol, collect the rinsings in the volumetric flask, dilute with methanol to volume, and mix. Dilute with methanol to 20 g/mL of clofibrate. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 226 nm Cell: 1 cm Blank: Methanol Analysis: Determine the absorbances of the Standard solution and the Sample solution. Calculate the percent of C12H15ClO3 in the Capsules taken: Result = (AU/AS) (CS/CU) 100 AU AS CS = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Clofibrate RS in the Standard solution (g/mL) = nominal concentration of clofibrate in the Sample CU solution (g/mL) Acceptance criteria: 90.0%110.0%
Clomiphene Citrate
(Comment on this Monograph)id=m18460=Clomiphene Citrate=Chl-Cy-Monos.pdf)
598.08 C26H28ClNO C6H8O7 Ethanamine, 2-[4-(2-chloro-1,2-diphenylethenyl)phenoxy]-N,Ndiethyl-, 2-hydroxy-1,2,3-propanetricarboxylate (1:1); 2-[p-(2-Chloro-1,2-diphenylvinyl)phenoxy]triethylamine citrate (1:1) [50-41-9]. DEFINITION Clomiphene Citrate contains NLT 98.0% and NMT 102.0% of a mixture of the (E)- and (Z)-geometric isomers of C26H28ClNO C6H8O7, calculated on the anhydrous basis. It contains NLT 30.0% and NMT 50.0% of the Z-isomer, [(Z)-2-[4-(2chloro-1,2-diphenylethenyl)phenoxy]-N,N-diethylethanamine 2-hydroxy-1,2,3-propanetricarboxylate (1:1). IDENTIFICATION A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181 B. ULTRAVIOLET ABSORPTION 197U Medium: 0.1 N hydrochloric acid Solution: 20 g/mL C. IDENTIFICATION TESTSGENERAL 191, Citrate ASSAY PROCEDURE Mobile phase: Methanol, water, and triethylamine (55:45:0.3). Adjust with phosphoric acid to a pH of 2.5. System suitability solution: 2 g/mL of USP Clomiphene Related Compound A RS and 50 g/mL of USP Clomiphene Citrate RS, in Mobile phase [NOTEUse actinic glassware for all solutions.] Standard solution: 50 g/mL of USP Clomiphene Citrate RS, in Mobile phase Sample solution: 50 g/mL of Clomiphene Citrate, in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 233 nm Column: 4.6-mm 25-cm; packing L26 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times are about 0.9 for clomiphene related compound A, 1.0 for (Z)-isomer, and 1.2 for (E)-isomer.] Suitability requirements Resolution: NLT 1.0 between clomiphene related compound A and (Z)-isomer, and NLT 1.5 between (Z)isomer and (E)-isomer, System suitability solution Column efficiency: NLT 2000 theoretical plates for the (E)isomer, Standard solution Tailing factor: NMT 3.0 for the (E)-isomer, Standard solution Relative standard deviation: NMT 2.0% for both (E)- and (Z)-isomers, Standard solution
PERFORMANCE TESTS DISSOLUTION 711 Medium: Sodium lauryl sulfate solution (5 in 100); 1000 mL Apparatus 2: 100 rpm Time: 180 min Determine the amount of C12H15ClO3 dissolved by employing the following method. Mobile phase: Methanol and water (4:1) Standard solution: Dissolve 20 mg of USP Clofibrate RS in 20 mL of methanol and dilute with water to 50 mL. Dilute with methanol to 80 g/mL of USP Clofibrate RS. Sample solution: Dilute a 5.0-mL portion of the sample to 25.0 mL, using methanol. Allow to stand for 5 min, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 226 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the amount of C12H15ClO3 dissolved. Tolerances: NLT 80% (Q) of the labeled amount of C12H15ClO3 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Clofibrate RS
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of C26H28ClNO C6H8O7 in the portion of Clomiphene Citrate taken: Result = ((rUE + rUZ)/(rSE + rSZ)) (CS/CU) 100 = peak response for the (E)-isomer from the Sample solution = peak response for the (Z)-isomer from the rUZ Sample solution = peak response for the (E)-isomer from the rSE Standard solution = peak response for the (Z)-isomer from the rSZ Standard solution CS = concentration of USP Clomiphene Citrate RS in the Standard solution (g/mL) = nominal concentration of clomiphene citrate in CU the Sample solution (g/mL) Acceptance criteria: 98.0%102.0% OTHER COMPONENTS Content of (Z)-isomer Mobile phase, Standard solution, System suitability solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Analysis Samples: Sample solution Calculate the percentage of (Z)-isomer in the portion of Clomiphene Citrate taken: Result = (rZ/rT) 100 = peak response for the (Z)-isomer from the Sample solution rT = sum of all the peak responses of the Sample solution Acceptance criteria: 30.0%50.0% rZ IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities RELATED COMPOUNDS Mobile phase, Sample solution, and System suitability solution: Proceed as directed in the Assay Standard solution: 1 g/mL of USP Clomiphene Citrate RS, in Mobile phase Chromatographic system Mode: LC Detector: UV 290 nm Column: 4.6-mm 25-cm; packing L26 Flow rate: 1.0 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times are about 0.9 for clomiphene related compound A, 1.0 for the (Z)-isomer, and 1.2 for the (E)-isomer.] Suitability requirements Resolution: NLT 1.0 between clomiphene related compound A and the (Z)-isomer, and NLT 1.5 between the (Z)-isomer and the (E)-isomer, System suitability solution Column efficiency: NLT 2000 theoretical plates for the (E)-isomer, Standard solution Tailing factor: NMT 3.0 for the (E)-isomer, Standard solution Relative standard deviation: NMT 2.0% from both the (E)- and (Z)-isomers, Standard solution rUE
Official Monographs / Clomiphene 131

Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Clomiphene Citrate taken: Result = (rU/rT) 100 rU = peak response for each impurity = sum of all the peak responses rT Acceptance criteria Clomiphene related compound A: NMT 2.0% Other individual impurity: NMT 0.5% Total impurities: NMT 2.0% SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 1.0%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clomiphene Citrate RS USP Clomiphene Related Compound A RS
Clomiphene Citrate Tablets

(Comment on this Monograph)id=m18490=Clomiphene Citrate Tablets=Chl-Cy-Monos.pdf) DEFINITION Clomiphene Citrate Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of clomiphene citrate (C26H28ClNO C6H8O7). IDENTIFICATION IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Analysis: Place a portion of finely powdered Tablets equivalent to 30 mg of clomiphene citrate in a centrifuge tube containing 30 mL solution of methanol and 0.1 N hydrochloric acid (1:2). Insert the stopper, and place the tube in a water bath at 37 for 15 min. Shake occasionally. Centrifuge, and place the clear supernatant in a separator. Extract with one 40-mL and two 25-mL portions of hexanes, and discard the extract. Render the aqueous solution alkaline with 1 N sodium hydroxide, and extract the precipitated base with one 50-mL and two 25-mL portions of hexanes. Wash the combined extracts with two portions of water. Dry the extract with anhydrous sodium sulfate, and remove the hexanes by evaporation under reduced pressure. Add 1.0 mL of carbon disulfide to the residue, and dissolve. Determine the absorption spectra of the Sample solution and a Standard solution of USP Clomiphene Citrate RS, similarly prepared, as directed under IdentificationOrganic Nitrogenous Bases 181. ASSAY PROCEDURE Mobile phase: Methanol, water, and triethylamine (55:45:0.3). Adjust with phosphoric acid to a pH of 2.5. System suitability solution: 0.002 mg/mL of USP Clomiphene Related Compound A RS and 0.05 mg/mL of USP Clomiphene Citrate RS in Mobile phase [NOTEUse actinic glassware for Standard solution and Sample solution.] Standard solution: 50 g/mL of USP Clomiphene Citrate RS, in Mobile phase Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer 50 mg of clomiphene citrate equivalent to a 100mL volumetric flask. Add about 50 mL of Mobile phase, and stir using a magnetic bar for 30 min. Remove the magnetic
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bar from the flask, dilute with Mobile phase to volume, and filter. Dilute the resulting solution with Mobile phase. Filter, and discard the first 10 mL. [NOTEThis solution is stable for NLT 24 h.] Chromatographic system (See under Chromatography 621 System Suitability.) Mode: LC Detector: UV 233 nm Column: 4.6-mm 25-cm; packing L26 Flow rate: 1.0 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for clomiphene related compound A, (Z)-isomer, and (E)-isomer are 0.9, 1.0, and 1.2, respectively.] Suitability requirements Resolution: NLT 1.0 between clomiphene related compound A and (Z)-isomer, and NLT 1.5 between (Z)isomer and (E)-isomer, System suitability solution Column efficiency: NLT 2000 theoretical plates for the (E)-isomer, Standard solution Tailing factor: NMT 3.0 for the (E)-isomer, Standard solution Relative standard deviation: NMT 2.0% for both (E)and (Z)-isomers, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C26H28ClNO C6H8O7 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clomiphene Citrate RS in the Standard solution (g/mL) CU = nominal concentration of clomiphene citrate in the Sample solution (g/mL) Acceptance criteria: 93.0%107.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 1: 100 rpm Time: 30 min Detector: UV 232 nm Sample solutions: Sample per Dissolution 711. Dilute with 0.1 N hydrochloric acid to a concentration that is similar to the Standard solution. Standard solution: USP Clomiphene Citrate RS in 0.1 N hydrochloric acid Tolerances: NLT 75% (Q) of the labeled amount of C26H28ClNO C6H8O7 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, protected from light. USP REFERENCE STANDARDS 11 USP Clomiphene Citrate RS USP Clomiphene Related Compound A RS
Clomipramine Hydrochloride
(Comment on this Monograph)id=m18510=Clomipramine Hydrochloride=Chl-Cy-Monos.pdf)
351.31 C19H23ClN2 HCl 5H-Dibenz[b,f]azepine-5-propanamine, 3-chloro-10,11-dihydroN,N-dimethyl-, monohydrochloride; 3-Chloro-5-[3-(dimethylamino)propyl]-10,11-dihydro-5H-dibenz [b,f]azepine monohydrochloride [17321-77-6]. DEFINITION Clomipramine Hydrochloride contains NLT 98.0% and NMT 102.0% of C19H23ClN2 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Medium: 0.1 N hydrochloric acid Solution: 100 g/mL Acceptance criteria: Absorptivities do not differ by more than 1.0% at the wavelength of maximum absorbance, calculated on the dried basis. ASSAY PROCEDURE Solution A: Dissolve 5.5 g of sodium 1-heptanesulfonate in 50.0 mL of water, and dilute to 100.0 mL with glacial acetic acid. Mobile phase: Transfer 20.0 mL of Solution A and 2.0 mL of triethylamine to a 500-mL volumetric flask, and dilute with water to volume. Transfer this solution to a 1-L volumetric flask, adjust with phosphoric acid to a pH of 3.2 0.1, dilute with acetonitrile to volume, mix, filter, and degas. System suitability solution: 0.07 mg/mL of USP Desipramine Hydrochloride RS and 0.10 mg/mL of USP Imipramine Hydrochloride RS in methanol Standard solution: 0.32 mg/mL of USP Clomipramine Hydrochloride in methanol Sample solution: 0.32 mg/mL of Clomipramine Hydrochloride RS in methanol Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for desipramine and imipramine are 0.85 and 1.0, respectively.]
USP 32
Suitability requirements Resolution: NLT 0.5 between desipramine and imipramine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H23ClN2 HCl in the portion of Clomipramine Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clomipramine Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of Clomipramine Hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 100 ppm Organic Impurities PROCEDURE 1 Solution A and System suitability solution: Proceed as directed in the Assay. Mobile phase: Transfer 20.0 mL of Solution A, 2.0 mL of triethylamine, and 500 mL of water to a suitable container, mix, adjust with phosphoric acid to a pH of 3.2 0.1, and dilute with water to 625 mL. Transfer to a 1-L volumetric flask, and dilute with acetonitrile to volume. Sample solution: 2 mg/mL of Clomipramine Hydrochloride in methanol Chromatographic system: Using an Injection size of 5 L, proceed as directed in the Assay. Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Clomipramine Hydrochloride taken: Result = (rU/rT) 100 = peak response for each impurity rU rT = sum of all the peak responses Acceptance criteria: NMT 0.5% for any individual impurity PROCEDURE 2 Solution A, Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Sample solution: Prepare as directed for Procedure 1. Analysis Samples: Sample solution Calculate the percentage of each impurity in the portion of Clomipramine Hydrochloride taken by the formula: Result = (rU/rS) 100 = peak response for each impurity rU = sum of the responses of all the peaks rS Acceptance criteria: Individual impurity: NMT 0.5% Total impurities: NMT 2.0% [NOTEThe results for Total impurities include individual results fromProcedure 1 and Procedure 2.] SPECIFIC TESTS PH 791: 3.55.0, in 100 mg/mL solution LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 1.0% of its weight.
Official Monographs / Clomipramine 133

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clomipramine Hydrochloride RS USP Desipramine Hydrochloride RS USP Imipramine Hydrochloride RS
Clomipramine Hydrochloride Capsules

(Comment on this Monograph)id=m18515=Clomipramine Hydrochloride Capsules=Chl-Cy-Monos.pdf) DEFINITION Clomipramine Hydrochloride Capsules contain NLT 90.0% and NMT 110.0% of clomipramine hydrochloride (C19H23ClN2 HCl). IDENTIFICATION INFRARED ABSORPTION 197K Sample: Transfer the equivalent to 125 mg of clomipramine hydrochloride from Capsules in 25 mL of chloroform, stir for 5 min, and filter. Evaporate on a steam bath to a volume of 5 mL, chill in an ice bath, add ethyl ether, and stir until crystals form. Filter, and dry at 100 for 1 h. ASSAY PROCEDURE Solution A: Dissolve 5.5 g of sodium 1-heptanesulfonate in 50.0 mL of water, and dilute to 100.0 mL with glacial acetic acid. Mobile phase: Transfer 20.0 mL of Solution A and 2.0 mL of triethylamine to a 500-mL volumetric flask, and dilute with water to volume. Transfer this solution to a 1-L volumetric flask, adjust with phosphoric acid to a pH of 3.2 0.1, dilute with acetonitrile to volume, mix, filter, and degas. System suitability solution: 0.07 mg/mL of USP Desipramine Hydrochloride RS and 0.10 mg/mL of USP Imipramine Hydrochloride RS in methanol Standard solution: 0.32 mg/mL of USP Clomipramine Hydrochloride RS in methanol Sample solution: Remove as completely as possible the contents of NLT 20 Capsules. Transfer the equivalent to 160 mg of Clomipramine hydrochloride to a 200-mL volumetric flask, add 130 mL of methanol, shake by mechanical means for 1 h, and dilute with methanol to volume. Transfer 10 mL of this solution to a 25-mL volumetric flask, dilute with methanol to volume, mix, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution [NOTEThe relative retention times for desipramine and imipramine are 0.85 and 1.0, respectively.] Suitability requirements Resolution: NLT 0.5 between desipramine and imipramine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H23ClN2 HCl in the portion of Capsules taken: Result = (rU/rS) (CS/CU) (100/L)
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rU
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= peak response for Clomipramine from the Sample solution rS = peak response from the Standard solution CS = concentration of USP Clomipramine Hydrochloride RS in the Standard solution (mg/mL) = concentration of the Sample solution CU (capsule/mL) L = label claim (mg/capsule) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 30 min Analytical wavelengh: UV 252 nm Sample solutions: Sample per Dissolution 711 Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Clomipramine Hydrochloride RS in Medium Tolerances: NLT 80% (Q) of the labeled amount of C19H23ClN2 HCl UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Content uniformity Standard solution: 30 g/mL of USP Clomipramine Hydrochloride RS in methanol Sample solution: Quantitatively transfer the contents of 1 Capsule to a 100-mL volumetric flask with the aid of methanol. Add 75 mL of methanol, shake by mechanical means for 1 h, and dilute with methanol to volume. Prepare a solution having a nominal concentration of 30 g/mL with methanol. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 252 nm Cell: 1 cm Blank: Methanol Analysis: Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C19H23ClN2 HCl in the portion of Capsules taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = = = = absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution(g/mL)
Clonazepam
(Comment on this Monograph)id=m18540=Clonazepam=ChlCy-Monos.pdf)
C15H10ClN3O3 315.71 2H-1,4-Benzodiazepin-2-one, 5-(2-chlorophenyl)-1,3-dihydro-7nitro-; 5-(o-Chlorophenyl)-1,3-dihydro-7-nitro-2H-1,4-benzodiazepin-2one [1622-61-3]. DEFINITION Clonazepam contains NLT 98.0% and NMT 102.0% of C15H10ClN3O3, calculated on the dried basis. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Diluent: Methanol, tetrahydrofuran, and water (52:13:60) Solution A: 6.6 g/L of anhydrous dibasic ammonium phosphate in water Adjust with 1 N phosphoric acid or 1 N sodium hydroxide to a pH of 8.0. Mobile phase: Methanol, tetrahydrofuran, and Solution A (52:13:60) System suitability solution: 0.04 mg/mL each of USP Clonazepam Related Compound A RS, USP Clonazepam Related Compound B RS, and USP Clonazepam RS in Diluent Standard solution: 0.1 mg/mL of USP Clonazepam RS in Diluent Sample solution: 0.1 mg/mL of Clonazepam in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times are 2.2 for clonazepam related compound A, 2.5 for clonazepam related compound B, and 1.0 for clonazepam.] Suitability requirements Resolution: NLT 2.0 between clonazepam related compound A and clonazepam related compound B, System suitability solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 2.0%, Standard solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clomipramine Hydrochloride RS USP Desimipramaine hydrochloride RS USP Imipramine Hydrochloride RS
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H10ClN3O3 taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clonazepam RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities LIMIT OF CLONAZEPAM RELATED COMPOUND C Standard solution: 50 g/mL of USP Clonazepam Related Compound C RS in acetone Sample solution: 25 mg/mL of Clonazepam in acetone Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Developing solvent system: Acetone and n-heptane (3:2) Spray reagent 1: 2 M sulfuric acid Spray reagent 2: 0.01 M sodium nitrite Spray reagent 3: 9 mM ammonium sulfamate Spray reagent 4: N-(1-naphthyl)ethylenediamine dihydrochloride TS Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. After air-drying the plate, heavily spray the plate with Spray Reagent 1, and dry at 105 for 15 min. Successively spray the plate with Spray reagents 2, 3, and 4, and dry the plate with a current of air. Compare the intensities of any secondary spots observed in the Sample solution with that of the principal spot of the Standard solution. Acceptance criteria: No secondary spot of the Sample solution is larger or more intense than the principal spot of the Standard solution (0.2%). PROCEDURE Buffer, Mobile phase, Diluent, System suitability solution, Standard solution, and Chromatographic system: Proceed as directed in the Assay. Sample solution: Use the Sample solution in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Clonazepam taken: Result = F rU1/(rU2 + F rU1) 100 rU1 rU2 = peak response for each impurity of the Sample solution = peak response for clonazepam in the Sample solution rU rS CS
Official Monographs / Clonazepam 135

F = relative response factor Acceptance criteria Individual impurities: See Impurity Table 1. Sum of all other impurities: NMT 0.3%
Impurity Table 1 Relative Response Factor 1.84 0.94 1.00 Acceptance Criteria, NMT (%) 0.1 0.1 0.2
Impurity Name Clonazepam related compound A Clonazepam related compound B Any other impurity
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 237240 LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in containers at room temperature. USP REFERENCE STANDARDS 11 USP Clonazepam RS USP Clonazepam Related Compound USP Clonazepam Related Compound USP Clonazepam Related Compound tight, light-resistant
A RS B RS C RS
Clonazepam Oral Suspension

(Comment on this Monograph)id=m1384=Clonazepam Oral Suspension=Chl-Cy-Monos.pdf) DEFINITION Clonazepam Oral Suspension contains NLT 90.0% and NMT 110.0% of the labeled amount of clonazepam (C15H10ClN3O3). Prepare Clonazepam Oral Suspension 0.1 mg/mL as follows. (See Pharmaceutical CompoundingNonsterile Solutions 795.)
Clonazepam Vehicle: a mixture of Vehicle for Oral Solution, (regular or sugar-free), NF and Vehicle for Oral Suspension, NF (1:1), To make 10 mg A sufficient quantity
100 mL
If using Tablets, comminute the Tablets into a fine powder in a suitable mortar, or add Clonazepam powder to the mortar. Add approximately 10 mL of the Vehicle, and mix to a uniform paste. Add the Vehicle in small portions almost to volume, and mix thoroughly after each addition. Transfer the contents of the mortar, stepwise and quantitatively, to a calibrated bottle. Add enough Vehicle to bring to final volume, and mix well.
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Clonazepam / Official Monographs
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nitrogen to dryness. Wash the residue with three 10-mL portions of solvent hexane. Acceptance criteria: The IR absorption spectrum of a potassium bromide dispersion of the Sample solution exhibits maxima only at the same wavelengths as that of a similar preparation of USP Clonazepam RS. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Methanol, tetrahydrofuran, and water (52:13:60) Solution A: 6.6 g/L of anhydrous dibasic ammonium phosphate in water Adjust with 1 N phosphoric acid or 1 N sodium hydroxide to a pH of 8.0. Mobile phase: Methanol, tetrahydrofuran, and Solution A (52:13:60) System suitability solution: 0.04 mg/mL each of USP Clonazepam Related Compound A RS, USP Clonazepam Related Compound B RS, and USP Clonazepam RS in Diluent Standard solution: 0.1 mg/mL of USP Clonazepam RS in Diluent Sample solution: Finely powder NLT 10 Tablets. Transfer an amount of powder equivalent to 10 mg of clonazepam to a 100-mL volumetric flask, and dissolve, with sonication, in 75 mL of Diluent. Cool to room temperature, dilute with Diluent to volume, mix, and filter, discarding the first few mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clonazepam related compound A, clonazepam related compound B, and clonazepam are 2.2, 2.5, and 1.0, respectively.] Suitability requirements Resolution: NLT 2.0 between clonazepam related compound A and clonazepam related compound B, System suitability solution Tailing factor: NMT 1, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H10ClN3O3 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response for clonazepam from the Sample solution = peak response for clonazepam from the Standard rS solution = concentration of USP Clonazepam RS in the CS Standard solution (mg/mL) = nominal concentration of clonazepam in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION 711: Determine the amount of Clonazepam dissolved, using the following method.
ASSAY PROCEDURE Mobile phase: Methanol, acetonitrile, and water (4:3:3) Standard solution: 25 g/mL of USP Clonazepam RS in acetonitrile Sample solution: Agitate the container of Oral Suspension for 30 min on a rotating mixer, remove a 5-mL sample, and store in a clear glass vial at 70 until analyzed. At the time of analysis, remove the sample from the freezer, allow it to reach room temperature, and mix with a vortex mixer for 30 s. Pipet 2.5 mL of the Sample solution into a 10-mL volumetric flask, and dilute with acetonitrile to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 10-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe retention time of Clonazepam is about 7 min.] Suitability requirements Relative standard deviation: NMT 1.8% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H10ClN3O3 in the volume of Oral Suspension: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clonazepam RS in the Standard solution (g/mL) = nominal concentration of clonazepam in the CU Sample solution (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 3.64.6 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at controlled room temperature, or in a cold place. LABELING: Label it to state that it is to be well shaken before use, and to state the beyond-use date. BEYOND-USE DATE: 60 days after the day on which it was compounded. USP REFERENCE STANDARDS 11 USP Clonazepam RS rU rS CS
Clonazepam Tablets
(Comment on this Monograph)id=m18570=Clonazepam Tablets=Chl-Cy-Monos.pdf) DEFINITION Clonazepam Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of clonazepam (C15H10ClN3O3). IDENTIFICATION A. INFRARED ABSORPTION Sample solution: Place an amount of finely powdered Tablets, equivalent to 10 mg of clonazepam in a 125-mL separator. Add 25 mL of water, shake for 2 min, and extract with two 40-mL portions of chloroform. Pass the extracts through anhydrous sodium sulfate, combine them, and evaporate at room temperature with the aid of a stream of
USP 32
Medium: Degassed water; 900 mL Apparatus 2: 75 rpm Time: 45 min Mobile phase: Methanol, acetonitrile, and water (3:3:4) Sample solution: Sample per Dissolution 711 Standard stock solution: 0.05 mg/mL of USP Clonazepam RS in methanol Dilute a portion of the Standard stock solution with Medium to obtain a known concentration similar to the expected concentration in the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.0 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0 Analysis Samples: Standard solution and Sample solution Calculate the percentage of C15H10ClN3O3 dissolved by comparison of the peak responses of the Standard solution and the Sample solution. Tolerances: NLT 75% (Q) of the labeled amount of C15H10ClN3O3 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements IMPURITIES Organic Impurities PROCEDURE Solution A, Mobile phase, Diluent, System suitability solution, Standard solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Sample solution: Use the Sample solution in the Assay. Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Tablets taken by the formula: Result = rU1/(rU2 + F rU1) F 100 = peak response for each impurity from the Sample solution rU2 = peak response of clonazepam from the Sample solution F = relative response factor (see Impurity Table 1) Acceptance criteria Individual impurities: See Impurity Table 1. Sum of all other impurities: NMT 0.5% rU1
Official Monographs / Clonidine 137

USP REFERENCE STANDARDS 11 USP Clonazepam RS USP Clonazepam Related Compound A RS USP Clonazepam Related Compound B RS
Clonidine
(Comment on this Monograph)id=m18600=Clonidine=Chl-CyMonos.pdf)
230.09 C9H9Cl2N3 Benzenamine, 2,6-dichloro-N-2-imidazolidinylidene-; 2-[(2,6-Dichlorophenyl)imino]imidazolidine [4205-90-7]. DEFINITION Clonidine contains NLT 99.0% and NMT 101.0% of C9H9Cl2N3, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Medium: 0.01 N hydrochloric acid Solution: 0.3 mg/mL Acceptance criteria: Absorptivities are about 2.1 and 1.8 for the maxima at 271 and 279, respectively, calculated on the dried basis ASSAY PROCEDURE Sample solution: 190 mg of Clonidine in 100 mL of glacial acetic acid Analysis: Titrate with 0.1 N perchloric acid VS, using a silver-silver chloride glass combination electrode with liquid junction. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 23.01 mg of C9H9Cl2N3. Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Ignite at 500 25 to constant weight. HEAVY METALS, Method II 231: NMT 10 ppm CHROMATOGRAPHIC PURITY 1 M phosphoric acid: 115 mg/mL of phosphoric acid in water Solution A: 4 mg/mL of monobasic potassium phosphate in water Adjust with 1 M phosphoric acid to a pH of 4.0. Solution B: Acetonitrile and Solution A (3:1) Blank: Solution A Mobile phase: Use variable mixtures of Solution A and Solution B as directed for Chromatographic system Acetylclonidine solution: Dissolve about 1 mg of USP Clonidine Related Compound A RS in 1 mL of acetonitrile, and dilute with Solution A to 20 mL. System suitability solution: 0.86 mg/mL of USP Clonidine RS and 0.86 g/mL of acetylclonidine from Acetylclonidine solution in Solution A Standard solution: 8.6 g/mL of USP Clonidine RS in Solution A
Name Clonazepam related compound A Clonazepam related compound B Unknown impurity Any other impurity
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, at room temperature.
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Clonidine / Official Monographs
USP 32
Sample solution: 0.86 mg/mL of Clonidine in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 3.0 mm 15 cm; m packing L56 Temperature: 40 Flow rate: 1.5 mL/min Program: See the table below.
Time (min) 0 15 15.1 20 Solution A (%) 90 30 90 90 Solution B (%) 10 70 10 10
Clonidine Hydrochloride
(Comment on this Monograph)id=m18620=Clonidine Hydrochloride=Chl-Cy-Monos.pdf)
C9H9Cl2N3 HCl 266.55 Benzenamine, 2,6-dichloro-N-2-imidazolidinylidene-, monohydrochloride; 2-[(2,6-Dichlorophenyl)imino]imidazolidine monohydrochloride [4205-91-8]. DEFINITION Clonidine Hydrochloride contains NLT 98.5% and NMT 101.0% of C9H9Cl2N3 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: At 272 nm Medium: 0.01 N hydrochloric acid Solution: 330 g/mL Acceptance criteria: Absorptivities, calculated on the dried basis do not differ by more than 3.0%. C. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements ASSAY PROCEDURE Sample solution: To 200 mg of Clonidine Hydrochloride in 80 mL of glacial acetic acid, add 15 mL of mercuric acetate TS. Analysis: Titrate with 0.1 N perchloric acid VS, using a glass electrode and a sleeve-type calomel electrode containing 0.1 N lithium perchlorate in glacial acetic acid (see Titrimetry 541). Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 26.66 mg of C9H9Cl2N3 HCl. Acceptance criteria: 98.5%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution A: 100 mg/mL of USP Clonidine Hydrochloride RS in methanol Standard solution B: 100 g/mL of USP Clonidine Hydrochloride RS from Standard solution A in methanol Sample solution: 100 mg/mL in methanol Spray reagent: Starch-potassium iodide TS Application volume: 2 L Developing solvent system: Toluene, dioxane, dehydrated alcohol, and ammonium hydroxide (10:8:2:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in a freshly prepared Developing solvent system, until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the chamber, allow the solvent to
Injection size: 5 L System suitability Sample: System suitability solution and Standard solution Suitability requirements Resolution: NLT 5 between clonidine and acetylclonidine, System suitability solution Tailing factor: NMT 2.5 for clonidine, System suitability solution Relative standard deviation: NMT 5.0%, Standard solution Analysis Samples: Blank, Standard solution, and Sample solution Calculate the percentage of each impurity in the Clonidine taken: Result = (rU/rS) (CS/CU) 100 = peak response for each impurity from the Sample solution rS = peak response of clonidine from the Standard solution CS = concentration of the Standard solution (g/mL) CU = concentration of the Sample solution (g/mL) Acceptance criteria: Individual impurity: NMT 0.1% Total impurities: NMT 0.2% rU SPECIFIC TESTS LOSS ON DRYING 731: Dry it under vacuum at 60 to constant weight: it loses NMT 0.5% of its weight. APPEARANCE OF SOLUTION Color of solution Sample solution: 100 mg/mL in methanol Standard solution: 4.8 g/mL of potassium chromate in water Acceptance criteria: The color of 10 mL of Sample solution is not more intense than 10 mL of a Standard solution, when compared in matched color-comparison tubes (see Spectrophotometry and Light-Scattering 851, Visual Comparison). Turbidity: The Sample solution prepared as directed under Color of solution has no more turbidity than 10 mL of methanol, when compared in matched color-comparison tubes (see Spectrophotometry and Light-Scattering 851, Visual Comparison). ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at room temperature. USP REFERENCE STANDARDS 11 USP Clonidine RS USP Clonidine Related Compound A RS
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evaporate, and dry it at 100 for 1 h. Dip the plate into a dipping chamber filled to three-fourths of its height with sodium hypochlorite solution, diluted to contain 0.5% available chlorine, dry in a fume hood with a current of air for 1 h, and spray with Spray reagent: the RF value of the principal spot from the Sample solution corresponds to that of Standard solution A. Any other spot obtained from the Sample solution does not exceed, in size or intensity, the principal spot obtained from Standard solution B. Acceptance criteria Individual impurities: NMT 0.1% Total impurities: NMT 0.2% SPECIFIC TESTS PH 791: 3.55.5, in 50 mg/mL solution LOSS ON DRYING 731: Dry it at 105 to constant weight: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25; excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Clonidine Hydrochloride RS

ASSAY PROCEDURE Solution A: 2.2 mg/mL of sodium 1-octanesulfonate Mobile phase: Methanol, Solution A, and phosphoric acid (500:500:1) Adjust with 1 N sodium hydroxide to a pH of 3.0, and filter through a 0.45-m or finer porosity filter. 2,6-Dichloroaniline stock solution: 12 g/mL of 2,6dichloroaniline in Mobile phase Standard stock solution: 100 g/mL of USP Clonidine Hydrochloride RS in Mobile phase System suitability solution: Transfer 2.0 mL of Standard stock solution, and 20.0 mL of 2,6-Dichloroaniline stock solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Standard solution: 1 g/mL of clonidine hydrochloride from the Standard stock solution in Mobile phase Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer the equivalent to 0.1 mg of clonidine hydrochloride from powder to a 100-mL volumetric flask. Add about 60 mL of Mobile phase, shake by mechanical means for 15 to 30 min, dilute with Mobile phase to volume, and mix. Centrifuge a portion of this solution to obtain a clear solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L7, deactivated for basic compounds Flow rate: 1.5 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for clonidine and 2,6dichloroaniline are about 0.5 and 1.0, respectively.] Suitability requirements Column efficiency: NLT 3500 theoretical plates from the clonidine peak, System suitability solution Tailing factor: NMT 1.5 for the clonidine peak, System suitability solution Relative standard deviation: NMT 2.0% for the clonidine peak, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H9Cl2N3 HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (g/mL) nominal concentration of clonidine hydrochloride in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS CU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 500 mL Apparatus 2: 50 rpm Time: 30 min Analysis: Using Medium instead of Mobile phase to prepare the Standard stock solution and the Standard solution, determine the amount of C9H9Cl2N3 HCl dissolved, employing the Procedure in the Assay, and making any necessary modifications. = = = =
Clonidine Hydrochloride Tablets

(Comment on this Monograph)id=m18650=Clonidine Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Clonidine Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C9H9Cl2N3 HCl. IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 10 mg/mL of USP Clonidine Hydrochloride RS in methanol Sample solution: Transfer an equivalent to 1 mg of clonidine hydrochloride, from a quantity of finely powdered Tablets, to a separator containing 30 mL of water, and 5 mL of 1 N sodium hydroxide. Swirl gently to dissolve the sample specimen, and extract with 20 mL of chloroform. Allow the layers to separate, and filter the chloroform extract. Evaporate the filtrate to dryness, and dissolve the residue in 0.1 mL of methanol. Application volume: 2 L Developing solvent system: Methanol and ammonium hydroxide (200:3) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Position the plate in a chromatographic chamber, and develop the chromatograms in the Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Examine the plate under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution.
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150J g/mL of USP Clonidine Hydrochloride RS and about 150 g/mL of USP Chlorthalidone RS Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer the equivalent to 15 mg of chlorthalidone from powder to a 100-mL volumetric flask, add 10 mL of methanol, and sonicate for 5 min. Add 40 mL of 0.1% monobasic ammonium phosphate solution, sonicate until the solution is free from agglomerates, allow to cool to ambient temperature, dilute with 0.1% monobasic ammonium phosphate solution to volume, mix, and centrifuge. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 10-cm; packing L7 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for clonidine hydrochloride and chlorthalidone are about 0.2 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 is between the clonidine hydrochloride and the chlorthalidone peaks. Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H9Cl2N3 HCl in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 = peak response of clonidine hydrochloride from the Sample solution = peak response of clonidine hydrochloride from RS the Standard solution = concentration of USP Clonidine Hydrochloride RS CS in the Standard solution (g/mL) = nominal concentration of clonidine CU hydrochloride in the Sample solution (g/mL) Calculate the percentage of C14H11ClN2O4S in the portion of Tablets taken by the same formula, changing the terms to refer to chlorthalidone. Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 100 rpm Time: 60 min Analysis: Pipet 20 mL of a centrifuged portion of the solution under test into a 25-mL volumetric flask, and dilute with 0.5% monobasic ammonium phosphate solution to volume. Use the resulting solution as the Sample solution. Determine the amounts of C14H11ClN2O4S and C9H9Cl2N3 HCl dissolved, employing the procedure set forth in the Assay, making any necessary volumetric adjustments. Tolerances: NLT 50% (Q) of the labeled amount of C14H11ClN2O4S and NLT 80% (Q) of the labeled amount of C9H9Cl2N3 HCl UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity with respect to both clonidine hydrochloride and chlorthalidone ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Chlorthalidone RS USP Clonidine Hydrochloride RS
Tolerances: NLT 75% (Q) of the labeled amount of C9H9Cl2N3 HCl UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clonidine Hydrochloride RS
Clonidine Hydrochloride and Chlorthalidone Tablets

(Comment on this Monograph)id=m18660=Clonidine Hydrochloride and Chlorthalidone Tablets=Chl-Cy-Monos.pdf) DEFINITION Clonidine Hydrochloride and Chlorthalidone Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of chlorthalidone (C14H11ClN2O4S) and NLT 90.0% and NMT 110.0% of the labeled amount of clonidine hydrochloride (C9H9Cl2N3 HCl). IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: Transfer an amount of powdered Tablets, equivalent to 3 mg of clonidine hydrochloride, to a beaker, add 30 mL of water, stir for 5 min, and filter, using a medium-porosity, sintered-glass funnel. Transfer the filtrate to a separator, add 5 mL of 0.1 N sodium hydroxide, and extract with 20 mL of chloroform, collecting the chloroform extract in a separator. Extract the chloroform phase with 15 mL of 0.01 N hydrochloric acid, collecting the acid extract in a beaker. Remove any residual chloroform from the acid extract by heating on a steam bath. Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Clonidine Hydrochloride RS, concomitantly measured. B. INFRARED ABSORPTION Sample: Transfer 10 powdered Tablets to a 50-mL beaker, add 10 mL of methanol, boil on a steam bath for 5 min, and filter. Add 20 mL of water to the filtrate, and boil on a steam bath for 5 min under a current of air. Cool, with stirring, in ice until crystals form, filter the crystals, and dry at 105 for 1 h. Acceptance criteria: The IR absorption spectrum of a mineral oil dispersion of the Sample exhibits maxima only at the same wavelengths as that of a similar preparation of USP Chlorthalidone RS. C. The retention times of the chlorthalidone and clonidine hydrochloride peaks in the chromatogram of the Sample solution correspond to those of the Standard solution as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 800 mg of monobasic ammonium phosphate in 800 mL of water, and add 100 mL of methanol and 100 mL of acetonitrile. Standard solution: 1500J g/mL of USP Clonidine Hydrochloride RS in 0.1% monobasic ammonium phosphate solution, J being the ratio of the labeled amount, in mg, of clonidine hydrochloride to the labeled amount, in mg, of chlorthalidone per Tablet (Solution P). Transfer 15 mg of USP Chlorthalidone RS to a 100-mL volumetric flask, dissolve in 10 mL of methanol, add 25 mL of 0.1% monobasic ammonium phosphate solution and 10.0 mL of Solution P. Dilute with 0.1% monobasic ammonium phosphate solution to volume, and mix to obtain a solution having known concentrations of about
USP 32

For systems containing about 2.5 mg of clonidine For systems containing about 5.0 mg of clonidine For systems containing about 7.5 mg of clonidine 7.0 mL 14.0 mL 21.0 mL
Clonidine Transdermal System

(Comment on this Monograph)id=m18610=Clonidine Transdermal System=Chl-Cy-Monos.pdf) DEFINITION Clonidine Transdermal System contains NLT 80.0% and NMT 120.0% of the labeled amount of C9H9Cl2N3. [NOTEThroughout the following procedures, avoid the use of tetrahydrofuran stabilized with butylated hydroxytoluene (BHT). In the presence of peroxides, BHT may react with clonidine, producing impurity peaks.] IDENTIFICATION A. INFRARED ABSORPTION 197K Solution A: Dissolve 242.28 mg/mL of tris(hydroxymethyl)aminomethane in water. Adjust to a pH of 9.2, using dilute hydrochloric acid. Sample: Carefully peel the release liner from each Transdermal System, and place a number of Transdermal Systems equivalent to 25 mg of clonidine into a 50-mL screw-capped centrifuge tube. Add 5 mL of chloroform, and mix on a vortex mixer for 5 min. Allow to stand for 30 min, and mix intermittently on a vortex mixer. Transfer the chloroform solution to another 50-mL centrifuge tube, and wash the residue with an additional 3 mL of chloroform, combining the extracts. Add 2 mL of 0.5 N hydrochloric acid to the extract, mix on a vortex mixer for 1 min, and centrifuge at about 1000 rpm for 4 min. Remove and discard the bottom chloroform layer. Extract the aqueous layer with 4 mL of chloroform. Centrifuge at 1000 rpm for an additional 5 min, and again discard the bottom chloroform layer. Add 5 mL of Solution A and 3 mL of methylene chloride. Mix on a vortex mixer for 1 min. Centrifuge at 1000 rpm for 4 min. Transfer the bottom methylene chloride layer into a 100-mL beaker, and dry the methylene chloride with anhydrous sodium sulfate (about 1/4 liquid height). Decant and evaporate to dryness with a stream of nitrogen. Dry at 105 for 30 min, and allow to cool in a desiccator. Analysis: Determine the IR spectrum of the Sample solution and USP Clonidine Hydrochloride RS in the wavelength region of 3500 to 600 cm1. B. The retention time of the major peak from the Sample solution corresponds to that of the Standard solution as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Dissolve 4 mL of triethylamine in 1.6 L of water, and adjust with phosphoric acid to a pH of 3.0. Add 2.4 L of acetonitrile and stir the solution for 30 min. Diluent: Tetrahydrofuran and methanol (1:1) System suitability solution: 250 g/mL of USP Clonidine RS and 10 g/mL of USP Clonidine Related Compound B RS in Diluent Standard stock solution: 1 mg/mL of USP Clonidine RS in tetrahydrofuran Standard solutions: Prepare a minimum of four Standard solutions in Diluent that bracket the expected clonidine concentration in the sample. The standard concentrations should be within the range of 50300 g/mL. [NOTEStandard solutions are stable for up to 2 days if stored at 4.] Sample solution: Remove each Transdermal System from its package, discard the release liner from each system, and transfer into a 50-mL centrifuge tube with a Teflon-lined screw cap. Add the appropriate volume of tetrahydrofuran as listed in the table below.
Mix vigorously on a vortex mixer until the systems are washed down and fully submerged in the tetrahydrofuran. Let the systems soak in tetrahydrofuran for about 5 min, and mix on a vortex mixer until the systems are completely delaminated. Allow the systems to remain submerged for an additional 60 min, mixing on a vortex mixer every 30 min. Add methanol in a volume equal to the volume of tetrahydrofuran, and mix vigorously on a vortex mixer. The solution turns milky. Centrifuge for 10 min at 2000 rpm. Use the supernatant as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm and 242 nm [NOTEThe detector is programmed initially to 242 nm and 210 nm after the elution of the clonidine peak but prior to the elution of the clonidine related compound B.] Column: 4.6-mm 15-cm; packing L10 Flow rate: 1 mL/min Injection size: 25 L System suitability Sample: System suitability solution [NOTEThe relative retention times are 1.0 and 1.5 for clonidine and clonidine related compound B, respectively.] Suitability requirements Resolution: NLT 2.0 between clonidine and clonidine related compound B Capacity factor, k: NLT 0.6 for clonidine Tailing factor: NMT 3.0 for both clonidine and clonidine related compound B Relative standard deviation: NMT 2.0% from the clonidine peak area Analysis Samples: At least three Standard solutions that will bracket the expected sample concentration range and the Sample solution Calculate the peak response ratios of the analyte, and plot the results. Determine the linear regression equation of the standards by the mean-square method, and record the linear regression equation and the correlation coefficient; it should be NLT 0.995. Calculate the amount, in mg, of C9H9Cl2N3 in the Transdermal System taken: Result = CV/1000 = concentration of clonidine from the linear regression analysis (g/mL) V = volume of the Sample solution in mL/sample system Acceptance criteria: 80.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements DRUG RELEASE 724: Medium: 0.001 M phosphoric acid; 80 mL for systems containing 5 mg or less of clonidine, 200 mL for systems containing more than 5 mg of clonidine Times: 8, 24, 96, and 168 h Apparatus 7: Proceed as directed in the chapter, using the transdermal system holder-angled disk (see Figure 4a). The C
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T = time (h) A = area of the Transdermal System (cm2) Tolerances: The release rate of C9H9Cl2N3 from the Transdermal System, expressed as g/h/cm2 at the times specified, conforms to Drug Release 724, Acceptance Table 1.
Time (h) 08 824 2496 96168 Time for Sampling (h) 8 24 96 168 Release Rate (g/h/cm2) 7.516.0 1.54.6 1.54.6 1.53.3
appropriate size of the holder, 1.42 or 1.98 inches, should be chosen based on the size of the system to prevent overhang. Use 100-mL beakers for Medium volumes of 80 mL and 300 mL beakers for Medium volumes of 200 mL. Gently press the transdermal system to a dry, smooth, square piece of cellulose membrane, or equivalent, with the adhesive side against the membrane. Attach the membrane/system to a suitable inert sample holder with a Viton O-ring, or equivalent, so that the backing of the system is adjacent to and centered on the bottom of the sample holder. Trim the excess cellulose membrane with scissors. Suspend each sample holder from the arm of a reciprocating shaker so that each system is continuously immersed in a beaker containing the specified volume of Medium. The filled beakers are weighed and preequilibrated to 32.0 0.3 prior to immersing the test sample. Agitate the sample in an up-down motion at a frequency of 30 cycle min with an amplitude of 2.0 0.1 cm. The Medium must be added daily to the beakers during each interval to maintain sample immersion. At the end of each time interval, transfer the test sample to a fresh beaker containing the appropriate volume of Medium, weighed and pre-equilibrated to 32.0 0.3. Determine the amount of C9H9Cl2N3 released by employing the following method: Mobile phase: Use a 0.1% solution of triethylamine in a mixture of methanol and water (3:7), and adjust with phosphoric acid to a pH of 6.0 0.2. System suitability solution: 10 g/mL of USP Clonidine RS in 0.001 M phosphoric acid Standard solutions: Prepare a minimum of four standard solutions of USP Clonidine RS in 0.001 M phosphoric acid having known concentrations of clonidine similar to those of the Sample solutions. Sample solutions: At the end of each release interval, allow the beakers to cool to room temperature and make up for evaporative Medium losses by adding Medium to obtain the original weight, then mix. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 25 L System suitability Sample: System suitability solution Suitability requirements Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Capacity factor: NLT 0.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solutions and Sample solutions Construct a standard curve of concentration (g per mL) of clonidine in the Standard solutions versus peak area by linear regression analysis. The correlation coefficient is NLT 0.995. Calculate the release rate of clonidine: Result = CV/TA C V
A
IMPURITIES Organic Impurities RELATED COMPOUNDS Mobile phase, Diluent, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Standard stock solution: 1 mg/mL of USP Clonidine Related Compound B RS in tetrahydrofuran Standard solutions: Prepare a minimum of four Standard solutions in Diluent that bracket the expected clonidine related compound B concentration in the sample. The standard concentrations should be within the range of 0.210.0 g/mL. [NOTEStandard solutions are stable for up to 2 days if stored at 4.] Sample solution: Use the Sample solution, prepared as directed in the Assay. Analysis Samples: At least three Standard solutions that will bracket the expected sample concentration range and the Sample solution. Measure the responses for the clonidine related compound B. Calculate the peak response ratios of the analyte, and plot the results. Determine the linear regression equation of the standards by the mean-square method, and record the linear regression equation and the correlation coefficient; it should be NLT 0.995. Determine the concentration of the clonidine related compound B. Calculate the amount, in g/cm2, of the clonidine related compound B in the portion of the Transdermal System taken: Result = CV/A = concentration of clonidine related compound B from the linear regression analysis (g/mL) V = volume of the Sample solution (mL) A = area of the sample system (cm2) Acceptance criteria Clonidine related compound B: NMT 10.0 g/cm2 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in sealed, single-dose containers at a temperature not exceeding 30. LABELING: The label states the total amount of clonidine in the Transdermal System and the release rate, in mg/day, for the duration of the application of one system. USP REFERENCE STANDARDS 11 USP Clonidine RS USP Clonidine Related Compound B RS C
= concentration of clonidine in the sample of the standard curve (g/mL) = volume of the Medium (mL)
suitable cellulose membrane is available as Cuprophan 80M from Membrana GmbH, Oehder Strasse 28, D-42289, Wuppertal, Germany, fax number +49 02 02 60 57 15.
USP 32
Official Monographs / Clopidogrel 143

Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H16ClNO2S H2SO4 in the portion taken: Result = (rU/rS) (CS/CU) 100 rU = peak response from the Sample solution = peak response from the Standard solution rS CS = concentration of the Standard solution (mg/mL) = concentration of Sample solution (mg/mL) CU Acceptance criteria: 97.0%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE [NOTEFor all clopidogrel related compounds, the concentrations are expressed as bisulfate salts. Use bisulfate salt equivalents stated on USP Reference Standards labels to calculate the concentrations as appropriate.] Solution A, Mobile phase, and System suitability solution: Proceed as directed in the Assay. Standard stock solution: 20 g/mL of USP Clopidogrel Bisulfate RS, 40 g/mL of USP Clopidogrel Related Compound A RS, 120 g/mL of USP Clopidogrel Related Compound B RS, and 200 g/mL of USP Clopidogrel Related Compound C RS in methanol Standard solution: 0.5 g/mL of Clopidogrel Bisulfate, 1 g/mL of Clopidrogrel Related Compound A, 3 g/mL of Clopidrogrel Related Compound B, and 5 g/mL of Clopidrogrel Related Compound C from the Standard stock solution in Mobile phase Sample solution: Transfer about 100 mg of Clopidogrel Bisulfate to a 200-mL volumetric flask, dissolve in 5 mL of methanol, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L57 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clopidogrel related compound A, the two enantiomers of clopidogrel related compound B, clopidogrel, and clopidogrel related compound C are 0.5, 0.8, 1.2, 1.0, and 2.0, respectively.] Suitability requirements Resolution: Greater than 2.5 between clopidogrel and the first enantiomer of clopidogrel related compound B Relative standard deviation: NMT 15% for each peak, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of clopidogrel related compound A and clopidogrel related compound C in the portion of C16H16CINO2S H2SO4 taken: Result = (rU/rS) (CS1/CU) 100 rU rS = relevant clopidogrel response from the = relevant clopidogrel response from the related compound peak Sample solution related compound peak Standard solution
Clopidogrel Bisulfate
(Comment on this Monograph)id=m18680=Clopidogrel Bisulfate=Chl-Cy-Monos.pdf)
C16H16ClNO2S H2SO4 419.90 Thieno[3,2-c]pyridine-5(4H)-acetic acid, -(2-chlorophenyl)-6,7dihydro-, methyl ester, (S)-, sulfate (1:1); Methyl (+)-(S)- -(o-chlorophenyl)-6,7-dihydrothieno[3,2c]pyridine-5(4H)-acetate, sulfate (1:1) [120202-66-6]. DEFINITION Clopidogrel Bisulfate contains NLT 97.0% and NMT 101.5% of C16H16ClNO2S H2SO4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Sulfate 191: It responds to the test. ASSAY PROCEDURE [NOTEFor all clopidogrel related compounds, the concentrations are expressed as bisulfate salts. Use bisulfate salt equivalents stated on USP Reference Standards labels to calculate the concentrations as appropriate.] Solution A: 1.36 mg/mL of monobasic potassium phosphate in water Mobile phase: Acetonitrile and Solution A (1:3) System suitability solution: 100 g/mL of USP Clopidogrel Bisulfate RS and 200 g/mL of USP Clopidogrel Related Compound B RS in methanol Transfer 5 mL of this solution to a 200-mL volumetric flask, and dilute with Mobile phase to volume. Standard stock solution: 1.0 mg/mL of USP Clopidogrel Bisulfate RS in methanol Standard solution: Prepare 0.1 mg/mL from the Standard stock solution in Mobile phase Sample stock solution: 1 mg/mL of Clopidogrel Bisulfate in methanol Sample solution: 0.1 g/mL from Sample stock solution in Mobile phase Chromatographic system (See Chromatography 621 System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L57 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for the two enantiomers of clopidogrel related compound B and for clopidogrel are 0.8, 1.2, and 1.0, respectively.] Suitability requirements Resolution: Greater than 2.5 between clopidogrel and the first enantiomer of clopidogrel related compound B Relative standard deviation: NMT 1.0% from clopidogrel bisulfate, Standard solution
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= concentration of the relevant clopidogrel related compound in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Calculate the percentage of the first enantiomer of clopidogrel related compound B in the portion of C16H16CINO2 H2SO4 taken: Result = (rU/rS) (CS2/CU) 0.5 100 = first enantiomer of clopidogrel related compound B peak response from the Sample solution = first enantiomer of clopidogrel related rS compound B peak response from the Standard solution = concentration of clopidogrel related compound CS2 B in the Standard solution (mg/mL) = concentration of Sample solution (mg/mL) CU 0.5 = correction factor for the content of the first enantiomer in the clopidogrel related compound B Calculate the percentage of any impurity other than clopidogrel related compounds A, B, and C in the portion of C16H16CINO2 H2SO4 taken: Result = (rU/rS) (CS3/CU) 100 rU CS1
USP 32
Clopidogrel Tablets
(Comment on this Monograph)id=m18685=Clopidogrel Tablets=Chl-Cy-Monos.pdf) DEFINITION Clopidogrel Tablets contain Clopidogrel Bisulfate equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of clopidogrel (C16H16ClNO2S). IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 250300 nm Solution: Use the Sample solution prepared as directed in the test for Uniformity of Dosage Units. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE [NOTEFor all clopidogrel related compounds, the concentrations are expressed as bisulfate salts. Use bisulfate salt equivalents stated on USP Reference Standards labels to calculate the concentrations as appropriate.] Solution A: 1.36 mg/mL of monobasic potassium phosphate in water Mobile phase: Acetonitrile and Solution A (1:3) System suitability solution: 100 g/mL of USP Clopidogrel Bisulfate RS and 200 g/mL of USP Clopidogrel Related Compound B RS in methanol Transfer 5 mL of this solution to a 200-mL volumetric flask, and dilute with Mobile phase to volume. Standard solution: 0.1 mg/mL of USP Clopidogrel Bisulfate RS in methanol Sample solution: Weigh and finely powder NLT 20 Tablets. Mix the equivalent to 75 mg of clopidogrel (base) from the powder with 50 mL of methanol. Sonicate for 5 min, and stir for 30 min. Dilute with methanol to 100 mL. Transfer 5.0 mL of this solution to a 50-mL volumetric flask, and dilute with methanol to volume. Filter through a 0.45-m or finer porosity filter, and use the filtrate after discarding the first 5 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L57 Flow rate: 1.0 mL/min Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for the two enantiomers of clopidogrel related compound B and clopidogrel are 0.8 and 1.2; and 1.0, respectively.] Suitability requirements Resolution: Greater than 2.5 between clopidogrel and the first enantiomer of clopidogrel related compound B Relative standard deviation: NMT 1.0% from clopidogrel bisulfate, Standard solution
= peak response of any other impurity from the Sample solution = clopidogrel peak response from the Standard rS solution = concentration of clopidogrel bisulfate in the CS3 Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU [NOTEDisregard any peak observed in the blank.] Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 1.5% rU
Impurity Table 1 Name Clopidogrel related compound A First enantiomer of clopidogrel related compound B Clopidogrel related compound C Any other impurity Acceptance Criteria, NMT (%) 0.2 0.3 1.0 0.1
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Clopidogrel Bisulfate RS USP Clopidogrel Related Compound A RS USP Clopidogrel Related Compound B RS USP Clopidogrel Related Compound C RS
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H16ClNO2S H2SO4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) (100/L) = peak response from the Sample solution rU = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (tablet/mL) CU = molecular weight of clopidogrel, 321.82 Mr1 = molecular weight of clopidogrel bisulfate, 419.90 Mr2 L = label claim (mg/tablet) Acceptance criteria: 90.0%110.0% IMPURITIES Organic Impurities PROCEDURE [NOTEFor all clopidogrel related compounds, the concentrations are expressed as bisulfate salts. Use bisulfate salt equivalents stated on USP Reference Standards labels to calculate the concentrations as appropriate.] Solution A and Mobile phase: Proceed as directed in the Assay. System suitability solution: 100 g/mL of USP Clopidogrel Bisulfate RS and 200 g/mL of USP Clopidogrel Related Compound B RS in methanol Transfer 5 mL of this solution to a 200-mL volumetric flask, and dilute with Mobile phase to volume. Standard stock solution: 40 g/mL of USP Clopidogrel Bisulfate RS, 250 g/mL of USP Clopidogrel Related Compound A RS and 300 g/mL of USP Clopidogrel Related Compound C RS in methanol Standard solution 1 g/mL of clopidogrel bisulfate, 6 g/mL clopidogrel related compound A, and 7.5 g/mL clopidogrel related compound C from the Standard stock solution in Mobile phase Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer the equivalent to 75 mg of clopidogrel (free base) from powder to a 200-mL volumetric flask, add 5 mL of methanol, and dilute with Mobile phase to volume. Allow to stand for 10 min, and mix. Filter through a 0.45m or finer porosity filter, and use the filtrate after discarding the first 5 mL. Chromatographic system Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L57 Flow rate: 1.0 mL/min Injection size: 10 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clopidogrel related compound A, the two enantiomers of clopidogrel related compound B, clopidogrel related compound C, and clopidogrel are 0.5; 0.8 and 1.2, 1.0; and 2.0, respectively.] Suitability requirements Resolution: Greater than 2.5 between clopidogrel and the first enantiomer of clopidogrel related compound B Relative standard deviation: NMT 15% for each peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of clopidogrel related compound A and clopidogrel related compound C in the portion of Tablets taken: Result = (rU/rS) (CS/W) (Mr1/Mr2) 20 rU = peak response for the relevant clopidogrel related compound from the Sample solution rS
Official Monographs / Clopidogrel 145

= peak response for the relevant clopidogrel related compound from the Standard solution CS = concentration of the relevant clopidogrel related compound in the Standard solution (g/mL) W = weight of clopidogrel in the portion of Tablets used to prepare the Sample solution based on the labled quantity of clopidogrel per Tablet, Tablet weight and the weight of the portion of Tablets used (g) Mr1 = molecular weight of clopidogrel, 321.82 Mr2 = molecular weight of clopidogrel bisulfate, 419.90 Calculate the percentage of any impurity other than clopidogrel related compounds B in the portion of Tablets taken: Result = (rU/rS) (C/W) (Mr1/Mr2) 20 = peak response of any other impurity from the Sample solution rS = peak response of the clopidogrel peak from the Standard solution C = concentration of clopidogrel bisulfate in the Standard solution (g/mL) W = weight of clopidogrel in the portion of Tablets used to prepare the Sample solution based on the labled quantity of clopidogrel per Tablet, Tablet weight and the weight of the portion of Tablets used (g) = molecular weight of clopidogrel, 321.82 Mr1 = molecular weight of clopidogrel bisulfate, 419.90 Mr2 Acceptance criteria Individual impurities: See Impurity Table1. Total impurities (excluding clopidogrel related compound B): NMT 2.5%
Impurity Table 1 Acceptance Criteria, NMT (%) 1.2 1.5 0.2
rU
Impurity name Clopidogrel related compound A Clopidogrel related compound C Any other single impurity (excluding clopidogrel related compound B)
PERFORMANCE TESTS DISSOLUTION 711 Medium: pH 2.0 hydrochloric acid buffer (see Reagents, Indicators, and Solutions, Buffer solutions); 1000 mL Apparatus 2: 50 rpm Time: 30 min Detector: UV 240 nm Sample solutions: Sample per Dissolution 711 Standard solution: Dissolve USP Clopidogrel Bisulfate RS in 20.0 mL of methanol, and dilute quantitatively, and stepwise if necessary, with Medium to obtain a solution having a known concentration corresponding to that of the solution under test. Tolerances: NLT 80% (Q) of the labeled amount of C16H16 ClNO2S is dissolved in 30 min. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Content Uniformity Sample solution: Place 1 Tablet in 50.0 mL of 0.1 N hydrochloric acid. Sonicate for 5 min, and cool. Quantitatively transfer 5.0 mL of this solution to a 50-mL volumetric flask, and dilute with 0.1 N hydrochloric acid. Pass a portion of this solution through a suitable filter having a 0.45-m or finer porosity, and discard the first 5 mL of the filtrate.
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Adjust with 2.5 N sodium hydroxide to a pH of 8.0. Mobile phase: Acetonitrile, water, and Solution A (4:5:1) (See Chromatography 621, System Suitability.) Internal standard solution: Dissolve 5 mL of 2,6dimethylaniline in 50 mL of hexane, and carefully add dropwise hydrochloric acid to precipitate the amine hydrochloride. Filter through a sintered-glass funnel, wash the solid precipitate with hexane, and allow the precipitate to dry. Transfer about 50 mg of the dried precipitate of 2,6-dimethylaniline hydrochloride to a 100-mL volumetric flask, add 10.0 mL of Solution A and 40 mL of water, and dilute with acetonitrile to volume. Standard solution: 75 g/mL of USP Nordazepam RS in acetonitrile Transfer 4.0 mL of this solution to a 50-mL conical flask, then add 4.0 mL of 0.7 M potassium carbonate, 2.0 mL of Internal standard solution, and 15.0 mL of water. Insert a stopper, and mix. Sample solution: Transfer about 50 mg of Clorazepate Dipotassium to a 50-mL conical flask. Add 4.0 mL of 0.7 M potassium carbonate, and start stirring the solution. Add 2 mL of Internal standard solution and 19.0 mL of water. Stop stirring about 5 min after the addition of the 0.7 M potassium carbonate solution. [NOTEPrepare fresh immediately before each injection.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 232 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.0 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for 2,6-dimethylaniline and nordazepam are 0.8 and 1.0, respectively.] Suitability requirements Relative standard deviation: NMT 2.0% for the peak area ratio of nordazepam to 2,6-dimethylaniline Analysis Samples: Standard solution and Sample solution. Calculate the percentage of nordazepam in the portion of Clorazepate Dipotassium taken: Result = (RU/RS) (CS/CU) 100 = peak area ratio of any impurity to 2,6dimethylaniline of the Sample solution RS = peak area ratio of nordazepam to 2,6dimethylaniline of the Standard solution CS = concentration of USP Nordazepam RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: Nordazepam: NMT 0.5% Any individual impurity: NMT 0.1% Procedure 2 Diluent: Acetonitrile and 0.001 N sodium hydroxide (1:1) Mobile phase: Acetonitrile, 1 M solution of tetrabutylammonium hydroxide in methanol, and water (90:1:110) Adjust with phosphoric acid to a pH of 7.7. (See Chromatography 621, System Suitability.) Standard solution: 2.6 g/mL of USP 2-Amino-5chlorobenzophenone RS in Diluent RU
Standard solution: USP Clopidogrel Bisulfate RS in 0.1 N hydrochloric acid Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 270 nm Cell: 1 cm Analysis: Determine the amount of clopidogrel by employing UV absorption of the Standard solution and the Sample solution at the wavelength of maximum absorbance. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Clopidogrel Bisulfate RS USP Clopidogrel Related Compound A RS USP Clopidogrel Related Compound B RS USP Clopidogrel Related Compound C RS
Clorazepate Dipotassium
(Comment on this Monograph)id=m18720=Clorazepate Dipotassium=Chl-Cy-Monos.pdf)
408.92 C16H11ClK2N2O4 1H-1,4-Benzodiazepine-3-carboxylic acid, 7-chloro-2,3dihydro-2-oxo-5-phenyl-, potassium salt compound with potassium hydroxide (1:1); Potassium 7-chloro-2,3-dihydro-2-oxo-5-phenyl-1H-1,4benzodiazepine-3-carboxylate compound with potassium hydroxide (1:1) [57109-90-7]. DEFINITION Clorazepate Dipotassium contains NLT 98.5% and NMT 101.5% of C16H11ClK2N2O4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Medium: 0.4 mg/mL of sodium hydroxide solution Solution: 7 g/mL ASSAY PROCEDURE Sample solution: 150 mg of Clorazepate Dipotassium in 100 mL of glacial acetic acid Analysis: Titrate with 0.1 N perchloric acid VS, determining the endpoint potentiometrically, using a glass electrode and a calomel electrode containing a 1-100 solution of lithium perchlorate in glacial acetic acid. Perform a blank determination (see Titrimetry 541), and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 13.63 mg of C16H11ClK2N2O4. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities Procedure 1 Solution A: 27.6 g/L of monobasic sodium phosphate in water
USP 32
Sample solution: Transfer 300 mg of Clorazepate Dipotassium to a glass test tube. Add 10.0 mL of Diluent, and vigorously mix on a vortex mixer for about 90 s. [NOTEPrepare fresh immediately before each injection.] Chromatographic system Mode: LC Detector: UV 238 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2.0 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 3.0% from the peak height Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Clorazepate Dipotassium taken: Result = (rU/rS) (CS/CU) 100 = peak height of each impurity of the Sample solution = peak height of 2-amino-5-chlorobenzophenone rS of the Standard solution = concentration of USP 2-Amino-5CS chlorobenzophenone RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 2-amino-5-chlorobenzophenone: NMT 0.1% Any other individual impurity: NMT 0.1% Total impurities from Procedure 1 and Procedure 2: NMT 1.0% rU SPECIFIC TESTS LOSS ON DRYING 731: Dry it in vacuum at 60 for 1 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve under nitrogen in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Clorazepate Dipotassium RS USP Nordazepam RS
Official Monographs / Clorazepate 147

Mobile phase: Acetonitrile and Solution A (3:7) Standard solution: 60 g/mL of USP Clorazepate Dipotassium RS in 0.01 M sodium hydroxide [NOTEShake by mechanical means for 15 min, and filter.] Sample stock solution: Weigh and finely powder NLT 20 Tablets. Transfer to a suitable container a portion equivalent to 75 mg of clorazepate dipotassium from powder, add 200 mL of 0.01 M sodium hydroxide, and homogenize for NLT 3 min. Sample solution Transfer 15 mL of Sample stock solution to a 100-mL volumetric flask, dilute with 0.01 M sodium hydroxide to volume, mix, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 230 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H11ClK2N2O4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 peak response of the Sample solution peak response of the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS CU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.01 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Analytical wavelength: UV 240 nm Standard solution: USP Clorazepate Dipotassium RS in Medium Sample solution: Sample per Dissolution 711 Dilute with Medium to a concentration similar to that of the Standard solution. Tolerances: NLT 80% (Q) of the labeled amount of C16H11ClK2N2O4 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Standard solution: 7.6 g/mL of USP Clorazepate Dipotassium RS in 0.01 M sodium hydroxide Sample solution: Transfer 1 Tablet to a suitable container, add 200 mL of 0.01 M sodium hydroxide, and homogenize for NLT 3 min. Centrifuge a portion of this solution for 15 min, and filter the supernatant, discarding the first 20 mL. Prepare a 7.6 g/mL solution by diluting the filtrate with 0.01 M sodium hydroxide. Blank: 0.01 M sodium hydroxide Mode: Spectrophotometry Analytical wavelength: 231 nm Cell: 1 cm Analysis: Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. = = = =
Clorazepate Dipotassium Tablets

(Comment on this Monograph)id=m18760=Clorazepate Dipotassium Tablets=Chl-Cy-Monos.pdf) DEFINITION Clorazepate Dipotassium Tablets contain NLT 90.0% and NMT 110.0% of C16H11ClK2N2O4. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Transfer 5.0 mL of 1 M tetrabutylammonium hydroxide in methanol to a 1-L volumetric flask, dilute with water to volume, and adjust with phosphoric acid to a pH of 7.5.
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Mode: LC Detector: UV 238 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2.0 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution. Calculate the percentage of each impurity in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of each impurity from the Sample solution = peak response of the 2-amino-5rS chlorobenzophenone peak from the Standard solution = concentration of USP 2-Amino-5CS chlorobenzophenone RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: NMT 0.5% for the sum of all impurities, other than nordazepam from Procedure 1 and Procedure 2 rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP 2-Amino-5-chlorobenzophenone RS USP Clorazepate Dipotassium RS USP Nordazepam RS
IMPURITIES Organic Impurities PROCEDURE 1 Solution A: 27.6 g/L of monobasic sodium phosphate in water Adjust with 1 N sodium hydroxide to a pH of 8.0. Mobile phase: Acetonitrile, water, and Solution A (4:5:1) Standard solution: Transfer 4.0 mL of 66 g/mL of USP Nordazepam RS in acetonitrile to a 25-mL volumetric flask, add 5.0 mL of 0.7 M potassium carbonate and 3.0 mL of acetonitrile, dilute with water to volume, mix, and filter. Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion equivalent to 15 mg of clorazepate dipotassium from powder to a suitable container. Add 5 mL of acetonitrile, 5 mL of 0.7 M potassium carbonate, and 15 mL of water, stir for 10 min, and filter. [NOTEPrepare fresh before each injection, and use within 3 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 232 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution. Record the chromatograms for NLT twice the retention time of nordazepam, and measure the peak responses. Calculate the percentage of each impurity in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response of each impurity from the Sample solution rS = peak response for nordazepam from the Standard solution = concentration of USP Nordazepam RS in the CS Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: NMT 2.0% for nordazepam PROCEDURE 2 Diluent: Acetonitrile and 0.1 mM sodium hydroxide (3:7) Mobile phase: Acetonitrile, 1 M solution of tetrabutylammonium hydroxide in methanol, and water (90:1:110) Adjust with phosphoric acid to a pH of 7.7. Standard stock solution A: 0.5 mg/mL USP 2-Amino-5chlorobenzophenone RS in acetonitrile Standard stock solution B: 0.25 mg/mL USP 2-Amino-5chlorobenzophenone RS from Standard stock solution A in water Standard solution: 7.5. g/mL USP 2-Amino-5chlorobenzophenone RS from Standard stock solution B in Diluent Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion equivalent to 15 mg of clorazepate dipotassium from powder to a suitable container, add 10 mL of Diluent, shake by mechanical means for 10 min, and filter. Chromatographic system (See Chromatography 621, System Suitability.) rU
Clorsulon
(Comment on this Monograph)id=m18790=Clorsulon=Chl-CyMonos.pdf)
380.66 C8H8Cl3N3O4S2 1,3-Benzenedisulfonamide, 4-amino-6-(trichloroethenyl)-; 4-Amino-6-(trichlorovinyl)-m-benzenedisulfonamide [60200-06-8]. DEFINITION Clorsulon contains NLT 98.0% and NMT 101.0% of C8H8Cl3N3O4S2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M: Meets the requirements B. The retention time of the major peak for clorsulon from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
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ASSAY PROCEDURE [NOTEThe Standard and Sample solutions should be stored in low-actinic glassware.] Mobile phase: Acetonitrile, glacial acetic acid, and water (30:0.1:70) Standard solution: 0.1 mg/mL of USP Clorsulon RS in Mobile phase Sample stock solution: 1 mg/mL of Clorsulon in Mobile phase Sample solution: 0.1 g/mL from the Sample stock solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 30 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 7400 theoretical plates Tailing factor: NMT 1.4 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C8H8Cl3N3O4S2 in the portion of Clorsulon taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution rU = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 30 ppm Organic Impurities PROCEDURE [NOTEThe Standard and Sample solutions should be stored in low-actinic glassware.] Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution A: 10 mg/mL of USP Clorsulon RS in methanol Standard solution B: 1 mg/mL from Standard solution A in methanol Sample solution: 10 mg/mL of Clorsulon in methanol Developing solvent system: Chloroform and methanol (4:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Analysis: Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Apply 10-L portions of the Sample solution and of Standard solution A, and 5- and 10L portions of Standard solution B. Allow the spots to dry, and develop the chromatograms in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, allow the solvent to evaporate, and examine the plate under shortwavelength UV light: the chromatograms show principal spots at the same RF value. Estimate the amounts of any additional spots observed from the Sample solution by comparing them with the spots in the two
Official Monographs / Clotrimazole 149

chromatograms from Standard solution B, corresponding to 0.5% and 1.0% of impurity. Acceptance criteria Any individual impurities: NMT 0.5% Total impurities: NMT 2.0% SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 197203 LOSS ON DRYING 731: Dry it in vacuum at 100 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Clorsulon RS
Clotrimazole
(Comment on this Monograph)id=m18880=Clotrimazole=ChlCy-Monos.pdf)
C22H17ClN2 344.84 1H-Imidazole, 1-[(2-chlorophenyl)diphenylmethyl]-; 1-(o-Chloro-,-diphenylbenzyl)imidazole [23593-75-1]. DEFINITION Clotrimazole contains NLT 98.0% and NMT 102.0% of C22H17ClN2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M: Meets the requirements B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 20 mg/mL in chloroform Developing solvent system: Xylene, n-propyl alcohol, and ammonium hydroxide (180:20:1) ASSAY PROCEDURE Solution A: 4.35 mg/mL of dibasic potassium phosphate Mobile phase: Methanol and Solution A (3:1) Pass through a membrane filter having a 0.2 m or finer porosity. The ratio of volumes may be changed to obtain the required resolution. Internal standard solution: Transfer 33 mg of testosterone propionate to a 200-mL volumetric flask, dissolve in 125 mL of methanol, add 50 mL of Solution A, and dilute with methanol to volume (165 g/mL of testosterone propionate). Standard stock solution A: Transfer 50 mg of USP Clotrimazole RS to a 50-mL volumetric flask. Add 25 mL of methanol to dissolve, add 12.5 mL of Solution A, and dilute with methanol to volume (1 mg/mL of USP Clotrimazole RS). Standard solution: Transfer 10.0 mL of Standard stock solution to a 100-mL volumetric flask, add 4.0 mL of Internal standard solution, and dilute with Mobile phase to volume. Standard stock solution B: Transfer 12.5 mg of USP Clotrimazole Related Compound ARS to a 25 mL volumetric flask, add 10 mL of methanol to dissolve, add 6.25 mL of Solution A, and dilute with methanol to volume.
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PROCEDURE 2: LIMIT OF CLOTRIMAZOLE RELATED COMPOUND A Solution A, Mobile phase, Standard stock solution B, System suitability solution, and Chromatographic system: Proceed as directed in the Assay. Standard solution: Transfer 5.0 mL of the Standard stock solution B to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution: Sample stock solution from the Assay Analysis Samples: Standard solution and Sample solution Calculate the percentage of clotrimazole related compound A in the portion of Clotrimazole taken: Result = (rU/rS) (CS/CU) 100 = peak response for clotrimazole related compound A from the Sample solution = peak response for clotrimazole related rS compound A from the Standard solution = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: NMT 0.5% rU SPECIFIC TESTS LOSS ON DRYING 731: 0.5% of its weight. Dry it at 105 for 2 h: it loses NMT
System suitability solution: Transfer 3 mL of Standard stock solution A and 5 mL of Standard stock solution B to a 25-mL volumetric flask, and dilute with Mobile phase to volume. Sample stock solution: Transfer 100 mg of Clotrimazole to a 10 mL volumetric flask, add 5 mL of methanol to dissolve, add 2.5 mL of Solution A, and dilute with methanol to volume. Sample solution: Transfer 1.0 mL of the Sample stock solution to a 100-mL volumetric flask, add 4.0 mL of Internal standard solution, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 10 m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution Suitability requirements [NOTEThe relative retention times for clotrimazole related compound A, clotrimazole, and testosterone propionate are 0.7, 1.0, and 1.5, respectively.] Resolution: NLT 1.9 between clotrimazole and clotrimazole related compound A, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 in the portion of Clotrimazole taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clotrimazole to testosterone propionate from the Sample solution = peak response ratio of clotrimazole to RS testosterone propionate from the Standard solution CS = concentration of the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 98.0%02.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE 1: LIMIT OF IMIDAZOLE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 500 g/mL of USP Imidazole RS in chloroform Sample solution: 100 mg/mL of Clotrimazole in chloroform Application volume: 5 L Developing solvent system: Methanol and chloroform (3:2) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. After air-drying the plate for 5 min, place it in a closed container with a dish containing 100 g of iodine in a shallow layer, and allow to remain for 60 min. Remove the plate from the container, and observe the chromatogram. Acceptance criteria: Any brown spot obtained from the Sample solution at an RF value corresponding to the principal spot from the Standard solution is not greater in size or intensity than the principal spot from the Standard solution: NMT 0.5% of imidazole. RU
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Clotrimazole RS USP Clotrimazole Related Compound A RS USP Imidazole RS
Clotrimazole Cream
(Comment on this Monograph)id=m18890=Clotrimazole Cream=Chl-Cy-Monos.pdf) DEFINITION Clotrimazole Cream contains NLT 90.0% and NMT 110.0% of the labeled amount of clotrimazole (C22H17ClN2). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 1 mg/mL of USP Clotrimazole RS in chloroform Sample solution: Transfer the equivalent to 5 mg of clotrimazole from Cream to a 50-mL centrifuge tube, add 5 mL of chloroform, and centrifuge to obtain a clear solution. Solution A: 30 mg/mL of bismuth subnitrate and 300 mg/mL of potassium iodide in dilute hydrochloric acid (1 in 4) Dilute 10 mL of this solution with water to 100 mL. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Spray reagent: Dilulte 10 mL of Solution A, and 5 mL of dilute hydrochloric acid (1 in 4) with water to 200 mL. Analysis Samples: Standard solution and Sample solution In a suitable chromatographic chamber containing 200 mL of ether, place a beaker containing 25 mL of ammonium hydroxide. Cover the chamber, and allow to equilibrate for 2 h. Develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate
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the spots on the plate by examination under shortwavelength UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. Spray the plate evenly with Spray reagent: the principal spots from the Sample solution and Standard solution are orange. ASSAY PROCEDURE Solution A: 4.35 mg/mL of dibasic potassium phosphate Mobile phase: Methanol and Solution A (3:1) [NOTEThe ratio of volumes may be changed to obtain the required resolution.] Internal standard solution: 0.07 mg/mL of testosterone propionate in dehydrated alcohol Standard stock solution A: 1 mg/mL of USP Clotrimazole RS in dehydrated alcohol Standard stock solution B: 0.12 mg/mL of USP Clotrimazole Related Compound A RS in dehydrated alcohol Standard solution: Standard stock solution A and Internal standard solution (1:1) System suitability solution: Standard stock solution A and Standard stock solution B (1:7) Sample solution: Transfer the equivalent to 10 mg of clotrimazole from Cream to a 50-mL screw-capped centrifuge tube. Add 10.0 mL of Internal standard solution, and heat at 50 in a water bath for 5 min, with occasional shaking. Remove the tube from the bath, and shake vigorously for 5 min. Cool in a methanol-ice bath for 15 min, and promptly centrifuge. Transfer the supernatant to a test tube. Add 10.0 mL of dehydrated alcohol to the residue in the centrifuge tube, and repeat the extraction starting with heat at 50 in a water bath. Transfer the supernatant to the test tube containing the supernatant from the first extraction. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Guard column: 2.1-mm 6-cm; 10 m packing L7 Column: 3.9-mm 30-cm; 10 m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for clotrimazole related compound A, clotrimazole, and testosterone propionate are 0.9, 1.0, and 1.5, respectively.] Suitability requirements Resolution: NLT 1.2 between clotrimazole related compound A and clotrimazole, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 in each g of Cream taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = peak response ratio of clotrimazole to testosterone propionate from the Sample solution = peak response ratio of clotrimazole to testosterone propionate from the Standard solution = concentration of USP Clotrimazole RS in the Standard solution (mg/mL) = nominal concentration of clotrimazole in the Sample solution (mg/mL)

ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers, at a temperature between 2 and 30. LABELING: Cream that is packaged and labeled for use as a vaginal preparation shall be labeled Clotrimazole Vaginal Cream. USP REFERENCE STANDARDS 11 USP Clotrimazole RS USP Clotrimazole Related Compound A RS
Clotrimazole Lotion
(Comment on this Monograph)id=m18897=Clotrimazole Lotion=Chl-Cy-Monos.pdf) DEFINITION Clotrimazole Lotion contains NLT 90.0% and NMT 110.0% of the labeled amount of clotrimazole (C22H17ClN2). IDENTIFICATION The retention time of the major peak for clotrimazole from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 4.35 mg/mL of dibasic potassium phosphate in water Mobile phase: Methanol and Solution A (3:1)[NOTEThe ratio of volumes may be changed to obtain the required resolution.] Internal standard solution: 0.07 mg/mL of testosterone propionate in dehydrated alcohol Standard stock solution A: 2 mg/mL of USP Clotrimazole RS in dehydrated alcohol Standard stock solution B: 0.1 mg/mL of USP Clotrimazole Related Compound A RS in dehydrated alcohol Standard solution: Standard stock solution A, Standard stock solution B, and Internal standard solution (1:1:2) Sample solution: Transfer the equivalent of 10 mg of clotrimazole from freshly mixed Lotion to a screw-capped, 50-mL centrifuge tube. Add 10.0 mL of Internal standard solution, place the cap on the tube, and heat at 50 in a water bath for 5 min, with occasional shaking. Remove the tube from the bath, and shake vigorously for 5 min. Cool in a methanol-ice bath for 15 min, and promptly centrifuge. Transfer the supernatant to a test tube. Add 10.0 mL of dehydrated alcohol to the residue in the centrifuge tube, and repeat the extraction as directed above, beginning with place the cap on the tube. Transfer the supernatant to the test tube containing the supernatant from the first extraction. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 10 m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for clotrimazole related compound A, clotrimazole, and testosterone propionate are 0.9, 1.0, and 1.5, respectively.] Suitability requirements Resolution: NLT 1.2 between clotrimazole related compound A and clotrimazole, and NLT 1.9 between
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IDENTIFICATION A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 2.5 mg/mL of USP Clotrimazole RS in dichloromethane Sample solution: Place an equivalent to 50 mg of clotrimazile, from a quantity of finely powdered Lozenges, into a screw-capped 50-mL test tube, and add 20.0 mL of dichloromethane. Shake by mechanical means for 10 min, and allow the suspension to settle. Use the supernatant. Solution A: Dissolve 3 g of bismuth subnitrate and 30 g of potassium iodide in 10 mL of dilute hydrochloric acid (1 in 4), and dilute with water to 100 mL. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Developing solvent system: Diethyl ether and ammonium hydroxide (8:1) Spray reagent: Dilute 10 mL of Solution A and 10 mL of dilute hydrochloric acid (1 in 4) with water to 100 mL. Analysis Samples: Standard solution and Sample solution Proceed as directed in the chapter. Position the plate in a chromatographic chamber, and develop the chromatograms in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Examine the plate under short-wavelength UV light and then spray with the Spray reagent. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution, and after the plates have been sprayed with the Spray reagent, the clotrimazole spots are orange. ASSAY PROCEDURE Solution A: 1 mg/mL of ammonium carbonate in water. Adjust with 10% sulfuric acid solution to a pH of 6.0. Mobile phase: Methanol and Solution A (3:1) Internal standard stock solution: 5 mg/mL of triphenylmethane in methanol Internal standard solution: Pipet 10.0 mL of Internal standard stock solution into a 250-mL volumetric flask, and dilute with Mobile phase to volume. Standard solution: Transfer about 20 mg of USP Clotrimazole RS to a 100-mL volumetric flask. Add 4.0 mL of Internal standard stock solution, dissolve in and dilute with Mobile phase to volume. Sample solution: Transfer an equivalent to 5 mg of clotrimazole, from weighed and pulverized Lozenges (NLT 10), to a 50-mL screw-capped centrifuge tube. Pipet 25 mL of Internal standard solution into the tube. Sonicate for 10 min, then shake for 10 min. Centrifuge at 2500 rpm for 30 min. Use the clear supernatant layer. Chromatographic system (See Chromatography 621, System Suitability.)
clotrimazole and testosterone propionate in the Standard solution Relative standard deviation: NMT 2.0% in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 in each g of Lotion taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clotrimazole to testosterone propionate from the Sample solution = peak response ratio of clotrimazole to RS testosterone propionate from the Standard solution = concentration of USP Clotrimazole RS in the CS Standard solution (mg/mL) = nominal concentration of clotrimazole in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU IMPURITIES Organic Impurities LIMIT OF CLOTRIMAZOLE RELATED COMPOUND A Analysis: Using the chromatograms of the Standard solution and Sample solution as obtained in the Assay, calculate the percentage of clotrimazole related compound A in the portion of Lotion taken: Result = (RU/RS) (CS/CU) 100 RU = peak response ratio of clotrimazole related compound A to testosterone propionate from the Sample solution = peak response ratio of clotrimazole related RS compound A to testosterone propionate from the Standard solution = concentration of USP Clotrimazole Related CS Compound A RS in the Standard solution (mg/mL) CU = nominal concentration of clotrimazole related compound A in the Sample solution (mg/mL) Acceptance criteria: NMT 5.0%
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements for absence of Staphylococcus aureus and Pseudomonas aeruginosa. PH 791: 5.07.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a temperature between 2 and 30. USP REFERENCE STANDARDS 11 USP Clotrimazole RS USP Clotrimazole Related Compound A RS
Clotrimazole Lozenges
(Comment on this Monograph)id=m18899=Clotrimazole Lozenges=Chl-Cy-Monos.pdf) DEFINITION Clotrimazole Lozenges contain NLT 90.0% and NMT 110.0% of the labeled amount of clotrimazole (C22H17ClN2) in a suitable molded base.
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Mode: LC Detector: UV 215 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for clotrimazile and triphenylmethane are about 1.0 and 1.4, respectively.] Suitability requirements Resolution: NLT 1.5 between clotrimazole and triphenylmethane Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2 Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 in the portion of Lozenges taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clotrimazole to triphenylmethane peak from the Sample solution = peak response ratio of clotrimazole to RS triphenylmethane peak from the Standard solution CS = concentration of USP Clotrimazole RS in the Standard solution (mg/mL) = nominal concentration of clotrimazole in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 500 mL, deaerated Apparatus 2: 50 rpm Time: 45 min Determine the amount of C22H17ClN2 dissolved by employing the following method. Solution A: 4.4 mg/mL of dibasic potassium phosphate in water Solution B: 17.4 mg/mL of dibasic potassium phosphate in water Diluent: Methanol and Solution B (3:2) Mobile phase: Methanol and Solution A (4:1) Standard stock solution: 0.02 mg/mL of USP Clotrimazole RS in Medium Standard solution: 4 g/mL from the Standard stock solution in Diluent Sample solution: Withdraw 25 mL of the solution under test from the vessel. Pass through a 0.45-m polyvinylidene difluoride filter, discard the first 10 mL of the filtrate. Transfer 5.0 mL of filtrate to a 25 mL volumetric flask, and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) RU

Mode: LC Detector: UV 215 nm Column: 3.9-mm 7.5-cm; packing L1 Flow rate: 1.0 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 taken: Result = (rU/rS) (CS/L) D V 100 = peak response from the Sample solution rU = peak response from the Standard solution rS = concentration of the Standard solution (mg/mL) CS L = label claim of clotrimazole in 1 lozenge (mg) D = dilution factor for the Sample solution V = volume of Medium, 500 mL Tolerances: NLT 80% (Q) of the labeled amount of C22H17ClN2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clotrimazole RS
Clotrimazole Topical Solution

(Comment on this Monograph)id=m18920=Clotrimazole Topical Solution=Chl-Cy-Monos.pdf) DEFINITION Clotrimazole Topical Solution is a solution of Clotrimazole in a suitable nonaqueous, hydrophilic solvent. It contains NLT 90.0% and NMT 115.0% of the labeled amount of clotrimazole (C22H17ClN2). IDENTIFICATION PROCEDURE Standard solution: 1 mg/mL of USP Clotrimazole RS in chloroform Sample solution: Transfer a volume of Topical Solution, equivalent to 10 mg of clotrimazole, to a screw-capped, 50mL centrifuge tube. Add 5 mL of dilute ammonium hydroxide (1 in 100) and 10 mL of chloroform. Shake vigorously, and centrifuge to obtain a clear chloroform phase. Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Solution A: Dissolve 3 g of bismuth subnitrate and 30 g of potassium iodide in 10 mL of dilute hydrochloric acid (1 in 4), and dilute with water to 100 mL.
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RS
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= peak response ratio of clotrimazole to testosterone propionate from the Standard solution = concentration of the Standard solution (mg/mL) CS = nominal concentration of clotrimazole in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers at a temperature between 2 and 30. USP REFERENCE STANDARDS 11 USP Clotrimazole RS USP Clotrimazole Related Compound A RS
Spray reagent: Dilute 10 mL of Solution A and 10 mL of dilute hydrochloric acid (1 in 4) with water to 100 mL. Chromatographic chamber: 200 mL of ether Place a beaker containing 25 mL of ammonium hydroxide. Cover the chamber, and allow to equilibrate for 2 h. Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. Spray the plate evenly with the Spray reagent: the principal spots from the Sample solution and the Standard solution are orange. ASSAY PROCEDURE Solution A: 4.35 mg/mL of dibasic potassium phosphate in water Mobile phase: Methanol and Solution A (3:1) [NOTEThe ratio of volumes may be changed to obtain the required resolution.] Internal standard solution: Transfer 66 mg of testosterone propionate to a 100-mL volumetric flask, dissolve in 75 mL of methanol, and dilute with Solution A to volume. Standard solution: Transfer 50 mg of USP Clotrimazole RS to a 100-mL volumetric flask, dissolve in 75 mL of methanol, and dilute with Solution A to volume. System suitability stock solution: 0.2 mg/mL of USP Clotrimazole Related Compound A RS in methanol System suitability solution: Add 12.0 mL of System suitability stock solution to a 25-mL volumetric flask. Add 4 mL of Solution A and 3 mL of Standard solution, and dilute to volume with Mobile phase. Sample solution: Transfer a volume of Topical Solution, equivalent to 50 mg of clotrimazole, to a 50-mL volumetric flask. Add 5.0 mL of Internal standard solution, and dilute to volume with Mobile phase. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for clotrimazole related compound A, clotrimazole, and testosterone propionate are 0.9, 1.0, and 1.5, respectively.] Suitability requirements Resolution: NLT 1.2 between clotrimazole related compound A and clotrimazole, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 in each mL of Topical Solution taken: Result = (RU/RS) (CS/CU) 100 RU = peak response ratio of clotrimazole to testosterone propionate from the Sample solution
Clotrimazole Vaginal Inserts

(Comment on this Monograph)id=m18930=Clotrimazole Vaginal Inserts=Chl-Cy-Monos.pdf) DEFINITION Clotrimazole Vaginal Inserts contain NLT 90.0% and NMT 110.0% of the labeled amount of clotrimazole (C22H17ClN2). IDENTIFICATION PROCEDURE Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 5 mg/mL of USP Clotrimazole RS in chloroform Sample solution: Place an equivalent to 50 mg of clotrimazole, from finely powdered Vaginal Inserts, in a screw-capped 50-mL centrifuge tube. Add 10 mL of chloroform, and shake vigorously for 2 min. Centrifuge to clarify. [NOTEThe supernatant may remain slightly turbid.] Solution A: Dissolve 3 g of bismuth subnitrate and 30 g of potassium iodide in 10 mL of dilute hydrochloric acid (1 in 4), and dilute with water to 100 mL. Spray reagent: Dilute 10 mL of Solution A and 10 mL of dilute hydrochloric acid (1 in 4) with water to 100 mL. Application volume: 20 L Chromatographic chamber: 200 mL of ether Place a beaker containing 25 mL of ammonium hydroxide into the Chromatographic chamber. Cover the chamber, and allow to equilibrate for 2 h. Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by examining under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. Spray the plate evenly with this spray reagent: the principal spots from the Sample solution and the Standard solution are orange. ASSAY PROCEDURE Solution A: 4.35 mg/mL of dibasic potassium phosphate Mobile phase: Methanol and Solution A (3:1) [NOTEThe ratio of volumes may be changed to obtain the required resolution.]
USP 32
Internal standard solution: Transfer 66 mg of testosterone propionate to a 100-mL volumetric flask, dissolve in 75 mL of methanol, and dilute with Solution A to volume. Standard solution: Transfer 50 mg of USP Clotrimazole RS to a 100-mL volumetric flask, dissolve in 75 mL of methanol, and dilute with Solution A to volume. System suitability stock solution: 0.2 mg/mL of USP Clotrimazole Related Compound A RS in methanol System suitability solution: Add 12.0 mL of System suitability stock solution to a 25-mL volumetric flask, add 4 mL of Solution A and 3 mL of Standard solution, and dilute with Mobile phase to volume. Sample solution: Transfer an equivalent to 100 mg of clotrimazole from finely powdered Vaginal Inserts (NLT 10) to a 50-mL screw-capped centrifuge tube. Add 15 mL of Mobile phase and 10.0 mL of Internal standard solution, rotate for 15 min, and centrifuge for 10 min. Using a suitable syringe, transfer the supernatant to a 100-mL volumetric flask. Rinse the syringe with 25 mL of Mobile phase, adding the rinsings to the centrifuge tube. Rotate the centrifuge tube for 15 min, and centrifuge for 10 min. Using a suitable syringe, transfer the supernatant to the 100-mL volumetric flask. Rinse the syringe with 25 mL of Mobile phase, and add the washings to the 100-mL volumetric flask. Dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution and Standard solution [NOTEThe relative retention times for clotrimazole related compound A, clotrimazole, and testosterone propionate are about 0.9, 1.0, and 1.5, respectively.] Suitability requirements Resolution: NLT 1.2 between clotrimazole related compound A and clotrimazole, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 in the portion of Vaginal Inserts taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clotrimazole to testosterone propionate from the Sample solution = peak response ratio of clotrimazole to RS testosterone propionate from the Standard solution = concentration of USP Clotrimazole RS in the CS Standard solution (mg/mL) = nominal concentration of clotrimazole in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS DISINTEGRATION 701 Time: 20 min UNIFORMITY OF DOSAGE UNITS 905:
Clotrimazole and Betamethasone Dipropionate Cream

(Comment on this Monograph)id=m18935=Clotrimazole and Betamethasone Dipropionate Cream=Chl-Cy-Monos.pdf) DEFINITION Clotrimazole and Betamethasone Dipropionate Cream contains NLT 90.0% and NMT 110.0% of the labeled amount of clotrimazole (C22H17ClN2) and an amount of betamethasone dipropionate equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of betamethasone (C22H29FO5), in a suitable cream base. IDENTIFICATION The retention times of the major peaks for clotrimazole and betamethasone dipropionate from the Sample solution correspond to those of the Standard solution, as obtained in the Assay for clotrimazole, betamethasone, and limit of clotrimazole related compound A. ASSAY PROCEDURE Solution A: 6.6 mg/mL of dibasic ammonium phosphate in water Mobile phase: Prepare a mixture of methanol and Solution A (7:3), and adjust with phosphoric acid to a pH of 7.0 0.2. Pass through a membrane filter having a 0.45-m or finer porosity, and degas. Internal standard solution: 0.15 mg/mL of progesterone in alcohol Clotrimazole standard stock solution: 5 mg/mL of USP Clotrimazole RS in alcohol Betamethasone dipropionate standard stock solution: 6.4J mg/mL of USP Betamethasone Dipropionate RS in alcohol, J being the ratio of the labeled amount, in mg, of betamethasone to the labeled amount, in mg, of clotrimazole in each g of Cream Clotrimazole related compound A standard stock solution: 0.5 mg/mL of USP Clotrimazole Related Compound A RS in methanol Standard solution: Transfer 1.0 mL of Clotrimazole related compound A standard stock solution to a suitable container, and evaporate to dryness in a water bath at room temperature under a stream of nitrogen. To the residue, add 2.0 mL each of Clotrimazole standard stock solution, Betamethasone dipropionate standard stock solution, and Internal standard solution. Sample solution: Weigh a portion of Cream equivalent to 10 mg of clotrimazole, and transfer to a screwcapped, 50mL centrifuge tube. Add 2.0 mL of Internal standard solution and 4.0 mL of alcohol, place the cap on the tube, and heat at 60 in a water bath for 10 min, with occasional shaking. Remove the tube from the bath, cool in an ice bath for 20 min, and promptly centrifuge. Transfer a portion of the supernatant to a test tube, and use as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 10-m packing L1 Flow rate: 1.7 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention times for betamethasone dipropionate, clotrimazole related compound A, progesterone, and clotrimazole are about 1.0, 1.2, 1.4, and 1.7, respectively.]
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clotrimazole RS USP Clotrimazole Related Compound A RS
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers. USP REFERENCE STANDARDS 11 USP Betamethasone Dipropionate RS USP Clotrimazole RS USP Clotrimazole Related Compound A RS
Suitability requirements Resolution: NLT 1.0 between betamethasone dipropionate and clotrimazole related compound A, NLT 1.5 between clotrimazole related compound A and progesterone, and NLT 1.8 between progesterone and clotrimazole Relative standard deviation: NMT 2.0% determined from clotrimazole and betamethasone dipropionate, and NMT 4.0% determined from clotrimazole related compound A Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H17ClN2 taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clotrimazole to progesterone of the Sample solution = peak response ratio of clotrimazole to RS progesterone of the Standard solution = concentration of USP Clotrimazole RS in the CS Clotrimazole standard stock solution (mg/mL) CU = nominal concentration of clotrimazole in the Sample solution (mg/mL) Calculate the percentage of C22H29FO5 in each g of Cream taken: Result = (RU/RS) (CS/CU) (Mr1/Mr2) 100 RU = peak response ratio of betamethasone dipropionate to progesterone of the Sample solution RS = peak response ratio of betamethasone dipropionate to progesterone of the Standard solution CS = concentration of USP Betamethasone Dipropionate RS in the Betamethasone dipropionate standard stock solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) = molecular weight of betamethasone, 392.46 Mr1 = molecular weight of betamethasone Mr2 dipropionate, 504.60 Calculate the percentage of clotrimazole related compound A in each g of Cream taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of clotrimazole related compound A to progesterone of the Sample solution = peak response ratio of clotrimazole related RS compound A to progesterone of the Standard solution = concentration of USP Clotrimazole Related CS Compound A RS in the Clotrimazole related compound A standard stock solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% of the labeled amount of C22H17ClN2, 90.0%110.0% of the labeled amount of C22H29FO5, and NMT 5.0% for the limit of clotrimazole related compound A RU PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa RU
Cloxacillin Benzathine
(Comment on this Monograph)id=m18980=Cloxacillin Benzathine=Chl-Cy-Monos.pdf)
(C19H18ClN3O5S)2 C16H20N2 1112.11 4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[[[3-(2chlorophenyl)-5-methyl-4-isoxazolyl]carbonyl]amino]-3,3dimethyl-7-oxo-, [2S-(2,5,6)]-, compd. with N,Nbis(phenyl-methyl)-1,2-ethanediamine (2:1); (2S,5R,6R)-6-[3-(o-Chlorophenyl)-5-methyl-4isoxazolecarboxamido]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0]heptane-2-carboxylic acid compound with N,Ndibenzylethylenediamine (2:1) [23736-58-5]. DEFINITION Cloxacillin Benzathine has a potency equivalent to NLT 704 g and NMT 821 g of cloxacillin (C19H18ClN3O5S) per mg, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 20 mg in 5 mL of 5 N sodium hydroxide Heat on a steam bath for 20 min. Cool, mix 1 mL of this solution with 10 mL of 1.2 N sulfuric acid, and extract with 50 mL of ether. Wash the ether extract with 30 mL of water, and extract the ether layer with 50 mL of 0.1 N sodium hydroxide. Standard solution: Prepare a solution of 15 mg of USP Cloxacillin Sodium RS, following the same procedure as described for the Sample solution. ASSAY PROCEDURE Solution A: 13.8 g/L of monobasic sodium phosphate in water Mobile phase: Acetonitrile and Solution A (1:3) Adjust with phosphoric acid or 1 N sodium hydroxide to a pH of 4.6 0.2. Pass through a 0.45-m nylon filter, and degas. [NOTEThe retention time of cloxacillin is very sensitive to the acetonitrile content of the Mobile phase.] Diluent: 6.9 g/L of monobasic sodium phosphate in water Mix the resulting solution and acetonitrile (3:2). Adjust with phosphoric acid or 1 N sodium hydroxide to a pH of 6.4. Standard solutions: In duplicate, 112 g/mL of USP Cloxacillin Sodium RS in Diluent Sample solutions: In duplicate, 128 g/mL of Cloxacillin Benzathine in Diluent Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 10-m packing L1 Temperature: 40 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% [NOTEPeak areas of the two Standard solutions agree within 98%102%.] Analysis Samples: Standard solutions and Sample solutions Calculate the quantity, in g, of C19H18ClN3O5S in each mg of Cloxacillin Benzathine taken: Result = (rU/rS) (CS/CU) P = average peak area of cloxacillin from the Sample solutions = average peak area of cloxacillin from the rS Standard solutions = concentration of the Standard solutions (g/mL) CS = concentration of Cloxacillin Benzathine in the CU Sample solutions (g/mL) P = assigned potency of USP Cloxacillin Sodium RS (g of cloxacillin/mg) Acceptance criteria: 704821 g/mg SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 3.06.5, in 10 mg/mL suspension STERILITY TESTS 71: Where the label states that Cloxacillin Benzathine is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Direct Inoculation of the Culture Medium, except to use Fluid Thioglycollate Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, to use SoybeanCasein Digest Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, and to shake the tubes once daily. WATER DETERMINATION, Method I 921: NMT 5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that it is for veterinary use only. Where it is intended for use in preparing sterile dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of sterile dosage forms. USP REFERENCE STANDARDS 11 USP Cloxacillin Benzathine RS USP Cloxacillin Sodium RS rU
Official Monographs / Cloxacillin 157

IDENTIFICATION INFRARED ABSORPTION 197K Sample: Transfer a quantity of Intramammary Infusion, equivalent to 500 mg of cloxacillin, to a 50-mL centrifuge tube, add 25 mL of toluene, mix, and centrifuge. Decant and discard the toluene. Wash the residue with four 25-mL portions of toluene, sonicating for 30 s after each addition of toluene. Dry the residue in vacuum over silica gel. ASSAY PROCEDURE Solution A: Dissolve 55.2 g of monobasic sodium phosphate in water, and dilute with water to 4 L. Mobile phase: Acetonitrile and Solution A (1:3) Adjust with phosphoric acid or 1 N sodium hydroxide to a pH of 4.6 0.2. Pass through a 0.45-m nylon filter, and degas. [NOTEThe retention time of cloxacillin is very sensitive to the acetonitrile content of the Mobile phase.] Diluent: 6.9 mg/mL of monobasic sodium phosphate in water Mix the resulting solution and acetonitrile (3:2). Adjust with phosphoric acid or 1 N sodium hydroxide to a pH of 6.4. Standard solutions: In duplicate, 112 g/mL of USP Cloxacillin Sodium RS in Diluent Sample solutions: In duplicate, quantitatively express the entire contents of a syringe of Intramammary Infusion into a 500-mL volumetric flask. Add 300 mL of methanol, and stir for 45 min 1 min. Dilute with methanol to volume, and stir for an additional 10 min 1 min. Immediately transfer 45 mL of the resulting solution to a 50-mL polypropylene centrifuge tube, and centrifuge for 10 min. From the supernatant remove an aliquot, and dilute with a sufficient volume of Diluent to prepare a solution containing 100 g/mL of cloxacillin. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 10-m packing L1 Temperature: 40 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Peak areas of the two Standard solutions agree within 98% to 102%. Analysis Samples: Standard solutions and Sample solutions Calculate the percentage of C19H18ClN3O5S in each syringe of Intramammary Infusion taken: Result = (rU/rS) (CS/CU) P 100 rU = average peak areas of the cloxacillin peaks of the Sample solutions = average peak areas of the cloxacillin peaks of the Standard solutions = concentration of USP Cloxacillin Sodium RS in the Standard solutions (g/mL) = nominal concentration of cloxacillin in the Sample solutions (g/mL) = assigned potency of USP Cloxacillin Sodium RS (g of cloxacillin/mg)
Cloxacillin Benzathine Intramammary Infusion

(Comment on this Monograph)id=m18984=Cloxacillin Benzathine Intramammary Infusion=Chl-Cy-Monos.pdf) DEFINITION Cloxacillin Benzathine Intramammary Infusion is a suspension of Cloxacillin Benzathine in a suitable oil vehicle. It has a potency equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cloxacillin (C19H18ClN3O5S).
rS CS CU P
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90.0%120.0%
USP 32
Mobile phase: Acetonitrile and Solution A (2:8) Standard solution: 0.55 mg/mL of USP Cloxacillin Sodium RS in Solution A Sample solution: 110 mg of Cloxacillin Sodium to a 200mL volumetric flask, and dilute with Solution A to volume. Stir with the aid of a magnetic stirrer for 5 min to dissolve. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 225 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.8 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C19H18ClN3O5S in each mg of Cloxacillin Sodium taken: Result = (rU/rS) (CS/CU) E = peak response of cloxacillin from the Sample solution rS = peak response of cloxacillin from the Standard solution = concentration of USP Cloxacillin Sodium RS in CS the Standard solution (g/mL) CU = concentration of Cloxacillin Sodium in the Sample solution (g/mL) E = cloxacillin equivalent of USP Cloxacillin Sodium RS (g/mg) Acceptance criteria: NLT 825 g IMPURITIES DIMETHYLANILINE 223 Meets the requirement rU
SPECIFIC TESTS STERILITY TESTS 71 Where the label states that it is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Direct Inoculation of the Culture Medium, except to use Fluid Thioglycollate Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, to use SoybeanCasein Digest Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, and to shake the tubes once daily. WATER DETERMINATION, Method I 921: NMT 1.0%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in disposable syringes that are well-closed containers, except that where the Intramammary Infusion is labeled as sterile, the individual syringes or cartons are sealed and tamper-proof so that sterility is assured at time of use. LABELING: Label it to indicate that it is for veterinary use only. Intramammary Infusion that is sterile may be so labeled. USP REFERENCE STANDARDS 11 USP Cloxacillin Benzathine RS USP Cloxacillin Sodium RS
Cloxacillin Sodium
(Comment on this Monograph)id=m19010=Cloxacillin Sodium=Chl-Cy-Monos.pdf)
C19H17ClN3NaO5S H2O 475.88 4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[[[3-(2chlorophenyl)-5-methyl-4-isoxazolyl]carbonyl]amino]-3,3dimethyl-7-oxo-, monosodium salt, monohydrate, [2S(2,5,6)]-; Monosodium (2S,5R,6R)-6-[3-(o-chlorophenyl)-5-methyl-4isoxazolecarboxamido]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0]heptane-2-carboxylate monohydrate [7081-44-9]. Anhydrous 457.87 [642-78-4]. DEFINITION Cloxacillin Sodium contains the equivalent of NLT 825 g of cloxacillin (C19H18ClN3O5S) per mg. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Sodium 191 ASSAY PROCEDURE Solution A: 0.02 M solution of monobasic potassium phosphate Adjust with 2 N sodium hydroxide to a pH of 6.8.
SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 4.57.5, in 10 mg/mL solution STERILITY TESTS 71: Where the label states that Cloxacillin Sodium is sterile, it meets the requirements when tested as directed for Direct Inoculation of the Culture Medium under Test for Sterility of the Product to Be Examined, except to use Fluid Thioglycollate Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, to use SoybeanCasein Digest Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, and to shake the tubes once daily. WATER DETERMINATION, Method I 921: 3.0%5.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at a temperature not exceeding 25. LABELING: Where it is intended for use in preparing sterile dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of sterile dosage forms. USP REFERENCE STANDARDS 11 USP Cloxacillin Sodium RS
USP 32
Official Monographs / Cloxacillin 159

UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements NMT 5.0%
Cloxacillin Sodium Capsules

(Comment on this Monograph)id=m19020=Cloxacillin Sodium Capsules=Chl-Cy-Monos.pdf) DEFINITION Cloxacillin Sodium Capsules contain the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cloxacillin (C19H18ClN3O5S). ASSAY PROCEDURE Solution A: Prepare a 0.02-M solution of monobasic potassium phosphate in water, and adjust with 2 N sodium hydroxide to a pH of 6.8. Mobile phase: Acetonitrile and Solution A (2:8) Standard solution: 0.55 mg/mL of USP Cloxacillin Sodium RS in Solution A Sample solution: Transfer an equivalent to 100 mg of cloxacillin, from finely powdered Tablets (NLT 10), to a 200mL volumetric flask, dilute with Solution A to volume, and stir for 10 min. Filter 25 mL of this mixture, discarding the first 5 mL of the filtrate. Use the clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 225 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.8 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C19H18ClN3O5S in the portion of Capsule contents taken: Result = (rU/rS) (CS/CU) 100 = peak response of the cloxacillin from the Sample solution = peak response of the cloxacillin from the rS Standard solution = concentration of USP Cloxacillin sodium RS in CS the Standard solution (g/mL) = nominal concentration of cloxacillin in the CU Sample solution (g/mL) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.05 M pH 6.8 potassium phosphate buffer; 900 mL Apparatus 1: 100 rpm Time: 30 min Analysis: Determine the amounts of C19H18ClN3O5S dissolved, by employing the Analysis in the Assay, in comparison with a Standard solution having a known concentration of USP Cloxacillin Sodium RS in the Dissolution Medium. Tolerances: NLT 80% (Q) of the labeled amount of C19H18ClN3O5S rU
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cloxacillin Sodium RS
Cloxacillin Sodium Intramammary Infusion

(Comment on this Monograph)id=m19024=Cloxacillin Sodium Intramammary Infusion=Chl-Cy-Monos.pdf) DEFINITION Cloxacillin Sodium Intramammary Infusion is a suspension of Cloxacillin Sodium in a suitable natural or chemically modified vegetable oil vehicle with a suitable dispersing agent. It has a potency equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of cloxacillin (C19H18ClN3O5S). IDENTIFICATION INFRARED ABSORPTION 197K Sample: Transfer 500 mg of cloxacillin from Intramammary Infusion to a 50-mL centrifuge tube, add 15 mL of isooctane, mix, and centrifuge. Decant and discard the isooctane. Wash the residue with two 15-mL portions of isooctane and two 15-mL portions of ethyl ether, and discard the washings. Dry the residue in a current of air. ASSAY ANTIBIOTICSMICROBIAL ASSAYS 81 Sample: Expel the contents of 1 syringe of Intramammary Infusion into a high-speed glass blender jar containing 499.0 mL of Buffer No. 1 and 1.0 mL of polysorbate 80, and blend for 35 min. Allow to stand for 10 min, and dilute a measured volume of the aqueous phase with Buffer No. 1 to obtain a sample dilution having a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed for cloxacillin. Acceptance criteria: 90.0%120.0% SPECIFIC TESTS STERILITY TESTS 71 Where the label states that it is sterile, it meets the requirements when tested as directed for Direct Inoculation of the Culture Medium under Test for Sterility of the Product to be Examined, except to use Fluid Thioglycollate Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, to use SoybeanCasein Digest Medium containing polysorbate 80 solution (1 in 200) and an amount of sterile penicillinase sufficient to inactivate the cloxacillin in each tube, and to shake the tubes once daily. WATER DETERMINATION, Method I 921: NMT 1.0%, 20 mL of a mixture of toluene and methanol (7:3) being used in place of methanol in the titration vessel ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in disposable syringes that are well-closed containers, except that where the Intramammary Infusion is labeled as sterile, the individual
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USP 32
SPECIFIC TESTS PH 791: 5.07.5, in the solution constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cloxacillin Sodium RS
syringes or cartons are sealed and tamper-proof so that sterility is assured at time of use. LABELING: Label it to indicate that it is for veterinary use only. Intramammary Infusion that is sterile may be so labeled. USP REFERENCE STANDARDS 11 USP Cloxacillin Sodium RS
Cloxacillin Sodium for Oral Solution

(Comment on this Monograph)id=m19030=Cloxacillin Sodium for Oral Solution=Chl-Cy-Monos.pdf) DEFINITION Cloxacillin Sodium for Oral Solution is a dry mixture of Cloxacillin Sodium and one or more suitable buffers, colors, flavors, and preservatives. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of cloxacillin (C19H18ClN3O5S). ASSAY PROCEDURE Solution A: Prepare a 0.02 M solution of monobasic potassium phosphate in water, and adjust with 2 N sodium hydroxide to a pH of 6.8 Mobile phase: Acetonitrile and Solution A (2:8) Standard solution: 0.55 mg/mL of USP Cloxacillin Sodium RS in Solution A Sample solution: Constitute Cloxacillin Sodium for Oral Solution as directed in the labeling. Transfer a measured volume of the resulting solution, equivalent to 125 mg of cloxacillin, to a 250-mL volumetric flask, dilute with Solution A to volume, mix, and stir for 15 min. Filter about 25 mL of this mixture, discarding the first 5 mL of the filtrate. Use the clear filtrate as the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 225-nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.8 Relative standard deviation: NMT 2.0% Analysis Sample: Standard solution and Sample solution Calculate the percentage of C19H18ClN3O5S in the Cloxacillin Sodium for Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response for cloxacillin from the Sample solution = peak response for cloxacillin from the Standard rS solution = concentration of USP Cloxacillin Sodium RS in CS the Standard solution (g/mL) = nominal concentration of the Sample solution CU (g/mL) Acceptance criteria: 90.0%120.0% rU PERFORMANCE TESTS DELIVERABLE VOLUME 698: Meets the requirements UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for solids packaged in single-unit containers
Clozapine
(Comment on this Monograph)id=m19045=Clozapine=Chl-CyMonos.pdf)
326.83 C18H19ClN4 5H-Dibenzo[b,e][1,4]diazepine, 8-chloro-11-(4-methyl-1piperazinyl)-; 8-Chloro-11-(4-methyl-1-piperazinyl)-5H-dibenzo[b,e] [1,4]diazepine [5786-21-0]. DEFINITION Clozapine contains NLT 98.0% and NMT 102.0% of C18H19ClN4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the System suitability solution, as obtained in the Procedure in Organic Impurities. ASSAY PROCEDURE Sample solution: 115 mg of Clozapine in 70 mL of glacial acetic acid Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 16.34 mg of C18H19ClN4. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Diluent: Methanol and water (8:2) Solution A: 2 mg/mL of monobasic potassium phosphate Adjust to a pH of 2.4 with phosphoric acid (85%). [NOTEThe pH of this solution must not be below 2.4.] Solution B: Filtered and degassed mixture of acetonitrile, methanol, and Solution A (1:1:8) Solution C: Filtered and degassed mixture of acetonitrile, methanol, and Solution A (2:2:1) Mobile phase: Use variable mixtures of Solution B and Solution C as directed in the gradient table.
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Time (min) 0-4 4-24 24-29 29-40 Solution B (%) 100 1000 0 0100 Solution C (%) 0 0100 100 1000
Official Monographs / Clozapine 161

Acceptance Criteria Individual impurities: See Impurity Table 1. Total impurities: NMT 0.6%
Impurity Table 1 Approximate Relative Retention Time 0.9 1.0 1.1 1.6 1.7 Relative Response Factor 1.0 0.35 1.2 1.0 1.0 Acceptance Criteria, NMT (%) 0.3 0.2 0.1 0.2 0.10
Elution isocratic linear gradient isocratic linear gradient
System suitability solution: Dissolve 4 mg of USP Resolution Solution Mixture RS in 4 mL of methanol, add 1 mL of water, and dilute with Diluent to 10 mL. Standard stock solution: 0.75 mg/mL of USP Clozapine RS in Diluent Standard solution: 0.75 g/mL of clozapine from the Standard stock solution Sample solution: Dissolve 75.0 mg of the substance to be examined in 80 mL of methanol, and dilute with water to 100.0 mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 257 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution Suitability requirements [NOTELocate the clozapine peak and the peaks due to the specified impurities using the relative retention times provided in Impurity Table 1.] Resolution: NLT 2.5 between Impurity C and clozapine, System suitability solution Relative standard deviation: NMT 5.0% for clozapine, Standard solution Analysis Samples: Standard solution and Sample solution [NOTEDisregard any peak with an area less than 0.5 times the area of the clozapine peak from the Standard solution.] Calculate the percentage of each related compound and any unknown impurity in the portion of Clozapine taken: Result = (rU/rS) (CS/CU) (1/RRF) 100 rU rS CS CU RRF = peak response for any impurity in the Sample solution = peak response of clozapine in the Standard solution = concentration of USP Clozapine RS in the Standard solution (mg/mL) = concentration of Clozapine in the Sample solution (mg/mL) = relative response factor of the impurity according to Impurity Table 1
Impurity Peak Impurity C1 Clozapine Impurity D2 Impurity A3 Impurity B4 Individual unspecified impurity
1 2
8-Chloro-11-(piperazin-1-yl)-5H-dibenzo[b,e][1,4]diazepine. 4-Chloro-N1-(2-((4-methylpiperazin-1-yl)carbonyl)phenyl)benzene-1,2diamine. 38-Chloro-5, 10-dihydro-11H-dibenzo[b,e][1,4]diazepin-11-one. 411,11-(Piperazine-1,4-diyl)bis(8-chloro-5H-dibenzo[b,e][1,4]diazepine).
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 182186 LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clozapine RS USP Clozapine Resolution Mixture RS
Clozapine Tablets
(Comment on this Monograph)id=m19050=Clozapine Tablets=Chl-Cy-Monos.pdf) DEFINITION Clozapine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of clozapine (C18H19ClN4). IDENTIFICATION A. The RF value of the principal spot observed in the chromatogram of the Sample solution corresponds to that of the principal spots observed in the chromatograms of the Standard solutions, as obtained in the Procedure for Organic Impurities.
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suitably diluted with Medium, if necessary, in comparison with the Standard solution. Tolerances: NLT 85% (Q) of the labeled amount of C18H19ClN4. IMPURITIES Organic Impurities PROCEDURE Diluent: Chloroform and methanol (4:1) Standard stock solution: 5.0 mg/mL of USP Clozapine RS in Diluent Standard solutions: Dilute portions of the Standard stock solution with Diluent to obtain the following solutions.
Percentage (%, for comparison with Sample specimen) 0.5 0.4 0.3 0.2 0.1
B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Methanol, triethylamine, and water (800:0.75:200) System suitability stock solution: Transfer 10 mg of clozapine to a suitable container, add 5 mL of 0.1 N hydrochloric acid, and heat for 2 h at 90. Transfer this solution to a 100-mL volumetric flask, add 15 mL of water, and dilute with methanol to volume. System suitability solution: System suitability stock solution and Standard solution (1:1) Standard solution: 0.125 mg/mL of USP Clozapine RS [NOTEDissolve the Reference Standard in methanol and dilute with water to obtain the final concentration. The final solvent composition of methanol and water is about 8:2.] Sample solution: Transfer 125 mg of clozapine, from a quantity of finely powdered Tablets (NLT 20), to a 1-L volumetric flask, dissolve in 640 mL of methanol, sonicate for 10 min, dilute with water to volume, mix, and filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 257 nm Column: 4.0-mm 25-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.5 between the clozapine peak and any other peak, System suitability solution Column efficiency: NLT 1500 theoretical plates, Standard solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H19ClN4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Clozapine RS in the Standard solution (mg/mL) = nominal concentration of clozapine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Solution A: Transfer 2 g of sodium hydroxide to a 1-L volumetric flask, dissolve in 450 mL of water, adjust with glacial acetic acid to a pH of 4.0, and dilute with water to volume. Medium: Solution A, 900 mL Apparatus 1: 100 rpm Time: 45 min Standard solution: USP Clozapine RS in Medium Sample solution: Sample per Dissolution 711 Spectrometric conditions Mode: UV Analytical wavelength: 290 nm Analysis Samples: Standard solution, and Sample solution Determine the amount of C18H19ClN4 dissolved from the UV absorption of filtered portions of the Sample solution,
Standard solution A B C D E
Dilution 1 in 200 1 in 250 1 in 333 1 in 500 1 in 1000
Concentration (g/mL of RS) 25 20 15 10 5
Sample solution: Transfer an equivalent to 125 mg of clozapine, from a portion of finely powdered Tablets (NLT 20), to a 25-mL volumetric flask, dissolve in 20 mL of Diluent, shake by mechanical means for 15 min, dilute with Diluent to volume, and filter. Developing solvent system: n-Heptane, chloroform, dehydrated alcohol, and ammonium hydroxide (30:30:30:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Analysis Samples: Standard solution and Sample solution Proceed as directed under Chromatography 621, ThinLayer Chromatography. Examine the plate under shortwavelength UV light, and compare the intensities of any seondary spots observed in the chromatogram of the Sample solution with those of the principal spots in the chromatograms of the Standard solutions. Acceptance criteria: No secondary spot from the chromatogram of the Sample solution is larger or more intense than the principal spot from Standard solution A (NMT 0.5%), and the sum of the intensities of the secondary spots from the Sample solution corresponds to NMT 2.0%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Clozapine RS
Coal Tar
(Comment on this Monograph)id=m19080=Coal Tar=Chl-CyMonos.pdf) DEFINITION Coal Tar is the tar obtained as a by-product during the destructive distillation of bituminous coal at temperatures in the range of 900 to 1100. It may be processed further either by extraction with alcohol and suitable dispersing agents and
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maceration times or by fractional distillation with or without the use of suitable organic solvents. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
Official Monographs / Cobalt 163

(57Co) in the molecular structure. Each Capsule contains NLT 90.0% and NMT 110.0% of the labeled amount of 57Co as cyanocobalamin expressed in megabecquerels (microcuries) at the time indicated in the labeling. The cyanocobalamin content is NLT 90.0% and NMT 110.0% of the labeled amount. IDENTIFICATION RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is identical to that of a specimen of 57Co of known purity that exhibits a major photopeak having an energy of 0.122 MeV (see Radioactivity 821). ASSAY RADIOACTIVITY (See Radioactivity, 821 Selection of a Counting Assembly .) Analysis: Using a suitable counting assembly, determine the radioactivity, in MBq (Ci)/Capsule, of Cyanocobalamin Co 57 Capsules by use of a calibrated system. Acceptance criteria: 90.0%110.0% of the labeled amount of 57Co as cyanocobalamin CONTENT OF CYANOCOBALAMIN Analysis: Determine the content, in g/Capsule, of cyanocobalamin as directed under Vitamin B12 Activity Assay 171. Acceptance criteria: 90.0%110.0% of the labeled amount of cyanocobalamin PERFORMANCE TESTS DISINTEGRATION 701 Time: 30 min Analysis: Test 1 Capsule in 1 N hydrochloric acid maintained at 37 2 as the immersion fluid UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Content Uniformity: Determine the instrument response of each of 10 Capsules by measurement in a suitable counting assembly and under identical geometric conditions. Calculate the average radioactivity per Capsule. The radioactivity of none of the Capsules differs by more than 10% from the average. The relative standard deviation is less than 3.5%. SPECIFIC TESTS RADIOCHEMICAL PURITY Solution A: 10 mg/mL of dibasic sodium phosphate and adjust with phosphoric acid to a pH of 3.5 Mobile phase: Methanol and Solution A (26.5:73.5) [NOTEUse within 2 days.] Standard solution: 2 g/mL of cyanocobalamin in Mobile phase Sample solution: Dissolve the contents of 1 Capsule in 1 mL of water, allow to stand for 10 min, and centrifuge. Use the supernatant. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 361 nm, and gamma detector adjusted for 57Co Column: 4.6-mm 25-cm; stainless steel column; 5-m packing L7 Flow rate: 1 mL/min Injection size: 100 L Analysis Samples: Standard solution and Sample solution Inject the Standard solution into the chromatograph, record the chromatogram for 30 min, and note the retention time of the cyanocobalamin peak. Inject the Sample solution into the chromatograph, and record the chromatogram for three times the retention time of cyanocobalamin. Measure the peak areas using the gamma detector.
NMT 2.0%, from 100 mg
Coal Tar Ointment

(Comment on this Monograph)id=m19130=Coal Tar Ointment=Chl-Cy-Monos.pdf) DEFINITION Prepare Coal Tar Ointment as follows.
Coal Tar Polysorbate 80 Zinc Oxide Paste To make 10 g 5g 985 g 1000 g
Blend the Coal Tar with the Polysorbate 80, and incorporate the mixture with the Zinc Oxide Paste. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers.
Coal Tar Topical Solution

(Comment on this Monograph)id=m19140=Coal Tar Topical Solution=Chl-Cy-Monos.pdf) DEFINITION Prepare Coal Tar Topical Solution as follows.
Coal Tar Polysorbate 80 Alcohol To make 200 g 50 g A sufficient quantity 1000 mL
Mix the Coal Tar with 500 g of washed sand (see Reagents, Indicators, and SolutionsReagents), and add the Polysorbate 80 and 700 mL of Alcohol. Macerate the mixture for 7 days in a closed vessel with frequent agitation. Filter, and rinse the vessel and the filter with sufficient Alcohol to make the product measure 1000 mL. OTHER COMPONENTS Alcohol Determination 611: 81.0%86.0% of C2H5OH
Cyanocobalamin Co 57 Capsules
(Comment on this Monograph)id=m19200=Cyanocobalamin Co 57 Capsules=Chl-Cy-Monos.pdf) Vitamin B12-57Co; Vitamin B12-57Co [41559-38-0; 13115-03-2]. DEFINITION Cyanocobalamin Co 57 Capsules contain Cyanocobalamin in which a portion of the molecules contain radioactive cobalt
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Calculate the percentage of cyanocobalamin present as cyanocobalamin 57Co in the portion of Capsules taken: Result = (rU/rT) 100
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ASSAY RADIOACTIVITY (See Radioactivity, 821 Selection of a Counting Assembly .) Analysis: Using a suitable counting assembly, determine the radioactivity, in MBq (Ci)/mL, of Oral Solution by use of a calibrated system as directed. Acceptance criteria: 90.0%110.0% of the labeled amount of 57Co as cyanocobalamin CONTENT OF CYANOCOBALAMIN Analysis: Determine the content, in g/Capsule, of cyanocobalamin as directed under Vitamin B12 Activity Assay 171. Acceptance criteria: 90.0%110.0% of the labeled amount of cyanocobalamin PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for oral solution packaged in single-unit containers DELIVERABLE VOLUME 698: Meets the requirements for oral solution packaged in multiple-unit containers SPECIFIC TESTS PH 791: 4.05.5 RADIOCHEMICAL PURITY Solution A: 10 mg/mL of dibasic sodium phosphate Adjust with phosphoric acid to a pH of 3.5. Mobile phase: Methanol and Solution A (26.5:73.5) [NOTEUse within 2 days.] Standard solution: 2 g/mL of cyanocobalamin in Mobile phase Sample solution: Oral Solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 361 nm, and gamma detector adjusted for 57Co Column: 4.6-mm 25-cm; stainless steel column; 5-m packing L7 Flow rate: 1 mL/min Injection size: 100 L Analysis Samples: Standard solution and Sample solution Inject the Standard solution into the chromatograph, record the chromatogram for 30 min, and note the retention time of the cyanocobalamin peak. Inject the Sample solution into the chromatograph, and record the chromatogram for three times the retention time of cyanocobalamin. Measure the peak areas using the gamma detector. Calculate the percentage of cyanocobalamin present as cyanocobalamin 57Co in the portion of Oral Solution taken: Result = (rU/rT) 100 = peak area for cyanocobalamin 57Co of the Sample solution = sum of all the peak areas in the rT radiochromatogram of the Sample solution Acceptance criteria: NLT 90% of the total radioactivity is found as cyanocobalamin 57Co. RADIONUCLIDIC PURITY Analysis: Using a suitable calibrated instrument (see Radioactivity 821) and standardized solutions of 58Co, 57Co, and 60Co, record the gamma spectrum of the Oral Solution. The spectrum does not differ significantly from that of the standardized 57Co solution. Determine the relative amounts of 58Co, 57Co, and 60Co present. Cobalt 58 has a half-life of 70.9 days, and its presence is shown by 0.511-MeV and 0.811-MeV gamma photons. Cobalt 60 has a half-life of 5.27 years, and its presence is shown by 1.173-MeV and 1.333-MeV gamma photons. rU
= peak area for cyanocobalamin 57Co of the Sample solution = total of all the peak areas in the rT radiochromatogram of the Sample solution Acceptance criteria: NLT 90% of the total radioactivity is found as cyanocobalamin 57Co. RADIONUCLIDIC PURITY Sample solution: Dissolve the contents of 1 Capsule in 1 mL of water, allow to stand for 10 min, and centrifuge. Use the supernatant. Analysis: Using a suitable calibrated instrument (see Radioactivity 821) and standardized solutions of 58Co, 57Co, and 60Co, record the gamma spectrum of the Sample Solution. The spectrum does not differ significantly from that of the standardized 57Co solution. Determine the relative amounts of 58Co, 57Co, and 60Co present. Cobalt 58 has a half-life of 70.9 days, and its presence is shown by 0.511MeV and 0.811-MeV gamma photons. Cobalt 60 has a halflife of 5.27 years, and its presence is shown by 1.173-MeV and 1.333-MeV gamma photons. Acceptance criteria: NMT 1% of the total radioactivity is due to 60Co; and NMT 2% of the total radioactivity is due to 58Co, 60Co, and other radionuclidic impurities. SPECIFIC ACTIVITY: NLT 0.02 MBq (0.5 Ci)/g of cyanocobalamin rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers, and store in a cold place. LABELING: Label the Capsules to include the following: the date of calibration; the amount of cyanocobalamin expressed in g/Capsule; the amount of 57Co as cyanocobalamin expressed in megabecquerels (microcuries)/Capsule at the time of calibration; the expiration date; and the statement [CAUTIONRadioactive Material]. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay, and also indicates that the radioactive half-life of 57Co is 270.9 days. USP REFERENCE STANDARDS 11 USP Cyanocobalamin RS
Cyanocobalamin Co 57 Oral Solution

(Comment on this Monograph)id=m19210=Cyanocobalamin Co 57 Oral Solution=Chl-Cy-Monos.pdf) Vitamin B12-57Co; Vitamin B12-57Co [41559-38-0; 13115-03-2]. DEFINITION Cyanocobalamin Co 57 Oral Solution is a solution suitable for oral administration, containing Cyanocobalamin in which a portion of the molecules contain radioactive cobalt (57Co) in the molecular structure. Cyanocobalamin Co 57 Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of 57Co as cyanocobalamin expressed in megabecquerels (microcuries) per mL at the time indicated in the labeling. The cyanocobalamin content is NLT 90.0% and NMT 110.0% of the labeled amount. Cyanocobalamin Co 57 Oral Solution contains a suitable antimicrobial agent. IDENTIFICATION RADIONUCLIDIC IDENTIFICATION: Its gamma-ray spectrum is identical to that of a specimen of 57Co of known purity that exhibits a major photopeak having an energy of 0.122 MeV (see Radioactivity 821).
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Acceptance criteria: NMT 1% of the total radioactivity is due to 60Co; and NMT 2% of the total radioactivity is due to 58Co, 60Co, and other radionuclidic impurities. SPECIFIC ACTIVITY: NLT 0.02 MBq (0.5 Ci/g of cyanocobalamin ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, and store in a cold place. LABELING: Label it to include the following: the date of calibration; the amount of 57Co as cyanocobalamin expressed as total megabecquerels (microcuries) and as megabecquerels (microcuries) per mL at the time of calibration; the amount of cyanocobalamin expressed in g/mL; the name and quantity of the added preservative; the expiration date; and the statement [CAUTION Radioactive Material]. The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay, and also indicates that the radioactive half-life of 57Co is 270.9 days, and directs that the Oral Solution be protected from light. USP REFERENCE STANDARDS 11 USP Cyanocobalamin RS
Official Monographs / Cobalt 165

UNIFORMITY OF DOSAGE UNITS, Procedure for Content Uniformity 905 Analysis: Determine the instrument response of each of 10 Capsules by measurement in a suitable counting assembly and under identical geometric conditions. Calculate the average radioactivity per Capsule. The radioactivities of none of the Capsules differ by more than 10% from the average. The relative standard deviation is less than 3.5%. Acceptance criteria: Meet the requirements SPECIFIC TESTS RADIOCHEMICAL PURITY Solution A: 10 mg/mL of dibasic sodium phosphate Adjust with phosphoric acid to a pH of 3.5. Mobile phase: Methanol and Solution A (26.5:73.5) [NOTEUse within 2 days.] Standard solution: 2 g/mL of cyanocobalamin in Mobile phase Sample solution: Dissolve the contents of 1 Capsule in 1 mL of water, allow to stand for 10 min, and centrifuge. Use the supernatant. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 361 nm, and gamma detector adjusted for 58Co Column: 4.6-mm 25-cm; stainless steel column; 5 m packing L7 Flow rate: 1 mL/min Injection size: 100 L Analysis Samples: Standard solution and Sample solution Inject the Standard solution into the chromatograph, record the chromatogram for 30 min, and note the retention time of the cyanocobalamin peak. Inject the Sample solution into the chromatograph, and record the chromatogram for three times the retention time of cyanocobalamin. Measure the peak areas using the gamma detector. Calculate the percentage of cyanocobalamin present as cyanocobalamin 58Co in the portion of Capsules taken: Result = (rU/rT) 100 = peak response for cyanocobalamin 58Co from the Sample solution = sum of all peak responses in the rT radiochromatogram from the Sample solution Acceptance criteria: NLT 90% of the total radioactivity is found as cyanocobalamin 58Co RADIONUCLIDIC PURITY Analysis: Using a suitable calibrated instrument (see Radioactivity 821) and standardized solutions of 58Co, 57Co, and 60Co, record the gamma spectrum. The spectrum does not differ significantly from that of the standardized 58Co solution. Determine the relative amounts of 58Co, 57Co, and 60Co present. Cobalt 57 has a half-life of 270.9 days, and its presence is shown by 0.122 MeV gamma photons. Cobalt 60 has a half-life of 5.27 years and its presence is shown by 1.173 MeV and 1.333 MeV gamma photons. Acceptance criteria: NMT 1% of the total radioactivity is due to 60Co; and NMT 2% of the total radioactivity is due to 57Co, 60Co, and other radionuclidic impurities. Specific Activity: NLT 0.02 MBq (or 0.5 Ci) per g of cyanocobalamin. rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers, and store in a cold place. LABELING: Label it to include the following: the date of calibration; the amount of cyanocobalamin expressed in
Cyanocobalamin Co 58 Capsules
(Comment on this Monograph)id=m19215=Cyanocobalamin Co 58 Capsules=Chl-Cy-Monos.pdf) Vitamin B1258
Co
DEFINITION Cyanocobalamin Co 58 Capsules contain Cyanocobalamin in which a portion of the molecules contain radioactive cobalt (58Co) in the molecular structure. Each Capsule contains NLT 90.0% and NMT 110.0% of the labeled amount of 58Co as cyanocobalamin expressed in megabecquerels (or microcuries) at the time indicated in the labeling. The cyanocobalamin content is NLT 90.0% and NMT 110.0% of the labeled amount. IDENTIFICATION RADIOACTIVITY, Identification and Assay of Radionuclides 821 A. Its gamma-ray spectrum is identical to that of a specimen of 58Co that exhibits major photopeaks at 0.511 MeV (annihilation radiation) and 0.811 MeV. B. The retention time of the major peak in the radiochromatogram of the Sample solution corresponds to that in the chromatogram of the Standard solution, as obtained in the test for Radiochemical Purity. ASSAY RADIOACTIVITY Analysis: Using a suitable counting assembly (see Radioactivity, Selection of a Counting Assembly 821), and calibrated system, determine the radioactivity, in MBq (or Ci)/Capsule, of Cyanocobalamin Co 58 Capsules. Acceptance criteria: 90.0%110.0% of the labeled amount of 58Co as cyanocobalamin CONTENT OF CYANOCOBALAMIN Analysis: Determine the content, in g/Capsule, of cyanocobalamin as directed under Vitamin B12 Activity Assay 171. Acceptance criteria: 90.0%110.0% of the labeled amount of cyanocobalamin PERFORMANCE TESTS DISINTEGRATION 701 Time: 30 min Analysis: Test 1 Capsule in 1 N hydrochloric acid maintained at 37 2 as the immersion fluid.
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determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 30.34 mg of C17H21NO4. Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities LIMIT OF CINNAMYL-COCAINE AND OTHER REDUCING SUBSTANCES Sample solution: Dissolve 300 mg of finely powdered Cocaine in 1 mL of dilute hydrochloric acid (1 in 12) with the aid of heat, if necessary, and dilute with water to 15 mL. Analysis: Mix 5 mL of the Sample solution with 0.3 mL of dilute sulfuric acid (1 in 35) and 0.1 mL of potassium permanganate solution (1 in 300). Acceptance criteria: The violet color does not disappear entirely within 30 min. LIMIT OF ISOATROPYL-COCAINE Sample: 5 mL of the Sample solution prepared in the test for Limit of Cinnamyl-Cocaine and Other Reducing Substances Analysis: Dilute the Sample with 80 mL of water, add 0.2 mL of 6 N ammonium hydroxide, and stir the solution vigorously for 5 min, occasionally rubbing the inner wall of the beaker with a stirring rod. Acceptance criteria: A crystalline precipitate of cocaine is formed, and the supernatant is clear. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 9698 LOSS ON DRYING 731: Dry it over phosphorus pentoxide for 3 h: it loses NMT 1.0% of its weight. READILY CARBONIZABLE SUBSTANCES TEST 271 Sample solution: Dissolve 500 mg in 5 mL of sulfuric acid. TS Acceptance criteria: The solution has no more color than Matching Fluid A. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Cocaine Hydrochloride RS
g/Capsule; the amount of 58Co as cyanocobalamin expressed in MBq (or Ci)/Capsule at the time of calibration; the expiration date; and the statement [Caution Radioactive Material.] The labeling indicates that in making dosage calculations, correction is to be made for radioactive decay, and also indicates that the radioactive half-life of 58Co is 70.9 days. USP REFERENCE STANDARDS 11 USP Cyanocobalamin RS
Cocaine
(Comment on this Monograph)id=m19300=Cocaine=Chl-CyMonos.pdf)
C17H21NO4 303.35 8-Azabicyclo[3.2.1]octane-2-carboxylic acid, 3-(benzoyloxy)-8methyl-, methyl ester, [1R-(exo,exo)]-; Methyl 3-hydroxy-1h,5h-tropane-2-carboxylate benzoate (ester) [50-36-2]. DEFINITION Cocaine, dried over phosphorus pentoxide for 3 h, contains NLT 99.0% and NMT 101.0% of C17H21NO4. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: 15 g/mL in dilute hydrochloric acid (1 in 120) Acceptance criteria: Absorptivities at 233 nm, calculated on the dried basis, do not differ by more than 3.0%. B. IDENTIFICATIONORGANIC NITROGENOUS BASES 181: Meets the requirements, USP Cocaine Hydrochloride RS being used, and sodium carbonate TS being used in place of sodium hydroxide TS. C. PROCEDURE Sample: 100 mg Analysis 1: Dissolve the Sample in a mixture of 0.4 mL of dilute hydrochloric acid (1 in 12) and water to make 5 mL, and add 5 drops of chromium trioxide solution (1 in 20). Acceptance criteria 1: A yellow precipitate is formed, and it quickly redissolves when the mixture is shaken gently. Analysis 2: Add 1 mL of hydrochloric acid to the solution obtained from Analysis 1. Acceptance criteria 2: A permanent, orange-colored, crystalline precipitate is formed. D. PROCEDURE Sample: 10 mg Analysis: Dissolve the Sample in 1 mL of dilute hydrochloric acid (1 in 600), and evaporate on a steam bath just to dryness. Dissolve the residue in 2 drops of water, and add 1 mL of potassium permanganate solution (1 in 300). Acceptance criteria: A violet, crystalline precipitate is formed, and it appears brownish violet when collected on a filter, and shows characteristic violet-red crystalline aggregates under the low power of a microscope, similar to those obtained from USP Cocaine Hydrochloride RS. ASSAY PROCEDURE Sample: 600 mg, previously dried Analysis: Dissolve the Sample in 50 mL of glacial acetic acid, and add 1 drop of crystal violet TS. Titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank
Cocaine Hydrochloride
(Comment on this Monograph)id=m19330=Cocaine Hydrochloride=Chl-Cy-Monos.pdf) 339.81 C17H21NO4 HCl 8-Azabicyclo[3.2.1]octane-2-carboxylic acid, 3-(benzoyloxy)-8methyl-, methyl ester, hydrochloride, 1R-(exo,exo)-; Methyl 3-hydroxy-1h,5h-tropan-2-carboxylate, benzoate (ester) hydrochloride [53-21-4]. DEFINITION Cocaine Hydrochloride contains NLT 99.0% and NMT 101.0% of C17H21NO4 HCl, calculated on the dried basis. IDENTIFICATION A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181: Meets the requirements, sodium carbonate TS being used in place of 1 N sodium hydroxide B. PROCEDURE Sample solution: 1 in 50 Analysis 1: To 5 mL of the Sample solution, add 5 drops of 50 mg/mL chromium trioxide solution. Acceptance criteria 1: A yellow precipitate is formed, and it quickly redissolves when the mixture is shaken gently. Analysis 2: Add 1 mL of hydrochloric acid to the solution obtained from Analysis 1.
USP 32
Acceptance criteria 2: A permanent, orange-colored crystalline precipitate is formed. C. PROCEDURE Sample solution: 10 mg of Cocaine Hydrochloride in 2 drops of water Analysis: To the Sample solution, add 1 mL of 0.1 N potassium permanganate. Acceptance criteria: A violet, crystalline precipitate is formed, and it appears brownish violet when collected on a filter, and shows characteristic, violet-red crystalline aggregates under the low power of a microscope. D. IDENTIFICATION TESTSGENERAL, Chloride 191: Meets the requirements of the tests ASSAY PROCEDURE Sample solution: Dissolve 500 mg of Cocaine Hydrochloride in a mixture of 40 mL of glacial acetic acid and 10 mL of mercuric acetate TS. Analysis: To the Sample solution, add 2 drops of quinaldine red TS and titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 33.98 mg of C17H21NO4 HCl. Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities LIMIT OF CINNAMYL-COCAINE AND OTHER REDUCING SUBSTANCES Sample solution: 1 in 50 Analysis: To 5 mL of the Sample solution, add 0.3 mL of 1 N sulfuric acid and 0.10 mL of 0.10 N potassium permanganate. Acceptance criteria: The violet color does not disappear entirely within 30 min. LIMIT OF ISOATROPYL-COCAINE Sample solution: 1 in 50 Analysis: Dilute 5 mL of the Sample solution in a beaker with 80 mL of water, add 0.2 mL of 6 N ammonium hydroxide, stir the solution vigorously during 5 min, occasionally rubbing the inner wall of the beaker with a stirring rod. Acceptance criteria: A crystalline precipitate of cocaine is formed, and the supernatant is clear. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S Sample solution: 20 mg/mL, previously dried, in water Acceptance criteria: 71 to 73 ACIDITY Analysis: Dissolve 500 mg in 10 mL of water, add 1 drop of methyl red TS, and titrate with 0.020 N sodium hydroxide. Acceptance criteria: NMT 0.50 mL is required to produce a yellow color. LOSS ON DRYING 731: Dry it over silica gel for 3 h: It loses NMT 1.0% of its weight. READILY CARBONIZABLE SUBSTANCES TEST 271 Sample solution: Dissolve 500 mg in 5 mL of sulfuric acid TS. Acceptance criteria: The Sample solution has no more color than Matching Fluid F. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers.
Official Monographs / Cocaine 167

USP REFERENCE STANDARDS 11 USP Cocaine Hydrochloride RS
Cocaine Hydrochloride Tablets for Topical Solution

(Comment on this Monograph)id=m19360=Cocaine Hydrochloride Tablets for Topical Solution=Chl-Cy-Monos.pdf) DEFINITION Cocaine Hydrochloride Tablets for Topical Solution contain NLT 91.0% and NMT 109.0% of the labeled amount of C17H21NO4 HCl. IDENTIFICATION A. PROCEDURE Sample solution: Equivalent to cocaine hydrochloride (1 in 50) from an appropriately diluted quantity of Tablets Filter the solution. Analysis 1: To 5 mL of the Sample solution, add 5 drops of 50 mg/mL chromium trioxide solution. Acceptance criteria 1: A yellow precipitate is formed, and it redissolves when the mixture is shaken gently. Analysis 2: Add 1 mL of hydrochloric acid to the solution obtained from Analysis 1. Acceptance criteria 2: A permanent, yellowish orange, crystalline precipitate is formed. B. PROCEDURE Sample: A quantity of powdered Tablets equivalent to 10 mg of cocaine hydrochloride Analysis: Dissolve the Sample in 1 mL of water, filter, and add 2 mL of 0.1 N potassium permanganate. Acceptance criteria: A red-purple, crystalline precipitate, which appears brown when collected on a filter, is formed, and it shows characteristic, crystalline aggregates under the low power of a microscope. C. PROCEDURE Sample solution: Prepare a solution equivalent to cocaine hydrochloride (1 in 20) from an appropriately diluted quantity of Tablets; filter the solution. Analysis: Add silver nitrate TS, dropwise, to the Sample solution. Acceptance criteria: A white precipitate is formed, and it is insoluble in nitric acid. ASSAY PROCEDURE Sample solution: Dissolve an equivalent to 60 mg of cocaine hydrochloride, from an amount of finely powdered Tablets (NLT 20), in 10 mL of water, render the solution slightly alkaline with 6 N ammonium hydroxide, and completely extract the cocaine with small successive portions of ether. Evaporate the combined ether extracts on a steam bath to one-half their volume, transfer the remaining liquid to a separator, and wash with three 5-mL portions of water. Shake the water washings with a small portion of ether, and add the ether washing to the combined ether extracts. Add 10.0 mL of 0.05 N sulfuric acid VS to the ether solution, agitate the mixture thoroughly, and draw off the acidified water layer into a beaker. Wash the ether with two small portions of water, and add the washings to the acid liquid. Analysis: Titrate the excess acid in the Sample solution with 0.02 N sodium hydroxide VS, using methyl red TS as the indicator. Each mL of 0.05 N sulfuric acid is equivalent to 16.99 mg of C17H21NO4 HCl.
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Cocaine / Official Monographs

91.0%109.0%
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Dissolve the Cocaine and Tetracaine Hydrochlorides in 25 mL of Purified Water, and add the Epinephrine Injection (1:1000). Separately dissolve Edetate Disodium in Sodium Chloride Injection (0.9%), and dilute, if necessary, with Sodium Chloride Injection (0.9%) to obtain 35 mL of a solution containing 6.4 mg of Edetate Disodium. Similarly, and separately, dissolve a quantity of Benzalkonium Chloride in Purified Water (or use a volume of Benzalkonium Chloride Solution), and dilute, if necessary, with Purified Water to obtain 10 mL of a solution containing 10 mg of Benzalkonium Chloride. Combine the three solutions, add sufficient Purified Water to make the preparation measure 100 mL, and mix to produce the Topical Solution. ASSAY TETRACAINE HYDROCHLORIDE Solution A: 6.3 mg/mL of monobasic potassium phosphate and 0.55 mg/mL of sodium 1-octanesulfonate in water Adjust with phosphoric acid to a pH of 2.5. Solution B: Acetonitrile and Solution A (1:9) Pass the resulting solution through a filter having a 5-m or finer porosity, and degas. Solution C: Acetonitrile and Solution A (3:7) Pass the resulting solution through a filter having a 5-m or finer porosity, and degas. Mobile phase: Use variable mixtures of Solution B and Solution C as directed in the gradient table below.
Time (min) 0 5 10 24 25 75 Solution B (%) 100 100 0 0 100 100 Solution C (%) 0 0 100 100 0 0
PERFORMANCE TESTS DISINTEGRATION 701 Time: 15 min UNIFORMITY OF DOSAGE UNITS, Content Uniformity 905 Standard solution: 80 g/mL of USP Cocaine Hydrochloride RS Sample solution: Place 1 Tablet in a 100-mL volumetric flask, add 50 mL of water, and shake the flask until the tablet is dissolved. Dilute with water to volume, mix, and filter, discarding the first 20 mL of the filtrate. Dilute a portion of the filtrate, if necessary, with water to obtain a solution containing 80 g/mL of cocaine hydrochloride. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 275 nm Cell: 1 cm Blank: Water Analysis Samples: Standard solution and Sample solution Calculate the quantity, in mg, of C17H21NO4 HCl in the Tablet: Result = (Cs/CU (AU/AS) L = concentration of USP Cocaine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of cocaine hydrochloride CU in the Sample solution (g/mL) AU = absorbance of the Sample solution AS = absorbance of the Standard solution L = labeled quantity of cocaine hydrochloride in the Tablet (mg) Acceptance criteria: Meets the requirements Cs ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Cocaine Hydrochloride RS
Cocaine and Tetracaine Hydrochlorides and Epinephrine Topical Solution

(Comment on this Monograph)id=m19375=Cocaine and Tetracaine Hydrochlorides and Epinephrine Topical Solution=Chl-Cy-Monos.pdf) DEFINITION Cocaine and Tetracaine Hydrochlorides and Epinephrine Topical Solution contains NLT 3.6 g and NMT 4.4 g of Cocaine Hydrochloride, NLT 0.90 g and NMT 1.10 g of Tetracaine Hydrochloride, and NLT 20 mg and NMT 30 mg of Epinephrine in 100 mL of Topical Solution prepared as follows (see Pharmaceutical CompoundingNonsterile Preparations 795):
Cocaine Hydrochloride Tetracaine Hydrochloride Epinephrine Injection (1:1000) Benzalkonium Chloride Edetate Disodium Sodium Chloride Injection (0.9%) Purified Water, a sufficient quantity to make 100 mL 4.0 g 1.0 g 25.0 mL 10 mg 6.4 mg 35 mL
Standard solution: 0.5 mg/mL of USP Tetracaine Hydrochloride RS Sample solution: Topical Solution and water (0.5:9.5) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 3.9-mm 30-cm; 10 m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of tetracaine hydrochloride C15H24N2O2 HCl in 100 mL of the Topical Solution taken: Result = (rU/rS) 2CS rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Tetracaine Hydrochloride, in the Standard solution (mg/mL)
USP 32
Acceptance criteria: 0.901.10 g/100 mL COCAINE HYDROCHLORIDE Solution A, Solution B, Solution C, Sample solution, and Chromatographic system Proceed as directed in the Assay for Tetracaine Hydrochloride. Standard solution: 2 mg/mL of USP Cocaine Hydrochloride RS Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of cocaine hydrochloride (C17H21NO4 HCl) in 100 mL of the Topical Solution taken: Result = (rU/rS) 2CS = peak response from the Sample solution = peak response from the Standard solution = concentration of the USP Cocaine Hydrochloride RS in the Standard solution (mg/mL) Acceptance criteria: 3.64.4 g/100 mL EPINEPHRINE Solution A, Solution B, Solution C, Sample solution, and Chromatographic system Proceed as directed in the Assay for Tetracaine Hydrochloride. Standard solution: 3 mg of USP Epinephrine Bitartrate RS in 25-mL water. Mix the resultant solution and water (4:21). Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of epinephrine (C9H13NO3) in 100 mL of the Topical Solution taken: rU rS CS Result = (rU/rS) 2CS (Mr1/Mr2) = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Epinephrine Bitartrate RS in the Standard solution (mg/mL) = molecular weight of epinephrine, 183.20 Mr1 = molecular weight of epinephrine bitartrate, Mr2 333.29 Acceptance criteria: 2030 mg/100 mL rU rS CS SPECIFIC TESTS PH 791: 4.06.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in sterile, tight, lightresistant containers. Store in a refrigerator. LABELING: Label it to state that it is intended for external use only and that it is not to be used if a precipitate is present. The label states that it is to be protected from light. BEYOND-USE DATE: 30 days after the date on which it was compounded USP REFERENCE STANDARDS 11 USP Cocaine Hydrochloride RS USP Epinephrine Bitartrate RS USP Tetracaine Hydrochloride RS
Official Monographs / Cod 169

Coccidioides immitis. It has a potency such that the 1:100 dilution is bioequivalent to the U.S. Reference Coccidioidin 1:100. It contains a suitable antimicrobial agent. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve at a temperature between 2 and 8. EXPIRATION DATE: The expiration date is not later than 3 years after the date of issue from the manufacturers cold storage (5, 1 year) for the mycelial product and not later than 18 months after the date of issue from the manufacturers cold storage (5, 18 months) for the spherule-derived product. LABELING: Label it to state that any dilutions made of the product should be stored in a refrigerator and used within 24 h. Label it also to state that a separate syringe and needle shall be used for each individual injection.
Cod Liver Oil

(Comment on this Monograph)id=m19490=Cod Liver Oil=ChlCy-Monos.pdf) DEFINITION Cod Liver Oil is the partially destearinated fixed oil obtained from fresh livers of Gadus morrhua Linn and other species of e Fam. Gadidae. Cod Liver Oil contains, in each g, NLT 180 g (600 USP Units) and NMT 750 g (2500 USP Units) of vitamin A and NLT 1.5 g (60 USP Units) and NMT 6.25 g (250 USP Units) of vitamin D. Cod Liver Oil may be flavored by the addition of NMT 1% of a suitable flavor or a mixture of flavors. A suitable antioxidant may be added. IDENTIFICATION A. PRESENCE OF VITAMIN A Analysis: To 1 mL of a 25 mg/mL solution in chloroform, add 10 mL of antimony trichloride TS. Acceptance criteria: A blue color results immediately. B. FATTY ACID PROFILE Antioxidant solution: 0.05 mg/mL of butylated hydroxytoluene in hexanes Standard solution: 45 g/mL of USP Cod Liver Oil RS in Antioxidant solution Transfer 2.0 mL of the resulting solution into a quartz tube, and evaporate with a gentle stream of nitrogen. Add 1.5 mL of a 2% solution of sodium hydroxide in methanol, cap tightly with a polytetrafluoroethylene-lined cap, mix, and heat in a water bath for 7 min. Cool, add 2 mL of a 120 mg/mL solution of boron trichloride in methanol, cover with nitrogen, cap tightly, mix, and heat in a water bath for 30 min. Cool to 40 to 50, add 1 mL of isooctane, cap, and mix in a vortex mixer or shake vigorously for at least 30 s. Immediately add 5 mL of saturated sodium chloride solution, cover with nitrogen, cap, and mix in a vortex mixer or shake thoroughly for at least 15 s. Allow the upper layer to become clear, and transfer to a separate tube. Shake the methanol layer once more with 1 mL of isooctane, and combine the isooctane extracts. Wash the combined extracts twice with 1 mL of water, and dry over anhydrous sodium sulfate. System suitability solution: Prepare a mixture containing equal amounts of methyl palmitate, methyl stearate, methyl arachidate, and methyl behenate. [NOTEA suitable mixture is available from Supelco, Bellefonte, PA, as GLC-40 cat. number 1895-1AMP].
Coccidioidin
(Comment on this Monograph)id=m19390=Coccidioidin=ChlCy-Monos.pdf) DEFINITION Coccidioidin conforms to the regulations of the FDA concerning biologics (see Biologics 1041). It is a sterile solution containing the antigens obtained from the by-products of mycelial growth or from the spherules of the fungus
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Cod / Official Monographs

Lower limit (area %) 2.0 7.0 1.0 Upper limit (area %) 6.0 14.0 4.0
USP 32
14:0, 15:0, 16:0, 16:1 n-7, 16:4 n-1, 18:0, 18:1 n-9, 18:1 n-7, 18:2 n-6, 18:3 n-3, 18:4 n-3, 20:1 n-11, 20:1 n-9, 20:1 n-7, 20:2 n-6, 20:4 n-6, 20:3 n-3, 20:4 n-3, 20:5 n-3, 22:1 n-11, 22:1 n-9, 21:5 n-3, 22:5 n-3, and 22:6 n-3.) [NOTEThe chromatogram obtained from the Standard solution is similar to the reference chromatogram supplied with USP Cod Liver Oil RS. Identify the retention times of the relevant fatty acid methyl esters by comparing the chromatogram of the Standard solution with the reference chromatogram supplied with USP Cod Liver Oil RS.] Analysis Samples: Standard solution and Sample solution Identify the retention times of the relevant fatty acid methyl esters in the Sample solution by comparing the chromatogram of the Sample solution with that of the Standard solution. Calculate the area percentage for each fatty acid methyl ester taken: Result = (rA/rB) 100 = average peak area of each individual fatty acid = total peak area from all peaks, except the solvent front and butylated hydroxytoluene Acceptance criteria: The fatty acids observed in the Sample solution should meet the limits described in the table above. ASSAY VITAMIN A Sample: Using 500 mg to 1 g of Oil, proceed as directed under Vitamin A Assay 571. Acceptance criteria: 180 g (600 USP Units) 750 g (2500 USP Units) of vitamin A/g VITAMIN D Mobile phase A: n-amyl alcohol and dehydrated hexane (3:997) (See Chromatography 621, System Suitability.) Mobile phase B: Acetonitrile, water, and phosphoric acid (96:3.8:0.2) (See Chromatography 621, System Suitability.) Solution A: 10 mg/mL of butylated hydroxytoluene in chromatographic hexane Solution B: Dissolve 800 mg/mL of potassium hydroxide in freshly boiled water, mix, and cool. [NOTEPrepare this solution fresh daily.] Solution C: 30 mg/mL of potassium hydroxide in freshly boiled water, and alcohol (9:1) [NOTEDissolve in water and then add alcohol. Prepare this solution fresh daily.] Solution D: 100 mg/mL of ascorbic acid in water [NOTEPrepare this solution fresh daily.] Internal standard solution: 5 g/mL of USP Ergocalciferol RS in alcohol Standard solution: 5 g/mL of USP Cholecalciferol RS in ethyl alcohol. Transfer 2.0 mL of this solution and 2.0 mL of the Internal standard solution to a round-bottomed flask. Proceed as directed for the Sample solution A beginning with Add 5 mL of. Sample solution A: Transfer a quantity of 4.00 g of Cod Liver Oil, to a round-bottomed flask. Add 5 mL of Solution D, 100 mL of alcohol, and 10 mL of Solution B, and mix. Reflux the mixture on a steam bath for 30 min. Add 100 mL of 10 mg/mL sodium chloride solution. Cool rapidly under running water, and transfer the saponified mixture to a 500mL separator, rinsing the saponification flask with 75 mL of 10 mg/mL sodium chloride solution, and then with 150 mL of a mixture of ether and hexane (1:1). Shake the combined saponified mixture and rinsings vigorously for 30 s, and rA rB
Fatty acid Saturated fatty acids: Myristic acid Palmitic acid Stearic acid Monounsaturated fatty acids: Palmitoleic acid cis-Vaccenic acid Oleic acid Gadoleic acid Gondoic acid Erucic acid Cetoleic acid Polyunsaturated fatty acids: Linoleic acid -Linolenic acid Moroctic acid Eicosapentaenoic acid Docosahexaenoic acid
Shorthand notation 14:0 16:0 18:0
16:1 n-7 18:1 n-7 18:1 n-9 20:1 n-11 20:1 n-9 22:1 n-9 22:1 n-11
4.5 2.0 12.0 1.0 5.0 0 5.0
11.5 7.0 21.0 5.5 17.0 1.5 12.0
18:2 n-6 18:3 n-3 18:4 n-3 20:5 n-3 22:6 n-3
0.5 0 0.5 7.0 6.0
3.0 2.0 4.5 16.0 18.0
Sample solution: Proceed as directed for the Standard solution, except to use a weighed quantity of Cod Liver Oil. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 0.25-mm 30-m fused silica capillary column coated with a 0.25-m film of G16 Temperature: See the table below.
Time (min) 0 55 75 Temperature 170 225 225
Injector: 250 Detector: 280 Carrier gas: Helium Split flow ratio: 1:200 Injection size: 1 L System suitability Samples: System suitability solution, Standard solution, and Sample solution Suitability requirements Resolution: NLT 1.3 between the peaks in the Standard solution due to methyl oleate and to methyl cis-vaccinate, and that between methyl gadoleate and methyl gondoate is sufficient for purposes of identification and area measurement Theoretical area percentages: 24.4 for methyl palmitate, 24.8 for methyl stearate, 25.2 for methyl arachidate, and 25.6 methyl behenate, System suitability solution Area percentages: Within 1% of the theoretical values from the System suitability solution System suitability criteria: The number of fatty acid methyl ester peaks exceeding 0.05% of the total area is at least 24, and the 24 largest peaks of the methyl esters account for more than 90% of the total area. (These correspond to the following, in common elution order:
USP 32
allow to stand until both layers are clear. Discard the lower layer. Wash the ether-hexane extracts by shaking vigorously with 50 mL of Solution C, and then washing with three 50mL portions of 10 mg/mL sodium chloride solution. Filter the upper layer through 5 g of anhydrous sodium sulfate on a fast filter paper into a 250-mL flask suitable for a rotary evaporator. Wash the filter with 10 mL of a mixture of ether and hexane (1:1), and combine with the extract. Evaporate the solvent at reduced pressure at a temperature not exceeding 30, and fill with nitrogen when the evaporation is complete. Alternatively evaporate the solvent under a gentle stream of nitrogen at a temperature not exceeding 30. Dissolve the residue in 1.5 mL of Mobile phase A. [NOTEGentle heating in an ultrasonic bath may be required. A large fraction of the white residue is cholesterol.] Sample solution B: To 4.00 g of Cod Liver Oil, add 2.0 mL of Internal standard solution, and proceed as directed for Sample solution A beginning with Add 5 mL of . Clean-up chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Mobile phase: Mobile phase A Column: 25-mm 4.6-cm; packing L10, stainless steel clean-up column Injection size: 350 L Analytical chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 265 nm Mobile phase: Mobile phase B Column: 15-mm 4.6-cm; 5 m packing L1, stainless steel Injection size: 200 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.4 between cholecalciferol and ergocalciferol Relative standard deviation: NMT 2.0% for the cholecalciferol peak response from five injections Analysis (Clean-up chromatographic system) Samples: Standard solution, Sample solution A, and Sample solution B Collect separately the eluates from 2 min before until 2 min after the retention time of cholecalciferol in a glass tube, containing 1 mL of Solution A and fitted with a hermetic closure. Evaporate each tube under a stream of nitrogen at a temperature not exceeding 30. Dissolve each residue in 1.5 mL of acetonitrile. Analysis (Analytical chromatographic system) Samples: Resulting solutions of Standard solution, Sample solution A, and Sample solution B Measure the peak responses at the retention times corresponding to cholecalciferol and ergocalciferol. Calculate the content of vitamin D, in g, in the portion of Cod Liver Oil taken: Result = 2 C (RU/RS) C RS RU = concentration of USP Cholecalciferol RS in the Standard solution (g/mL) = response of the cholecalciferol relative to the internal standard in the Standard solution = corrected relative response of Sample solution B calculated as follows: Result = rU2/[rIS2 (rIS1 rU2/rU1)] rU2
Official Monographs / Cod 171

= peak response for cholecalciferol in Sample solution A = peak response for cholecalciferol in Sample rU1 solution B = peak response for ergocalciferol in Sample rIS1 solution A = peak response for ergocalciferol in Sample rIS2 solution B Acceptance criteria: 1.5 g (60 USP Units) 6.25 g (250 USP Units) of vitamin D/g SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.9180.927 COLOR: When viewed transversely in a tall, cylindrical, standard oil-specimen bottle of colorless glass of about 120mL capacity, the color is not more intense than that of a mixture of cobaltous chloride CS, ferric chloride CS, and water (11:76:33), in a similar bottle of the same internal diameter. NONDESTEARINATED COD LIVER OIL Sample: Fill a tall, cylindrical, standard oil-specimen bottle of 120-mL capacity with Oil at a temperature between 23 and 28, insert the stopper, and immerse the bottle in a mixture of ice and water for 3 h. Acceptance criteria: The oil remains clear and does not deposit stearin. FATS AND FIXED OILS, Unsaponifiable Matter 401: NMT 1.30% FATS AND FIXED OILS, Acid Value 401 Sample: Mix 15 mL of alcohol with 15 mL of ether, add 5 drops of phenolphthalein TS, and neutralize with 0.1 N sodium hydroxide. Dissolve 2.0 g of Oil in the mixture, and boil the oil solution gently under a reflux condenser for 10 min. Analysis: Cool, and titrate the mixture with 0.1 N sodium hydroxide VS to the production of a pink color that persists after shaking for 30 s. Acceptance criteria: NMT 1.0 mL of 0.10 N sodium hydroxide is required. FATS AND FIXED OILS, Iodine Value 401: 145180 FATS AND FIXED OILS, Saponification Value 401: 180192 [NOTEIf carbon dioxide has been used as a preservative, expose the Oil in a shallow dish in a vacuum desiccator for 24 h before weighing the specimen for determination of the saponification value.] FATS AND FIXED OILS, Anisidine Value 401: NMT 30 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. It may be bottled or otherwise packaged in containers from which air has been expelled by the production of a vacuum or by an inert gas. LABELING: The vitamin A potency and vitamin D potency, when designated on the label, are expressed in USP Units/g of oil. The potencies may be expressed also in metric units, on the basis that 1 USP Vitamin A Unit equals 0.3 g and 40 USP Vitamin D Units equals 1 g. Where the content of docosahexaenoic acid or eicosapentaenoic acid are claimed, state their concentrations in mg/g. USP REFERENCE STANDARDS 11 USP Cholecalciferol RS USP Cod Liver Oil RS USP Ergocalciferol RS
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USP 32
Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, and allow the solvent to evaporate. Spray the plate with Spray reagent, and examine the chromatogram. Acceptance criteria: No spot obtained from Sample solution A, other than the principal spot and any spot observed at the origin, is more intense than the principal spot obtained from Sample solution B (2%); and NMT one such spot having an RF greater than that of the principal spot is more intense than the principal spot obtained from Sample solution C (1%). LIMIT OF MORPHINE Analysis: Dissolve 50 mg of potassium ferricyanide in 10 mL of water, add 1 drop of ferric chloride TS, and 1 mL of a neutral or slightly acid 10 mg/mL solution of Codeine prepared with the aid of sulfuric acid. Acceptance criteria: No blue color is produced immediately. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 154158 when previously dried The range between beginning and end of melting does not exceed 2. LOSS ON DRYING 731: Dry it at 80 for 4 h: it loses NMT 6.0% of its weight. READILY CARBONIZABLE SUBSTANCES 271 Sample Solution: Dissolve 10 mg in 5 mL of sulfuric acid TS. Acceptance criteria: The solution has no more color than Matching Fluid S. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Codeine Sulfate RS
Codeine
(Comment on this Monograph)id=m19560=Codeine=Chl-CyMonos.pdf)
C18H21NO3 H2O
317.38
C18H21NO3 299.37 Morphinan-6-ol, 7,8-didehydro-4,5-epoxy-3-methoxy-17methyl-, monohydrate, (5,6)-. 7,8-Didehydro-4,5-epoxy-3-methoxy-17-methylmorphinan-6ol monohydrate [6059-47-8]. Anhydrous [76-57-3]. DEFINITION Codeine, dried at 80 for 4 h, contains NLT 98.5% and NMT 100.5% of C18H21NO3. IDENTIFICATION A. INFRARED ABSORPTION 197K Standard: Dissolve 50 mg of USP Codeine Sulfate RS in 15 mL of water. Render it alkaline with 6 N ammonium hydroxide and extract with several 10-mL portions of chloroform. Evaporate the combined chloroform extracts to dryness on a steam bath, and dry the residue at 80 for 4 h. B. ULTRAVIOLET ABSORPTION 197U Solution: 100 g/mL in 0.1 N sulfuric acid Acceptance criteria: Absorptivity at 284 nm, calculated on the dried basis, is 112.9% to 119.9% of that of USP Codeine Sulfate RS. ASSAY PROCEDURE Sample solution: Dissolve 400 mg of Codeine, previously dried, by warming it in 30.0 mL of 0.1 N sulfuric acid VS. Cool. Add 10 mL of water, and add methyl red TS. Analysis: Titrate the excess acid in the Sample solution with 0.1 N sodium hydroxide VS. Perform a blank determination (see Titrimetry 541, Residual Titrations). Each mL of 0.1 N sulfuric acid is equivalent to 29.94 mg of C18H21NO3. Acceptance criteria: 98.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities CHROMATOGRAPHIC PURITY Sample solution A: 40 mg/mL of Codeine in dehydrated alcohol Sample solution B: 0.8 mg/mL of Codeine in dehydrated alcohol, from Sample solution A Sample solution C: 0.4 mg/mL of Codeine in dehydrated alcohol, from Sample solution A Developing solvent system: Dehydrated alcohol, cyclohexane, and ammonium hydroxide (72:30:6) Spray reagent: Mix 3 mL of chloroplatinic acid solution (1 in 10) with 97 mL of water. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 10 L Analysis Samples: Sample solution A, Sample solution B, and Sample solution C Apply each of the sample solutions to the plate. Allow the spots to dry, and develop the chromatogram in the
Codeine Phosphate
(Comment on this Monograph)id=m19590=Codeine Phosphate=Chl-Cy-Monos.pdf) C18H21NO3 H3PO4 1/2H2O 406.37 C18H21NO3 H3PO4 397.37 Morphinan-6-ol, 7,8-didehydro-4,5-epoxy-3-methoxy-17methyl-, (5,6)-, phosphate (1:1) (salt), hemihydrate; 7,8-Didehydro-4,5-epoxy-3-methoxy-17-methylmorphinan-6ol phosphate (1:1) (salt) hemihydrate [41444-62-6]. Anhydrous [52-28-8]. DEFINITION Codeine Phosphate contains NLT 99.0% and NMT 101.5% of C18H21NO3 H3PO4, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Analysis: Neutralize a 20 mg/mL solution with 6 N ammonium hydroxide, and add silver nitrate TS. Acceptance criteria: A yellow precipitate of silver phosphate is formed, and it is soluble in 2 N nitric acid and in 6 N ammonium hydroxide. ASSAY PROCEDURE Sample: 1 g of Codeine Phosphate Analysis: Dissolve the Sample in 50 mL of glacial acetic acid, warming slightly if necessary to effect solution. Titrate with 0.1 N perchloric acid VS. Perform a blank
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determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 39.74 mg of C18H21NO3 H3PO4. Acceptance criteria: 99.0%101.5% IMPURITIES Organic Impurities CHROMATOGRAPHIC PURITY Diluent: 0.01 N hydrochloric acid and dehydrated alcohol (4:1) Sample solution A: 40 mg/mL in Diluent Sample solution B: 0.8 mg/mL from Sample solution A in Diluent Sample solution C: 0.4 mg/mL from Sample solution A in Diluent Spray reagent: To a 3 mg/mL chloroplatinic acid solution and water add 60 mg/mL of potassium iodide solution (1:1) (See Chromatography 621, Thin-Layer Chromatography) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 10 L Developing solvent system: Dehydrated alcohol, cyclohexane, and ammonium hydroxide (12:5:1) Analysis Samples: Sample solution A, Sample solution B, and Sample solution C Apply each of the Sample solutions to the plate. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, and allow the solvent to evaporate. Spray the plate with Spray reagent, and examine the chromatogram. Acceptance criteria: No spot obtained from Sample solution A, other than the principal spot and any spot observed at the origin, is more intense than the principal spot obtained from Sample solution B (2%); and NMT one such spot having an RF greater than that of the principal spot is more intense than the principal spot from Sample solution C (1%). LIMIT OF MORPHINE Analysis: Dissolve 50 mg of potassium ferricyanide in 10 mL of water, add 1 drop of ferric chloride TS, and 1 mL of 10 mg/mL Codeine Phosphate solution. Acceptance criteria: No blue color is produced immediately. SPECIFIC TESTS ACIDITY Analysis: Dissolve 100 mg in 20 mL of water, and titrate with 0.010 N sodium hydroxide to a pH of 5.4, using a pH meter. Acceptance criteria: NMT 1.0 mL of 0.010 N sodium hydroxide is required. WATER DETERMINATION, Method I 921: NMT 3.0% CHLORIDE AND SULFATE, Chloride 221 Analysis: To 10 mL of 10 mg/mL of solution, acidified with nitric acid, add a few drops of silver nitrate TS. Acceptance criteria: No opalescence is produced immediately. CHLORIDE AND SULFATE, Sulfate 221 Analysis: To 10 mL of 10 mg/mL of solution, add a few drops of barium chloride TS. Acceptance criteria: No turbidity is produced immediately. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store up to 40 as permitted by the manufacturer. USP REFERENCE STANDARDS 11 USP Codeine Phosphate RS
Official Monographs / Codeine 173
Codeine Phosphate Injection

(Comment on this Monograph)id=m19600=Codeine Phosphate Injection=Chl-Cy-Monos.pdf) DEFINITION Codeine Phosphate Injection is a sterile solution of Codeine Phosphate in Water for Injection. It contains NLT 93.0% and NMT 107.0% of the labeled amount of C18H21NO3 H3PO4 1/2H2O. [NOTEDo not use the Injection if it is more than slightly discolored or contains a precipitate.] IDENTIFICATION A. INFRARED ABSORPTION Sample: Dilute a volume of Injection, equivalent to 90 mg of codeine phosphate, with water to 10 mL. Add 1 drop of hydrochloric acid, and extract with three 10-mL portions of chloroform, discarding the chloroform extracts. Add 6 N ammonium hydroxide until the solution is alkaline, and extract with several 10-mL portions of chloroform. Evaporate the combined chloroform extracts on a steam bath to dryness, and dry at 80 for 4 h. Acceptance criteria: The IR absorption spectrum of a potassium bromide dispersion of the Sample exhibits maxima at the same wavelengths as that of the codeine obtained by similarly treating 1 mL of 10 mg/mL solution of USP Codeine Phosphate RS. B. PROCEDURE Analysis: Neutralize the equivalent to 60 mg of codeine phosphate from Injection with 6 N ammonium hydroxide, and add silver nitrate TS. Acceptance criteria: A yellow precipitate of silver phosphate is formed, and is soluble in 2 N nitric acid and in 6 N ammonium hydroxide. ASSAY PROCEDURE Sample solution: Transfer the equivalent to 75 mg of codeine phosphate from Injection, to a small separator, and add 15 mL of water. Add 2 drops of phosphoric acid, and extract with four 10-mL portions of chloroform, collecting the chloroform extracts in a separator. Wash the combined chloroform extracts with 10 mL of water, and add the water wash to the first separator containing the sample. Discard the chloroform extracts, render the solution alkaline with 6 N ammonium hydroxide, and completely extract the alkaloid with successive 15-mL portions of chloroform. Evaporate the combined chloroform solution on a steam bath nearly to dryness. Dissolve the residue in 2 mL of methanol, heating if necessary, and add methyl red TS. Analysis: Titrate the Sample solution with 0.02 N sulfuric acid VS to a faint pink color. Add 40 mL of freshly boiled, cooled water, and complete the titration with 0.02 N sulfuric acid VS. Each mL of 0.02 N sulfuric acid is equivalent to 8.128 mg of C18H21NO3 H3PO4 1/2H2O. Acceptance criteria: 93.0%107.0%. IMPURITIES Organic Impurities LIMIT OF MORPHINE Analysis: Dissolve 50 mg of potassium ferricyanide in 10 mL of water, add 1 drop of ferric chloride TS, and 1 mL of a solution of 5 mg/mL of codeine phosphate. Acceptance criteria: No blue color is produced immediately. SPECIFIC TESTS PH 791: 3.06.0 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 5.8 USP Endotoxin Units/mg of codeine phosphate. OTHER REQUIREMENTS: It meets the requirements under Injections 1.
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Codeine Phosphate RS USP Endotoxin RS
Codeine Phosphate Tablets

(Comment on this Monograph)id=m19610=Codeine Phosphate Tablets=Chl-Cy-Monos.pdf) DEFINITION Codeine Phosphate Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of C18H21NO3 H3PO4 1/2H2O. IDENTIFICATION A. INFRARED ABSORPTION Sample: Digest the equivalent to 100 mg of codeine phosphate from finely powdered Tablets, with 15 mL of water and 5 mL of 2 N sulfuric acid for 1 h. Filter, if necessary, and wash any undissolved residue with a few mL of water. Render the filtrate alkaline with 6 N ammonium hydroxide, extract with several small portions of chloroform, evaporate the combined chloroform extracts on a steam bath to dryness, and dry at 80 for 4 h. Acceptance criteria: The IR absorption spectrum of a potassium bromide dispersion of the Sample exhibits maxima at the same wavelengths as that of the codeine obtained by similarly treating 1 mL of 10 mg/mL solution of USP Codeine Phosphate RS. B. PROCEDURE Analysis: To an equivalent of 100 mg of codeine phosphate from finely powdered Tablets, in 10 mL of water, add 2 drops of 2 N sulfuric acid. Digest, with frequent shaking, for 15 min. Neutralize 5 mL of the filtrate with 6 N ammonium hydroxide, and add silver nitrate TS. Acceptance criteria: A yellow precipitate of silver phosphate is formed, and it is soluble in diluted nitric acid and in 6 N ammonium hydroxide. [NOTESave a portion of the filtrate for the Limit of Morphine test.] ASSAY PROCEDURE Sample solution: Transfer an equivalent to 150 mg of codeine phosphate from finely powdered tablets (NLT 20 Tablets) to a 100-mL volumetric flask. Add 20 mL of 0.5 N sulfuric acid, and shake the mixture occasionally during 2 h. Add water to volume, mix, and filter through a filtering crucible. Transfer to a separator measured portion of the filtrate, equivalent to NLT 75 mg of codeine phosphate, render the solution alkaline with 6 N ammonium hydroxide, and completely extract the alkaloid with successive 15-mL portions of chloroform. Evaporate the combined chloroform solution on a steam bath nearly to dryness. Dissolve the residue in 2 mL of methanol, heating if necessary, and add methyl red TS. Analysis: Titrate with 0.02 N sulfuric acid VS to a faint pink color. Add 40 mL of freshly boiled, cooledwater, and complete the titration with 0.02 N sulfuric acid VS. Each mL of 0.02 N sulfuric acid is equivalent to 8.128 mg of C18H21NO3 H3PO4 1/2H2O.
IMPURITIES Organic Impurities LIMIT OF MORPHINE Analysis: Dissolve 50 mg of potassium ferricyanide in 10 mL of water, add 1 drop of ferric chloride TS and 1 mL portion of the filtrate from Identification test B. Acceptance criteria: No blue color is produced immediately. PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Detector: UV 284 nm Sample solutions: Sample per Dissolution 711 Filter and dilute with Medium to a concentration that is similar to that of the Standard solution Standard solution: USP Codeine Phosphate RS in Medium Tolerances: NLT 75% (Q) of the labeled amount of C18H21NO3 H3PO4 1/2H2O is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Content Uniformity Sample solution: Transfer 1 Tablet, previously crushed or finely powdered, to a 50-mL volumetric flask. Add 25 mL of water, and shake to dissolve. Dilute with water to volume, and filter, if necessary, discarding the first 20 mL of the filtrate. Transfer an aliquot of the filtrate, equivalent to 6 mg of codeine phosphate, to a 50-mL volumetric flask containing 2 mL of 3 N hydrochloric acid, and dilute with water to a volume-nominal concentration 0.12 mg/mL. Standard solution: 0.12 mg/mL of USP Codeine Phosphate RS in 0.1 N hydrochloric acid Spectrometric conditions Mode: UV-Vis Analytical wavelength: 284 nm Cell: 1 cm Blank: Water Analysis: Samples: Blank, Standard solution, and Sample solution Calculate the percentage of C18H21NO3 H3PO4 1/2H2O the Tablet taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 AU AS CS CU Mr1 Mr2 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Codeine Phosphate RS in the Standard solution (mg/mL) = nominal concentration of codeine phosphate in the Sample solution (mg/mL) = molecular weight of codeine phosphate hemihydrate, 406.37 = molecular weight of codeine phosphate, 397.37
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Codeine Phosphate RS
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Official Monographs / Codeine 175

spot is more intense than the principal spot from Sample solution C (1%). LIMIT OF MORPHINE Analysis: Dissolve 50 mg of potassium ferricyanide in 10 mL of water, and add 1 drop of ferric chloride TS and 1 mL of 10 mg/mL solution of Codeine Sulfate. Acceptance criteria: No blue color is produced immediately. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 112.5 to 115.0 Sample solution: 20 mg/mL, in water ACIDITY Analysis: Dissolve 500 mg in 15 mL of water, add 1 drop of methyl red TS, and titrate with 0.020 N sodium hydroxide. Acceptance criteria: NMT 0.30 mL is required for neutralization. WATER DETERMINATION, Method III 921: Dry 500 mg at 105 for 3 h: it loses between 6.0% and 7.5% of its weight. READILY CARBONIZABLE SUBSTANCES TEST 271 Sample solution: Dissolve 10 mg in 5 mL of sulfuric acid TS. Acceptance criteria: The solution has no more color than Matching Fluid S. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Codeine Sulfate RS
Codeine Sulfate
(Comment on this Monograph)id=m19640=Codeine Sulfate=Chl-Cy-Monos.pdf) (C18H21NO3)2 H2SO4 3H2O 750.85 696.82 (C18H21NO3)2 H2SO4 Morphinan-6-ol, 7,8-didehydro-4,5-epoxy-3-methoxy-17methyl-, (5,6)-, sulfate (2:1) (salt), trihydrate; 7,8-Didehydro-4,5-epoxy-3-methoxy-17-methylmorphinan-6ol sulfate (2:1) (salt) trihydrate [6854-40-6]. Anhydrous [1420-53-7]. DEFINITION Codeine Sulfate, dried at 105 for 3 h, contains NLT 98.5% and NMT 100.5% of (C18H21NO3)2 H2SO4. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Solution: 100 g/mL in water Acceptance criteria: Absorptivities at 284 nm, do not differ by more than 3.0%, calculated on the dried basis. C. CHLORIDE AND SULFATE, Sulfate 191 ASSAY PROCEDURE Sample: 1.4 g of Codeine Sulfate Analysis: Dissolve the Sample, previously dried, in 50 mL of glacial acetic acid, warming if necessary, to effect solution. Titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any necessary correction (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 69.68 mg of (C18H21NO3)2 H2SO4. Acceptance criteria: 98.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities CHROMATOGRAPHIC PURITY Diluent: 0.01 N hydrochloric acid and dehydrated alcohol (4:1) Sample solution A: 40 mg/mL in Diluent Sample solution B: 0.8 mg/mL from Sample solution A in Diluent Sample solution C: 0.4 mg/mL from Sample solution A in Diluent Spray reagent: 3 mg/mL chloroplatinic acid solution and 60 mg/mL potassium iodide solution (1:1) Chromatographic System (See Chromatography 621, ThinLayer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 10 L Developing solvent system: Dehydrated alcohol, cyclohexane, and ammonium hydroxide (12:5:1) Analysis Samples: Sample solution A, Sample solution B, and Sample solution C Apply each of the Sample solutions to the plate. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent front has moved three-fourths of the length of the plate. Remove the plate from the developing chamber, and allow the solvent to evaporate. Spray the plate with Spray reagent, and examine the chromatogram. Acceptance criteria: No spot obtained from Sample solution A, other than the principal spot and any spot observed at the origin, is more intense than the principal spot obtained from Sample solution B (2%); and NMT one such spot having an RF greater than that of the principal
Codeine Sulfate Tablets

(Comment on this Monograph)id=m19670=Codeine Sulfate Tablets=Chl-Cy-Monos.pdf) DEFINITION Codeine Sulfate Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of (C18H21NO3)2 H2SO4 3H2O. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Digest an equivalent to 50 mg of codeine sulfate from finely powdered Tablets, with 15 mL of water and 5 mL of 2 N sulfuric acid for 1 h. Filter, if necessary, and wash any undissolved residue with a few mL of water. Render the filtrate alkaline with 6 N ammonium hydroxide, extract with several small portions of chloroform, and evaporate the chloroform solution on a steam bath to dryness, drying at 80 for 4 h. Standard: 50 mg of USP Codeine Sulfate RS dissolved in 15 mL of water, then rendered alkaline with 6 N ammonium hydroxide and extracted with several 10-mL portions of chloroform, followed by evaporation of the combined chloroform extracts on a steam bath to dryness, drying at 80 for 4 h. B. IDENTIFICATION TESTS-GENERAL, Sulfate 191: A filtered solution of Tablets meets the requirements. ASSAY PROCEDURE Sample solution: Transfer an equivalent to 150 mg of codeine sulfate from finely powdered Tablets (NLT 20 Tablets) to a 100-mL volumetric flask. Add sufficient water to make a thin suspension, then add 20 mL of 0.5 N sulfuric acid. Shake the mixture occasionally for 2 h, and allow to stand for 16 h. Dilute with water to volume, mix, and filter through a filtering crucible. Transfer measured portion of the
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filtrate, equivalent to NLT 75 mg of codeine sulfate, to a separator. Render the solution alkaline with 6 N ammonium hydroxide, and completely extract the alkaloid with successive 15-mL portions of chloroform. Evaporate the combined chloroform solution on a steam bath nearly to dryness, add 25.0 mL of 0.02 N sulfuric acid VS, and heat gently to dissolve the codeine and expel all of the chloroform. Cool and add methyl red TS. Analysis: Titrate the excess acid with 0.02 N sodium hydroxide VS. Each mL of 0.02 N sulfuric acid is equivalent to 7.509 mg of (C18H21NO3)2 H2SO4 3H2O. Acceptance criteria: 93.0%107.0% PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 500 mL Apparatus 1: 100 rpm Time: 45 min Detector: UV 284 nm Sample solutions: Sample per Dissolution 711 Filter and dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: USP Codeine Sulfate RS in Medium Tolerances: NLT 75% (Q) of the labeled amount of (C18H21NO3)2 H2SO4 3H2O is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Content uniformity Sample solution: Transfer 1 Tablet to a 50-mL volumetric flask. Add 20 mL of 0.5 N sulfuric acid and 10 mL of water. Shake until the Tablet is disintegrated, and allow to stand for 16 h. Dilute with water to volume, and filter, discarding the first few mL of the filtrate. Dilute the resulting filtrate with 0.2 N sulfuric acid to obtain a solution containing nominally 120 g/mL of codeine sulfate (trihydrate). Standard solution: 110 g/mL of USP Codeine Sulfate RS in 0.2 N sulfuric acid Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 284 nm Cell: 1 cm Blank: 0.2 N sulfuric acid Analysis Samples: Blank, Standard solution, and Sample solution Calculate the percentage of (C18H21NO3)2 H2SO4 3H2O in the Tablet taken: Result = (AU/AS) (CS/CU) (Mr1/Mr2) 100 AU AS CS CU Mr1 Mr2 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of the USP Codeine Sulfate RS in the Standard solution (g/mL) = nominal concentration of codeine sulfate trihydrate in the Sample solution (g/mL) = molecular weight of codeine sulfate trihydrate, 750.87 = molecular weight of anhydrous codeine sulfate, 696.82
Colchicine
(Comment on this Monograph)id=m19720=Colchicine=Chl-CyMonos.pdf)
399.44 C22H25NO6 Acetamide, N-(5,6,7,9-tetrahydro-1,2,3,10-tetramethoxy-9oxobenzo[a]heptalen-7-yl)-, (S)-; Colchicine [64-86-8]. DEFINITION Colchicine is an alkaloid contained in various species of Colchicum and in other genera. It contains NLT 94.0% and NMT 101.0% of C22H25NO6, calculated on the anhydrous, solvent-free basis. [CAUTIONColchicine is extremely poisonous.] IDENTIFICATION INFRARED ABSORPTION 197K [NOTEDisregard any peak occurring at 1735 cm1.] ASSAY PROCEDURE [NOTEPerform all dilutions in low-actinic glassware.] Mobile phase: Dilute 45 mL of 0.5 M monobasic potassium phosphate with water to 450 mL. Add 530 mL of methanol, cool to room temperature, and add methanol to bring the volume to 1000 mL. Adjust with 0.5 M phosphoric acid to a pH of 5.5 0.05, and pass through a 0.45-m membrane filter. Standard solution: 6 g/mL of USP Colchicine RS in a mixture of methanol and water (1:1) [NOTEThis solution is stable for 4 months when stored tightly stoppered and in the dark.] Sample solution: 6 g/mL of Colchicine in a mixture of methanol and water (1:1) [NOTEPrepare immediately before use.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254-nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1.0 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 4500 theoretical plates Retention time: 5.59.5 min for colchicine Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Measure the responses for all peaks recorded during 1.5 times the retention time for colchicine.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Codeine Sulfate RS
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Calculate the percentage of C22H25NO6 in the Colchicine taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Colchicine RS in the Standard solution (g/mL) = nominal concentration of cholchicine in the CU Sample solution (g/mL) Acceptance criteria: 94.0%101.0% IMPURITIES Organic Impurities LIMIT OF COLCHICEINE Analysis: To 5 mL of 10 mg/mL solution, add 2 drops of ferric chloride TS. Acceptance criteria: No definite green color is produced. LIMIT OF ETHYL ACETATE Internal standard solution: Dilute 0.5 mL of n-propyl alcohol with water to 100.0 mL. Standard solution: Pipet 1 mL of ethyl acetate and 0.5 mL of n-propyl alcohol into a 1000-mL volumetric flask, and add water to volume. This solution contains 0.90 mg/mL of ethyl acetate. Sample solution: Place 250 mg of Colchicine in a 10-mL volumetric flask, dissolve in 8 mL of water, and add 1.0 mL of Internal standard solution. Add water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 1.5-m column packed with 20% (w/v) phase G14 on support S1 Column temperature: 75 Carrier gas: Nitrogen Analysis Samples: Standard solution and Sample solution Determine the peak height for ethyl acetate relative to the peak height for n-propyl alcohol, and calculate the percentage, by weight, of ethyl acetate in the portion of Colchicine taken. Acceptance criteria: NMT 8.0% CHROMATOGRAPHIC PURITY Procedure: Proceed as directed in the Assay. Acceptance criteria Sum of impurities: NMT 5.0% other than that due to colchicine, eluting within 1.5 times the retention time for colchicine SPECIFIC TESTS SPECIFIC ROTATION 781S: 240 to 250, calculated on the anhydrous, solvent-free basis Sample solution: 10 mg/mL, in alcohol WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Colchicine RS rU rS CS
Official Monographs / Colchicine 177

Hydroxide. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H25NO6. [CAUTIONColchicine is extremely poisonous.] IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Transfer the equivalent to 2 mg of colchicine from a volume of Injection, to a separator. Add 5 mL of water, and extract with 15 mL of chloroform. Evaporate the chloroform extract to dryness using mild heat. Acceptance criteria: The IR absorption spectrum of a potassium bromide dispersion of the Sample exhibits maxima only at the same wavelengths as that of a similar preparation of USP Colchicine RS. B. ULTRAVIOLET ABSORPTION 197U Sample solution: 10 g/mL of colchicine from Injection, in water Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Colchicine RS, concomitantly measured. ASSAY PROCEDURE [NOTEPerform all dilutions in low-actinic glassware.] Mobile phase: Dilute 45 mL of 0.5 M monobasic potassium phosphate with water to 450 mL. Add 530 mL of methanol, cool to room temperature, and add methanol to bring the volume to 1000 mL. Adjust with 0.5 M phosphoric acid to a pH of 5.5 0.05, and pass through a 0.45-m membrane filter. Standard solution: 6 g/mL of USP Colchicine RS in a mixture of methanol and water (1:1) [NOTEThis solution is stable for 4 months when stored tightly stoppered and in the dark.] Sample solution: 6 g/mL of Colchicine from a measured volume of Injection, in a mixture of methanol and water (1:1) [NOTEPrepare immediately before use.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254-nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1.0 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 4500 theoretical plates Retention time: 5.59.5 mins for colchicine Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Measure the responses for all peaks recorded during 1.5 times the retention time for colchicine. Calculate the percentage of C22H25NO6 in the Injection taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Colchicine RS in the Standard solution (g/mL) = nominal concentration of colchicine in the Sample solution (g/mL)
Colchicine Injection
(Comment on this Monograph)id=m19730=Colchicine Injection=Chl-Cy-Monos.pdf) DEFINITION Colchicine Injection is a sterile solution of C22H25NO6 in Water for Injection, prepared from Colchicine with the aid of Sodium
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90.0%110.0%
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Mode: LC Detector: UV 254-nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1.0 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 4500 theoretical plates Retention time: 5.59.5 min for colchicine Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Measure the responses for all peaks recorded during 1.5 times the retention time for colchicine. Calculate the percentage of C22H25NO6 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of the USP Colchicine Rs in the Standard solution (g/mL) = nominal concentration of colchicine in the CU Sample solution (g/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711: [NOTEConduct this procedure without delay, under subdued light, and using low-actinic glassware.] Medium: Water; 500 mL Apparatus 1 : 100 rpm Time: 30 min Analysis: Determine the amounts of C22H25NO6 dissolved, by employing the procedure in the Assay, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amount of C22H25NO6. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Colchicine RS
SPECIFIC TESTS PH 791: 6.07.2, in a solution of Injection containing 1.0 mg/mL of potassium chloride BACTERIAL ENDOTOXINS TEST 85: Contains NMT 166.7 USP Endotoxin Units mg of colchicine OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Colchicine RS USP Endotoxin RS
Colchicine Tablets
(Comment on this Monograph)id=m19740=Colchicine Tablets=Chl-Cy-Monos.pdf) DEFINITION Colchicine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C22H25NO6. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Triturate the equivalent of 20 mg of colchicine, from ground tablets, with 20 mL of water, allow the solids to settle, and filter the supernatant into a separator. Extract with 30 mL of chloroform. Evaporate the chloroform extract to dryness using mild heat. Acceptance criteria: The IR absorption spectrum of a potassium bromide dispersion of the Sample exhibits maxima only at the same wavelengths as that of a similar preparation of USP Colchicine RS. ASSAY PROCEDURE [NOTEPerform all dilutions in low-actinic glassware.] Mobile phase: Dilute 45 mL of 0.5 M monobasic potassium phosphate with water to 450 mL. Add 530 mL of methanol, cool to room temperature, and add methanol to bring the volume to 1000 mL. Adjust with 0.5 M phosphoric acid to a pH of 5.5 0.05, and pass through a 0.45-m membrane filter. Standard solution: 6 g/mL of USP Colchicine RS in a mixture of methanol and water (1:1) [NOTEThis solution is stable for 4 months when stored tightly stoppered and in the dark.] Sample solution: Transfer the equivalent to 0.6 mg of colchicine from finely powdered Tablets (NLT 20 Tablets), to a 100-mL volumetric flask, add 50 mL of a mixture of methanol and water (1:1), and shake by mechanical means for 15 min, rinsing down the walls of the flask at about 8 min. Dilute with the same mixture to volume, and pass through a 0.45 m filter. [NOTEPrepare immediately before use.] Chromatographic system (See Chromatography 621, System Suitability.)
Colestipol Hydrochloride
(Comment on this Monograph)id=m19830=Colestipol Hydrochloride=Chl-Cy-Monos.pdf) DEFINITION Colestipol Hydrochloride is an insoluble, high molecular weight basic anion-exchange copolymer of diethylenetriamine and 1chloro-2,3-epoxypropane with approximately one out of five amino nitrogens protonated. Each gram binds NLT 1.1 mEq and NMT 1.6 mEq of sodium cholate, calculated as cholate binding capacity.
USP 32
IDENTIFICATION PYROLYSIS GAS CHROMATOGRAPHY Standard solution: Transfer an appropriate amount of USP Colestipol Hydrochloride RS into the probe. If necessary to keep the colestipol hydrochloride in the probe, mix four parts with one part of n-eicosane. Grind in a mortar with chloroform until the colestipol hydrochloride is uniformly coated with the n-eicosane. [NOTEThis solution is stable indefinitely but may require wetting with a small amount of chloroform before each use.] Sample solution: Proceed as directed under Standard solution, using an appropriate amount of the Sample specimen. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 3-mm 180-cm column packed with 80- to 100mesh support S1A which has been coated with 0.25% potassium hydroxide and 5% phase G16 Column temperature: 85 Detector temperature: 270 Carrier gas: Helium Flow rate: 60 mL/min [NOTEThe pyrolysis unit is capable of reaching 1100 when equipped with a platinum probe; pyrolysis time is NLT 10 s.] Analysis: Install the pyrolysis unit on the chromatograph, and position the probe so that it is just above but not touching the column packing. Set the pyrolysis temperature to about 1100. Separately pyrolyze the Standard solution and the Sample solution. After the completion of each pyrolysis cycle, remove and clean the probe. Acceptance criteria: The pyrogram of the Sample solution corresponds to that of the Standard solution obtained the same day. OTHER COMPONENTS CONTENT OF CHLORIDE Sample solution: Using 20 mg of Colestipol Hydrochloride proceed as directed under Oxygen Flask Combustion 471, and use 10 mL of 0.05 N sodium hydroxide as the absorbing liquid. Do not allow the paper specimen wrapper to come into contact with the liquid, and ignite the paper with an IR igniter. After combustion is complete, shake the flask vigorously, and allow to stand, with frequent shaking, for 40 min or until no cloudiness is present. Transfer the solution to a 50-mL beaker. Wash the flask with two 5-mL portions of water and two 10-mL portions of alcohol, adding each washing to the beaker, then add 0.2 mL of nitric acid. Reagent blank solution: Using a paper specimen wrapper, complete the combustion, and allow the mixture to stand for 40 min or until no cloudiness is present, as directed under Sample solution. Transfer the solution so obtained to a 50-mL beaker. Wash the combustion flask with two 5-mL portions of water and two 10-mL portions of alcohol, adding the washings to the beaker, then add 0.2 mL of nitric acid. Analysis: Titrate the Sample solution and the Reagent blank solution with 0.05 N silver nitrate VS, determining the endpoint potentiometrically, using a silver-silver chloride electrode and a glass reference electrode (see Titrimetry 541). Determine the volume of 0.05 N silver nitrate VS consumed by the Sample specimen taken: Result = V VB V = volume of titrant used for the Sample solution (mL) VB
Official Monographs / Colestipol 179

= volume of titrant used for the Reagent blank solution (mL) Each mL of 0.05 N silver nitrate is equivalent to 1.773 mg of Cl. Acceptance criteria: 6.5%9.0%, calculated on the dried basis
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.3% HEAVY METALS, Method II 231: NMT 20 ppm SPECIFIC TESTS PH 791: 6.07.5 Sample solution: Prepare a 10% (w/w) suspension of it in deionized water in a clean vial. Insert the stopper, shake at approximately 10-min intervals for 1 h, and centrifuge. Transfer a portion of the clear supernatant to a suitable container, and record the pH as soon as the reading has stabilized. LOSS ON DRYING 731: Dry it in vacuum at a pressure of 5 mm of mercury at 75 for 16 h: it loses NMT 1.0% of its weight. WATER ABSORPTION Sample solution: Transfer 5 g of Colestipol Hydrochloride to a dry, 100-mL plastic container, and add 80 g of water. Cover the container, and allow the suspension to equilibrate for 72 h. Analysis: With the aid of vacuum, filter the slurry transferred to a medium-porosity, fritted-glass funnel, and collect the filtrate in a tared plastic container. Disconnect the vacuum 2 min after the collection of the last portion of the filtrate. Weigh the container and the filtrate, and determine the weight of the filtrate in grams. Determine the amount of water absorbed by subtracting the weight of the filtrate from the weight of water taken for the test, and divide the weight, in grams, of the absorbed water by the weight, in grams, of Colestipol Hydrochloride taken. Acceptance criteria: Each gram absorbs 3.3 g5.3 g of water. CHOLATE BINDING CAPACITY Solution A: 10.0 mg/mL of sodium cholate and 9.0 mg/mL of sodium chloride, in water. Adjust the solution by the dropwise addition of hydrochloric acid to a pH of 6.4 0.1. Sample solution: Transfer 1.0 0.01 g of Colestipol Hydrochloride to a glass-stoppered, 125-mL conical flask. Add 100.0 mL of Solution A, insert the stopper securely in the flask, shake vigorously for 90 min with the flask positioned horizontally on a platform shaker, remove the flask from the shaker, and allow the solids to settle for 5 min. Analysis: Transfer 20.0 mL of supernatant from the Sample solution to a 40-mL beaker and transfer 20.0 mL of Solution A to a second 40-mL beaker. Adjust both solutions by the dropwise addition of 1 N sodium hydroxide to a pH of 10.5 0.5. Titrate both solutions with 0.1 N hydrochloric acid VS, determining the endpoints potentiometrically, and measure the titrant volume corresponding to the difference between the midpoints of the two inflections in the titration curves obtained for each solution (see Titrimetry 541). Determine the volume of titrant equivalent to the bound cholate by subtracting the volume of 0.1 N hydrochloric acid VS used in titrating the Sample solution from that used in titrating Solution A. Calculate the Cholate binding capacity, in mEq/g: Result = 5VN/W V N = volume of titrant equivalent to the bound cholate (mL) = normality of the 0.1 N hydrochloric acid VS
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USP 32
= weight of Colestipol Hydrochloride taken for the Sample solution (g) Acceptance criteria: 1.1 mEq/g 1.6 mEq/g. WATER-SOLUBLE SUBSTANCES Sample: 5.0 g of Colestipol Hydrochloride Analysis: Transfer the Sample to a glass-stoppered, 125-mL conical flask, add 80.0 mL of water, insert the stopper in the flask, and mount the flask in a water-bath shaker maintained at 37 1. Operate the shaker for 72 h, remove the flask from the shaker, and filter the contents through a fineporosity, fritted-glass funnel, collecting the filtrate in a tared 125-mL conical flask. Rinse any residual test material in the flask with two 5-mL portions of water, pass the washings through the filter, and combine the filtrates from the washings with the filtrate from the Sample mixture. Evaporate the filtrate to dryness, using filtered air or nitrogen if necessary, to aid in the evaporation. Dry the residue in a vacuum oven maintained at 75 for 1 h, allow to cool in a desiccator, and weigh. Acceptance criteria: NMT 0.5% COLESTIPOL EXCHANGE CAPACITY Resin base solution: Combine NLT 2 g of Colestipol Hydrochloride and 100 mL of 1 N sodium hydroxide in a 125-mL conical flask, insert a stopper in the flask, secure the flask on a platform shaker, and shake the mixture for 3 to 4 h. Filter the suspension through a coarse-porosity, frittedglass funnel, and wash the resin with 500 mL of water. Transfer the resin to a 1000-mL beaker, add 200 mL of water, allow to stand for 10 min, filter the suspension, and check the pH of the filtrate. Repeat the washing procedure with 200-mL portions of water until the pH of the filtrate is below 8 (as much as 5000 mL of water may be required). Dry the colestipol base resin so obtained and the funnel at a pressure of about 5 mm of mercury at 60 for 16 h. Break up any aggregates, and store the Resin base solution in a desiccator. Analysis: Transfer 1.0 g of the Resin base solution to a 125mL conical flask, add 100.0 mL of 0.20 N hydrochloric acid, insert a stopper in the flask, and shake the mixture by mechanical means for 2.5 h. Filter a portion of the suspension through a pledget of glass wool, and transfer 8.0 mL of the filtrate (Sample solution) to a 25-mL beaker. Transfer 5.0 mL of the same 0.20 N hydrochloric acid that was used to equilibrate the resin to a second 25-mL beaker, and add 5.0 mL of water. Titrate both solutions with 0.2 N sodium hydroxide VS, determining the endpoints potentiometrically (see Titrimetry 541), and calculate the Colestipol exchange capacity, taken in mEq/g: Result = (100N/W) [(Vb/5) (Va/8)] N W Vb = normality of the sodium hydroxide VS = weight of the Resin base solution taken (g) = volume of titrant used to neutralize the 5.0-mL aliquot of 0.20 N hydrochloric acid (mL) = volume of titrant used to neutralize the residual Va acid in the Sample solution (mL) Acceptance criteria: Each gram exchanges 9.0 11.0 mEq of sodium hydroxide, calculated as colestipol exchange capacity.
Colestipol Hydrochloride for Oral Suspension

(Comment on this Monograph)id=m19860=Colestipol Hydrochloride for Oral Suspension=Chl-Cy-Monos.pdf) DEFINITION Colestipol Hydrochloride for Oral Suspension is a mixture of Colestipol Hydrochloride with a suitable flow-promoting agent. Each g binds NLT 1.1 mEq and NMT 1.6 mEq of sodium cholate, calculated as the cholate binding capacity. IDENTIFICATION PYROLYSIS GAS CHROMATOGRAPHY Standard solution: Transfer an appropriate amount of USP Colestipol Hydrochloride RS into the probe. If necessary to keep the colestipol hydrochloride in the probe, mix four parts with one part n-eicosane. Grind in a mortar with chloroform until the colestipol hydrochloride is uniformly coated with the n-eicosane. [NOTEThis solution is stable indefinitely but may require wetting with a small amount of chloroform before each use.] Sample solution: Proceed as directed under Standard solution, using an appropriate amount of the Colestipol Hydrochloride for Oral Suspension. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 3-mm 180-cm column packed with 80- to 100mesh support S1A which has been coated with 0.25% potassium hydroxide and 5% phase G16 Temperature Column: 85 Detector: 270 Carrier gas: Helium Flow rate: 60 mL/min [NOTEThe pyrolysis unit is capable of reaching 1100 when equipped with a platinum probe, and pyrolysis time is NLT 10 s.] Analysis: Install the pyrolysis unit on the chromatograph, and position the probe so that it is just above but not touching the column packing. Set the pyrolysis temperature to about 1100. Separately pyrolyze the Standard solution and the Sample solution. After the completion of each pyrolysis cycle, remove and clean the probe. Acceptance criteria: The pyrogram of the Sample solution corresponds to that of the Standard solution obtained the same day. PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements for powders SPECIFIC TESTS WATER-SOLUBLE SUBSTANCES Sample: 5.0 g of Colestipol Hydrochloride for Oral Suspension Analysis: Transfer the Sample to a glass-stoppered, 125-mL conical flask, add 80.0 mL of water, insert the stopper in the flask, and mount the flask in a water-bath shaker maintained at 37 1. Operate the shaker for 72 h, remove the flask from the shaker, and filter the contents through a fineporosity, fritted-glass funnel, collecting the filtrate in a tared 100-mL fused quartz crucible. Rinse any residual sample
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Colestipol Hydrochloride RS
USP 32
material in the flask with two 5-mL portions of water, pass the washings through the filter, and combine the filtrates from the washings with the filtrate from the sample mixture. Evaporate the filtrate to dryness, filtered air or nitrogen being used, if necessary, to aid in the evaporation. Dry the residue in a vacuum oven maintained at 75 for 1 h, allow to cool in a desiccator, and weigh. Calculate the initial percentage of water-soluble substances in the portion of Colestipol Hydrochloride for Oral Suspension taken. Again heat the residue in a muffle furnace maintained at 800 25 for 4 h, allow to cool in a desiccator, and weigh. Calculate the percentage of inert ingredients present. Calculate the actual percentage of water-soluble substances in the portion of Colestipol Hydrochloride for Oral Suspension taken by subtracting the percentage of inert ingredients from the initial percentage of water-soluble substances found. Acceptance criteria: NMT 0.5% PH 791: 6.07.5 Sample solution: Prepare a 10% (w/w) suspension of Colestipol Hydrochloride for Oral Suspension in deionized water in a clean vial. Insert the stopper, shake at approximately 10-min intervals for 1 h, and centrifuge. Transfer a portion of the clear supernatant to a suitable container, and record the pH as soon as the reading has stabilized CHOLATE BINDING CAPACITY Solution A: 10.0 mg/mL of sodium cholate and 9.0 mg/mL of sodium chloride in water Adjust the solution by the dropwise addition of hydrochloric acid to a pH of 6.4 0.1. Sample solution: 1.0 0.01 g of Colestipol Hydrochloride for Oral Suspension to a glass-stoppered, 125-mL conical flask Add 100.0 mL of Solution A, insert the stopper securely in the flask, shake vigorously for 90 min with the flask positioned horizontally on a platform shaker, remove the flask from the shaker, and allow the solids to settle for 5 min. Analysis: Transfer 20.0 mL of supernatant from the Sample solution to a 40-mL beaker, transfer 20.0 mL of Solution A to a second 40-mL beaker, and adjust both solutions by the dropwise addition of 1 N sodium hydroxide to a pH of 10.5 0.5. Titrate both solutions with 0.1 N hydrochloric acid VS, determining the endpoints potentiometrically, and measure the titrant volume corresponding to the difference between the midpoints of the two inflections in the titration curves obtained for each solution (see Titrimetry 541). Determine the volume of titrant equivalent to the bound cholate by subtracting the volume of 0.1 N hydrochloric acid VS used in titrating the Sample solution from that used in titrating the Solution A Calculate the Cholate binding capacity, in mEq/g: Result = 5VN/W = volume of titrant equivalent to the bound cholate (mL) N = normality of the 0.1 N hydrochloric acid VS W = weight of Colestipol Hydrochloride taken for the Sample solution (g) Acceptance criteria: 1.1 1.6 mEq/g. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, single-dose or multiple-dose containers. USP REFERENCE STANDARDS 11 USP Colestipol Hydrochloride RS V
Official Monographs / Colestipol 181
Colestipol Hydrochloride Tablets

(Comment on this Monograph)id=m19865=Colestipol Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Colestipol Hydrochloride Tablets contain Colestipol Hydrochloride. Each Tablet binds NLT 1.1 mEq and NMT 1.6 mEq of sodium cholate/g of the labeled amount of colestipol hydrochloride, calculated as cholate binding capacity. IDENTIFICATION INFRARED ABSORPTION 197K Sample solution: Completely remove the coating film from a Tablet with a suitable implement, and grind the contents into a fine powder. To 3040 mg of the powder, add 15 mL of methanol. Shake vigorously for 3 min, then sonicate for 10 min, and shake for another 3 min. Pass through a suitable paper filter, wash the residue three times, each time with 10 mL of methanol. [NOTEA qualitative paper filter, with a coarse porosity and a particle retention of 20-25 m, is suitable.] Dry the residue at 60 in vacuum for 2 h. Mix 4 mg of the dried sample with 150 mg of potassium bromide. Standard solution: Mix 34 mg of USP Colestipol Hydrochloride RS with 150 mg of potassium bromide. PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 Content Uniformity Solution A, Solution B, Solution C, Mobile phase, Standard solution, and Chromatographic system: Proceed as directed in the test for Cholate Binding Capacity. Sample solution: Transfer 1 Tablet to a 100-mL volumetric flask, dilute with Solution C to volume, and stir for 120 min. Let the sample settle for at least 10 min, and filter a portion using a 0.45-m PVDF filter, discarding the first 5 mL of the filtrate. Analysis: Proceed as directed in the test for Cholate Binding Capacity. Select NLT 30 Tablets. Test 10 Tablets individually as directed above. Acceptance criteria: The requirements are met if the cholate binding capacity in each of the 10 Tablets lies within the range of 1.151.55 mEq/g of the labeled amount of colestipol hydrochloride, and the relative standard deviation is NMT 6.0%. If one Tablet is outside the range of 1.151.55 mEq/g and no Tablet is outside the range of 1.011.69 mEq/g, or if the relative standard deviation is greater than 6.0%, or if both conditions prevail, test 20 additional Tablets. The requirements are met if NMT one Tablet of the 30 is outside the range of 1.151.55 mEq/g and no Tablet is outside the range of 1.011.69 mEq/g of the labeled amount of colestipol hydrochloride, and the relative standard deviation for 30 Tablets does not exceed 7.8%. SPECIFIC TESTS PH 791: 5.57.5 Sample solution: Transfer 5 g of ground Tablets to a suitable flask, add 50 mL of deionized water, close the flask with a stopper, and stir for 30 min or until the Tablets completely disintegrate. Centrifuge to obtain a clear supernatant, and test pH on the clear supernatant. CHOLATE BINDING CAPACITY Solution A: 12.5 mg/mL of monobasic sodium phosphate. Adjust with phosphoric acid to a pH of 2.5 0.05. Mobile phase: Solution A, acetonitrile, and methanol (50:36:14)
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Solution B: 9.0 mg/mL of sodium chloride Solution C: Transfer a quantity of sodium cholate to a suitable volumetric flask, add Solution B to 80% of the final volume, sonicate to dissolve, and dilute with Solution B to volume to obtain a solution having a known concentration of 10.0 mg/mL of sodium cholate on the anhydrous basis. [NOTEDetermine the water content of sodium cholate titrimetrically at the time of use.][NOTEAdjust the solution by the dropwise addition of 0.5 N hydrochloric acid to a pH of 6.45 + 0.05.] [NOTEDo not allow the pH to go below 6.40 at any time.] Use this solution as soon as possible after preparation. Sample solution: Transfer 10 Tablets to a glass-stoppered 1.5-L flask. Add 1000.0 mL of Solution C, insert the stopper securely in the flask, and stir for 120 min. Filter a portion using a 0.45-m PVDF filter, discarding the first 5 mL of the filtrate. Standard solution: 4.0 mg/mL of sodium cholate from Solution C in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; 5-m packing L7 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the concentration of the unbound cholate, CT, in the Sample solution: CT = (RU/RS) CS RU RS CS = peak area from the Sample solution = peak area from the Standard solution = concentration of sodium cholate in the Standard solution (mg/mL) Calculate the cholate binding capacity, in mEq/g, of the labled amount of colestipol hydrochloride: Result = (CCH CT/NL) (F/Mr) CCH N L F Mr = concentration of sodium cholate in the Solution C (mg/mL) = number of Tablets taken to prepare the Sample solution = labeled amount of colestipol hydrochloride (g/Tablet) = conversion coefficient to g, 1000 = molecular weight of sodium cholate, 430.6
Colistimethate Sodium
(Comment on this Monograph)id=m19880=Colistimethate Sodium=Chl-Cy-Monos.pdf)
C58H105N16Na5O28S5 (colistin A component) C57H103N16Na5O28S5 (colistin B component) Colistimethate sodium; Pentasodium colistinmethanesulfonate [8068-28-8; 21362-08-3].
1749.82 1735.80
DEFINITION Colistimethate Sodium has a potency equivalent to NLT 390 g of colistin mg. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample solution: Dissolve a suitable quantity of Colistimethate Sodium in 2.0 mL of water, add a sufficient, measured volume of Buffer No. 6 to obtain a solution having a convenient concentration. Analysis: Proceed as directed for AntibioticsMicrobial Assays 81, Colistimethate Sodium, using a measured volume of Sample solution diluted with Buffer No. 6 to yield a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: NLT 390 g of colistin mg IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231:
NMT 0.003%
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Colestipol Hydrochloride RS
SPECIFIC TESTS PH 791: 6.58.5, in 10 mg/mL solution LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 7.0% of its weight. FREE COLISTIN Sample: Dissolve 80 mg in 3 mL of water, and add 0.05 mL of 100 mg/mL of silicotungstic acid solution. Acceptance criteria: No immediate precipitate is formed. CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. STERILITY TESTS 71: Meets the requirements when tested as directed for Membrane Filtration, Test for Sterility of the Product to be Examined BACTERIAL ENDOTOXINS TEST 85: Where the label states that Colistimethate Sodium is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 2.0 USP Endotoxin Units mg of colistin.
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Colistimethate Sodium RS USP Endotoxin RS
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ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. USP REFERENCE STANDARDS 11 USP Colistimethate Sodium RS USP Endotoxin RS
Colistin Sulfate
(Comment on this Monograph)id=m19920=Colistin Sulfate=Chl-Cy-Monos.pdf)
Colistimethate for Injection

(Comment on this Monograph)id=m19888=Colistimethate for Injection=Chl-Cy-Monos.pdf) DEFINITION Colistimethate for Injection contains an amount of Colistimethate Sodium equivalent to NLT 90.0% and NMT 120.0% of the labeled amount of colistin. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample solution A: (where it is represented as being in a single-dose container): Constitute Colistimethate for Injection in a volume of water corresponding to the volume of diluent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with Buffer No. 6 to obtain a solution having a convenient concentration. Sample solution B: (where the label states the quantity of colistin equivalent in a given volume of constituted solution): Constitute Colistimethate for Injection in a volume of water corresponding to the volume of diluent specified in the labeling. Dilute a measured volume of the constituted solution with Buffer No. 6 to obtain a solution having a convenient concentration. Analysis: Proceed as directed for AntibioticsMicrobial Assays 81, Colistimethate Sodium, using a measured volume of Sample solution diluted with Buffer No. 6 to yield a Sample Dilution having a concentration assumed to be equal to the median dose level of the Standard. Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231:
Colistin, sulfate. Colistins sulfate [1264-72-8]. DEFINITION Colistin Sulfate is the sulfate salt of an antibacterial substance produced by the growth of Bacillus polymyxa var. colistinus. It has a potency equivalent to NLT 500 g of colistin /mg. IDENTIFICATION A. PROCEDURE Solution A: Adjust 50 mL of 1 M monobasic potassium phosphate with 1 N sodium hydroxide to a pH of 7.0, and dilute with water to 100 mL. Sample solution: Dissolve 20 mg of colistin sulfate in 2 mL of Solution A. Analysis: Add 0.2 mL of 5 mg/mL ninhydrin solution to the Sample solution, and boil. Acceptance criteria: A purple color is produced. B. IDENTIFICATION TESTSGENERAL, Sulfate 191 C. LIQUID CHROMATOGRAPHIC IDENTIFICATION TEST Mobile phase: Acetonitrile and 0.1 M tribasic sodium phosphate (23:77) Adjust with phosphoric acid to a pH of 3.0. Standard solution: 6 mg/mL of USP Colistin Sulfate RS in Mobile phase [NOTEProtect this solution from light.] Sample solution: 6 mg/mL of Colistin Sulfate in Mobile phase [NOTEProtect this solution from light.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 212 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 10 L Analysis Samples: Standard solution and Sample solution [NOTEThe relative retention times for colistin A and colistin B are about 0.8 and 0.55, respectively.] Acceptance criteria: The chromatogram of the Sample solution corresponds qualitatively with that of the Standard solution. ASSAY PROCEDURE Analysis: Proceed as directed for Colistin under Antibiotics Microbial Assays 81.
NMT 0.003%
SPECIFIC TESTS FREE COLISTIN Sample solution: Dissolve 80 mg in 3 mL of water, and add 0.05 mL of 100 mg/mL silicotungstic acid solution. Acceptance criteria: No immediate precipitate is formed. BACTERIAL ENDOTOXINS TEST 85: Contains NMT 2.0 USP Endotoxin Units per mg of colistin STERILITY TESTS 71: Meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration INJECTIONS 1: At the time of use, it meets the requirements for Injections 1, Constituted Solutions and for Injections 1, Labeling. PH 791: 6.58.5, in 10 mg/mL solution LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 7.0% of its weight.
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NLT 500 g/mg
USP 32
amount of hydrocortisone acetate (C23H32O6). It contains one or more suitable buffers, detergents, dispersants, and preservatives. [NOTEWhere Colistin and Neomycin Sulfates and Hydrocortisone Acetate Otic Suspension is prescribed, without reference to the quantity of colistin, neomycin, or hydrocortisone acetate contained therein, a product containing 3.0 mg of colistin, 3.3 mg of neomycin, and 10 mg of hydrocortisone acetate/mL shall be dispensed.] ASSAY COLISTIN Sample solution: Use a freshly mixed, measured volume of Otic Suspension diluted with Buffer No. 6 to yield a sample dilution having a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81. Acceptance criteria: 90.0%135.0% NEOMYCIN Sample solution: Use a freshly mixed, measured volume of Otic Suspension diluted with Buffer No. 3 to yield a sample dilution having a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81. Acceptance criteria: 90.0%125.0% HYDROCORTISONE ACETATE Reagent blank: 22 N sulfuric acid and dehydrated alcohol (2:1) Reagent A: 433.3 g/mL of phenylhydrazine hydrochloride in Reagent blank Standard solution: 10 g/mL of USP Hydrocortisone Acetate RS in chloroform Sample solution: Transfer 5.0 mL of freshly mixed Otic Suspension to a 125-mL separator. Extract with three 20-mL portions of chloroform, filtering each chloroform extract through a pledget of cotton previously saturated with chloroform, collect the filtrates in a 100-mL volumetric flask, and dilute with chloroform to volume. Dilute 10 mL of the resulting solution with chloroform to 100-mL, and dilute 20 mL of this solution with chloroform to 100-mL. Analysis: Pipet 50 mL each of the Standard solution and the Sample solution into separate 125-mL separators, add 2 mL of 0.1 N sodium hydroxide to each separator, shake, and allow the layers to separate. Filter both chloroform layers through glass wool, and collect the filtrates in separate beakers. Pipet two 20-mL portions of each chloroform filtrate into separate 125-mL separators. Add 25.0 mL of Reagent A to one separator of each of the filtrates from the Standard solution and the Sample solution, respectively, and add 25.0 mL of Reagent blank to the remaining two separators. Shake all four separators well, allow the layers to separate, and discard the chloroform layers. Drain the aqueous layers into separate centrifuge tubes, and centrifuge for 2 min. Pipet 10 mL of each solution into separate glassstoppered test tubes. Place the tubes in a water bath maintained at a temperature of 60 for 30 min, then cool the solution to room temperature. Spectrometric conditions Mode: Spectrometry Analytical wavelength: 410 nm Blank: Water Analysis Samples: Reagent blank, Standard solutions, and Sample solutions Calculate the percentage of C23H32O6 in the Otic Suspension taken: Result = (AUAub)/(ASAsb) CS 100
SPECIFIC TESTS PH 791: 4.07.0, in 10 mg/mL solution LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 7.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Colistin Sulfate RS
Colistin Sulfate for Oral Suspension

(Comment on this Monograph)id=m19930=Colistin Sulfate for Oral Suspension=Chl-Cy-Monos.pdf) DEFINITION Colistin Sulfate for Oral Suspension is a dry mixture of Colistin Sulfate with or without one or more suitable buffers, colors, diluents, dispersants, and flavors. It contains the equivalent of NLT 90.0% and NMT 120.0% of the labeled amount of colistin. ASSAY PROCEDURE Sample solution: Constitute Colistin Sulfate for Oral Suspension as directed in the labeling. Use a measured volume of this suspension, freshly mixed and free from air bubbles, diluted with Buffer No. 6 to yield a sample dilution having a concentration assumed to be equal to the median dose level of the Standard. Analysis: Proceed as directed under AntibioticsMicrobial Assays 81. Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for solids packaged in single-unit containers DELIVERABLE VOLUME 698: Meets the requirements SPECIFIC TESTS PH 791: 5.06.0, in the suspension constituted as directed in the labeling LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 3.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. USP REFERENCE STANDARDS 11 USP Colistin Sulfate RS
Colistin and Neomycin Sulfates and Hydrocortisone Acetate Otic Suspension

(Comment on this Monograph)id=m19950=Colistin and Neomycin Sulfates and Hydrocortisone Acetate Otic Suspension=Chl-Cy-Monos.pdf) DEFINITION Colistin and Neomycin Sulfates and Hydrocortisone Acetate Otic Suspension is a sterile suspension containing the equivalent of NLT 90.0% and NMT 135.0% of the labeled amount of colistin, NLT 90.0% and NMT 125.0% of the labeled amount of neomycin, and NLT 90.0% and NMT 110.0% of the labeled
USP 32
= absorbance of the Sample solution treated with Reagent A = absorbance of the Sample solution treated with Aub the Reagent blank = absorbance of the Standard solution treated with AS Reagent A = absorbance of the Standard solution treated with Asb the Reagent blank = concentration of the USP Hydrochloride Acetate CS RS in the Standard solution (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.85.2 STERILITY TESTS 71: Meets the requirements, 0.25 mL from each container being transferred directly to 90 mL of each medium ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Colistin Sulfate RS USP Hydrocortisone Acetate RS USP Neomycin Sulfate RS AU
Official Monographs / Collodion 185

Standard solutions: Mix 10 mL of each Standard stock solution with 15 mL of 1,2-dichloroethane, add 10 mL of hexane, and 10.0 mL of Internal standard solution in separate glass-stoppered 50-mL graduated cylinders. Sample solution: Mix 10 mL of Collodion with 15 mL of 1,2-dichloroethane in a glass-stoppered 50-mL graduate cylinder with 10 mL of hexane, and 10.0 mL of Internal standard solution. Mix and allow the precipitate to settle. Chromatographic system Mode: GC Detector: Thermal conductivity Column: 1.8-m 3.5-mm glass column containing packing S3 Temperature Column: 150 Injector: 200 Detector: 250 Carrier gas: Helium Flow rate: 50 mL/min Injection size: 4 L Analysis Samples: Standard solutions and Sample solution Calculate the relative response factor, F, for each Standard solution taken: F = CS/RS CS = concentration of C2H5OH in the Standard solution, in percentage = area ratio of alcohol to acetone observed for the RS respective Standard solution Calculate the percentage of alcohol, in the portion of Collodion taken: Result = Fa RU = average of the individual F values = area ratio of alcohol to acetone observed for the Sample solution Acceptance criteria: 22.0%26.0% SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.7650.775 ACIDITY Sample: 5 mL Analysis: Add 5 mL of water. Acceptance criteria: The liquid separated from the pyroxylin is not acid to litmus. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at a temperature not exceeding 30, remote from fire. LABELING: The label bears a caution statement to the effect that Collodion is highly flammable. Fa RU
Collodion
(Comment on this Monograph)id=m20010=Collodion=Chl-CyMonos.pdf) DEFINITION Collodion contains NLT 5.0% by weight, of pyroxylin.
Pyroxylin Ether Alcohol To make about 40 g 750 mL 250 mL 1000 mL
Add the Alcohol and the Ether to the Pyroxylin contained in a suitable container, and insert the stopper into the container well. Shake the mixture occasionally until the Pyroxylin is dissolved. [CAUTIONCollodion is highly flammable.] IDENTIFICATION A. When exposed to air in a thin layer, it leaves a transparent, tenacious film. The film of pyroxylin so obtained burns rapidly, with a yellow flame. B. When mixed with an equal volume of water, it yields a viscid, stringy mass of pyroxylin. ASSAY PROCEDURE Analysis: Pour quickly 10 mL of Collodion into a tared flask, insert the stopper, weigh the assay charge accurately, remove the stopper, warm on a steam bath, and add 10 mL of water dropwise, with constant stirring. Evaporate the mixture on a steam bath, and dry the residue at 105 for 4 h. Acceptance criteria: NLT 5.0%, by weight, of pyroxylin OTHER COMPONENTS ALCOHOL DETERMINATION 611 Internal standard solution: Acetone and 1,2dichloroethane (1:4) Standard stock solutions: Dilute 10-, 20-, and 30-mL portions of dehydrated alcohol in separate 100-mL volumetric flasks with 1,2-dichloroethane.
Flexible Collodion
(Comment on this Monograph)id=m20020=Flexible Collodion=Chl-Cy-Monos.pdf) DEFINITION Prepare Flexible Collodion as follows:
Camphor Castor Oil Collodion To make 20 g 30 g A sufficient quantity 1000 g
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Collodion / Official Monographs
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Weigh the ingredients, successively, into a dry, tared bottle, insert the stopper in the bottle, and shake the mixture until the camphor is dissolved. IDENTIFICATION A. When exposed to air in a thin layer, it leaves a transparent, tenacious film. The film exhibits a distinct odor of camphor. The film of pyroxylin so obtained burns rapidly, with a yellow flame. B. When mixed with an equal volume of water, it yields a viscid, stringy mass of pyroxylin. OTHER COMPONENTS ALCOHOL DETERMINATION 611 Internal standard solution: Acetone and 1,2dichloroethane (1:4) Standard stock solutions: Dilute 10-, 20-, and 30-mL portions of dehydrated alcohol in separate 100-mL volumetric flasks with 1,2-dichloroethane. Standard solutions: Mix 10 mL of each Standard stock solution with 15 mL of 1,2-dichloroethane, add 10 mL of hexane, and 10.0 mL of Internal standard solution in separate glass-stoppered 50-mL graduated cylinders. Sample solution: Mix 10 mL of Flexible Collodion in a 50mL glass-stoppered graduated cylinder with 15 mL of 1,2dichloroethane, add 10 mL of hexane, and 10.0 mL of Internal standard solution. Mix and allow the precipitate to settle. Chromatographic system Mode: GC Detector: Thermal conductivity Column: 1.8-m 3.5-mm glass column containing packing S3 Temperature Column: 150 Injector: 200 Detector: 250 Carrier gas: Helium Flow rate: 50 mL/min Injection size: 4 L Analysis Samples: Standard solutions and Sample solution Calculate the relative response factor, F, for each Standard solution taken: F = CS/RS CS = concentration of C2H5OH in the Standard solution, in percentage = area ratio of alcohol to acetone observed for the RS respective Standard solution Calculate the percentage of alcohol in Flexible Collodion taken: Result = Fa RU = average of the individual F values = area ratio of alcohol to acetone observed for the Sample solution Acceptance criteria: 21.0%25.0% Fa RU SPECIFIC TESTS SPECIFIC GRAVITY 841: 0.7700.790
Colloidal Oatmeal
(Comment on this Monograph)id=m20023=Colloidal Oatmeal=Chl-Cy-Monos.pdf) DEFINITION Colloidal Oatmeal is the powder resulting from the grinding and further processing of whole oat grain meeting U.S. Standards for Number 1 or Number 2 oats (7 CFR 810.1001). IDENTIFICATION A. PROCEDURE Analysis: Prepare a smooth mixture of 10 g of Colloidal Oatmeal and 100 mL of warm water. Acceptance criteria: After stirring for 10 min, the resulting slurry has a characteristic slippery feel and shows the development of slimy, viscous strands. B. A water slurry is colored reddish violet to deep blue by iodine TS. OTHER COMPONENTS NITROGEN DETERMINATION, Method I 461: NLT 2.0%
IMPURITIES Inorganic Impurities ARTICLES OF BOTANICAL ORIGIN, Total Ash 561: calculated on the dried basis
NMT 2.5%,
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, at a temperature not exceeding 30, remote from fire. LABELING: The label bears a caution statement to the effect that Flexible Collodion is highly flammable.
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: The total aerobic microbial count does not exceed 10,000 cfu/g; the total combined molds and yeasts count does not exceed 150 cfu/g. LOSS ON DRYING 731: Dry it at 120 for 4 h; it loses NMT 10% of its weight. PARTICLE SIZE DISTRIBUTION ESTIMATION BY ANALYTICAL SIEVING 786: NMT 3% of the total particles exceed 150 m in size and NMT 20% of the total particles exceed 75 m in size. FAT CONTENT Analysis: Dry 4 g in vacuum at 100 for about 5 h. Extract the fat from it with anhydrous ethyl ether, using a continuous extraction apparatus, the extraction period being 4 h at a condensation rate of 5 to 6 drops/s. Evaporate the ether from the extract, transfer it to a tared beaker, and dry at 100 to constant weight. Perform a blank determination. Calculate the percentage of fat found, corrected for the blank. Acceptance criteria: NLT 0.2% VISCOSITY 911 Sample solution: Transfer 25 g of Colloidal Oatmeal in small portions, with stirring at 1000 rpm over a 1-min period, to 500 mL of water contained in a beaker, maintained at 45 and equipped with a variable speed mixer. Stir for 5 min after the addition of the last portion of oatmeal. Allow the suspension to stand for 90 min, and equilibrate to ambient temperature. Stir the suspension at 800 rpm for 1 min. Apparatus: Equip a suitable rotational viscosimeter with a spindle having a cylinder 1.88 cm in diameter and 6.25 cm high attached to a shaft 0.32 cm in diameter, the distance from the top of the cylinder to the lower tip of the shaft being 0.75 cm, and the immersion depth being 8.15 cm (No. 1 spindle). Analysis: Determine and record the viscosity of the suspension, with the spindle rotating at 60 rpm. Convert to centipoise by multiplying the reading by the constant for the viscosimeter spindle and speed employed.
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Acceptance criteria: The average of three viscosities obtained is greater than 1 and less than 100 centipoises. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers.
Official Monographs / Copper 187

CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion dissolved in boiling water shows no more sulfate than corresponds to 1 mL of 0.020 N sulfuric acid (0.05%). ARSENIC, Method I 211: NMT 3 ppm Dissolve 1.0 g in 35 mL of water. LEAD 251: [NOTEFor the preparation of all aqueous solutions and for the rinsing of glassware before use, use water that has been passed through a strong-acid, strong-base, mixed-bed ion-exchange resin. Select all reagents to have as low a content of lead as practicable, and store all reagent solutions in containers of borosilicate glass. Cleanse glassware before use by soaking in warm 8 N nitric acid for 30 min and by rinsing with deionized water.] Standard solution: Transfer 10.0 mL of Lead Nitrate Stock Solution, prepared as directed in the test for Heavy Metals 231, to a 100-mL volumetric flask, add 40 mL of water and 5 mL of nitric acid, and dilute with water to volume. Mix 0.40 mL of this solution with 50 mL of water, add 1 mL of nitric acid, and dilute with water to 100 mL. This solution contains 0.04 g/mL of lead. Sample stock solution: Transfer 4 g of Copper Gluconate, to a 100-mL volumetric flask. Add 50 mL of water and 5 mL of nitric acid, and sonicate to dissolve the specimen. Dilute with water to volume. Transfer 4.0 mL of this solution to a second 100-mL volumetric flask, add 50 mL of water, and 1 mL of nitric acid, dilute with water to volume, and mix. Blank: Water and nitric acid (98.8:1.2) Sample solutions: Prepare mixtures of the Sample stock solution, the Standard solution, and the Blank with the following proportional compositions, by volume: 10.0:0:10.0, 10.0:4.0:6.0, 10.0:7.0:3.0, and 10.0:10.0:0. These Sample solutions contain 0, 0.008, 0.014, and 0.020 g/mL of lead from the Standard solution, respectively. Spectrophotometric conditions Mode: Spectrophotometry Injection volume: 20 L Temperature: See the Temperature Program Table below.
Time (s) 10 60 15 5 10 2 3.2 Temperature () 70 90 120 250 (no gas flow) 250 250 (no gas flow) 2000
Copper Gluconate
(Comment on this Monograph)id=m20040=Copper Gluconate=Chl-Cy-Monos.pdf)
C12H22CuO14 Copper, bis(D-gluconato-O1,O2)-; Copper D-gluconate (1:2) [527-09-3].
453.84
DEFINITION Copper Gluconate contains NLT 98.0% and NMT 102.0% of C12H22CuO14. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Copper 191: 50 mg/mL B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 10 mg/mL of USP Potassium Gluconate RS Heat in a water bath at 60 if necessary. Sample solution: 10 mg/mL Heat in a water bath at 60 if necessary. Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 5 L Developing solvent system: Alcohol, ethyl acetate, ammonium hydroxide, and water (5:1:1:3) Spray reagent: Dissolve 2.5 g of ammonium molybdate in 50 mL of 2 N sulfuric acid, add 1.0 g of ceric sulfate, swirl to dissolve, and dilute with 2 N sulfuric acid to 100 mL. Analysis Samples: Standard solution and Sample solution Analysis: Proceed as directed for Chromatography 621 Thin-Layer Chromatography. Develop the chromatogram in the Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, and dry at 110 for 20 min. Allow to cool, spray with the Spray reagent. Heat the plate at 110 for 10 min. Acceptance criteria: The principal spot from the Sample solution corresponds in color, size, and RF value to that from the Standard solution. ASSAY PROCEDURE Sample: 1.5 g Analysis: Dissolve the Sample in 100 mL of water. Add 2 mL of glacial acetic acid and 5 g of potassium iodide. Titrate with 0.1 N sodium thiosulfate VS to a light yellow color. Add 2 g of ammonium thiocyanate, mix, add 3 mL of starch TS, and continue titrating to a milk-white endpoint. Each mL of 0.1 N sodium thiosulfate is equivalent to 45.38 mg of C12H22CuO14. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities CHLORIDE AND SULFATE, Chloride 221: A 1.0-g portion shows no more chloride than corresponds to 1 mL of 0.020 N hydrochloric acid (0.07%).
Argon gas flow: 3 L/min Analysis Samples: Sample solutions and the Blank Inject the solutions into the graphite tube of a suitable graphite furnace atomic absorption spectrophotometer, temperature-programmed to reach 2000 in about 2 min as shown in the Temperature Program Table above. When the temperature reaches 2000, determine the absorbance at the lead emission line at 283.3 nm, corrected for background absorption. Correct the absorbance values from the Sample solutions by subtracting from each the absorbance value from the Blank. Plot the corrected absorbances of the Sample solutions versus their contents of lead, in g/mL, as furnished by the Standard solution, draw the straight line best fitting the four points, and extrapolate the line until it intercepts the concentration axis. From the intercept, determine the concentration, C, in g/mL, of lead in each mL of the Sample solution containing 0 g of lead from the Sample stock solution.
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Calculate the percentage of lead in the specimen taken: Result = (C/W) D F 100
USP 32
restrict the selection so that no rat is more than 30% heavier than the lightest, and the number of rats is an exact multiple of six. Separate the selected rats into six groups, equal in size, of NLT six rats each, and assign at random one of the three diluted Standard solutions or one of the three Sample solutions to each group. Analysis: Inject all rats of each group subcutaneously with the assigned test doses. Three h after the injection, anesthetize the rats, and remove both adrenal glands from each rat, free them from adhering tissue, and promptly weigh each pair on a suitable balance to the nearest 0.2 mg. Place the weighed glands from each rat in suitable vessels each containing 8.0 mL of metaphosphoric acid solution (1 in 40), and comminute the glands by grinding with a small quantity of washed sand. Cover each vessel, and proceed similarly until all glands have been extracted. Ascorbic acid determination: Filter the metaphosphoric acid extracts, and pipet 4 mL of each filtrate into suitable vessels each containing 4.0 mL of indophenol-acetate TS. Mix by shaking, and read the absorbance at 520 nm, with a suitable spectrophotometer. From the observed absorbance and the standard curve prepared as directed in the next paragraph, calculate the amount of ascorbic acid in terms of mg of ascorbic acid in each 100 g of adrenal gland tissue. Prepare a standard concentrationabsorbance curve, using three standard solutions containing in each mL, respectively, 6.0, 8.0, and 10.0 g of USP Ascorbic Acid RS in metaphosphoric acid solution (1 in 40). Pipet into each of three suitable vessels, preferably spectrophotometer cells, 4 mL of indophenol-acetate TS. Add 4.0 mL of one of the three standard ascorbic acid solutions to one of the cells, mix, and promptly read the absorbance in the same instrument and under the same conditions as for the adrenal gland extracts. Repeat the process for the other two standard ascorbic acid solutions, plot the concentrationabsorbance values, and draw the straight line best fitting the three plotted points. Calculation: Tabulate the observed concentration of ascorbic acid in the adrenal glands of each rat, designated by the symbol y, for each dosage group of f rats. If the data from one or more rats are missing, adjust to groups of equal size by suitable means (see Design and Analysis of Biological Assays 111, Replacement of Missing Values). Total the values of y in each group, and designate each total as T, subscripts 13 for the three successive dosage levels and subscripts S and U for the Standard and the Injection, respectively. Test both the agreement in slope of the dosage-response lines for the Standard and for the Injection, and the lack of curvature, as directed for a 3-dose balanced assay (see Design and Analysis of Biological Assays 111, Tests of Assay Validity). If the combined discrepancy as measured by F3 exceeds its tabular value in Table 9 (see Design and Analysis of Biological Assays 111, Combination of Independent Assays), regard these data as preliminary and repeat the assay. Determine the logarithm of potency of the Injection taken: M = (4iTa/3Tb) + log R = interval between successive log doses of both the Standard solution and the Sample solution = (TU TS) Ta = (T3 T1) Tb R = vS/vU, the ratio of the high dose of the Standard in USP Units (vS) to the high dose of the Injection in mL (vU) Compute the log confidence interval L (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). Replication: Repeat the entire determination at least once. Test the agreement among the two or more independent i
= measured concentration of lead in the Sample solution (g/mL) W = weight of Copper Gluconate taken to prepare the Sample stock solution (g) D = dilution volume of the Sample stock solution, (5000 mL) F = unit conversion factor, 106 (g/g) Acceptance criteria: NMT 25ppm SPECIFIC TESTS REDUCING SUBSTANCES Sample solution: Transfer 1.0 g to a 250-mL conical flask, dissolve in 10 mL of water, and add 25 mL of alkaline cupric citrate TS. Cover the flask, boil gently for 5 min, and cool rapidly to room temperature. Add 25 mL of 0.6 N acetic acid, 10.0 mL of 0.1 N iodine VS, and 10 mL of 3 N hydrochloric acid. Analysis: Titrate with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as the endpoint is approached. Perform a blank determination, omitting the specimen, and note the difference in volumes required (see Titrimetry 541). Each mL of the difference in volume of 0.1 N sodium thiosulfate consumed is equivalent to 2.7 mg of reducing substances (as dextrose). Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Potassium Gluconate RS
Corticotropin Injection
(Comment on this Monograph)id=m20150=Corticotropin Injection=Chl-Cy-Monos.pdf) Corticotropin [9002-60-2]. DEFINITION Corticotropin Injection is a sterile solution, in a suitable diluent, of the material containing the polypeptide hormone having the property of increasing the rate of secretion of adrenal corticosteroids, which is of the anterior lobe of the pituitary of mammals used for food by man. Its potency is NLT 80.0% and NMT 125.0% of the potency stated on the label in USP Corticotropin Units. It may contain a suitable antimicrobial agent. ASSAY PROCEDURE Standard solution: Pipet 2.5 mL of gelatin TS into an opened container of USP Corticotropin RS, and mix, to obtain a solution having a concentration of 2.0 USP Corticotropin Units/mL. Using gelatin TS as a diluent, prepare three diluted standard solutions such that the respective concentrations of corticotropin constitute a geometric series such as 1:2:4 or 1:3:9 and such that the quantity of corticotropin in each 0.5 mL lies within the range of 10300 milliunits. Sample solution: In the same manner, using the same diluent, dilute the Injection to give three Sample solutions corresponding to those of the standard. The animals: Select healthy rats, of the same or either sex, that have been raised on a diet fully adequate with respect to vitamin and mineral content. Anesthetize the rats with ether, and remove the hypophysis from each by application of gentle suction through a fine-tipped tube. 1648 h after the operation, select those rats weighing 80180 g, but
USP 32
determinations, and compute the weight for each (see Design and Analysis of Biological Assays 111, Combination of Independent Assays). Calculate the weighted mean logpotency M and its confidence interval, Lc (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). The potency, P*, is satisfactory if P* = antilog M is 80%125% of the labeled potency and if the confidence interval does not exceed 0.40. Acceptance criteria: 80.0%125.0% IMPURITIES Organic Impurities VASOPRESSIN ACTIVITY Solution A: Dissolve 6.6 g of dibasic ammonium phosphate in 950 mL of water, and adjust with concentrated phosphoric acid to a pH of 3.0. Dilute with water to 1 L. Mobile phase: Acetonitrile and Solution A (13:87) [NOTEThe retention time of the vasopressin peak is very sensitive to small changes in the acetonitrile concentration.] Standard solution: Dissolve the entire contents of a vial of USP Vasopressin RS in a known volume of Solution A, and dilute with Solution A to obtain a final solution containing 0.1 USP Vasopressin Unit/mL. Sample solution: Dissolve the entire contents of a vial of the Injection in a known volume of Solution A, and dilute with Solution A to obtain a final solution containing 2.0 USP Corticotropin Units/mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6 mm 25 cm; 5 m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the vasopressin activity in USP Vasopressin Unit /USP Corticotropin Unit taken: Result = (rU/rS) 2 CS = peak response from the Sample solution = peak response from the Standard solution = concentration of Standard solution (USP Vasopressin Units/mL) Acceptance criteria: NMT 0.05 rU rS CS SPECIFIC TESTS PH 791: 3.07.0 PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections. BACTERIAL ENDOTOXINS TEST 85: Contains NMT 3.1 USP Endotoxin Units/USP Corticotropin Unit. OTHER REQUIREMENTS: Meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. Store in a cold place. LABELING: If the labeling of Injection recommends intravenous administration, include specific information on dosage. USP REFERENCE STANDARDS 11 USP Ascorbic Acid RS USP Corticotropin RS
Official Monographs / Corticotropin 189

USP Endotoxin RS USP Vasopressin RS
Corticotropin for Injection

(Comment on this Monograph)id=m20160=Corticotropin for Injection=Chl-Cy-Monos.pdf) Corticotropin [9002-60-2]. DEFINITION Corticotropin for Injection is the sterile, dry material containing the polypeptide hormone having the property of increasing the rate of secretion of adrenal corticosteroids, which is obtained from the anterior lobe of the pituitary of mammals used for food by man. Its potency is NLT 80.0% and NMT 125.0% of the potency stated on the label in USP Corticotropin Units. It may contain a suitable antimicrobial agent and suitable diluents and buffers. ASSAY PROCEDURE Standard solutions: Pipet 2.5 mL of gelatin TS into an opened container of USP Corticotropin RS, and mix, to obtain a solution having a concentration of 2.0 USP Corticotropin Units/mL. Using gelatin TS as a diluent, prepare three diluted standard solutions such that the respective concentrations of corticotropin constitute a geometric series such as 1:2:4 or 1:3:9, and such that the quantity of corticotropin in each 0.5 mL lies within the range of 10 to 300 milliunits. Sample solutions: Constitute the Corticotropin for Injection as directed in the labeling. In the same manner as the Standard solutions preparation, using the same diluent, dilute the Corticotropin for Injection to give three Sample solutions corresponding to those of the Standard solutions. The animals: Select healthy rats, of the same or either sex, that have been raised on a diet fully adequate with respect to vitamin and mineral content. Anesthetize the rats with ether, and remove the hypophysis from each by application of gentle suction through a fine-tipped tube. Between 16 and 48 h after the operation, select those rats weighing between 80 and 180 g, but restrict the selection so that no rat is more than 30% heavier than the lightest, and the number of rats is an exact multiple of 6. Separate the selected rats into 6 groups, equal in size, of NLT 6 rats each, and assign at random one of the three diluted Standard solutions or of the three Sample solutions to each group. Analysis: Inject all rats of each group subcutaneously with the assigned test doses. Three h after the injection, anesthetize the rats, and remove both adrenal glands from each rat, free them from adhering tissue, and promptly weigh each pair on a suitable balance to the nearest 0.2 mg. Place the weighed glands from each rat in suitable vessels each containing 8.0 mL of metaphosphoric acid solution (1 in 40), and comminute the glands by grinding with a small quantity of washed sand. Cover each vessel, and proceed similarly until all glands have been extracted. Ascorbic acid determination: Filter the metaphosphoric acid extracts, and pipet 4 mL of each filtrate into suitable vessels each containing 4.0 mL of indophenol-acetate TS. Mix by shaking, and read the absorbance at 520 nm, with a suitable spectrophotometer. From the observed absorbance and the standard curve prepared as directed in the next paragraph, calculate the amount of ascorbic acid in terms of mg of ascorbic acid in each 100 g of adrenal gland tissue. Prepare a standard concentration-absorbance curve, using three standard solutions containing in each mL, respectively, 6.0, 8.0, and 10.0 g of USP Ascorbic Acid RS in metaphosphoric acid solution (1 in 40). Pipet into each of three suitable vessels, preferably spectrophotometer cells,
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Sample solution: Dissolve the entire contents of a vial of Corticotropin for Injection in a known volume of Solution A, and dilute with Solution A to obtain a final solution containing 2.0 USP Corticotropin Units/mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the vasopressin activity in USP Vasopressin Unit/USP Corticotropin Unit: Result = (rU/rS)/2 CS = peak response from the Sample solution = peak response from the Standard solution = concentration of the Standard solution (units/mL) Acceptance criteria: NMT 0.05 rU rS CS SPECIFIC TESTS PH 791: 2.56.0, in a solution constituted as directed in the labeling supplied by the manufacturer PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections Sterility Tests 71 Meets the requirements BACTERIAL ENDOTOXINS TEST 85: Contains NMT 3.1 USP Endotoxin Units/USP Corticotropin Unit. OTHER REQUIREMENTS: Meets the requirements for Injections 1, Constituted Solutions and Labels and Labeling. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described under Injections 1, Containers for Sterile Solids. LABELING: If the labeling of Corticotropin for Injection recommends intravenous administration, include specific information on dosage. USP REFERENCE STANDARDS 11 USP Ascorbic Acid RS USP Corticotropin RS USP Endotoxin RS USP Vasopressin RS
4 mL of indophenolacetate TS. Add 4.0 mL of one of the three standard ascorbic acid solutions to one of the cells, mix, and promptly read the absorbance in the same instrument, and under the same conditions as for the adrenal gland extracts. Repeat the process for the other two standard ascorbic acid solutions, plot the concentration-absorbance values, and draw the straight line best fitting the three plotted points. Calculation: Tabulate the observed concentration of ascorbic acid in the adrenal glands of each rat, designated by the symbol y, for each dosage group of f rats. If the data from one or more rats are missing, adjust to groups of equal size by suitable means (see Design and Analysis of Biological Assays 111, Replacement of Missing Values). Total the values of y in each group, and designate each total as T, subscripts 1 to 3 for the three successive dosage levels and subscripts S and U for the Standard and the Corticotropin for Injection, respectively. Test both the agreement in slope of the dosage-response lines for the Standard and for the Corticotropin for Injection, and the lack of curvature, as directed for a 3-dose balanced assay (see Design and Analysis of Biological Assays 111, Experimental Error and Tests of Assay Validity ). If the combined discrepancy as measured by F3 exceeds its tabular value in Table 9 (see Design and Analysis of Biological Assays 111, Combination of Independent Assays), regard these data as preliminary and repeat the Assay. Determine the logarithm of potency of the Injection: M = (4iTa/3Tb) + log R = interval between successive log doses of both the Standard solution and the Sample solution = (TU TS) Ta Tb = (T3 T1) R = vS/vU, the ratio of the high dose of the Standard in USP Units (vS) to the high dose of the Injection in mL (vU) Compute the log confidence interval L (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). Replication: Repeat the entire determination at least once. Test the agreement among the two or more independent determinations, and compute the weight for each (see Design and Analysis of Biological Assays 111, Combination of Independent Assays). Calculate the weighted mean logpotency M and its confidence interval, Lc (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). The potency, P, is satisfactory if P = antilog M is 80% 125% of the labeled potency and if the confidence interval does not exceed 0.40. Acceptance criteria: 80.0%125.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements i
Repository Corticotropin Injection

(Comment on this Monograph)id=m20170=Repository Corticotropin Injection=Chl-Cy-Monos.pdf) Corticotropin [9002-60-2]. DEFINITION Repository Corticotropin Injection is corticotropin in a solution of partially hydrolyzed gelatin. Its potency is NLT 80.0% and NMT 125.0% of the potency stated on the label in USP Corticotropin Units. It may contain a suitable antimicrobial agent. ASSAY PROCEDURE Standard solution: Pipet 2.5 mL of gelatin TS into an opened container of USP Corticotropin RS, and mix, to obtain a solution having a concentration of 2.0 USP
IMPURITIES Organic Impurities VASOPRESSIN ACTIVITY Solution A: Dissolve 6.6 g of dibasic ammonium phosphate in 950 mL of water, and adjust with concentrated phosphoric acid to a pH of 3.0. Dilute with water to 1 L. Mobile phase: Acetonitrile and Solution A (13:87) [NOTEThe retention time of the vasopressin peak is very sensitive to small changes in the acetonitrile concentration.] Standard solution: Dissolve the entire contents of a vial of USP Vasopressin RS in a known volume of Solution A, and dilute with Solution A to obtain a final solution containing 0.1 USP Vasopressin Unit/mL.
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Corticotropin Units/mL. Using gelatin TS as a diluent, prepare three diluted Standard solutions such that the respective concentrations of corticotropin constitute a geometric series such as 1:2:4 or 1:3:9, and such that the quantity of corticotropin in each 0.5 mL lies within the range of 10 300 milliunits. Sample solution: In the same manner, using the same diluent, dilute the Injection to give three test solutions corresponding to those of the standard. The animals: Select healthy rats, of the same or either sex, that have been raised on a diet fully adequate with respect to vitamin and mineral content. Anesthetize the rats with ether, and remove the hypophysis from each by application of gentle suction through a fine-tipped tube. 16 48 h after the operation, select those rats weighing 80 180 g, but restrict the selection so that no rat is more than 30% heavier than the lightest, and the number of rats is an exact multiple of six. Separate the selected rats into six groups, equal in size, of NLT six rats each, and assign at random one of the three diluted standard solutions or of the three test solutions to each group. Analysis: Inject all rats of each group subcutaneously with the assigned test doses. Three h after the injection, anesthetize the rats, remove both adrenal glands from each rat, free them from adhering tissue, and promptly weigh each pair on a suitable balance to the nearest 0.2 mg. Place the weighed glands from each rat in suitable vessels each containing 8.0 mL of metaphosphoric acid solution (1 in 40), and comminute the glands by grinding with a small quantity of washed sand. Cover each vessel, and proceed similarly until all glands have been extracted. Ascorbic acid determination: Filter the metaphosphoric acid extracts, and pipet 4 mL of each filtrate into suitable vessels each containing 4.0 mL of indophenol-acetate TS. Mix by shaking, and read the absorbance at 520 nm, with a suitable spectrophotometer. From the observed absorbance and the standard curve prepared as directed in the next paragraph, calculate the amount, in mg, of ascorbic acid in each 100 g of adrenal gland tissue. Prepare a standard concentration-absorbance curve, using 3 standard solutions containing in each mL, respectively, 6.0, 8.0, and 10.0 g of USP Ascorbic Acid RS in metaphosphoric acid solution (1 in 40). Pipet into each of 3 suitable vessels, preferably spectrophotometer cells, 4 mL of indophenol-acetate TS. Add 4.0 mL of one of the 3 standard ascorbic acid solutions to one of the cells, mix, and promptly read the absorbance in the same instrument and under the same conditions as for the adrenal gland extracts. Repeat the process for the other two standard ascorbic acid solutions, plot the concentration-absorbance values, and draw the straight line best fitting the 3 plotted points. Calculation: Tabulate the observed concentration of ascorbic acid in the adrenal glands of each rat, designated by the symbol y, for each dosage group of f rats. If the data from one or more rats are missing, adjust to groups of equal size by suitable means (see Design and Analysis of Biological Assays 111, Replacement of Missing Values). Total the values of y in each group, and designate each total as T, subscripts 1 to 3 for the three successive dosage levels and subscripts S and U for the Standard and the Injection, respectively. Test both the agreement in slope of the dosage-response lines for the Standard and for the Injection, and the lack of curvature, as directed for a 3-dose balanced assay (see Design and Analysis of Biological Assays 111, Experimental Error and Tests of Assay Validity). If the combined discrepancy as measured by F3 exceeds its tabular value in Table 9 (see Design and Analysis of Biological Assays 111, Combination of Independent Assays), regard these data as preliminary and repeat the assay.
Official Monographs / Corticotropin 191

Determine the logarithm of potency (M) of the Injection: M = (4iTa/3Tb)+ log R = interval between successive log doses of both the Standard solution and the Sample solution = (TU TS) Ta = (T3 T1) Tb R = vS/vU, the ratio of the high dose of the Standard in USP Units (vS) to the high dose of the Injection in mL (vU) Compute the log confidence interval L (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). Replication: Repeat the entire determination at least once. Test the agreement among the two or more independent determinations, and compute the weight for each (see Design and Analysis of Biological Assays 111, Combination of Independent Assays). Calculate the weighted mean logpotency M and its confidence interval, L (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). The Potency, P, is satisfactory if P = antilog M is 80% 125% of the labeled potency, and if the confidence interval does not exceed 0.40. Acceptance criteria: 80.0%125.0% IMPURITIES Organic Impurities VASOPRESSIN ACTIVITY Solution A: Dissolve 6.6 g of dibasic ammonium phosphate in 950 mL of water, and adjust with concentrated phosphoric acid to a pH of 3.0. Dilute with water to 1 L. Mobile phase: Acetonitrile and Solution A (13:87) [NOTEThe retention time of the vasopressin peak is very sensitive to small changes in the acetonitrile concentration.] Standard solution: Dissolve the entire contents of a vial of USP Vasopressin RS in a known volume of Solution A, and dilute with Solution A to obtain a final solution containing 0.1 USP Vasopressin Units/mL. Sample solution: Dissolve the entire contents of a vial of Injection in a known volume of Solution A, and dilute with Solution A to obtain a final solution containing 2.0 USP Corticotropin Units/mL. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5 m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the vasopressin activity in USP Vasopressin Units/USP Corticotropin Unit: Result = (rU/rS)/2CS rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of the Standard solution (USP Vasopressin Units/mL) i
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NMT 0.05
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containing 8.0 mL of metaphosphoric acid solution (1 in 40), and comminute the glands by grinding with a small quantity of washed sand. Cover each vessel, and proceed similarly until all glands have been extracted. Ascorbic acid determination: Filter the metaphosphoric acid extracts, and pipet 4 mL of each filtrate into suitable vessels each containing 4.0 mL of indophenol-acetate TS. Mix by shaking, and read the absorbance at 520 nm, with a suitable spectrophotometer. From the observed absorbance and the standard curve prepared as directed in the next paragraph, calculate the amount of ascorbic acid in terms of mg of ascorbic acid in each 100 g of adrenal gland tissue. Prepare a standard concentration-absorbance curve, using three Standard solutions containing, in each mL, 6.0, 8.0, and 10.0 g of USP Ascorbic Acid RS in metaphosphoric acid solution (1 in 40), respectively. Pipet into each of three suitable vessels, preferably spectrophotometer cells, 4 mL of indophenol-acetate TS. Add 4.0 mL of one of the three standard ascorbic acid solutions to one of the cells, mix, and promptly read the absorbance in the same instrument and under the same conditions as for the adrenal gland extracts. Repeat the process for the other two standard ascorbic acid solutions, plot the concentration-absorbance values, and draw the straight line best fitting the three plotted points. Calculation: Tabulate the observed concentration of ascorbic acid in the adrenal glands of each rat, designated by the symbol y, for each dosage group of f rats. If the data from one or more rats are missing, adjust to groups of equal size by suitable means (see Design and Analysis of Biological Assays 111, Replacement of Missing Values). Total the values of y in each group, and designate each total as T, subscripts 1 to 3 for the three successive dosage levels and subscripts S and U for the Standard and the Injection, respectively. Test both the agreement in slope of the dosage-response lines for the Standard and for the Injection, and the lack of curvature, as directed for a 3-dose balanced assay (see Design and Analysis of Biological Assays 111, Tests of Assay Validity). If the combined discrepancy as measured by F3 exceeds its tabular value in Table 9 (see Design and Analysis of Biological Assays 111, Combination of Independent Assays), regard these data as preliminary and repeat the Assay. Determine the logarithm of potency of the Injectable Suspension: M = (4iTa/3Tb) + log R = (TU TS) = (T3 T1) = interval between successive log doses of both the Standard solution and the Sample solution R = vS/vU is the ratio of the high dose of the Standard in USP Units (vS) to the high dose of the Injection in mL (vU) Compute the log confidence interval L (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). Replication: Repeat the entire determination at least once. Test the agreement among the two or more independent determinations, and compute the weight for each (see Design and Analysis of Biological Assays 111, Combination of Independent Assays). Calculate the weighted mean logpotency M and its confidence interval, Lc (see Design and Analysis of Biological Assays 111, Confidence Intervals for Individual Assays). The potency, P*, is satisfactory if P* = antilog, M is NLT 80%, and NMT 125% of the labeled potency, and if the confidence interval does not exceed 0.40. Ta Tb i
SPECIFIC TESTS PH 791: 3.07.0 BACTERIAL ENDOTOXINS TEST 85: Contains NMT 3.1 USP Endotoxin Units/USP Corticotropin Unit. OTHER REQUIREMENTS: Meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Ascorbic Acid RS USP Corticotropin RS USP Endotoxin RS USP Vasopressin RS
Corticotropin Zinc Hydroxide Injectable Suspension

(Comment on this Monograph)id=m20190=Corticotropin Zinc Hydroxide Injectable Suspension=Chl-Cy-Monos.pdf) Corticotropin zinc hydroxide [9050-75-3]. DEFINITION Corticotropin Zinc Hydroxide Injectable Suspension is a sterile suspension of corticotropin adsorbed on zinc hydroxide. Its potency is NLT 80.0% and NMT 125.0% of the potency stated on the label in USP Corticotropin Units. It contains NLT 1800 g and NMT 2200 g of zinc, and NLT 604 g and NMT 776 g of anhydrous dibasic sodium phosphate, for each 40 USP Corticotropin Units. ASSAY PROCEDURE Standard solution: Pipet 2.5 mL of gelatin TS into an opened container of USP Corticotropin RS, and mix to obtain a solution having a concentration of 2.0 USP Corticotropin Units/mL. Using gelatin TS as a diluent, prepare three diluted Standard solutions such that the respective concentrations of corticotropin constitute a geometric series such as 1:2:4 or 1:3:9, and such that the quantity of corticotropin in each 0.5 mL lies within the range of 10300 milli-units. Sample solution: Add sufficient 0.1 N hydrochloric acid to Injectable Suspension to effect a complete solution, and prepare in the same manner as the Standard solution, and using the same diluent, dilute the Injectable Suspension to give three Sample solutions corresponding to those of the Standard solution. Animals: Select healthy rats, of the same or either sex, that have been raised on a diet fully adequate with respect to vitamin and mineral content. Anesthetize the rats with ether, and remove the hypophysis from each by application of gentle suction through a fine-tipped tube. Between 16 and 48 h after the operation, select those rats weighing between 80 and 180 g, but restrict the selection so that no rat is more than 30% heavier than the lightest, and the number of rats is an exact multiple of six. Separate the selected rats into six groups, equal in size, of NLT six rats each, and assign at random one of the three diluted Standard solutions or of the three test solutions to each group. Analysis: Inject all rats of each group subcutaneously with the assigned test doses. Anesthetize the rats 3 h after the injection, and remove both adrenal glands from each rat, free them from adhering tissue, and promptly weigh each pair on a suitable balance to the nearest 0.2 mg. Place the weighed glands from each rat in suitable vessels each
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before use. The container label and the package label state the potency in USP Corticotropin Units in each mL. USP REFERENCE STANDARDS 11 USP Ascorbic Acid RS USP Corticotropin RS USP Endotoxin RS
SPECIFIC TESTS ZINC Solution A: 5.4 g of ammonium chloride and 26 mL of ammonium hydroxide. Dilute with water to 100 mL. Sample solution: Transfer the equivalent to 6 mg of zinc from well-shaken Injectable Suspension to a 125-mL conical flask. Add 2 mL of Solution A, 10 mL of water, and 2 drops of eriochrome black TS. Analysis: Titrate with 0.005 M edetate disodium VS to a clear blue endpoint. Perform a blank titration, and make any necessary correction (see Titrimetry 541). Each mL of 0.005 M edetate disodium is equivalent to 0.327 mg of Zn. Acceptance criteria: 18002200 g for 40 USP Corticotropin Units ANHYDROUS DIBASIC SODIUM PHOSPHATE Ammonium molybdate reagent: 64 mg/mL of ammonium molybdate in water and 10 N sulfuric acid (1:1). (Dissolve in water first.) [NOTEThis solution is stable for about 2 weeks.] Stannous chloride reagent: 200 mg/mL of stannous chloride in hydrochloric acid. Just prior to use, dilute 1 mL of the resultant solution with water to 100 mL. Standard solution: 27.5 g/mL of anhydrous dibasic sodium phosphate (Na2HPO4) in water Sample solution: Mix 1 mL of the well-shaken Injectable Suspension with 0.1 mL of 10 N sulfuric acid, and dilute with water to 25 mL. Spectrometric conditions Mode: UV-vis Analytical wavelength: 710 nm Analysis Samples: Standard solution and Sample solution Into separate 100-mL volumetric flasks, pipet duplicate 10mL portions of the Sample solution, duplicate 10-mL portions of the Standard solution, and 10 mL of water to provide a blank. Treat each flask as follows. Dilute the contents with water to 60 mL, then add 10 mL of 10 N sulfuric acid. Add 10 mL of Ammonium molybdate reagent, 10 mL of water, and mix again. Add slowly, with mixing, 5 mL of Stannous chloride reagent, and dilute with water to volume. Determine the absorbances of the resulting solutions of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Measure the absorbances of the solutions at 10 min after the first addition of the Stannous chloride reagent. Calculate the quantity, in g, of anhydrous dibasic sodium phosphate in the portion of Injectable Suspension taken: Weight result (g) = (AU/AS) 25 CS = absorbance of the Sample solution = absorbance of the Standard solution = concentration of Na2HPO4 in the Standard solution (g/mL) Acceptance criteria: 604776 g PH 791: 7.58.5, determined potentiometrically BACTERIAL ENDOTOXINS TEST 85: It contains NMT 3.1 USP Endotoxin Units/USP Corticotropin Unit. OTHER REQUIREMENTS: It meets the requirements under Injections 1. AU AS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. Store at controlled room temperature. LABELING: Label it to indicate that it is not recommended for intravenous use and that the suspension is to be well shaken
Cortisone Acetate
(Comment on this Monograph)id=m20250=Cortisone Acetate=Chl-Cy-Monos.pdf)
C23H30O6 402.48 Pregn-4-ene-3,11,20-trione, 21-(acetyloxy)-17-hydroxy-; 17,21-Dihydroxypregn-4-ene-3,11,20-trione 21-acetate [50-04-4]. DEFINITION Cortisone Acetate contains NLT 97.0% and NMT 102.0% of C23H30O6, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Dissolve in methanol, evaporate the methanol on a steam bath, and dry at 105 for 30 min. B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 238 nm Medium: Methanol Solution: 10 g/mL Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0%. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (45:55) Solution A: Transfer 20 mL of 1 N hydrochloric acid, 150 mL of 0.5 N potassium chloride, and 50 mL of 0.5 M sodium acetate to a 1-L volumetric flask, and dilute with water to volume. Diluent: Acetonitrile and Solution A (1:1) Standard solution: 0.1 mg/mL of USP Coritsone Acetate RS in Diluent [NOTESonicate during preparation to facilitate dissolution.] Sample solution: 0.1 mg/mL of Cortisone Acetate in Diluent [NOTESonicate during preparation to facilitate dissolution.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2.0 Capacity factor k: NLT 2.0 Relative standard deviation: NMT 2.0% for replicate injections
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Acceptance criteria Any individual impurity: NMT 1.5% Total impurities: NMT 2.0%
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the quantity, in percentage, of C23H30O6 in the portion of Cortisone Acetate taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cortisone Acetate RS in the Standard solution (mg/mL) CU = concentration of Cortisone Acetate in the Sample solution (mg/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: Negligible, from 100 mg Organic Impurities PROCEDURE Solution A: Acetonitrile and water (3:7) Solution B: Acetonitrile and water (7:3) Mobile phase: Use variable mixtures of Solution A and Solution B as follows:
rU rS CS
SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +208 to +217 Sample solution: 10 mg/mL, in dioxane LOSS ON DRYING 731: Dry it at 105 for 30 min: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Cortisone Acetate RS
Cortisone Acetate Injectable Suspension

(Comment on this Monograph)id=m20270=Cortisone Acetate Injectable Suspension=Chl-Cy-Monos.pdf) DEFINITION Cortisone Acetate Injectable Suspension is a sterile suspension of Cortisone Acetate in a suitable aqueous medium. It contains NLT 90.0% and NMT 110.0% of the labeled amount of cortisone acetate (C23H30O6). IDENTIFICATION INFRARED ABSORPTION 197K Sample: Mix 25 mL of water with a volume of Injectable Suspension equivalent to 25 mg of cortisone acetate. Centrifuge, or allow the insoluble material to settle, then decant and discard the supernatant. Add 20 mL of methanol and, using agitation and warming as necessary, dissolve the residue. Evaporate the solvent on a steam bath with the aid of a current of air, then dry the residue at 105 for 30 min. ASSAY PROCEDURE Mobile phase: n-Butyl chloride, water-saturated n-butyl chloride, tetrahydrofuran, methanol, and glacial acetic acid (95:95:14:7:6) Internal standard solution: 0.5 mg/mL of prednisone in Mobile phase Standard solution: Transfer 12 mg of USP Cortisone Acetate RS to a stoppered, 50-mL conical flask. Add 20.0 mL of Internal standard solution, and sonicate for 5 min. Pass a portion through a polytef syringe filter, then combine 1 mL of the filtrate and 4 mL of Mobile phase to obtain the Standard solution. System suitability solution: 0.1 mg/mL of hydrocortisone acetate in the Standard solution Sample solution: Transfer 2.0 mL of freshly mixed Injectable Suspension to a volumetric flask of a size to give a nominal cortisone acetate concentration of 2 mg/mL when diluted to volume. Dilute with isopropyl alcohol to volume, and sonicate for 3 min. Deliver a 3.0-mL aliquot of this solution to a stoppered, 25-mL conical flask, and evaporate on a steam bath with the aid of a current of air to dryness. Add 10.0 mL of Internal standard solution, insert the stopper, and sonicate for 5 min. Pass a portion through a polytef syringe filter, then combine approximately 1 mL of the filtrate and 4 mL of Mobile phase to obtain the Sample solution. Chromatographic system (See Chromatography 621, System Suitability.)
Diluent: Filtered mixture of acetonitrile, glacial acetic acid, and water (7:0.1:3) Standard solution: 20 g/mL of USP Cortisone Acetate RS in Diluent Sample solution: 2.5 mg/mL of Cortisone Acetate in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 15 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 5.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of impurity in the portion of Cortisone Acetate taken: Result = (ri/rS) (CS/CU) 100 ri rS CS CU = response for each impurity from the Sample solution = response of the major peak from the Standard solution = concentration of USP Cortisone Acetate RS in the Standard solution (mg/mL) = concentration of Cortisone Acetate in the Sample solution (mg/mL)
USP 32
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L3 Flow rate: 1 mL/min Injection size: 15 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for cortisone acetate and prednisone in the Standard solution are about 0.6 and 1.0, respecitvely.] Suitability requirements Resolution: NLT 2.2 between cortisone acetate and hydrocortisone acetate, System suitability solution [NOTEIf necessary, add equal parts of n-butyl chloride and watersaturated n-butyl chloride to the Mobile phase to meet this requirement.] Relative standard deviation: NMT 2.0% for replicate injections in the Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H30O6 in each mL of the Injectable Suspension taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cortisone acetate to the internal standard from the Sample solution = peak response ratio of cortisone acetate to the RS internal standard from the Standard solution = concentration of USP Cortisone Acetate RS in the CS Standard solution (mg/mL) = nominal concentration of cortisone acetate in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 5.07.0 OTHER REQUIREMENTS: Injections 1. RU
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methanol to the filtrate, mix, evaporate the solvent on a steam bath with the aid of a current of air, then dry the residue at 105 for 30 min. The residue so obtained meets the requirements of the following test. INFRARED ABSORPTION 197K ASSAY PROCEDURE Mobile phase: n-Butyl chloride, water-saturated n-butyl chloride, tetrahydrofuran, methanol, and glacial acetic acid (95:95:14:7:6). Internal standard solution: 0.04 mg/mL of methylparaben in Mobile phase Standard solution: Transfer 12 mg of USP Cortisone Acetate RS to a glass-stoppered, 50-mL conical flask, add 25.0 mL of Internal standard solution, and sonicate for 5 min. Combine about 1 mL of this solution with 3 mL of Mobile phase. System suitability solution: 0.1 mg/mL of hydrocortisone acetate in the Standard solution Sample solution: Weigh, then finely powder, NLT 20 Tablets. Transfer a portion of the powder, nominally equivalent to 12 mg of cortisone acetate, to a stoppered conical flask. Add 25.0 mL of Internal standard solution, insert the stopper in the flask, and sonicate vigorously for 5 min. Pass a portion through a polytef syringe filter, then combine about 1 mL of the filtrate and 3 mL of Mobile phase. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6 mm 25 cm; packing L3 Flow rate: 1 mL/min Injection size: 15 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for methylparaben and cortisone acetate are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2.2 between cortisone acetate and hydrocortisone acetate, System suitability solution [NOTEIf necessary, add equal parts of n-butyl chloride and watersaturated n-butyl chloride to the Mobile phase to meet this requirement.] Relative standard deviation: NMT 2.0% for replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H30O6 in the portion of Tablets taken: Result = (RU/RS) (CS/CU) 100 RU RS CS CU = peak response ratio of cortisone acetate to the internal standard from the Sample solution = peak response ratio of cortisone acetate to the internal standard from the Standard solution = concentration of USP Cortisone Acetate RS in the Standard solution (mg/mL) = nominal concentration of cortisone acetate in the Sample solution (mg/mL)
It meets the requirements under
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Cortisone Acetate RS
Cortisone Acetate Tablets

(Comment on this Monograph)id=m20290=Cortisone Acetate Tablets=Chl-Cy-Monos.pdf) DEFINITION Cortisone Acetate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of cortisone acetate (C23H30O6). IDENTIFICATION [NOTEThe Sample is prepared as follows.] Sample: Powder a number of Tablets, equivalent to 25 mg of cortisone acetate, add 25 mL of solvent hexane, and extract for 15 min with occasional agitation. Decant and discard the supernatant, then extract the residue with 5 mL of chloroform, with frequent agitation, for 5 min. Filter, add 10 mL of
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90.0%110.0%
USP 32
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281 Sample: 5 g of Purified cotton Analysis: Place Sample in a porcelain or platinum dish, and moisten with 2 N sulfuric acid. Gently heat the cotton until it is charred, then ignite more strongly until the carbon is completely consumed. Acceptance criteria: NMT 0.20% of residue remains. Organic Impurities Procedure 1 Fatty Matter Analysis: Pack 10 0.01 g in a Soxhlet extractor provided with a tared receiver, and extract with ether for 5 h at a rate such that the ether siphons over NLT four times per h. The ether solution in the flask shows no trace of blue, green, or brownish color. Evaporate the extract to dryness, and dry at 105 for 1 h. Acceptance criteria: The weight of the residue is NMT 70 mg (0.7%). Procedure 2 Dyes Analysis: Pack 10 g in a narrow percolator, and extract slowly with alcohol until the percolate measures 50 mL. Acceptance criteria: When observed downward through a column 20 cm in depth, the percolate may show a yellowish color but neither a blue nor a green tint. Procedure 3 Other Foreign Matter: The pinches of it taken for the determination of Fiber length contain no oil stains or metallic particles. SPECIFIC TESTS ACIDITY OR ALKALINITY Sample: 10 g of Purified cotton Analysis: Thoroughly saturate Sample with 100 mL of recently boiled and cooled water, then with the aid of a glass rod, press out two 25-mL portions of the water into white porcelain dishes. To one portion add 3 drops of phenolphthalein TS, and to the other portion add 1 drop of methyl orange TS. Acceptance criteria: No pink color develops in either portion. STERILITY TESTS 71: Meets the requirements WATER-SOLUBLE SUBSTANCES Analysis: Place 10 g in a beaker containing 1000 mL of water, and boil gently for 30 min, adding water as required to maintain the volume. Pour the water through a funnel into another vessel, and press out the excess water from the cotton with a glass rod. Wash the cotton in the funnel with two 250-mL portions of boiling water, pressing the cotton after each washing. Filter the combined extract and washings, and wash the filter thoroughly with hot water. Evaporate the combined extract and washings to a small volume, transfer to a tared porcelain or platinum dish, evaporate to dryness, and dry the residue at 105 to constant weight. Acceptance criteria: The residue weighs NMT 35 mg (0.35%). COTTON, Fiber Length 691 and Absorbency Test 691: Remove it from its wrappings, and condition it for NLT 4 h in a standard atmosphere of 65 2% relative humidity at 21 1.1 (70 2F), before determining the Fiber length and Absorbency. Fiber length Analysis: Determine the fiber length of Purified Cotton as directed under Cotton 691, Fiber Length. Acceptance criteria: NLT 60% of the fibers by weight are 12.5 mm or greater in length, and NMT 10% of the fibers by weight are 6.25 mm or less in length.
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.5% of sodium lauryl sulfate solution; 1000 mL Apparatus 2: 50 rpm Time: 45 min Standard solution: 5.55 g/mL of USP Cortisone Acetate RS in Medium Sample solution: Sample per Dissolution 711 . Dilute with Medium as needed. Spectrometric conditions Mode: UV Analytical wavelength: 242 nm Cell size: 1 cm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C23H30O6 is dissolved. UNIFORMITY OF DOSAGE UNITS, Content Uniformity 905 Mobile phase, Internal standard solution, Standard solution, System suitability solution, Chromatographic system, and System suitability: Proceed as directed in the Assay. Sample solution: Place 1 Tablet in a stoppered, 50-mL conical flask, deposit 0.25 mL of water on the tablet, insert the stopper in the flask, and allow to stand for 30 min. Add 2.5 mL of isopropyl alcohol, and place the unstoppered flask on a steam bath. Boil gently, if necessary, until the tablet disintegrates, then evaporate the solvent with the aid of a current of air. Remove from the steam bath, add 10.0 mL of Internal standard solution for each 5 mg of cortisone acetate declared, insert the stopper, and sonicate vigorously for 10 min. Pass a portion through a polytef syringe filter, then combine about 1 mL of the filtrate and 3 mL of Mobile phase. Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H30O6 in the Tablet taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cortisone acetate to the internal standard from the Sample solution = peak response ratio of cortisone acetate to the RS internal standard from the Standard solution = concentration of USP Cortisone Acetate RS in the CS Standard solution (mg/mL) = nominal concentration of cortisone acetate in CU the Sample solution (mg/mL) Acceptance criteria: Meet the requirements in the chapter. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cortisone Acetate RS RU
Purified Cotton
(Comment on this Monograph)id=m20400=Purified Cotton=Chl-Cy-Monos.pdf) DEFINITION Purified Cotton is the hair of the seed of cultivated varieties of Gossypium hirsutum Linn , or of other species of Gossypium e (Fam. Malvaceae), freed from adhering impurities, deprived of fatty matter, bleached, and sterilized in its final container.
USP 32
Absorbency Analysis: Proceed as directed under Cotton 691, Absorbency Test. Acceptance criteria: Submersion is complete in 10 s at a temperature of 25, and the cotton retains NLT 24 times its weight of water. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Package it in rolls of NMT 500 g of a continuous lap, with a light-weight paper running under the entire lap, the paper being of such width that it may be folded over the edges of the lap to a distance of at least 25 mm, the two together being tightly and evenly rolled, and enclosed and sealed in a well-closed container. It may be packaged also in other types of containers if these are so constructed that the sterility of the product is maintained. LABELING: Its label bears a statement to the effect that the sterility cannot be guaranteed if the package bears evidence of damage or if the package has been opened previously.
Official Monographs / Cromolyn 197

solution into a 100-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. Spectrometric conditions Mode: UV Analytical wavelength: 326 nm Cell: 1 cm Blank: A solution (1 in 100) of Solution A Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H14Na2O11 in the Cromolyn Sodium taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) concentration of the Sample solution (g/mL) Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1: Diluent: Stabilizer-free tetrahydrofuran, acetone and water (4:1:6) Standard solution A: 10 mg/mL of USP Cromolyn Sodium RS in Diluent Standard solution B: 0.05 mg/mL of USP Cromolyn Sodium RS from Standard solution A in Diluent Sample solution: 10 mg/mL of Cromolyn Sodium in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography. Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform, methanol, and glacial acetic acid (9:9:2) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed under General Chapter. Locate the spots on the plate by viewing under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A. Any spot from the Sample solution moving ahead of the principal spot is not more intense than the spot from Standard solution B (0.5%). PROCEDURE 2: LIMIT OF OXALATE Dissolve 100 mg in 20 mL of water, add 5.0 mL of iron salicylate TS, and dilute with water to 50 mL. Determine the absorbance of the solution at 480 nm against a water blank. The absorbance is NLT that of a solution containing 350 g of oxalic acid prepared in the same manner (0.35%). SPECIFIC TESTS ACIDITY OR ALKALINITY: Dissolve 1.0 g in 25 mL of carbon dioxide-free water, and add two drops of bromothymol blue TS. If the solution is yellow, NMT 0.25 mL of 0.1 N sodium hydroxide is required to produce a blue color. If the solution is blue, NMT 0.25 mL of 0.1 N hydrochloric acid is required to produce a yellow color. AU AS CS CU = = = =
Cromolyn Sodium
(Comment on this Monograph)id=m20530=Cromolyn Sodium=Chl-Cy-Monos.pdf)
C23H14Na2O11 512.34 4H-1-Benzopyran-2-carboxylic acid, 5,5-[(2-hydroxy-1,3propanediyl)bis(oxy)bis[4-oxo-, disodium salt]; Disodium 5,5-[(2-hydroxytrimethylene)dioxy]bis[4-oxo-4H-1benzopyran-2-carboxylate] [15826-37-6]. DEFINITION Cromolyn Sodium contains NLT 98.0% and NMT 101.0% of C23H14Na2O11, calculated on the anhydrous basis. IDENTIFICATION A. The IR absorption spectrum of a potassium bromide dispersion of it, previously dried in a vacuum at 105 to constant weight, exhibits maxima only at the same wavelengths as that of a similar preparation of USP Cromolyn Sodium RS. B. The UV absorption spectrum of a solution (1 in 40,000) in Solution A prepared as directed in the Assay exhibits maxima at the same wavelengths as that of a similar solution of USP Cromolyn Sodium RS, concomitantly measured. ASSAY PROCEDURE Solution A: 70 g of anhydrous dibasic sodium phosphate in 900 mL of water. Adjust to a pH of 7.4 by the addition of dilute phosphoric acid (1 in 10). Dilute with water to 1000 mL. Transfer 10 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Standard solution: 250 g/mL of USP Cromolyn Sodium RS in water. Transfer 10 mL of the resultant solution to a 100-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. The final concentration is 25 g/mL. Sample solution: Transfer 100 mg of Cromolyn Sodium to a 1000-mL volumetric flask, dissolve in 100 mL of water, and dilute with water to volume. Pipet 25 mL of this
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NMT 10.0% USP REFERENCE STANDARDS 11 USP Cromolyn Sodium RS
USP 32
WATER DETERMINATION, Method I 921:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cromolyn Sodium RS
Cromolyn Sodium Inhalation Solution

(Comment on this Monograph)id=m20539=Cromolyn Sodium Inhalation Solution=Chl-Cy-Monos.pdf) DEFINITION Cromolyn Sodium Inhalation Solution is a sterile, aqueous solution of Cromolyn Sodium. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C23H14Na2O11. IDENTIFICATION The UV absorption spectrum of the Sample solution prepared as directed in the Assay exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Cromolyn Sodium RS, concomitantly measured. ASSAY PROCEDURE Solution A: 70 g of anhydrous dibasic sodium phosphate in 900 mL of water. Adjust to a pH of 7.4 by adding dilute phosphoric acid (1 in 10). Dilute with water to 1000 mL. Transfer 10 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Standard solution: 250 g/mL of USP Cromolyn Sodium RS. Transfer 10 mL of the resultant solution to a 100-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. The final concentration is 25 g/mL. Sample solution: Equivalent to 250 g/mL of Cromolyn Sodium from Inhalation Solution. Pipet 10 mL of resultant solution to a 100-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. Spectrometric conditions Mode: UV Analytical wavelength: 326 nm Cell: 1 cm Blank: Solution A (1 in 100) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H14Na2O11 in the aliquot taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of cromolyn sodium in the Sample solution (g /mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements AU AS CS CU = = = =
Cromolyn Sodium Inhalation Powder

(Comment on this Monograph)id=m20540=Cromolyn Sodium Inhalation Powder=Chl-Cy-Monos.pdf) DEFINITION Cromolyn Sodium Inhalation Powder is a mixture of equal parts of Lactose and Cromolyn Sodium contained in a hard gelatin capsule. It contains NLT 95.0% and NMT 125.0% of the labeled amount of cromolyn sodium (C23H14Na2O11). IDENTIFICATION ULTRAVIOLET ABSORPTION: The UV absorption spectrum of a solution (1 in 40,000) in Solution A prepared as directed in the Assay exhibits maxima at the same wavelengths as that of a similar solution of USP Cromolyn Sodium RS, concomitantly measured. ASSAY PROCEDURE Solution A: 70 g of anhydrous dibasic sodium phosphate in 900 mL of water. Adjust to a pH of 7.4 by adding dilute phosphoric acid (1 in 10). Dilute with water to 1000 mL. Transfer 10 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Standard solution: 350 g/mL of USP Cromolyn Sodium RS (previously dried in a vacuum at 105 to constant weight). Transfer 10 ml of this solution to a 100-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. The final concentration is 35 g/mL. Sample solution: Remove and weigh the contents of NLT 20 capsules of Cromolyn Sodium Inhalation Powder, and transfer the combined contents to a 250-mL volumetric flask. Dissolve in 100 mL of water, and dilute with water to volume. Transfer an aliquot of this solution, nominally equivalent to 8 mg of cromolyn sodium, to a 250-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. Spectrometric conditions Mode: UV Analytical wavelength: 326 nm Cell: 1 cm Blank: Solution A (1 in 250) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H14Na2O11 in the aliquot taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of cromolyn sodium in the Sample solution (g/mL) Acceptance criteria: 95.0%125.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements AU AS CS CU = = = =
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. Avoid excessive heat.
IMPURITIES Organic Impurities PROCEDURE Diluent: Stabilizer-free tetrahydrofuran, acetone, and water (4:1:6) Standard solution A: 10 mg/mL of USP Cromolyn Sodium RS in Diluent Standard solution B: 0.1 mg/mL of USP Cromolyn Sodium RS from Standard solution A in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture
USP 32
Sample solution: 10 mg/mL of cromolyn sodium from Inhalation Solution in Diluent Application volume: 10 L Developing solvent system: Chloroform, methanol, and glacial acetic acid (9:9:2) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Locate the spots on the plate by viewing under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A. Any spot in the chromatogram of the Sample solution moving ahead of the principal spot is not more intense than the spot in the chromatogram of Standard solution B (1.0%). SPECIFIC TESTS PH 791: 4.07.0 STERILITY TESTS 71:
Official Monographs / Cromolyn 199

Transfer an aliquot of this solution nominally equivalent to 8 mg of cromolyn sodium to a 250-mL volumetric flask. Add 2.5 mL of Solution A, and dilute with water to volume. Spectrometric conditions Mode: UV Analytical wavelength: 326 nm Cell: 1 cm Blank: A solution (1 in 100) of Solution A Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H14Na2O11 in the aliquot taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% AU AS CS CU IMPURITIES Organic Impurities PROCEDURE Diluent: Stabilizer-free tetrahydrofuran, acetone, and water (4:1:6) Standard solution A: 10 mg/mL of USP Cromolyn Sodium RS in Diluent Standard solution B: 0.1 mg/mL of USP Cromolyn Sodium RS from Standard solution A in Diluent Sample solution: 10 mg/mL of cromolyn sodium from Nasal Solution in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform, methanol, and glacial acetic acid (9:9:2) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Locate the spots on the plate by viewing under shortwavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A. Any spot in the chromatogram of the Sample solution moving ahead of the principal spot is not more intense than the spot in the chromatogram of Standard solution B (1.0%). SPECIFIC TESTS PH 791: 4.07.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Cromolyn Sodium RS = = = =
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-unit doubleended glass ampuls or in low-density polyethylene ampuls. LABELING: The label indicates that the Inhalation Solution is not to be used if it contains a precipitate. USP REFERENCE STANDARDS 11 USP Cromolyn Sodium RS
Cromolyn Sodium Nasal Solution

(Comment on this Monograph)id=m20544=Cromolyn Sodium Nasal Solution=Chl-Cy-Monos.pdf) DEFINITION Cromolyn Sodium Nasal Solution is an aqueous solution of Cromolyn Sodium. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C23H14Na2O11. It may contain suitable stabilizers. IDENTIFICATION PROCEDURE The UV absorption spectrum of a solution (1 in 40,000) in Solution A prepared as directed in the Assay exhibits maxima at the same wavelengths as that of a similar solution of USP Cromolyn Sodium RS, concomitantly measured. ASSAY PROCEDURE Solution A: 70 g of anhydrous dibasic sodium phosphate in 900 mL of water. Adjust to a pH of 7.4 by the addition of dilute phosphoric acid (1 in 10). Dilute with water to 1000 mL. Transfer 10 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Standard solution: 250 g/mL of USP Cromolyn Sodium RS in water. Transfer 10 mL of the resultant solution to a 100-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. The final concentration is about 25 g/mL. Sample solution: Transfer 4 mL of Nasal Solution to a 100mL volumetric flask, and dilute with water to volume.
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USP 32
Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform, methanol, and glacial acetic acid (9:9:2) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow the spots to dry, and develop the chromatogram in the Developing solvent system until the solvent has moved three-fourths of the length of the plate. Locate the spots on the plate by viewing under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A. Any spot in the chromatogram of the Sample solution moving ahead of the principal spot is not more intense than the spot in the chromatogram of Standard solution B (1.0%). SPECIFIC TESTS PH 791: 4.07.0 STERILITY TESTS 71:
Cromolyn Sodium Ophthalmic Solution

(Comment on this Monograph)id=m20546=Cromolyn Sodium Ophthalmic Solution=Chl-Cy-Monos.pdf) DEFINITION Cromolyn Sodium Ophthalmic Solution is a sterile, aqueous solution of Cromolyn Sodium. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C23H14Na2O11. It may contain suitable antimicrobial and stabilizing agents. IDENTIFICATION The UV absorption spectrum of a solution (1 in 40,000) in Solution A prepared as directed in the Assay exhibits maxima at the same wavelengths as that of a similar solution of USP Cromolyn Sodium RS, concomitantly measured. ASSAY PROCEDURE Solution A: Dissolve 70 g of anhydrous dibasic sodium phosphate in 900 mL of water. Adjust to a pH of 7.4 by the addition of dilute phosphoric acid (1 in 10). Dilute with water to 1000 mL, and mix. Transfer 10 mL of this solution to a 100-mL volumetric flask, and dilute with water to volume. Standard solution: Dissolve a weighed quantity of USP Cromolyn Sodium RS in water to obtain a solution having a concentration of 250 g/mL. Transfer 10 mL of this solution to a 100-mL volumetric flask, add 1 mL of Solution A, and dilute with water to volume. The final concentration is 25 g/mL. Sample solution: Transfer 4 mL of Ophthalmic Solution to a 100-mL volumetric flask, dilute with water to volume, and mix. Transfer an aliquot of this solution, nominally equivalent to 8 mg of cromolyn sodium, to a 250-mL volumetric flask. Add 2.5 mL of Solution A, and dilute with water to volume. Spectrometric conditions Mode: UV Analytical wavelength: 326 nm Cell: 1 cm Blank: A solution (1 in 100) of Solution A Analysis Samples: Standard solution and Sample solution Calculate the percentage of C23H14Na2O11 in each mL of the Ophthalmic Solution taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Cromolyn Sodium RS in the Standard solution (g/mL) = concentration of cromolyn sodium in the Sample CU solution (g/mL) Acceptance criteria: 90.0%110.0% AU AS CS IMPURITIES Organic Impurities PROCEDURE Diluent: Stabilizer-free tetrahydrofuran, acetone, and water (4:1:6) Standard solution A: 10 mg/mL of USP Cromolyn Sodium RS in Diluent Standard solution B: 0.1 mg/mL of USP Cromolyn Sodium RS in Diluent Sample solution: 10 mg/mL of cromolyn sodium equivalent Ophthalmic Solution in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant, single-dose or multiple-dose containers. Ophthalmic Solution that is packaged in multiple-dose containers contains a suitable antimicrobial agent. USP REFERENCE STANDARDS 11 USP Cromolyn Sodium RS
Crotamiton
(Comment on this Monograph)id=m20640=Crotamiton=ChlCy-Monos.pdf)
C13H17NO 2-Butenamide, N-ethyl-N-(2-methylphenyl)-; N-Ethyl-o-crotonotoluidide [483-63-6].
203.28
DEFINITION Crotamiton is a mixture of cis and trans isomers containing NLT 97.0% and NMT 103.0% of C13H17NO. IDENTIFICATION A. INFRARED ABSORPTION 197F B. ULTRAVIOLET ABSORPTION 197U Sample solution: 20 g/mL in cyclohexane C. PROCEDURE To 10 mL of a saturated solution in water add a few drops of potassium permanganate TS: a brown color is produced, and a brown precipitate is formed on standing. ASSAY PROCEDURE Standard solution: cyclohexane
20 g/mL of USP Crotamiton RS in
USP 32
Sample solution: 20 g/mL of Crotamiton in cyclohexane Spectrometric conditions Mode: UV Analytical wavelength: 242 nm Cell: 1 cm Blank: Cyclohexane Analysis Samples: Standard solution and Sample solution Calculate the percentage of C13H17NO in the portion taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of the Standard solution (g/mL) = concentration of the Sample solution (g/mL) CU Acceptance criteria: 97.0%103.0% AU AS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
Official Monographs / Cupric 201

paper. Pipet 10 mL of the clear filtrate and 5 mL of Internal standard solution into a 50-mL volumetric flask, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm stainless steel column; packing L1 System suitability Sample: Standard solution Suitability requirements Resolution: NLT 3.0 between crotamiton and butyl benzoate Lowest and highest peak response ratios (RS): NMT 2.0% from three replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C13H17NO in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of crotamiton to the butyl benzoate peak from the Sample solution = peak response ratio of crotamiton to the butyl RS benzoate peak from the Standard solution = concentration of USP Crotamiton RS in the CS Standard solution (mg/mL) = nominal concentration of crotamiton in the CU Sample solution (mg/mL) Acceptance criteria: 93.0%107.0% SPECIFIC TESTS MINIMUM FILL 755: Meets the requirements RU
NMT 0.1%
SPECIFIC TESTS SPECIFIC GRAVITY 841: 1.0081.011 at 20 REFRACTIVE INDEX 831: 1.5401.543 at 20 BOUND HALOGEN: Place 4 drops in a 3-mm (ID) test tube, and add calcium oxide to a height of 1 cm. Heat the tube in a flame, starting from the top, until the reaction is complete, then ignite for a short time. Transfer the contents to a beaker containing 10 mL of water, acidify with nitric acid, and filter. To the filtrate add 0.2 mL of silver nitrate solution (1 in 60): any opalescence obtained is NMT that obtained from a blank solution treated in the same manner. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Crotamiton RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Crotamiton RS
Crotamiton Cream
(Comment on this Monograph)id=m20650=Crotamiton Cream=Chl-Cy-Monos.pdf) DEFINITION Crotamiton Cream contains NLT 93.0% and NMT 107.0% of the labeled amount of C13H17NO. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Sample solution. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:2). Internal standard solution: 17.5 mg/mL of butyl benzoate in methanol Standard stock solution: 1 mg/mL of USP Crotamiton RS in methanol Standard solution: Pipet 10 mL of Standard stock solution and 5 mL of Internal standard solution into a 50-mL volumetric flask, and dilute with methanol to volume. Sample solution: Transfer a portion of Crotamiton Cream, nominally equivalent to 50 mg of crotamiton, to a tared 50mL volumetric flask. Add 25 mL of methanol, and shake and sonicate to disperse the cream. Dilute with methanol to volume. Pass 20 mL through moderately retentive filter
Cupric Chloride
(Comment on this Monograph)id=m20685=Cupric Chloride=Chl-Cy-Monos.pdf) CuCl2 2H2O Copper chloride (CuCl2) dihydrate; Copper(2+) chloride dihydrate [10125-13-0]. Anhydrous [7447-39-4]. 170.48 134.45
DEFINITION Cupric Chloride contains NLT 99.0% and NMT 100.5% of CuCl2, calculated on the dried basis. IDENTIFICATION IDENTIFICATION TESTSGENERAL, Chloride 191 and Copper 191: A solution (1 in 20) meets the requirements for the tests. ASSAY PROCEDURE Sample solution: 400 mg of Cupric Chloride in 50 mL of water. Add 4 mL of acetic acid and 3 g of potassium iodide. Analysis: Titrate the liberated iodine with 0.1 N sodium thiosulfate VS, adding 2 g of potassium thiocyanate and 3 mL of starch TS as the endpoint is approached. Each mL of 0.1 N sodium thiosulfate is equivalent to 13.45 mg of CuCl2.
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99.0%100.5%
Limit Test Iron Nickel Sample Weight (g) 2.0 2.0
USP 32
IMPURITIES Inorganic Impurities LIMIT OF SODIUM Stock solution: Transfer 10.0 g of Cupric Chloride to a 100-mL volumetric flask, add water, and swirl to dissolve. Add 5 mL of nitric acid, dilute with water to volume, and mix. Sample solutions: To three 25-mL volumetric flasks, add a volume of Stock solution equivalent to the weight of the Sample in the table below. To two of the flasks add the amounts of reference analyte ion specified in the table below. Add 2 mL of 50 mg/mL potassium chloride solution to each flask, and dilute with water to volume. Analysis: Using atomic absorption spectrophotometry (see Spectrophotometry and Light-Scattering 851), analyze the Sample solutions by the method of standard addition analysis according to the table below. Acceptance criteria: NMT 0.02% LIMIT OF IRON Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: To three 25-mL volumetric flasks, add a volume of Stock solution equivalent to the weight of Sample in the table below. To two of the flasks, add the amounts of reference analyte ion specified in the table below, and dilute with water to volume. Analysis: Use the Analysis from the test for Limit of sodium. Acceptance criteria: NMT 0.005% LIMIT OF NICKEL Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: Use the Sample solutions from the test for the Limit of iron. Analysis: Use the Analysis from the test for Limit of sodium. Acceptance criteria: NMT 0.01% SULFATE Analysis: Heat to boiling the combined filtrate and washings retained from the test for Insoluble matter, add 10 mL of barium chloride TS, digest for 2 h on a steam bath, and allow to stand overnight. Filter the solution through a tared, medium-porosity porcelain filtering crucible, and wash the residue with two 10-mL portions of hot water. Ignite at 800 25 to constant weight. Acceptance criteria: The weight of the residue, corrected for the weight obtained in a blank test, does not exceed 1.2 mg (0.005%). LIMIT OF POTASSIUM Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: Use the Sample solutions from the test for the Limit of iron. Analysis: Use the Analysis from the test for Limit of sodium. Acceptance criteria: NMT 0.01% LIMIT OF CALCIUM Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: Use the Sample solutions from the test for the Limit of iron. Analysis: Use the Analysis from the test for Limit of sodium. Acceptance criteria: NMT 0.005%
Wavelength (nm) 589.0 766.5 422.7 Sample Weight (g) 0.1 0.1 2.0 Reference Ion Added (mg) 0.01/0.02 0.01/0.02 0.05/0.10
Wavelength (nm) 248.3 232.0
Reference Ion Added (mg) 0.05/0.10 0.10/0.20
correction is No except for iron; for iron, it is Yes.] INSOLUBLE MATTER Sample: 10 g Analysis: Transfer Sample to a 250-mL beaker, add 100 mL of water and 2 mL of hydrochloric acid, cover the beaker, and heat to boiling. Digest the hot solution on a steam bath for 1 h, and filter through a tared, fine-porosity filtering crucible. Rinse the beaker with hot water, passing the rinsings through the filter, and finally wash the filter with additional hot water. [NOTERetain the combined filtrate and washings for the test for Sulfate.] Dry the filter at 105. Acceptance criteria: Residue weighs NMT 1.0 mg (0.01%). SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 105 for 16 h: it loses between 20.9% and 21.4% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25; excursions are permitted between 15 and 30.
[NOTEThe flame type is air-acetyline; the background
Cupric Chloride Injection

(Comment on this Monograph)id=m20690=Cupric Chloride Injection=Chl-Cy-Monos.pdf) DEFINITION Cupric Chloride Injection is a sterile solution of Cupric Chloride in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of copper (Cu). IDENTIFICATION The Sample solution exhibits an absorption maximum at about 325 nm when prepared and tested as directed in the Assay. ASSAY PROCEDURE Sodium chloride solution: 1.35 g/L of sodium chloride Standard stock solution: Tranfser 1.000 g of copper to a 1000-mL volumetric flask, dissolve in 20 mL of nitric acid, and dilute with 0.2 N nitric acid to volume. This solution contains 1000 g of copper per mL. Store in a polyethylene bottle. Standard solutions: Pipet 15 mL of Standard stock solution into a 250-mL volumetric flask, dilute with water to volume, and mix. Transfer 4.0, 5.0, and 6.0 mL of this solution to separate 100-mL volumetric flasks containing 10 mL of Sodium chloride solution, dilute the contents of each flask with water to volume, and mix. These Standard solutions contain 2.4, 3.0, and 3.6 g of copper/mL respectively. Sample stock solution: Transfer a volume of Injection, equivalent to 2 mg of copper from Injection, in 100 mL of water. Sample solution: Pipet 15 mL of the Sample stock solution into a 100-mL volumetric flask. From the labeled amount of sodium chloride, if any, in the Injection, calculate the amount, in mg, of sodium chloride in the initial dilution, and add sufficient Sodium chloride solution to bring the total sodium content of this flask to 13.5 mg. Dilute with water to volume. Spectrophotometric conditions: (See Spectrophotometry and Light-scattering 851.)
Limit Test Sodium Potassium Calcium
USP 32
Mode: Atomic adsorption Lamp: copper hollowcathode Flame: Airacetylene Analytical wavelength: 324.8 nm (copper emission line) Blank: Sodium chloride solution and water (1:9) Analysis: Samples: Standard solutions and Sample solution Plot the absorbances of the Standard solutions versus concentration, in g/mL, of copper, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, in g/mL, of copper in the Sample solution. Calculate the percentage of copper in the Injection: Result = ([C/V] 100 F V1 D)/L = concentration of copper in the Sample solution (g/mL) V = volume of Injection (mL) F = conversion factor from g to mg, 1/1000 = volume of the Sample stock solution, 100 mL V1 D = dilution factor from the Sample solution, 100/15 mL L = label claim (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 1.52.5 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 250.0 USP Endotoxin Units/mg of copper. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I or Type II glass. LABELING: Label the Injection to indicate that it is to be diluted to the appropriate strength with Sterile Water for Injection or other suitable fluid prior to administration. USP REFERENCE STANDARDS 11 USP Endotoxin RS C
Official Monographs / Cupric 203

allow to cool in a desiccator, and weigh again to obtain the weight of the sample. Dissolve in 50 mL of water, and add 4 mL of 6 N acetic acid and 3 g of potassium iodide. Analysis: Titrate the liberated iodine with 0.1 N sodium thiosulfate VS, adding about 2 g of potassium thiocyanate and 3 mL of starch TS as the endpoint is approached. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N sodium thiosulfate is equivalent to 15.96 mg of CuSO4. Acceptance criteria: 98.5%100.5% IMPURITIES Inorganic Impurities LIMIT OF SODIUM Stock solution: Transfer 40.0 g of Cupric Sulfate to a 200mL volumetric flask, add water, and swirl to dissolve. Add 5 mL of nitric acid, and dilute with water to volume. This solution contains 0.2 g of cupric sulfate per mL. Sample solutions: To three 25-mL volumetric flasks, add a volume of Stock solution equivalent to the weight of the Sample in the table below. To two of the flasks, add the amounts of reference analyte ion specified in the table. Add 2 mL of 50 mg/mL of potassium chloride solution to each flask, and dilute with water to volume. Analysis: Using atomic absorption spectrophotometry (see Spectrophotometry and Light-Scattering 851), analyze the Sample solutions by the method of standard addition analysis according to the table below. Acceptance criterioa: NMT 0.02% LIMIT OF CALCIUM Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: To three 25-mL volumetric flask, add a volume of Stock solution equivalent to the weight of the Sample in the table below. To two of the flasks, add the amounts of reference analyte ion specified in the table, and dilute with water to volume. Analysis: Use the Analysis under the test for Limit of sodium. Acceptance criteria: NMT 0.005% LIMIT OF IRON Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: Use the Sample solutions prepared for the test for Limit of calcium. Analysis: Use the Analysis under the test for Limit of sodium. Acceptance criteria: NMT 0.003% LIMIT OF NICKEL Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: Use the Sample solutions prepared for the test for Limit of calcium. Procedure: Use the Analysis under the test for Limit of sodium. Acceptance criteria: NMT 0.005% LIMIT OF POTASSIUM Stock solution: Use the Stock solution prepared in the test for Limit of sodium. Sample solutions: Use the Sample solutions prepared for the test for Limit of calcium. Procedure: Use the Analysis under the test for Limit of sodium.
Cupric Sulfate
(Comment on this Monograph)id=m20700=Cupric Sulfate=ChlCy-Monos.pdf) CuSO4 5H2O Sulfuric acid, copper(2+) salt (1:1), pentahydrate; Copper(2+) sulfate (1:1) pentahydrate [7758-99-8] Anhydrous [7758-98-7]. 249.69 159.61
DEFINITION Cupric Sulfate, dried at 250 to constant weight, contains NLT 98.5% and NMT 100.5 % of CuSO4. IDENTIFICATION A. IDENTIFICATION TESTSGENERAL, Sulfate 191: (1 in 10) meets the requirements. B. IDENTIFICATION TESTSGENERAL, Copper 191: (1 in 10) meets the requirements. A solution A solution
ASSAY PROCEDURE Sample solution: Place 650 mg of Cupric Sulfate in a weighed container fitted with a ground-glass stopper, dry,
204
Cupric / Official Monographs

NMT 0.01%
Sample Weight (g) 0.05 0.4 0.8 4.0 4.0 Reference Ion Added (mg) 0.005/0.01 0.02/0.04 0.05/0.10 0.12/0.24 0.10/0.20
USP 32
Mode: Atomic adsorption Lamp: Copperhollow cathode Flame: Airacetylene Analytical wavelength: 324.8 nm (copper emission line) Blank: 1 in 10 dilution of Sodium chloride solution Analysis Samples: Standard solutions and Sample solution Plot the absorbances of the Standard solutions versus concentration, in g/mL, of copper, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration, in g/mL, of copper in the Sample solution. Calculate the percentage of Cu in the Injection taken: Result = (C/DV) F 100/L = concentration of copper in the Sample solution (g/mL) D = dilution factor for the Injection (0.0015) V = volume of Injection taken to prepare the Sample solution (mL) F = conversion from g to mg (1/1000) L = label claim in mg/mL of Cu in the Injection Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 2.03.5 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 250.0 USP Endotoxin Units/mg of copper PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I or Type II glass. LABELING: Label the Injection to indicate that it is to be diluted to the appropriate strength with Sterile Water for Injection or other suitable fluid prior to administration. USP REFERENCE STANDARDS 11 USP Endotoxin RS C
Limit Test Sodium Potassium Calcium Iron Nickel
Wavelength (nm) 589.0 766.5 422.7 248.3 232.0
[NOTE The flame type is air-acetylene and the
background correction is No except for Iron, which is Yes.]
SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 250 to constant weight: it loses between 33.0% and 36.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at 25; excursions are permitted between 15 and 30.
Cupric Sulfate Injection

(Comment on this Monograph)id=m20703=Cupric Sulfate Injection=Chl-Cy-Monos.pdf) DEFINITION Cupric Sulfate Injection is a sterile solution of Cupric Sulfate in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of Cu. IDENTIFICATION The Sample solution exhibits an absorption maximum at about 325 nm when prepared and tested as directed for Analysis in the Assay. ASSAY PROCEDURE Sodium chloride solution: 1.35 mg/mL of sodium chloride in water Copper stock solution: 1 g of copper in 20 mL of nitric acid, dilute with 0.2 N nitric acid to 1000 mL [NOTEThis solution contains 1000 g/mL of copper. Store in a polyethylene bottle.] Standard solutions: Dilute 50 mL of Copper stock solution to 250 mL. Transfer 4.0, 5.0, and 6.0 mL of the resultant solution to separate 100-mL volumetric flasks containing 10 mL of Sodium chloride solution, dilute the contents of each flask with water to volume, and mix. These Standard solutions contain, respectively, 2.4, 3.0, and 3.6 g/mL of copper. Sample solution: Transfer an amount of Injection nominally equivalent to 2 mg of copper to a 100-mL volumetric flask and dilute with water to volume. Pipet 15 mL of the resultant solution into a 100-mL volumetric flask. From the labeled amount of sodium chloride, if any, in the Injection, calculate the amount, in mg, of sodium chloride in the initial dilution, and add sufficient Sodium chloride solution to bring the total sodium content of this flask to 13.5 mg. Dilute with water to volume. Spectrometric conditions (See Spectrometry and Light Scattering 851.)
Cyanocobalamin
(Comment on this Monograph)id=m20750=Cyanocobalamin=Chl-Cy-Monos.pdf)
C63H88CoN14O14P Vitamin B12 [68-19-9].
1355.37
DEFINITION Cyanocobalamin contains NLT 96.0% and NMT 100.5% of C63H88CoN14O14P, calculated on the dried basis. IDENTIFICATION A. The absorption spectrum of the solution employed for measurement of absorbance in the Assay exhibits maxima
USP 32
within 1 nm at 278 nm and 361 nm and within 2 nm at 550 nm. The ratio A361/A278 is between 1.70 and 1.90, and the ratio A361/A550 is between 3.15 and 3.40. B. PROCEDURE Sample: 1 mg cyanocobalamin Analysis: Fuse the sample with 50 mg of potassium pyrosulfate in a porcelain crucible. Cool, break up the mass with a glass rod, add 3 mL of water, and dissolve by boiling. Add 1 drop of phenolphthalein TS, and add 100 mg/mL of sodium hydroxide solution, dropwise, until just pink. Add 500 mg of sodium acetate, 0.5 mL of 1 N acetic acid, and 0.5 mL of 2 mg/mL nitroso R salt solution. Acceptance criteria: A red or orange-red color appears at once. Add 0.5 mL of hydrochloric acid, and boil for 1 min: the red color persists. C. PROCEDURE Sample: 5 mg of cyanocobalamin Analysis: Dissolve Sample in 5 mL of water in a 50-mL distilling flask connected with a short, water-cooled condenser. To the flask add 2.5 mL of hypophosphorous acid, close, boil gently but short of distillation for 10 min, then distill 1 mL into a test tube containing 1 mL of 20 mg/mL sodium hydroxide solution. To the test tube add 4 drops of cold saturated ferrous ammonium sulfate solution, shake gently, then add 30 mg of sodium fluoride, and bring the contents to a boil. Immediately add, dropwise, 5 N sulfuric acid until a clear solution results, then add 35 drops more of the acid. Acceptance criteria: A blue or blue-green color develops within a few min. ASSAY PROCEDURE Standard solution: 30 g/mL of USP Cyanocobalamin RS Sample solution: 30 g/mL of Cyanocobalamin Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 361 nm Cell: 1 cm Blank: Water Analysis Samples: Standard solution and Sample solution Determine the absorbances of the Standard solution and the Sample solution at the wavelength of maximum absorbance. Calculate the percentage of C63H88CoN14O14P in the portion of Cyanocobalamin taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Cyanocobalamin RS in the Standard solution (g/mL) CU = concentration of Cyanocobalamin in the Sample solution (g/mL) Acceptance criteria: 96.0%100.5% AU AS CS IMPURITIES Organic Impurities PROCEDURE: PSEUDO CYANOBOBALAMIN Sample: 1.0 mg of Cyanocobalamin Analysis: Dissolve Sample in 20 mL of water contained in a small separator, add 5 mL of a mixture of carbon tetrachloride and m-cresol (1:1), and shake well for about 1 min. Allow to separate, draw off the lower layer into a second small separator, add 5 mL of 5 N sulfuric acid, shake well, and allow to separate completely. [NOTEThe complete separation of the layer may be facilitated by centrifuging.]
Official Monographs / Cyanocobalamin 205

Acceptance criteria: The separated upper layer is colorless, or has no more color than a mixture of 0.15 mL of 0.10 N potassium permanganate and 250 mL of water. SPECIFIC TESTS LOSS ON DRYING 731: Heat 25 mg in a suitable vacuum drying apparatus at 105, and at a pressure of NMT 5 mm of mercury for 2 h. Cool, and weigh: it loses NMT 12.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cyanocobalamin RS
Cyanocobalamin Injection
(Comment on this Monograph)id=m20760=Cyanocobalamin Injection=Chl-Cy-Monos.pdf) DEFINITION Cyanocobalamin Injection is a sterile solution of Cyanocobalamin in Water for Injection, or in Water for Injection rendered isotonic by the addition of Sodium Chloride. It contains NLT 95.0% and NMT 115.0% of the labeled amount of anhydrous cyanocobalamin (C63H88CoN14O14P). IDENTIFICATION PROCEDURE Samples: Standard solution and Sample solution from the Assay Spectrometric conditions Mode: UV-Vis Analytical wavelength: 300550 nm Blank: 1 in 10 dilution of Sodium chloride solution Acceptance criteria: The Sample solution exhibits maxima at the same wavelengths as that of the Standard solution, concomitantly measured, and the ratio A361/A550 = 3.153.40. ASSAY PROCEDURE Standard solution: 30 g/mL of USP Cyanocobalamin RS Sample solution: Nominally equivalent to 30 g/mL of cyanocobalamin from Injection (dilute a volume of Injection of NLT 300 g of cyanocobalamin) in water Spectrometric conditions Mode: UV Analytical wavelength: 361 nm Cell: 1 cm Blank: Water Analysis: Calculate the percentage of C63H88CoN14O14P in the portion of Injection taken: Result = (AU/AS) (CS/CU) 100 absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) nominal concentration of anhydrous cyanocobalamin in the Sample solution (g/mL) Acceptance criteria: 95.0%115.0% AU AS CS CU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.4 USP Endotoxin Unit/g of cyanocobalamin. = = = =
206
Cyanocobalamin / Official Monographs
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Relative standard deviation: NMT 2.0% for replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H24O3 in the portion of Cyclandelate taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cyclandelate RS in the Standard solution (mg/mL) CU = concentration of Cyclandelate in the Sample solution (mg/mL) Acceptance criteria: NLT 98.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Mobile phase: Acetonitrile and water (4:1) Standard solution: Pipet 3.0 mL of the Sample solution into a 100-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution: 1 mg/mL of Cyclandelate in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 228 nm Column: 4.0-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 7.0 between cyclandelate and dicyclohexyl phthalate, System suitability solution Relative standard deviation: NMT 2.0% for replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Allow the chromatogram of the Sample solution to run for a period of time that is about 3 times the retention time of cyclandelate. Acceptance criteria: The total area of all the peaks from the Sample solution, other than the peak of cyclandelate, is NMT the peak area of cyclandelate of the Standard solution. (NMT 3.0% of total impurities is found.) SPECIFIC TESTS LOSS ON DRYING 731 Sample: 1 g Analysis: Dry the Sample over silica gel for 24 h. Acceptance criteria: The Sample loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store below 40, preferably between 15 and 30. USP REFERENCE STANDARDS 11 USP Cyclandelate RS
PH 791: 4.57.0 OTHER REQUIREMENTS: Injections 1.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in light-resistant, singledose or multiple-dose containers, preferably of Type I glass, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cyanocobalamin RS USP Endotoxin RS
Cyclandelate
(Comment on this Monograph)id=m20890=Cyclandelate=ChlCy-Monos.pdf)
C17H24O3 276.38 3,3,5-Trimethylcyclohexanol -phenyl--hydroxyacetate; 1,5-cis-3,3,5-Trimethylcyclohexyl 2-hydroxy-2-phenyl acetate [456-59-7]. DEFINITION Cyclandelate contains NLT 98.0% of C17H24O3, calculated on the dried basis. IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Sample solution: 0.5 mg/mL Medium: 95% alcohol Acceptance criteria: The solution exhibits absorption maxima between 250 and 254 nm, between 256 and 260 nm, and between 262 and 266 nm. B. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 10 mg of Cyclandelate in 1 mL of alcohol Application volume: 5 L Developing solvent system: Hexane, ethyl acetate, and glacial acetic acid (8:2:1) ASSAY PROCEDURE Mobile phase: Acetonitrile and water (4:1) System suitability solution: 0.2 mg/mL of USP Cyclandelate RS and 0.08 mg/mL of dicyclohexyl phthalate in Mobile phase Standard solution: 0.2 mg/mL of USP Cyclandelate RS in Mobile phase Sample solution: 0.2 mg/mL of Cyclandelate in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 228 nm Column: 4.0-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 7.0 between cyclandelate and dicyclohexyl phthalate, System suitability solution
USP 32
Official Monographs / Cyclizine 207

IDENTIFICATION A. PROCEDURE Sample: Amount equivalent to 500 mg of cyclizine hydrochloride, from finely powdered Tablets Analysis: Shake the Sample with 25 mL of water for 5 min, and filter the mixture. Cool the filtrate in an ice bath, add a slight excess of 1N sodium hydroxide, and stir well. Wash the precipitate of the base so obtained with water, and dry in a vacuum at 60 for 2 h. Acceptance criteria: The precipitate of the base so obtained melts between 106 and 109. B. IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Meet the requirements ASSAY SALTS OF ORGANIC NITROGENOUS BASES 501 Analysis Samples: Standard solution and Sample solution Proceed with Tablets as directed, diluting the Standard solution and the Sample solution, respectively, with an equal volume of dilute sulfuric acid (1 in 100), and determining the absorbance at the wavelength of maximum absorbance at 264 nm. Calculate the percentage of C18H22N2 HCl taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Cyclizine Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of cyclizine hydrochloride CU in Sample solution (mg/mL) Acceptance criteria: NLT 93.0% and NMT 107.0% AU AS CS PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 900 mL Apparatus 2: 50 rpm Time: 45 min Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Analysis: Determine the amounts of C18H22N2 HCl dissolved by employing the Analysis set forth in the Assay, making any necessary volumetric adjustments. Tolerances: NLT 75% (Q) of the labeled amount of C18H22N2 HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Cyclizine Hydrochloride RS
Cyclizine Hydrochloride
(Comment on this Monograph)id=m20950=Cyclizine Hydrochloride=Chl-Cy-Monos.pdf) C18H22N2 HCl 302.84 Piperazine, 1-(diphenylmethyl)-4-methyl-, monohydrochloride; 1-(Diphenylmethyl)-4-methylpiperazine monohydrochloride [303-25-3]. DEFINITION Cyclizine Hydrochloride contains NLT 98.0% and NMT 100.5% of C18H22N2 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample: 500 mg Analysis: Dissolve the Sample in 10 mL of a mixture of 3 volumes of alcohol and 2 volumes of water, warming if necessary. Cool the solution in an ice bath, add 1 mL of 1 N sodium hydroxide and 20 mL of water, stir, and filter. Wash the precipitate of the base so obtained with water, and dry in a vacuum at 60 for 2 h. Acceptance criteria: The precipitate of the base so obtained melts between 106 and 109. C. IDENTIFICATION TESTSGENERAL, Chloride 191 Acceptance criteria: Meets the requirements ASSAY PROCEDURE Sample: 400 mg Analysis: Transfer the Sample to a beaker, and dissolve it in 80 mL of glacial acetic acid. Add 10 mL of mercuric acetate TS, and titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 15.14 mg of C18H22N2 HCl. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Sample solution: Methanol Standard solution: Methanol Eluant: Chloroform, methanol, and ammonium hydroxide (80:20:1) Visualization: 2 SPECIFIC TESTS PH 791 Sample solution: 1 in 50 in a mixture of alcohol and water (2:3) as solvent Acceptance criteria: 4.55.5 LOSS ON DRYING 731: Dry a sample at 120 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Cyclizine Hydrochloride RS
Cyclizine Hydrochloride Tablets

(Comment on this Monograph)id=m20980=Cyclizine Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Cyclizine Hydrochloride Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of C18H22N2 HCl.
208
Cyclobenzaprine / Official Monographs
USP 32
solvent system until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, air-dry, and view under shortwavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of Standard solution A; and any other spot of the Sample solution does not exceed, in size or intensity, the principal spot of Standard solution B (0.5%). SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 215219, but the range between beginning and end of melting does not exceed 2. LOSS ON DRYING 731: Dry it at 105 to constant weight: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cyclobenzaprine Hydrochloride RS
Cyclobenzaprine Hydrochloride
(Comment on this Monograph)id=m21060=Cyclobenzaprine Hydrochloride=Chl-Cy-Monos.pdf)
C20H21N HCl 311.85 1-Propanamine, 3-(5H-dibenzo[a,d]cyclohepten-5-ylidene)-N,Ndimethyl-, hydrochloride; N,N-Dimethyl-5H-dibenzo[a,d]cycloheptene-5,-propylamine hydrochloride [6202-23-9]. DEFINITION Cyclobenzaprine Hydrochloride contains NLT 99.0% and NMT 101.0% of C20H21N HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: At 290 nm Medium: Methanol Solution: 15 g/mL Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 3.0%. C. IDENTIFICATION TESTSGENERAL, Chloride 191: A solution (1 in 50) meets the requirements. ASSAY PROCEDURE Sample solution: 400 mg of Cyclobenzaprine Hydrochloride in 80 mL of glacial acetic acid. Add 15 mL of mercuric acetate TS. Analysis: Titrate with 0.1 N perchloric acid VS, using a platinum ring electrode and a sleeve-type calomel electrode containing 0.1 N lithium perchlorate in glacial acetic acid (see Titrimetry 541). Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 31.19 mg of C20H21N HCl. Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 0.001% Organic Impurities PROCEDURE Standard solution A: 20 mg/mL of USP Cyclobenzaprine Hydrochloride RS in methanol Standard solution B: 100 g/mL of USP Cyclobenzaprine Hydrochloride RS from Standard solution A in methanol Sample solution: 20 mg/mL in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture [NOTEWash the thin-layer chromatographic plate with methanol before use.] Application volume: 5 L Developing solvent system: Acetone, toluene, and ammonium hydroxide (75:25:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Develop the chromatogram in a suitable chamber with a freshly prepared Developing
Cyclobenzaprine Hydrochloride Tablets

(Comment on this Monograph)id=m21090=Cyclobenzaprine Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Cyclobenzaprine Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of cyclobenzaprine hydrochloride (C20H21N HCl). IDENTIFICATION A. INFRARED ABSORPTION 197M Sample: Transfer an equivalent to 50 mg of cyclobenzaprine hydrochloride, from a quantity of finely powdered Tablets, to a small flask, add 10 mL of methylene chloride, swirl to dissolve, and filter. Evaporate the clear filtrate to about 5 mL, transfer to a suitable centrifuge tube, and add 12 mL of ether. Evaporate with the aid of a current of air to about 1 mL, and agitate until crystallization occurs. Wash the crystals with several portions of ether, and air-dry. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, methanesulfonic acid, and water (28:24:0.2:48); adjust with diethylamine to a pH of 3.6. Standard solution: 0.05 mg/mL of USP Cyclobenzaprine Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: Prepare a quantity equivalent to 0.05 mg/mL of cyclobenzaprine hydrochloride from finely powdered Tablets (NLT 20) in 0.1 N hydrochloric acid. [NOTEShake by mechanical means for 30 min.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 290 nm Column: 4.6-mm 10-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: NLT 2.0 for the analyte peak Column efficiency: NLT 1000 theoretical plates from the analyte peak
USP 32
Tailing factor: NMT 2 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C20H21N HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 peak response from the Sample solution peak response from the Standard solution concentration of the Standard solution (mg/mL) nominal concentration of cyclobenzaprine hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS CU PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 1: 50 rpm Time: 30 min Standard solution: USP Cyclobenzaprine Hydrochloride RS in Medium Sample solution: Sample per Dissolution 711. Filter and dilute with Medium as needed. Spectrometric conditions Mode: UV Analytical wavelength: 290 nm Analysis Samples: Standard solution and Sample Solution Tolerances: NLT 75% (Q) of the labeled amount of C20H21N HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cyclobenzaprine Hydrochloride RS = = = =
Official Monographs / Cyclopentolate 209

Mobile phase: Acetonitrile and Solution A (7:3) Standard solution: 0.1 mg/mL of USP Cyclopentolate Hydrochloride RS Sample solution: 0.1 mg/mL of Cyclopentolate Hydrochloride Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L15 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3000 theoretical plates from the analyte peak Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C17H25NO3 HCl in the portion of Cyclopentolate Hydrochloride taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cyclopentolate Hydrochloride RS in the Standard solution (mg/mL) CU = concentration of Cyclopentolate Hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.05% Organic Impurities PROCEDURE Solution A, Mobile phase, and Chromatographic system: Prepare as directed in the Assay. Sample solution: Use the Sample solution prepared as directed in Assay. Analysis Sample: Sample solution Record the chromatogram obtained for a period of NLT twice the retention time of cyclopentolate, and measure the peak responses. Calculate the percentage of each peak, other than the solvent peak and the cyclopentolate peak, in the portion of Cyclopentolate Hydrochloride by the same formula: Result = (ri/rT) 100 = response of each peak = sum of the responses of all of the peaks, excluding that of the solvent peak Acceptance criteria Individual impurity: NMT 1.0% Total impurities: NMT 2.0% ri rT SPECIFIC TESTS PH 791: 4.55.5, in a solution (1 in 100) LOSS ON DRYING 731: Dry at 105 for 4 h: it loses NMT 0.5% of its weight. rU rS CS
Cyclopentolate Hydrochloride
(Comment on this Monograph)id=m21290=Cyclopentolate Hydrochloride=Chl-Cy-Monos.pdf)
327.85 C17H25NO3 HCl Benzeneacetic acid, -(1-hydroxycyclopentyl)-, 2(dimethylamino)ethyl ester, hydrochloride, ()-; 2-(Dimethylamino)ethyl ()-1-hydroxy-phenylcyclopentaneacetate hydrochloride [5870-29-1]. DEFINITION Cyclopentolate Hydrochloride contains NLT 98.0% and NMT 102.0% of C17H25NO3 HCl, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. IDENTIFICATION TESTSGENERAL, Chloride 191: solution (1 in 500)
ASSAY PROCEDURE Solution A: 0.66 mg/mL of dibasic ammonium phosphate in water. Adjust with phosphoric acid to a pH of 3.0 0.1.
210
Cyclopentolate / Official Monographs

CU
USP 32
= nominal concentration of cyclopentolate hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 3.05.5 STERILITY TESTS 71:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store in a cold place. USP REFERENCE STANDARDS 11 USP Cyclopentolate Hydrochloride RS
Cyclopentolate Hydrochloride Ophthalmic Solution

(Comment on this Monograph)id=m21300=Cyclopentolate Hydrochloride Ophthalmic Solution=Chl-Cy-Monos.pdf) DEFINITION Cyclopentolate Hydrochloride Ophthalmic Solution is a sterile, aqueous solution of Cyclopentolate Hydrochloride. It may contain suitable buffers and other additives. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C17H25NO3 HCl. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Place in a 125-mL separator a volume of Ophthalmic Solution, equivalent to 50 mg of cyclopentolate hydrochloride. Add 1 g of potassium carbonate, and extract with two 10-mL portions of ether. Pass the ether extracts through ether-washed filter paper, collect the filtrate in a small beaker, and evaporate to dryness. Standard: Place in a second separator 50 mg of USP Cyclopentolate Hydrochloride RS dissolved in 5 mL of water. Follow the procedure under Sample. ASSAY PROCEDURE Solution A: 0.66 mg/mL of dibasic ammonium phosphate in water. Adjust with phosphoric acid to a pH of 3.0 0.1. Mobile phase: Acetonitrile and Solution A (7:3) Standard solution: 0.1 mg/mL of USP Cyclopentolate Hydrochloride RS Sample solution: 0.1 mg/mL of cyclopentolate hydrochloride from Ophthalmic Solution diluted with water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 15-cm; packing L15 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3000 theoretical plates Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage label claim of C17H25NO3 HCl in the Ophthalmic Solution taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = cyclopentolate peak response from the Sample solution = cyclopentolate peak response from the Standard solution = concentration of USP Cyclopentolate Hydrochloride RS in the Standard solution (mg/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cyclopentolate Hydrochloride RS
Cyclophosphamide
(Comment on this Monograph)id=m21350=Cyclophosphamide=Chl-CyMonos.pdf)
279.10 C7H15Cl2N2O2P H2O 261.09 C7H15Cl2N2O2P 2H-1,3,2-Oxazaphosphorin-2-amine, N,N-bis(2chloroethyl)tetrahydro-, 2-oxide, monohydrate, (); ()-2-[Bis(2-chloroethyl)amino]tetrahydro-2H-1,3,2oxazaphosphorine 2-oxide monohydrate [6055-19-2]. Anhydrous [50-18-0]. DEFINITION Cyclophosphamide contains NLT 97.0% and NMT 103.0% of C7H15Cl2N2O2P, calculated on the anhydrous basis. [CAUTIONGreat care should be taken in handling Cyclophosphamide, as it is a potent cytotoxic agent.] IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:7) Internal standard solution: Dissolve 185 mg of ethylparaben in 250 mL of alcohol in a 1000-mL volumetric flask, and dilute with water to volume. Standard solution: Transfer USP Cyclophosphamide RS, equivalent to 25 mg of anhydrous cyclophosphamide, to a 50-mL volumetric flask, add 25 mL of water, and shake to dissolve the USP Reference Standard. Add 5.0 mL of Internal standard solution, dilute with water to volume, and mix to obtain a Standard solution having a concentration of 0.5 mg/mL of anhydrous cyclophosphamide. Sample solution: Nominally equivalent to 1 mg/mL of anhydrous cyclophosphamide from Cyclophosphamide. Shake for 5 min. Pipet 25 mL of this resultant solution and 5 mL of Internal standard solution into a 50-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 195 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for cyclophosphamide and ethylparaben are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2 between cyclophosphamide and ethylparaben Relative standard deviation: NMT 2% from six replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H15Cl2N2O2P in the Cyclophosphamide taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cyclophosphamide to the ethylparaben peak in the Sample solution = peak response ratio of cyclophosphamide to the RS ethylparaben peak in the Standard solution = concentration of the Standard solution (mg/mL) CS = concentration of Cyclophosphamide in the CU Sample solution (mg/mL) Acceptance criteria: 97.0%103.0% IMPURITIES Inorganic Impurities HEAVY METALS 231: NMT 20 ppm Sample solution: 40 mg/mL, and filter if necessary SPECIFIC TESTS PH 791: 3.97.1, in a solution (1 in 100), determined 30 min after its preparation WATER DETERMINATION, Method I 921: 5.7%6.8% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers at a temperature between 2 and 30. USP REFERENCE STANDARDS 11 USP Cyclophosphamide RS RU
Official Monographs / Cyclophosphamide 211

Acceptance criteria: Meets the requirements B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, both relative to the internal standard, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:7) Internal standard solution: Dissolve 185 mg of ethylparaben in 250 mL of alcohol in a 1000-mL volumetric flask, and dilute with water to volume. Standard solution: Transfer USP Cyclophosphamide RS, equivalent to 25 mg of anhydrous cyclophosphamide, to a 50-mL volumetric flask, add 25 mL of water, and shake to dissolve the USP Reference Standard. Add 5.0 mL of Internal standard solution, dilute with water to volume, and mix to obtain a Standard solution having a concentration of 0.5 mg/mL of anhydrous cyclophosphamide. Sample solution: Nominally equivalent to 1 mg/mL of anhydrous cyclophosphamide from Injection diluted with water. Shake for 5 min. Pipet 25 mL of this resultant solution and 5 mL of Internal standard solution into a 50-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 195 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for cyclophosphamide and ethylparaben are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2 between cyclophosphamide and ethylparaben Relative standard deviation: NMT 2% from six replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H15Cl2N2O2P in the portion of Cyclophosphamide for Injection taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of cyclophosphamide to the ethylparaben peak in the Sample solution RS = peak response ratio of cyclophosphamide to the ethylparaben peak in the Standard solution = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements RU
Cyclophosphamide for Injection

(Comment on this Monograph)id=m21360=Cyclophosphamide for Injection=Chl-Cy-Monos.pdf) DEFINITION Cyclophosphamide for Injection is a sterile mixture of Cyclophosphamide with or without a suitable diluent. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C7H15Cl2N2O2P. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: 20 mg/mL of cyclophosphamide in chloroform Standard solution: 20 mg/mL USP Cyclophosphamide in chloroform Application volume: 5 L Developing solvent system: Chloroform, methanol, and ammonium hydroxide (15:4:1) Analysis Samples: Standard solution and Sample solution Visualize the spots by placing the plate in an iodine chamber.
SPECIFIC TESTS PH 791: 3.09.0, but the range does not exceed 3 pH units, in a solution containing the equivalent of 20 mg of anhydrous cyclophosphamide per mL, determined 30 min after its preparation. BACTERIAL ENDOTOXINS TEST 85: NMT 0.20 USP Endotoxin Unit/mg of cyclophosphamide INJECTIONS, Constituted Solutions 1: At the time of use, it meets the requirements.
212
Cyclophosphamide / Official Monographs
USP 32
Mode: LC Detector: UV 195 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for cyclophosphamide and ethylparaben are 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 2 between cyclophosphamide and ethylparaben Relative standard deviation: NMT 2% from six replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C7H15Cl2N2O2P per Tablet taken: Result = (RU/RS) (CS/CU) 100 = ratio of the peak response of cyclophosphamide to that of ethylparaben in the Sample solution = ratio of the peak response of cyclophosphamide RS to that of ethylparaben in the Standard solution CS = concentration of the Standard solution corrected for moisture by a titrimetric water determination (mg/mL) = nominal concentration of cyclophosphamide in CU the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL, deaerated Apparatus 1: 100 rpm Time: 45 min Determine the amount of C7H15Cl2N2O2P dissolved. Mobile phase: Acetonitrile and water (3:7) Standard solution: USP Cyclophosphamide RS in water Sample solution: Sample per Dissolution 711. Dilute with Medium as needed. Pass through a filter having a porosity of 0.8 m. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 195 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the amount of C7H15Cl2N2O2P dissolved: Result = (rU/rS) CS V (100/L) rU = cyclophosphamide peak response from the Sample solution
STERILITY TESTS 71: Meets the requirements INJECTIONS, Labeling 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve as described in Containers for Sterile Solids under Injections 1. Storage at a temperature not exceeding 25 is recommended. It will withstand brief exposure to temperatures up to 30 but is to be protected from temperatures above 30. USP REFERENCE STANDARDS 11 USP Cyclophosphamide RS USP Endotoxin RS
Cyclophosphamide Tablets
(Comment on this Monograph)id=m21370=Cyclophosphamide Tablets=Chl-Cy-Monos.pdf) DEFINITION Cyclophosphamide Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of anhydrous cyclophosphamide (C7H15Cl2N2O2P). IDENTIFICATION A. INFRARED ABSORPTION Sample: Extract a portion of finely powdered Tablets, equivalent to 50 mg of cyclophosphamide, with 25 mL of chloroform; filter about 2 mL of the chloroform solution; mix the filtrate with 500 mg of potassium bromide; evaporate the chloroform, carefully removing the last trace of solvent in a small vacuum flask; and use the residue to prepare a potassium bromide dispersion. Acceptance criteria: The IR absorption spectrum of the Sample exhibits maxima, between 6.5 and 14 m, only at the same wavelengths as that of a similar preparation of USP Cyclophosphamide RS. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:7) Internal standard solution: Dissolve 185 mg of ethylparaben in 250 mL of alcohol in a 1000-mL volumetric flask, and dilute with water to volume. Standard solution: Transfer a quantity of USP Cyclophosphamide RS, equivalent to 25 mg of anhydrous cyclophosphamide, to a 50-mL volumetric flask, add 25 mL of water, and shake to dissolve. Add 5.0 mL of Internal standard solution, dilute with water to volume, and mix to obtain a Standard solution having a known concentration of 0.5 mg/mL of anhydrous cyclophosphamide. Sample solution: Transfer NLT 10 Tablets to a volumetric flask of suitable size so that the final concentration is nominally 1 mg/mL of anhydrous cyclophosphamide. Fill about half full with water, shake for 30 min, and dilute with water to volume. Pass through fast, fluted filter paper, discarding the first 4050 mL of the filtrate. Pipet 25 mL of the filtrate and 5 mL of Internal standard solution into a 50mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
= cyclophosphamide peak response from the Standard solution = concentration of the Standard solution (mg/mL) CS V = volume of Medium, 900 L = tablet label claim (mg) Tolerances: NLT 75% (Q) of the labeled amount of C7H15Cl2N2O2P is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ANALYSIS FOR CONTENT UNIFORMITY Perchloric acid solution: Perchloric acid and water (23.5:976.5) 4-(p-Nitrobenzyl)pyridine solution: 7.5 mg/mL of 4-(pnitrobenzyl)pyridine in ethylene glycol Sodium hydroxide solution: 20 mg/mL of sodium hydroxide in diluted alcohol Standard solution: 500 g/mL of USP Cyclophosphamide RS Sample solution: Place 1 Tablet in a volumetric flask of suitable size so that the final concentration is 500 g/mL. Fill the flask about two-thirds full of water, shake until the Tablet is completely disintegrated, dilute with water to volume, and filter, discarding the first 10 mL of the filtrate. Spectrometric conditions Mode: UV-Vis Analytical wavelength: 560 nm Analysis Samples: Standard solution and Sample solution Place 2.0 mL each of the Sample solution and Standard solution in separate 27-mm 170-mm test tubes, with 2.0 mL of water in each tube as the blank. Treat each tube as follows. Add 0.7 mL of Perchloric acid solution, mix, and heat at 95 for 10 min. Cool, add 1.0 mL of sodium acetate TS, mix, add 1.6 mL of 4-(p-Nitrobenzyl)pyridine solution, mix, and heat at 95 for 10 min. Cool, and add 8.0 mL of Sodium hydroxide solution. Within 4 min after preparation, determine the absorbances of the solutions. Calculate the percentage of C7H15Cl2N2O2P in the Tablet taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = = = = absorbance of the Sample solution absorbance of the Standard solution concentration of the Standard solution (g/mL) concentration of the Sample solution (g/mL) rS
Official Monographs / Cyclopropane 213

DEFINITION Cyclopropane contains NLT 99.0%, by volume, of C3H6. [CAUTIONCyclopropane is highly flammable. Do not use where it may be ignited.] ASSAY PROCEDURE Sample: Cyclopropane Analysis: Place the container so that when its valve is opened, the gaseous phase can be sampled. Withdraw 100 mL of Cyclopropane, measured in a gas buret previously filled with mercury and equipped with a leveling bulb at the lower end. Connect one arm of the buret stopcock to a pipet that previously has been filled with sulfuric acid. By appropriate manipulation of the stopcock and the leveling bulb, transfer the gas between the pipet and the buret, bringing about sufficient contact of the gas with the acid to reduce the volume of unabsorbed gas to a minimum as measured in the buret. NMT 1.0 mL of gas remains. Acceptance criteria: NLT 99.0%, by volume IMPURITIES Inorganic Impurities HALOGENS Sample: Cyclopropane Analysis: Provide a 500-mL flask with a tightly fitting twohole stopper. Through one opening, pass a delivery tube bent at right angles and extending just beyond the lower surface of the stopper. Through the other opening, insert a capillary tube bent at right angles and having a bore of 1 0.2 mm, in the same manner. Place in a 50-mL cylinder, having an internal diameter of 2 0.25 cm, 40 mL of a solution containing 850 mg of sodium carbonate in 1000 mL of water. Provide the cylinder with a two-hole stopper. Through one opening pass a right-angle delivery tube, having a bore of 3 0.5 mm, to within 2 mm of the bottom of the cylinder. The end of the delivery tube that extends out of the cylinder is provided with an enlargement 8 0.5 cm long, having an internal diameter of 2 0.25 cm. Through the other opening in the stopper, pass another right-angle delivery tube, having it extend just below the surface of the stopper. Collect 500 mL of Cyclopropane in the flask. By means of hydrostatic pressure, applied through the delivery tube, force the gas through the capillary tube, the water used being previously saturated with Cyclopropane. Ignite the gas, place the enlarged end of the delivery tube, connected with the cylinder, around the flame, extending the flame one-third of the way into the enlargement. Apply suction to the shorter delivery tube connected with the cylinder, thus drawing the spent gases through the sodium carbonate solution: the period of ignition of the 500 mL of Cyclopropane requires about 30 min. Make any necessary correction for the amount of halogen in the volume of air used for the ignition of the gas. Transfer the sodium carbonate solution to a 500-mL volumetric flask, and rinse the cylinder thoroughly, collecting the rinsings in the flask. Dilute the solution to volume. To a 50-mL aliquot add sufficient nitric acid to make it acid to litmus paper, and then add 1 mL of acid in excess. Prepare a blank containing 0.50 mL of 0.0012 N hydrochloric acid and 4 mL of the sodium carbonate solution in 46 mL of water, acidify to litmus with nitric acid, then add 1 mL of acid in excess and 1 mL of silver nitrate TS to each solution. Acceptance criteria: After 5 min any opalescence in the solution representing the Cyclopropane does not exceed that in the blank (0.02% as chloride).
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Storage at a temperature not exceeding 25 is recommended. Tablets will withstand brief exposure to temperatures up to 30, but are to be protected from temperatures above 30. USP REFERENCE STANDARDS 11 USP Cyclophosphamide RS
Cyclopropane
(Comment on this Monograph)id=m21420=Cyclopropane=ChlCy-Monos.pdf)
C3H6 Cyclopropane [75-19-4].
42.08
214
Cyclopropane / Official Monographs
USP 32
DEFINITION Cycloserine has a potency of NLT 900 g of C3H6N2O2 per mg. IDENTIFICATION PROCEDURE Sample solution: 0.1 mg/mL in 0.1 N sodium hydroxide Analysis: To 1 mL of the Sample solution, add 3 mL of 1 N acetic acid and 1 mL of a mixture that is prepared 1 h before use and is comprised of equal parts of 40 mg/mL sodium nitroprusside solution and 4 N sodium hydroxide. Acceptance criteria: A blue color gradually develops. ASSAY PROCEDURE pH 6.8 phosphate buffer: Prepare as directed in Reagents, Indicators, and SolutionsBuffer Solutions. Mobile phase: Dissolve 0.5 g of sodium 1-decanesulfonate in 800 mL of water. Add 50 mL of acetonitrile and 5 mL of glacial acetic acid, and mix. Adjust with 1 N sodium hydroxide to a pH of 4.4. Standard solution: 0.4 mg/mL of USP Cycloserine RS in pH 6.8 phosphate buffer Sample solution: 0.4 mg/mL of Cycloserine in pH 6.8 phosphate buffer Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 219 nm Column: 4.6-mm 25-cm; 5-m packing L1 Column temperature: 30 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.8 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C3H6N2O2 in each mg taken: Result = (rU/rS) (CS/CU) F = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cycloserine RS in the Standard solution (mg/mL) = concentration of Cycloserine in the Sample CU solution (mg/mL) F = unit conversion factor, 1000 g/g Acceptance criteria: NLT 900 g/mg IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.5%, the charred residue being moistened with 2 mL of nitric acid and 5 drops of sulfuric acid SPECIFIC TESTS CONDENSATION PRODUCTS: Its absorptivity (see Spectrophotometry and Light-Scattering 851) at 285 nm, determined in a 0.1 N sodium hydroxide solution containing 0.40 mg/mL, is NMT 0.80. rU rS CS
Organic Impurities PROCEDURE 1: CARBON DIOXIDE Sample: Cyclopropane Analysis: Place the container so that when its valve is opened, the gaseous phase can be sampled. Connect one end of a carbon dioxide detector tube to the container valve, and the other end to a gas flow meter. Pass 1000 mL of the Cyclopropane through the tube at a suitable rate. Acceptance criteria: The indicator change corresponds to NMT 0.03%. PROCEDURE 2: PROPYLENE, ALLENE, AND OTHER UNSATURATED HYDROCARBONS Sample: Cyclopropane Analysis: Place the container so that when its valve is opened, the gaseous phase can be sampled. Connect one end of an olefin detector tube to the container valve, and the other end to a gas flow meter. Pass the Cyclopropane through the detector tube at a suitable rate. Acceptance criteria: The color of the indicating layer of the tube contents matches the color standard after the passage of NLT 400 mL of Cyclopropane (0.9% as propylene). SPECIFIC TESTS ACIDITY OR ALKALINITY Sample: Cyclopropane Analysis: Add 0.3 mL of methyl red TS and 0.3 mL of bromothymol blue TS to 400 mL of boiling water, and boil the solution for 5 min. Pour 100 mL of the boiling solution into each of three color-comparison tubes marked A, B, and C, respectively. To tube B add 0.20 mL of 0.012 N hydrochloric acid, and to tube C add 0.40 mL of 0.012 N hydrochloric acid. Insert the stopper in each of the tubes, and cool them to room temperature. Pass 2000 mL of Cyclopropane through the solution in tube B at a rate requiring about 30 min for the passage of the gas. Acceptance criteria: The color of the solution in tube B is no deeper orange-red than that of the solution in tube C and no deeper yellow-green than that of the solution in tube A. [NOTEThe various detector tubes called for in the respective tests are listed in Reagents, Indicators, and SolutionsReagents.] ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in cylinders. [NOTE Maintain cylinders of Cyclopropane at 25 2 for NLT 6 h prior to withdrawing samples for the tests and Assay, and correct the results to 25 and 760 mm of mercury.] LABELING: The label bears a warning that cyclopropane is highly flammable and is not to be used where it may be ignited.
Cycloserine
(Comment on this Monograph)id=m21450=Cycloserine=ChlCy-Monos.pdf)
C3H6N2O2 3-Isoxazolidinone, 4-amino-, (R)-; (+)-4-Amino-3-isoxazolidinone [68-41-7].
102.09
USP 32
SPECIFIC ROTATION 781S: 108114 Sample solution: 50 mg/mL in 2 N sodium hydroxide CRYSTALLINITY 695: Meets the requirements PH 791: 5.56.5, in a solution (1 in 10) LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in a vacuum at 60 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cycloserine RS CU
Official Monographs / Cyclosporine 215

= nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% PERFORMANCE TESTS DISSOLUTION 711: Determine the amount of C3H6N2O2 dissolved by using the following method. pH 6.8 phosphate buffer, Mobile phase, Chromatographic system, and System suitability: Proceed as directed in the Assay. Medium: pH 6.8 phosphate buffer; 900 mL (see Reagents, Indicators, and SolutionsBuffer Solutions.) Apparatus 1: 100 rpm Time: 30 min Standard solution: 0.25 mg/mL of USP Cycloserine RS in pH 6.8 phosphate buffer Sample solution: Sample per Dissolution 711, at a known concentration Analysis Samples: Standard solution and Sample solution Calculate the percentage of C3H6N2O2 dissolved: Result = (rU/rS) CS V (100/L) = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cycloserine RS in the Standard solution (mg/mL) V = volume of Medium (mL), 900 L = capsule label claim (mg) Tolerances: NLT 80% (Q) of the labeled amount of C3H6N2O2 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS LOSS ON DRYING 731: Dry 100 mg of the contents of Capsules in a capillary-stoppered bottle in a vacuum at 60 for 3 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cycloserine RS rU rS CS
Cycloserine Capsules
(Comment on this Monograph)id=m21460=Cycloserine Capsules=Chl-Cy-Monos.pdf) DEFINITION Cycloserine Capsules contain NLT 90.0% and NMT 120.0% of the labeled amount of cycloserine (C3H6N2O2). IDENTIFICATION PROCEDURE Sample solution: Equivalent to 0.1 mg/mL of cycloserine, from contents of Capsules, in 0.1 N sodium hydroxide, and filter Analysis: To 1 mL of the filtrate, add 3 mL of 1 N acetic acid and 1 mL of a mixture, prepared 1 h before use, of equal parts of 40 mg/mL sodium nitroprusside solution and 4 N sodium hydroxide. Acceptance criteria: A blue color gradually develops. ASSAY PROCEDURE pH 6.8 phosphate buffer: Prepare as directed in Reagents, Indicators, and SolutionsBuffer Solutions. Mobile phase: Dissolve 0.5 g of sodium 1-decanesulfonate in 800 mL of water. Add 50 mL of acetonitrile and 5 mL of glacial acetic acid, and mix. Adjust with 1 N sodium hydroxide to a pH of 4.4. Standard solution: 0.4 mg/mL of USP Cycloserine RS in pH 6.8 phosphate buffer Sample solution: Nominally 0.4 mg/mL filtered solution of cycloserine, from the contents of NLT 20 Capsules, in pH 6.8 phosphate buffer Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 219 nm Column: 4.6-mm 25-cm; 5-m packing L1 Column temperature: 30 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.8 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C3H6N2O2 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 rU rS CS = peak response from the Sample solution = peak response from the Standard solution = concentration of the Standard solution (mg/mL)
Cyclosporine
(Comment on this Monograph)id=m21480=Cyclosporine=ChlCy-Monos.pdf)
1202.61 C62H111N11O12 Cyclo[[(E)-(2S,3R,4R)-3-hydroxy-4-methyl-2-(methylamino)-6octenoyl]-L-2-aminobutyryl-N-methylglycyl-N-methyl-L-leucylL-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-Nmethyl-L-leucyl-N-methyl-L-valyl]; [R-[R*,R*-(E)]]-Cyclic(L-alanyl-D-alanyl-N-methyl-L-leucyl-Nmethyl-L-leucyl-N-methyl-L-valyl-3-hydroxy-N,4-dimethyl-L-2amino-6-octenoyl-L--aminobutyryl-N-methylglycyl-N-methylL-leucyl-L-valyl-N-methyl-L-leucyl) [59865-13-3]. DEFINITION Cyclosporine contains NLT 98.5% and NMT 101.5% of cyclosporine A (C62H111N11O12), calculated on the dried basis.
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Cyclosporine / Official Monographs

Acceptance criteria Any individual impurity: NMT 0.7% Total impurities: NMT 1.5%
USP 32
IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, tert-butyl methyl ether, phosphoric acid, and water (430:50:1:520) Diluent: Acetonitrile and water (1:1) System suitability solution: 1.25 mg/mL of USP Cyclosporine Resolution Mixture RS in Diluent Standard solution A: 1.25 mg/mL of USP Cyclosporine RS in Diluent Standard solution B: 0.01 mg/mL of USP Cyclosporine RS from Standard solution A in Diluent Sample solution: 1.25 mg/mL of cyclosporine in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4-mm 25-cm; 35 m packing L1; connected with 0.25-mm 1-m stainless steel tube Column temperature: 80 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Samples: System suitability solution, Standard solution A, and Standard solution B Suitability requirements Resolution: The cyclosporine U peak and the main cyclosporine peak are resolved from each other. Relative standard deviation: NMT 1.0% from Standard solution A, and NMT 10.0% from Standard solution B Analysis Samples: Standard solution A and Sample solution Calculate the percentage of C62H111N11O12 in the portion taken: Result = (rU/rS) (CS/CU) 100 = main cyclosporine peak area from the Sample solution rS = main cyclosporine peak area from the Standard solution A CS = concentration of Standard solution A (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 98.5%101.5% rU IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Analysis: Use the chromatograms obtained from Standard solution B and the Sample solution in the Assay. Samples: Standard solution B and Sample solution Calculate the percentage of each impurity: Result = (ri/rS) (CS/CU) 100 = peak area of an individual impurity from the Sample solution = main cyclosporine peak area from the Standard rS solution B = concentration of Standard solution B (mg/mL) CS = concentration of the Sample solution (mg/mL) CU [NOTEDiscard any impurities corresponding to less than 0.05%.] ri
SPECIFIC TESTS LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Cyclosporine RS USP Cyclosporine Resolution Mixture RS
Cyclosporine Capsules
(Comment on this Monograph)id=m21481=Cyclosporine Capsules=Chl-Cy-Monos.pdf) DEFINITION Cyclosporine Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of cyclosporine (C62H111N11O12). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE FOR LIQUID CAPSULES Mobile phase: Acetonitrile, methanol, water, and phosphoric acid (550:50:400:0.5) Standard solution: 1 mg/mL of USP Cyclosporine RS in dehydrated alcohol [NOTEUse this solution promptly after preparation.] Sample solution: Using a sharp blade, carefully cut open NLT 20 Capsules. With the aid of dehydrated alcohol, transfer the contents of the Capsules to a suitable volumetric flask. Wash the blade with dehydrated alcohol, and transfer the washings to the volumetric flask. Dilute the contents of the volumetric flask with dehydrated alcohol to volume. Dilute a volume of this solution with dehydrated alcohol to obtain a solution having a nominal concentration of 1 mg/mL of cyclosporine. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L16 Column temperature: 70 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Capacity factor, k: 310 Column efficiency: NLT 700 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C62H111N11O12 in each Capsule taken: Result = (rU/rS) (CS/CU) 100
USP 32
= peak response area of cyclosporine from the Sample solution = peak response area of cyclosporine from the rS Standard solution = concentration of USP Cyclosporine RS in the CS Standard solution (mg/mL) = nominal concentration of cyclosporine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PROCEDURE FOR POWDER CAPSULES Mobile phase: Acetonitrile, methanol, water, and phosphoric acid (605:50:400:0.5) Diluent: Acetonitrile, tetrahydrofuran, and dehydrated alcohol (9:5:4) Standard solution: 25 mg of USP Cyclosporine RS in a 25mL volumetric flask. Add 2.5 mL of water, and sonicate for 10 min. Add 10 mL of Diluent, sonicate for 5 min, and dilute with Diluent to volume. Sample stock solution: Transfer the contents of 20 Capsules to a volumetric flask of such capacity, V, in mL, to make a final concentration of nominally 10 mg/mL of cyclosporine. Add 0.1V mL of water to the flask, and sonicate for 10 min. Add 0.4V mL of Diluent to the flask, and sonicate for 5 min. Dilute with Diluent to volume. Sample solution: Transfer 5.0 mL of Sample stock solution to a 50-mL volumetric flask, add 5 mL of water, and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L13 Column temperature: 70 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 700 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C62H111N11O12 in each Capsule taken: Result = (rU/rS) (CS/CU) 100 = peak response area of cyclosporine from the Sample solution = peak response area of cyclosporine from the rS Standard solution = concentration of USP Cyclosporine RS in the CS Standard solution (mg/mL) = nominal concentration of cyclosporine in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU PERFORMANCE TESTS DISSOLUTION 711 Liquid capsules Medium: Water; 500 mL Apparatus 2: 50 rpm Time: 15 min Analysis: Place 1 Capsule in each vessel, and allow the Capsule to sink to the bottom of the vessel before starting rotation of the blade. Observe the Capsules, and record the time taken for each Capsule shell to rupture. rU

Tolerances: The requirements are met if all of the Capsules tested rupture in NMT 15 min. If 1 or 2 of the Capsules rupture in NLT 15 min, but NMT 30 min, repeat the test on 12 additional Capsules. NMT 2 of the total of 18 Capsules tested rupture in NLT 15 min, but NMT 30 min. Powder capsules Medium: 0.1 N hydrochloric acid containing 0.5% of sodium lauryl sulfate; 1000 mL Apparatus 1: 150 rpm Time: 90 min Mobile phase: Acetonitrile, methanol, water, and phosphoric acid (900:50:450:0.5) Standard solution: 0.001L mg/mL of USP Cyclosporine RS in Medium. [NOTE(L stands for labeled quantity, in mg, of cyclosporine in each Capsule.] Transfer 25.0 mL of this solution to a 50-mL volumetric flask, dilute with acetonitrile to volume, and mix to obtain a concentration of 0.0005L mg/mL of USP Cyclosporine RS. Sample solution: Filter a portion of the solution under test. Transfer 5.0 mL of the filtrate to a 10-mL volumetric flask, dilute with acetonitrile to volume, and mix. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L1 Column temperature: 80 Flow rate: 2 mL/min Injection size: 10 L of the solution estimated to contain 0.1 mg/mL of cyclosporine, or 40 L of the solution estimated to contain 0.025 mg/mL of cyclosporine of the Standard solution and Sample solution System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 700 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C62H111N11O12 dissolved: Result = (rU/rS) CS V D (100/L) = peak response area of cyclosporine from the Sample solution = peak response area of cyclosporine from the rS Standard solution CS = concentration of USP Cyclosporine RS in the Standard solution (mg/mL) V = volume of Medium (mL), 1000 D = dilution factor of the Sample solution, 2 L = capsule label claim (mg) Tolerances: NLT 80% (Q) of the labeled amount of C62H111N11O12 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU SPECIFIC TESTS WATER DETERMINATION, Method I 921: NMT 3.5% for Capsules that contain powder, using finely ground Capsule contents ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, and store at controlled room temperature. USP REFERENCE STANDARDS 11 USP Cyclosporine RS
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USP 32
Sample solution 2 (where the label states the quantity of cyclosporine in a given volume): nominally 0.5 mg/mL of cyclosporine from Injection diluted with methanol [NOTE Use this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L16 Column temperature: 70 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 3NMT 10 Column efficiency: NLT 700 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of label claim of C62H111N11O12 of Injection taken: Result = (rU/rS) (CS/CU) 100 = cyclosporine peak response from the Sample solution = cyclosporine peak response from the Standard rS solution = concentration of the Standard solution (mg/mL) CS = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% rU OTHER COMPONENTS ALCOHOL CONTENT (where present) Internal standard solution: n-Propyl alcohol and butyl alcohol (3:50) Standard stock solution: 64 mg/mL of dehydrated alcohol in butyl alcohol Standard solution: Transfer 5.0 mL of Standard stock solution and 6.0 mL of Internal standard solution to a 25-mL volumetric flask, and dilute with butyl alcohol to volume. Sample solution: Transfer a portion of Injection, equivalent to 320 mg of C2H5OH, to a 25-mL volumetric flask. Add 6.0 mL of Internal standard solution, and dilute with butyl alcohol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 2-m glass column packed with support S3 Temperature: Column See the gradient table below.
Cyclosporine Injection
(Comment on this Monograph)id=m21483=Cyclosporine Injection=Chl-Cy-Monos.pdf) DEFINITION Cyclosporine Injection is a sterile solution of Cyclosporine in a suitable vehicle. It contains NLT 90.0% and NMT 110.0% of the labeled amount of cyclosporine (C62H111N11O12). IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 0.5 mg/mL of USP Cyclosporine RS in methanol Sample solution: 0.5 mg/mL of cyclosporine in methanol Solution A: 17 mg/mL of bismuth subnitrate in 20% acetic acid Solution B: 400 mg/mL of potassium iodide Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system 1: Ethyl ether Developing solvent system 2: Ethyl acetate, methyl ethyl ketone, water, and formic acid (60:40:2:1) Spray reagent 1: Solution A, Solution B, glacial acetic acid, and water (1:1:4:14) [NOTEPrepare fresh.] Spray reagent 2: Hydrogen peroxide TS Analysis Samples: Standard solution and Sample solution Proceed as directed under General chapter. Allow the spots to dry in a current of air, place the plate in a suitable chromatographic chamber, and develop the chromatogram, using Developing solvent system 1, until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow it to dry. Place the plate in a second chromatographic chamber, and develop the chromatogram in Developing solvent system 2 until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, and allow it to dry. Spray the plate with Spray reagent 1. Immediately again spray the plate with Spray reagent 2. Cyclosporine appears as a brown spot having an RF value of 0.45 on the chromatograms. [NOTEDisregard any spots at the origin.] Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, water, and phosphoric acid (550:50:400:0.5) Standard solution: 0.5 mg/mL of USP Cyclosporine RS in methanol [NOTEUse this solution promptly after preparation.] Sample solution 1 (where it is represented as being in a single-dose container): Withdraw all of the withdrawable contents from 1 container of Injection, and dilute with methanol to obtain a solution containing nominally 0.5 mg/mL of cyclosporine. [NOTEUse this solution promptly after preparation.]
USP 32
Injector: 280 Detector: 290 Carrier gas: Nitrogen Flow rate: 35 mL/min Injection size: 1 L [NOTEMake adjustments, if necessary, to obtain satisfactory chromatography.] Elution order: Alcohol, n-propyl alcohol, and butyl alcohol System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C2H5OH in the portion of Injection taken: Result = (RU/RS) (CS/CU) 100 = peak area ratio of the alcohol peak to the npropyl alcohol internal standard peak from the Sample solution = peak area ratio of the alcohol peak to the nRS propyl alcohol internal standard peak from the Standard solution = concentration of the Standard solution (mg/mL) CS = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 80.0%120.0% of labeled amount RU SPECIFIC TESTS BACTERIAL ENDOTOXINS TEST 85 Sample solution: Using USP Endotoxin RS, make a 1:10 dilution of the Injection with Water for Injection. Add 0.1 mL of the resulting suspension and 0.1 mL of appropriately constituted LAL reagent to a suitable pyrogen-free test tube. Mix on a vortex mixer for about 5 s. Acceptance criteria: The article under test contains NMT 0.84 USP Endotoxin Unit/mg of cyclosporine. STERILITY TESTS 71: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers. LABELING: Label it to indicate that it is to be diluted with a suitable parenteral vehicle prior to intravenous infusion. USP REFERENCE STANDARDS 11 USP Cyclosporine RS USP Endotoxin RS

Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system 1: Ethyl ether Developing solvent system 2: Ethyl acetate, methyl ethyl ketone, water, and formic acid (60:40:2:1) Spray reagent 1: Solution A, Solution B, glacial acetic acid, and water (1:1:4:14) [NOTEPrepare freshly.] Spray reagent 2: Hydrogen peroxide TS Analysis Samples: Standard solution and Sample solution Procedure: Proceed as directed for Chromatography 621, Thin-Layer Chromatography. Allow the spots to dry in a current of air, place the plate in a suitable chromatographic chamber, and develop the chromatogram using Developing solvent system 1 until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the chamber, mark the solvent front, and allow it to dry. Place the plate in a second chromatographic chamber, and develop the chromatogram in Developing solvent system 2 until the solvent front has moved about threefourths of the length of the plate. Remove the plate from the chamber, and allow it to dry. Spray the plate with Spray reagent 1. Immediately again spray the plate with Spray reagent 2. Cyclosporine appears as a brown spot having an RF value of 0.45 on the chromatograms. [NOTE Disregard any spots at the origin.] Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. The retention time of the major peak from the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, water, and phosphoric acid (550:50:400:0.5) Diluent: Methanol and chloroform (4:1) Standard solution: 1 mg/mL of USP Cyclosporine RS in Diluent. [NOTEUse this solution promptly after preparation.] Sample solution: nominally 1 mg/mL of cyclosporine from Oral Solution diluted with Diluent. [NOTEUse this solution promptly after preparation.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 4.6-mm 25-cm; packing L16 Column temperature: 50 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 3NMT 10 Column efficiency: NLT 700 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C62H111N11O12 in each mL of the Oral Solution taken: Result = (rU/rS) (CS/CU) 100 rU rS = cyclosporine peak response from the Sample solution = cyclosporine peak response from the Standard solution
Cyclosporine Oral Solution

(Comment on this Monograph)id=m21488=Cyclosporine Oral Solution=Chl-Cy-Monos.pdf) DEFINITION Cyclosporine Oral Solution is a solution of Cyclosporine in a suitable vehicle. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C62H111N11O12. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHY Standard solution: 1 mg/mL of USP Cyclosporine RS in a solution of methanol and chloroform (4:1) Sample solution: 1 mg/mL of cyclosporine in a solution of methanol and chloroform (4:1) Solution A: 17 mg/mL of bismuth subnitrate in 20% acetic acid Solution B: 400 mg/mL of potassium iodide Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
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CS CU
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cyclosporine RS
= concentration of the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% OTHER COMPONENTS ALCOHOL CONTENT (where present) Internal standard solution: n-propyl alcohol and butyl alcohol (3:50) Standard stock solution: 50 mg/mL of dehydrated alcohol in butyl alcohol Standard solution: Transfer 5.0 mL of Standard stock solution and 6.0 mL of Internal standard solution to a 25-mL volumetric flask, and dilute with butyl alcohol to volume. Sample solution: Transfer a portion of Oral Solution equivalent to 250 mg of C2H5OH to a 25-mL volumetric flask, add 6.0 mL of Internal standard solution, and dilute with butyl alcohol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 2-m glass column packed with support S3 Temperature Column: See the gradient table below.
Cyproheptadine Hydrochloride
(Comment on this Monograph)id=m21670=Cyproheptadine Hydrochloride=Chl-Cy-Monos.pdf)
C21H21N HCl 11/2H2O 350.88 C21H21N HCI 323.87 Piperidine, 4-(5H-dibenzo[a,d]cyclohepten-5-ylidene)-1-methyl-, hydrochloride, sesquihydrate; 4-(5H-Dibenzo[a,d]cyclohepten-5-ylidene)-1-methylpiperidine hydrochloride sesquihydrate [41354-29-4]. Anhydrous [969-33-5]. DEFINITION Cyproheptadine Hydrochloride contains NLT 98.5% and NMT 100.5% of C21H21N HCl, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 286 nm Medium: Alcohol Sample solution: 16 g/mL Acceptance criteria: Absorptivities do not differ by more than 3.0%. C. PROCEDURE Sample solution: 10 mg/mL of Cyproheptadine Hydrochloride in methanol Analysis: Place 1 drop of the solution on a filter paper, dry, and view under short-wavelength UV light. Acceptance criteria: A bright blue fluorescence is observed. ASSAY PROCEDURE Sample solution: 650 mg of Cyproheptadine Hydrochloride in 50 mL of glacial acetic acid, heating to dissolve. Cool, and add 10 mL of mercuric acetate TS, 0.5 mL of acetic anhydride, and 1 drop of crystal violet TS. Analysis: Titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination, and make any necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 32.39 mg of C21H21N HCl. Acceptance criteria: 98.5%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: 30 ppm SPECIFIC TESTS ACIDITY: 1.0 g of Cyproheptadine Hydrochloride in 25 mL of methanol. Add methyl red TS, and titrate with 0.10 N sodium hydroxide: NMT 0.15 mL is required (0.05% as HCl).
Injector: 280 Detector: 290 Carrier gas: Nitrogen Flow rate: 35 mL/min Injection size: 1 L [NOTEMake adjustments if necessary to obtain satisfactory chromatography.] Elution order: Alcohol, n-propyl alcohol, and butyl alcohol System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C2H5OH in the portion of Oral Solution taken: Result = (RU/RS) (CS/CU) 100 = peak area response ratio of the alcohol peak to the n-propyl alcohol internal standard peak from the Sample solution = peak area response ratio of the alcohol peak to RS the n-propyl alcohol internal standard peak from the Standard solution CS = concentration of the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) Acceptance criteria: 80.0%120.0% of labeled amount RU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For oral solution packaged in single-unit containers: meets the requirements. DELIVERABLE VOLUME 698: Meets the requirements for oral solution packaged in multiple-unit containers
USP 32
WATER DETERMINATION, Method I 921: 7.0%9.0% rS CS
Official Monographs / Cyproheptadine 221

= peak response from the Standard solution = concentration of USP Cyproheptadine Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of cyproheptadine hydrochloride (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 3.54.5 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Cyproheptadine Hydrochloride RS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cyproheptadine Hydrochloride RS
Cyproheptadine Hydrochloride Oral Solution

(Comment on this Monograph)id=m21700=Cyproheptadine Hydrochloride Oral Solution=Chl-Cy-Monos.pdf) DEFINITION Cyproheptadine Hydrochloride Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of C21H21N HCl IDENTIFICATION PROCEDURE Solution A: Sodium bicarbonate (2 in 100) Sample solution: Place 50 mL of Oral Solution in a separator, add 25 mL of Solution A, and extract with three 15-mL portions of isooctane. Wash the combined isooctane extracts with 15 mL of Solution A, and discard the washings. Evaporate the isooctane solution on a steam bath to dryness, and dissolve the residue in 1 mL of carbon disulfide, filtering if necessary. Analysis: Determine the IR absorption spectrum as directed under IdentificationOrganic Nitrogenous Bases 181, obtaining the spectrum of USP Cyproheptadine Hydrochloride RS as directed. Acceptance criteria: The Oral Solution meets the requirements of the test. ASSAY PROCEDURE Solution A: Methanesulfonic acid in water (3:1000) Mobile phase: Acetonitrile, isopropyl alcohol, and Solution A (4:3:13); while mixing, adjust with triethylamine to a pH of 4.0 0.05. Standard solution: 0.02 mg/mL of USP Cyproheptadine Hydrochloride RS in Mobile phase Sample solution: Transfer a volume of Oral Solution, nominally equivalent to 2 mg of cyproheptadine hydrochloride to a 100-mL volumetric flask. Dilute with Mobile phase to volume, and mix. Pass the solution through a filter having a 0.45-m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 285 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 L/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H21N HCl in the portion of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 rU = peak response from the Sample solution
Cyproheptadine Hydrochloride Tablets

(Comment on this Monograph)id=m21710=Cyproheptadine Hydrochloride Tablets=Chl-Cy-Monos.pdf) DEFINITION Cyproheptadine Hydrochloride Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C21H21N HCl. IDENTIFICATION IDENTIFICATIONORGANIC NITROGENOUS BASES 181: the requirements Meet
ASSAY PROCEDURE Solution A: Methanesulfonic acid in water (3:1000) Mobile phase: Acetonitrile, isopropyl alcohol, and Solution A (4:3:13). While mixing, adjust with triethylamine to a pH of 4.0 0.05. Filter, and degas. Standard solution: 0.08 mg/mL of USP Cyproheptadine Hydrochloride RS in Mobile phase Sample solution: Transfer a number of Tablets, nominally equivalent to 80 mg of cyproheptadine hydrochloride to a 1-L volumetric flask, dissolve by sonication in 500 mL of Mobile phase for 15 min, and agitate for 30 min. Dilute with Mobile phase to volume. Pass through a filter having a 0.45m or finer porosity. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 285 nm Column: 3.9-mm 15-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H21N HCl in each of the Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cyproheptadine Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of cyproheptadine hydrochloride in the Sample solution (mg/mL)
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Cyproheptadine / Official Monographs

90.0%110.0%
USP 32
Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 214 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 3000 theoretical plates Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H10N6 in the portion of Cyromazine taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cyromazine RS in the Standard solution (g/mL) CU = concentration of Cyromazine in the Sample solution (g/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281:
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Dilute with Medium as needed. Standard solution: USP Cyproheptadine Hydrochloride RS at a known concentration in Medium Spectrometric conditions: Mode: UV Analytical wavelength: 285 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 80% (Q) of the labeled amount of C21H21N HCl is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Cyproheptadine Hydrochloride RS
Cyromazine
(Comment on this Monograph)id=m2041=Cyromazine=Chl-CyMonos.pdf)
NMT 0.1%
SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 219226 LOSS ON DRYING 731: Dry it at 105 to a constant weight: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Cyromazine RS
166.18 C6H10N6 N-Cyclopropyl-1,3,5-triazine-2,4,6-triamine; 2-Cyclopropylamino-4,6-diamino-s-triazine [66215-27-8]. DEFINITION Cyromazine contains NLT 98.0% and NMT 102.0% of C6H10N6, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Mix 930 mL of water, 3.72 g of dibasic potassium phosphate, and 6.48 g of monobasic potassium phosphate. Add 50 mL of methanol and 20 mL of acetonitrile. Standard stock solution: 0.50 mg/mL of USP Cyromazine RS in methanol Standard solution: 10 g/mL of USP Cyromazine RS from Standard stock solution diluted with Mobile phase Sample stock solution: 0.50 mg/mL of cyromazine in methanol Sample solution: 10 g/mL of Cyromazine from Sample stock solution diluted with Mobile phase.
Cysteine Hydrochloride
(Comment on this Monograph)id=m21760=Cysteine Hydrochloride=Chl-Cy-Monos.pdf)
C3H7NO2S HCl H2O L-Cysteine hydrochloride monohydrate [7048-04-6]. Anhydrous [52-89-1].
175.64 157.62
DEFINITION Cysteine Hydrochloride contains NLT 98.5% and NMT 101.5% of C3H7NO2S HCl, as L-cysteine hydrochloride, calculated on the dried basis.
USP 32
IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample solution: 250 mg of Cysteine Hydrochloride in an iodine flask. Add 20 mL of water and 4 g of potassium iodide, and mix. Cool the solution in an ice bath, and add 5 mL of 3 N hydrochloric acid and 25.0 mL of 0.1 N iodine VS. Insert the stopper, and allow to stand in the dark for 20 min. Analysis: Titrate the excess iodine with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as the endpoint is approached. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N sodium thiosulfate is equivalent to 15.76 mg of C3H7NO2S HCl. Acceptance criteria: 98.5%101.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.4% CHLORIDE AND SULFATE, Sulfate 221: A solution containing 0.33 g shows no more sulfate than corresponds to 0.10 mL of 0.020 N sulfuric acid (0.03%). IRON 241: NMT 30 ppm HEAVY METALS, Method I 231: NMT 15 ppm Organic Impurities PROCEDURE Solution A: 4% N-ethylmaleimide in alcohol (w/v) Standard stock solution: 20 mg of USP L-Cysteine Hydrochloride RS in 10.0 mL of water. Add 10.0 mL of Solution A. Allow the solution to stand for 5 min before using. Standard solution: 0.05 mg/mL from Standard stock solution in water [NOTEThis solution has a concentration equivalent to 0.5% of that of the Sample solution.] System suitability solution: 10 mg of USP L-Tyrosine RS and 10 mL of the Standard stock solution in a 25-mL volumetric flask. Dilute with water to volume. Sample solution: Transfer 0.2 g of Cysteine Hydrochloride to a 10-mL volumetric flask, dissolve, and dilute with water to volume. To 5.0 mL of this solution, add 5.0 mL of Solution A. [NOTEAllow the solution to stand for 5 min before use.] Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Butyl alcohol, glacial acetic acid, and water (3:1:1) Spray reagent: 2 mg/mL of ninhydrin in a mixture of butyl alcohol and 2 N acetic acid (95:5) Analysis Samples: Standard solution and Sample solution Proceed as directed for Chromatography 621, Thin-Layer Chromatography. After air-drying the plate, spray with Spray reagent, and heat between 100 and 105 for about 15 min. Examine the plate under white light. The chromatogram of the System suitability solution exhibits two clearly separated spots. Acceptance criteria Any individual impurity: Any secondary spot in the chromatogram of the Sample solution is not larger or more intense than the principal spot in the chromatogram of the Standard solution. NMT 0.5%
Official Monographs / Cysteine 223

Total impurities: NMT 2.0%
SPECIFIC TESTS SPECIFIC ROTATION 781S: +5.7 to +6.8 Sample solution: 80 mg/mL, in 6 N hydrochloric acid LOSS ON DRYING 731: Dry it at a pressure not exceeding 5 mm of mercury for 24 h: it loses between 8.0% and 12.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP L-Cysteine Hydrochloride RS USP L-Tyrosine RS
Cysteine Hydrochloride Injection

(Comment on this Monograph)id=m21770=Cysteine Hydrochloride Injection=Chl-Cy-Monos.pdf) DEFINITION Cysteine Hydrochloride Injection is a sterile solution of Cysteine Hydrochloride in Water for Injection. It contains NLT 85.0% and NMT 115.0% of the labeled amount of C3H7NO2S HCl H2O. IDENTIFICATION A. PROCEDURE Sample solution: To 2 mL of Injection add 3 mL of water, and mix. Analysis: Add 10 mL of cupric sulfate TS Acceptance criteria: A bluish-gray precipitate is formed. B. PROCEDURE Sample solution: To 2 mL of Injection add 3 mL of water, and mix. Analysis: Add 2 mL of 3 N sodium hydroxide and 2 drops of sodium nitroferricyanide solution (1 in 20) Acceptance criteria: A red-purple color is produced, and it rapidly changes to yellow. ASSAY PROCEDURE Standard stock solution: 1 mg/mL of USP L-Cysteine Hydrochloride RS in nitrogen-saturated water Standard solution: Transfer 20.0 mL of Standard stock solution to a 200-mL volumetric flask, dilute with nitrogensaturated 1.0 N sodium hydroxide to volume, and mix. Sample solution: Nominally equivalent to 250 mg of cysteine hydrochloride from Injection in nitrogen-saturated 1.0 N sodium hydroxide, to obtain a solution having a concentration of nominally 0.1 mg/mL Analysis Samples: Standard solution and Sample solution Transfer a suitable amount of Standard solution to a polarographic cell. With mercury dropping from the electrode, lower the dropping mercury electrode of a polarograph so that the end is submerged in the liquid. Bubble oxygen-free, water-saturated nitrogen through the liquid for 15 min. Record the polarogram from 0.2 volts to 1.10 volts, using a saturated calomel electrode as the reference electrode. In a similar manner, record the polarograms obtained using portions of the Sample solution and of the nitrogen-saturated 1.0 N sodium hydroxide. Determine the height of the diffusion current wave at 0.4 volts.
224
Cysteine / Official Monographs
USP 32
Sample solution: 0.1 mg/mL of Cytarabine in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for cytarabine and uracil arabinoside are about 1.0 and 1.3, respectively.] Suitability requirements Resolution: NLT 2.5 between cytarabine and uracil arabinoside, System suitability solution Relative standard deviation: NMT 2.0% for replicate injections, Standard solution [NOTEAfter chromatography has been completed, flush the column with a mixture of water and methanol (7:3).] Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H13N3O5 in the portion of Cytarabine taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Cytarabine RS in the Standard solution (mg/mL) = concentration of Cytarabine in the Sample CU solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.5% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE Phosphate buffer: Prepare a solution containing 0.01 M monobasic sodium phosphate and 0.01 M dibasic sodium phosphate in a suitable container. Adjust with 0.1 M sodium hydroxide or 0.1 M phosphoric acid to a pH of 7.0. Solution A: Methanol and Phosphate buffer (1:49) [NOTE Prepare this solution fresh daily.] Solution B: Methanol and Phosphate buffer (3:7) [NOTE Prepare this solution fresh daily.] Mobile phase: Use variable mixtures of Solution A and Solution B as directed under Chromatographic system.
Calculate the percentage of C3H7NO2S HCl H2O in each mL of Injection taken: Result = (CS/CU)[(id)U/(id)S] 100 = concentration of USP L-Cysteine Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of L-cysteine CU hydrochloride in the Sample solution (mg/mL) = observed diffusion current, corrected for the (id)U diffusion current of the 0.1 N sodium hydroxide, of the Sample solution = observed diffusion current, corrected for the (id)S diffusion current of the 0.1 N sodium hydroxide, of the Standard solution Acceptance criteria: 85.0%115.0% CS IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231:
NMT 2 ppm
SPECIFIC TESTS PH 791: 1.02.5 BACTERIAL ENDOTOXINS TEST 85: NMT 0.7 USP Endotoxin Unit/mg of cysteine hydrochloride OTHER REQUIREMENTS: Meets the requirements under Injections 1 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP L-Cysteine Hydrochloride RS USP Endotoxin RS
Cytarabine
(Comment on this Monograph)id=m21810=Cytarabine=Chl-CyMonos.pdf)
C9H13N3O5 2(1H)-Pyrimidinone, 4-amino-1--D-arabinofuranosyl-; 1--D-Arabinofuranosylcytosine [147-94-4]. DEFINITION Cytarabine contains NLT 98.0% and NMT 102.0% of C9H13N3O5, calculated on the dried basis.
243.22
IDENTIFICATION A. INFRARED ABSORPTION 197M: Previously dried at a pressure of NMT 5 mm of mercury at 60 for 3 h B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.73 mg/mL of monobasic sodium phosphate and 1.4 mg/mL of dibasic sodium phosphate Mobile phase: Methanol and Solution A (5:95) Standard solution: 0.1 mg/mL of USP Cytarabine RS in water System suitability solution: 0.1 mg/mL of USP Uracil Arabinoside RS in Standard solution
System suitability solution: 0.02 mg/mL of uridine, 0.02 mg/mL of USP Uracil Arabinoside RS, and 5.0 mg/mL of USP Cytarabine RS Standard solution: 4 g/mL of USP Cytarabine RS Sample solution: 5 mg/mL of Cytarabine [NOTEPrepare this solution fresh daily.] Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for uracil, uridine, uracil arabinoside, and cytarabine are about 0.55, 1.14, 1.62, and 1.0, respectively.] Suitability requirements Resolution: NLT 1.25 between cytarabine and uridine, System suitability solution Relative standard deviation: NMT 3.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of uracil arabinoside in the portion of Cytarabine taken: Result = (rU/rS) (CS/CU) (1/F) 100 = peak response of uracil arabinoside from the Sample solution = peak response of USP Cytarabine RS from the rS Standard solution = concentration of USP Cytarabine RS in the CS Standard solution (mg/mL) CU = concentration of Cytarabine in the Sample solution (mg/mL) F = relative response factor for uracil arabinoside, 1.34 Acceptance criteria: NMT 0.30% Calculate the percentage of all other impurities in the portion of Cytarabine taken: rU Result = (rU/rS) (CS/CU) (1/F) 100 = peak response of each impurity from the Sample solution = peak response of USP Cytarabine RS from the rS Standard solution CS = concentration of the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU F = relative response factor (see Impurity Table) Acceptance criteria Individual impurities: See Impurity Table 1. Total impurities (including uracil arabinoside): NMT 0.30% rU
Impurity Table 1 Relative Retention Time 1.62 0.55 0.38 0.43 1.14 Relative Response Factor 1.34 2.5 1.5 1.5 1.5 1.0 Acceptance Criteria, NMT (%) 0.30 0.10 0.10 0.10 0.10 0.10
Official Monographs / Cytarabine 225

SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +154 to +160 Sample solution: 10 mg/mL LOSS ON DRYING 731: Dry it in a vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 1.0% of its weight. STERILITY TESTS 71: Where the label states that Cytarabine is sterile, it meets the requirements. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Cytarabine is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.07 USP Endotoxin Unit/mg of cytarabine. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Cytarabine RS USP Endotoxin RS USP Uracil Arabinoside RS
Cytarabine for Injection

(Comment on this Monograph)id=m21815=Cytarabine for Injection=Chl-Cy-Monos.pdf) DEFINITION Cytarabine for Injection contains NLT 90.0% and NMT 110.0% of the labeled amount of cytarabine (C9H13N3O5). IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: 0.73 mg/mL of monobasic sodium phosphate and 1.4 mg/mL of dibasic sodium phosphate in water Mobile phase: Filtered and degassed mixture of methanol and Solution A (1:19) Standard solution: 0.1 mg/mL of USP Cytarabine RS in water System suitability solution: 0.1 mg/mL of USP Uracil Arabinoside RS in Standard solution Sample solution: Separately constitute 5 vials of Injection in a volume of water corresponding to the volume specified in the labeling. Pool and mix the constituted solutions in a suitable container. Transfer a volume of the constituted solution, nominally equivalent to about 100 mg of cytarabine, to a 100-mL volumetric flask, and dilute with water to volume. Transfer 5.0 mL of this solution to a 50-mL volumetric flask, and dilute with water to volume. Chromatographic system (See Chromatography 621, System Suitability.)
Name Uracil arabinoside Uracil Any other individual unidentified impurity Any other individual unidentified impurity Uridine Any individual impurity
226
Cytarabine / Official Monographs

CS CU
USP 32
= concentration of the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 4.06.0, in a solution containing the equivalent of 10 mg/mL of cytarabine WATER DETERMINATION, Method I 921: NMT 3.0% BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.07 USP Endotoxin Unit/mg of cytarabine. CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Constituted Solutions under Injections 1. OTHER REQUIREMENTS: It meets the requirements for Sterility Tests 71, Uniformity of Dosage Units 905, and Labeling under Injections 1. The drug substance in the vial meets the requirements for Cytarabine. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Cytarabine RS USP Endotoxin RS USP Uracil Arabinoside RS
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution and System suitability solution Suitability requirements [NOTEThe relative retention times for cytarabine and uracil arabinoside are about 1.0 and 1.3, respectively.] Resolution: NLT 2.5 between cytarabine and uracil arabinoside Relative standard deviation: NMT 2.0% determined from replicate injections of the Standard solution [NOTEAfter chromatography has been completed, flush the column with a mixture of water and methanol (7:3).] Analysis Samples: Standard solution and Sample solution Calculate the percentage of C9H13N3O5 in the portion of constituted solution taken: Result = (rU/rS) (CS/CU) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
USP 32
Dacarbazine Dacarbazine for Injection Dactinomycin Dactinomycin for Injection Danazol Danazol Capsules Dantrolene Sodium Dantrolene Sodium Capsules Dantrolene Sodium for Injection Dapsone Dapsone Tablets Daunorubicin Hydrochloride Daunorubicin Hydrochloride for Injection Decoquinate Decoquinate Premix Deferoxamine Mesylate Deferoxamine Mesylate for Injection Dehydrocholic Acid Dehydrocholic Acid Tablets Demecarium Bromide Demecarium Bromide Ophthalmic Solution Demeclocycline Demeclocycline Oral Suspension Demeclocycline Hydrochloride Demeclocycline Hydrochloride Capsules Demeclocycline Hydrochloride Tablets Cryopreserved Human Fibroblast-Derived Dermal Substitute Desflurane Desipramine Hydrochloride Desipramine Hydrochloride Tablets Deslanoside Deslanoside Injection Desmopressin Acetate Desmopressin Acetate Injection Desogestrel and Ethinyl Estradiol Tablets Desoximetasone Desoximetasone Cream Desoximetasone Gel Desoximetasone Ointment Desoxycorticosterone Acetate Desoxycorticosterone Acetate Injection Desoxycorticosterone Acetate Pellets Desoxycorticosterone Pivalate
MONOGRAPH LIST /
Desoxycorticosterone Pivalate Injectable Suspension Dexamethasone Dexamethasone Topical Aerosol Dexamethasone Elixir Dexamethasone Gel Dexamethasone Injection Dexamethasone Ophthalmic Suspension Dexamethasone Oral Solution Dexamethasone Tablets Dexamethasone Acetate Dexamethasone Acetate Injectable Suspension Dexamethasone Sodium Phosphate Dexamethasone Sodium Phosphate Inhalation Aerosol Dexamethasone Sodium Phosphate Cream Dexamethasone Sodium Phosphate Injection Dexamethasone Sodium Phosphate Ophthalmic Ointment Dexamethasone Sodium Phosphate Ophthalmic Solution Dexbrompheniramine Maleate
Dexbrompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution Dexchlorpheniramine Maleate Dexchlorpheniramine Maleate Oral Solution Dexchlorpheniramine Maleate Tablets Dexpanthenol Dexpanthenol Preparation Dextran 1 Dextran 40 Dextran 40 in Dextrose Injection Dextran 40 in Sodium Chloride Injection Dextran 70 Dextran 70 in Dextrose Injection Dextran 70 in Sodium Chloride Injection Dextroamphetamine Sulfate Dextroamphetamine Sulfate Capsules Dextroamphetamine Sulfate Tablets Dextromethorphan Dextromethorphan Hydrobromide Dextromethorphan Hydrobromide Oral Solution Dextrose Dextrose Injection Dextrose and Sodium Chloride Injection Diatrizoate Meglumine Diatrizoate Meglumine Injection
/ MONOGRAPH LIST
Diethylpropion Hydrochloride Diethylpropion Hydrochloride Tablets Diethylstilbestrol Diethylstilbestrol Injection Diethylstilbestrol Tablets Diethylstilbestrol Diphosphate Diethyltoluamide Diethyltoluamide Topical Solution Diflorasone Diacetate Diflorasone Diacetate Cream Diflorasone Diacetate Ointment Diflunisal Diflunisal Tablets Digitalis Powdered Digitalis Digitalis Capsules Digitalis Tablets Digitoxin Digitoxin Injection Digitoxin Tablets Digoxin Digoxin Injection Digoxin Oral Solution Digoxin Tablets Dihydrocodeine Bitartrate Dihydroergotamine Mesylate Dihydroergotamine Mesylate Injection Dihydrostreptomycin Sulfate Dihydrostreptomycin Sulfate Boluses Dihydrostreptomycin Injection Dihydrotachysterol Dihydrotachysterol Capsules Dihydrotachysterol Oral Solution Dihydrotachysterol Tablets Dihydroxyacetone Dihydroxyaluminum Aminoacetate Dihydroxyaluminum Aminoacetate Magma Dihydroxyaluminum Sodium Carbonate Dihydroxyaluminum Sodium Carbonate Tablets
USP 32
Diatrizoate Meglumine and Diatrizoate Sodium Injection Diatrizoate Meglumine and Diatrizoate Sodium Solution Diatrizoate Sodium Diatrizoate Sodium Injection Diatrizoate Sodium Solution Diatrizoic Acid Diazepam Diazepam Capsules Diazepam Extended-Release Capsules Diazepam Injection Diazepam Tablets Diazoxide Diazoxide Capsules Diazoxide Injection Diazoxide Oral Suspension Dibucaine Dibucaine Cream Dibucaine Ointment Dibucaine Hydrochloride Dibucaine Hydrochloride Injection Dichloralphenazone Dichlorphenamide Dichlorphenamide Tablets Diclofenac Potassium Diclofenac Potassium Tablets Diclofenac Sodium Diclofenac Sodium Delayed-Release Tablets Diclofenac Sodium Extended-Release Tablets Dicloxacillin Sodium Dicloxacillin Sodium Capsules Dicloxacillin Sodium for Oral Suspension Dicyclomine Hydrochloride Dicyclomine Hydrochloride Capsules Dicyclomine Hydrochloride Injection Dicyclomine Hydrochloride Oral Solution Dicyclomine Hydrochloride Tablets Didanosine Didanosine for Oral Solution Didanosine Tablets for Oral Suspension Dienestrol Dienestrol Cream Diethylcarbamazine Citrate Diethylcarbamazine Citrate Tablets
Dihydroxyaluminum Sodium Carbonate Chewable Tablets Diloxanide Furoate Diltiazem Hydrochloride Diltiazem Hydrochloride Extended-Release Capsules
USP 32
Diltiazem Hydrochloride Oral Solution Diltiazem Hydrochloride Oral Suspension Diltiazem Hydrochloride Tablets Dimenhydrinate Dimenhydrinate Injection Dimenhydrinate Oral Solution Dimenhydrinate Tablets Dimercaprol Dimercaprol Injection Dimethyl Sulfoxide Dimethyl Sulfoxide Gel Dimethyl Sulfoxide Irrigation Dimethyl Sulfoxide Topical Solution Dinoprost Tromethamine Dinoprost Tromethamine Injection Dinoprostone Dioxybenzone Dioxybenzone and Oxybenzone Cream Diphenhydramine Citrate Diphenhydramine Hydrochloride Diphenhydramine Hydrochloride Capsules Diphenhydramine Hydrochloride Injection Diphenhydramine Hydrochloride Oral Solution Diphenhydramine and Pseudoephedrine Capsules Diphenoxylate Hydrochloride Diphenoxylate Hydrochloride and Atropine Sulfate Oral Solution Diphenoxylate Hydrochloride and Atropine Sulfate Tablets Diphtheria and Tetanus Toxoids Adsorbed Dipivefrin Hydrochloride Dipivefrin Hydrochloride Ophthalmic Solution Dipyridamole Dipyridamole Injection Dipyridamole Oral Suspension Dipyridamole Tablets Dirithromycin Dirithromycin Delayed-Release Tablets Disopyramide Phosphate Disopyramide Phosphate Capsules Disopyramide Phosphate Extended-Release Capsules Disulfiram Disulfiram Tablets Divalproex Sodium
MONOGRAPH LIST /
Divalproex Sodium Delayed-Release Tablets Dobutamine Hydrochloride Dobutamine Injection Dobutamine for Injection Dobutamine in Dextrose Injection Docusate Calcium Docusate Calcium Capsules Docusate Potassium Docusate Potassium Capsules Docusate Sodium Docusate Sodium Capsules Docusate Sodium Solution Docusate Sodium Syrup Docusate Sodium Tablets Dolasetron Mesylate Dolasetron Mesylate Injection Dolasetron Mesylate Oral Solution Dolasetron Mesylate Oral Suspension Dolasetron Mesylate Tablets Dopamine Hydrochloride Dopamine Hydrochloride Injection Dopamine Hydrochloride and Dextrose Injection Dorzolamide Hydrochloride Doxapram Hydrochloride Doxapram Hydrochloride Injection Doxazosin Mesylate Doxazosin Tablets Doxepin Hydrochloride Doxepin Hydrochloride Capsules Doxepin Hydrochloride Oral Solution Doxorubicin Hydrochloride Doxorubicin Hydrochloride Injection Doxorubicin Hydrochloride for Injection Doxycycline Doxycycline Capsules Doxycycline for Oral Suspension Doxycycline Calcium Oral Suspension Doxycycline Hyclate Doxycycline Hyclate Capsules Doxycycline Hyclate Delayed-Release Capsules Doxycycline for Injection Doxycycline Hyclate Tablets Doxylamine Succinate
/ MONOGRAPH LIST
Dyclonine Hydrochloride Topical Solution Dydrogesterone Dydrogesterone Tablets Dyphylline Dyphylline Injection Dyphylline Oral Solution Dyphylline Tablets Dyphylline and Guaifenesin Oral Solution Dyphylline and Guaifenesin Tablets
USP 32
Doxylamine Succinate Oral Solution Doxylamine Succinate Tablets Dronabinol Dronabinol Capsules Droperidol Droperidol Injection Drospirenone Absorbable Dusting Powder Dyclonine Hydrochloride Dyclonine Hydrochloride Gel
USP 32
Official Monographs / Dacarbazine 1

= absorbance of Acidic standard solution at 323 nm AS2 = absorbance of Neutral standard solution at 329 nm CS = concentration, of USP Dacarbazine RS in the Standard solution (g/mL) = concentration, of Dacarbazine in the Sample CU solution (g/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Standard solutions: 0.40 mg/mL of USP Dacarbazine Related Compound A RS in methanol and 0.40 mg/mL of USP Dacarbazine Related Compound B RS in water. Sample solution: 40 mg/mL of Dacarbazine in 0.1 N hydrochloric acid Chromatographic system (See Chromatography 621, Thin-Layer Chromatograhy.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Butanol, acetic acid, and water (5:1:2) Analysis Samples: Standard solutions and Sample solution Locate the spots on the plate by viewing under shortwavelength UV light. Acceptance criteria: Any spots from the Sample solution are not greater in size or intensity than the spots, occurring at the respective RF values, produced by the Standard solutions, corresponding to NMT 1.0% of dacarbazine related compound A and NMT 1.0% of dacarbazine related compound B. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, in a refrigerator. USP REFERENCE STANDARDS 11 USP Dacarbazine RS USP Dacarbazine Related Compound A RS USP Dacarbazine Related Compound B RS AS1
Dacarbazine
132270
(Comment on this Monograph)id=m21850=Dacarbazine=DMonos.pdf)
C6H10N6O 182.18 1H-Imidazole-4-carboxamide, 5-(3,3-dimethyl-1-triazenyl)-; 5-(3,3-Dimethyl-1-triazeno)imidazole-4-carboxamide [4342-03-04]. DEFINITION Dacarbazine contains NLT 97.0% and NMT 102.0% of C6H10N6O. [CAUTIONGreat care should be taken in handling Dacarbazine, as it is a potent cytotoxic agent.] IDENTIFICATION The IR absorption spectrum of a potassium bromide dispersion of it exhibits maxima only at the same wavelengths as that of a similar preparation of USP Dacarbazine RS. ASSAY PROCEDURE [NOTEThroughout this procedure, avoid exposing Dacarbazine and its solutions to light.] Standard stock solution: 0.6 mg/mL of USP Dacarbazine RS in 0.1 N hydrochloric acid Acidic standard solution: 6 g/mL of Standard stock solution in 0.1 N hydrochloric acid Neutral standard solution: 6 g/mL of Standard stock solution in pH 7.0 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions) Sample stock solution: 0.6 mg/mL of Dacarbazine in 0.1 N hydrochloric acid Acidic sample solution: 6 g/mL of Sample stock solution in 0.1 N hydrochloric acid Neutral sample solution: 6 g/mL of Sample stock solution in pH 7.0 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions) Analysis 1 Samples: Blank, Acidic standard solution, and Acidic sample solution Spectrometric conditions 1 Mode: UV Analytical wavelength: 323 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis 2 Samples: Blank, Neutral standard solution, and Neutral sample solution Spectrometric conditions 2 Mode: UV Analytical wavelength: 329 nm Cell: 1 cm Blank: pH 7.0 phosphate buffer (see Reagents, Indicators, and SolutionsBuffer Solutions) Calculate the percentage of C6H10N6O in the portion of Dacarbazine taken: Result = [(AU1 + AU2) / (AS1 + AS2)](CS/CU) 100 AU1 AU2 = absorbance of Acidic sample solution at 323 nm = absorbance of Neutral sample solution at 329 nm
Dacarbazine for Injection

(Comment on this Monograph)id=m21860=Dacarbazine for Injection=D-Monos.pdf) DEFINITION Dacarbazine for Injection is a sterile, freeze-dried mixture of Dacarbazine and suitable buffers or diluents. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C6H10N6O. [CAUTIONGreat care should be taken to prevent inhaling particles of Dacarbazine for Injection and exposing the skin to it.] IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 10 mg/mL each of USP Dacarbazine RS and citric acid in water Sample solution: 10 mg/mL of Dacarbazine for Injection in water Chromatographic system (See Chromatography 621,Thin-layer Chromatography.)
Dacarbazine / Official Monographs

CU
USP 32
= nominal concentration of dacarbazine in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements IMPURITIES Organic Impurities PROCEDURE: LIMIT OF 2-AZAHYPOXANTHINE [NOTEThe Mobile phase employed in this procedure is corrosive. The system should be rinsed well with methanol following completion of analysis.] Mobile phase: 2.2 g of docusate sodium to a 1000-mL volumetric flask, dissolve in a mixture of 100 mL of water and 15 mL of glacial acetic acid, and dilute with water to volume. Pass the solution through a 0.5-m porosity filter. Prepare this solution fresh daily. Standard solution: 0.04 mg/mL of USP Dacarbazine Related Compound B RS Sample solution: 4 mg/mL of Dacarbazine for Injection in water [NOTEConstitute the contents of 1 vial of Dacarbazine for Injection. Using the contents of the constituted vial, dilute quantitatively with water.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% (5 replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percentage of 2-azahypoxanthine monohydrate in the dacarbazine taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dacarbazine Related Compound B RS in the Standard solution (mg/mL) = nominal concentration for the Sample solution CU Acceptance criteria: NMT 1.0% rU rS CS SPECIFIC TESTS PH 791: 3.04.0, in a solution containing an amount of Dacarbazine for Injection equivalent to 1 g of dacarbazine in 100 mL of water WATER, Method I 921: NMT 1.5% BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.52 USP Endotoxin Unit/mg of dacarbazine. STERILITY TESTS 71: Meets the requirements COMPLETENESS OF SOLUTION: When dissolved as directed in the labeling, it yields a clear, pale yellow to yellow solution. CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions.
Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel. Application volume: 1 L Developing solvent system: Isopropyl alcohol and 1 N ammonium hydroxide (3:1) Spray reagent: Freshly prepared solution containing 1% of ferric chloride and 1% of potassium ferricyanide (prepared by mixing 5 mL of a 10% aqueous solution of ferric chloride with 5 mL of a 10% aqueous solution of potassium ferricyanide and diluting with water to 50 mL) Analysis Samples: Standard solution and Sample solution Develop the chromatogram until the solvent front has moved three-quarters of the length of the plate. Spray the plate evenly with Spray reagent. Dacarbazine appears as an intense blue spot on a light yellow background. Acceptance criteria: The RF value of the spot from the Sample solution corresponds to that from the Standard solution. B. To 1 mL of a solution (1 in 100) in a test tube add a few crystals of periodic acid and 4 drops of methanol. Shake, and after 1 min add 5 mL of a 0.2% acetylacetone reagent solution (prepared by mixing 15.0 g of ammonium acetate, 0.30 mL of glacial acetic acid, and 0.20 mL of acetylacetone in a 100-mL volumetric flask, adding water to volume, and mixing). Shake, and place in a water bath maintained at a temperature of 60. Acceptance criteria: An intense yellow color develops in a few min (presence of mannitol). C. To 2 drops of an aqueous solution (1 in 100) in a 15-mL test tube add 10 mL of a solution prepared by mixing 10 mL of acetic anhydride with 30 mL of pyridine. Acceptance criteria: An intense yellow color is produced immediately and after a few min becomes red-violet (presence of citric acid). ASSAY PROCEDURE Sample solution: Dissolve the contents of NLT 10 containers of Dacarbazine for Injection in 0.1 N hydrochloric acid. Transfer and combine the solutions, rinsing as necessary with 0.1 N hydrochloric acid. Dilute with 0.1 N hydrochloric acid to obtain a solution containing 3.2 g/mL of dacarbazine. Standard solution: 3.2 g/mL of USP Dacarbazine RS in 0.1 N hydrochloric acid Spectrometric conditions Mode: UV Analytical wavelength: 323 nm Cell: 1 cm Blank: 0.1 N hydrochloric acid Analysis Samples: Blank, Standard solution, and Sample solution Calculate the percentage of C6H10N6O in each container of Dacarbazine for Injection taken: Result = (AU/AS) (CS/CU) 100 AU AS CS = absorbances of the Sample solution = absorbances of the Standard solution = concentration of USP Dacarbazine RS in the Standard solution (g/mL)
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose Containers for Sterile Solids as described under Injections 1, preferably of Type I glass, protected from light. LABELING: Meets requirements for Injections 1 USP REFERENCE STANDARDS 11 USP Dacarbazine RS USP Dacarbazine Related Compound B RS USP Endotoxin RS
Official Monographs / Dactinomycin 3

Analysis Samples: Standard solution and Sample solution Calculate the potency, in g, of C62H86N12O16 per mg taken: Result = (rU/rS) (CS/CU) F = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dactinomycin RS in each mL of the Standard solution (mg/mL) CU = concentration for the Sample solution (mg/mL) P = potency of USP Dactinomycin RS (g/mg) Acceptance criteria: 950 g/mg1030 g/mg rU rS CS SPECIFIC TESTS SPECIFIC ROTATION 781S: 292 to 317 (t = 20) Sample solution: 1 mg/mL, in methanol CRYSTALLINITY 695: Meets the requirements LOSS ON DRYING 731: Dry it in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 5.0% of its weight. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 100 USP Endotoxin Unit/mg ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light and excessive heat. USP REFERENCE STANDARDS 11 USP Dactinomycin RS USP Endotoxin RS
Dactinomycin
(Comment on this Monograph)id=m21890=Dactinomycin=DMonos.pdf)
C62H86N12O16 Actinomycin D [50-76-0].
1255.42
DEFINITION Dactinomycin contains NLT 950 g and NMT 1030 g of C62H86N12O16 per mg, calculated on the dried basis. [CAUTIONGreat care should be taken to prevent inhaling particles of Dactinomycin and exposing the skin to it.] IDENTIFICATION A. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 240 nm and 445 nm Sample solution: 25 g/mL in methanol Acceptance criteria: The absorptivity, calculated on the dried basis, is NLT 95.0% and NMT 103.0% of that of USP Dactinomycin RS, the potency of the Reference Standard being taken into account. Ratio: A240/A445, 1.301.50 B. The Sample solution in the Assay exhibits a major peak for dactinomycin, the retention time of which corresponds to that exhibited by the Standard solution. ASSAY PROCEDURE [NOTEIn this procedure, use a freshly prepared Standard solution and Sample solution, protected from light.] Mobile phase: Acetonitrile, 0.04 M sodium acetate, and 0.07 M acetic acid (46:25:25) Standard solution: 1.2 mg/mL of USP Dactinomycin RS in Mobile phase Sample solution: 1.2 mg/mL of Dactinomycin in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution (3 replicate injections) [NOTEThe retention time for dactinomycin is 25 min.] Suitability requirements Relative standard deviation: NMT 1.0%, dactinomycin (3 replicate injections)
Dactinomycin for Injection

(Comment on this Monograph)id=m21900=Dactinomycin for Injection=D-Monos.pdf) DEFINITION Dactinomycin for Injection is a sterile mixture of Dactinomycin and Mannitol. It contains NLT 90.0% and NMT 120.0% of the labeled amount of C62H86N12O16, the labeled amount being 0.5 mg in each container. [CAUTIONGreat care should be taken to prevent inhaling particles of Dactinomycin and exposing the skin to it.] IDENTIFICATION A. PROCEDURE Sample solution: 25 g/mL of dactinomycin in methanol Standard solution: 25 g/mL of USP Dactinomycin RS in methanol Acceptance criteria: The UV absorption spectrum of the Sample solution exhibits maxima and minima at the same wavelengths as the Standard solution, concomitantly measured. Ratio: A240/A445, 1.301.50 B. The retention time of the major peak for dactinomycin of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay; and both chromatograms compare qualitatively. ASSAY PROCEDURE [NOTEUse freshly prepared Standard solution and Sample solution, protected from light.]
Dactinomycin / Official Monographs

Mobile phase: Acetonitrile and water (3:2) [NOTEThe acetonitrile concentration may be varied to provide appropriate Chromatographic system performance and to provide a suitable elution time.] Standard solution: 250 g/mL of dactinomycin from USP Dactinomycin RS in Mobile phase Sample solution: 250 g/mL of dactinomycin from Dactinomycin for Injection diluted with Mobile phase. Filter, if necessary, to obtain a clear solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2.5 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe retention time for dactinomycin is 6 min.] Suitability requirements Column efficiency: NLT 1200 theoretical plates Tailing factor: NMT 2 Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C62H86N12O16 taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Dactinomycin RS in the Standard solution (g/mL) CU = nominal concentration of dactinomycin in the Sample solution (g/mL) Acceptance criteria: 90.0%120.0% rU rS CS
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Danazol
(Comment on this Monograph)id=m21950=Danazol=DMonos.pdf)
337.46 C22H27NO2 Pregna-2,4-dien-20-yno[2,3-d]isoxazol-17-ol, (17)-; 17-Pregna-2,4-dien-20-yno[2,3-d]isoxazol-17-ol [17230-88-5]. DEFINITION Danazol contains NLT 97.0% and NMT 102.0% of C22H27NO2, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: Prepare as directed in the Assay. ASSAY PROCEDURE Standard solution: 20 g/mL of previously dried Danazol in alcohol Sample solution: 20 g/mL of USP Danazol RS in alcohol Spectrometric conditions Mode: UV Analytical wavelength: 285 nm Cell: 1 cm Blank: Alcohol Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H27NO2 in the portion of Danazol taken: Result = (AU/AS) (CS/CU) 100 = absorbances of the Sample solution = absorbances of the Standard solution = concentration of USP Danazol RS in the Standard solution (g/mL) CU = concentration for the Sample solution (g/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Organic Impurities PROCEDURE Diluent: Methanol and chloroform (1:9) Standard solutions: 1 mg/mL of USP Danazol RS in Diluent. Dilute quantitatively with Diluent to obtain Standard solutions having the following compositions:
Percentage (%, for comparison with test specimen) 1.0 0.5
SPECIFIC TESTS PH 791: 5.57.5, in the solution constituted as directed in the labeling. LOSS ON DRYING 731: Dry it in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 4% of its weight. OTHER REQUIREMENTS: It meets the requirements under Injections 1. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 100.0 USP Endotoxin Units/mg of dactinomycin. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration, each container being constituted aseptically by injecting Sterile Water for Injection through the stopper, and the entire contents of all the containers being collected aseptically with the aid of 200 mL of Fluid A before filtering. CONSTITUTED SOLUTION: At the time of use, it meets the requirements for Injections 1, Constituted Solutions. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in light-resistant Containers for Sterile Solids as described under Injections 1. LABELING: Label it to include the statement Protect from light. USP REFERENCE STANDARDS 11 USP Dactinomycin RS USP Endotoxin RS
AU AS CS
Standard solution A B
Dilution (1 in 2) (1 in 4)
Concentration (g RS/mL) 500 250
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Percentage (%, for comparison with test specimen) 0.2 0.1
Official Monographs / Danazol 5

Sample solution: Weigh the contents of NLT 20 Capsules. Transfer a portion of the powder, nominally equivalent to 100 mg of danazol, to a 100-mL volumetric flask. Add 50 mL of Mobile phase, and shake by mechanical means for 10 min. Dilute with Mobile phase to volume and filter, discarding the first 5 mL of the filtrate. Pipet 5 mL of the filtrate into a 25-mL volumetric flask, dilute with Mobile phase to volume. Pass a portion of this solution through a 0.45-m porosity filter, discarding the first 5 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 270 nm Column: 3.9-mm 15-cm; 4-m packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0%, danazol Analysis Samples: Standard solution and Sample solution Calculate the percentage of danazol in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentrationof USP Danazol RS in the Standard solution (mg/mL) = nominal concentration for the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.75% sodium lauryl sulfate solution; 900 mL Apparatus 2: 75 rpm Time: 30 min Sample solutions: Sample per Dissolution 711. Dilute with Medium as needed. Standard stock solution: 1 mg/mL of USP Danazol RS in isopropyl alcohol Standard solution: 0.02 mg/mL Standard stock solution and dissolution Medium Analysis Samples: Standard solution and Sample solution Remove an aliquot from the solution under test at a point midway between the stirring shaft and the wall of the vessel and approximately midway in depth. Tolerances: NLT 75% (Q) of the labeled amount of C22H27NO2 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Danazol RS
Standard solution C D
Dilution (1 in 10) (1 in 20)
Concentration (g RS/mL) 100 50
Sample solution: 50 mg/mL of Danazol in Diluent Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Cyclohexane and ethyl acetate (7:3) Analysis Samples: Standard solutions and Sample solution Examine the plate under short-wavelength UV light. Expose the plate to iodine vapors for 5 min. Compare the intensities of any secondary spots observed in the chromatogram of the Sample solution with those of the principal spots in the chromatograms of the Standard solutions. Acceptance criteria: The sum of the intensities of secondary spots obtained from the Sample solution corresponds to NMT 1.0% of related compounds, with no single impurity corresponding to more than 0.5%. SPECIFIC TESTS SPECIFIC ROTATION 781S: + 21 to + 27 Sample solution: 10 mg/mL, in chloroform LOSS ON DRYING 731: Dry it at a pressure not exceeding 5 mm of mercury at 60 to constant weight: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Danazol RS
Danazol Capsules
(Comment on this Monograph)id=m21960=Danazol Capsules=D-Monos.pdf) DEFINITION Danazol Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of C22H27NO2. IDENTIFICATION PROCEDURE Sample solution: 1 mg/mL of danazol from Capsules in chloroform Analysis: Shake the Sample solution. Evaporate the filtrate on a steam bath with the aid of a stream of nitrogen to dryness. Acceptance criteria: The IR absorption spectrum of a potassium bromide dispersion of the residue, previously dried, exhibits maxima at the same wavelengths as that of a similar preparation of USP Danazol RS. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, and water (4:3:3) Standard solution: 0.2 mg/mL of USP Danazol RS in Mobile phase
Dantrolene / Official Monographs
USP 32
Standard stock solution: 50 mg of USP Dantrolene RS in a 50-mL volumetric flask, and dissolve in 2.5 mL of dimethylformamide. Add 2.5 mL of glacial acetic acid, and dilute with acetone to volume to obtain 1.0 mg/mL of dantrolene. Standard solution: 0.1 mg/mL of dantrolene in Diluent, from Standard stock solution Sample stock solution: Dantrolene Sodium in a 100-mL volumetric flask, and dissolve in 5 mL of dimethylformamide. Add 5 mL of glacial acetic acid, and dilute with acetone to volume to obtain 1.25 mg/mL of dantrolene sodium. Sample solution: 0.125 mg/mL of dantrolene sodium in Diluent, from Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 365 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEFor the purpose of identification, the approximate relative retention times for dantrolene related compound B, dantrolene related compound C, and dantrolene are 0.68, 1.24, and 1.0 respectively, Standard solution.] Suitability requirements Resolution: NLT 8 between dantrolene and dantrolene related compound C, System suitabilitiy solution Tailing factor: NMT 1.5, Standard solution Relative standard deviation: NMT 1.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H10N4O5 in the portion of Dantrolene Sodium taken: Result = (rU/rS) (CS/CU) 100 = peak response of dantrolene from the Sample solution rS = peak response of dantrolene from the Standard solution = concentration of USP Dantrolene RS in the CS Standard solution (mg/mL) CU = concentration of dantrolene in the Sample solution (mg/mL) Acceptance criteria: 90.0%96.0% rU IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1: LIMIT OF DANTROLENE RELATED COMPOUND A Mobile phase: Acetonitrile and water (80:20) Standard stock solution: 17.5 g/mL of USP Dantrolene Related Compound A RS and 50 g/mL of USP Dantrolene Sodium RS in dimethylformamide Standard solution: 0.35 g/mL of dantrolene related compound A and 1 g/mL of dantrolene sodium in acetonitrile Sample stock solution: Prepare as directed for the Sample stock solution in the Assay. Sample solution: 0.175 mg/mL of dantrolene sodium in acetonitrile, from Sample stock solution Chromatographic system (See Chromatography 621, System Suitability.)
Dantrolene Sodium
(Comment on this Monograph)id=m22060=Dantrolene Sodium=D-Monos.pdf)
399.29 C14H9N4NaO5 31/2H2O 2,4-Imidazolidinedione, 1-[[[5-(4-nitrophenyl)-2furanyl]methylene]amino]-, sodium salt, hydrate (2:7); 1-[[5-(p-Nitrophenyl)furfurylidene]amino]hydantoin sodium salt hydrate [24868-20-0]. DEFINITION Dantrolene Sodium contains NLT 90.0% and NMT 96.0% of C14H10N4O5, the free acid form of Dantrolene Sodium, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Sodium 191: Ignite about 200 mg: the residue meets the requirements of the flame test for sodium. ASSAY PROCEDURE Buffer: 3.85 mg/mL of ammonium acetate, and adjust with glacial acetic acid to a pH of 4.5 0.1 Diluent: Acetonitrile and water (50:50) Solution A: Acetonitrile, Buffer, and water (10:20:70) Solution B: Acetonitrile and Buffer (80:20) Mobile phase: See gradient table below.
Time (min) 0 10 20 25 25.1 35 Solution A (%) 90 60 10 10 90 90 Solution B (%) 10 40 90 90 10 10
System suitability stock solution A: 62.5 mg of USP Dantrolene Sodium RS in a 50-mL volumetric flask, and dissolve in 2.5 mL of dimethylformamide. Add 2.5 mL of glacial acetic acid, and dilute with acetone to volume to obtain 1.25 mg/mL of dantrolene sodium. System suitability stock solution B: 6.3 mg each of USP Dantrolene Related Compound B RS and USP Dantrolene Related Compound C RS in a 50-mL volumetric flask, and dissolve in 2.5 mL of dimethylformamide. Add 2.5 mL of glacial acetic acid, and dilute with acetone to volume to obtain 0.125 mg/mL each of dantrolene related compound B and dantrolene related compound C. System suitability solution: 0.125 mg/mL of dantrolene sodium in Diluent, from System suitability stock solution A; 2.5 g/mL each of dantrolene related compound B and dantrolene related compound C in Diluent, from System suitability stock solution B
USP 32
Mode: LC Detector: UV 365 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements: Tailing factor: NMT 1.5, dantrolene related compound A Relative standard deviation: NMT 5%, dantrolene related compound A [NOTEThe dantrolene peak elutes at void volume at approximately 1.5 min.] Analysis Samples: Standard solution and Sample solution Calculate the percentage of dantrolene related compound A in the portion of Dantrolene Sodium taken: Result = (rU/rS) (CS/CU) 100 = peak response of dantrolene related compound A from the Sample solution = peak response of dantrolene related compound A rS from the Standard solution CS = concentration of dantrolene related comnpound A in the Standard solution (mg/mL) CU = concentration of Dantrolene Sodium in the Sample solution (mg/mL) Acceptance criteria: NMT 0.15% PROCEDURE 2 Diluent, Mobile phase, System suitability stock solution B, and Chromatographic system: Prepare as directed in the Assay. Standard solution: Dilute quantitatively System suitability stock solution B with Diluent to obtain 0.25 g/mL each of dantrolene related compound B and dantrolene related compound C. Sample solution: Use the Sample solution from the Assay Calculate the percentage of each of the relevant dantrolene related compounds in the portion of Dantrolene Sodium taken: rU Result = (rU/rS) (CS/CU) 100 = individual peak response for dantrolene related compound B or dantrolene related compound C from the Sample solution = response of the corresponding peak of the rS Standard solution = concentration of dantrolene related compound CS B or dantrolene related compound C in the Standard solution (mg/mL) = concentration of Dantrolene Sodium in the CU Sample solution (mg/mL) Acceptance criteria: NMT 0.50% for dantrolene related compound B, and NMT 0.30% for dantrolene related compound C rU SPECIFIC TESTS WATER, Method Ia 921: 14.5%17.0%
Official Monographs / Dantrolene 7
Dantrolene Sodium Capsules

(Comment on this Monograph)id=m22070=Dantrolene Sodium Capsules=D-Monos.pdf) DEFINITION Dantrolene Sodium Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of dantrolene sodium (C14H9N4NaO5 31/2H2O). IDENTIFICATION A. INFRARED ABSORPTION 197K B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Diluent: Acetonitrile and water (70:30) Buffer solution: 3.3 mg/mL of ammonium acetate Solution A: Acetonitrile, glacial acetic acid, and Buffer solution (76:7:120) Solution B: Acetonitrile and water (70:30) Mobile phase: See gradient table below.
Time (min) 0 8 8.1 13 13.1 20 Solution A (%) 100 100 0 0 100 100 Solution B (%) 0 0 100 100 0 0
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at room temperature. USP REFERENCE STANDARDS 11 USP Dantrolene RS USP Dantrolene Related Compound A RS USP Dantrolene Related Compound B RS USP Dantrolene Related Compound C RS USP Dantrolene Sodium RS
System suitability solution: Use the Standard solution, prepared as directed under Organic Impurities. Standard stock solution: 40 mg of USP Dantrolene RS in a 50-mL volumetric flask, and dissolve in 2.5 mL of dimethylformamide. Add 2.5 mL of glacial acetic acid, and dilute with acetone to volume. The final concentration is about 0.8 mg/mL of dantrolene. Standard solution: 0.08 mg/mL of dantrolene in Diluent, made from Standard stock solution Sample stock solution: Mix the combined contents of NLT 20 Capsules, and transfer a portion, equivalent to the average weight of one Capsule, to a 50-mL volumetric flask. Add 10 mL of dimethylformamide, and sonicate for 15 min to dissolve. Add 5 mL of glacial acetic acid, and dilute with acetone to volume. Sample solution: 0.1 mg/mL of dantrolene sodium in Diluent, made from Sample stock solution. Pass through a 0.45-m nylon filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 365 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Samples: System suitability solution and Standard solution [NOTEFor the purpose of peak identification, the approximate relative retention times for dantrolene related compound B and dantrolene are about 0.68 and 1.0, respectively.]

Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.0% for dantrolene Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H9N4NaO5 31/2H2O in the portion of Capsules taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response of dantrolene from the Sample solution = peak response of dantrolene from the Standard rS solution = concentration of dantrolene in the Standard CS solution (mg/mL) CU = nominal concentration of dantrolene in the Sample solution (mg/mL) = molecular weight of dantrolene sodium, 399.29 Mr1 = molecular weight of dantrolene, 314.25 Mr2 Acceptance criteria: 90.0%110.0% rU
USP 32
absorbance of Standard solution 2 is between 0.3 and 0.5; and the ratio of the absorbance of Standard solution 1 to that of Standard solution 3 is 4.00 0.10. Analysis Samples: Standard solution and Sample solution Determine the amount of C14H9N4NaO5 31/2H2O dissolved by measuring the absorbance of the Sample solution at the wavelength of maximum absorbance at about 395 nm in comparison with the appropriate Standard solution, using a 0.1-cm cell and Medium as the blank. All absorbance values are obtained on solutions within 2 h of their preparation. Calculate the percentage of C14H9N4NaO5 31/2H2O dissolved: Result = (AU CS V 100)/(AS F L) = absorbance of the Sample solution = absorbance of the Standard solution = concentration of dantrolene in the Standard solution (mg/mL) V = volume of Medium (mL), 900 F = correction for water of hydration and sodium contained in the dantrolene sodium monohydrate form of the drug, assuming that the bulk contains 15% of water and 6.84% of sodium, 0.79186 L = capsule label claim (mg) Tolerances: NLT 75% (Q) of the labeled amount of C14H9N4NaO5 31/2H2O is dissolved. AU AS CS IMPURITIES Organic Impurities PROCEDURE Diluent, Solution A, Solution B, Mobile phase, and Chromatographic system: Proceed as directed in the Assay. Standard stock solution: 5 mg of USP Dantrolene Related Compound B RS dissolved in 2.5 mL of dimethylformamide in a 50-mL volumetric flask. Add 2.5 mL of glacial acetic acid, and dilute with acetone to volume. The final concentration is about 0.1 mg/mL. Standard solution: 0.0005 mg/mL of dantrolene related compound B in Diluent, from Standard stock solution Sample solution: Use the Sample solution as prepared in the Assay. Analysis Samples: Standard solution and Sample solution Calculate the percentage of dantrolene related compound B in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response of dantrolene related compound B from the Sample solution = corresponding peak response from the Standard rS solution = concentration of dantrolene related compound CS B in the Standard solution (mg/mL) = nominal concentration of dantrolene sodium in CU the Sample solution (mg/mL) Acceptance criteria: NMT 2% UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements rU ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Dantrolene RS USP Dantrolene Related Compound B RS USP Dantrolene Sodium RS
PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.5% methylbenzethonium chloride in water, pH 6.8; 900 mL, deaerated Apparatus 1: 100 rpm Time: 40 min Determine the amount of C14H9N4NaO5 31/2H2O dissolved by employing the following method. Standard solution 1 (for Capsules labeled to contain 100 mg): 25 mg of USP Dantrolene RS in a 250-mL volumetric flask. Dissolve in 5.0 mL of dimethylformamide. Add 200 mL of Medium and 10.0 mL of 0.1 N potassium hydroxide. Dilute with Medium to volume. Pass through a 0.45-m polytetrafluoroethylene (PTFE) filter, previously wetted with a few drops of isopropyl alcohol, discarding the first 5 mL. Standard solution 2 (for Capsules labeled to contain 50 mg): 25.0 mL of Standard solution 1 in a 50-mL volumetric flask containing 0.5 mL of 0.1 N potassium hydroxide. Dilute with Medium to volume. Pass through a 0.45-m PTFE filter, previously wetted with a few drops of isopropyl alcohol, discarding the first 5 mL. Standard solution 3 (for Capsules labeled to contain 25 mg): 25.0 mL of Standard solution 1 in a 100-mL volumetric flask containing 1.0 mL of 0.1 N potassium hydroxide. Dilute with Medium to volume. Pass through a 0.45-m PTFE filter, previously wetted with a few drops of isopropyl alcohol, discarding the first 5 mL. Sample solution: Withdraw 10 mL of the solution under test. Pass through a 0.45-m PTFE filter, previously wetted with a few drops of isopropyl alcohol. Discard the first 5 mL. Collect the filtered solution in a tube that contains 1 drop of 1 N potassium hydroxide. Spectrometric conditions Mode: UV Analytical wavelength: 395 nm Cell: 0.1 cm Blank: Medium System suitability Samples: Medium, Standard solution 1, Standard solution 2 and Standard solution 3 [NOTEAll absorbance values should be obtained on solutions within 2 h of their preparation.] Using a 0.1-cm cell, measure the absorbance of Medium, using water as the blank, and measure the absorbance of each of the three Standard solutions using Medium as the blank, at the wavelength of maximum absorbance at about 395 nm. The system is considered suitable for use if the following criteria are met: the absorbance of Medium is less than 10% of the absorbance of Standard solution 1; the
USP 32
Official Monographs / Dantrolene 9

Mode: LC Detector: UV 365 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.5% of dantrolene Analysis Samples: Standard solution and Sample solution Calculate the percentage of C14H9N4NaO5 31/2H2O in the portion of Dantrolene Sodium for Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Dantrolene RS in the Standard solution (mg/mL) = nominal concentration of dantrolene sodium CU hydrate in the Sample solution (mg/mL) = molecular weight of dantrolene sodium hydrate, Mr1 399.29 = molecular weight of dantrolene, 314.25 Mr2 Acceptance criteria: 90.0%110.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements
Dantrolene Sodium for Injection

(Comment on this Monograph)id=m22080=Dantrolene Sodium for Injection=D-Monos.pdf) DEFINITION Dantrolene Sodium for Injection is a sterile, non-pyrogenic, lyophilized formulation containing Dantrolene Sodium, and one or more suitable buffering or sequestering agents. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C14H9N4NaO5 31/2H2O. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: To 0.5 g of Dantrolene Sodium for Injection, add 10 mL of 0.1 N hydrochloric acid and 10 mL of ethyl acetate. Allow the phases to separate, and transfer the upper ethyl acetate phase to a suitable glass container. Evaporate the solvent, dry the residue at 105 for 10 min, and use the residue. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: Dissolve 3.3 mg of ammonium acetate in 1 L of water, and adjust with acetic acid to a pH of 4.5 0.1. Solution A: Acetonitrile, Buffer, and glacial acetic acid (80:120:7) Solution B: Acetonitrile and water (70:30) Mobile phase: See the gradient table below.
Time (min) 0 8 8.1 13 13.1 20 Solution A (%) 100 100 0 0 100 100 Solution B (%) 0 0 100 100 0 0
Diluent: Acetonitrile and water (60:40) Standard stock solution: 40 mg of USP Dantrolene RS in a 50-mL volumetric flask. Dissolve in 2.5 mL of dimethylformamide. Add 2.5 mL of glacial acetic acid, and dilute with acetone to volume. The final concentration is about 0.8 mg/mL. Standard solution: Nominally 0.08 mg/mL of dantrolene, from Standard stock solution, diluted with Diluent Sample solution: Using 70 mL of water for each vial, transfer the entire contents of the required number of vials to a suitable flask necessary to obtain a solution having a known concentration of about 0.1 mg/mL of dantrolene sodium hydrate. Sonicate for 25 min to dissolve the sample. Dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.)
IMPURITIES Organic Impurities PROCEDURE Mobile phase and Diluent: Proceed as directed in the Assay. Standard stock solution: 10 mg of USP Dantrolene Related Compound B RS in a 50-mL volumetric flask. Dissolve in 2.5 mL of dimethylformamide. Add 2.5 mL of glacial acetic acid, and dilute with acetone to volume. The final concentration is about 0.2 mg/mL. Standard solution: 0.002 mg/mL of dantrolene related compound B in Diluent from Standard stock solution Sample solution: Use Sample solution from the Assay. Chromatographic system: Proceed as directed in the Assay. System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 5.0% of dantrolene related compound B Analysis Samples: Standard solution and Sample solution Calculate the percentage of dantrolene related compound B in the portion of Dantrolene Sodium for Injection taken: Result = (rU/rS) (CS/CU) 100 rU = peak response of the Sample solution
10

= corresponding peak response of the Standard solution = concentration of dantrolene related compound B CS in the Standard solution (mg/mL) = nominal concentration of dantrolene sodium CU hydrate in the Sample solution (mg/mL) Acceptance criteria: NMT 8% rS
USP 32
Suitability requirements Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H12N2O2S in the portion of Dapsone taken: Result = (rU/rS) (CS/CU) 100 = peak responses of the Sample solution = peak responses of the Standard solution = concentration of USP Dapsone RS in the Standard solution (g/mL) CU = concentration of Dapsone in the Sample solution (g/mL) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% SELENIUM 291: 0.003%, a 100-mg sample mixed with 100 mg of magnesium oxide being used Organic Impurities PROCEDURE Standard solution A: 12.5 mg/mL of USP Dapsone RS in methanol Standard solution B: 125 g/mL of USP Dapsone RS in methanol from Standard solution A Standard solution C: 62.5 g/mL of USP Dapsone RS in methanol from Standard solution A Sample solution: 12.5 mg/mL of Dapsone in methanol Chromatographic system: (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 150- to 200-m layer of chromatographic silica gel Application volume: 4 L Developing solvent system: Acetone, chloroform, nbutyl alcohol, and formic acid (15:60:15:10) [NOTEPrepare the solvent system fresh daily. Equilibrate the chromatographic chamber with the solvent system for 30 min prior to developing the chromatographic plate.] Spray reagent: 0.1% (w/v) solution of 4dimethylaminocinnamaldehyde in glacial acetic acid and water (1:1) Analysis Samples: Standard solutions and Sample solution Position the plate in a chromatographic chamber, and develop the chromatograms in the developing solvent system until the solvent front has moved about threefourths of the length of the plate. Remove the plate from the developing chamber and air-dry. Spray the plate lightly with a Spray reagent. Examine the spots that are developed immediately, and compare the intensities of any secondary spots observed in the Sample solution with those of the principal spots of the Standard solutions. Acceptance criteria: No secondary spot from the chromatogram of the Sample solution is larger or more intense than the principal spot of Standard solution C (0.5%), and the sum of the intensities of all the secondary spots of the Sample solution corresponds to NMT 1.0%. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 175181 LOSS ON DRYING 731: Dry at 105 for 3 h: it loses NMT 1.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. rU rS CS
SPECIFIC TESTS PH 791: 8.811.0 Dissolve the contents of 1 vial in 60 mL of USP Water for Injection. WATER DETERMINATION, Method Ia 921: NMT 3% OTHER REQUIREMENTS: It meets the requirements under Injections 1. STERILITY TESTS 71: Meets the requirements BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.5 USP Endotoxin Unit/mg of dantrolene sodium. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. Store at controlled room temperature, protected from light. USP REFERENCE STANDARDS 11 USP Dantrolene RS USP Dantrolene Related Compound B RS USP Endotoxin RS
Dapsone
(Comment on this Monograph)id=m22120=Dapsone=DMonos.pdf)
C12H12N2O2S Benzenamine, 4,4-sulfonylbis-; 4,4-Sulfonyldianiline [80-08-0]. DEFINITION Dapsone contains NLT 98.0% and NMT 102.0% of C12H12N2O2S, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 5 g/mL in methanol
248.30
ASSAY PROCEDURE Mobile phase: Transfer 100 mL of isopropyl alcohol, 100 mL of acetonitrile, and 100 mL of ethyl acetate to a 1000mL volumetric flask. Add hexane to volume without mixing, then mix, and allow the mixture to cool to room temperature. Standard solution: 25 g/mL of USP Dapsone RS in Mobile phase Sample solution: 25 g/mL of Dapsone Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; 10-m diameter packing L3 Injection size: 10 L System suitability Sample: Standard solution (chromatograph a sufficient number)
USP 32
USP REFERENCE STANDARDS 11 USP Dapsone RS rU rS CS
Official Monographs / Dapsone 11

= peak responses of the Sample solution = peak responses of the Standard solution = concentration of USP Dapsone RS in the Standard solution (g/mL) CU = nominal concentration of dapsone in the Sample solution (g/mL) Acceptance criteria: 92.5%107.5% PERFORMANCE TESTS DISSOLUTION 711 Medium: Dilute hydrochloric acid (2 in 100); 1000 mL Apparatus 1: 100 rpm Time: 60 min Sample solution: Sample per Dissolution 711. Withdraw and filter a portion of the Sample solution. Transfer an accurately measured portion of the filtrate estimated to contain 0.2 mg of dapsone to a 25-mL volumetric flask, add 5 mL of 1 N sodium hydroxide, and dilute with water to volume. Standard solution: USP Dapsone RS of a known concentration in Medium Spectrometric conditions Mode: UV Analytical wavelength: 290 nm Tolerances: NLT 75% (Q) of the labeled amount of C12H12N2O2S UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Analysis for content uniformity Sample solution: To 1 Tablet in a 100-mL volumetric flask, add 2.0 mL of water, and allow to stand for 30 min, swirling occasionally. Add 70 mL of methanol, and place the flask in an ultrasonic bath until the specimen is completely dispersed. Add methanol to volume, and centrifuge a portion of the mixture. Quantitatively dilute a measured volume of the clear supernatant with methanol to obtain a solution having a concentration of 8 g/mL of dapsone. Standard solution: 8 g/mL of USP Dapsone RS in methanol Spectrometric conditions Mode: UV Analytical wavelength: 296 nm Cell: 1 cm Blank: Methanol Analysis Samples: Standard solution and Sample solution Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbances of the solution from the Tablet = absorbances of the Standard solution = concentration of USP Dapsone RS in the Standard solution (g/mL) = nominal concentration of dapsone in the Sample solution (g/mL)
Dapsone Tablets
(Comment on this Monograph)id=m22150=Dapsone Tablets=DMonos.pdf) DEFINITION Dapsone Tablets contain NLT 92.5% and NMT 107.5% of the labeled amount of C12H12N2O2S. IDENTIFICATION A. PROCEDURE Sample solution: Transfer a quantity of finely powdered Tablets, equivalent to 100 mg of dapsone, to a suitable container, add 5 mL of acetone, shake for 5 min, filter, and evaporate the filtrate to dryness. Dry this residue at 105 for 1 h: the residue so obtained meets the requirements of the test for Infrared Absorption 197K. B. PROCEDURE Sample solution: Triturate a quantity of finely powdered Tablets equivalent to 100 mg of dapsone with 50 mL of methanol, and filter. Dilute a portion of the filtrate with methanol to make approximately a 1 in 200,000 solution: this solution meets the requirements of the test for Ultraviolet Absorption 197U. ASSAY PROCEDURE Mobile phase: Transfer 100 mL of isopropyl alcohol, 100 mL of acetonitrile, and 100 mL of ethyl acetate to a 1000mL volumetric flask. Add hexane to volume without mixing, then mix, and allow the mixture to cool to room temperature. Standard solution: Dissolve USP Dapsone RS in Mobile phase to obtain a solution having a known concentration of 250 g/mL. Pipet 5 mL of this solution into a 50-mL volumetric flask, dilute with Mobile phase to volume, and mix to obtain a solution having a known concentration of 25 g/mL. Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder nominally equivalent to 50 mg of dapsone to a 200-mL volumetric flask. Add 150 mL of methanol, and place the flask in an ultrasonic bath at a temperature of 35 for 15 min, with occasional shaking. Allow to cool to room temperature, and add methanol to volume. Centrifuge a portion of the mixture until clear. Transfer 5.0 mL of the clear supernatant to a 50-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; 10-m diameter packing L3 Injection size: 10 L System suitability Sample: Standard solution (chromatograph a sufficient number) Suitability requirements Relative standard deviation: NMT 2% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C12H12N2O2S in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. USP REFERENCE STANDARDS 11 USP Dapsone RS
12
Daunorubicin / Official Monographs

rU
USP 32
= peak response of daunorubicin from the Sample solution rS = peak response of daunorubicin from the Standard solution CS = concentration of daunorubicin in the Standard solution (g/mL) W = sample weight of Daunorubicin Hydrochloride taken (mg) V = volume of the Sample solution Acceptance criteria: 8421030 g SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 4.56.5, in a solution containing 5 mg/mL WATER DETERMINATION, Method I 921: NMT 3.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light and excessive heat. USP REFERENCE STANDARDS 11 USP Daunorubicin Hydrochloride RS
Daunorubicin Hydrochloride
(Comment on this Monograph)id=m22200=Daunorubicin Hydrochloride=D-Monos.pdf)
C27H29NO10 HCl 563.98 5,12-Naphthacenedione, 8-acetyl-10-[(3-amino-2,3,6-trideoxy-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11trihydroxy-1-methoxy-, (8S-cis)-, hydrochloride; (1S,3S)-3-Acetyl-1,2,3,4,6,11-hexahydro-3,5,12-trihydroxy-10methoxy-6,11-dioxo-1-naphthacenyl 3-amino-2,3,6-trideoxy-L-lyxo-hexopyranoside hydrochloride [23541-50-6]. DEFINITION Daunorubicin Hydrochloride has a potency equivalent to NLT 842 g and NMT 1030 g of C27H29NO10 per mg. [CAUTIONGreat care should be taken to prevent inhaling particles of Daunorubicin Hydrochloride and exposing the skin to it.] IDENTIFICATION A. The IR absorption spectrum of a potassium bromide dispersion of it exhibits maxima only at the same wavelengths as that of a similar preparation of USP Daunorubicin Hydrochloride RS. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (19:31), adjust with phosphoric acid to a pH of 2.2 0.2. The acetonitrile concentration may be varied to meet system suitability requirements and to provide a suitable elution time for daunorubicin. Pass the solution through a membrane filter (1-m or finer porosity). Standard solution: 250 g/mL of daunorubicin from USP Daunorubicin Hydrochloride RS in Mobile phase System suitability solution: 250 g/mL of doxorubicin hydrochloride in the Standard solution Sample solution: 0.25 mg/mL of Daunorubicin Hydrochloride in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 5 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for doxorubicin and daunorubicin are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 3 between the doxorubicin and the daunorubicin peaks, System suitability solution Relative standard deviation: NMT 2.0% for replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the potency, in g of C27H29NO10 per mg, taken: Result = (rU/rS) (CS/W) V
Daunorubicin Hydrochloride for Injection

(Comment on this Monograph)id=m22210=Daunorubicin Hydrochloride for Injection=D-Monos.pdf) DEFINITION Daunorubicin Hydrochloride for Injection is a sterile mixture of Daunorubicin Hydrochloride and Mannitol. It contains the equivalent of NLT 90.0% and NMT 115.0% of the labeled amount of C27H29NO10. IDENTIFICATION The retention time of the main peak of the Sample solution corresponds to that of the Standard solution, as directed in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (19:31), adjust with phosphoric acid to a pH of 2.2 0.2. The acetonitrile concentration may be varied to meet system suitability requirements and to provide a suitable elution time for daunorubicin. Pass the solution through a membrane filter (1-m or finer porosity). Standard solution: 250 g/mL of daunorubicin from USP Daunorubicin Hydrochloride RS in Mobile phase System suitability solution: 250 g/mL of doxorubicin hydrochloride in the Standard solution Sample solution: Transfer the contents of 1 vial of Daunorubicin Hydrochloride for Injection with the aid of Mobile phase to an appropriate volumetric flask, and dilute with Mobile phase to volume to obtain a solution containing nominally 0.25 mg/mL of daunorubicin. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 5 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for doxorubicin and daunorubicin are about 0.7 and 1.0, respectively.]
USP 32
Suitability requirements Resolution: NLT 3 between the doxorubicin and the daunorubicin peaks, System suitability solution Relative standard deviation: NMT 2.0% for replicate injections, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C27H29NO10 in the vial of Daunorubicin Hydrochloride for Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response of daunorubicin from the Sample solution = peak response of daunorubicin from the rS Standard solution = concentration of USP Daunorubicin RS in the CS Standard solution (g/mL) = nominal concentration of daunorubicin in the CU Sample solution (g/mL) Acceptance criteria: 90.0%115.0% rU SPECIFIC TESTS PH 791: 4.56.5, in the solution constituted as directed in the labeling WATER DETERMINATION, Method I 921: NMT 3.0%, the Sample being prepared as directed for a hygroscopic specimen CONSTITUTED SOLUTION: At the time of use, it meets the requirements under Injections 1, Constituted Solutions. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 4.3 USP Endotoxin Units/mg of daunorubicin. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in light-resistant Containers for Sterile Solids as described under Injections 1. USP REFERENCE STANDARDS 11 USP Daunorubicin Hydrochloride RS USP Endotoxin RS
Official Monographs / Decoquinate 13

volume. Promptly transfer 10.0 mL of this solution to a second 100-mL volumetric flask, and dilute with dehydrated alcohol to volume. Transfer 10.0 mL of this solution to a third 100-mL volumetric flask, add 10 mL of 0.1 N hydrochloric acid, and dilute with dehydrated alcohol to volume. Acceptance criteria: The UV absorption spectrum of this solution exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Decoquinate RS, concurrently measured. Absorptivities do not differ by more than 2.5% calculated on the dried basis, at the wavelength of maximum absorption at 265 nm. ASSAY PROCEDURE Sample: 1000 mg of Decoquinate Analysis: Dissolve in 100 mL of glacial acetic acid with the aid of gentle heat. Allow to cool, add 1 drop of crystal violet TS, and titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 41.76 mg of C24H35NO5. Acceptance criteria: 99.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE: ORDINARY IMPURITIES 466 Sample solution: Chloroform, prepared with the aid of heat Standard solution: Chloroform, using dilutions of the Sample solution Eluant: A mixture of chloroform, dehydrated alcohol, and anhydrous formic acid (85:10:5) Visualization: 1 Tolerances: No impurity exceeds 1%, and the total does not exceed 2%. SPECIFIC TESTS LOSS ON DRYING 731: Dry at 105 to constant weight: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Decoquinate RS
Decoquinate
(Comment on this Monograph)id=m22310=Decoquinate=DMonos.pdf)
Decoquinate Premix
417.54 C24H35NO5 3-Quinolinecarboxylic acid, 6-(decyloxy)-7-ethoxy-4-hydroxy-, ethyl ester; Ethyl 6-(decyloxy)-7-ethoxy-4-hydroxy-3-quinolinecarboxylate [18507-89-6]. DEFINITION Decoquinate contains NLT 99.0% and NMT 101.0% of C24H35NO5, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solution: Transfer 40 mg of Decoquinate to a 100mL volumetric flask, add 10 mL of hot chloroform, swirl to dissolve, and while still warm, add 60 mL of dehydrated alcohol. Allow to cool, and dilute with dehydrated alcohol to (Comment on this Monograph)id=m22315=Decoquinate Premix=D-Monos.pdf) DEFINITION Decoquinate Premix contains NLT 90.0% and NMT 110.0% of the labeled amount of C24H35NO5, the labeled amount being 110 g/100 g of Premix. IDENTIFICATION PROCEDURE Standard solution: 2.5 mg/mL of USP Decoquinate RS in chloroform Sample solution: To a weighed quantity of sample, equivalent to 100 mg of decoquinate, add 40 mL of chloroform, and heat under a reflux condenser on a water bath for 20 min, cool, and filter. Use the filtrate as the Sample solution. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
14
Decoquinate / Official Monographs
USP 32
Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L (apply to the starting line of chromatographic plate) Developing solvent system: Chloroform and alcohol (7:3) Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram in Developing solvent system until the solvent front has moved about three-fourths of the length of the plate. Locate the spots under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Standard solution: Transfer 50 mg of USP Decoquinate RS to a 100-mL volumetric flask, add 10 mL of hot chloroform, swirl to dissolve, and while still warm, slowly add 70 mL of dehydrated alcohol. Allow to cool, and dilute with dehydrated alcohol to volume. Immediately transfer 10.0 mL of this solution to a second 100-mL volumetric flask, and dilute with dehydrated alcohol to volume. Transfer 10.0 mL of this solution to a third 100-mL volumetric flask, add 10 mL of 0.1 N hydrochloric acid, and dilute with dehydrated alcohol to volume. This contains 0.005 mg/mL of USP Decoquinate RS. Sample solution: Extract a quantity of Premix, nominally equivalent to 200 mg of decoquinate, with 50 mL of chloroform in a small continuous extraction apparatus for 8 reflux cycles. Transfer the extract to a 100-mL volumetric flask with the aid of chloroform, cool, and dilute with chloroform to volume. Transfer 5.0 mL of this solution to a second 100-mL volumetric flask, and dilute with dehydrated alcohol to volume. Transfer 5.0 mL of this solution to a third 100-mL volumetric flask, add 10 mL of 0.1 N hydrochloric acid, and dilute with dehydrated alcohol to volume. Spectrometric conditions Mode: UV Analytical wavelength: 265 nm Blank: Dehydrated alcohol, 0.1 N hydrochloric acid, and chloroform (90:10:0.25) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C24H35NO5 in the portion of Premix taken: Result = (AU/AS) (CS/CU) 4000 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Decoquinate RS in the Standard solution (mg/mL) = nominal concentration of decoquinate in the CU Sample solution Acceptance criteria: 90.0%110.0% of the labeled amount of C24H35NO5, the labeled amount being 110 g/100 g of Premix AU AS CS ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Decoquinate RS
Deferoxamine Mesylate
(Comment on this Monograph)id=m22400=Deferoxamine Mesylate=D-Monos.pdf)
656.79 C25H48N6O8 CH4O3S Butanediamide, N-[5-[[4-[[5(acetylhydroxyamino)pentyl]amino]-1,4dioxobutyl]hydroxyamino]pentyl]-N-(5-aminopentyl)-Nhydroxy-, monomethanesulfonate; N-[5-[3-[(5-Aminopentyl)hydroxycarbamoyl]propionamido] pentyl]-3-[[5-(N-hydroxyacetamido)pentyl]carbamoyl] propionohydroxamic acid monomethanesulfonate (salt) [138-14-7]. DEFINITION Deferoxamine Mesylate contains NLT 98.0% and NMT 102.0% of C25H48N6O8 CH4O3S, calculated on the anhydrous basis. IDENTIFICATION PROCEDURE Sample: 5 mg Analysis: Dissolve Sample in 5 mL of water, add 2 mL of tribasic sodium phosphate solution (1 in 200), then add 10 drops of -naphthoquinone-4-sodium sulfonate solution (1 in 40). Acceptance criteria: A blackish brown color is produced. ASSAY PROCEDURE Solution A: Dissolve 6.7 g of ferric chloride in dilute hydrochloric acid (1 in 100) in a 100-mL volumetric flask. Add dilute hydrochloric acid (1 in 100) to volume. Standard solution: 1 mg/mL of USP Deferoxamine Mesylate RS Sample solution: 1 mg/mL of Deferoxamine Mesylate Spectrometric conditions Mode: UV-Vis Analytical wavelength: 485 nm Cell: 1 cm Blank: Water Analysis Samples: Standard solution, Sample solution, and Blank Pipet 2 mL each of the Standard solution, the Sample solution, and the Blank into separate 25-mL volumetric flasks. To each flask, add 3 mL of Solution A, and dilute with water to volume. Concomitantly determine the absorbances. Calculate the percentage of C25H48N6O8 CH4O3S in the portion taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance from the Sample solution = absorbance from the Standard solution = concentration of USP Deferoxamine Mesylate RS in the Standard solution (mg/mL) = concentration of Deferoxamine Mesylate in the Sample solution (mg/mL)
USP 32
Official Monographs / Dehydrocholic 15

flask add 3 mL of Solution A, and dilute with water to volume. Concomitantly determine the absorbances. Calculate the percentage of C25H48N6O8 CH4O3S in the vial of Deferoxamine Mesylate for Injection taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Deferoxamine Mesylate RS in the Standard solution (mg/mL) CU = nominal concentration of deferoxamine mesylate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements AU AS CS
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1%, 2.0 g being used CHLORIDE AND SULFATE, Chloride 221: A 1.2-g portion shows no more chloride than corresponds to 0.20 mL of 0.020 N hydrochloric acid (0.012%). CHLORIDE AND SULFATE, Sulfate 221: A 0.5-g portion shows no more sulfate than corresponds to 0.20 mL of 0.020 N sulfuric acid (0.04%). HEAVY METALS, Method II 231: 10 ppm SPECIFIC TESTS PH 791: 4.06.0, in a solution (1 in 100) WATER DETERMINATION, Method I 921: NMT 2.0% STERILITY TESTS 71: Where the label states that Deferoxamine Mesylate is sterile, it meets the requirements. BACTERIAL ENDOTOXINS TEST 85: Where the label states that Deferoxamine Mesylate is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.33 USP Endotoxin Unit/mg of deferoxamine mesylate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Deferoxamine Mesylate RS USP Endotoxin RS
SPECIFIC TESTS PH 791: 4.06.0, in a solution (1 in 100) WATER DETERMINATION, Method I 921: NMT 1.5% OTHER REQUIREMENTS: It meets the requirements under Injections 1. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.33 USP Endotoxin Unit/mg of deferoxamine mesylate. CONSTITUTED SOLUTION: At the time of use, it meets the requirements under Injections 1, Constituted Solutions. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Deferoxamine Mesylate RS USP Endotoxin RS
Deferoxamine Mesylate for Injection

(Comment on this Monograph)id=m22410=Deferoxamine Mesylate for Injection=D-Monos.pdf) DEFINITION Deferoxamine Mesylate for Injection contains NLT 90.0% and NMT 110.0% of the labeled amount of C25H48N6O8 CH4O3S. IDENTIFICATION PROCEDURE Sample: 5 mg of Deferoxamine Mesylate for Injection Analysis: Dissolve Sample in 5 mL of water, add 2 mL of tribasic sodium phosphate solution (1 in 200), then add 10 drops of -naphthoquinone-4-sodium sulfonate solution (1 in 40). Acceptance criteria: A blackish brown color is produced. ASSAY PROCEDURE Solution A: Dissolve 6.7 g of ferric chloride in dilute hydrochloric acid (1 in 100) in a 100-mL volumetric flask. Add dilute hydrochloric acid (1 in 100) to volume. Standard solution: 1 mg/mL of USP Deferoxamine Mesylate RS Sample solution: Constitute the contents of 1 vial in water, and dilute with water to obtain nominally a 1 mg/mL of deferoxamine mesylate. Spectrometric conditions Mode: UV Cell: 1 cm Analytical wavelength: 485 nm Blank: Water Analysis Samples: Standard solution, Sample solution, and Blank Pipet 2 mL each of the Standard solution, Sample solution, and Blank into separate 25-mL volumetric flasks. To each
Dehydrocholic Acid
(Comment on this Monograph)id=m22560=Dehydrocholic Acid=D-Monos.pdf)
C24H34O5 Cholan-24-oic acid, 3,7,12-trioxo-, (5)-; 3,7,12-Trioxo-5-cholan-24-oic acid [81-23-2].
402.52
DEFINITION Dehydrocholic Acid contains NLT 98.5% and NMT 101.0% of C24H34O5, calculated on the dried basis. Dehydrocholic Acid for parenteral use melts between 237 and 242. IDENTIFICATION INFRARED ABSORPTION 197K ASSAY PROCEDURE Sample: 500 mg of Dehydrocholic Acid Analysis: Add 60 mL of neutralized alcohol, and warm on a steam bath until solution is effected. Cool, add phenolphthalein TS and 20 mL of water. Titrate with 0.1 N sodium hydroxide VS, adding 100 mL of water shortly before the endpoint is reached. Each mL of 0.1 N sodium hydroxide is equivalent to 40.25 mg of C24H34O5.
16
Dehydrocholic / Official Monographs

98.5%101.0% Acceptance criteria: 94.0%106.0%
USP 32
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.3% BARIUM: Add to the mixture obtained in the test for Odor on Boiling 2 mL of hydrochloric acid, and again boil for 2 min. Cool, filter, and wash the filter with water until the filtrate measures 100 mL. To 10 mL of the filtrate add 1 mL of 2 N sulfuric acid: no turbidity is produced. HEAVY METALS, Method II 231: 20 ppm SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 231242, but the range between beginning and end of melting does not exceed 3. OPTICAL ROTATION, Specific Rotation 781S: +29.0 to +32.5 Sample solution: 20 mg/mL, in dioxane LOSS ON DRYING 731: Dry at 105 for 2 h: it loses NMT 1.0% of its weight. MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the test for absence of Salmonella species. ODOR ON BOILING: Boil 2 g with 100 mL of water for 2 min: the mixture is odorless. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dehydrocholic Acid RS
PERFORMANCE TESTS DISINTEGRATION 701 Time: 30 min UNIFORMITY OF DOSAGE UNITS 905:
SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: Meet the requirements of the test for absence of Salmonella species ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dehydrocholic Acid RS
Demecarium Bromide
(Comment on this Monograph)id=m22640=Demecarium Bromide=D-Monos.pdf)
Dehydrocholic Acid Tablets

(Comment on this Monograph)id=m22590=Dehydrocholic Acid Tablets=D-Monos.pdf) DEFINITION Dehydrocholic Acid Tablets contain NLT 94.0% and NMT 106.0% of the labeled amount of C24H34O5. IDENTIFICATION PROCEDURE Sample: Mix a quantity of finely powdered Tablets equivalent to 500 mg of dehydrocholic acid with 15 mL of water, and add slowly, with stirring, 2 mL of sodium carbonate TS. Filter, and add to the filtrate 3 N hydrochloric acid (2 mL) dropwise until no more precipitate is formed. Filter the precipitate, wash with small portions of cold water until free from chloride, and dry at 105 for 2 h. Use the residue for the following: Analysis 1: Infrared Absorption 197K Acceptance criteria 1: Meets the requirements Analysis 2: Melting Range or Temperature 741 Acceptance criteria 2: 231242, but the range between beginning and end of melting does not exceed 3 ASSAY PROCEDURE Sample: Weigh and finely powder NLT 20 Tablets. Transfer a portion of the powder nominally equivalent to 500 mg of dehydrocholic acid to a 300-mL conical flask. Analysis: Add 60 mL of neutralized alcohol, and warm on a steam bath for 10 min. Cool, and add phenolphthalein TS and 20 mL of water. Titrate with 0.1 N sodium hydroxide VS, adding 100 mL of water shortly before the endpoint is reached. Each mL of 0.1 N sodium hydroxide is equivalent to 40.25 mg of C24H34O5.
716.59 C32H52Br2N4O4 Benzenaminium, 3,3-[1,10-decanediylbis [(methylimino)carbonyloxy]]bis[N,N,N-trimethyl]-, dibromide; (m-Hydroxyphenyl)trimethylammonium bromide decamethylenebis[methylcarbamate] (2:1) [56-94-0]. DEFINITION Demecarium Bromide contains NLT 95.0% and NMT 100.5% of C32H52Br2N4O4, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solution: 2 mg/mL in 1 N sodium hydroxide Analysis: Reflux 50 mL of Sample solution for 15 min. Cool, and add 3 mL of the refluxed solution to 25 mL of saturated sodium bicarbonate solution. Add, with mixing, 4 mL of N,N-dimethyl-p-phenylenediamine dihydrochloride solution (1.5 in 10,000) and 2 mL of sodium hypochlorite solution (1.5 in 20,000). Acceptance criteria: A violet-blue color is produced. C. PROCEDURE Sample solution: Dissolve 50 mg in 20 mL of water Analysis: To the Sample solution, add 10 mL of a 1-in-50 solution of ammonium reineckate in methanol, and allow to stand for 30 min with occasional swirling. Acceptance criteria: A pink reineckate of demecarium forms, and it melts between 131 and 136, with decomposition. D. IDENTIFICATION TESTSGENERAL, Bromide 191 ASSAY PROCEDURE Sample: 0.8 g of Demecarium Bromide Analysis: Dissolve in glacial acetic acid and mercuric acetate TS (75:15), warming slightly, if necessary, to dissolve. Add 2 drops of crystal violet TS and titrate with 0.1 N perchloric acid VS. Perform a blank determination, and make any
USP 32
necessary correction. Each mL of 0.1 N perchloric acid is equivalent to 35.83 mg of C32H52Br2N4O4. Acceptance criteria: 95.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method I 231: NMT 20 ppm Organic Impurities PROCEDURE: LIMIT OF m-TRIMETHYLAMMONIOPHENOL BROMIDE Control solution: Dissolve 100 mg of mdimethylaminophenol in 10 mL of alcohol in a 1000-mL volumetric flask, dilute with water to volume, and mix. Pipet 1 mL of this solution into a 500-mL volumetric flask, dilute with water to volume, and mix to obtain a 0.2 g/mL solution of m-dimethylaminophenol. Sample solution: 1 mg/mL of Demecarium Bromide in water Analysis: Pipet 25 mL of the Sample solution into a glassstoppered, 50-mL centrifuge tube, and pipet 25 mL of the Control solution into a second, similar tube. To each tube add 3 mL of pH 7.0 Phosphate Buffer (see Reagents, Indicators, and SolutionsBuffer Solutions), 1 mL of N,Ndimethyl-p-phenylenediamine dihydrochloride solution (1.5 in 10,000), 5 mL of isobutyl alcohol, and 1 mL of sodium hypochlorite solution (1.5 in 20,000). Insert the stoppers in the tubes, shake the mixtures for 5 min, and centrifuge. Acceptance criteria: Any blue color produced in the upper layer obtained from the Sample solution is not more intense than that obtained from the Control solution. SPECIFIC TESTS PH 791: 5.07.0, in a solution 1 in 100 WATER DETERMINATION, Method I 921: NMT 2.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Demecarium Bromide RS
Official Monographs / Demeclocycline 17

C. IDENTIFICATION TESTSGENERAL, Bromide 191: requirements Meets the
ASSAY PROCEDURE Sample solution: Ophthalmic Solution Standard solution: 0.5 mg/mL of USP Demecarium Bromide RS Spectrometric conditions Mode: UV Analytical wavelength: 292 nm Analysis Samples: Standard solution and Sample solution Into each of two 50-mL volumetric flasks marked 1 and 2 pipet the Sample solution to nominally contain 2.5 mg of demecarium bromide; into each of two additional 50-mL volumetric flasks, marked 3 and 4, pipet 5.0 mL of a freshly prepared Standard solution. To flasks 1 and 3, add 1 N sodium hydroxide to volume. Transfer 10.0 mL of each of these solutions to separate glass-stoppered tubes, insert the stoppers loosely, heat the tubes in a water bath for 15 min, then cool to room temperature. The concentration of USP Demecarium Bromide RS in the Standard solution is 50 g/mL. To flasks 2 and 4, add pH 10.0 alkaline borate buffer (see Reagents, Indicators, and SolutionsSolutions) to volume. Concomitantly determine the absorbances of the two sodium hydroxide solutions (flasks 1 and 3) using the corresponding borate buffer solutions (flasks 2 and 4) as the respective solvent blanks. Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Demecarium Bromide RS in the Standard solution (mg/mL) CU = nominal concentration of demecarium bromide in the Sample solution (mg/mL) Acceptance criteria: 92.0%108.0% SPECIFIC TESTS STERILITY TESTS 71: Meets the requirements AU AS CS
Demecarium Bromide Ophthalmic Solution

(Comment on this Monograph)id=m22650=Demecarium Bromide Ophthalmic Solution=D-Monos.pdf) DEFINITION Demecarium Bromide Ophthalmic Solution is a sterile, aqueous solution of Demecarium Bromide. It contains NLT 92.0% and NMT 108.0% of the labeled amount of C32H52Br2N4O4. It contains a suitable antimicrobial agent. IDENTIFICATION A. PROCEDURE Sample solution: Ophthalmic Solution and 1 N sodium hydroxide (10:12.5) Analysis: Reflux for 15 min. Cool, and add 3 mL of the refluxed solution to 25 mL of saturated sodium bicarbonate solution. Add 4 mL of N,N-dimethyl-p-phenylenediamine dihydrochloride solution (0.15 mg/mL) and 2 mL of sodium hypochlorite solution (0.075 mg/mL). Acceptance criteria: A violet-blue color is produced. B. PROCEDURE Sample solution: 10 mL of solution Analysis: To the Sample solution add 5 mL solution (20 mg/mL) of ammonium reineckate in methanol, and allow to stand for 30 min with occasional swirling. Acceptance criteria: A pink reineckate of demecarium forms, and it melts between 131 and 136, with decomposition.
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Demecarium Bromide RS
Demeclocycline
(Comment on this Monograph)id=m22680=Demeclocycline=DMonos.pdf)
464.85 C21H21ClN2O8 2-Naphthacenecarboxamide, 7-chloro-4(dimethylamino)-1,4,4a,5,5a,6,11,12aoctahydro-3,6,10,12,12a-pentahydroxy-1,11-dioxo-, [4S(4,4a,5a,6,12a)]-; 7-Chloro-4-(dimethylamino)-1,4,4a,5,5a,6,11,12aoctahydro-3,6,10,12,12a-pentahydroxy-1,11-dioxo-2naphthacenecarboxamide [127-33-3]. Sesquihydrate 491.88 [13215-10-6].
18
Demeclocycline / Official Monographs
USP 32
Sample solution: 0.9 mg/mL of Demeclocycline in 0.01 N hydrochloric acid System suitability solution: USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid. Allow to stand for 3 h. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L21 Column temperature: 60 0.5 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for epidemethylchlortetracycline and demeclocycline are 0.7 and 1.0, respectively, System suitability solution] Suitability requirements Resolution: NLT 3.0 between the epidemethylchlortetracycline and demeclocycline peaks, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C21H21ClN2O8 HCl equivalent in each mg of Demeclocycline taken: Result = (rU/rS) (CS/W) V P = peak response area of demeclocycline from the Sample solution = peak response area of demeclocycline from the rS Standard solution = concentration of USP Demeclocycline CS Hydrochloride RS in the Standard solution (mg/mL) W = quantity of the Demeclocycline taken (mg) V = volume of Sample solution (mL) P = demeclocycline hydrochloride equivalent of the USP Demeclocycline Hydrochloride RS taken (g/mg) Acceptance criteria: Potency equivalent to NLT 970 g/mg of C21H21ClN2O8 HCl SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 4.05.5, in 10 mg/mL solution WATER DETERMINATION, Method I 921: 4.3%6.7% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Demeclocycline Hydrochloride RS rU
DEFINITION Demeclocycline has a potency equivalent to NLT 970 g of demeclocycline hydrochloride (C21H21ClN2O8 HCl) per mg, calculated on the anhydrous basis. IDENTIFICATION A. PROCEDURE Sample solution: Transfer 40 mg into a 250-mL volumetric flask, add 2 mL of 0.1 N hydrochloric acid to dissolve, and dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, add about 75 mL of water and 5.0 mL of 5 N sodium hydroxide, dilute with water to volume, and mix. Acceptance criteria: The UV absorption spectrum of this solution, measured at 6 min, accurately timed, after the addition of the sodium hydroxide, exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Demeclocycline Hydrochloride RS, concomitantly measured, and the absorptivity, calculated on the anhydrous basis, at the wavelength of maximum absorbance at about 380 nm is between 103.5% and 111.3% of that of the USP Demeclocycline Hydrochloride RS, the potency of the Reference Standard being taken into account. B. PROCEDURE Solutions: Transfer 40 mg to a 200-mL volumetric flask, add 100 mL of 0.1 N hydrochloric acid, shake to dissolve, and dilute with 0.1 N hydrochloric acid to volume. Transfer 5.0 mL of this solution into each of two 50-mL volumetric flasks (Solutions 1 and 2). Prepare similar solutions of USP Demeclocycline Hydrochloride RS (Solutions 3 and 4). To Solutions 1 and 3, add 10 mL of 6 N hydrochloric acid, and to Solutions 2 and 4, add 10 mL of 3 N hydrochloric acid. Heat the four flasks in a water bath for 20 min, cool, and dilute the contents with water to volume. Analysis: Determine the absorbances of Solutions 1 and 3 at the wavelength of maximum absorbance at about 430 nm, with a suitable spectrophotometer, using Solutions 2 and 4, respectively, as the blanks. Determine the absorbances of Solutions 2 and 4 at the wavelength of maximum absorbance at about 368 nm, using Solutions 1 and 3, respectively, as the blanks. Calculate the ratio: (WS P/1000) AU/WU AS = weight of USP Demeclocycline Hydrochloride RS taken, calculated on the dried basis (mg) P = potency of the USP Demeclocycline Hydrochloride RS (g/mg) = absorbence of A368 + A430 for the specimen (U) AU = weight of specimen taken, calculated on the WU anhydrous basis (mg) AS = absorbence of A368 + A430 for the Standard (S) Acceptance criteria: 0.971.17 WS ASSAY PROCEDURE Mobile phase: Transfer 80 g of tertiary butyl alcohol to a 1000-mL volumetric flask with the aid of 200 mL of water. Add 100 mL of 0.2 M pH 9.0 phosphate buffer (prepared by mixing appropriate volumes of 0.2 M dibasic potassium phosphate and 0.2 M monobasic potassium phosphate), 150 mL of 0.02 M tetrabutylammonium hydrogen sulfate (adjusted with sodium hydroxide TS to a pH of 9.0), and 100 mL of 0.01 M edetate disodium (adjusted with sodium hydroxide TS to a pH of 9.0). Dilute with water to volume and degas. Decreasing the amount of tertiary butyl alcohol increases the resolution. Standard solution: 1 mg/mL of USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid
Demeclocycline Oral Suspension

(Comment on this Monograph)id=m22690=Demeclocycline Oral Suspension=D-Monos.pdf) DEFINITION Demeclocycline Oral Suspension contains the equivalent of NLT 90.0% and NMT 125.0% of the labeled amount of demeclocycline hydrochloride (C21H21ClN2O8 HCl). It may contain one or more suitable buffers, preservatives, stabilizers, and suspending agents.
USP 32
IDENTIFICATION IDENTIFICATIONTETRACYCLINES, Method II 193 Sample solution: 1 mg/mL of demeclocycline hydrochloride, from Oral Suspension in methanol. Shake and allow the mixture to settle. Use the clear supernatant. ASSAY PROCEDURE Mobile phase: Transfer 80 g of tertiary butyl alcohol to a 1000-mL volumetric flask with the aid of 200 mL of water, add 100 mL of 0.2 M pH 9.0 phosphate buffer (prepared by mixing appropriate volumes of 0.2 M dibasic potassium phosphate and 0.2 M monobasic potassium phosphate), 150 mL of 0.02 M tetrabutylammonium hydrogen sulfate (adjusted with sodium hydroxide TS to a pH of 9.0), and 100 mL of 0.01 M edetate disodium (adjusted with sodium hydroxide TS to a pH of 9.0). Dilute to volume and degas. Decreasing the amount of tertiary butyl alcohol increases the resolution. Standard solution: 1 mg/mL of USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid Sample solution: Oral Suspension, freshly mixed and free from air bubbles, nominally equivalent to 50 mg of C21H21ClN2O8 HCl, to a 50-mL volumetric flask, dilute with 0.01 N hydrochloric acid to volume. Sonicate for 5 min, and centrifuge for 5 min. Pass a portion of the supernatant through a suitable filter having a 1.5-m or finer porosity, and use the clear filtrate. System suitability solution: Prepare a solution of USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid, and allow to stand for 3 h. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L21 Column temperature: 60 0.5 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for epidemethylchlortetracycline and demeclocycline are about 0.7 and 1.0, respectively, System suitability solution] Suitability requirements Resolution: NLT 3.0 between the epidemethylchlortetracycline peak and the demeclocycline peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H21ClN2O8 HCl equivalent in each mL of Oral Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Demeclocycline Hydrochloride RS in the Standard solution (mg/mL) CU = nominal concentration of demeclocycline hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%125.0% rU rS CS PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905 For Oral Suspension packaged in single-unit containers:

Meets the requirements DELIVERABLE VOLUME 698 (for Oral Suspension packaged in multiple-unit containers): Meets the requirements SPECIFIC TESTS CRYSTALLINITY 695: PH 791: 4.05.8 Meets the requirements
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light. USP REFERENCE STANDARDS 11 USP Demeclocycline Hydrochloride RS
Demeclocycline Hydrochloride
(Comment on this Monograph)id=m22720=Demeclocycline Hydrochloride=D-Monos.pdf) 501.31 C21H21ClN2O8 HCl 2-Naphthacenecarboxamide, 7-chloro-4(dimethylamino)-1,4,4a,5,5a,6,11,12aoctahydro-3,6,10,12,12a-pentahydroxy-1,11-dioxo-, monohydrochloride, 4S-(4,4a,5a,6,12a)-; 7-Chloro-4-(dimethylamino)-1,4,4a,5,5a,6,11,12aoctahydro-3,6,10,12,12a-pentahydroxy-1,11-dioxo-2naphthacenecarboxamide monohydrochloride [64-73-3]. DEFINITION Demeclocycline Hydrochloride has a potency of NLT 900 g/ mg of C21H21ClN2O8 HCl, calculated on the dried basis. IDENTIFICATION A. PROCEDURE Sample solution: Transfer 40 mg to a 250-mL volumetric flask, add 2 mL of 0.1 N hydrochloric acid to dissolve, and dilute with water to volume. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, add about 75 mL of water and 5.0 mL of 5 N sodium hydroxide, dilute with water to volume, and mix. Acceptance criteria: The UV absorption spectrum of this solution, measured at 6 min, accurately timed, after the addition of the sodium hydroxide, exhibits maxima and minima at the same wavelengths as that of a similar solution of USP Demeclocycline Hydrochloride RS, concomitantly measured. The absorptivity, calculated on the dried basis, at the wavelength of maximum absorbance at about 380 nm, is between 95.8% and 104.2% that of the USP Demeclocycline Hydrochloride RS, the potency of the Reference Standard being taken into account. B. PROCEDURE Solutions: Transfer 40 mg to a 200-mL volumetric flask, add 100 mL of 0.1 N hydrochloric acid, shake to dissolve, and dilute with 0.1 N hydrochloric acid to volume. Transfer 5.0 mL of this solution into each of two 50-mL volumetric flasks (Solutions 1 and 2). Prepare similar solutions of USP Demeclocycline Hydrochloride RS (Solutions 3 and 4). To Solutions 1 and 3, add 10 mL of 6 N hydrochloric acid, and to Solutions 2 and 4, add 10 mL of 3 N hydrochloric acid. Heat the four flasks in a water bath for 20 min, cool, and dilute the contents with water to volume. Analysis: Determine the absorbances of Solutions 1 and 3 at the wavelength of maximum absorbance at about 430 nm, with a suitable spectrophotometer, using Solutions 2 and 4, respectively, as the blanks. Determine the absorbances of Solutions 2 and 4 at the wavelength of maximum absorbance at about 368 nm, using Solutions 1 and 3, respectively, as the blanks.
20

Calculate the ratio: Result = (WS P/1000) (AU/WU) AS P
USP 32
= demeclocycline hydrochloride equivalent of the USP Demeclocycline Hydrochloride RS taken (g/mg) V = volume of Sample solution (mL) Acceptance criteria: Potency of NLT 900 g/mg SPECIFIC TESTS CRYSTALLINITY 695: Meets the requirements PH 791: 2.03.0 in 10 mg/mL solution LOSS ON DRYING 731: Dry 100 mg in a capillary-stoppered bottle in vacuum at a pressure not exceeding 5 mm of mercury at 60 for 3 h: it loses NMT 2.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Demeclocycline Hydrochloride RS
= weight of USP Demeclocycline Hydrochloride RS taken, calculated on the dried basis (mg) P = potency of the USP Demeclocycline Hydrochloride RS (g/mg) = absorbance of A368 + A430 for the specimen (U) AU = weight of specimen taken, calculated on the WU anhydrous basis (mg) = absorbance of A368 + A430 for the Standard (S) AS Acceptance criteria: 0.91.1 WS ASSAY PROCEDURE Mobile phase: Transfer 80 g of tertiary butyl alcohol to a 1000-mL volumetric flask with the aid of 200 mL of water, add 100 mL of 0.2 M pH 9.0 phosphate buffer (prepared by mixing appropriate volumes of 0.2 M dibasic potassium phosphate and 0.2 M monobasic potassium phosphate), 150 mL of 0.02 M tetrabutylammonium hydrogen sulfate (adjusted with sodium hydroxide TS to a pH of 9.0), and 100 mL of 0.01 M edetate disodium (adjusted with sodium hydroxide TS to a pH of 9.0). Dilute with water to volume and degas. Decreasing the amount of tertiary butyl alcohol increases the resolution. Standard solution: 1 mg/mL of USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid Sample solution: 1 mg/mL of Demeclocycline Hydrochloride in 0.01 N hydrochloric acid System suitability solution: USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid Allow to stand for 3 h. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L21 Column temperature: 60 0.5 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for epidemethylchlortetracycline and demeclocycline are 0.7 and 1.0, respectively, System suitability solution.] Suitability requirements Resolution: NLT 3.0 between the epidemethylchlortetracycline peak and the demeclocycline peak, System suitability solution Analysis Samples: Standard solution and Sample solution Calculate the quantity, in g, of C21H21ClN2O8 HCl in each mg of Demeclocycline Hydrochloride taken: Result = (rU/rS) (CS/W) P W rU rS CS W = peak area for demeclocycline from the Sample solution = peak area for demeclocycline from the Standard solution = concentration of USP Demeclocycline Hydrochloride RS in the Standard solution (mg/mL) = quantity of the Demeclocycline taken (mg)
Demeclocycline Hydrochloride Capsules

(Comment on this Monograph)id=m22730=Demeclocycline Hydrochloride Capsules=D-Monos.pdf) DEFINITION Demeclocycline Hydrochloride Capsules contain NLT 90.0% and NMT 125.0% of the labeled amount of demeclocycline hydrochloride (C21H21ClN2O8 HCl). IDENTIFICATION IDENTIFICATIONTETRACYCLINES, Method II 193 Sample solution: 1 mg/mL of demeclocycline hydrochloride, from Capsule contents in methanol. Filter and use the filtrate as Sample solution. ASSAY PROCEDURE Mobile phase: Transfer 80 g of tertiary butyl alcohol to a 1000-mL volumetric flask with the aid of 200 mL of water, add 100 mL of 0.2 M pH 9.0 phosphate buffer (prepared by mixing appropriate volumes of 0.2 M dibasic potassium phosphate and 0.2 M monobasic potassium phosphate), 150 mL of 0.02 M tetrabutylammonium hydrogen sulfate (adjusted with sodium hydroxide TS to a pH of 9.0), and 100 mL of 0.01 M edetate disodium (adjusted with sodium hydroxide TS to a pH of 9.0). Dilute with water to volume and degas. Decreasing the amount of tertiary butyl alcohol increases the resolution. Standard solution: 1 mg/mL of USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid Sample solution: Remove, as completely as possible, the contents of NLT 10 Capsules, and weigh. Mix, and transfer a portion of the powder, nominally equivalent to 50 mg of demeclocycline hydrochloride, to a 50-mL volumetric flask, and dilute with 0.01 N hydrochloric acid to volume. Sonicate for 5 min, and centrifuge for 5 min. Pass a portion of the supernatant through a suitable filter having a 1.5-m or finer porosity, and use the clear filtrate. System suitability solution: A solution of USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid Allow to stand for 3 h. Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L21 Column temperature: 60 0.5 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: Standard solution and System suitability solution [NOTEThe relative retention times for epidemethylchlortetracycline and demeclocycline are 0.7 and 1.0, respectively, System suitability solution] Suitability requirements Resolution: NLT 3.0 between the epidemethylchlortetracycline peak and the demeclocycline peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H21ClN2O8 HCl in the portion of Capsule contents taken: Result = (rU/rS) (CS/CU) 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Demeclocycline Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of demeclocycline CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%125.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 45 min Standard solution: USP Demeclocycline Hydrochloride RS at a known concentration in Medium Sample solution: Sample per Dissolution 711. Dilute with Medium as needed. Spectrometric conditions Mode: UV Analytical wavelength: 274 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C21H21ClN2O8 UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS LOSS ON DRYING 731: Dry 100 mg of Capsule contents in a capillary-stoppered bottle in vacuum at 60 for 3 h: the material loses NMT 2.0% of its weight, except that if the Capsules contain starch, the material loses NMT 8.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Demeclocycline Hydrochloride RS
Demeclocycline Hydrochloride Tablets

(Comment on this Monograph)id=m22740=Demeclocycline Hydrochloride Tablets=D-Monos.pdf) DEFINITION Demeclocycline Hydrochloride Tablets contain NLT 90.0% and NMT 125.0% of the labeled amount of demeclocycline hydrochloride (C21H21ClN2O8 HCl). IDENTIFICATION IDENTIFICATIONTETRACYCLINES, Method II 193 Sample solution: 1 mg/mL of demeclocycline hydrochloride, from finely powdered Tablets in methanol Use the filtrate. ASSAY PROCEDURE Mobile phase: Transfer 80 g of tertiary butyl alcohol to a 1000-mL volumetric flask with the aid of 200 mL of water. Add 100 mL of 0.2 M pH 9.0 phosphate buffer (prepared by mixing appropriate volumes of 0.2 M dibasic potassium phosphate and 0.2 M monobasic potassium phosphate), 150 mL of 0.02 M tetrabutylammonium hydrogen sulfate (adjusted with sodium hydroxide TS to a pH of 9.0), and 100 mL of 0.01 M edetate disodium (adjusted with sodium hydroxide TS to a pH of 9.0). Dilute with water to volume, and degas. Decreasing the amount of tertiary butyl alcohol increases the resolution. Standard solution: 1 mg/mL of USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid Sample solution: Weigh and finely powder NLT 10 Tablets. Transfer a portion, nominally equivalent to 50 mg demeclocycline hydrochloride, to a 50-mL volumetric flask, dilute with 0.01 N hydrochloric acid to volume. Sonicate for 5 min, and centrifuge for 5 min. Pass a portion of the supernatant through a suitable filter of 1.5-m or finer porosity, and use the clear filtrate. System suitability solution: USP Demeclocycline Hydrochloride RS in 0.01 N hydrochloric acid. Allow to stand for 3 h. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L21 Column temperature: 60 0.5 Flow rate: 1 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for epidemethylchlortetracycline and demeclocycline are about 0.7 and 1.0, respectively, System suitability solution.] Suitability requirements Resolution: NLT 3.0 between the epidemethylchlortetracycline peak and the demeclocycline peak, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution
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derived, test negative for human and animal viruses and retroviruses and are also tested for normal cell morphology, human karyology, and isoenzymes. Maternal blood sera are tested for evidence of infection with human immunodeficiency virus types 1 and 2, hepatitis B and C viruses, syphilis, and human T-lymphotropic virus type 1 and are found negative for the purpose of donor selection. Cryopreserved Human Fibroblast-Derived Dermal Substitute is manufactured with sterile components under aseptic conditions within the final package. Reagents used in the manufacture of Cryopreserved Human Fibroblast-Derived Dermal Substitute are tested and found free of viruses, retroviruses, endotoxins, and mycoplasma before use. All materials derived from bovine sources originate from countries free of bovine spongiform encephalopathy. During subsequent screening of the fibroblast cell strain at various stages in the manufacturing process, testing for these same viruses, as well as Epstein-Barr virus and human T-lymphotropic virus type 2, is carried out and found to be negative. The final product tests negative for the presence of mycoplasma. SPECIFIC TESTS HISTOLOGICAL EVALUATION Buffered Formalin: Prepare a solution containing 10% (w/v) formaldehyde solution and 1.0% to 1.5% methanol in a suitable buffer, adjusted to a pH of 6.8 to 7.2.1 Preparation of Tissue for Staining: Cut Cryopreserved Human Fibroblast-Derived Dermal Substitute into 3-mm 3mm sections. Place three sections into suitable histological tissue cassettes,2 and insert the cassettes into suitable histological cassette basket(s).3 At a temperature of 40, sequentially immerse the histological cassette basket(s) in separate solutions of Buffered formalin (2 h), two changes of 80% alcohol (30 min/step), alcohol (30 min), three changes of dehydrated alcohol (30 min/step), suitable histological xylene substitute (30 min),4 and two changes of suitable xylene substitute (30 min/step). Immerse the histological cassette basket(s) into molten paraffin5 that is at a temperature of 60 for 30 min. Remove the cassette basket(s), and immerse in a fresh container of molten paraffin (60) for 60 min. Remove the histological tissue cassette from the container and basket, and disassemble. Fill preheated embedding molds with molten paraffin heated to 56 to 60, and place on top of a preheated warming platform that is designed for histology work. Transfer Cryopreserved Human Fibroblast-Derived Dermal Substitute specimens from the cassettes using forceps, and place specimens into individual molds. Orient the specimens in molds so as to cut cross-sections. Cool the paraffin by sliding the mold down the platform to its cool side until the paraffin has solidified. Maintain specimen orientation with forceps during cooling, removing the forceps when the paraffin becomes translucent. Slide the paraffin block onto a histological cold plate to rapidly cool the block. Trim the paraffin block with a new single-edged razor blade to form a rectangle or slight trapezoid to within 5 mm of the tissue
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Analysis Samples: Standard solution and Sample solution Calculate the percentage of C21H21ClN2O8 HCl in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak area of the Sample solution = peak area of the Standard solution = concentration of USP Demeclocycline Hydrochloride RS in the Standard solution (mg/mL) = nominal concentration of demeclocycline CU hydrochloride in the Sample solution (mg/mL) Acceptance criteria: 90.0%125.0% rU rS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 900 mL Apparatus 2: 75 rpm Time: 45 min Detector: UV 274 nm Sample solution: Sample per Dissolution 711. Dilute with Medium as needed. Standard solution: USP Demeclocycline Hydrochloride RS at a known concentration in Medium Spectrometric conditions Mode: UV Analytical wavelength: 274 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C21H21ClN2O8 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS LOSS ON DRYING 731: Dry 100 mg of finely ground Tablet powder in a capillary-stoppered bottle in vacuum at 60 for 3 h: it loses NMT 2% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Demeclocycline Hydrochloride RS
Cryopreserved Human FibroblastDerived Dermal Substitute

(Comment on this Monograph)id=m33155=Cryopreserved Human Fibroblast-Derived Dermal Substitute=D-Monos.pdf) DEFINITION Cryopreserved Human Fibroblast-Derived Dermal Substitute is a living monolayer skin substitute derived from neonatal foreskins. It is composed of fibroblasts, an extracellular matrix, and a bioabsorbable scaffold. Human fibroblasts are seeded onto a bioabsorbable, nonantigenic and nonpyrogenic mesh scaffold composed of polyglactin, a copolymer of glycolide and lactide. The fibroblasts proliferate to fill the interstices of this scaffold to create a three-dimensional human dermal substitute that secretes human dermal collagen, matrix proteins, growth factors, and cytokines. Cryopreserved Human Fibroblast-Derived Dermal Substitute does not contain macrophages, lymphocytes, blood vessels, hair follicles, muscle fibers, or keratin. The fibroblast-cell banks, from which Cryopreserved Human Fibroblast-Derived Dermal Substitute is
suitable Buffered formalin can be obtained from VWR International, 1310 Goshen Pkwy., West Chester, PA 19380. 2A suitable histological tissue cassette can be obtained from Sakura Finetek U.S.A., Inc., 1750 West 214th St., Torrance, CA 90501. 3suitable histological tissue cassette basket can be obtained from Sakura Finetek U.S.A., Inc., 1750 West 214th St., Torrance, CA 90501. 4A suitable histological xylene substitute is Citrosolve Clearing Agent, available from Fisher Scientific, 200 Park Ln., Pittsburgh, PA 15275. 5A suitable paraffin for use is Tissue Prep* 2 Embedding Media, available from Fisher Scientific, 200 Park Ln., Pittsburgh, PA 15275.
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mass, if necessary. Cool the block at 4 for 15 to 30 min. Clamp the tissue block into the block holder of the microtome. Fill a histological tissue flotation water bath with fresh water, add an appropriate amount of a suitable histological adhesive,6 and heat to 5 less than the melting point of the paraffin. Properly mount and adjust the tissue and paraffin block into a microtome. Set the microtome to make cuts 5 thick with a blade angle of 5 2. Insert a sharp stainless steel microtome knife that has been properly honed or a new disposable microtome knife into the knife holder. Cut a ribbon that contains 6 to 10 sections of Cryopreserved Human Fibroblast-Derived Dermal Substitute. Pick up the ribbon with forceps, and stretch it across the tissue flotation water bath. Separate 2 to 3 adjacent sections from the ribbon on the water bath. The selected sections should not be compressed, wrinkled, or scratched. Pick up the selected sections by dipping a microscope slide into the water bath under the floating sections, and gently lift the slide out of the water. For each staining procedure, prepare three slides from each of the three starting Cryopreserved Human Fibroblast-Derived Dermal Substitute 3-mm 3-mm sections. Allow the mounted sections to air-dry completely, or dry the slide in a 60 oven for 1 h. HematoxylinEosin Staining Hematoxylinalcohol solution: Dissolve 2.5 g of hematoxylin in 25.0 mL of dehydrated alcohol with heating. Potassium alum solution: Dissolve 50.0 g of potassium alum in 500 mL of water with heating. Hematoxylin staining solution: Mix Hematoxylinalcohol solution and Potassium alum solution. Bring to a boil as rapidly as possible with constant stirring. Do not heat for more than 1 min. Slowly add 0.185 g of sodium iodate. Reheat to a simmer until the solution becomes a deep purple. Remove from heat, and quickly cool. Filter daily before use. 10% acid alcohol: Add 5.0 mL of hydrochloric acid to 495 mL of 70% alcohol. Eosin solution: Dissolve 1 g of eosin Y in 100 mL of alcohol. Filter daily before use. Analysis: The microscope slide with affixed tissue, as prepared in Preparation of Tissue for Staining, is sequentially immersed in three changes of a suitable histological, aliphatic xylene substitute (2 min/step), three changes of dehydrated alcohol (1 min/step), alcohol (20 s), running tap water rinse (1 min), Hematoxylin staining solution (4 to 5 min), running tap water rinse (1 min), 10% acid alcohol (15 s), running tap water rinse (1 min), a suitable bluing reagent7 (20 to 30 s), running tap water rinse (1 min), alcohol (20 s), Eosin solution (10 to 20 s, until a reddishbrown color is obtained in the tissue), three changes of dehydrated alcohol (10 s/step), and three changes of a suitable histological xylene substitute (10 s/step). Adjust the above immersion times as needed to suitably stain the tissue. Remove the slide from the last histological xylene substitute wash, and blot dry the back of the slide. Do not allow the tissue to dry. Affix a coverslip over the tissue using a coverslip mountant. Using USP Cryopreserved Human Fibroblast-Derived Dermal Substitute Reference Photomicrograph 1 for comparison, the test article shows a polyglactin scaffold mesh and a secreted collagen-based matrix; the tissue contains normal human fibroblast distributed throughout the secreted matrix and resembles normal human papillary dermis. Fibroblasts appear elongated and spindle-shaped. The tissue, which is 100 to 300 m thick, contains 106 cells/cm2.
suitable histological adhesive for use is Histoslide Adhesive, which can be obtained from Poly Scientific Research Corp., 70 Cleveland Ave., Bay Shore, NY 11706-1282. 7A suitable bluing reagent can be obtained from Sigma-Aldrich Corp., P.O. Box 14508, St. Louis, MO 63178.
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Collagen Staining Bouinss solution: Mix 75 mL of 1.22% picric acid solution, 25 L of dimethoxymethane, and 5.0 L of acetic acid. Weigerts iron hematoxylin solution A: 1 g of hematoxylin in 100 mL of alcohol Weigerts iron hematoxylin solution B: Mix 4 mL of 29% ferric chloride, 95 mL of water, and 1 mL of hydrochloric acid. Weigerts iron hematoxylin working solution: Mix equal volumes of Weigerts iron hematoxylin solution A and Weigerts iron hematoxylin solution B. Pass the solution through a filter having a 0.45-m porosity. Prepare fresh as needed. Gomoris trichrome solution: Mix 1.0 mL of acetic acid and 100 mL of water. Dissolve 0.6 g of chromotrope 2R, 0.3 g of Fast Green FCF, and 0.6 g of phosphotungstic acid. 1% acetic acid: 1 mL of acetic acid and 100 mL of water Analysis: The microscope slide with affixed tissue, as prepared in Preparation of Tissue for Staining, is sequentially immersed in three changes of a suitable histological, aliphatic xylene substitute (2 min/step), three changes of dehydrated alcohol (1 min/step), alcohol (20 s), and running tap water rinse (1 min). Immerse the slide in Bouinss solution, and place in a 42 water bath for 1 h. Rinse the slide in water for 10 s. Sequentially immerse the slide in Weigerts iron hematoxylin working solution (10 min) and running tap water rinse (10 min). Rinse the slide in water for 10 s, and immerse in Gomoris trichrome solution (3 to 5 min). Rinse the slide in 1% acetic acid for 20 s. Sequentially immerse the slide in three changes of alcohol (10 s/step) and three changes of a suitable histological, aliphatic xylene substitute (10 s/step). Affix a coverslip over the tissue using a suitable coverslip mountant. Nuclei will stain black; cytoplasm, keratin, and muscle fibers will stain red; and collagen and mucin will stain blue. Using USP Cryopreserved Human Fibroblast-Derived Dermal Substitute Reference Photomicrograph 2 for comparison, the tissue contains normal human fibroblast distributed throughout the secreted matrix and resembles normal human papillary; dermis, muscle fibers, and keratin are absent. Distribution of Fibronectin Tris-saline buffer: Prepare a solution containing 0.1 M tris (hydroxymethyl) aminomethane hydrochloride and 0.15 M sodium chloride, adjusted to a pH of 7.8. 3% hydrogen peroxide: 30 mL of hydrogen peroxide in water or methanol Diaminobenzidine solution: Use a suitable solution.8 Hematoxylin staining solution: Prepare as directed for HematoxylinEosin Staining. Analysis: The microscope slide with affixed tissue as prepared in Preparation of tissue for staining is dried either overnight at 37 or for 1 h at 60. The microscope slide with affixed tissue as prepared in Preparation of Tissue for Staining is sequentially immersed in three changes of a suitable histological, aliphatic xylene substitute (2 min/step), three changes of dehydrated alcohol (1 min/step), alcohol (20 s), and running tap water rinse (1 min). Sequentially immerse the slide in Tris-saline buffer (10 min), 3% hydrogen peroxide (30 min), three changes of Trissaline buffer (1 min/step), a suitable normal rabbit serum9 (30 min), water (5 min), and three changes of Tris-saline buffer (1 min/step). Incubate the slide with a suitable solution of rabbit anti-human fibronectin antibody,10
8A suitable Diaminobenzidine solution can be obtained from Sigma-Aldrich Corp., P.O. Box 14508, St. Louis, MO 63178; catalog number D-6815. 9A suitable normal rabbit serum can be obtained from Dako Corp., 6392 Via Real, Carpinteria, CA 93013. 10Suitable rabbit anti-human fibronectin antibodies can be obtained from Dako Corp., 6392 Via Real, Carpinteria, CA 93013.
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Component
L-Phenylalanine L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine D-Calcium
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mg/L 66.0 42.0 95.2 16.0 72.0 93.6 pantothenate 4.0 4.0 4.0 7.0 4.0 4.0 0.40 4.0
diluted using a suitable antibody diluent11 to an antibody titer of 21.0 2.1 mg/L for 1 h. Sequentially immerse the slide in water (5 min) and three changes of Tris-saline buffer (1 min/step). Place enough drops of a biotinylated goat anti-rabbit antibody solution12 to cover the tissue section, and incubate for 30 min. Sequentially immerse the slide in water (5 min) and three changes of Tris-saline buffer (1 min/step). Place enough drops of a streptavidin conjugated horseradish peroxidase solution13 to cover the tissue section, and incubate for 30 min. Sequentially immerse the slide in water (5 min) and three changes of Tris-saline buffer (1 min/step). Incubate the slide with Diaminobenzidine solution for 1 to 5 min, until a suitable difference in staining is seen by comparison with a control in which the fibronectin (primary) antibody is omitted. Sequentially immerse the slide in water (1 min), Hematoxylin staining solution (4 to 5 min), and water (1 min). Do not allow the tissue to dry. Affix a coverslip over the tissue using a lowviscosity, aqueous, synthetic-resin coverslip mountant. Using USP Cryopreserved Human Fibroblast-Derived Dermal Substitute Reference Photomicrograph 3 for comparison, fibronectin is found colocalizing with the collagen fibers. The intensity of staining may vary from region to region of the slide. METABOLIC ACTIVITY ASSESSMENT DPBS solution A: Dissolve 1.32 g of calcium chloride and 1.21 g of magnesium sulfate heptahydrate in 2 L of water. DPBS solution B: Dissolve 80 g of sodium chloride; 2 g of potassium chloride; 11.5 g of dibasic sodium phosphate; 2 g of monobasic potassium phosphate; 10 g of glucose; 0.36 g of sodium phosphate; 0.5 g of streptomycin sulfate; and 1,000,000 USP Units of penicillin G sodium in 8 L water. DPBS working solution: Mix DPBS solution B with DPBS solution A (8:2). Pass the solution through a filter having a 0.22-m porosity. Dulbeccos modified Eagles tissue culture medium: Prepare a solution that contains the following components:
Component Calcium chloride Ferric nitrate nonahydrate Potassium chloride Magnesium sulfate heptahydrate Sodium chloride Sodium bicarbonate Sodium phosphate, monobasic (monohydrate) Dextrose Phenol red Sodium pyruvate
L-Arginine L-Cystine
Choline chloride Folic acid Inositol Nicotinamide Pyridoxine hydrochloride Riboflavin Thiamine hydrochloride
mg/L 264.9 0.10 400.0 200.0 6,400.0 3,700.0 125.0 4,500.0 15.0 110.0 84.0 48.0 30.0 42.0 104.8 104.8 146.2 30.0
hydrochloride
Aminoacetic acid
L-Histidine L-Leucine L-Lysine
hydrochloride monohydrate
L-Isoleucine
hydrochloride
L-Methionine
11Suitable
antibody diluent can be obtained from Dako Corp., 6392 Via Real, Carpinteria, CA 93013. 12Suitable biotinylated goat anti-rabbit antibody solution can be obtained from BioGenex, 4600 Norris Canyon Rd., San Ramon, CA 94583. 13A suitable streptavidin conjugated horseradish peroxidase solution can be obtained from BioGenex, 4600 Norris Canyon Rd., San Ramon, CA 94583.
L-Glutamine solution: Prepare a 100-mL solution containing 2.92 g of L-glutamine. Sodium pyruvate solution: Prepare 100 mL of a solution containing 1.10 g of sodium pyruvate. Antibiotic-antimycotic solution: Prepare 100 mL of a solution containing 0.85 g of sodium chloride, 10,000 USP Units of penicillin G sodium, 10,000 g of streptomycin (base), and 25 g of amphotericin B. Sample stock medium: Mix 1000 mL of Dulbeccos modified Eagles tissue culture medium, 10 mL of L-Glutamine solution, 10 mL of Sodium pyruvate solution, 10 mL of Antibioticantimycotic solution, and 20 mL of fetal bovine serum.14 MTT-sample solution: Dissolve 0.50 g of 3-(4,5dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide in 1 L of Sample stock medium, using constant stirring. Sterilize the solution by passing it through a suitable filter having a 0.2m porosity. MTT formazan stock solution: Prepare a solution containing 100 g of 1-(4,5-dimethylthiazol-2-yl)-3,5diphenylformazan/mL of isopropyl alcohol. MTT formazan calibration solutions: Prepare solutions containing the following five MTT formazan concentrations: 15 g/mL, 30 g/mL, 45 g/mL, 60 g/mL, and 75 g/mL of MTT formazan, using MTT formazan stock solution and diluting with isopropyl alcohol. Analysis: Thaw Cryopreserved Human Fibroblast-Derived Dermal Substitute by placing the tissue, still in its ethyl vinyl acetate bag, in a water bath heated to between 34 and 37 for 2 to 3 min. The minimum amount of water in the water bath is 2 L/Cryopreserved Human Fibroblast-Derived Dermal Substitute unit. Cut three 11-mm 11-mm sections of Cryopreserved Human Fibroblast-Derived Dermal Substitute, and immerse the sections into separate, 3-mL portions of MTT-sample solution. Incubate for 2 h at 37 2 in a 3% to 7% CO2air environment with shaking on an orbital shaker at 150 to 200 rpm. After incubation remove from the 37, 3% to 7% CO2air environment. Remove the MTT-sample solution, and rinse twice with DPBS working solution. Immerse the Cryopreserved Human Fibroblast-Derived Dermal Substitute in 2 mL of isopropyl alcohol, and incubate at ambient temperature for 1 h with shaking on an orbital shaker at approximately 125 rpm. Transfer 200-L aliquots of the five MTT formazan calibration solutions, in triplicate, and 200-L aliquots of the three isopropyl alcohol extracts of Cryopreserved Human Fibroblast-Derived Dermal Substitute to a suitable 96-well flat-bottom plate. Read the absorbance of each aliquot at 540 nm, using 200 L of isopropyl alcohol as the blank. Plot the responses of the MTT
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suitable fetal bovine serum can be obtained from HyClone, 925 West 1800 South, Logan, UT 84321; catalog number SH30070.03.
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formazan calibration solutions versus concentration, in g/mL of MTT formazan, and calculate the regression line using the least-squares method. The test is considered valid if the regression line has a square of the correlation coefficient NLT 0.95. Acceptance criteria: The absorbance value of individual, thawed, Cryopreserved Human Fibroblast-Derived Dermal Substitute sections at 540 nm is between 0.30 and 0.86. DNA CONTENT DNA content cell culture water: Sterile water containing NMT 0.005 USP Endotoxin Unit/mL DNA extraction buffer: Transfer 850 mL of Cell culture water to a sterile, 1-L graduated container. Dissolve 12.110 g of 2-amino-2-hydroxymethyl-1,3-propanediol, 3.802 g of edetate disodium, 23.380 g of sodium chloride, and 0.080 g of sodium dodecyl sulfate, with stirring. Adjust with 1 N hydrochloric acid or 1 N sodium hydroxide to a pH of 7.0. Dilute with Cell culture water to 1 L. Proteinase K solution: Prepare a solution of Tritirachium album proteinase K in 10 mM of 2-amino-2hydroxymethyl-1,3-propanediol, adjusted to a pH of 7.5, having an activity of 600 units/mL.15 Working DNA extraction buffer: Add 1.22 mL of Proteinase K solution to 38.78 mL of DNA extraction buffer. Dilution buffer: Transfer 850 mL of Cell culture water to a sterile, 1-L graduated container. Add 1.211 g of 2-amino-2hydroxymethyl-1,3-propanediol, 3.802 g of edetate disodium, and 5.844 g of sodium chloride, with stirring. Adjust with 1 N hydrochloric acid or 1 N sodium hydroxide to a pH of 7.0. Dilute with Cell culture water to 1 L. DPBS without Ca++, Mg++ solution: Prepare a solution containing 8.00 g of sodium chloride, 1.15 g of dibasic sodium phosphate (anhydrous), 0.20 g of potassium chloride, and 0.20 g/L of monobasic potassium phosphate. Calf thymus DNA solution: Prepare a solution containing 1 mg/L of calf thymus DNA in DPBS without Ca++, Mg++ solution, mixing thoroughly for 12 to 24 h at ambient temperature. Dilute the resulting solution with DPBS without Ca++, Mg++ solution to prepare a solution containing 50 g/mL of calf thymus DNA, mixing thoroughly on a vortex mixer for 10 min. Calf thymus DNA calibration solutions: Prepare four calibration solutions containing 5 g/mL, 10 g/mL, 15 g/mL, and 20 g/mL of calf thymus DNA, using Calf thymus DNA solution and diluting with DPBS without Ca ++, Mg++ solution. DNA staining solution: Prepare a solution containing 0.5 g of 2-(4-hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5bi-1H-benzimidazole trihydrochloride pentahydrate/mL of Dilution buffer. Store in low-actinic glassware. Analysis: Thaw Cryopreserved Human Fibroblast-Derived Dermal Substitute by placing the tissue, still in its ethyl vinyl acetate bag, in a water bath heated to between 34 and 37 for 2 to 3 min. The minimum amount of water in the water bath is 2 L/Cryopreserved Human Fibroblast-Derived Dermal Substitute unit. Cut three 11-mm 11-mm sections of Cryopreserved Human Fibroblast-Derived Dermal Substitute. To each of three microcentrifuge tubes add 1 mL of DPBS without Ca++, Mg++ solution. Immerse a single Cryopreserved Human Fibroblast-Derived Dermal Substitute 11-mm 11mm section into each microcentrifuge tube to remove the cryopreservative. Aspirate the DPBS without Ca++, Mg++ solution from each tube, and replace with 1 mL of Working DNA extraction buffer, making sure that each Cryopreserved Human Fibroblast-Derived Dermal Substitute is completely submerged in the extraction buffer. Incubate the samples in a 56 to 60 water bath for 4 to 18 h. Sonicate for 10 to 15 s using an ultrasonic cell disrupter to achieve complete
suitable Proteinase K solution can be obtained from Roche Diagnostics Corp., Roche Applied Sciences, P.O. Box 50414, 9115 Hague Rd., Indianapolis, IN 46250-0414.
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cellular disruption of the tissue and to mix the contents of the tube. Centrifuge the microcentrifuge tubes at 12,000 to 15,000 g to pellet non-DNA material. Transfer three 50-L aliquots of each sample supernatant to individual wells of a 96-well black plate suitable for performing fluorescent analysis. Transfer triplicate 50-L aliquots of each of the Calf thymus DNA calibration solutions to the 96-well plate, as well as a 50-L aliquot of DPBS working solution for the blank. Add 150 L of DNA staining solution to all wells containing the tissue samples, Calf thymus DNA calibration solutions, and the blank. Cover with aluminum foil, and place in a dark cabinet for 30 to 45 min at 15 to 30. Read the fluorescence of each well, using an excitation wavelength of 355 nm and an emission wavelength of 460 nm, blanking against the DPBS without Ca ++, Mg++ solution well. Plot the responses of the Calf thymus DNA calibration solutions versus concentration, in g/mL of calf thymus DNA, and calculate the regression line using the least-squares method. The test is considered valid if the %CV of the replicate values is less than 15%, the slope is between 4.48 and 6.27, the yintercept is between 2.04 and 3.65, and the square of the correlation coefficient is not less than 0.990. From the regression line so obtained, determine the amount of DNA, in g/11-mm 11-mm sample. Acceptance criteria: The amount of DNA of individual Cryopreserved Human Fibroblast-Derived Dermal Substitute 11-mm 11-mm section is between 6 and 15 g. TOTAL COLLAGEN CONTENT DPBS without Ca++, Mg++ solution: Proceed as directed for DNA content. DPBS with Ca++, Mg++ solution: Prepare a solution containing 8 g of sodium chloride, 1.15 g of dibasic sodium phosphate (anhydrous), 0.20 g of potassium chloride, 0.20 g of monobasic potassium phosphate, 0.10 g of magnesium chloride hexahydrate, and 0.10 g/L of calcium chloride (anhydrous). Collagenase extraction solution: Prepare a solution of Clostridium histolyticum collagenase, type 2, in DPBS with Ca++, Mg++ solution having an activity of at least 250 units/mL. 2% acetic acid solution: Mix 10 mL of acetic acid with 490 mL of water. Collagen standard stock solution: Prepare a solution having a concentration of 2 mg of type I collagen/mL of 2% acetic acid solution. Collagen calibration standards: Cut polyglactin mesh16 into seventeen 11-mm 11-mm squares, and place one square into 17 individual wells of a 24-well plate. Each well of the 24-well plate has a surface area of 220 mm2 and a volume of 3.5 mL. In quadruplicate, prepare wells containing 0.050 mg, 0.100 mg, 0.200 mg, and 0.400 mg of collagen by adding 25 mL, 50 mL, 100 mL, and 200 mL, respectively, of the Collagen standard stock solution. The remaining well to which no Collagen standard stock solution has been added is used as the blank. Allow the wells to air dry. Sirius red solution: Dissolve 0.5 g of Direct Red 80 in 500 mL of saturated picric acid. 1% (p-tert-Octylphenoxy) polyethoxyethanol solution: 10 mL of (p-tert-octylphenoxy) polyethoxyethanol in 990 mL of DPBS with Ca++, Mg++ solution. Analysis: Thaw Cryopreserved Human Fibroblast-Derived Dermal Substitute by placing the tissue, still in its ethyl vinyl acetate bag, in a water bath heated to between 34 and 37 for 2 to 3 min. The minimum amount of water in the water bath is 2 L/Cryopreserved Human Fibroblast-Derived Dermal Substitute unit. Cut three 11-mm 11-mm sections of Cryopreserved Human Fibroblast-Derived Dermal Substitute. Place each test section into separate wells of a 24-well plate.
16A suitable polyglactin mesh can be obtained from Ethicon Co., Johnson & Johnson Corp., 425 Hoes Ln., P.O. Box 6800, Piscataway, NJ 08855.
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Add 200 L of 1% (p-tert-Octylphenoxy) polyethoxyethanol solution to each sample. Shake on a rotating platform shaker at 100 to 150 rpm for 60 to 70 min at room temperature. Aspirate off the 1% (p-tert-Octylphenoxy) polyethoxyethanol solution, and rinse three times with 2 mL of DPBS without Ca++, Mg++ solution. Transfer each of the collagen standards to individual wells of the 24-well plate. Add 0.5 mL of Sirius red solution to each test sample and collagen standards. Shake on a rotating platform shaker at 100 to 150 rpm for 60 min at room temperature. Aspirate off the Sirius red solution from each well. Rinse each well twice with 2 mL of DPBS without Ca++, Mg++ solution. Add an additional 2 mL of DPBS without Ca++, Mg++ solution to each well, and allow to stand for 2 min. Aspirate off the DPBS without Ca++, Mg++ solution, and rinse twice more with 2 mL of DPBS without Ca++, Mg++ solution. Aspirate off all traces of DPBS without Ca++, Mg++ solution. Add 0.5 mL of Collagenase extraction solution to each well containing the Collagen calibration standards. Add 2 mL of Collagenase extraction solution to each well containing test samples. Rotate the plate on an orbital rotator at 150 rpm for 90 min at 37. Transfer 200 L from each well, and transfer to a suitable 96-well flatbottom plate. Read the absorbance of each aliquot at 540 nm. Dilute the Cryopreserved Human Fibroblast-Derived Dermal Substitute sample preparation further with DPBS without Ca++, Mg++ solution if the absorbance is greater than the absorbance of the highest of the Collagen calibration standards. Plot the responses of the Collagen calibration standards versus the amount, in mg of collagen, and calculate the regression line using the least-squares method. The test is considered valid if the slope is between 3.00 and 5.00 and the square of the correlation coefficient is greater than or equal to 0.950. Determine the collagen content, in mg, of an 11-mm x 11mm section of Dermal Substitute from the regression line as follows: Result = D A SCSR
USP 32
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Cryopreserved Human FibroblastDerived Dermal Substitute is aseptically packaged and supplied frozen in a clear ethyl vinyl acetate bag containing one piece of approximately 5 cm 7.5 cm for a single application. The solution within the bag is a saline-based cryoprotectant supplemented with 10% dimethyl sulfoxide and bovine serum to facilitate long-term storage. An aluminumplastic foil envelope surrounds the bag for its protection. Cryopreserved Human Fibroblast-Derived Dermal Substitute should be stored at 75 10 for no longer than 6 months. LABELING: The label indicates the dimensions of the Cryopreserved Human Fibroblast-Derived Dermal Substitute material enclosed. It contains the expiry date, the required storage conditions, and the lot number. The label cautions that Cryopreserved Human Fibroblast-Derived Dermal Substitute is not to be used if the package shows signs of damage. Additional labeling requirements include instructions on the proper thawing and handling of Cryopreserved Human Fibroblast-Derived Dermal Substitute, the timeframe for use after package opening, and a statement that cytotoxic agents are not to be used. USP REFERENCE STANDARDS 11 USP Endotoxin RS USP AUTHENTIC VISUAL REFERENCES 11 USP Cryopreserved Human Fibroblast-Derived Dermal Substitute Reference Photomicrographs [NOTEThese three photomicrographs represent examples of passing units. They are specified to assist in ascertaining histological quality.]
Desflurane
(Comment on this Monograph)id=m22832=Desflurane=DMonos.pdf)
= dilution factor (normally 4, unless the sample is further diluted) A = absorbance at 540 nm = slope of the regression line of the standards SCSR calculated above The test is considered valid if the slope of the regression line is between 3.00 and 5.00 and has a square of the correlation coefficient greater than 0.950. Acceptance criteria: The amount of collagen in individual Cryopreserved Human Fibroblast-Derived Dermal Substitute 11-mm 11-mm samples is between 0.40 and 2 mg. STERILITY TESTS 71: Meets the requirements Sample solution: Thaw Cryopreserved Human FibroblastDerived Dermal Substitute by placing the tissue, still in its ethyl vinyl acetate bag, in a water bath heated to a temperature of 34 to 37 for 2 to 3 min. The minimum amount of water in the water bath is 2 L/Cryopreserved Human Fibroblast-Derived Dermal Substitute unit. Analysis: Perform the test on 20 mL of the cryopreservative. BACTERIAL ENDOTOXINS TEST 85 Sample: Thaw Cryopreserved Human Fibroblast-Derived Dermal Substitute by placing the tissue, still in its ethyl vinyl acetate bag, in a water bath heated to a temperature of 34 to 37 for 2 to 3 min. The minimum amount of water in the water bath is 2 L/Cryopreserved Human Fibroblast-Derived Dermal Substitute unit. Remove the unit from the ethyl vinyl acetate bag, and immerse in 25 mL of LAL Reagent Water. Analysis: Extract for 60 min at 37 with shaking on an orbital shaker set at 125 revolutions/min. Remove a 4-mL aliquot of the extract for testing. Acceptance criteria: It contains NMT 0.5 USP Endotoxin Unit/mL.
C3H2F6O Ethane, 2-(difluoromethoxy)-1,1,1,2-tetrafluoro-, ()-; ()-2-Difluoromethyl 1,2,2,2-tetrafluoroethyl ether [57041-67-5].
168.04
DEFINITION Desflurane contains NLT 98.0% and NMT 102.0% of C3H2F6O. IDENTIFICATION The IR absorption spectrum of it using a gas cell exhibits maxima only at the same wavelengths as that of a similar preparation of USP Desflurane RS. ASSAY PROCEDURE Internal standard solution: 20 L/mL of USP Halothane RS in p-xylene Standard solution: Transfer 1.0 mL of Internal standard solution to a 2.0-mL septum-capped vial, cap, seal, and weigh accurately. Using a cold syringe, inject 25 L of USP Desflurane RS, previously cooled to 05, into the vial. Allow the vial to come to ambient temperature, weigh it, and calculate the quantity, in mg, of USP Desflurane RS added. Sample solution: Transfer 1.0 mL of Internal standard solution to a 2.0-mL septum-capped vial, cap, seal, and
USP 32
weigh accurately. Using a cold syringe, inject about 25 L of Desflurane, previously cooled to 0 to 5, into the vial. Allow the vial to come to ambient temperature, weigh it, and calculate the quantity, in mg, of Desflurane added. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2.4-mm 3.7-m stainless steel column coated with polytef and packed with 10% phase G31 and 15% phase G18 on 80- to 100-mesh support S1A Carrier gas: Helium Flow rate: 24 mL/min Temperature: See temperature table below.
Time Injector Detector Column 0 2.5 6.5 9.5 10.7 14.7 Temperature () 200 250 80 80 88 88 175 175
Official Monographs / Desflurane 27

Sample solution: Weigh a stoppered stock bottle containing a quantity of Desflurane at ambient temperature, and then cool it in powdered dry ice. Using a cold syringe, transfer 57 mL of Desflurane from the cold bottle to a separator containing 20 mL of Diluent A. Allow the stock bottle containing the remaining Desflurane to come to ambient temperature, weigh it, and calculate the quantity, in g, of Desflurane taken for the test. Allow the Desflurane in the separator to evaporate, and with the aid of a few mL of Diluent A, transfer the acid solution to a clean, dry beaker. Add 1 mL of hydrochloric acid to the solution in the beaker, and reduce the volume to 8 mL by evaporating on a hot plate. Transfer this solution to a 10-mL volumetric flask, and dilute with Diluent B to volume. Spectrometric conditions Mode: Atomic absorption spectrophotometer (see Spectrophotometry and Light-Scattering 851) Analytical wavelength: Antimony emission line at 217.6 nm Lamp: Antimony hollow-cathode Flame: Airacetylene Blank: Diluent B Analysis Samples: Standard solutions and Sample solution Calculation: Plot the absorbances of the Standard solutions versus concentration (g/mL) of antimony, and draw the straight line best fitting the three plotted points. From the graph so obtained, determine the concentration of antimony in the Sample solution. Calculate the quantity, in g/g, of antimony in the portion of Desflurane taken: Result =(C/W) V = concentration of antimony in the Sample solution (g/mL) W = weight of Desflurane taken to prepare the Sample solution (g) V = volume of Sample solution Acceptance criteria: NMT 3 g/g LIMIT OF FLUORIDE [NOTEStore all solutions except Solution A in plastic containers.] Solution A: 57 mL of glacial acetic acid, 58 g of sodium chloride, and 4 g of (1,2-cyclohexylenedinitrilo)tetraacetic acid in 500 mL of water. Adjust with 5 N sodium hydroxide to a pH of 5.25 0.25, and dilute with water to 1000 mL. Standard stock solution: Quantitatively dissolve a quantity of USP Sodium Fluoride RS in water to obtain a solution containing 2210 g/mL. Each mL of this solution contains 1000 g of fluoride/mL. Standard solutions: Dilute volumes of Standard stock solution with Solution A to obtain solutions having concentrations of 0.1, 0.3, 0.5, 1.0, 3.0, and 5.0 g of fluoride/mL. Sample solution: Transfer 10.0 mL of Desflurane to a 60mL separator, add 10.0 mL of water, shake for 1 min, and allow the layers to separate. Drain the lower organic layer and a small portion of the aqueous layer into a beaker, and discard. Transfer 5.0 mL of the aqueous phase remaining in the separator to a plastic cup, and add 5.0 mL of Solution A. [NOTETransfer the aqueous phase remaining in the separator to a second plastic cup, and reserve for the pH determination.] Analysis Samples: Standard solutions and Sample solution Concomitantly measure the potentials (see pH 791), in mV, of the Samples with a pH meter capable of a minimum reproducibility of 0.2 mV and equipped with C
Injection size: 1 L System suitability Sample: Standard solution [NOTEThe relative retention times for desflurane and halothane are 1.0 and 2.8, respectively.] Suitability requirements Resolution: NLT 8 between desflurane and halothane Relative standard deviation: NMT 1.0% for the ratios of the desflurane peak response to the halothane peak response obtained for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C3H2F6O in the portion taken: Result = (RU/RS) (WS/WU) 100 = ratio of the peak area response of desflurane to that of halothane from the Sample solution = ratio of the peak area response of desflurane to RS that of halothane from the Standard solution = weight of USP Desflurane RS used to prepare the WS Standard solution (mg) = weight of Desflurane used to prepare the Sample WU solution (mg) Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities LIMIT OF NONVOLATILE RESIDUE: Transfer 10.0 mL of it to an evaporating dish, evaporate with a stream of nitrogen to dryness, and dry the residue at 50 for 2 h: the weight of the residue does not exceed 7.5 mg (0.075%). LIMIT OF ANTIMONY Diluent A: Nitric acid and water (1:1) Diluent B: Nitric acid and hydrochloric acid (9:1) Standard solutions: Transfer 0.1 mL (234 mg) of antimony pentachloride to a 50-mL volumetric flask, dilute with Diluent B to volume, and mix. This stock solution contains about 1906 g of antimony/mL. Dilute a portion of this solution quantitatively and stepwise with Diluent B to obtain Standard solutions containing 2.5, 5.0, and 10.0 g of antimony/mL. RU
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PS
USP 32
= percentage of the respective impurity in the Standard solution rU = peak response of the respective impurity from the Sample solution = peak response of the respective impurity peak rS from the Standard solution Acceptance criteria: See Impurity Table.
Impurity Table Relative Retention Time 2.6 3.7 0.86 1.0 6.6 1.7 0.7 Relative Response Factor Acceptance Criteria, NMT (%) 1.0 0.1 0.1 0.05 0.025 0.01 0.01
a fluoride-specific ion-indicating electrode and a calomel reference electrode. [NOTEWhen taking measurements, immerse the electrodes in the solution, stir with a polytef-coated stirring bar and a magnetic stirrer having an insulated top until equilibrium is attained (about 12 min), and record the potential. Rinse the electrodes with Solution A, and dry, taking care to avoid damaging the crystal of the specific-ion electrode.] Plot the logarithms of the fluoride concentrations (g/mL) of the Standard solutions versus potential, in mV. From the measured potential of the Sample solution and the standard response line, determine the concentration, C (g/mL), of fluoride in the Sample solution. Multiply C by 0.0002 to obtain the percentage of fluoride in the Desflurane taken. Acceptance criteria: NMT 0.001% Organic Impurities PROCEDURE Standard solution: Insert a rubber-sleeve stopper into a 2mL vial, weigh accurately, and partially evacuate the vial. Using a cold syringe, inject about 680 L of USP Desflurane RS, previously cooled to 05, into the vial. Allow the vial to come to ambient temperature, weigh it, and calculate the quantity (mg) of USP Desflurane RS added. Using a syringe, inject about 7 L of USP Desflurane Related Compound A RS into the same vial, weigh, and calculate the quantity (mg) of USP Desflurane Related Compound A RS added. Similarly and in turn, add about 7.2 L of trichlorofluoromethane, 7.4 L of dichlorofluoromethane, 7.6 L of methylene chloride, 6.8 L of chloroform, 6.3 L of trichlorotrifluoroethane, and 6.7 L of isoflurane, weighing the vial after each addition and calculating the quantity (mg) of each addition. Calculate the percentage of each impurity in the Standard solution taken: Result = 100 Wi/(WD + WT) = quantity of the respective impurity added during the preparation of the Standard solution (mg) = quantity of USP Desflurane RS in the Standard WD solution (mg) = sum of all the impurities added during the WT preparation of the Standard solution (mg) Sample solution: Desflurane Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2.4 -mm 6.1-m stainless steel column packed with 25% phase G16 on 80- to 100-mesh support S1A Carrier gas: Helium Flow rate: 20 mL/min Temperature Column: 75 Injection port: 200 Detector: 250 Injection size: 1 L Run time: 40 min System suitability Sample: Standard solution Suitability requirements Resolution: NLT 1.2 between trichlorofluoromethane and desflurane related compound A Tailing factor: NMT 1.5 for the desflurane related compound A peak Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity taken: Result = 2 PS (rU/rS) Wi
Name Isoflurane Methylene chloride Desflurane related compound A Desflurane Chloroform Trichlorotrifluoroethane Dichlorofluoromethane Trichlorofluoromethane
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at 25; excursions are permitted between 15 and 30. Replace the cap securely after each use. USP REFERENCE STANDARDS 11 USP Desflurane RS USP Desflurane Related Compound A RS USP Halothane RS USP Sodium Fluoride RS
Desipramine Hydrochloride
(Comment on this Monograph)id=m22840=Desipramine Hydrochloride=D-Monos.pdf)
302.84 C18H22N2 HCl 5H-Dibenz[b,f]azepine-5-propanamine, 10,11-dihydroN-methyl-, monohydrochloride; 10,11-Dihydro-5-[3-(methylamino)propyl]-5H-dibenz[b,f]azepine monohydrochloride [58-28-6]. DEFINITION Desipramine Hydrochloride, dried in vacuum at 105 for 2 h, contains NLT 98.0% and NMT 100.5% of C18H22N2 HCl. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 251 nm Sample solution: 30 g/mL in 0.1 N hydrochloric acid Acceptance criteria: Absorptivities, calculated on the dried basis, do not differ by more than 2.0%. C. Add 5 mg to 2 mL of nitric acid on a spot plate. Acceptance criteria: An intense blue color is produced. D. IDENTIFICATION TESTSGENERAL, Chloride 191: 50 mg/mL in alcohol
USP 32
ASSAY PROCEDURE Sample: 0.6 g of previously dried and weighed Desipramine Hydrochloride Analysis: Dissolve Sample in 100 mL of glacial acetic acid, add 10 mL of mercuric acetate TS. Titrate with 0.1 N perchloric acid VS, determining the endpoint potentiometrically, using a calomel-glass electrode system. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 30.29 mg of C18H22N2 HCl. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 10 ppm Organic Impurities PROCEDURE: LIMIT OF IMINODIBENZYL Standard stock solution: 50 g/mL of USP Iminodibenzyl RS in alcohol Standard solution: 5 g/mL of USP Iminodibenzyl RS from Standard stock solution and a mixture of of hydrochloric acid and alcohol (1:1) [NOTEUse a lowactinic volumetric flask.] Sample solution: 5 mg/mL of Desipramine Hydrochloride in a mixture of hydrochloric acid and alcohol (1:1) [NOTEUse a low-actinic volumetric flask.] Spectrometric conditions Mode: UV-Vis Analytical wavelength: 565 nm Cell: 1 cm Blank: Hydrochloric acid and alcohol (1:1) Analysis Samples: Standard solution, Sample solution, and Blank To the two flasks containing the Standard solution and the Sample solution, and to a third flask containing 10 mL of Blank, add slowly 5 mL of a 0.4% solution of furfural in alcohol, then add 5 mL of hydrochloric acid, and allow the flasks to stand in a constant-temperature bath at 25 for 3 h. Dilute each flask with a mixture of equal volumes of hydrochloric acid and alcohol to volume. Determine the absorbances of the solutions, using the Blank to set the instrument. Acceptance criteria: The absorbance of the solution obtained from the Sample solution is NMT that obtained from the Standard solution (0.1%). SPECIFIC TESTS LOSS ON DRYING 731: Dry it in vacuum at 105 for 2 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Desipramine Hydrochloride RS USP Iminodibenzyl RS
Official Monographs / Desipramine 29

powder with 15 mL of chloroform. Pass the chloroform extract through paper into a wide-mouth test tube, and evaporate the filtrate to 3 mL. Carefully add ether until the liquid becomes turbid, heat cautiously to produce a clear solution, then cool, and allow to stand. Collect the crystals, wash with ether, and dry in vacuum at 80 for 30 min. ASSAY PROCEDURE Standard solution: 25 g/mL of USP Desipramine Hydrochloride RS in 0.1 N hydrochloric acid Sample solution: Finely powder a number of Tablets, equivalent to 0.5 g of desipramine hydrochloride, and transfer the powder to a 500-mL volumetric flask with the aid of 100 mL of 0.1 N hydrochloric acid. Add 150 mL of the 0.1 N acid to the mixture, insert the stopper, and shake by mechanical means for 30 min. Dilute with 0.1 N hydrochloric acid to volume and filter discarding the first 10 mL of the filtrate. Transfer 5.0 mL of the subsequent filtrate to a 200-mL volumetric flask, and dilute with 0.1 N hydrochloric acid to volume. Spectrometric conditions Mode: UV Analytical wavelength: 255 nm Cell: 1 cm Blank: Cyclohexane Analysis Samples: Standard solution and Sample solution Transfer 15.0 mL each of the Standard solution and the Sample solution to separate glass-stoppered, 50-mL centrifuge tubes, and to each tube add 2 mL of 2.5 N sodium hydroxide and 15.0 mL of cyclohexane suitable for use in UV spectrophotometry. Insert the stoppers, shake the tubes by mechanical means for 30 min, and centrifuge at 1500 rpm until the clear phases separate (5 to 10 min). Determine the absorbances of the cyclohexane extracts, using the Blank. Calculate the percentage of C18H22N2 HCl in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Desipramine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 95.0%105.0% AU AS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: 0.1 N hydrochloric acid; 900 mL Apparatus 2: 50 rpm Time: 60 min Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to that of the Standard solution Standard solution: USP Desipramine Hydrochloride RS in Medium Spectrometric conditions Mode: UV Analytical wavelength: 251 nm Analysis Samples: Standard solution and Sample solution Tolerances: NLT 75% (Q) of the labeled amount of C18H22N2 HCl
Desipramine Hydrochloride Tablets

(Comment on this Monograph)id=m22860=Desipramine Hydrochloride Tablets=D-Monos.pdf) DEFINITION Desipramine Hydrochloride Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of C18H22N2 HCl. IDENTIFICATION INFRARED ABSORPTION 197M Sample: Finely powder a number of Tablets, equivalent to 350 mg of desipramine hydrochloride, and triturate the
30
Desipramine / Official Monographs
USP 32
Adsorbent: 0.25-mm layer of chromatographic silica gel. Application volume: 5 L Developing solvent system: Methylene chloride, methanol, and water (130:36:3) Spray reagent: Dilute perchloric acid (1 in 20) Analysis Samples: Standard solution and Sample solution Develop chromatogram until the solvent has moved threefourths of the length of the plate. Locate the spots on the plate by lightly spraying with Spray reagent and heating at about 100 for 3 min. Cool, and examine under UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Standard solution: 0.2 mg/mL of USP Deslanoside RS in alcohol Sample solution: 0.2 mg/mL of Deslanoside in alcohol Spectrometric conditions Mode: UV-Vis Analytical wavelength: 590 nm Cell: 1 cm Blank: Alcohol Analysis Samples: Standard solution, Sample solution, and Blank Transfer 3.0 mL each of the Standard solution, the Sample solution, and the Blank, to separate 25-mL conical flasks. Evaporate each with gentle warming and with the aid of a current of air just to dryness, and cool in a vacuum desiccator for 30 min. Add 15.0 mL of acid-ferric chloride TS to each flask, mix by swirling, and allow the mixtures to stand protected from light, swirling frequently, at a temperature not exceeding 30, for 15 min. Pass each through separate fine glass wool filters. Concomitantly determine the absorbances of the solutions. Repeat the measurements at 2-min intervals until maximum absorbance readings have been obtained. Calculate the percentage of C47H74O19 in the portion of Deslanoside taken: Result = (CS/CU) (AU/AS) 100 = concentration of USP Deslanoside RS in the Standard solution (mg/mL) = nominal concentration of deslanoside in the CU Sample solution (mg/mL) AU = maximum absorbances of the solutions from the Sample solution = maximum absorbances of the solutions from the AS Standard solution Acceptance criteria: 95.0%103.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: CS
UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Procedure for content uniformity Sample solution: 25 g/mL of desipramine hydrochloride in 0.1 N hydrochloric acid [NOTETransfer 1 finely powdered Tablet to a 100-mL volumetric flask, add 50 mL of 0.1 N hydrochloric acid, and shake by mechanical means for 15 min. Dilute with 0.1 N hydrochloric acid to volume, and filter, discarding the first 20 mL of the filtrate. Dilute a portion of the subsequent filtrate quantitatively and stepwise with 0.1 N hydrochloric acid.] Standard solution: 25 g/mL of USP Desipramine Hydrochloride RS in 0.1 N hydrochloric acid Spectrometric conditions Cell: 1 cm Analytical wavelength: 251 nm Blank: 0.1 N hydrochloric acid Analysis Samples: Sample solution, Standard solution, and Blank Calculate the percentage of C18H22N2 HCl in the Tablet taken: Result = (AU/AS) (CS/CU) 100 AU AS CS CU = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Desipramine Hydrochloride RS in the Standard solution (g/mL) = nominal concentration of desipramine hydrochloride in the Sample solution (g/mL)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Desipramine Hydrochloride RS
Deslanoside
(Comment on this Monograph)id=m22890=Deslanoside=DMonos.pdf)
943.08 C47H74O19 Card-20(22)-enolide, 3-[(O--D-glucopyranosyl-(14)-O-2,6dideoxy--D-ribo-hexopyranosyl-(14)-O-2,6-dideoxy--D-ribohexopyranosyl-(14)-2,6-dideoxy--D-ribohexopyranosyl)oxy]-12,14-dihydroxy-, (3,5,12)-; Deacetyllanatoside C [17598-65-1]. DEFINITION Deslanoside contains NLT 95.0% and NMT 103.0% of C47H74O19, calculated on the dried basis. [CAUTIONHandle Deslanoside with exceptional care, since it is highly potent.] IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 4 mg/mL of USP Deslanoside RS in methanol Sample solution: 4 mg/mL in methanol Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.)
NMT 0.2%
SPECIFIC TESTS SPECIFIC ROTATION 781S: +7.0 to +8.5 Sample solution: 20 mg/mL, in anhydrous pyridine. LOSS ON DRYING 731: Dry 0.5 g in vacuum at 100 to constant weight: it loses NMT 5.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Deslanoside RS
USP 32
Official Monographs / Desmopressin 31

= maximum absorbance of the Standard solution = concentration of USP Deslanoside RS in the Standard solution (g/mL) CU = nominal concentration of deslanoside in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% SPECIFIC TESTS PH 791: 5.57.0 OTHER REQUIREMENTS: Injections 1. AS CS
Deslanoside Injection
(Comment on this Monograph)id=m22900=Deslanoside Injection=D-Monos.pdf) DEFINITION Deslanoside Injection is a sterile solution of Deslanoside in a suitable solvent. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C47H74O19. It may contain Glycerin. IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 4 mg/mL of USP Deslanoside RS in methanol Sample solution: Transfer a volume of Injection, equivalent to 2 mg of deslanoside, to a small separator, and extract with 25 mL of a mixture of chloroform and alcohol (7:3). Transfer the extract to a 10-mL conical flask, evaporate on a steam bath to dryness, and dissolve the residue in 500 L of methanol. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume: 5 L Developing solvent system: Methylene chloride, methanol, and water (130:36:3) Spray reagent: Dilute perchloric acid (1 in 20) Analysis: Locate the spots on the plate by lightly spraying with spray reagent and heating at about 100 for 3 min. Cool, and examine under UV light: the RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Standard solution: 200 g/mL of USP Deslanoside RS in alcohol Sample solution: Transfer a volume of Injection, nominally equivalent to 600 g of deslanoside, to a separator, and add 50 mL of water and 1 mL of 2 N sulfuric acid. Extract four 30-mL portions of a mixture of chloroform and n-propyl alcohol (5:1), washing each portion in a s separator containing 5 mL of water and filtering through cotton that previously has been moistened with chloroform. Combine the extracts, and evaporate on a steam bath, with the aid of a current of air, just to dryness. Transfer the residue, with the aid of a small volume of the chloroformn-propyl alcohol mixture, to a 25-mL conical flask. Spectrometric conditions Mode: UV-Vis Analytical wavelength: 590 nm Cell: 1 cm Blank: Alcohol Analysis Samples: Standard solution, Sample solution, and Blank Transfer 3.0 mL each of the Samples to separate 25-mL conical flasks. Evaporate each with gentle warming and with the aid of a current of air just to dryness, and cool in a vacuum desiccator for 30 min. Add 15.0 mL of acidferric chloride TS to each flask, mix by swirling, and allow the mixtures to stand protected from light, swirling frequently, at a temperature not exceeding 30, for 15 min. Pass each through separate fine glass wool filters. Concomitantly determine the absorbances of the solutions. Repeat the measurements at 2-min intervals until maximum absorbance readings have been obtained. Calculate the quantity, in g, of C47H74O19 in each mL portion of the Injection taken: Result = (AU/AS) (CS/CU) 100 AU = maximum absorbance of the Sample solution
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose containers, preferably of Type I glass. USP REFERENCE STANDARDS 11 USP Deslanoside RS
Desmopressin Acetate
(Comment on this Monograph)id=m22930=Desmopressin Acetate=D-Monos.pdf)
1129.27 C48H68N14O14S2C48H68N14O14S2 xH2O (anhydrous) [62288-83-9]; Vasopressin, 1-(3-mercaptopropanoic acid)-8-D-arginine-, monoacetate (salt); 1-(3-Mercaptopropionic acid)-8-D-arginine-vasopressin monoacetate (salt); Trihydrate 1183.31 [62357-86-2]. DEFINITION Desmopressin Acetate is a synthetic octapeptide hormone having the property of antidiuresis. It is a synthetic analog of vasopressin. It contains NLT 95.0% and NMT 105.0% of desmopressin (C46H64N14O12S2), calculated on the anhydrous, acetic acid-free basis. IDENTIFICATION A. MASS SPECTRAL ANALYSIS Diluent: Methanol and water (1:1) Standard solution: 5 g/mL of USP Desmopressin Acetate RS in Diluent Sample solution: 5 g/mL of Desmopressin Acetate in Diluent [NOTEThe final concentration of the Standard solution and the Sample solution can be adjusted depending on the sensitivity of the mass spectrometer used in the testing.] Spectrometric conditions (See Mass Spectrometry 736.) Mode: LC/MS, equipped with an electrospray interface, positive ion mode, infusion system, and MS/MS capability Introduction: Electrospray Ion mode: Positive Injection type: Infusion system Infusion rate: 5 L/min Analysis Samples: Standard solution and Sample solution Obtain optimized MS and MS/MS spectra of the peak with mass-to-charge ratio 1069. Acceptance criteria: For MS spectra, the major peak with mass-to-charge ratio of 1069 should be observed. For
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Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Column temperature: 30 Flow rate: 1 mL/min Injection size: 200 L System suitability [NOTEThe desmopressin peak elutes before the oxytocin peak.] Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 1.5 between desmopressin and oxytocin Tailing factor: NMT 2.0 Relative standard deviation: NMT 5.0%, Standard solution Analysis Sample: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Desmopressin Acetate taken: Result = (rU/rS) (CS/CU) 100 = peak response for the relevant impurity from the Sample solution = peak response for desmopressin from the rS Standard solution = concentration of USP Desmopressin Acetate RS CS in the Standard solution, calculated on the anhydrous, acetic acid-free basis (mg/mL) = concentration of Desmopressin Acetate in the CU Sample solution, calculated on the anhydrous, acetic acid-free basis (mg/mL) Acceptance criteria Individual impurities: NMT 0.5% Total impurities: NMT 1.5% PROCEDURE 2: LIMIT OF ACETIC ACID Internal standard solution: Transfer 16 mL of hydrochloric acid to a 1000-mL volumetric flask containing 500 mL of water. Add 0.5 mL of propionic acid and dilute with acetonitrile to volume. Standard solution: Transfer 1.049 g of acetic acid to a 100-mL volumetric flask and dilute with Internal standard solution to volume. Transfer 2.5 mL of this resulting solution to a 50-mL volumetric flask and dilute with Internal standard solution to volume. Sample solution: Dissolve 5 mg of Desmopressin Acetate in 0.5 mL of Internal standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: FID Column: 0.32-mm 30-m fused silica; 0.25-m coating of G35 Temperature Column: 120 Injector: 250 Detector: 250 Carrier gas: Helium Flow rate: 3 mL/min [NOTESplit ratio (20:3)] Injection size: 1.0 L System suitability Sample: Standard solution (six replicate injections) [NOTEAcetic acid elutes before propionic acid.] Suitability requirements Resolution: NLT 5.0 between acetic acid and propionic acid Tailing factor: NMT 3.0 Relative standard deviation: NMT 15% for the peak area ratio of acetic acid to propionic acid rU
MS/MS spectra, product ions at mass-to-charge ratios of about 641, 742, and 995 are present. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Solution A: Dissolve 3.4 g of monobasic potassium phosphate and 2.0 g of sodium 1-heptanesulfonic acid in 1000 mL of water. Adjust the pH to 4.50 0.05 with phosphoric acid or sodium hydroxide, as needed. Mobile phase: Acetonitrile and Solution A (220:780) [NOTEThe retention time of desmopressin is very sensitive to the composition of the Mobile phase.] Standard solution: 20 g/mL of USP Desmopressin Acetate RS in Mobile phase Sample solution: 20 g/mL of Desmopressin Acetate in Mobile phase System suitability solution: Transfer 1 mg of oxytocin to a 50-mL volumetric flask. Dissolve in and dilute with Mobile phase to volume. Transfer 5.0 mL of this solution and 5.0 mL of the Sample solution to a 100-mL volumetric flask and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 220 nm Column: 4.6-mm 25-cm; 5-m packing L1 Column temperature: 30 Flow rate: 1 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution [NOTEThe desmopressin peak elutes before the oxytocin peak.] Suitability requirements Resolution: NLT 1.5, between desmopressin and oxytocin peaks Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% for the desmopressin peak area Analysis Samples: Standard solution and Sample solution Calculate the percentage of C46H64N14O12S2 in the portion of Desmopressin Acetate taken: Result = (rU/rS) (CS/CU) 100 = peak response for desmopressin from the Sample solution = peak response for desmopressin from the rS Standard solution = concentration of USP Desmopressin Acetate RS CS in the Standard solution, calculated on the anhydrous, acetic acid-free basis (mg/mL) = concentration of Desmopressin Acetate, CU calculated on the anhydrous, acetic acid-free basis, in the Sample solution (mg/mL) Acceptance criteria: 95.0%105.0% rU IMPURITIES Organic Impurities PROCEDURE 1 Solution A, Mobile phase, and System suitability solution: Proceed as directed in the Assay. Standard solution: 1 g/mL of USP Desmopressin Acetate RS in Mobile phase Sample solution: 200 g/mL of Desmopressin Acetate in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.)
USP 32
Analysis Samples: Standard solution and Sample solution Calculate the percentage of acetic acid in the portion of Desmopressin Acetate taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of acetic acid to the internal standard peak from the Sample solution = peak response ratio of acetic acid to the internal RS standard peak from the Standard solution = concentration of acetic acid in the Standard CS solution (mg/mL) = concentration of Desmopressin Acetate in the CU Sample solution (mg/mL) Acceptance criteria: 3%8% RU SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIC MICROORGANISMS 62: The total aerobic microbial count is NMT 100 cfu/g. OPTICAL ROTATION, Specific Rotation 781S: 72 to 82, calculated on the anhydrous, acetic acid-free basis Sample solution: 5 mg/mL in diluted acetic acid WATER DETERMINATION, Method Ic 921: NMT 6.0% AMINO ACID CONTENT (See Biotechnology-Derived ArticlesAmino Acid Analysis 1052.) [NOTEThe following method is given for informational purposes; any validated amino acid analysis method can be used. The relative proportions of amino acids, however, must be met for any method used.] Solution A: Prepare a solution having final concentrations of 20 mM sodium acetate, 0.2% triethylamine, and 0.3% tetrahydrofuran. Solution B: Prepare a solution containing 20% 100 mM sodium acetate, 40% methanol, and 40% acetonitrile. Mobile phase: See the gradient table below.
Time (min) 0 17 18.10 24 25 35 Solution A (%) 100 40 0 0 100 100 Solution B (%) 0 60 100 100 0 0
Official Monographs / Desmopressin 33

2.50 mM of L-aspartic acid 2.50 mM of L-threonine 2.50 mM of L-serine 2.50 mM of L-glutamic acid 2.50 mM of L-proline 2.50 mM of L-alanine 2.50 mM of L-valine 2.50 mM of L-methionine 2.50 mM of L-isoleucine 2.50 mM of L-leucine 2.50 mM of L-tyrosine 2.50 mM of L-phenylalanine 1.25 mM of L-cystine Transfer a 1-mL aliquot of this solution to a suitable vial, and add 5 L each of Norvaline solution and DTDPA solution. Evaporate the aliquot to dryness, and add 300 L of Phenol solution. Mix, and alternate between purging the head space of the vial with nitrogen gas and reducing the pressure to 2 mm of mercury for a total of two purgevacuum cycles. Purge the sample with nitrogen gas one additional time, and reduce the pressure to 1.5 mm of mercury. Seal, and heat the sample at 110 for 24 h. Open the vial, and evaporate to dryness. Dissolve the residue in 115 L of Borate buffer, and add 5 L of Sarcosine solution. Centrifuge for 2 min, and transfer the supernatant to a clean vial. Remove a 6-L aliquot, and add 1 L of OPA reagent. Mix, and add 1 L of FMOC reagent. Mix, add 28 L of water, and mix again. Sample solution: Prepare a solution of Desmopressin Acetate in water having a concentration of 1.00 mg/mL. Add 5 L each of Norvaline solution and DTDPA solution to a 1-mL aliquot, and prepare as directed in Calibration solution, beginning with Evaporate the aliquot to dryness and add 300 L of Phenol solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 262 nm and 338 nm Column: 2.1-mm 20-cm; 5-m packing L1 Temperature Thermoregulated autosampler: 4 [NOTEThe autosampler must be able to add and mix reagents.] Column: 40 Flow rate: 0.45 mL/min Injection size: 6 L System suitability Samples: Calibration solution and Sample solution [NOTEThe order of elution of the amino acid derivatives is aspartic acid, glutamic acid, serine, histidine, glycine, threonine, cysteine (reduced cystine), alanine, arginine, tyrosine, valine, methionine, norvaline, phenylalanine, isoleucine, leucine, lysine, and proline.] Suitability requirements Resolution: NLT 1, between histidine and glycine; NLT 1, between alanine and arginine; and NLT 1, between valine and methionine, Calibration solution Peak response: The peak area of the norvaline peak in the Sample solution is NLT 80% and NMT 120% of that found in the Calibration solution. Relative standard deviation: NMT 15% for cysteine, lysine, and proline, and NMT 10% for all other amino acids from triplicate injections, Calibration solution Analysis Samples: Calibration solution and Sample solution Using the autosampler, separately remove equal volumes (about 6 L) of the Calibration solution and the Sample solution, and to each add 1 L of OPA reagent. Mix, and to each add 1 L of FMOC reagent. Mix, add 28 L of water to each, mix again, and inject the entire volume into the chromatograph. Record the area responses for
Borate buffer: Transfer 12.4 g of boric acid to a 500-mL volumetric flask, and suspend it in 300 mL of water. Add 100 mL of 1 N potassium hydroxide. Adjust with 1 N potassium hydroxide to a pH of 10.4, and dilute with water to volume. Store in a closed plastic container. Norvaline solution: 4 mM norvaline DTDPA solution: 20 mg/mL of dithiodipropionic acid in water Sarcosine solution: 4 mM sarcosine in water Phenol solution: 0.1% (w/v) phenol in 6 N hydrochloric acid OPA reagent: 10 mg/mL each of o-phthalaldehyde and 3mercaptopropionic acid in Borate buffer FMOC reagent: 2.5 mg/mL of 9fluorenylmethylchloroformate in acetonitrile Calibration solution: Prepare a mixture in which the final concentrations of amino acids are as follows: 2.50 mM glycine 2.50 mM of L-lysine 2.50 mM of L-histidine 2.50 mM of L-arginine
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solution with Solution A or Solution B, as directed for the Sample solution, to obtain a solution with a concentration of desmopressin equivalent to that in the Sample solution. Sample solution: If labeled content of desmopressin acetate is: 4 g/mL0.1 mg/mL: Use undiluted Injection Greater than 0.1 mg/mL without preservatives: Dilute 1000 L of Injection with 10 mL of Solution A. Greater than 0.1 mg/mL with preservatives: Dilute 1000 L of Injection with 10 mL of Solution B. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 215 nm Column: 4.6-mm 25-cm; 5-m L1 packing Flow rate: 1.5 mL/min Injection size: 100 L (50 L for System suitability) System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.4 Relative standard deviation: NMT 5.0% Analysis Samples: Standard solution and Sample solution [NOTERecord chromatograms NLT 2.5 times the retention time of desmopressin.] Calculate the percentage of C46H64N14O12S2 in the portion of Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of desmopressin in the Standard solution (mg/mL) CU = nominal concentration of desmopressin acetate in the Sample solution Acceptance criteria: 90.0%100.0% SPECIFIC TESTS PH 791: 3.56.0 BACTERIAL ENDOTOXINS TEST 85: Contains NMT 10 USP Endotoxin Unit/mg of desmopressin STERILITY TESTS, Test for Sterility of the Product to be Examined 71: Meets the requirements when tested as directed for Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements for small-volume injections. INJECTIONS, Volume in Container 1: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, protected from light, between 28. LABELING: Label it to state the potency, in mg, of desmopressin. USP REFERENCE STANDARDS 11 USP Desmopressin Acetate RS USP Endotoxin RS rU rS CS
the main peaks, and identify the amino acids. Using the Calibration solution as a standard, express the content of each amino acid in moles. With the content for arginine set to 1, calculate the relative proportions of the amino acids. Acceptance criteria: See Amino Acid Acceptance Table.
Amino Acid Acceptance Table Acceptance Criteria (mole ratio) Name Aspartic Acid Glutamic acid Proline Glycine Phenylalanine Tyrosine Cysteine Lysine Isoleucine Leucine All others NLT 0.95 0.95 0.95 0.95 0.95 0.7 0.3 Absent Absent Absent NMT 1.05 1.05 1.05 1.05 1.05 1.05 1.05 Absent Absent Absent NMT Trace
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers, preferably of Type I glass, protected from light, and store at a NMT 25, preferably 28. LABELING: Label it to state the potency, in mg, of desmopressin USP REFERENCE STANDARDS 11 USP Desmopressin Acetate RS
Desmopressin Acetate Injection

(Comment on this Monograph)id=m22937=Desmopressin Acetate Injection=D-Monos.pdf) DEFINITION Desmopressin Acetate Injection is a sterile solution of Desmopressin Acetate in a suitable diluent. It may contain suitable preservatives. It possesses, in each mL, an activity of NLT 90.0% and NMT 110.0% of the labeled amount of desmopressin (C46H64N14O12S2), calculated on the anhydrous, acetic acid-free basis. IDENTIFICATION The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Buffer: Dissolve 4.9 g of phosphoric acid in water and dilute with water to 1000 mL. Adjust with triethylamine to a pH of 3.5. Mobile phase: Acetonitrile and Buffer (33:167) Solution A: 9 mg/mL of sodium chloride in water. Adjust the pH to 3.55.0 with hydrochloric acid Solution B: 9 mg/mL of sodium chloride and 5 mg/mL of chlorobutanol in water. Adjust the pH to 3.55.0 with hydrochloric acid. [NOTEDissolve sodium chloride with water prior to adding chlorobutanol.] Standard solution: Prepare a solution containing 1 mg/mL of USP Desmopressin Acetate RS in water. Dilute this
Desogestrel and Ethinyl Estradiol Tablets

(Comment on this Monograph)id=m22965=Desogestrel and Ethinyl Estradiol Tablets=D-Monos.pdf) DEFINITION Desogestrel and Ethinyl Estradiol Tablets contain NLT 90.0% and NMT 110.0% of the labeled amounts of desogestrel (C22H30O) and ethinyl estradiol (C20H24O2).
USP 32
IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 0.15 mg/mL of USP Desogestrel RS and 0.03 mg/mL of USP Ethinyl Estradiol RS in ether Sample solution: Transfer a number of Tablets, equivalent to 1.5 mg desogestrel and 0.3 mg ethinyl estradiol, to a suitable container, add 50 mL of water, and sonicate until the Tablets disintegrate (if necessary, remove any coating with water before sonication). Place the sample in a separatory funnel, add 25 mL of ether, and shake well to extract the actives. Using a glass pipet, transfer the ether layer to a clean beaker, and evaporate to about 10 mL. Application volume: 30 L Developing solvent system: Chloroform and alcohol (96:4) Spray reagent: Methanol and sulfuric acid (1:1) Analysis: Proceed as directed in the chapter, and then airdry. Spray the plate with the Spray reagent, place in an oven at 105 for about 5 min, and examine the plate. Acceptance criteria: Meets the requirements B. The retention time of the major peaks of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and 20 mM potassium phosphate buffer, pH 6.0 (1:1) Diluent: Acetonitrile and water (1:1) Standard stock solution A: 0.3 mg/mL of USP Desogestrel RS in methanol Standard stock solution B: 0.3 mg/mL of USP Ethinyl Estradiol RS in methanol Standard solution: Dilute appropriate aliquots of Standard stock solution A and Standard stock solution B in methanol to obtain a solution having a known concentration of 0.6 g/mL of USP Desogestrel RS and 0.12 g/mL of USP Ethinyl Estradiol RS. Sample solution: Transfer 20 Tablets into a 200-mL volumetric flask. Add about 120 mL of Diluent, and shake for about 30 min. Dilute with Diluent to volume, and mix. Centrifuge a portion of the sample, and dilute with Diluent to obtain a solution containing nominally 0.6 g/mL of desogestrel. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detectors Desogestrel analysis: UV 210 nm Ethinyl estradiol analysis: Spectrofluorometric detector, excitation at 285 nm and emission at 310 nm Analytical column: 4.6-mm 15-cm; packing L11 Guard column: 4.6-mm 12.5-cm; packing L11 Flow rate: 2 mL/min Injection size: 200 L System suitability Sample: Standard solution [NOTEThe relative retention times for ethinyl estradiol and for desogestrel are 0.2 and 1.0, respectively.] Suitability requirements Tailing factor: NMT 2.0 for both ethinyl estradiol and desogestrel Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30O and C20H24O2 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU = peak response for the relevant analyte of the Sample solution rS
Official Monographs / Desogestrel 35

= peak response for the relevant analyte of the Standard solution = concentration of the appropriate Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of relevant analyte in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION 711 Test 1 Medium: 0.05% sodium lauryl sulfate with an assay content of NLT 95%; 500 mL Apparatus 2: 50 rpm Time: 30 min Mobile phase: Acetonitrile and 20 mM potassium phosphate buffer, pH 6.0 (1:1) Sample solution: Sample per Dissolution 711. Centrifuge a portion of the dissolution sample and use the clear supernatant. Standard stock solution A: 0.25 mg/mL of USP Desogestrel RS in methanol. Dilute 1.0 mL of this solution with Medium to 50.0 mL. Standard stock solution B: 0.25 mg/mL of USP Ethinyl Estradiol RS in methanol. Dilute 1.0 mL of this solution with Medium to 50.0 mL. Standard solution: Dilute portions of Standard stock solution A and Standard stock solution B with Medium to obtain a solution containing 0.3 g/mL of USP Desogestrel RS and 0.06 g/mL of USP Ethinyl Estradiol RS. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detectors Desogestrel analysis: UV 210 nm Ethinyl estradiol analysis: Spectrofluorometric detector, excitation at 285 nm and emission at 310 nm Analytical column: 4.6-mm 15-cm; packing L11 Guard column: 4.6-mm 12.5-cm; packing L11 Flow rate: 2 mL/min Injection size: 200 L System suitability Sample: Standard solution [NOTEThe relative retention times for ethinyl estradiol and for desogestrel are 0.2 and 1.0, respectively.] Suitability requirements Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30O and C20H24O2 dissolved: Result = (rU/rS) (CS/CU) 100 = peak response for the relevant analyte of the Sample solution = peak response for the relevant analyte of the rS Standard solution = concentration of the appropriate Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of relevant analyte in the CU Sample solution (mg/mL) Tolerances: NLT 80% (Q) of the labeled amount of C22H30O and C20H24O2 is dissolved. Test 2 Medium: 0.3% sodium lauryl sulfate; 500 mL Apparatus 2: 100 rpm Time: 30 min Sample solution: Sample per Dissolution 711. Centrifuge a portion of the dissolution sample and use the clear supernatant. rU
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ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water, (65:1:35), such that the retention time of desoximetasone is about 6 min Standard stock solution: 0.4 mg/mL of USP Desoximetasone RS in methanol Standard solution: 0.04 mg/mL of USP Desoximetasone RS from Standard stock solution, and a (1:1) mixture of methanol and acetonitrile [10:90] Sample stock solution: 0.4 mg/mL of desoximetasone in methanol Sample solution: 0.04 mg/mL of desoximetasone from Sample stock solution, and a (1:1) mixture of methanol and acetonitrile [10:90] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm stainless steel column; packing L7 Flow rate: 1 mL/min Injection size: 10 L (injection loop) System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 1.5 Relative standard deviation: NMT 2.0% for five replicate injections Analysis Samples: Sample solution and Standard solution Calculate the percentage of C22H29FO4 in the portion of Desoximetasone taken: Result = (HU/HS) (CS/CU) F 100 = peak height of desoximetasone from the Sample solution = peak height of desoximetasone from the HS Standard solution CS = concentration of USP Desoximetasone RS in the Standard solution (mg/mL) CU = concentration of Desoximetasone in the Sample solution (mg/mL) Acceptance criteria: 97.0%103.0% HU IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% HEAVY METALS, Method II 231: NMT 20 ppm SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 206218; but the range between beginning and end of melting does not exceed 4 OPTICAL ROTATION, Specific Rotation 781S: +107 to +112 Sample solution: 5 mg/mL, in chloroform LOSS ON DRYING 731: Dry it at 105 to constant weight: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Desoximetasone RS
Analysis: Determine the amounts of desogestrel and ethinyl estradiol dissolved as described in Test 1. Tolerances: NLT 80% (Q) of the labeled amount of C22H30O and C20H24O2 is dissolved. [NOTEIf the product complies with this test, the labeling indicates that it meets USP Dissolution Test 2.] UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements for Content Uniformity for both Desogestrel and Ethinyl Estradiol ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers LABELING: When more than one Dissolution test is given, the labeling states the Dissolution test used only if Test 1 is not used. USP REFERENCE STANDARDS 11 USP Desogestrel RS USP Ethinyl Estradiol RS
Desoximetasone
(Comment on this Monograph)id=m23060=Desoximetasone=DMonos.pdf)
376.46 C22H29FO4 Pregna-1,4-diene-3,20-dione, 9-fluoro-11,21-dihydroxy-16methyl-, (11,16)-; 9-Fluoro-11,21-dihydroxy-16-methylpregna-1,4-diene-3,20dione [382-67-2]. DEFINITION Desoximetasone contains NLT 97.0% and NMT 103.0% of C22H29FO4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. THIN-LAYER CHROMATOGRAPHY Standard solution: 10 mg/mL of USP Desoximetasone RS, in chloroform and alcohol (3:1) Sample solution: 10 mg/mL of desoximetasone, in chloroform and alcohol (3:1) Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 20 L Developing solvent system: Chloroform and ethyl acetate (1:1) Spray reagent: 200 mg/mL of p-toluenesulfonic acid in alcohol Analysis Samples: Standard solution and Sample solution Allow the solvent front to move 10 cm beyond the application point. After drying, examine the plate under UV light at 254 nm. Spray the dried plate with Spray reagent. Acceptance criteria: The major spot from the Sample solution corresponds in RF value (0.25) and appearance to that obtained from the Standard solution.
USP 32
Official Monographs / Desoximetasone 37

Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for ethylparaben and desoximetasone are about 1.0 and 2.0, respectively.] Suitability requirements Resolution: NLT 2.0 between the analyte and internal standard peaks Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO4 in the portion of Cream taken: Result = (RU/RS) (CS/CU) 100 = peak response ratios from the Sample solution = peak response ratios from the Sample solution = concentration of USP Desoximetasone RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% RU RS CS PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS PH 791: 4.08.0, in a solution prepared in the following manner. Add 15 mL of boiling water to 3.5 g of the Cream in a 50-mL centrifuge tube, cap the tube, shake vigorously until the cream is uniformly dispersed, then place the tube in a steam bath until the water and oil layers separate completely. Cool, separate the layers, and determine the pH of the aqueous phase. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Desoximetasone RS
Desoximetasone Cream
(Comment on this Monograph)id=m23070=Desoximetasone Cream=D-Monos.pdf) DEFINITION Desoximetasone Cream is Desoximetasone in an emollient cream base. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H29FO4. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 500 g/mL of USP Desoximetasone RS in acetonitrile Sample solution: Evaporate 25 mL of the Sample solution, prepared as directed in the Assay, on a steam bath just to dryness, and dissolve the residue in 2 mL of acetonitrile. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform and ethyl acetate (1:1) Spray reagent: 200 mg/mL of p-toluenesulfonic acid in alcohol Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram in a saturated chamber containing the Developing solvent system. Allow the solvent front to move 10 cm beyond the application point. After drying, examine the plate under UV light at 254 nm. Spray the dried plate with Spray reagent. Acceptance criteria: The major spot from the Sample solution corresponds in RF value (0.25) and appearance to that from the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (65:1:35) Internal standard solution: 0.04 mg/mL of ethylparaben in methanol Standard stock solution: 0.4 mg/mL of USP Desoximetasone RS in methanol Standard solution: Pipet 5 mL of Standard stock solution into a 50-mL centrifuge tube. Add 10.0 mL of Internal standard solution, dilute with methanol to 40.0 mL to obtain a known volume. Sample solution: Transfer an amount of Cream, nominally equivalent to 2 mg of desoximetasone, to a 50-mL centrifuge tube, and add a few 3-mm glass beads. Add 10.0 mL of Internal standard solution, and 30 mL of methanol. Tightly cap the centrifuge tube, and immerse it for 10 min in a bath maintained at 65. Remove the tube from the bath, and immediately vortex at high speed for 30 s. Return the tube to the hot water bath for 5 min, remove it from the bath, and immediately vortex for 30 s. Repeat the procedure, then cool the tube in an ice-bath held at 10 until no further flocculent precipitation occurs. Centrifuge, and use the supernatant. Chromatographic system (See Chromatography 621, System Suitability.)
Desoximetasone Gel
(Comment on this Monograph)id=m23075=Desoximetasone Gel=D-Monos.pdf) DEFINITION Desoximetasone Gel contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H29FO4. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 1 mg/mL of USP Desoximetasone RS in methanol Sample solution: Nominally equivalent to 100 g of desoximetasone from Gel, in a 15-mL centrifuge tube. Add 3 mL of acetonitrile, sonicate for approximately 1 min, centrifuge, and transfer the clear supernatant to another 15-
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Desoximetasone / Official Monographs

Acceptance criteria: 18.0%24.0% (w/w)
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mL centrifuge tube. Evaporate the solution under nitrogen at a temperature between 35 and 45 to dryness. Dissolve the residue in 100 L of methanol, using a sonicator. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture and an area of preadsorbent material Developing solvent system: Acetone and chloroform (1:1) Analysis Samples: Standard solution and Sample solution Streak separately the entire Sample solution and 100 L of the Standard solution on a thin-layer chromatographic plate. Allow the streaks to dry, and develop the chromatogram. Allow the solvent front to move NLT 10 cm beyond the origin. After drying, examine the plate under UV light at 254 nm. Acceptance criteria: The RF value of the principal spot obtained in the chromatogram of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (65:1:35). Adjust the ratio, if necessary, so that the retention time of desoximetasone is about 8 min. Standard stock solution: 0.5 mg/mL of USP Desoximetasone RS in methanol Standard solution: 0.025 mg/mL of USP Desoximetasone RS from Standard stock solution in methanolic calcium chloride dihydrate solution (15 mg/mL) Sample solution: Equivalent to 0.025 mg/mL of desoximetasone from Gel, in methanolic calcium chloride dihydrate solution (15 mg/mL) [NOTESonicate to disperse the gel. Use the clear supernatant.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution (five replicate injections) Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO4 in the portion of Gel taken: Result = (rU/rS) (CS/CU) 100 = peak response of desoximetasone obtained from the Sample solution = peak response of desoximetasone obtained from rS the Standard solution CS = concentration of USP Desoximetasone RS in the Standard solution (mg/mL) CU = nominal concentration of dexosimetasone in the Sample solution (unit/mL) Acceptance criteria: 90.0%110.0% rU OTHER COMPONENTS ALCOHOL CONTENT (See Alcohol Determination 611) Sample solution: 50 mg/mL of Gel in methanol Analysis: Determine the alcohol content of the specimen thus prepared by the Method IIGasLiquid Chromatographic Method, using isopropyl alcohol as the internal standard and using methanol in place of water as the solvent.
PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes, at controlled room temperature. USP REFERENCE STANDARDS 11 USP Desoximetasone RS
Desoximetasone Ointment
(Comment on this Monograph)id=m23079=Desoximetasone Ointment=D-Monos.pdf) DEFINITION Desoximetasone Ointment contains NLT 90.0% and NMT 110.0% of the labeled amount of desoximetasone (C22H29FO4). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 0.5 mg/mL of USP Desoximetasone RS in acetonitrile Sample solution: Equivalent to 5 mg of desoximetasone from Ointment to a 50-mL centrifuge tube. Add 20 mL of hexane, heat gently to 60, and shake until the Ointment is completely dispersed. Add 8 mL of acetonitrile, insert the stopper in the tube, and shake vigorously for 5 min. Cool to room temperature, and centrifuge until the lower layer is clear. Transfer the lower layer to a 10-mL volumetric flask, and dilute with acetonitrile to volume. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Ethyl acetate and chloroform (4:1) Spray reagent: 200 mg/mL of p-toluenesulfonic acid in alcohol Analysis Samples: Sample solution and Standard solution Allow the spots to dry, and develop the plate in a saturated chamber containing the Developing solvent system, until the solvent front has moved three-fourths of the length of the plate. Remove the plate, and allow to air-dry. Examine under short-wavelength UV light. Spray the dried plate with Spray reagent. Heat the plate at 100 for 5 min, and examine under long-wavelength UV light. Acceptance criteria: The RF value and appearance (brownish-yellow fluorescent spot) of the principal spot from the Sample solution correspond to those of the principal spot from the Standard solution. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (65:1:35) Standard stock solution: 0.4 mg/mL of USP Desoximetasone RS in methanol Standard solution: 0.04 mg/ml of USP Desoximetasone RS from Standard stock solution and a 1:1 mixture of methanol and spectrophotometric acetonitrile that is saturated with nheptane (1:9) Sample solution: Nominally equivalent to 2 mg of desoximetasone from Ointment to a 50-mL centrifuge tube. Add 20 mL of n-heptane that has been previously saturated with spectrophotometric acetonitrile, and heat gently with occasional shaking until the Ointment is completely
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dispersed. Allow to cool slightly, and extract with a 10-mL portion of spectrophotometric acetonitrile. Shake vigorously, centrifuge, remove the bottom layer of acetonitrile with a syringe and needle, and transfer to a 50-mL volumetric flask. Using the same needle and syringe, extract the desoximetasone with successive 10-mL and 8-mL portions of acetonitrile, combining all acetonitrile layers in the 50-mL flask. Dilute with methanol nearly to volume, and allow the solution to reach room temperature. Dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 15-cm; packing L7 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 5.0 between the analyte and solvent peaks Tailing factor: NMT 2.0 for the analyte peak Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO4 in the portion of Ointment taken: Result = (rU/rS) (CS/CU) 100 = peak response of the Sample solution = peak response of the Standard solution = concentration of USP Desoximetasone RS in the Standard solution (mg/mL) = nominal concentration of desoximetasone in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes at a controlled room temperature. USP REFERENCE STANDARDS 11 USP Desoximetasone RS rU rS CS
Official Monographs / Desoxycorticosterone 39

IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: At 240 nm Sample solution: 10 g/mL in alcohol Acceptance criteria: Absorptivities calculated on the dried basis, do not differ by more than 3.0% ASSAY PROCEDURE Standard solution: Prepare as directed under Assay for Steroids 351, Standard Preparation, using USP Desoxycorticosterone Acetate RS. Sample stock solution: 0.5 mg/mL of Desoxycorticosterone Acetate in alcohol Sample solution: Pipet 5 mL of Sample stock solution into a 250-mL volumetric flask, and add alcohol to volume. Pipet 20 mL of this solution into a glass-stoppered, 50-mL conical flask. Analysis: Proceed as directed under Assay for Steroids 351, Analysis. Calculate the percentage of C23H32O4 in the Desoxycorticosterone Acetate taken: Result = (AU/AS) (CS/CU) 100 AU = absorbance of the Sample solution AS = absorbance of the Standard solution = concentration of the Standard solution (mg/mL) CS = concentration for the Sample solution (mg/mL) CU Acceptance criteria: 97.0%103.0% SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 155161 OPTICAL ROTATION, Specific Rotation 781S: +171 to +179 Sample solution: 10 mg/mL, undried, in dioxane LOSS ON DRYING 731: Dry it in vacuum over silica gel for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. Store at 25; excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Desoxycorticosterone Acetate RS
Desoxycorticosterone Acetate Injection

(Comment on this Monograph)id=m23130=Desoxycorticosterone Acetate Injection=D-Monos.pdf) DEFINITION Desoxycorticosterone Acetate Injection is a sterile solution of Desoxycorticosterone Acetate in vegetable oil. It contains NLT 90.0% and NMT 115.0% of the labeled amount of C23H32O4. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Evaporate 25 mL of the Sample solution from the Assay on a steam bath just to dryness, and dissolve the residue in 1 mL of chloroform. ASSAY PROCEDURE Alcohol-isooctane and Isooctane-alcohol: Shake equal volumes of 90% alcohol and isooctane in a separator for 10 to 15 min, and allow to separate. Withdraw the layers into
Desoxycorticosterone Acetate
(Comment on this Monograph)id=m23120=Desoxycorticosterone Acetate=DMonos.pdf)
C23H32O4 Pregn-4-ene-3,20-dione, 21-(acetyloxy)-; 11-Deoxycorticosterone acetate [56-47-3].
372.50
DEFINITION Desoxycorticosterone Acetate contains NLT 97.0% and NMT 103.0% of C23H32O4, calculated on the dried basis.
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Desoxycorticosterone / Official Monographs
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alcohol to obtain a solution having a concentration of 10 g/mL. Pipet 20 mL of this solution into a glass-stoppered, 50-mL conical flask. Sample solution: 100 mg of the powder from NLT 10 Pellets, dissolved in sufficient alcohol to make 200.0 mL Transfer 5.0 mL of this solution to a 250-mL volumetric flask, dilute with alcohol to volume. Transfer 20.0 mL of this solution to a glass-stoppered, 50-mL conical flask. Analysis: Proceed as directed in Assay for Steroids 351, Procedure. Calculate the percentage of C23H32O4 in the portion of Pellets taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Desoxycorticosterone Acetate RS in the Standard solution (g/mL) = nominal concentration of desoxycorticosterone in CU the Sample solution (g/mL) Acceptance criteria: 97.0%103.0% AU AS CS PERFORMANCE TESTS WEIGHT VARIATION: Weigh 5 Pellets singly, and calculate the average weight. The average weight is NLT 95% and NMT 105% of the labeled weight of C23H32O4, and each Pellet weighs NLT 90% and NMT 110% of the labeled weight of C23H32O4. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 155161 OPTICAL ROTATION, Specific Rotation 781S: +171 to +179 Sample solution: 10 mg/mL, undried, in dioxane STERILITY TESTS 71: Meet the requirements SOLUBILITY IN ALCOHOL: A solution of 25 mg of powdered Pellets in 1 mL of alcohol is clear and practically free from insoluble residue. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers suitable for maintaining sterile contents, holding one pellet each. USP REFERENCE STANDARDS 11 USP Desoxycorticosterone Acetate RS
separate containers, designating the lower layer as alcoholisooctane and the upper layer as isooctane-alcohol. Standard solution: Dissolve in alcohol a suitable quantity of USP Desoxycorticosterone Acetate RS, and dilute with alcohol to obtain a solution having a concentration of 10 g/mL. Pipet 20 mL of this solution into a glass-stoppered, 50-mL conical flask. Sample solution: Equivalent to 5 mg of desoxycorticosterone acetate from Injection, in a separator containing 50 mL of Isooctane-alcohol. Extract with six 20mL portions of Alcohol-isooctane, receiving the extracts in a 250-mL volumetric flask, dilute with Alcohol-isooctane to volume. Pipet 10 mL of this solution into a glass-stoppered, 50-mL conical flask, evaporate on a steam bath with the aid of a gentle current of air just to dryness, and dissolve the residue in 20.0 mL of alcohol. Analysis: Proceed as directed in Assay for Steroids 351, Procedure. Calculate the percentage of C23H32O4 in each mL of the Injection taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Desoxycorticosterone Acetate RS in the Standard solution (g/mL) CU = nominal concentration of desoxycorticosterone acetate in the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS OTHER REQUIREMENTS: It meets the requirements under Injections 1. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 71.4 USP Endotoxin Unit/mg of desoxycorticosterone acetate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I or Type III glass, protected from light. USP REFERENCE STANDARDS 11 USP Desoxycorticosterone Acetate RS USP Endotoxin RS AU AS CS
Desoxycorticosterone Acetate Pellets

(Comment on this Monograph)id=m23140=Desoxycorticosterone Acetate Pellets=D-Monos.pdf) DEFINITION Desoxycorticosterone Acetate Pellets are sterile pellets composed of Desoxycorticosterone Acetate in compressed form, without the presence of any binder, diluent, or excipient. They contain NLT 97.0% and NMT 103.0% of C23H32O4. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: At 240 nm Sample solution: 10 g/mL in alcohol Acceptance criteria: Absorptivities do not differ by more than 3.0%, calculated on the dried basis ASSAY PROCEDURE Standard solution: Dissolve in alcohol a suitable quantity of USP Desoxycorticosterone Acetate RS, and dilute with
Desoxycorticosterone Pivalate
(Comment on this Monograph)id=m23170=Desoxycorticosterone Pivalate=DMonos.pdf) 414.58 C26H38O4 Pregn-4-ene-3,20-dione, 21-(2,2-dimethyl-1-oxopropoxy)-; 11-Deoxycorticosterone pivalate [808-48-0]. DEFINITION Desoxycorticosterone Pivalate contains NLT 97.0% and NMT 103.0% of C26H38O4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: At 241 nm Sample solution: 20 g/mL in methanol Acceptance criteria: Absorptivities do not differ by more than 3.0%, calculated on the dried basis
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ASSAY PROCEDURE Mobile phase: Methanol and water (4:1) Internal standard solution: 2 mg/mL of desoxycorticosterone in methanol Standard solution: Transfer 12.5 mg of USP Desoxycorticosterone Pivalate RS to a 25-mL volumetric flask, and add 20 mL of methanol. Add 2.5 mL of Internal standard solution, dilute with methanol to volume, and mix to obtain a concentration of 0.5 mg/mL of USP Desoxycorticosterone Pivalate RS. Sample solution: Transfer 50 mg of desoxycorticosterone pivalate to a 100-mL volumetric flask, and add 80 mL of methanol. Add 10.0 mL of Internal standard solution, and dilute with methanol to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1.5 mL/min Injection size: 25 L System suitability Sample: Standard solution Suitability requirements Resolution: NLT 2.0 between the analyte and internal standard peaks Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution [NOTEThe relative retention times for desoxycorticosterone acetate and desoxycorticosterone pivalate are about 0.5 and 1.0, respectively.] Calculate the percentage of C26H38O4 in the portion of Desoxycorticosterone Pivalate taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Desoxycorticosterone Pivalate RS in the Standard solution (mg/mL) = concentration of Desoxicorticosterone Pivalate in CU the Sample solution (mg/mL) Acceptance criteria: 97.0%103.0% RU RS CS SPECIFIC TESTS MELTING RANGE OR TEMPERATURE 741: 200206 OPTICAL ROTATION, Specific Rotation 781S: +155 to +163 Sample solution: 10 mg/mL, in dioxane LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. Store at 25; excursions permitted between 15 and 30. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Desoxycorticosterone Pivalate RS
Official Monographs / Desoxycorticosterone 41
Desoxycorticosterone Pivalate Injectable Suspension

(Comment on this Monograph)id=m23175=Desoxycorticosterone Pivalate Injectable Suspension=D-Monos.pdf) DEFINITION Desoxycorticosterone Pivalate Injectable Suspension is a sterile suspension of Desoxycorticosterone Pivalate in an aqueous medium. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C26H38O4. IDENTIFICATION PROCEDURE Sample: Centrifuge a portion of the Injectable Suspension, decant the supernatant, wash the residue by stirring with several successive portions of water, centrifuging and decanting each time, and finally dry the residue at 105. The residue meets the following requirements. Analysis 1: Melting point Acceptance criteria 1: Melts 198204 Analysis 2: Dissolve 5 mg in 2 mL of sulfuric acid. Acceptance criteria 2: The solution is yellowish with greenish fluorescence. Analysis 3: Dilute the solution obtained from Analysis 2 with 2 mL of water. Acceptance criteria 3: The color changes to a dark redblue and, on further dilution with 2 mL of water, the color is discharged. ASSAY PROCEDURE Mobile phase: Methanol and water (4:1) Internal standard solution: 2 mg/mL of desoxycorticosterone in methanol Standard solution: Transfer 12.5 mg of USP Desoxycorticosterone Pivalate RS to a 25-mL volumetric flask, and add 20 mL of methanol. Add 2.5 mL of the Internal standard solution, dilute with methanol to volume and mix to obtain a concentration of 0.5 mg/mL of USP Desoxycorticosterone Pivalate RS. Sample solution: Transfer a nominal equivalent of 125 mg of desoxycorticosterone pivalate from Injectable Suspension to a 250-mL volumetric flask. Add 200 mL of methanol, and sonicate to dissolve. Add 25.0 mL of Internal standard solution, dilute with methanol to volume. Centrifuge a 20mL portion at high speed for 5 min. Filter the supernatant through a 5-m disk, discarding the first 5 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1.5 mL/min Injection size: 25 L System suitability Sample: Standard solution [NOTEThe relative retention times for desoxycorticosterone acetate and for desoxycorticosterone pivalate are about 0.5 and 1.0, respectively.]
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Standard stock solution: 7.5 mg/mL of USP Dexamethasone RS in methanol Standard solution: 0.3 mg/mL from Standard stock solution in Mobile phase Sample stock solution: Using 30 mg, prepare a 7.5 mg/mL solution of Dexamethasone in methanol. Sample solution: 0.3 mg/mL from Sample stock solution in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 25-cm; packing L7 Injection size: 1530 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 3.0% (5 replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in the portion of Dexamethasone taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) = concentration of dexamethasone in the Sample CU solution (mg/mL) Acceptance criteria: 97.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% from 250 mg Organic Impurities PROCEDURE Solution A: 1.32 mg/mL in water, adjust with formic acid to a pH of 3.6 Mobile phase: Acetonitrile and Solution A (33:67) Sample stock solution: 1.8 mg/mL of Dexamethasone in acetonitrile Sample solution: 0.6 mg/mL from Sample stock solution and Solution A (33:67) Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L11 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Sample solution Suitability requirements Column efficiency: NLT 5000 theoretical plates Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Dexamethasone taken: Result = (rU/rT) 100 rU rT = peak response for each impurity = sum of the responses of all peaks rU rS CS
Suitability requirements Resolution: NLT 2.0 between the analyte and internal standard peaks Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C26H38O4 in the portion of Suspension taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio from the Sample solution = peak response ratio from the Standard solution = concentration of USP Desoxycorticosterone Pivalate RS in the Standard solution (mg/mL) = nominal concentration of desoxicorticosterone CU pivalate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU RS CS SPECIFIC TESTS PH 791: 5.07.0 OTHER REQUIREMENTS: It meets the requirements under Injections 1. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 2.78 USP Endotoxin Unit/mg of desoxycorticosterone pivalate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, protected from light. LABELING: Label Suspension to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Desoxycorticosterone Pivalate RS USP Endotoxin RS
Dexamethasone
(Comment on this Monograph)id=m23280=Dexamethasone=DMonos.pdf)
392.47 C22H29FO5 Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17,21-trihydroxy-16methyl-, (11,16)-; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione [50-02-2]. DEFINITION Dexamethasone contains NLT 97.0% and NMT 102.0% of C22H29FO5, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 239 nm Solution: 10 g/mL in methanol Acceptance criteria: Absorptivities calculated on the dried basis do not differ by more than 3.0%. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (3:7) such that at an approximate flow rate of 2 mL/min, the retention time of Dexamethasone is about 7 min
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Acceptance criteria Individual impurities: NMT 1.0% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: + 72 to +80 Sample solution: 10 mg/mL, in dioxane LOSS ON DRYING 731: Dry it at 105 for 3 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dexamethasone RS
Official Monographs / Dexamethasone 43

Analysis: Proceed as directed in Assay for Steroids 351, Procedure, except to allow to stand in the dark for 45 min. Calculate the percentage of C22H29FO5 in each container taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) CU = nominal concentration of dexamethasone in the Sample solution (mg/mL) Acceptance criteria: 90.0%120.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: It meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. OTHER REQUIREMENTS: It meets the requirements for Pressure Test, Minimum Fill, and Leakage Test under Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers 601. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in pressurized containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Dexamethasone RS AU AS CS
Dexamethasone Topical Aerosol

(Comment on this Monograph)id=m23290=Dexamethasone Topical Aerosol=D-Monos.pdf) DEFINITION Dexamethasone Topical Aerosol is Dexamethasone in a suitable lotion base mixed with suitable propellants in a pressurized container. Dexamethasone Topical Aerosol delivers NLT 90.0% and NMT 120.0% of the labeled amount of C22H29FO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 500 g/mL of USP Dexamethasone RS in chloroform Sample solution: Evaporate 5 mL of the Sample solution, obtained as directed in the Assay, on a steam bath just to dryness, and dissolve the residue in 1 mL of chloroform. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel Application volume Sample solution: 100 L Standard solution: 10 L Developing solvent system: Chloroform and diethylamine (2:1) Spray reagent: dilute sulfuric acid (1 in 2) Analysis Samples: Standard solution and Sample solution Locate the spots on the plate by lightly spraying with Spray reagent and heating on a hot plate or under a heat lamp until spots appear. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Standard solution: Dissolve in alcohol a suitable quantity of USP Desoxycorticosterone Acetate RS, and dilute with alcohol to obtain a solution having a concentration of 10 g/mL. Pipet 20 mL of this solution into a glass-stoppered, 50-mL conical flask. Sample solution: Shake the Topical Aerosol container gently, and invert, immersing the valve in 75 mL of alcohol contained in a 400-mL beaker. Actuate the valve by pushing against the bottom of the beaker. Remove the container at 15-s intervals, shake gently, and allow the container to warm to room temperature. Continue spraying until the contents of the container are exhausted. Transfer the alcohol solution to a 100-mL volumetric flask, dilute with alcohol to volume. Dilute a volume of this solution, nominally equivalent to 1 mg of dexamethasone, with alcohol to 100.0 mL. Transfer 20.0 mL of this solution to a glassstoppered, 50-mL flask.
Dexamethasone Elixir
(Comment on this Monograph)id=m23310=Dexamethasone Elixir=D-Monos.pdf) DEFINITION Dexamethasone Elixir contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H29FO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Solution A: Methylene chloride and methanol (1:1) Standard solution: 500 g/mL of USP Dexamethasone RS in Solution A Sample solution: Evaporate 9 mL of the Sample solution, prepared as directed in the Assay, on a steam bath just to dryness, and dissolve the residue in 2 mL of Solution A. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Chloroform, acetone, and glacial acetic acid (80:40:1) Analysis Samples: Standard solution and Sample solution Proceed as directed under General Chapter. Locate the spots by viewing under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot from the Sample solution corresponds to that from the Standard solution. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (1:2) Standard solution: 0.1 mg/mL of USP Dexamethasone RS in dilute methanol (1 in 2) Sample solution: Nominally equivalent to 0.1 mg/mL of dexamethasone from Elixir (freshly mixed and free from air
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measuring 30 1.5-cm, equipped with a polytef stopcock. Fill the tube half-full with Mobile solvent. Mix 8 g of chromatographic siliceous earth with 8 mL of methanol and water (1:1). Transfer successive portions of the mixture to the column, emptying and adding Mobile solvent after each addition to pack the column. Drain the column to a layer of Mobile solvent 1 cm above the absorbant. Pack a second tube to provide a blank, and proceed as directed for Sample solution, but omit the specimen. Standard solution: 10 g/mL of USP Dexamethasone RS in alcohol Sample solution: Equivalent to 0.5 mg of dexamethasone from Gel, in a 100-mL beaker. Add 1 g of chromatographic siliceous earth. Transfer the mixture to the column, wash the beaker with small portions of Mobile solvent, adding them to the column. Adjust the flow rate to 2 mL per minute, discarding the eluate. Elute with four additional 25-mL portions of Mobile solvent, and discard. Rinse the sample beaker with two 25-mL portions of methylene chloride and transfer to the column, collecting the eluate in a suitable beaker. Elute the column with six additional 25-mL portions of methylene chloride, combining the eluates and evaporating with a gentle current of air to dryness, dissolve the residue in alcohol, and transfer to a 50-mL volumetric flask. Wash the beaker with successive 5-mL portions of alcohol, collecting the washings in the flask, and dilute with alcohol to volume. Centrifuge or filter, and then pipet 10 mL of this solution, 10 mL of the Standard solution, 10 mL of the column blank solution, and 10 mL of alcohol to provide a reagent blank to separate flasks. Analysis: Proceed as directed under Assay for Steroids 351, except to allow to stand in the dark for 45 min. Calculate the percentage of C22H29FO5 in the portion of Gel taken: Result = (AU ACB/AS ARB) (Cs/Cu) 100 absorbance of the Sample solution absorbance of the column blank preparation absorbance of the Standard solution absorbance of the reagent blank solution concentration of USP Dexamethasone RS in the Standard solution (g/mL) = nominal concentration of dexamethasone in the Cu Sample solution (g/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes. Keep tightly closed. Avoid exposure to temperatures exceeding 30. USP REFERENCE STANDARDS 11 USP Dexamethasone RS AU ACB AS ARB Cs = = = = =
bubbles) diluted with water, and filter through a suitable membrane filter Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 525 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 3.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in each mL of the Elixir taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) = nominal concentration of dexamethasone in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS OTHER COMPONENTS ALCOHOL DETERMINATION, Method II 611: 3.8%5.7% of C2H5OH, n-propyl alcohol being used as the internal standard ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dexamethasone RS
Dexamethasone Gel
(Comment on this Monograph)id=m23320=Dexamethasone Gel=D-Monos.pdf) DEFINITION Dexamethasone Gel contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H29FO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Standard solution: 500 g/mL of USP Dexamethasone RS in chloroform Sample solution: Evaporate 25 mL of the Sample solution, prepared as directed in the Assay, on a steam bath just to dryness, and dissolve the residue in 0.5 mL of chloroform. Use the chloroform extract. Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 10 L Developing solvent system: Chloroform and diethylamine (2:1) Analysis: Proceed as directed in the chapter. Locate the spots on the plate by lightly spraying with dilute sulfuric acid (1 in 2) and heating. Acceptance criteria: The RF value of the principal spot obtained from the Sample solution corresponds to that obtained from the Standard solution. ASSAY PROCEDURE Mobile solvent: Isooctane and methylene chloride (9:1) Chromatographic columns: Tamp a pledget of glass wool at the constriction of a glass chromatographic tube
Dexamethasone Injection
(Comment on this Monograph)id=m23325=Dexamethasone Injection=D-Monos.pdf) DEFINITION Dexamethasone Injection is a sterile solution of Dexamethasone in Water for Injection. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H29FO5. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Sample solution: Quantity of Injection equivalent to 5 mg of dexamethasone to a 50-mL separator
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Add 10 mL of water, and extract with two 20-mL portions of chloroform. Filter the lower layers through chloroformsaturated cotton into a 50-mL conical flask, and evaporate to dryness. Dissolve the residue in 10 mL of chloroform. Developing solvent system: Methylene chloride and methanol (180:16) Analysis: Visualize the spots using a 1 in 5 solution of ptoluenesulfonic acid in a mixture of alcohol and propylene glycol (9:1), followed by heat. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (30:70) System suitability solution: 0.3 mg/mL of USP Dexamethasone RS, 1.35 mg/mL of benzyl alcohol, 0.27 mg/mL of methylparaben, and 0.03 mg/mL of propylparaben in Mobile phase Standard solution: Quantitatively dissolve a weighed amount of USP Dexamethasone RS in methanol to obtain a stock solution having a known concentration of 7.5 mg/mL. Transfer 4.0 mL to a 100-mL volumetric flask, and dilute with Mobile phase to volume to obtain a solution having a known concentration of 0.3 mg of USP Dexamethasone RS/mL. Sample solution: Injection equivalent to 0.3 mg/mL of dexamethasone in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; 5-m packing L7 Flow rate: 2 mL/min Injection size: 20 L System suitability Samples: System suitability solution and Standard solution [NOTEThe relative retention times for benzyl alcohol, methylparaben, dexamethasone, and propylparaben are 0.4, 0.5, 1.0, and 1.4, respectively, System suitability solution.] Suitability requirements Resolution: NLT 3 between the neighboring peaks for benzyl alcohol and methylparaben, methylparaben and dexamethasone, and dexamethasone and propylparaben in the System suitability solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution CU (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 4.05.5 INJECTIONS 1: It meets the requirements. STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. PARTICULATE MATTER IN INJECTIONS 788: It meets the requirements for small-volume injections.

BACTERIAL ENDOTOXINS TEST 85: Contains NMT 21.0 USP Endotoxin Unit/mg of dexamethasone ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in light-resistant, singledose or multiple-dose containers, preferably of Type I glass. LABELING: Label it to indicate that it is for veterinary use only. USP REFERENCE STANDARDS 11 USP Dexamethasone RS USP Endotoxin RS
Dexamethasone Ophthalmic Suspension

(Comment on this Monograph)id=m23330=Dexamethasone Ophthalmic Suspension=D-Monos.pdf) DEFINITION Dexamethasone Ophthalmic Suspension is a sterile, aqueous suspension of dexamethasone containing a suitable antimicrobial preservative. It may contain suitable buffers, stabilizers, and suspending and viscosity agents. It contains NLT 90.0% and NMT 110.0% of the labeled amount of C22H29FO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture. Standard solution: 500 g/mL of USP Dexamethasone RS in chloroform Sample solution: Transfer a volume of Ophthalmic Suspension, equivalent to about 2.5 mg of dexamethasone, to a test tube, add 5 mL of chloroform, shake, and centrifuge. Application volume: 10 L Developing solvent system: Solvent A as directed under Single-Steroid Assay 511 Analysis: Develop the chromatogram, mark the solvent front, and locate the spots on the plate by spraying with a solution (1 in 5) of p-toluenesulfonic acid in a mixture of 9 volumes of alcohol and 1 volume of propylene glycol, and heating until spots appear. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (40:60) Standard solution: 0.12 mg/mL of USP Dexamethasone RS in Mobile phase Sample solution: Equivalent to 0.12 mg/mL of dexamethasone from Ophthalmic Suspension (freshly mixed and free from air bubbles), in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L1 Flow rate: 2 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 1750 theoretical plates from the analyte peak, Standard solution Tailing factor: NMT 3.0 for the analyte peak, Standard solution Relative standard deviation: NMT 3.0% of dexamethasone, Standard solution
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Sample solution: 0.04 mg/mL of dexamethasone in Diluent Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 2000 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in the portion of Oral Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) CU = nominal concentration of dexamethasone in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS OTHER COMPONENTS ALCOHOL DETERMINATION, Method II 611 (if present): 27.0%33.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For oral solution packaged in single-unit containers: meets the requirements DELIVERABLE VOLUME 698: For oral solution packaged in multiple-unit containers: meets the requirements SPECIFIC TESTS PH 791: 2.74.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. LABELING: Label concentrated Oral Solution to state that the term Concentrate is to appear apart from and immediately after the official title in prominent boldface type. Label concentrated Oral Solution also to indicate that it is to be diluted to appropriate strength with a suitable diluent prior to administration unless produced for dispensing with instructions for administration by a calibrated dropper or syringe. USP REFERENCE STANDARDS 11 USP Dexamethasone RS
Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in each mL of the Ophthalmic Suspension taken: Result = (rU/rS) (CS/CU) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) = concentration of dexamethasone in the Sample CU solution (mg/mL) Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 5.06.0 STERILITY TESTS 71:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dexamethasone RS
Dexamethasone Oral Solution

(Comment on this Monograph)id=m23335=Dexamethasone Oral Solution=D-Monos.pdf) DEFINITION Dexamethasone Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of dexamethasone (C22H29FO5). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY IDENTIFICATION TEST 201 Sample solution: Oral Solution, equivalent to 5 mg of dexamethasone, in a 50-mL separator, add 10 mL of water, and extract with two 20-mL portions of chloroform. Filter the lower layers through chloroform-saturated cotton into a 50-mL conical flask, and evaporate to dryness. Dissolve the residue in 10 mL of chloroform. Developing solvent system: Methylene chloride and methanol (180:16) Analysis: Visualize the spots, using a solution (1 in 5) of ptoluenesulfonic acid in a mixture of alcohol and propylene glycol (9:1) followed by heat. ASSAY PROCEDURE Mobile phase: Methanol, glacial acetic acid, and water (55:2:43) Diluent: Methanol and water (1:1) Standard solution: 0.04 mg/mL of USP Dexamethasone RS in Diluent
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Dexamethasone Tablets
(Comment on this Monograph)id=m23340=Dexamethasone Tablets=D-Monos.pdf) DEFINITION Dexamethasone Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C22H29FO5. IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Standard solution: 500 g/mL of Dexamethasone RS in chloroform Sample solution: Evaporate 10 mL of the methanol extract of Tablets obtained as directed under Sample solution in the Assay on a steam bath just to dryness, and dissolve the residue in 1 mL of chloroform. Mode: TLC Application volume Sample solution: 10 L Standard solution: 20 L Analysis Samples: Standard solution and Sample solution Develop the chromatogram in Solvent A as directed under Single-Steroid Assay 511. Mark the solvent front, and locate the spots on the plate by visualizing under shortwavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile solvent: Acetonitrile and water (1:3) Standard solution: 0.1 mg/mL of USP Dexamethasone RS in dilute methanol (1 in 2) Sample solution: Transfer an equivalent to 5 mg of dexamethasone, from finely powdered Tablets (NLT 10), to a 50-mL volumetric flask, and add 30 mL of dilute methanol (1 in 2). Sonicate the flask for 2 min, shake by mechanical means for 30 min, and dilute with the same solvent to volume. Pass a portion of the mixture through a suitable filter to obtain a clear filtrate. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 30-cm; packing L1 Injection size: 525 L System suitability Sample: Standard solution [NOTEThe retention time of dexamethasone is between 3 and 6 min.] Suitability requirement Relative standard deviation: NMT 3.0% (five replicate injections) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) = nominal concentration of the Sample solution (mg/mL)
PERFORMANCE TESTS DISSOLUTION 711 Medium: Dilute hydrochloric acid (1 in 100); 500 mL Apparatus 1: 100 rpm Time: 45 min Detector: UV nm Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Standard solution: Prepare as directed for Standard solution under Assay for Steroids 351, using USP Dexamethasone RS. Analysis: Extract a filtered aliquot of Dissolution Medium, equivalent to 200 g of dexamethasone, with three 15-mL portions of chloroform. Evaporate the combined chloroform extracts on a steam bath just to dryness, cool, and dissolve the residue in 20 mL of alcohol. Proceed as directed for Procedure under Assay for Steroids 351, except to allow to stand in the dark for 45 min. Calculate the percentage of C22H29FO5 dissolved: Result = (AU/AS) CS V (100/L) = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) V = volume of the aliquote used for exraction (mL) L = label claim (mg/tablet) Tolerances: NLT 70% (Q) of the labeled amount of C22H29FO5 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements Procedure for content uniformity Standard solution: Prepare as directed for Standard Solution under Assay for Steroids 351, using USP Dexamethasone RS. Sample solution: Place 1 Tablet in a separator with 15 mL of water, and swirl to disintegrate the Tablet completely. Extract with four 10-mL portions of chloroform, filtering each portion through chloroform-washed cotton into a 50mL volumetric flask, and add chloroform to volume. Pipet a volume of this solution, equivalent to 200 g of dexamethasone into a glass-stoppered, 50-mL conical flask, evaporate the chloroform on a steam bath just to dryness, cool, and dissolve the residue in 20.0 mL of alcohol. Use this where Sample solution is specified in the Analysis. Analysis: Proceed as directed for Procedure under Assay for Steroids 351, except to allow to stand in the dark for 45 min. Calculate the percentage of total steroids, as C22H29FO5, in the Tablet: AU AS CS Result = (AU/AS) CS V (100/L) AU AS CS V L = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (mg/mL) = volume of the Sample solution used for exraction (mL) = label claim (mg/tablet)
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dexamethasone RS
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Dexamethasone Acetate
(Comment on this Monograph)id=m23370=Dexamethasone Acetate=D-Monos.pdf) C24H31FO6.H2O 452.51 Pregna-1,4-diene-3,20-dione, 21-(acetyloxy)-9-fluoro-11,17dihydroxy-16-methyl-, (11,16)-monohydrate; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 21-acetate monohydrate [55812-90-3]. Anhydrous 434.51 [1177-87-3]. DEFINITION Dexamethasone Acetate contains one molecule of water of hydration or is anhydrous. It contains NLT 97.0% and NMT 102.0% of C24H31FO6, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 239 nm Standard solution: 15 g/mL in methanol Absorptivities: Calculated on the dried basis, do not differ by more than 3.0% ASSAY PROCEDURE Mobile phase: Acetonitrile and water (45:55). Filter and degas. Solution A: Transfer 3 mL of 1 N sodium hydroxide, 138 mL of 0.5 N potassium chloride, and 50 mL of 0.5 M monobasic potassium phosphate to a 1-L volumetric flask, and dilute with water to volume. Diluent: Acetonitrile and Solution A (1:1) Standard solution: 25 mg of USP Dexamethasone Acetate RS to a 250-mL volumetric flask. Add 100 mL of Diluent, and sonicate until a clear solution is obtained. Dilute with Diluent to volume. Sample solution: 25 mg of Dexamethasone Acetate to a 250-mL volumetric flask. Add 100 mL of Diluent, and sonicate until a clear solution is obtained. Dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 2.0 Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% for replicate injections Analysis Samples: Standard solution and Sample solution Calculate the percentage of C24H31FO6 in the portion of Dexamethasone Acetate taken: Result = (rU/rS) (CS/CU) 100 rU rS CS CU = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Dexamethasone Acetate RS in the Standard solution (mg/mL) = nominal concentration of dexamethasone acetate in the Sample solution (mg/mL)
IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE Solution A: 1.32 g/L of ammonium formate, adjust with formic acid to a pH of 3.6 Mobile phase: Acetonitrile and Solution A (2:3) Sample solution: Transfer 200 mg of Dexamethasone Acetate to a 100-mL volumetric flask, dissolve in and dilute with acetonitrile to volume, and mix. Transfer 40 mL of this solution to a 100-mL volumetric flask, and dilute with Solution A to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L11 Flow rate: 1 mL/min Injection size: 10 L System suitability Sample: Sample solution Suitability requirements Column efficiency: NLT 5400 theoretical plates, Sample solution Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Dexamethasone Acetate taken: Result = (ri/rs) 100 = peak response for each impurity ri = sum of the responses of all peaks rs Acceptance criteria Individual impurities: NMT 1.0% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +82 to +88 Sample solution: 10 mg/mL, in dioxane LOSS ON DRYING 731: Dry in a vacuum at 105 for 3 h: the hydrous form loses between 3.5% and 4.5%, and the anhydrous form NMT 0.4%, of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at 25, excursions permitted between 15 and 30. LABELING: Label it to indicate whether it is hydrous or anhydrous. USP REFERENCE STANDARDS 11 USP Dexamethasone Acetate RS
Dexamethasone Acetate Injectable Suspension

(Comment on this Monograph)id=m23390=Dexamethasone Acetate Injectable Suspension=D-Monos.pdf) DEFINITION Dexamethasone Acetate Injectable Suspension is a sterile suspension of Dexamethasone Acetate in Water for Injection. It contains an amount of dexamethasone acetate monohydrate (C24H31FO6 H2O) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of dexamethasone (C22H29FO5).
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IDENTIFICATION INFRARED ABSORPTION 197M [NOTEDo not use heat to dry the specimen. Total or partial dehydration may occur.] Standard: Preparation solution of undried USP Dexamethasone Acetate RS as described in Sample, beginning with to a fine-porosity Sample: Transfer the contents of a well-shaken container of Injectable Suspension to a fine-porosity, sintered-glass vacuum filter, filter, and wash with several 10-mL portions of water. Remove the powder from the filter and allow to airdry. ASSAY PROCEDURE Mobile phase: Acetonitrile and water (45:55) Solution A: 0.5 N potassium chloride, 0.5 M monobasic potassium phosphate, 1 N sodium hydroxide, and water (138:50:3:809) Diluent: Acetonitrile and Solution A (1:1) Standard solution: 0.09 mg/mL of USP Dexamethasone Acetate RS in Diluent Sample solution: Add well-shaken Injectable Suspension, equivalent to 40 mg of dexamethasone, to a 100-mL volumetric flask. Add 75 mL of Diluent, and sonicate until a clear solution is obtained. Dilute with Diluent to volume. Transfer 10.0 mL of this solution to a 50-mL volumetric flask, and dilute with Diluent to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; 10-m packing L1 Flow rate: 2 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Capacity factor: NLT 2.0 Column efficiency: NLT 1500 theoretical plates Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in the portion of Injectable Suspension taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dexamethasone Acetate RS in the Standard solution (mg/mL) = concentration of dexamethasone in the Sample CU solution (mg/mL) = molecular weight of dexamethasone, 392.47 Mr1 = molecular weight of anhydrous dexamethasone Mr2 acetate, 434.51 Acceptance criteria: 90.0%110.0% rU rS CS SPECIFIC TESTS PH 791: 5.07.5 OTHER REQUIREMENTS: Meets the requirements under Injections 1 BACTERIAL ENDOTOXINS TEST 85: Contains NMT 21.7 USP Endotoxin Unit/mg of dexamethasone acetate ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass.

USP REFERENCE STANDARDS 11 USP Dexamethasone Acetate RS USP Endotoxin RS
Dexamethasone Sodium Phosphate

(Comment on this Monograph)id=m23450=Dexamethasone Sodium Phosphate=D-Monos.pdf)
C22H28FNa2O8P 516.41 Pregna-1,4-diene-3,20-dione, 9-fluoro-11,17-dihydroxy-16methyl-21-(phosphonooxy)-, disodium salt, (11,16)-; 9-Fluoro-11,17,21-trihydroxy-16-methylpregna-1,4diene-3,20-dione 21-(dihydrogen phosphate) disodium salt [2392-39-4; 312-93-6]. DEFINITION Dexamethasone Sodium Phosphate contains NLT 97.0% and NMT 102.0% of C22H28FNa2O8P, calculated on the water-free and alcohol-free basis. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Solution A: Mix 3.1 g of boric acid and 500 mL of water in a 1-L volumetric flask, add 21 mL of 1 N sodium hydroxide and 10 mL of 0.1 M magnesium chloride, and dilute with water to volume. Solution B: Transfer 95 5 mg of alkaline phosphatase enzyme to a 50-mL volumetric flask, and dissolve by adding Solution A to volume. Prepare this solution fresh daily. Standard solution: 3 mg/mL of USP Dexamethasone RS in ethyl acetate[NOTESonication may be required to ensure dissolution.] Sample solution: Weigh 20 mg of Dexamethasone Sodium Phosphate in a 15-mL centrifuge tube. Add 5.0 mL of Solution B, shake vigorously, and allow to stand for 30 min. Add 5.0 mL of ethyl acetate, shake vigorously, centrifuge, and use the upper, ethyl acetate layer. Application volume: 10 L Developing solvent system: Chloroform, methanol, and water (180:15:1) Analysis Samples: Standard solution and Sample solution Develop the chromatogram in Developing solvent system, to a distance of three-fourths of the length of the plate. Airdry the plate and observe under short-wavelength UV light. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. B. IDENTIFICATION TESTSGENERAL, Phosphate 191 and Sodium 191: The residue from its ignition meets the requirements. ASSAY PROCEDURE Buffer solution: 7.0 g/L of ammonium acetate in water. Adjust with glacial acetic acid to a pH of 4.00 0.05.
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mL of 2 N sulfuric acid, by warming if necessary. Add 1 mL each of Phosphate reagent A and Phosphate reagent B, dilute with water to 25 mL, and allow to stand at room temperature for 30 min. Spectrometric conditions Absorbance wavelength: 730 nm Cell: 1 cm Blank: Water Analysis Samples: Standard solution and Sample solution Acceptance criteria: The absorbance of the Sample solution is NMT that of the Standard solution (NMT 1.0%). Organic Impurities PROCEDURE 1: ALCOHOL DETERMINATION, Method II 611: Proceed as directed in the chapter, except use column packing S8 and use the following modifications. Internal standard solution: Isopropyl alcohol in water (1 in 100) Standard stock solution: Alcohol in water (1 in 50). Determine the specific gravity at 25 (see Specific Gravity 841), and obtain the concentration (mg/mL) of C2H5OH by reference to the Alcoholometric Table in the section Reference Tables. Standard solution: Transfer 4 mL of Standard stock solution and 5 mL of Internal standard solution to a 10-mL volumetric flask, and dilute with water to volume. Sample stock solution: 100 mg/mL of Dexamethasone Sodium Phosphate in Internal standard solution Sample solution: 10 mg/mL in water from Sample stock solution Injection size: 2 L Analysis: Calculate the percentage of alcohol in the Dexamethasone Sodium Phosphate taken: Result = (RU/RS) (CS/CU) 100 = response ratio of alcohol to the Internal standard from the Sample solution = response ratio of alcohol to the Internal standard RS from the Standard solution CS = concentration of alcohol in the Standard solution (mg/mL) = concentration of dexamethasone sodium CU phosphate in the Sample solution (mg/mL) Acceptance criteria: NMT 8.0% PROCEDURE 2: LIMIT OF FREE DEXAMETHASONE Solution A: 7.5 mL/L of triethylamine. Adjust by the addition of phosphoric acid to a pH of 5.4. Mobile phase: Methanol and Solution A (26:74) Standard solution: 50 g/mL of USP Dexamethasone Phosphate RS and 0.5 g/mL of USP Dexamethasone RS in Mobile phase Sample solution: 50 g/mL of Dexamethasone Sodium Phosphate in Mobile phase System suitability solution: 50 g/mL of USP Dexamethasone Phosphate RS and 20 g/mL of USP Dexamethasone RS in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.5-mm 25-cm; 5-m packing L11 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: System suitability solution Suitability requirements Resolution: NLT 1.8 between dexamethasone phosphate and dexamethasone RU
Solution A: Methanol, water, and Buffer solution (7:7:6) Solution B: Methanol and Buffer solution (7:3) Mobile phase: See the gradient table below.
Time (min) 0 3.5 24 35 60 60.1 65 Solution A (%) 90 90 60 5 5 90 90 Solution B (%) 10 10 40 95 95 10 10
Standard solution: 0.92 mg/mL of USP Dexamethasone Phosphate RS in Solution A Sample solution: 1.0 mg/mL of Dexamethasone Sodium Phosphate in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L7 Temperature: 40 Flow rate: 1 mL/min Injection size: 15 L System suitability Sample: Standard solution and Sample solution Suitability requirements Resolution: NLT 1.0 between dexamethasone phosphate and the nearest impurity eluting after it, Sample solution Relative standard deviation: NMT 2.0%, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H28FNa2O8P taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Dexamethasone Phosphate RS in the Standard solution (mg/mL) CU = concentration of the Sample solution (mg/mL) = molecular weight of dexamethasone sodium Mr1 phosphate, 516.41 = molecular weight of dexamethasone phosphate, Mr2 472.45 Acceptance criteria: 97.0%102.0% rU rS CS IMPURITIES Inorganic Impurities LIMIT OF PHOSPHATE IONS Phosphate reagent A: 50 mg/mL of ammonium molybdate in 1 N sulfuric acid Phosphate reagent B: 3.5 mg/mL of pmethylaminophenol sulfate and 200 mg/mL of sodium bisulfite Standard stock solution: 0.1433 mg/mL of dried monobasic potassium phosphate, KH2PO4. This solution contains the equivalent of 0.10 mg/mL of phosphate (PO4). Standard solution: Mix 5.0 mL of Standard stock solution, 10 mL of water, and 5 mL of 2 N sulfuric acid. Add 1 mL each of Phosphate reagent A and Phosphate reagent B, dilute with water to 25 mL, and allow to stand at room temperature for 30 min. Sample solution: Dissolve 50 mg of Dexamethasone Sodium Phosphate in a mixture of 10 mL of water and 5
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Column efficiency: NLT 900 theoretical plates Tailing factor: NMT 1.6 for the analyte peak Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H29FO5 in the portion of Dexamethasone Sodium Phosphate taken: Result = (rU/rS) (CS/CU) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (g/mL) = concentration of Dexamethasone Sodium CU Phosphate in the Sample solution (g/mL) Acceptance criteria: NMT 1.0% PROCEDURE 3 Acetate buffer: 7 g/L of ammonium acetate in water. Adjust with glacial acetic acid to a pH of 4.0. Solution A: Methanol, water, and Acetate buffer (7:7:6) Solution B: Methanol and Acetate buffer (7:3) Mobile phase: See the gradient table below.
Time (min) 0 3.5 23.5 34.5 59.5 60 Solution A (%) 90 90 60 5 5 90 Solution B (%) 10 10 40 95 95 10

and of alcohol content determined as directed in the test for Alcohol Determination, does not exceed 16.0%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dexamethasone RS USP Dexamethasone Phosphate RS
rU rS CS
Dexamethasone Sodium Phosphate Inhalation Aerosol

(Comment on this Monograph)id=m23460=Dexamethasone Sodium Phosphate Inhalation Aerosol=D-Monos.pdf) DEFINITION Dexamethasone Sodium Phosphate Inhalation Aerosol is a suspension, in suitable propellants and alcohol, in a pressurized container, of dexamethasone sodium phosphate (C22H28FNa2O8P) equivalent to NLT 90.0% and NMT 110.0% of the labeled amount of dexamethasone phosphate (C22H30FO8P). IDENTIFICATION PROCEDURE Standard solution: 300 g/mL of USP Dexamethasone RS in methylene chloride Sample solution: Prepare a pH 9.0 buffer solution by dissolving 3.1 g of boric acid, 203 mg of magnesium chloride, and 860 mg of sodium hydroxide in water to make 1000 mL. Dissolve 50 mg of alkaline phosphatase enzyme in 50 mL of the pH 9.0 buffer solution, and transfer 5 mL of the resulting solution to a glass-stoppered, 50-mL tube containing 5 mL of the Sample solution prepared as directed in the Assay. Incubate at 37 for 45 min, add 25 mL of methylene chloride, and shake for 2 min. Evaporate 15 mL of the methylene chloride extract on a steam bath to dryness, and dissolve the residue in 1 mL of methylene chloride. The methylene chloride extract so obtained responds to: Adsorbent: 20- 20-cm, thin-layer chromatographic plate coated with a 0.25-mm layer of chromatographic silica gel Application volume: 5 L Spray reagent: Dilute sulfuric acid (1 in 2) Developing solvent system: Chloroform, acetone, and water (50:50:1) Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram in a tank completely lined with a strip of filter paper, using developing solvent system, until the solvent front has moved three-fourths of the length of the plate. Spray the plate with Spray reagent, and heat at 105 until brown or black spots appear. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Standard solution: Transfer 40 mg of USP Dexamethasone RS to a 50-mL volumetric flask, and dilute with alcohol to volume. Transfer 5.0 mL of this solution to a 500-mL volumetric flask, and dilute with 0.1 N sulfuric acid to volume to obtain a solution having a known concentration of 8 g/mL. Sample solution: Weigh a filled Inhalation Aerosol container, and record the weight (W1). Place the container in a dry ice-acetone bath, and cool for 60 min. Remove the container from the bath, and carefully remove the valve
Sample solution: 1 mg/mL of Dexamethasone Sodium Phosphate in Solution A Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4.6-mm 25-cm; packing L7 Flow rate: 1 mL/min Column temperature: 40 Injection size: 15 L System suitability Sample: Sample solution Suitability requirements Resolution: NLT 1.0 between the major peak and the nearest impurity Relative standard deviation: NMT 4.0% Analysis Sample: Sample solution Calculate the percentage of each impurity in the portion of Dexamethasone Sodium Phosphate taken: Result = (ri/rT) 100 = peak response for each impurity ri = sum of the responses of all peaks rT Acceptance criteria Individual impurity: NMT 1.0% Total impurities: NMT 2.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +74 to +82, calculated on the water-free and alcohol-free basis Sample solution: 10 mg/mL PH 791: 7.510.5, in a solution (1 in 100) WATER DETERMINATION, Method I 921: Determine the water content. The sum of the percentages of water content,
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Spectrometric conditions Cell: 1 cm Analytical wavelength: 239 nm Blank: 0.1 N sulfuric acid Analysis Samples: Standard solution and Sample solution Calculate the g of C22H30FO8P contained in the minimum dose: Result = (AU/AS) (CS N) (Mr1/Mr2) 10 AU AS CS N Mr1 Mr2 = absorbances of the solutions from the Sample solution = absorbances of the solutions from the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (g/mL) = number of sprays discharged to obtain the minimum recommended dose = molecular weight of dexamethasone phosphate, 472.45 = molecular weight of dexamethasone, 392.47
with wire cutters, taking precautions to save all pieces of the valve and cap. With the aid of four 5-mL portions of 0.1 N sulfuric acid, transfer the contents of the container to a beaker previously chilled in the bath. Dry the rinsed empty container and all of its parts in an oven at 105 for 2 h, cool, and weigh (W2). Allow the contents of the beaker to warm to room temperature. After the bulk of the propellant has evaporated, quantitatively transfer the contents of the beaker, with the aid of several mL of 0.1 N sulfuric acid, to a 200-mL volumetric flask, and dilute with 0.1 N sulfuric acid to volume. Transfer 20 mL of this solution to a centrifuge tube, add 10 mL of methylene chloride, shake vigorously for 1 min, and centrifuge. Pipet 10 mL of the clear supernatant into a 100-mL volumetric flask, and dilute with 0.1 N sulfuric acid to volume. Spectrometric conditions Cell: 1 cm Analytical wavelength: 239 nm Blank: 0.1 N sulfuric acid Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30FO8P in each g of Inhalation Aerosol taken: Result = (AU/AS) (CS/L) (Mr1/Mr2) 2 100 = absorbances of the Sample solution = absorbances of the Standard solution = concentration of USP Dexamethasone RS in the Standard solution (g/mL) L = label claim of Dexamethasone Phosphate in the container (mg) Mr1 = molecular weight of dexamethasone sodium phosphate, 472.45 Mr2 = molecular weight of dexamethasone, 392.47 Acceptance criteria: 90.0%110.0% OTHER COMPONENTS ALCOHOL DETERMINATION, Method II 611: C2H5OH 1.7%2.3% of AU AS CS
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, pressurized containers, and avoid exposure to excessive heat. USP REFERENCE STANDARDS 11 USP Dexamethasone RS
Dexamethasone Sodium Phosphate Cream

(Comment on this Monograph)id=m23480=Dexamethasone Sodium Phosphate Cream=D-Monos.pdf) DEFINITION Dexamethasone Sodium Phosphate Cream contains an amount of dexamethasone sodium phosphate (C22H28FNa2O8P) equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of dexamethasone phosphate (C22H30FO8P). IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 300 g/mL of USP Dexamethasone RS in methylene chloride Solution A: Mix 3.1 g of boric acid, 203 mg of magnesium chloride, and 860 mg of sodium hydroxide in water to make 1000 mL. Solution B: 1 mg/mL of alkaline phosphatase enzyme in Solution A Sample solution: Transfer 5 mL of Solution B to a glassstoppered, 50-mL tube containing 5 mL of the Sample solution, prepared as directed in the Assay. Incubate at 37 for 45 min, then add 25 mL of methylene chloride, and shake for 2 min. Evaporate 15 mL of the methylene chloride extract on a steam bath to dryness, and dissolve the residue in 1 mL of methylene chloride. Application volume: 5 L Spray reagent: Dilute sulfuric acid (1 in 2) Developing solvent system: Chloroform, acetone, and water (50:50:1)
PERFORMANCE TESTS DELIVERED DOSE UNIFORMITY OVER THE ENTIRE CONTENTS: Meets the requirements for Metered-Dose Inhalers under Aerosols, Nasal Sprays, Metered-Dose Inhalers, and Dry Powder Inhalers 601. Procedure for dose uniformity Standard stock solution: 1 mg/mL of USP Dexamethasone RS in alcohol Standard solution: 10 g/mL of Standard stock solution, in 0.1 N sulfuric acid Sample solution: Discharge the minimum recommended dose into the sampling apparatus, and detach the inhaler as directed. Rinse the apparatus (filter and interior) with two 5.0-mL portions of 0.1 N sulfuric acid, and transfer the resulting solutions quantitatively to a 50-mL centrifuge tube containing 15 mL of methylene chloride that was previously chilled in a dry ice-acetone bath for a few min. Insert the stopper in the centrifuge tube, and shake cautiously, releasing the pressure occasionally. Allow the phases to separate, and equilibrate to room temperature. The aqueous phase is the Sample solution. Analysis: Transfer the Sample solution and 10.0 mL of the Standard solution into separate flasks. Add 2.0 mL of 0.1 N sulfuric acid to each, and swirl.
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Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram using the Developing solvent system, until the solvent front has moved about three-fourths of the length of the plate. Spray the plate with Spray reagent, and heat at 105 until brown or black spots appear: the RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Solution A: 0.29 g/L of dibasic sodium phosphate in a mixture of alcohol and water (55:45) Solution B: 6.9 g/L of monobasic sodium phosphate Mobile phase: Methanol and Solution B (52:48) Standard solution: 30 g/mL of USP Dexamethasone Phosphate RS in Solution A. Prepare this solution fresh. Sample solution: Cream, equivalent to 3 mg of dexamethasone phosphate, in a 150-mL beaker. Add 65 mL of Solution A, and heat just to boiling. Pour the contents of the beaker into a 125-mL separator containing 45 mL of isooctane. After shaking for 1 min, decant the lower layer into a 100-mL volumetric flask with the aid of a glass funnel. Rinse the 150-mL beaker with two 15-mL portions of Solution A, extracting the remaining isooctane in the separator with each portion and decanting the lower layer from each extraction into the 100-mL volumetric flask. Dilute with Solution A to volume. Pass through a membrane filter before injecting. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.5 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention time for dexamethasone phosphate is about 8.5 min.] Suitability requirements Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30FO8P in the Cream taken: Result = (rU/rS) (CS/CU) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Dexamethasone Phosphate RS in the Standard solution (g/mL) CU = concentration of the dexamethasone phosphate in the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% SPECIFIC TESTS MICROBIAL ENUMERATION TESTS 61 AND TESTS FOR SPECIFIED MICROORGANISMS 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa MINIMUM FILL 755: Meets the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible tubes or tight containers. USP REFERENCE STANDARDS 11 USP Dexamethasone RS USP Dexamethasone Phosphate RS rU rS CS
Dexamethasone Sodium Phosphate Injection

(Comment on this Monograph)id=m23490=Dexamethasone Sodium Phosphate Injection=D-Monos.pdf) DEFINITION Dexamethasone Sodium Phosphate Injection is a sterile solution of Dexamethasone Sodium Phosphate in Water for Injection. It contains NLT 90.0% and NMT 115.0% of the labeled amount of dexamethasone phosphate (C22H30FO8P), present as the disodium salt. IDENTIFICATION THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Standard solution: 300 g/mL of USP Dexamethasone RS in methylene chloride Solution A: Mix 3.1 g of boric acid and 500 mL of water in a 1-L volumetric flask. Add 21 mL of 1 N sodium hydroxide and 10 mL of 0.1 M magnesium chloride, and dilute with water to volume. Solution B: 1 mg/mL of alkaline phosphatase enzyme in Solution A Sample solution: Injection, equivalent to 10 mg of dexamethasone phosphate in 100-mL of water. Pipet 5 mL of this solution into a 125-mL separator, and wash with two 10-mL portions of water-washed methylene chloride, discarding the washings. Transfer the solution into a glassstoppered, 50-mL tube, and add 5 mL of Solution B. Allow to stand at 37 for 45 min, and extract with 25 mL of methylene chloride. Evaporate 15 mL of the methylene chloride extract on a steam bath to dryness, and dissolve the residue in 1 mL of methylene chloride. Application volume: 5 L Spray reagent: Dilute sulfuric acid (1 in 2) Developing solvent system: Chloroform, acetone, and water (50:50:1) Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram using the Developing solvent system, until the solvent front has moved about three-fourths of the length of the plate. Spray the plate with Spray reagent, and heat at 105 until brown or black spots appear: the RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: 0.01 M monobasic potassium phosphate in a mixture of methanol and water (1:1) Standard solution: 0.08 mg/mL of USP Dexamethasone Phosphate RS in Mobile phase [NOTEPrepare this solution at the time of use.] Sample solution: 0.08 mg/mL of dexamethasone phosphate from Injection, in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Flow rate: 1.6 mL/min Injection size: 20 L System suitability Sample: Standard solution [NOTEThe relative retention time of dexamethasone phosphate is about 5 min.]
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Spray reagent: Dilute sulfuric acid (1 in 2) Analysis Samples: Standard solution and Sample solution Allow the spots to dry, and develop the chromatogram in a tank completely lined with a strip of filter paper, using solvent system, until the solvent front has moved about three-fourths of the length of the plate. Spray the plate with Spray reagent, and heat at 105 until brown or black spots appear. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Solution A: Dissolve 0.29 g of dibasic sodium phosphate in 450 mL of water, and add 550 mL of alcohol. Solution B: In a 1-L volumetric flask, dissolve 6.9 g of monobasic sodium phosphate in 500 mL of water, and dilute with water to volume. Mobile phase: Degassed solution of methanol and Solution B (52:48) which, at ambient temperature and at a flow rate of 1.5 mL/min, gives a retention time of about 8.5 min for dexamethasone phosphate. Standard solution: 30 g/mL of USP Dexamethasone Phosphate RS in Solution A. Prepare this solution fresh. Sample solution: Ophthalmic ointment, nominally equivalent to 3 mg of dexamethasone phosphate, in a 150mL beaker. Add 65 mL of Solution A, and heat just to boiling. Pour the contents of the beaker into a 125-mL separator containing 45 mL of isooctane. After shaking for 1 min, decant the lower layer into a 100-mL volumetric flask with the aid of a glass funnel. Rinse the 150-mL beaker with two 15-mL portions of Solution A, extracting the remaining isooctane in the separator with each portion and decanting the lower layer from each extraction into the 100-mL volumetric flask. Dilute with Solution A to volume and pass through a membrane filter. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Injection size: 20 L System suitability Sample: Standard solution (five replicate injections) Suitability requirements Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30FO8P in the portion of Ophthalmic ointment taken: Result = (rU/rS) (CS/CU) 100 = peak responses at equivalent retention times from the Sample solution = peak responses at equivalent retention times rS from the Standard solution = concentration of USP Dexamethasone Phosphate CS RS in the Standard solution (g/mL) = nominal concentration of dexamethasone CU phosphate in the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% rU PERFORMANCE TESTS MINIMUM FILL 755: Meets the requirements SPECIFIC TESTS METAL PARTICLES IN OPHTHALMIC OINTMENTS 751: the requirements Meets
Suitability requirements Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30FO8P in each mL of the Injection taken: Result = (rU/rS) (CS/CU) 100 = peak responses from the Sample solution = peak responses from the Standard solution = concentration of USP Dexamethasone Phosphate RS in the Standard solution (mg/mL) = nominal concentration of the Injection in the CU Sample solution (mg/mL) Acceptance criteria: 90.0%115.0% rU rS CS SPECIFIC TESTS PH 791: 7.08.5 OTHER REQUIREMENTS: Meets the requirements under Injections 1 BACTERIAL ENDOTOXINS TEST 85: The Injection contains NMT 31.3 USP Endotoxin Units/mg of dexamethasone phosphate. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose or multiple-dose containers, preferably of Type I glass, protected from light. USP REFERENCE STANDARDS 11 USP Dexamethasone RS USP Dexamethasone Phosphate RS USP Endotoxin RS
Dexamethasone Sodium Phosphate Ophthalmic Ointment

(Comment on this Monograph)id=m23500=Dexamethasone Sodium Phosphate Ophthalmic Ointment=D-Monos.pdf) DEFINITION Dexamethasone Sodium Phosphate Ophthalmic Ointment is a sterile ointment containing an amount of dexamethasone sodium phosphate (C22H28FNa2O8P) equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of dexamethasone phosphate (C22H30FO8P). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY Buffer: Dissolve 3.1 g of boric acid, 203 mg of magnesium chloride, and 860 mg of sodium hydroxide in water to make 1000 mL. Enzyme solution: 1 mg/mL alkaline phosphatase enzyme in Buffer Standard solution: 300 g/mL of USP Dexamethasone RS in methylene chloride Sample solution: Transfer 5 mL of the Sample solution obtained in the Assay to a 50-mL glass stoppered tube. Add 5 mL of the Enzyme solution and incubate at 37 for 45 min, then add 25 mL of methylene chloride, and shake for 2 min. Evaporate 15 mL of the methylene extract to dryness. Dissolve the residue in 1 mL of methylene chloride. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 20- 20-cm, thin-layer chromatographic plate coated with a 0.25-mm layer of chromatographic silica gel Application volume: 5 L Developing solvent system: Chloroform, acetone, and water (50:50:1)
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STERILITY TESTS 71: Meets the requirements
Official Monographs / Dexbrompheniramine 55

Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30-cm; packing L1 Injection size: 20 L System suitability Sample: Standard solution (five replicate injections) Suitability requirements Relative standard deviation: NMT 1.5% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C22H30FO8P in each mL of the Ophthalmic Solution taken: Result = (rU/rS) (CS/CU) 100 = peak response at equivalent retention times from the Sample solution rS = peak response at equivalent retention times from the Standard solution CS = concentration of USP Dexamethasone Phosphate RS in the Standard solution (g/mL) CU = concentration of dexamethasone phosphate in the Sample solution (g/mL) Acceptance criteria: 90.0%115.0% rU SPECIFIC TESTS PH 791: 6.67.8 STERILITY TESTS 71:
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in collapsible ophthalmic ointment tubes. USP REFERENCE STANDARDS 11 USP Dexamethasone RS USP Dexamethasone Phosphate RS
Dexamethasone Sodium Phosphate Ophthalmic Solution

(Comment on this Monograph)id=m23510=Dexamethasone Sodium Phosphate Ophthalmic Solution=D-Monos.pdf) DEFINITION Dexamethasone Sodium Phosphate Ophthalmic Solution is a sterile, aqueous solution of Dexamethasone Sodium Phosphate. It contains an amount of dexamethasone sodium phosphate (C22H28FNa2O8P) equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of dexamethasone phosphate (C22H30FO8P). IDENTIFICATION THIN-LAYER CHROMATOGRAPHY (See Chromatography 621.) Adsorbent: 20- 20-cm, thin-layer chromatographic plate coated with a 0.25-mm layer of chromatographic silica gel Solution A: 3.1 mg/mL of boric acid, 0.203 mg/mL of magnesium chloride, and 0.86 mg/mL of sodium hydroxide in water Solution B: 1 mg/mL of alkaline phosphate enzyme in Solution A Standard solution: 300 g/mL of USP Dexamethasone RS in methylene chloride Sample solution: Transfer 5 mL of Solution B to a glass stoppered, 50-mL tube containing 5 mL of Assay Sample solution. Incubate at 37 for 45 min, then add 25 mL of methylene chloride, and shake for 2 min. Evaporate 15 mL of the methylene chloride extract on a steam bath to dryness, and dissolve the residue in 1 mL of methylene chloride. Application volume: 5 L Spray reagent: Dilute sulfuric acid (1 in 2) Developing solvent system: Chloroform, acetone, and water (50:50:1), in a tank that is completely lined with a strip of filter paper Analysis Samples: Standard solution and Sample solution Apply Sample solution and Standard solution to plate, allow spots to dry, and develop until the solvent front has moved about three-fourths of the length of the plate. Spray the plate with Spray reagent, and heat at 105 until brown or black spots appear. Acceptance criteria: The RF value of the principal spot of the Sample solution corresponds to that of the Standard solution. ASSAY PROCEDURE Mobile phase: Prepare 0.01 M monobasic potassium phosphate in a mixture of methanol and water (1:1), at ambient temperature and at a flow rate of about 1.6 mL/min, gives a retention time of about 5 min for dexamethasone phosphate. Standard solution: [NOTEPrepare this solution at the time of use.] 80 g/mL of USP Dexamethasone Phosphate RS in Mobile phase Sample solution: 80 g/mL of dexamethasone phosphate from Ophthalmic Solution, in Mobile phase
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Dexamethasone RS USP Dexamethasone Phosphate RS
Dexbrompheniramine Maleate
(Comment on this Monograph)id=m23560=Dexbrompheniramine Maleate=DMonos.pdf)
435.32 C16H19BrN2 C4H4O4 2-Pyridinepropanamine, -(4-bromophenyl)-N,N-dimethyl-, (S)-, (Z)-2-butenedioate (1:1); (+)-2-[p-Bromo--[2-(dimethylamino)ethyl]benzyl]pyridine maleate (1:1) [2391-03-9]. DEFINITION Dexbrompheniramine Maleate contains NLT 98.0% and NMT 100.5% of C16H19BrN2 C4H4O4, calculated on the dried basis. IDENTIFICATION A. INFRARED ABSORPTION 197M B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 261 nm Sample solution: 35 g/mL in methanol Absorptivities: Do not differ by more than 3.0%, calculated on the dried basis
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Dexbrompheniramine / Official Monographs
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IDENTIFICATION A. The retention time of the major peak for dexbrompheniramine maleate in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. B. The retention time of the major peak for pseudoephedrine sulfate in the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. C. IDENTIFICATION TESTSGENERAL, Sulfate 191: A solution of it meets the requirements. D. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST Standard solution A: 1.2 mg/mL of USP Dexbrompheniramine Maleate RS in methanol Standard solution B: 9 mg/mL of USP Pseudoephedrine Sulfate RS in methanol Sample solution: Oral Solution, equivalent to 6 mg of dexbrompheniramine maleate, to a separatory funnel, add 0.5 mL of ammonium hydroxide and 5 mL of methylene chloride, shake for 1 min, and allow the layers to separate. Use the clear, lower layer. Chromatographic system (See Chromatography 621, Thin-Layer Chromatography.) Mode: TLC Adsorbent: 0.25-mm layer of chromatographic silica gel mixture Application volume: 5 L Developing solvent system: Methanol, ethyl ether, and ammonium hydroxide (3:16:1) Analysis Samples: Standard solution A, Standard solution B, and Sample solution Allow the spots to dry, and develop the chromatogram until the solvent front has moved about three-fourths of the length of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow the solvent to evaporate. Locate the spots on the plate by examination under short-wavelength UV light. Acceptance criteria: The RF values of the two principal spots of the Sample solution correspond to those of the respective Standard solutions. ASSAY PROCEDURE Mobile phase: Acetonitrile, methanol, tetrahydrofuran, and water (32:8:5:55) Transfer 0.1 mL of phosphoric acid, followed by 0.433 g of sodium lauryl sulfate, to each 100 mL of this mixture. Adjust with ammonium hydroxide to a pH of 3.50 0.05. [NOTEThe pH of the Mobile phase is critical, and may cause differences of 14 min in the retention times of the internal standard and dexbrompheniramine.] Internal standard solution: 0.5 mg/mL of naphazoline hydrochloride in Mobile phase Dexbrompheniramine standard solution: 6000J g/mL, (J being the ratio of the labeled amount, in mg, of dexbrompheniramine maleate to the labeled amount, in mg, of pseudoephedrine sulfate per mL of the Oral Solution) of USP Dexbrompheniramine Maleate RS in Mobile phase Standard solution: Transfer 30 mg of USP Pseudoephedrine Sulfate RS to a 25-mL volumetric flask, add 5.0 mL each of Dexbrompheniramine standard solution and Internal standard solution, dilute with Mobile phase to volume, and mix to obtain a Standard solution having known concentrations of about 1.2J mg/mL of USP Dexbrompheniramine Maleate RS and 1.2 mg/mL of USP Pseudoephedrine Sulfate RS. Sample solution: Using a to contain pipet, transfer a volume of Oral Solution, equivalent to 30 mg of pseudoephedrine sulfate, to a 25-mL volumetric flask. Rinse the pipet with 5 mL of Mobile phase, collecting the rinsing
ASSAY PROCEDURE Sample solution: Dissolve 400 mg of Dexbrompheniramine Maleate in 50 mL of glacial acetic acid. Analysis: Titrate with 0.1 N perchloric acid VS to a green endpoint, using 1 drop of crystal violet TS, as an indicator. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 21.77 mg of C16H19BrN2 C4H4O4. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Sample solution: 40 mg/mL of Dexbrompheniramine Maleate in methylene chloride Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 4-mm 1.2-m; glass column containing 3% phase G3 on support S1AB Temperature Column: 190 Injection port: 250 Detector: 250 Carrier gas: Dry helium, flowing at a rate adjusted to obtain a retention time of 6 or 7 min for the main peak Injection size: 1 L Run time: NLT twice the retention time of the dexbrompheniramine peak System suitability Sample: Sample solution Suitability requirements Tailing factor: NMT 1.8 Analysis Sample: Sample solution Measure the areas of the peaks. Acceptance criteria: The total relative area of all extraneous peaks (except that of the solvent peak and the maleic acid, if observed) does not exceed 2.0%. SPECIFIC TESTS LOSS ON DRYING 731: Dry it at 65 for 4 h: it loses NMT 0.5% of its weight. OPTICAL ROTATION, Specific Rotation 781S: +35.0 to +38.5 Sample solution: 50 mg/mL, in dimethylformamide ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Dexbrompheniramine Maleate RS
Dexbrompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution

(Comment on this Monograph)id=m23570=Dexbrompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution=D-Monos.pdf) DEFINITION Dexbrompheniramine Maleate and Pseudoephedrine Sulfate Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amounts of dexbrompheniramine maleate (C10H15BrN2 C4H4O4) and pseudoephedrine sulfate [(C10H15NO)2 H2SO4].
USP 32
in the volumetric flask. Add 5.0 mL of Internal standard solution, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 254 nm Column: 4-mm 30 -cm; packing L11 Flow rate: 1.5 mL/min Injection size: 10 L System suitability Sample: Standard solution [NOTEThe relative retention times for pseudoephedrine, naphazoline, and dexbrompheniramine are about 1.0, 1.5, and 2.5, respectively.] Suitability requirements Resolution: NLT 3.0 between the pseudoephedrine and naphazoline peaks; NLT 3.0 between the dexbrompheniramine and naphazoline peaks Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C10H15BrN2 C4H4O4 and pseudoephedrine sulfate (C10H15NO)2 H2SO4 in the portion of Oral Solution taken: Result = (RU/RS) (CS/CU) 100 = peak response ratio of the corresponding analyte to the internal standard from the Sample solution = peak response ratio of the corresponding analyte RS to the internal standard from the Standard solution = concentration of the appropriate USP Reference CS Standard in the Standard solution (mg/mL) = nominal concentration of dexbrompheniramine CU maleate or pseudoephedrine sulfate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% RU PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: For Oral Solution packaged in single-unit containers: meets the requirements DELIVERABLE VOLUME 698: For Oral Solution packaged in multiple-unit containers: meets the requirements ADDITIONAL REQUIREMENTS USP REFERENCE STANDARDS 11 USP Dexbrompheniramine Maleate RS USP Pseudoephedrine Sulfate RS
Official Monographs / Dexchlorpheniramine 57

DEFINITION Dexchlorpheniramine Maleate, dried at 65 for 4 h, contains NLT 98.0% and NMT 100.5% of C16H19ClN2 C4H4O4. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Sample solution: 40 g/mL ASSAY PROCEDURE Sample: 400 mg of previously dried Dexchlorpheniramine Maleate Analysis: Dissolve in glacial acetic acid, add 1 drop of crystal violet TS, and titrate with 0.1 N perchloric acid VS to a green endpoint. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 19.54 mg of C16H19ClN2 C4H4O4. Acceptance criteria: 98.0%100.5% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.2% Organic Impurities PROCEDURE Sample solution: 40 mg/mL of Dexchlorpheniramine Maleate in methylene chloride Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.2-m 4-mm glass column containing 3% phase G3 on support S1AB Temperature Column: 190 Injection port: 250 Detector: 250 Flow rate: Adjusted to obtain a retention time of 45 min for the main peak Injection size: 1 L Carrier gas: Helium System suitability Sample: Sample solution Suitability requirements Tailing factor: NMT 1.8, for the dexchlorpheniramine maleate peak Analysis Sample: Sample solution Record the chromatogram for a total time not less than twice the retention time of the dexchlorpheniramine peak, and measure the areas of the peaks. Acceptance criteria: The total relative area of all extraneous peaks (except that of the solvent peak and maleic acid, if observed) does not exceed 2.0%. SPECIFIC TESTS MELTING RANGE OR TEMPERATURE, Class I 741: 110115 OPTICAL ROTATION, Specific Rotation 781S: +39.5 to +43.0 Sample solution: 50 mg/mL, in dimethylformamide PH 791: 4.05.0, in a solution (1 in 100) LOSS ON DRYING 731: Dry it at 65 for 4 h: it loses NMT 0.5% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Dexchlorpheniramine Maleate RS
Dexchlorpheniramine Maleate
(Comment on this Monograph)id=m23610=Dexchlorpheniramine Maleate=DMonos.pdf)
390.86 C16H19ClN2 C4H4O4 2-Pyridinepropanamine, -(4-chlorophenyl)-N,N-dimethyl-, (S)-, (Z)-2-butenedioate (1:1); (+)-2-[p-Chloro--[2-(dimethylamino)ethyl]benzyl]pyridine maleate (1:1) [2438-32-6].
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Dexchlorpheniramine / Official Monographs

AU AS CS
USP 32
= absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Dexchlorpheniramine Maleate RS in the Standard solution (g/mL) = nominal concentration of dexchlorpheniramine CU maleate in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% OTHER COMPONENTS ALCOHOL DETERMINATION 611: 5.0%7.0%
Dexchlorpheniramine Maleate Oral Solution

(Comment on this Monograph)id=m23640=Dexchlorpheniramine Maleate Oral Solution=D-Monos.pdf) DEFINITION Dexchlorpheniramine Maleate Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of dexchlorpheniramine maleate (C16H19ClN2 C4H4O4). IDENTIFICATION A. Evaporate the remaining extract from the Assay on a steam bath to a small volume, then transfer it to a smaller, more suitable vessel, and evaporate just to the point where hexane vapors are no longer perceptible. Transfer the oily residue, with the aid of four 3-mL portions of dimethylformamide, to a suitable glass-stoppered graduated cylinder, and dilute with dimethylformamide to 15.0 mL. The optical rotation of the solution so obtained, in a 100mm tube, after correcting for the blank, is between +0.06 and +0.11 (distinction from chlorpheniramine maleate). B. ULTRAVIOLET ABSORPTION 197U: Sample solution compared to Standard solution from Assay ASSAY PROCEDURE Standard stock solution: 0.4 mg/mL of USP Dexchlorpheniramine Maleate RS Standard solution: 40 g/mL of USP Dexchlorpheniramine Maleate RS in Standard solution, by transferring 10.0 mL of Standard stock solution to a separator, adjust with 1 N sodium hydroxide to a pH of 11, and cool. Extract with two 50-mL portions of solvent hexane, shaking each portion for 2 min before separating the phases, and combining the hexane extracts in a second separator. Extract the hexane solution with two 40-mL portions of dilute hydrochloric acid (1 in 120), combine the acid extracts in a 100-mL volumetric flask, and add dilute hydrochloric acid (1 in 120) to volume. Filter the solution into a glass-stoppered conical flask, discarding the first few mL of the filtrate. Sample solution: Equivalent to 40 mg of dexchlorpheniramine maleate from Oral Solution, in a 250mL separator, using a pipet calibrated to contain the required volume. Rinse the pipet with small portions of water, add the rinsings to the separator, adjust with 1 N sodium hydroxide to a pH of 11, and cool. Extract with five 70-mL portions of solvent hexane, combine the hexane extracts in a 500-mL separator, and wash the hexane solution with two 10-mL portions of sodium hydroxide solution (1 in 250). Extract the combined alkaline washings with two 20-mL portions of solvent hexane, and add these extracts to the bulk of the alkali-washed hexane solution. Filter the hexane solution through a pledget of cotton that previously has been saturated with solvent hexane into a 500-mL volumetric flask, rinse the separator with portions of solvent hexane, and pass the rinsings through the filter to add to volume. Transfer 50.0 mL of this solution to a separator (retain the remaining extract for Identification test A), and proceed as directed for Standard solution, beginning with Extract the hexane solution. Spectrometric conditions Analytical wavelength: 264 nm Cell: 1 cm Blank: Diluted hydrochloric acid (8.33 mg/mL) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in each mL of the Oral Solution taken: Result = (AU/AS) (CS/CU) 100
ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Dexchlorpheniramine Maleate RS
Dexchlorpheniramine Maleate Tablets

(Comment on this Monograph)id=m23650=Dexchlorpheniramine Maleate Tablets=D-Monos.pdf) DEFINITION Dexchlorpheniramine Maleate Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of C16H19ClN2 C4H4O4. IDENTIFICATION A. IDENTIFICATIONORGANIC NITROGENOUS BASES 181 Tablets meet the requirements B. PROCEDURE Sample: Finely powdered Tablets equivalent to 150 mg of dexchlorpheniramine maleate Analysis: Shake a quantity of Sample with 100 mL of 1 N acetic acid for 10 min, filter through a sintered-glass funnel into a suitable vessel, adjust the filtrate with sodium hydroxide solution (1 in 10) to a pH of 11, and extract the solution with six 100-mL portions of solvent hexane, filtering each hexane extract using suitable means to separate the hexane layer from the aqueous layer. Concentrate the combined extracts on a steam bath to a small volume, transfer to a smaller, more suitable vessel, and evaporate just to the point where hexane vapors are no longer perceptible. Transfer the oily residue, with the aid of four 3-mL portions of dimethylformamide, to a suitable glass-stoppered graduated cylinder, dilute with dimethylformamide to 15.0 mL, and centrifuge if necessary. Acceptance criteria: The optical rotation of the solution so obtained in a 100-mm tube after correcting for the blank is between +0.24 and +0.35 (distinction from chlorpheniramine maleate). ASSAY PROCEDURE Standard stock solution: 0.4 mg/mL of USP Dexchlorpheniramine Maleate RS Standard solution: Transfer 10.0 mL of the Standard stock solution to a separator, adjust with 1 N sodium hydroxide to a pH of 11, and cool. Extract with two 50-mL portions of solvent hexane, shaking each portion for 2 min before separating the phases, and combining the hexane extracts in a second separator. Extract the hexane solution with two 40-mL portions of dilute hydrochloric acid (1 in 120), combine the acid extracts in a 100-mL volumetric flask, and add dilute hydrochloric acid (1 in 120) to volume. Filter the solution into a glass-stoppered conical flask, discarding the first few mL of the filtrate. The concentration of USP Dexchlorpheniramine Maleate RS in the Standard solution is 40 g/mL.
USP 32
Sample solution: Transfer an equivalent to 8 mg of dexchlorpheniramine maleate, from finely powdered Tablets (NLT 20), to a 250-mL separator, mix with 50 mL of water for 10 min, adjust with sodium hydroxide solution (1 in 10) to a pH of 11, and cool to room temperature. Extract the mixture with two 75-mL portions of solvent hexane, and combine the extracts in a second separator. Extract the solvent hexane solution with three 50-mL portions of dilute hydrochloric acid (1 in 120), combining the acid extracts in a 200-mL volumetric flask. Add dilute hydrochloric acid (1 in 120) to volume. Spectrometric conditions Analytical wavelength: 264 nm Cell: 1 cm Blank: Dilute hydrochloric acid (1 in 120) Analysis Samples: Standard solution and Sample solution Calculate the percentage of C16H19ClN2 C4H4O4 in the portion of Tablets taken: Result = (AU/AS) (CS/CU) 100 = absorbance of the Sample solution = absorbance of the Standard solution = concentration of USP Dexchlorpheniramine Maleate RS in the Standard solution (g/mL) = nominal concentration of dexchlorpheniramine CU maleate in the Sample solution (g/mL) Acceptance criteria: 90.0%110.0% AU AS CS PERFORMANCE TESTS DISSOLUTION 711 Medium: Water; 500 mL Apparatus 2: 50 rpm Time: 45 min Determine the amount of C16H19ClN2 C4H4O4 dissolved using the following Analysis. Internal standard solution: 90 g/mL of dexbrompheniramine maleate Standard stock solution: 12.5 g/mL of USP Dexchlorpheniramine Maleate RS Standard solution: Pipet 5 mL of this stock solution into a 50-mL centrifuge tube, and add 10.0 mL water and 1.0 mL Internal standard solution. Adjust with sodium hydroxide solution (1 in 2) to a pH of 11 0.1, and add 3.0 mL of chromatographic solvent hexane. Insert the stopper in the tube, shake by mechanical means for 3 min, centrifuge, and use the clear supernatant hexane layer. Sample solution: Pipet 15 mL of a portion of the Sample solution into a 50-mL centrifuge tube, and add 1.0 mL of Internal standard solution. Adjust with sodium hydroxide solution (1 in 2) to a pH of 11 0.1, and add 3.0 mL of chromatographic solvent hexane. Insert the stopper in the tube, shake by mechanical means for 3 min, centrifuge, and use the clear supernatant hexane layer. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m column that contains a packing consisting of 1.2% phase G16 and 0.5% potassium hydroxide on support S1AB
Official Monographs / Dexpanthenol 59

Temperature Column: 205 Injection port: 250 Detector: 250 Flow rate: 60 mL/min Carrier gas: Helium Injection size: 2 L System suitability Sample: Standard solution [NOTEThe relative retention times for dexchlorpheniramine and dexbrompheniramine are about 0.7 and 1.0, respectively.] Suitability requirements Resolution: NLT 1.9 between dexchlorpheniramine and dexbrompheniramine Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity of C16H19ClN2 C4H4O4 dissolved by comparing the peak response ratios. Tolerances: NLT 75% (Q) of the labeled amount of C16H19ClN2 C4H4O4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dexchlorpheniramine Maleate RS
Dexpanthenol
(Comment on this Monograph)id=m23710=Dexpanthenol=DMonos.pdf)
C9H19NO4 205.25 Butanamide, 2,4-dihydroxy-N-(3-hydroxypropyl)-3,3-dimethyl-, (R)-; D-(+)-2,4-Dihydroxy-N-(3-hydroxypropyl)-3,3dimethylbutyramide [81-13-0]. DEFINITION Dexpanthenol contains NLT 98.0% and NMT 102.0% of C9H19NO4, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197F B. To 1 mL of a solution (1 in 10) add 5 mL of 1 N sodium hydroxide and 1 drop of cupric sulfate TS, and shake vigorously: a deep blue color develops. C. To 1 mL of a solution (1 in 100), add 1 mL of 1 N hydrochloric acid, and heat on a steam bath for 30 min. Cool, add 100 mg of hydroxylamine hydrochloride, and add 5 mL of 1 N sodium hydroxide. Allow to stand for 5 min, then adjust with 1 N hydrochloric acid to a pH of 2.53.0,
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and add 1 drop of ferric chloride TS: a purplish red color develops.
USP 32
ASSAY PROCEDURE Solution A: 20.42 g/L of potassium biphthalate in glacial acetic acid. If necessary, warm the mixture on a steam bath to dissolve, observing precautions against absorption of moisture. Cool to room temperature, and dilute to volume. Sample solution: 400 mg of Dexpanthenol in a 300-mL flask fitted to a reflux condenser by means of a standardtaper glass joint, add 50.0 mL of 0.1 N perchloric acid VS, and reflux for 5 h. Cool, observing precautions to prevent atmospheric moisture from entering the condenser, and rinse the condenser with glacial acetic acid, collecting the rinsings in the flask. Add 5 drops of crystal violet TS. Analysis: Titrate the Sample solution with Solution A to a blue-green endpoint. Perform a blank determination (see Titrimetry 541), and note the differences in volumes required. Each mL of the difference in volumes of 0.1 N perchloric acid consumed is equivalent to 20.53 mg of C9H19NO4. Acceptance criteria: 94.5%98.5% OTHER COMPONENTS CONTENT OF PANTOLACTONE Internal standard solution: 100 mg/mL of 2,6dimethylphenol in toluene Standard solution A: 10 mg/mL of USP Pantolactone RS in methylene chloride Standard solution B: Pipet 0.4 mL of Standard solution A into a suitable small vial. Evaporate the solvent by means of a steady stream of dry air, and add 50 L of Internal standard solution. Add 1 mL of a mixture of pyridine, hexamethyldisilazane, and trimethylchlorosilane (9:3:1), immediately insert the stopper, and shake vigorously for 30 s. Sample solution: Transfer 100 mg of Preparation to a suitable small vial, and proceed as directed for Standard solution B, beginning with add 50 L of Internal standard solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 1.8-m 2.0-mm, packed with 5% liquid phase G2 on support S1A. Temperature Column: 170 Injection port: 180 Flow rate: Using a suitable carrier gas, adjust the flow rate so that the derivatized pantolactone elutes in 4 min. Injection size: 0.5 L System suitability Sample: Standard solution B (five injections) [NOTEThe retention time of the derivatized pantolactone is 0.75, relative to that of the derivatized internal standard.] Suitability requirements Resolution: NLT 2.0, between the two peaks Relative standard deviation: NMT 2.0%, of the peak response ratios (RS) Analysis Samples: Standard solution B and Sample solution Inject Standard solution B into the gas chromatograph, record the chromatogram to obtain NLT 40% of maximum recorder response, and measure the peak responses of the derivatized pantolactone and the derivatized internal standard. Similarly, inject the Sample solution, record the chromatogram, and measure the peak responses of the corresponding components.
ASSAY PROCEDURE Solution A: 20.42 mg/mL of potassium biphthalate in glacial acetic acid [NOTEIf necessary, warm the mixture on a steam bath to achieve complete solution, observing precautions against absorption of moisture. Cool to room temperature, and dilute with glacial acetic acid to volume.] Sample solution: Transfer 400 mg of Dexpanthenol to a 300-mL flask fitted to a reflux condenser by means of a standard-taper glass joint, add 50.0 mL of 0.1 N perchloric acid VS, and reflux for 5 h. Cool, observing precautions to prevent atmospheric moisture from entering the condenser, and rinse the condenser with glacial acetic acid, collecting the rinsings in the flask. Analysis: Titrate the Sample solution with Solution A to a blue-green endpoint, using 5 drops of crystal violet TS as the indicator. Perform a blank determination (see Titrimetry 541), and note the differences in volumes required. Each mL of the difference in volumes of 0.1 N perchloric acid consumed is equivalent to 20.53 mg of C9H19NO4. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE: LIMIT OF AMINOPROPANOL Sample solution: Transfer 5 g to a 50-mL flask, and dissolve in 10 mL of water. Analysis: Titrate with 0.1 N sulfuric acid VS to a yellow endpoint, using bromothymol blue TS as the indicator. Each mL of 0.1 N sulfuric acid is equivalent to 7.5 mg of aminopropanol. Acceptance criteria: NMT 1.0% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +29.0 to +31.5 Sample solution: 50 mg/mL REFRACTIVE INDEX 831: 1.4951.502, at 20 WATER DETERMINATION, Method I 921: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dexpanthenol RS
Dexpanthenol Preparation
(Comment on this Monograph)id=m23717=Dexpanthenol Preparation=D-Monos.pdf) DEFINITION Dexpanthenol Preparation contains NLT 94.5% and NMT 98.5% of dexpanthenol (C9H19NO4), and NLT 2.7% and NMT 4.2% of pantolactone, both calculated on the anhydrous basis. IDENTIFICATION A. The IR absorption spectrum of a thin film of it exhibits maxima only at the same wavelengths as that of a similar preparation of USP Dexpanthenol RS, except that there is an additional maximum at 5.6 m due to pantolactone. B. To 1 mL of a solution (1 in 10) add 5 mL of 1 N sodium hydroxide and 1 drop of cupric sulfate TS, and shake vigorously: a deep blue color develops.
USP 32
Calculate the percentage of pantolactone in the portion of Preparation taken: Result = (RU/RS) (CS/CU) 100 = ratios of the peak response due to the pantolactone to that due to the internal standard from the Sample solution = ratios of the peak response due to the RS pantolactone to that due to the internal standard from the Standard solution B = concentration of USP Pantolactone RS in CS Standard solution A (mg/mL) = nominal concentration for the Sample solution CU (mg/mL) Acceptance criteria: 2.7%4.2% RU SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +27.5 to +30.0 Sample solution: 50 mg/mL REFRACTIVE INDEX 831: 1.4951.502, at 20 WATER DETERMINATION, Method I 921 NMT 1.0% RESIDUE ON IGNITION 281 NMT 0.1% LIMIT OF AMINOPROPANOL Sample solution: 500 mg/mL Analysis: Add bromothymol blue TS to the Sample solution. Titrate with 0.1 N sulfuric acid VS to a yellow endpoint. Each mL of 0.1 N sulfuric acid is equivalent to 7.5 mg of aminopropanol. Acceptance criteria: NMT 1.0% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dexpanthenol RS USP Pantolactone RS
Official Monographs / Dextran 61

0.1 N silver nitrate is equivalent to 5.844 mg of sodium chloride: NMT 1.5% of sodium chloride is found. NITROGEN DETERMINATION 461 (where it is labeled as intended for use in the preparation of injectables) Sulfate solution: To 1000 mL of sulfuric acid, add 5 g of anhydrous cupric sulfate and 500 g of potassium sulfate. Dissolve by heating, and store at 60. [NOTEIf storage at 60 is not possible, prepare a smaller quantity of Sulfate solution on the day of use, adjusting proportions accordingly.] Indicator: Dilute a mixture of 20 mL of a 0.1% solution of bromocresol green in alcohol and 4 mL of methyl red TS with water to 100 mL. Analysis Sample: 0.2 g of Dextran 1 in a micro-Kjeldahl flask Add 4 mL of Sulfate solution. Heat until the solution exhibits a clear green color and the sides of the flask are free from carbonaceous material. Cool, cautiously add 30 mL of water, and transfer the solution to a steam distillation unit. Rinse the Kjeldahl flask with three 5-mL portions of water, adding the washings to the solution. Add 15 mL of 45% sodium hydroxide solution, immediately close the distillation apparatus, and start steam distillation immediately. Receive the distillate in 1 mL of Indicator and sufficient water to cover the end of the condensing tube. Upon completion of the distillation, remove the receiving flask, and rinse the end of the condensing tube with a small quantity of water, adding the rinse to the distillate. Titrate the distillate with 0.010 N hydrochloric acid until the color changes from blue to reddish violet. Perform a blank determination, and make any necessary correction. Acceptance criteria: The corrected volume of 0.010 N hydrochloric acid required to change the color does not exceed 0.15 mL (110 ppm of nitrogen). Organic Impurities PROCEDURE: LIMIT OF ALCOHOL AND RELATED IMPURITIES Sample solution: Dissolve without heating 5.0 g in 100 mL of water, and distill the solution, collecting the first 45 mL of the distillate. Dilute the distillate with water to 50.0 mL. Standard solution: To 25.0 mL of the Sample solution, add 0.5 mL of a 2.5% solution of n-propyl alcohol. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m; packed with S3 Temperature Column: 160 Injection port: 240 Detector: 210 Carrier gas: Nitrogen Flow rate: 25 mL/min [NOTEInjector seals may deteriorate after multiple injections of the Standard solution and Sample solution. Inspect the seals before making a series of injections.] Injection size: 1 L Analysis Samples: Sample solution, Standard solution, and a 0.05% solution of n-propyl alcohol and water Acceptance criteria: After corrections for any impurities in the n-propyl alcohol solution and water, the total area of peaks from impurities in the Sample solution does not exceed the area of the n-propyl alcohol solution peak. SPECIFIC TESTS SPECTROPHOTOMETRY AND LIGHT-SCATTERING, Absorbance 851: NMT 0.12 in a 15% solution at 375 nm, water being used as the blank
Dextran 1
(Comment on this Monograph)id=m23750=Dextran 1=DMonos.pdf) DEFINITION Dextran 1 is a low molecular weight fraction of dextran, consisting of a mixture of isomaltooligosaccharides. It is obtained by controlled hydrolysis and fractionation of dextrans produced by fermentation of Leuconostoc mesenteroides (strain NRRL B-512; CIP 78.59, or its sub-strains, for example L. mesenteroides B-512F; NCTC, 10817), in the presence of sucrose. It is a glucose polymer in which the linkages between glucose units are almost exclusively -1,6. Its weight-average molecular weight is about 1000. IDENTIFICATION A. INFRARED ABSORPTION 197K: To 12 mg each of USP Dextran 1 RS and the sample, add 12 drops of water, grind in an agate mortar for 1 2 min, add 300 mg of potassium bromide, and mix to a slurry. [NOTEDo not grind.] Dry under vacuum at 40 for 15 min, and if it is not dry, continue drying for another 15 min. Crush the residue, prepare a disk, and run the IR spectrum with a blank potassium bromide disk in the reference beam. B. It meets the requirements of the test for Molecular weight distribution and Average molecular weight. IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 5 ppm LIMIT OF SODIUM CHLORIDE: 5 g of Dextran 1 in 100 mL of water. Add 0.2 mL of potassium chromate TS, and titrate with 0.1 N silver nitrate VS (see Titrimetry 541). Each mL of
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Dextran / Official Monographs

Isomaltoundecaose Isomaltododecaose Isomaltotridecaose Isomaltotetradecaose Isomaltopentadecaose Isomaltohexadecaose Isomaltoheptadecaose Isomaltooctadecaose Isomaltononadecaose 1800 1962 2124 2286 2448 2610 2772 2934 3096
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OPTICAL ROTATION, Specific Rotation 781S: +148 to +164 at 20 Sample solution: Solution in water, on the dried basis (dry at 70 under vacuum to constant weight), and corrected for the content of sodium chloride MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: The total aerobic microbial count does not exceed 102 cfu/g, determined by plate-count, and the total combined molds and yeasts count does not exceed 10 cfu/g. PH 791: 4.57.0, in a 15% solution LOSS ON DRYING 731: Dry it at 100 to 105 for 5 h: it loses NMT 5.0% of its weight. BACTERIAL ENDOTOXINS TEST 85 (where it is labeled as intended for use in the preparation of injectables): NMT 25.0 USP Endotoxin Units/g MOLECULAR WEIGHT DISTRIBUTION and AVERAGE MOLECULAR WEIGHT Mobile phase: 2.9 mg/mL of sodium chloride Calibration solution: 0.45 mg of isomaltotriose (3 glucose units) and 0.60 mg of sodium chloride/mL System suitability solution: 6.06.5 mg/mL of USP Dextran 1 RS in Mobile phase Sample solution: 6.06.5 mg/mL solution of Dextran 1 in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Differential refractive index Column: Two 10-mm 30-cm columns in series; packing L54 Temperature: 2025 Flow rate: 0.070.08 mL/min, maintained constant to 1% Injection size: 100 L Analysis Samples: Calibration solution, System suitability solution, and Sample solution Inject the Calibration solution, record the chromatogram, and note the retention times of the peaks. Separately inject the System suitability solution and the Sample solution, and record the chromatograms. Using the retention times in the chromatogram of the Calibration solution, identify the peaks due to isomaltotriose and sodium chloride in the chromatograms of the System suitability solution and the Sample solution. Disregard the peak due to sodium chloride in the System suitability solution and the Sample solution. Calculate the weight-average molecular weight, MW: Result = Wi Mi = weight proportion of oligosaccharide i Wi Mi = molecular weight of oligosaccharide i Use the following molecular weight values for calculation.
Glucose Isomaltose Isomaltotriose Isomaltotetraose Isomaltopentaose Isomaltohexaose Isomaltoheptaose Isomaltooctaose Isomaltononaose Isomaltodecaose 180 342 504 666 828 990 1152 1314 1476 1638
Calculate the amounts of the fractions with fewer than 3 and with more than 9 glucose units for the System suitability solution and the Sample solution. Acceptance criteria: The Mw and the amounts of the fractions obtained for the System suitability solution are within the values stated in the data sheet that accompanies USP Dextran 1 RS. The Mw of Dextran 1 is 8501150. The fraction with fewer than 3 units of glucose is less than 15%, and the fraction with more than 9 units of glucose is less than 20%. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Store in well-closed containers at a temperature between 4 and 30. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Dextran 1 RS USP Endotoxin RS
Dextran 40
(Comment on this Monograph)id=m23770=Dextran 40=DMonos.pdf) Dextrans [9004-54-0]. DEFINITION Dextran 40 is derived by controlled hydrolysis and fractionation of polysaccharides elaborated by the fermentative action of certain strains of Leuconostoc mesenteroides (NRRL, B.512 F; NCTC, 10817) on a sucrose substrate. It is a glucose polymer in which the linkages between glucose units are almost entirely of the -1:6 type. Its weight average molecular weight is in the 35,00045,000 range. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solutions: Four solutions of Dextran 40 in water, in such a manner that the concentrations are accurately known and approximately evenly distributed in the range of 2%0.5%. Analysis: Using a capillary tube viscosimeter having dimensions such that the flow time of water is NLT 100 s, measure the flow times of water and of the Sample solution at 20. Calculate the viscosity numbers of each of the Sample solutions: Result = {ln[(RD)(t/t0)]}/C RD t t0 = ratio of the density of the individual Sample solution to that of water = flow time for the Sample solution = flow time for the water
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= concentration of Dextran 40 in the Sample solution (g/mL) Acceptance criteria: Plot the viscosity numbers of each of the Sample solutions against their respective concentrations, and draw the straight line of best fit through the points and extrapolate to zero concentration. The value of the intercept is 1823 mL/g. IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 5 ppm CHLORIDE AND SULFATE, Sulfate 221: A 1.5-g portion shows NMT 0.03%, which corresponds to 0.45 mL of 0.020 N sulfuric acid. Organic Impurities PROCEDURE 1: LIMIT OF NITROGENOUS IMPURITIES (where it is labeled as intended for use in the preparation of injectables) Sulfate solution: To 1000 mL of sulfuric acid add 5 g of anhydrous cupric sulfate and 500 g of potassium sulfate. Dissolve by heating, and store at 60. [NOTEIf storage at 60 is not possible, prepare a smaller quantity of Sulfate solution on the day of use, adjusting the proportions accordingly.] Indicator: Dilute a mixture of 20 mL of a 0.1% solution of bromocresol green in alcohol and 4 mL of methyl red TS with water to 100 mL. Analysis Sample: 0.2 g of Dextran 40 Transfer Sample to a micro-Kjeldahl flask. Add 4 mL of Sulfate solution to the Sample. Heat until the solution exhibits a clear green color and the sides of the flask are free from carbonaceous material. Cool, cautiously add 30 mL of water, and transfer the solution to a steam distillation unit. Rinse the Kjeldahl flask with three 5-mL portions of water, adding the washings to the solution. Add 15 mL of 45% sodium hydroxide solution, immediately close the distillation apparatus, and start steam distillation immediately. Receive the distillate in 1 mL of Indicator and sufficient water to cover the end of the condensing tube. Upon completion of the distillation, remove the receiving flask, and rinse the end of the condensing tube with a small quantity of water, adding the rinse to the distillate. Titrate the distillate with 0.010 N hydrochloric acid until the color changes from blue to reddish violet. Perform a blank determination, and make any necessary correction. Acceptance criteria: The corrected volume of 0.010 N hydrochloric acid titrated does not exceed 0.14 mL (0.01%, as N). PROCEDURE 2: LIMIT OF ALCOHOL AND RELATED IMPURITIES Sample solution: Dissolve without heating 5.0 g in 100 mL of water, and distill the solution, collecting the first 45 mL of the distillate. Dilute the distillate with water to 50.0 mL. Standard solution: To 25.0 mL of the Sample solution add 0.5 mL of a 2.5% (w/v) solution of n-propyl alcohol. Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m; packed with S3 Temperature Column: 160 Injection port: 240 Detector: 210 Carrier gas: Nitrogen Flow rate: 25 mL/min [NOTEInjector seals may deteriorate after multiple injections of the Standard solution and Sample solution. Inspect the seals before making a series of injections.] C

Injection size: 1 L Analysis Samples: Standard solution, Sample solution, and a 0.05% (w/v) solution of n-propyl alcohol and water Acceptance criteria: After corrections for any impurities in the n-propyl alcohol solution and water, the total area of peaks from impurities in the Sample solution does not exceed the area of the n-propyl alcohol solution peak. PROCEDURE 3: ANTIGENIC IMPURITIES (where it is labeled as intended for use in the preparation of injectables) Sterile solution: 100 mg/mL in sodium chloride injection Analysis: At intervals of about 48 h, inject three 0.5-mL doses into the peritoneal cavities of each of six guinea pigs. At 14 days after the first intraperitoneal injection, inject 0.20 mL intravenously into each of three of the guinea pigs, and at 21 days treat the other three guinea pigs similarly. Observe the animals for 30 min after each intravenous injection and again 24 h later. Acceptance criteria: The animals exhibit no evidence of anaphylactoid reactions, such as coughing, bristling of hair, or respiratory distress. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +195 to +203 Sample solution: 20 mg/mL, heated, if necessary, on a water bath to dissolve PH 791: 4.57.0, in a solution (1 in 10) LOSS ON DRYING 731: Dry it at 105 for 5 h: it loses NMT 7.0% of its weight. BACTERIAL ENDOTOXINS TEST 85 (where it is labeled as intended for use in the preparation of injectables): When tested in Sodium Chloride Injection (1 in 10), it contains NMT 1.0 USP Endotoxin Unit/mL. COLOR OF SOLUTION: The absorbance of a solution in water (1 in 10), measured in a 4-cm cell determined at 375 nm against a water blank, is NMT 0.20. SAFETY: Inject intravenously 1.0 mL of a sterile solution (1 in 10) of 10% Dextran 40 in saline TS into each of five mice weighing 1820 g. The injection period is 1015 s. If there are no deaths within 72 h, it meets the requirements of the test. If one or more animals die, continue the test using 10 mice weighing 20 0.5 g. If all animals survive for 72 h, the requirements of the test are met. MOLECULAR WEIGHT DISTRIBUTION and WEIGHT and NUMBER AVERAGE MOLECULAR WEIGHTS Mobile phase: 7.1 mg/mL of anhydrous sodium sulfate Calibration solutions: Separately dissolve USP Dextran 4 Calibration RS, USP Dextran 10 Calibration RS, USP Dextran 40 Calibration RS, USP Dextran 70 Calibration RS, and USP Dextran 250 Calibration RS in Mobile phase to obtain solutions each containing 20 mg/mL of the Reference Standard. Marker solution: A solution in Mobile phase containing 3 mg of dextrose and 3 mg of USP Dextran V0 Marker RS/mL System suitability solution: 20 mg/mL of USP Dextran 40 System Suitability RS in Mobile phase Sample solution: 20 mg/mL of Dextran 40 in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: Three 7.5-mm 30-cm columns; packing L38, maintained at a constant temperature Injection size: 50 L System suitability Samples: Calibration solutions, Marker solution, and System suitability solution Suitability requirements Elution profile: Profile shows two peaks, the first due to the V0 marker, the second due to dextrose, Marker solution
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Tailing factor: NMT 1.3, for the dextrose peak, Marker soluton Relative standard deviation of the ratio V0/VT: Determine the void volume, V0, of the system as the inflection point of the ascending part of the first peak. Determine the total volume, VT, of the system as the maximum of the second peak. The relative standard deviation of V0/VT is NMT 1%, Marker solution Molecular weight check: Chromatograph each of the Calibration solutions separately. Divide each profile into at least 60 vertical sections of equal volume increments. (The actual number of sections is represented by the variable a in the equations below.) Record yi, the height above the baseline, corresponding to each value of vi, the volume eluted at that section. For each value of vi, calculate the distribution coefficient, Ki: (vi V0)/(VT V0) Find appropriate values of b1, b2, b3, b4, and b5, using a suitable method, that, when substituted in the equation:
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Similarly, calculate Mw of the low-fraction dextran eluted in and after section m:
in which m is defined by:
and the resulting values of Mi substituted, along with their corresponding values of yi, in the equation:
give values of weight average molecular weight, Mw, within 5% of the labeled values for each of the Calibration solutions and 180 2 for dextrose, Calibration solutions. Average Mw check: Using the System suitability solution, calculate Mw of the total molecular weight distribution using the same method as directed for the Molecular weight check, but inserting the now known values of b1, b2, b3, b4, and b5. It is between 39,000 and 46,000. Similarly, calculate Mw of the high-fraction dextran eluted through section n:
It is between 6000 and 9000, System suitability solution. Analysis Sample: Sample solution Calculate values of the weight average molecular weight, Mw, of the total molecular weight distribution of the highfraction dextran, and of the low-fraction dextran. With the values of b1, b2, b3, b4, and b5, obtained with the Calibration solutions under Chromatographic system, calculate the number average molecular weight, Mn, of the total molecular weight distribution of the Sample solution by substituting the corresponding values of Mi, along with their corresponding values of yi, in the equation:
Acceptance criteria: The weight average molecular weight of the total molecular weight distribution of the high-fraction dextran, and of the low-fraction dextran are 35,00045,000 (NMT 120,000 and NLT 5,000). The number average molecular weight, Mn, is between 16,000 and 30,000. Where Dextran 40 is labeled as intended for use in the preparation of injectables, the ratio Mw/Mn is in the 1.41.9 range. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at 25, excursions permitted between 15 and 30. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Dextran 40 RS USP Dextran 4 Calibration RS USP Dextran 10 Calibration RS USP Dextran 40 Calibration RS USP Dextran 70 Calibration RS USP Dextran 250 Calibration RS USP Dextran Vo Marker RS USP Dextran 40 System Suitability RS USP Endotoxin RS
in which n is defined by the relations:
It is between 111,000 and 135,000.

Gauss-Newton method, modified by Hartley [see D. Hartley Technometrics, 3 (1961)], and the G. Nilsson and K. Nilsson method [see G. Nilsson and K. Nilsson J. Chromat., 101, 137 (1974)] are suitable methods. A curve-fitting program capable of nonlinear regression may be used.
The
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Mr2 = molecular weight of dextrose, 180.16 Acceptance criteria: 4.15.0 g DEXTRAN 40 Sample solution: To 25 mL of volume of Injection, add 1 drop of 5 N ammonium hydroxide. Analysis: Determine the optical rotation (see Optical Rotation 781). Calculate the concentration, in g/100 mL, of dextran 40 in the Injection taken: Result = (1/Av1)[(100a/l) (Av2Cd)] = average value for the specific rotation of dextran 40, 197.5 a = observed optical rotation () l = length of the polarimeter tube (dm) = average value for the specific rotation of Av2 dextrose, 52.75 = concentration of dextrose as determined in the Cd assay for Dextrose (g/100 mL) Acceptance criteria: 9.011.0 g Av1 IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 5 ppm Organic Impurities PROCEDURE: LIMIT OF 5-HYDROXYMETHYLFURFURAL AND RELATED SUBSTANCES Sample solution: 2 mg/mL of C6H12O6 H2O from volume of Injection, in water Spectrometric conditions Cell: 1 cm Analytical wavelength: 284 nm Blank: Water Analysis Samples: Sample solution and Blank Acceptance criteria: NMT 0.25 SPECIFIC TESTS PH 791: 3.07.0 STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.0 USP Endotoxin Unit/mL. COLOR OF SOLUTION: Its absorbance, determined at 375 nm against a water blank, is NMT 0.06. OTHER REQUIREMENTS: It meets the requirements for Injections 1 and for Particulate Matter in Injections 788. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose glass or plastic containers. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, the label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Dextrose RS USP Endotoxin RS
Dextran 40 in Dextrose Injection

(Comment on this Monograph)id=m23800=Dextran 40 in Dextrose Injection=D-Monos.pdf) DEFINITION Dextran 40 in Dextrose Injection is a sterile solution of Dextran 40 and Dextrose in Water for Injection. It contains in each 100 mL NLT 9.0 g and NMT 11.0 g of Dextran 40 and NLT 4.1 g and NMT 5.0 g of dextrose (C6H12O6). It contains no bacteriostatic agents. IDENTIFICATION PROCEDURE Comparison solution: Dextrose (4.5 in 100) Sample solution: 10 mg of dextran 40/mL from volume of Injection, in dextrose solution (4.5 in 100) Analysis: Using a capillary tube viscosimeter having dimensions such that the flow time of water is NLT 100 s, measure the flow times of the Comparison solution and of the Sample solution at 20. Calculate the intrinsic viscosity taken: Result = {ln[(RD)(t/t0)]}/C = ratio of the density of the Sample solution to that of the Comparison solution t = flow time for the Sample solution t0 = flow time for the Comparison solution C = concentration of dextran 40 in the Sample solution (mg/mL) Acceptance criteria: The intrinsic viscosity is 1823 mL/g. ASSAY DEXTROSE Mobile phase: 0.01 N sulfuric acid System suitability solution: 5 mg/mL each of dextrose and xylitol in water Standard solution: 5 mg/mL of dextrose monohydrate of USP Dextrose RS in water Sample solution: 5 mg/mL of dextrose monohydrate from volume of Injection in water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: 7.8-mm 30-cm; packing L17 Temperature: Column and detector (if necessary) are maintained at a constant temperature of 40 Flow rate: 0.6 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 2.5 between the dextrose and xylitol peaks, System suitability solution Relative standard deviation: NMT 1.5% for dextrose, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of C6H12O6 H2O in the volume of Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS CS CU Mr1 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Dextrose RS in the Standard solution (mg/mL) = nominal concentration of dextrose monohydrate in the Sample solution (mg/mL) = molecular weight of dextrose monohydrate, 198.17 RD
Dextran 40 in Sodium Chloride Injection

(Comment on this Monograph)id=m23830=Dextran 40 in Sodium Chloride Injection=D-Monos.pdf) DEFINITION Dextran 40 in Sodium Chloride Injection is a sterile solution of Dextran 40 and Sodium Chloride in Water for Injection. It contains in each 100 mL NLT 9.0 g and NMT 11.0 g of
66
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label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Endotoxin RS
dextran 40; and NLT 0.81 g and NMT 0.99 g of sodium chloride. It contains no bacteriostatic agents. IDENTIFICATION A. PROCEDURE Comparison solution: Sodium chloride solution (0.9 in 100) Sample solution: 10 mg of dextran 40/mL from a volume of Injection, in sodium chloride solution (0.9 in 100) Analysis: Using a capillary tube viscosimeter with a flow time for water of NLT 100 s, measure the flow times of the Comparison solution and of the Sample solution at 20. Calculate the intrinsic viscosity: Result = {ln[(RD)(t/t0)]}/C = ratio of the density of the Sample solution to that of the Comparison solution t = flow time for the Sample solution t0 = flow time for the Comparison solution C = concentration of dextran 40 in the Sample solution (mg/mL) Acceptance criteria: The intrinsic viscosity is 1823 mL/g. RD ASSAY SODIUM CHLORIDE Sample solution: Equivalent to 90 mg of chloride from a volume of Injection into a porcelain casserole, and add 100 mL of water and 1 mL of dichlorofluorescein TS. Analysis: Titrate with 0.1 N silver nitrate VS until the silver chloride flocculates and the mixture acquires a faint pink color. Each mL of 0.1 N silver nitrate is equivalent to 5.844 mg of sodium chloride. Acceptance criteria: It contains NLT 0.81 g and NMT 0.99 g of sodium chloride/100 mL. DEXTRAN 40 Sample solution: To 25 mL of Injection, add 1 drop of 5 N ammonium hydroxide. Analysis: Determine the optical rotation (see Optical Rotation 781). Calculate the concentration, in g/100 mL, of dextran 40 in the Injection taken: Result = 100(a/l )(1/Av) = observed optical rotation (degrees) = length of the polarimeter tube (dm) = average value for the specific rotation of dextran 40, 197.5 Acceptance criteria: It contains NLT 9.0 g and NMT 11.0 g of dextran 40/100 mL. IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: a l Av
Dextran 70
(Comment on this Monograph)id=m23855=Dextran 70=DMonos.pdf) Dextrans. DEFINITION Dextran 70 is derived by controlled hydrolysis and fractionation of polysaccharides elaborated by the fermentative action of certain appropriate strains of Leuconostoc mesenteroides (NRRL, B-512F; NCTC, 10817) on a sucrose substrate. It is a glucose polymer in which the linkages between glucose units are almost entirely of the -1:6 type. Its weight average molecular weight is in the 63,00077,000 range. IDENTIFICATION A. INFRARED ABSORPTION 197K B. PROCEDURE Sample solutions: Prepare four solutions of Dextran 70 in water in such a manner that the concentrations are accurately known and approximately evenly distributed in the range of 2%0.5%. Analysis: Using a capillary tube viscosimeter having dimensions such that the flow time of water is NLT 100 s, measure the flow times of water and of the Sample solution at 20. Calculate the viscosity numbers of each of the Sample solutions: Result = {ln[(RD)(t/t0)]}/C = ratio of the density of the individual Sample solution to that of water t = flow time for the Sample solution = flow time for the water t0 C = concentration of Dextran 70 in the Sample solution (g/mL) Plot the viscosity numbers of each of the Sample solutions against their respective concentrations, and draw the straight line of best fit through the points, and extrapolate to zero concentration. Acceptance criteria: The value of the intercept is between 24 and 29 mL/g. RD IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 5 ppm CHLORIDE AND SULFATE, Sulfate 221: A 1.5-g portion shows no more sulfate than corresponds to 0.45 mL of 0.020 N sulfuric acid (0.03%). Organic Impurities PROCEDURE 1: LIMIT OF NITROGENOUS IMPURITIES (where it is labeled as intended for use in the preparation of injectables) Sulfate solution: To 1000 mL of sulfuric acid, add 5 g of anhydrous cupric sulfate and 500 g of potassium sulfate. Dissolve by heating, and store at 60. [NOTEIf storage at 60 is not possible, prepare a smaller quantity of Sulfate solution on the day of use, adjusting the proportions accordingly.] Indicator: Dilute a mixture of 20 mL of a 0.1% solution of bromocresol green in alcohol and 4 mL of methyl red TS with water to 100 mL.
NMT 5 ppm
SPECIFIC TESTS PH 791: 3.57.0 STERILITY 71: It meets the requirements when tested as directed for Test for Sterility of the Product to be Examined, Membrane Filtration. OTHER REQUIREMENTS: It meets the requirements for Injections 1 and for Particulate Matter in Injections 788. BACTERIAL ENDOTOXINS TEST 85: It contains NMT 1.0 USP Endotoxin Unit/mL. COLOR OF SOLUTION: Its absorbance, determined at 375 nm against a water blank, is NMT 0.05. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose glass or plastic containers. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, the
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Analysis Sample: 0.2 g of Dextran 70 Transfer Sample to a micro-Kjeldahl flask. Add 4 mL of Sulfate solution. Heat until the solution exhibits a clear green color and the sides of the flask are free from carbonaceous material. Cool, and transfer the solution to a steam distillation unit. Rinse the Kjeldahl flask three times with 5 mL of water, adding the washings to the solution. Add 15 mL of 45% sodium hydroxide solution, immediately close the distillation apparatus, and commence steam distillation without delay. Receive the distillate in 1 mL of Indicator in a 100-mL flask, keeping the end of the condensing tube below the liquid surface for 5 min and above the liquid surface for 1 min. Upon completing the distillation, remove the receiving flask, and rinse the end of the condensing tube with a small quantity of water, adding the rinse to the distillate. Titrate the distillate with 0.010 N hydrochloric acid until the color changes from blue to reddish violet. Perform a blank determination, and make any necessary correction. Acceptance criteria: The corrected volume of 0.010 N hydrochloric acid titrated does not exceed 0.14 mL (0.01%, as N). PROCEDURE 2: LIMIT OF ALCOHOL AND RELATED IMPURITIES Sample solution: Dissolve without heating 5.0 g in 100 mL of water, and distill the solution, collecting the first 45 mL of the distillate. Dilute the distillate with water to 50.0 mL. Standard solution: To 25.0 mL of the Sample solution, add 0.5 mL of a 2.5% (w/v) solution of n-propyl alcohol Chromatographic system (See Chromatography 621, System Suitability.) Mode: GC Detector: Flame ionization Column: 2-mm 1.8-m; packed with support S3 Temperature Column: 160 Injection port: 240 Detector: 210 Carrier gas: Nitrogen Flow rate: 25 mL/min [NOTEInjector seals may deteriorate after multiple injections of the Standard solution and Sample solution. Inspect the seals before making a series of injections.] Injection size: 1 L Analysis Samples: Standard solution, Sample solution, and a 0.05% (w/v) solution of n-propyl alcohol and water Acceptance criteria: After corrections for any impurities in the n-propyl alcohol solution and water, the total area of peaks from impurities in the Sample solution does not exceed the area of the n-propyl alcohol solution peak. SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +195 to +203 Sample solution: 20 mg/mL, heated, if necessary, on a water bath to dissolve PH 791: 4.57.0, in a solution (6 in 100) LOSS ON DRYING 731: Dry it at 105 for 5 h: it loses NMT 7.0% of its weight. BACTERIAL ENDOTOXINS TEST 85 (where it is labeled as intended for use in the preparation of injectables): When tested in Sodium Chloride Injection (0.6 in 10), it contains NMT 0.5 USP Endotoxin Unit/mL. COLOR OF SOLUTION: The absorbance of a solution in water (6 in 100), measured in a 4-cm cell determined at 375 nm against a water blank, is NMT 0.15. SAFETY: Inject intravenously 1.0 mL of a sterile solution of 6% Dextran 70 in saline TS into each of five mice weighing

1820 g. The injection period is NLT 10 s and NMT 15 s. If there are no deaths within 72 h, it meets the requirements of the test. If 1 or more animals die, continue the test using 10 mice weighing 20 0.5 g. If all animals survive for 72 h, the requirements of the test are met. ANTIGENIC IMPURITIES (where it is labeled as intended for use in the preparation of injectables): Prepare a sterile solution containing 60 mg/mL of Dextran 70 in Sodium Chloride Injection. At intervals of about 48 h, inject three 0.5-mL doses into the peritoneal cavities of each of 6 guinea pigs. At 14 days after the first intraperitoneal injection, inject 0.20 mL intravenously into each of 3 of the guinea pigs, and at 21 days treat the other 3 guinea pigs similarly. Observe the animals for 30 min after each intravenous injection and again 24 h later. The animals exhibit no evidence of anaphylactoid reactions, such as coughing, bristling of hair, or respiratory distress. MOLECULAR WEIGHT DISTRIBUTION AND WEIGHT AND NUMBER AVERAGE MOLECULAR WEIGHTS Mobile phase: 7.1 g/L of anhydrous sodium sulfate in water Calibration solutions: 20 mg/mL each of USP Dextran 4 Calibration RS, USP Dextran 10 Calibration RS, USP Dextran 40 Calibration RS, USP Dextran 70 Calibration RS, and USP Dextran 250 Calibration RS in Mobile phase Marker solution: 3 mg/mL each of dextrose and USP Dextran V0 Marker RS in Mobile phase System suitability solution: 20 mg/mL of USP Dextran 70 System Suitability RS in Mobile phase Sample solution: 20 mg/mL of Dextran 70 in Mobile phase Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: Three 7.5-mm 30-cm columns; packing L38 Temperature: Constant temperature Injection size: 50 L System suitability Samples: Calibration solutions, Marker solution, and System suitability solution Suitability requirements Elution profile: Profile shows two peaks: the first due to the V0 marker, and the second due to dextrose, Marker solution Tailing factor: NMT 1.3 for the dextrose peak, Marker solution Relative standard deviationof the ratio V0/VT: Determine the void volume, V0, of the system as the inflection point of the ascending part of the first peak. Determine the total volume, VT, of the system as the maximum of the second peak. The relative standard deviation of V0/VT is NMT 1.0%, Marker solution Molecular weight check: Chromatograph each of the Calibration solutions separately. Divide each profile into at least 60 vertical sections of equal volume increments. (The actual number of sections is represented by the variable a in the equations below.) Record yi, the height above the baseline, corresponding to each value of vi, the volume eluted at that section. For each value of vi, calculate the distribution coefficient, Ki: Result = (vi V0)/(VT V0) Find appropriate values of b1, b2, b3, b4, and b5 using a suitable method that, when substituted in the equation:
68

and the resulting values of Mi substituted, along with their corresponding values of yi, in the equation:
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solution by substituting the corresponding values of Mi, along with their corresponding values of yi:
give values of weight average molecular weight, Mw, within 5% of the labeled values for each of the Calibration solutions and 180 2 for dextrose. Average Mw check: Using the System suitability solution, calculate Mw of the total molecular weight distribution using the same method as directed for the Molecular weight check, but inserting the now known values of b1, b2, b3, b4, and b5. It is between 65,000 and 74,000. Similarly, calculate Mw of the high-fraction dextran eluted through section n:
Acceptance criteria: the weight average molecular weight, Mw, of the total molecular weight distribution of the highfraction dextran, and of the low-fraction dextran are between 63,000 and 77,000 (NMT 195,000 and NLT 13,000, respectively). The number average molecular weight, Mn, is between 34,000 and 48,000. Where Dextran 70 is labeled as intended for use in the preparation of injectables, the ratio Mw/Mn is in the 1.41.9 range. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at 25; excursions are permitted between 15 and 30. LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms. USP REFERENCE STANDARDS 11 USP Dextran 70 RS USP Dextran 4 Calibration RS USP Dextran 10 Calibration RS USP Dextran 40 Calibration RS USP Dextran 70 Calibration RS USP Dextran 250 Calibration RS USP Dextran V0 Marker RS USP Dextran 70 System Suitability RS USP Endotoxin RS
= defined by the relations:
It is between 180,000 and 240,000. Similarly, calculate Mw of the low-fraction dextran eluted in and after section m:
Dextran 70 in Dextrose Injection

(Comment on this Monograph)id=m23860=Dextran 70 in Dextrose Injection=D-Monos.pdf) DEFINITION Dextran 70 in Dextrose Injection is a sterile solution of Dextran 70 and Dextrose in Water for Injection. It contains in each 100 mL NLT 5.4 g and NMT 6.6 g of dextran 70 and NLT 4.1 g and NMT 5.0 g of dextrose (C6H12O6). It contains no bacteriostatic agents. IDENTIFICATION A. PROCEDURE Comparison solution: Dextrose (4.5 in 100) Sample solution: 10 mg of dextran 70/mL from volume of Injection, in dextrose solution (4.5 in 100) Analysis: Using a capillary tube viscosimeter having dimensions such that the flow time of water is NLT 100 s, measure the flow times of the Comparison solution and of the Sample solution at 20. Calculate the intrinsic viscosity taken: Result = {ln[(RD)(t/t0)]}/C RD t t0 C = ratio of the density of the Sample solution to that of the Comparison solution = flow time for the Sample solution = flow time for the Comparison solution = concentration of dextran 70 in the Sample solution (mg/mL)
in which m is defined by:
It is between 7,000 and 11,000. Analysis Sample: Sample solution Calculate values of the weight average molecular weight, Mw, of the total molecular weight distribution of the highfraction dextran, and of the low-fraction dextran. With the values of b1, b2, b3, b4, and b5 obtained with the Calibration solutions under Chromatographic system, calculate the number average molecular weight, Mn, of the total molecular weight distribution of the Sample
USP 32
Acceptance criteria: The intrinsic viscosity is 2429 mL/g.

Spectrometric conditions Cell: 1 cm Analytical wavelength: 284 nm Blank: Water Analysis Samples: Sample solution and Blank Acceptance criteria: NMT 0.25 SPECIFIC TESTS PH 791: 3.57.0 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.5 USP Endotoxin Unit/mL. STERILITY TESTS 71: It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration. COLOR OF SOLUTION: Its absorbance, determined at 375 nm against a water blank, is NMT 0.05. OTHER REQUIREMENTS: It meets the requirements for Injections 1 and for Particulate Matter in Injections 788. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose glass or plastic containers. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, the label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Dextrose RS USP Endotoxin RS
ASSAY DEXTROSE Mobile phase: 0.01 N sulfuric acid System suitability solution: 5 mg/mL each of dextrose and xylitol in water Standard solution: 5 mg/mL of dextrose monohydrate of USP Dextrose RS in water Sample solution: 10 mL of volume of Injection in 25 mL of water Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: Refractive index Column: 7.8-mm 30-cm; packing L17 Temperature: Column and detector, if necessary, are maintained at a constant temperature of about 40. Flow rate: 0.6 mL/min Injection size: 50 L System suitability Samples: System suitability solution and Standard solution Suitability requirements Resolution: NLT 2.5 between the dextrose and xylitol peaks, System suitability solution Relative standard deviation: NMT 1.5% for dextrose, Standard solution Analysis Samples: Standard solution and Sample solution Calculate the percentage of dextrose monohydrate (C6H12O6 H2O) in the volume of Injection taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 = peak area from the Sample solution = peak area from the Standard solution = concentration of USP Dextrose RS in the Standard solution (mg/mL) = nominal concentration of dextrose monohydrate CU in the Sample solution (mg/mL) Mr1 = molecular weight of dextrose monohydrate, 198.17 Mr2 = molecular weight of dextrose, 180.16 Acceptance criteria: 4.15.0 g of C6H12O6/100 mL DEXTRAN 70 Sample solution: To 25 mL of volume of Injection add 1 drop of 5 N ammonium hydroxide. Analysis: Determine the optical rotation (see Optical Rotation 781). Calculate the concentration, in g/100 mL, of dextran 70 in the Injection taken: Result = (1/AV1)[(100a/l) (AV2C)] = average value for the specific rotation of dextran 70, 197.5 a = observed optical rotation (degrees) l = length of the polarimeter tube (dm) = average value for the specific rotation of AV2 dextrose, 52.75 C = concentration of dextrose as determined in the Assay for Dextrose (g/100 mL) Acceptance criteria: 5.46.6 g of dextran 70/100 mL AV1 IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: NMT 5 ppm Organic Impurities PROCEDURE: LIMIT OF 5-HYDROXYMETHYLFURFURAL AND RELATED SUBSTANCES Sample solution: 2 mg/mL of C6H12O6 H2O from volume of Injection, in water rU rS CS
Dextran 70 in Sodium Chloride Injection

(Comment on this Monograph)id=m23865=Dextran 70 in Sodium Chloride Injection=D-Monos.pdf) DEFINITION Dextran 70 in Sodium Chloride Injection is a sterile solution of Dextran 70 and Sodium Chloride in Water for Injection. It contains, in each 100 mL, NLT 5.4 g and NMT 6.6 g of dextran 70, and NLT 0.81 g and NMT 0.99 g of sodium chloride. It contains no bacteriostatic agents. IDENTIFICATION PROCEDURE Sample solution: Quantitatively dilute a portion of Injection with sodium chloride solution (0.9 in 100) to a concentration of 10 mg/mL of dextran 70. Analysis: Using a capillary tube viscosimeter having dimensions such that the flow time of water is NLT 100 s, measure the times of the diluted Injection and sodium chloride solution (0.9 in 100) at 20. Calculate the intrinsic viscosity: Result = {ln[(RD) (t/t0)]}/C = ratio of the density of the diluted Injection to that of the sodium chloride solution t = flow time for the diluted Injection = flow time for the sodium chloride solution t0 C = concentration of dextran 70 in the diluted Injection (g/mL) Acceptance criteria: 2429 mL/g RD ASSAY SODIUM CHLORIDE Sample solution: Pipet a volume of Injection, equivalent to 90 mg of chloride, into a porcelain casserole, and add 100 mL of water and 1 mL of dichlorofluorescein TS.
70
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of 1 volume of benzoyl chloride and 2 volumes of anhydrous ethyl ether, insert the stopper, and shake for 3 min. Filter the precipitate, wash it with about 10 mL of cold water, and recrystallize it from diluted alcohol: the crystals of the benzoyl derivative of dextroamphetamine so obtained, after being dried at 105 for 3 h, melt between 154 and 160. B. IDENTIFICATION TESTSGENERAL, Sulfate 191: A solution (100 mg/mL) meets the requirements. ASSAY PROCEDURE Sample solution: 10 mg/mL of Dextroamphetamine Sulfate in glacial acetic acid Analysis: Titrate with 0.1 N perchloric acid VS. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 36.85 mg of (C9H13N)2 H2SO4. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE Solution A: 2.16 mg/mL of sodium 1-octanesulfonate in water. Add 1.0 mL of triethylamine/L of octanesulfonate solution. Adjust with phosphoric acid to a pH of 2.5. Mobile phase: Acetonitrile, methanol, and Solution A (37:19:144) Diluent: Dilute 3.12 mL of phosphoric acid with water to 1000 mL Standard stock solution: 0.3 mg/mL of USP Dextroamphetamine Sulfate RS in Diluent. Standard solution: 0.003 mg/mL from Standard stock solution in Diluent Sample solution: 0.3 mg/mL of Dextroamphetamine Sulfate in Diluent [NOTESonicate for 5 min, and then dilute with Diluent to volume.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 215 nm Column: 4.6-mm 15-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 50 L System suitability Sample: Standard stock solution and Sample solution Suitability requirements Resolution: NLT 1.5 between dextroamphetamine and any adjacent peak Tailing factor: NMT 2.0 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of each impurity in the portion of Dextroamphetamine Sulfate taken: Result = (ri/rS) (CS/W) 10,000 ri
Analysis: Titrate with 0.1 N silver nitrate VS until the silver chloride flocculates and the mixture acquires a faint pink color. Each mL of 0.1 N silver nitrate is equivalent to 5.844 mg of sodium chloride. Acceptance criteria: 0.810.99 g DEXTRAN 70 Sample solution: To 25 mL of Injection add 1 drop of 5 N ammonium hydroxide. Analysis: Determine the optical rotation (see Optical Rotation 781) in a suitable polarimeter. Calculate the concentration, in g/100 mL, of dextran 70 in the portion of Injection taken: Result = 100 (a/L) (1/AV) = observed optical rotation in degrees = length of the polarimeter tube (dm) = average value for the specific rotation of dextran 70, 197.5 Acceptance criteria: 5.46.6g/100 mL IMPURITIES Inorganic Impurities HEAVY METALS, Method II 231: a L AV
NMT 5 ppm
SPECIFIC TESTS PH 791: 4.07.0 BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.5 USP Endotoxin Unit/mL. STERILITY TESTS 71: It meets the requirements when tested as directed under Test for Sterility of the Product to Be Examined, Membrane Filtration. COLOR OF SOLUTION: Its absorbance, determined at 375 nm against a water blank, is NMT 0.04. OTHER REQUIREMENTS: It meets the requirements for Injections 1 and Particulate Matter in Injections 788. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in a single-dose glass or in plastic containers. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, the label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Dextroamphetamine Sulfate
(Comment on this Monograph)id=m24040=Dextroamphetamine Sulfate=DMonos.pdf)
(C9H13N)2 H2SO4 Benzeneethanamine, -methyl-, (S)-, sulfate (2:1); (+)--Methylphenethylamine sulfate (2:1) [51-63-8].
368.49
rS CS W
DEFINITION Dextroamphetamine Sulfate, the dextrorotatory isomer of amphetamine sulfate, contains NLT 98.0% and NMT 101.0% of (C9H13N)2 H2SO4, calculated on the dried basis. IDENTIFICATION A. Dissolve 100 mg in 5 mL of water, add 5 mL of 1 N sodium hydroxide, cool to 1015, add 1 mL of a mixture
= peak response for each impurity of the Sample solution = peak response of the Standard solution = concentration of USP Dextroamphetamine Sulfate RS in the Standard solution (mg/mL) = weight of Dextroamphetamine Sulfate taken to prepare the Sample solution (mg)
USP 32
Acceptance criteria Individual impurities: NMT 0.1% Total impurities: NMT 0.5% SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: +20 to +23.5 Sample solution: 40 mg/mL PH 791: 5.06.0, in a solution (1 in 20) LOSS ON DRYING 731: Dry it at 105 for 2 h: it loses NMT 1.0% of its weight. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dextroamphetamine Sulfate RS
Official Monographs / Dextroamphetamine 71

Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C9H13N)2 H2SO4 in the portion of Capsules taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dextroamphetamine Sulfate RS in the Standard solution (mg/mL) = nominal concentration of dextroamphetamine CU sulfate in the Sample solution (mg/mL) Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 500 mL Apparatus 1: 100 rpm Time: 45 min Analysis: Determine the amount of (C9H13N)2 H2SO4 dissolved, employing the analysis set forth in the Assay, making any necessary modifications. Tolerances: NLT 75% (Q) of the labeled amount of (C9H13N)2 H2SO4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dextroamphetamine Sulfate RS rU rS CS
Dextroamphetamine Sulfate Capsules

(Comment on this Monograph)id=m24043=Dextroamphetamine Sulfate Capsules=D-Monos.pdf) DEFINITION Dextroamphetamine Sulfate Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of (C9H13N)2 H2SO4. IDENTIFICATION A. PROCEDURE Sample solution: Mix an amount of the Capsule contents, equivalent to 50 mg of dextroamphetamine sulfate, with 10 mL of water for 30 min, and filter into a small flask. Analysis: Cool the filtrate to 15, add 3 mL of 1 N sodium hydroxide, then add 1 mL of a mixture of 1 volume of benzoyl chloride and 2 volumes of anhydrous ethyl ether, and shake for 2 min. Filter the precipitate, wash with 15 mL of cold water, and recrystallize twice from diluted alcohol. Acceptance criteria: The benzoyl derivative of dextroamphetamine so obtained, after being dried at 105 for 1 h, melts between 154 and 160. B. The retention time of the major peak of the Sample solution is the same as that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 1.1 g of sodium 1-heptanesulfonate in 525 mL of water. Add 25 mL of dilute glacial acetic acid (14 in 100) and 450 mL of methanol. Adjust dropwise, if necessary, with glacial acetic acid to a pH of 3.3 0.1. The volume of methanol may be adjusted so that the retention time for dextroamphetamine is about 5 min. Pass through a 0.5-m membrane filter. Standard solution: 0.3 mg/mL of USP Dextroamphetamine Sulfate RS in 0.12 N phosphoric acid Sample solution: Remove, as completely as possible, the contents of NLT 20 Capsules. Transfer a portion of the mixed powder, equivalent to 15 mg dextroamphetamine sulfate, to a 50-mL volumetric flask. Add 40 mL of 0.12 N phosphoric acid, sonicate for 15 min, and dilute with 0.12 N phosphoric acid to volume. Pass through a 0.5-m membrane filter, discarding the first 20 mL of the filtrate. Chromatographic system (See Chromatography 621, System Suitability.)
Dextroamphetamine Sulfate Tablets

(Comment on this Monograph)id=m24060=Dextroamphetamine Sulfate Tablets=DMonos.pdf) DEFINITION Dextroamphetamine Sulfate Tablets contain NLT 93.0% and NMT 107.0% of the labeled amount of (C9H13N)2 H2SO4. IDENTIFICATION A. Transfer a portion of finely ground Tablets, equivalent to 50 mg of dextroamphetamine sulfate, to a suitable centrifuge tube. Add 25 mL of water, shake vigorously, and centrifuge until clear. Decant the clear solution into a 250mL separator, add 5 mL of 2.5 N sodium hydroxide, and extract with 60 mL of ether. Wash the ether extract with two 5-mL portions of 0.25 N sodium hydroxide, and discard the washings. Filter the ether extract through a pledget of cotton, previously saturated with ether, into a 100-mL beaker, and evaporate on a steam bath in a current of air to 1 mL. Dissolve the residue in 3 mL of alcohol, and transfer
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100) and 400 mL of methanol. Adjust by the dropwise addition of glacial acetic acid to a pH of 3.3 0.1, if necessary, filter, and degas the solution. Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 210 nm Column: 3.9-mm 30-cm; packing L1 Column temperature: 40 Flow rate: 1.5 mL/min Injection size: 100 L System suitability Sample: Standard solution Suitability requirements Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the quantity of (C9H13N)2 H2SO4 dissolved. Tolerances: NLT 75% (Q) of the labeled amount of (C9H13N)2 H2SO4 is dissolved. UNIFORMITY OF DOSAGE UNITS 905: Meet the requirements SPECIFIC TESTS ISOMERIC PURITY: Pack a pledget of fine glass wool in the base of a 200- 25-mm chromatographic tube, with the aid of a tamping rod. Add 5 g of chromatographic siliceous earth, and tamp firmly to compress the material to a uniform mass. Finely powder a number of Tablets, equivalent to 300 mg of dextroamphetamine sulfate, mix the powder in a mortar with 5 g of chromatographic siliceous earth, add 1 mL of methanol and 0.5 mL of ammonium hydroxide, and triturate to a uniform mixture. Transfer the mixture without delay to the chromatographic tube, and tamp as before. Wipe the mortar and pestle with a small amount of glass wool, and insert it into the tube on top of the column. Arrange a 125-mL separator containing 35 mL of 0.1 N sulfuric acid to receive the effluent. Pass 60 mL of chloroform through the column. Shake the separator vigorously for 1 min, allow the layers to separate, and discard the chloroform. Add to the aqueous phase in the separator 2.5 g of sodium bicarbonate, preventing it from coming in contact with the mouth of the separator, swirl until most of the bicarbonate has dissolved. By means of a 1-mL syringe, rapidly inject 1.0 mL of acetic anhydride directly into the contents of the separator. Immediately insert the stopper in the separator, and shake vigorously until the evolution of carbon dioxide has ceased, releasing the pressure as necessary through the stopcock. Allow to stand for 5 min, and extract the solution with 50 mL of chloroform, shaking vigorously for 1 min. Filter the chloroform extract through a pledget of filter cotton into a 100-mL beaker, rinse the cotton with a small amount of chloroform, and evaporate on a steam bath in a current of air or nitrogen to dryness. Heat and triturate the residue until the odor of chloroform is no longer perceptible. Allow the residue to cool, inducing it to crystallize. Reduce the crystals to a fine powder, heat at 80 for 30 min, and cool: the specific rotation of the acetylamphetamine so obtained, determined in a solution in chloroform containing 20 mg/mL, a 200-mm semimicro polarimeter tube being used, is between 37.5 and 44.0 ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. USP REFERENCE STANDARDS 11 USP Dextroamphetamine Sulfate RS
to a glass-stoppered, 125-mL conical flask containing 25 mL of water. Rinse the beaker with 3 mL of alcohol, and transfer to the flask. Cool to 15, add 3 mL of 1 N sodium hydroxide, then add 1 mL of a mixture of 1 volume of benzoyl chloride and 2 volumes of anhydrous ethyl ether, and shake for 2 min. Filter the precipitate, wash with 15 mL of cold water, and recrystallize twice from diluted alcohol: the benzoyl derivative of dextroamphetamine so obtained, after being dried at 105 for 1 h, melts between 154 and 160. B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. ASSAY PROCEDURE Mobile phase: 1.1 g of sodium 1-heptanesulfonate in 525 mL of water. Add 25 mL of dilute glacial acetic acid (14 in 100) and 450 mL of methanol. Adjust dropwise, if necessary, with glacial acetic acid to a pH of 3.3 0.1. Pass through a 0.5-m membrane filter. Standard solution: 0.3 mg/mL of USP Dextroamphetamine Sulfate RS in 0.12 N phosphoric acid Sample solution: Transfer an equivalent to 15 mg of dextroamphetamine sulfate, from finely powdered Tablets (NLT 20), to a 50-mL volumetric flask. Add 40 mL of 0.12 N phosphoric acid, and sonicate for 15 min. Dilute with 0.12 N phosphoric acid to volume. Pass through a 0.5-m membrane filter, discarding the first 20 mL of the filtrate. Chromatographic system (See Chromatography 621 System Suitability.) Mode: LC Detector: UV 254 nm Column: 3.9-mm 30-cm; packing L1 Flow rate: 2 mL/min Injection size: 50 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 3 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of (C9H13N)2 H2SO4 in the portion of Tablets taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dextroamphetamine Sulfate RS in the Standard solution (mg/mL) = concentration of the Sample solution (mg/mL) CU Acceptance criteria: 93.0%107.0% rU rS CS PERFORMANCE TESTS DISSOLUTION, Procedure for a Pooled Sample 711 Medium: Water; 500 mL Apparatus 1: 100 rpm Time: 45 min Standard solution: USP Dextroamphetamine Sulfate RS in Medium. Sample solution: Sample per Dissolution 711. Dilute with Medium to a concentration that is similar to the Standard solution. Determine the amount of (C9H13N)2 H2SO4 dissolved by employing the following method. Mobile phase: 1.1 g of sodium 1-heptanesulfonate in 575 mL of water. Add 25 mL of dilute glacial acetic acid (14 in
USP 32
Official Monographs / Dextromethorphan 73

Acceptance criteria: NMT 0.001% of N,N-dimethylaniline
Dextromethorphan
(Comment on this Monograph)id=m24090=Dextromethorphan=D-Monos.pdf)
SPECIFIC TESTS MELTING RANGE, Class I 741: 109.5112.5 OPTICAL ROTATION, Specific Rotation 781S Sample solution: 100 mg/mL, in chloroform. Its specific rotation does not differ from that of the similarly prepared solution of USP Dextromethorphan RS by more than 1.0%. WATER DETERMINATION, Method Ia 921: NMT 0.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dextromethorphan RS
271.4 C18H25NO Morphinan, 3-methoxy-17-methyl-, (9,13,14)-; 3-Methoxy-17-methyl-9,13,14-morphinan [125-71-3]. DEFINITION Dextromethorphan contains NLT 98.0% and NMT 101.0% of C18H25NO, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 278 nm Sample solution: 100 g/mL in dilute hydrochloric acid (1 in 120) Absorptivities: Calculated on the anhydrous basis, do not differ by more than 3.0%. ASSAY PROCEDURE Sample solution: Dissolve 700 mg of Dextromethorphan in 60 mL of glacial acetic acid. [NOTEWarm slightly, if necessary, to dissolve.] Analysis: Titrate with 0.1 N perchloric acid VS to a bluegreen endpoint, using 2 drops of crystal violet TS as an indicator. Perform a blank determination (see Titrimetry 541). Each mL of 0.1 N perchloric acid is equivalent to 27.14 mg of C18H25NO. Acceptance criteria: 98.0%101.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS, Method II 231: NMT 20 ppm Organic Impurities PROCEDURE 1: LIMIT OF PHENOLIC COMPOUNDS Analysis: Dissolve 10 mg in 2 mL of 3 N hydrochloric acid, add 2 drops of ferric chloride TS, and mix; add 2 drops of potassium ferricyanide TS, and observe after 2 min. Acceptance criteria: No blue-green color develops. PROCEDURE 2: LIMIT OF N,N-DIMETHYLANILINE Standard solution: Transfer 50 mg of N,N-dimethylaniline to a 100-mL volumetric flask, add 70.0 mL of water, insert the stopper tightly, shake for 20 min using a mechanical wrist-action shaker or equivalent, and dilute with water to volume. Transfer 1.0 mL to a 100-mL volumetric flask, and dilute with water to volume. Transfer 1.0 mL of the resulting solution to a 25-mL volumetric flask, and add 19 mL of water. Sample solution: Transfer 500 mg of Dextromethorphan to a 25-mL volumetric flask, add 19 mL of water and 1 mL of 3 N hydrochloric acid, dissolve by warming on a steam bath, and cool. Analysis: Add 2 mL of 1 N acetic acid and 1 mL of sodium nitrite solution (1 in 100) to the Sample solution and dilute with water to volume. This solution shows no more color than the straw yellow to greenish yellow color of the solution of the Standard solution similarly treated.
Dextromethorphan Hydrobromide
(Comment on this Monograph)id=m24120=Dextromethorphan Hydrobromide=D-Monos.pdf) 370.32 C18H25NO HBr H2O Morphinan, 3-methoxy-17-methyl-, (9,13,14)-, hydrobromide, monohydrate; 3-Methoxy-17-methyl-9,13,14-morphinan hydrobromide monohydrate [6700-34-1]. Anhydrous [125-69-9]. 352.32 DEFINITION Dextromethorphan Hydrobromide contains NLT 98.0% and NMT 102.0% of C18H25NO HBr, calculated on the anhydrous basis. IDENTIFICATION A. INFRARED ABSORPTION 197K Sample: Dry in a vacuum over phosphorus pentoxide for 4 h. B. ULTRAVIOLET ABSORPTION 197U Analytical wavelength: 278 nm Sample solution: 100 g/mL in 0.1 N hydrochloric acid Absorptivities: Calculated on the anhydrous basis, do not differ by more than 3.0%. C. To 5 mL of 5 mg/mL solution, add 5 drops of 2 N nitric acid and 2 mL of silver nitrate TS: a yellowish white precipitate is formed. ASSAY PROCEDURE Mobile phase: 0.007 M docusate sodium and 0.007 M ammonium nitrate in a mixture of acetonitrile and water (7:3), and adjust the solution with glacial acetic acid to a pH of 3.4. [NOTEDissolve the docusate sodium in the acetonitrile and water mixture before adding the ammonium nitrate.] Standard solution: 1 mg/mL of USP Dextromethorphan Hydrobromide RS in water. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution: 1 mg/mL of Dextromethorphan Hydrobromide in water. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Chromatographic system (See Chromatography 621, System Suitability.)
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Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr in the portion of Dextromethorphan Hydrobromide taken: Result = (rU/rS) (CS/CU) 100 = peak response from the Sample solution = peak response from the Standard solution = concentration of USP Dextromethorphan Hydrobromide RS in the Standard solution (mg/mL) CU = concentration of dextromethorphan hydrobromide in the Sample solution (mg/mL) Acceptance criteria: 98.0%102.0% rU rS CS IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% Organic Impurities PROCEDURE 1: LIMIT OF PHENOLIC COMPOUNDS: To 5 mg add 1 drop of 3 N hydrochloric acid, 1 mL of water, and 2 drops of ferric chloride TS. Add 2 drops of potassium ferricyanide TS, and observe after 2 min: no blue-green color develops. PROCEDURE 2: LIMIT OF N,N-DIMETHYLANILINE Standard solution: Transfer 50 mg of N,N-dimethylaniline to a 100-mL volumetric flask, add 70.0 mL of water, insert the stopper tightly, shake for 20 min using a mechanical wrist-action shaker or equivalent, and dilute with water to volume. Transfer 1.0 mL to a 100-mL volumetric flask, and dilute with water to volume. Transfer 1.0 mL of the resulting solution to a 25-mL volumetric flask, and add 19 mL of water. Sample solution: Transfer 500 mg of Dextromethorphan Hydrobromide to a 25-mL volumetric flask, add 19 mL of water and 1 mL of 3 N hydrochloric acid, dissolve by warming on a steam bath, and cool. Analysis: Add 2 mL of 1 N acetic acid and 1 mL of sodium nitrite solution (1 in 100) to the Sample solution, and dilute with water to volume. This solution shows no more color than the straw yellow to greenish yellow color of the solution of the Standard solution similarly treated. Acceptance criteria: NMT 0.001% of N,N-dimethylaniline SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S Sample solution: 18 mg/mL (warm it, if necessary, to dissolve). Its specific rotation, determined photoelectrically at 325 nm, does not differ from that of the similarly prepared solution of USP Dextromethorphan Hydrobromide RS by more than 1.0%. PH 791: 5.26.5, in a solution (1 in 100) WATER DETERMINATION, Method I 921: 3.5%5.5% ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight containers. USP REFERENCE STANDARDS 11 USP Dextromethorphan Hydrobromide RS
Dextromethorphan Hydrobromide Oral Solution

(Comment on this Monograph)id=m24150=Dextromethorphan Hydrobromide Oral Solution=D-Monos.pdf) DEFINITION Dextromethorphan Hydrobromide Oral Solution contains NLT 90.0% and NMT 110.0% of the labeled amount of dextromethorphan hydrobromide (C18H25NO HBr H2O). IDENTIFICATION A. Transfer 50 mL of Oral Solution to a 250-mL separator, add 20 mL of water, 5 mL of 2.5 N sodium hydroxide, and 40 mL of solvent hexane, and shake thoroughly. Remove the solvent hexane layer, and filter through anhydrous sodium sulfate into a 150-mL beaker. Repeat the solvent hexane extraction, using two 40-mL portions and collecting the extracts in the beaker after filtering. Evaporate the combined extracts at 50 under nitrogen to dryness, and dissolve the residue in, and dilute with, 10 mL of chloroform: the solution is dextrorotatory (see Optical Rotation 781). Retain the chloroform solution for Identification test B. B. Evaporate the chloroform solution from Identification test A on a steam bath to dryness, dissolve the residue in 2 mL of 2 N sulfuric acid, and add 1 mL of a freshly prepared solution of mercuric nitrate (prepared by dissolving 700 mg of mercuric nitrate in 4 mL of water, adding 100 mg of sodium nitrate, mixing, and filtering): no red color is produced immediately, but after heating, a yellow to red color develops in about 15 min. ASSAY PROCEDURE Mobile phase: 0.007 M docusate sodium and 0.007 M ammonium nitrate in a mixture of acetonitrile and water (7:3). Adjust the solution with glacial acetic acid to a pH of 3.4. [NOTEDissolve the docusate sodium in the acetonitrile and water mixture before adding the ammonium nitrate.] Standard solution: 1 mg/mL of USP Dextromethorphan Hydrobromide RS in water. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, and dilute with Mobile phase to volume. Sample solution: Equivalent to 0.1 mg/mL of dextromethorphan hydrobromide from Oral Solution in water[NOTEUse a To Contain pipet.] Chromatographic system (See Chromatography 621, System Suitability.) Mode: LC Detector: UV 280 nm Column: 4.6-mm 25-cm; 5-m packing L1 Flow rate: 1 mL/min Injection size: 20 L System suitability Sample: Standard solution Suitability requirements Tailing factor: NMT 2.5 Relative standard deviation: NMT 2.0% Analysis Samples: Standard solution and Sample solution Calculate the percentage of C18H25NO HBr H2O in the volume of Oral Solution taken: Result = (rU/rS) (CS/CU) (Mr1/Mr2) 100 rU rS = peak response from the Sample solution = peak response from the Standard solution
USP 32
= concentration of USP Dextromethorphan Hydrobromide RS, on the anhydrous basis, in the Standard solution (mg/mL) = concentration of dextromethorphan CU hydrobromide in the Sample solution (mg/mL) = molecular weight of dextromethorphan Mr1 hydrobromide, 370.32 = molecular weight of anhydrous Mr2 dextromethorphan hydrobromide, 352.32 Acceptance criteria: 90.0%110.0% PERFORMANCE TESTS UNIFORMITY OF DOSAGE UNITS 905: Meets the requirements for Oral Solution packaged in Single-Unit Containers SPECIFIC TESTS DELIVERABLE VOLUME 698: Meets the requirements for Oral Solution packaged in Multiple-Unit Containers ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. USP REFERENCE STANDARDS 11 USP Dextromethorphan Hydrobromide RS CS
Official Monographs / Dextrose 75

ACIDITY Sample solution: Dissolve 5.0 g in 50 mL of carbon dioxidefree water. Add phenolphthalein TS. Analysis: Titrate with 0.020 N sodium hydroxide to the production of a distinct pink color. Acceptance criteria: NMT 0.30 mL is required for neutralization. WATER DETERMINATION, Method III 921: Dry it at 105 for 16 h: the hydrous form loses 7.5%9.5% of its weight, and the anhydrous form loses NMT 0.5% of its weight. OPTICAL ROTATION, Specific Rotation 781S: +52.6 to +53.2 Sample solution: 100 mg/mL in 0.012 N ammonium hydroxide DEXTRIN: Reflux 1 g of finely powdered Dextrose with 20 mL of alcohol: it dissolves completely. SOLUBLE STARCH, SULFITES: To a solution of 1 g in 10 mL of water, add 1 drop of iodine TS: the liquid is colored yellow. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. LABELING: Label it to indicate whether it is hydrous or anhydrous.
Dextrose
(Comment on this Monograph)id=m24180=Dextrose=DMonos.pdf)
Dextrose Injection
(Comment on this Monograph)id=m24230=Dextrose Injection=D-Monos.pdf) DEFINITION Dextrose Injection is a sterile solution of Dextrose in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of dextrose (C6H12O6 H2O). Dextrose Injection contains no antimicrobial agents. IDENTIFICATION Add a few drops of a solution (1 in 20) to 5 mL of hot alkaline cupric tartrate TS: a copious red precipitate of cuprous oxide is formed. ASSAY PROCEDURE Sample solution: Transfer a volume of Injection, containing 25 g of dextrose, to a 100-mL volumetric flask. Add 0.2 mL of 6 N ammonium hydroxide, and dilute with water to volume. Determine the angular rotation in a suitable polarimeter tube (see Optical Rotation 781). Analysis: Calculate the percentage, in g/100 mL, of C6H12O6 H2O in the portion of Injection taken: Result = A R (Mr1/Mr2) (100/M) A = 100 mm divided by the length of the polarimeter tube (mm) R = observed rotation () = molecular weight of dextrose monohydrate, Mr1 198.17 = molecular weight of anhydrous dextrose, 180.16 Mr2 = midpoint of the specific rotation range for M anhydrous dextrose, 52.9 Acceptance criteria: 95.0%105.0%
C6H12O6 H2O D-Glucose monohydrate [5996-10-1]; Anhydrous [50-99-7].
198.17 180.16
DEFINITION Dextrose is a sugar usually obtained by the hydrolysis of starch. It contains one molecule of water of hydration or is anhydrous. IDENTIFICATION Add a few drops of a solution (1 in 20) to 5 mL of hot alkaline cupric tartrate TS: a copious red precipitate of cuprous oxide is formed. IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% CHLORIDE AND SULFATE, Chloride 221: A 2.0-g portion shows no more chloride than corresponds to 0.50 mL of 0.020 N hydrochloric acid (0.018%). CHLORIDE AND SULFATE, Sulfate 221: A 2.0-g portion shows no more sulfate than corresponds to 0.50 mL of 0.020 N sulfuric acid (0.025%). ARSENIC, Method I 211: NMT 1 ppm HEAVY METALS 231: 4.0 g in 25 mL of water: NMT 5 ppm SPECIFIC TESTS COLOR OF SOLUTION: Dissolve 25 g in water to make 50.0 mL of solution: the solution has no more color than a solution prepared by mixing 1.0 mL of cobaltous chloride CS, 3.0 mL of ferric chloride CS, and 2.0 mL of cupric sulfate CS with water to make 10 mL, and diluting 3 mL of this solution with water to 50 mL. Make the comparison by viewing the solutions downward in matched color-comparison tubes against a white surface.
IMPURITIES Inorganic Impurities HEAVY METALS 231: Transfer a volume of Injection, equivalent to 4.0 g of dextrose, to a suitable vessel, and adjust the volume to 25 mL by evaporation or addition of water, as necessary: the limit is 0.0005C%, in which C is the labeled amount, in g, of C6H12O6 H2O/mL of Injection.
76
Dextrose / Official Monographs
USP 32
of 6 N ammonium hydroxide, and dilute with water to volume. Determine the angular rotation in a suitable polarimeter tube (see Optical Rotation 781). Analysis: Calculate the percentage, in g/100 mL, of C6H12O6 H2O in the portion of Injection taken: Result = A R (Mr1/Mr2) (100/M) = 100 mm divided by the length of the polarimeter tube (mm) R = observed rotation (degrees) Mr1 = molecular weight of dextrose monohydrate, 198.17 = molecular weight of anhydrous dextrose, 180.16 Mr2 = midpoint of the specific rotation range for M anhydrous dextrose, 52.9 SODIUM CHLORIDE Sample solution: Volume of Injection, equivalent to 90 mg of sodium chloride in 10 mL of glacial acetic acid and 75 mL of methanol. Add 3 drops of eosin Y TS. Analysis: Titrate, with shaking, with 0.1 N silver nitrate VS to a pink endpoint. Each mL of 0.1 N silver nitrate is equivalent to 5.844 mg of NaCl. Acceptance criteria: 95.0%105.0% IMPURITIES Organic Impurities PROCEDURE: LIMIT OF 5-HYDROXYMETHYLFURFURAL AND RELATED SUBSTANCES Sample solution: Volume of Injection, equivalent to 2.0 mg/mL of C6H12O6 H2O, in water Spectrometric conditions Cell: 1 cm Analytical wavelength: 284 nm Blank: Water Acceptance criteria: The absorbance is NMT 0.25. SPECIFIC TESTS PH 791: 3.26.5, determined on a portion diluted with water, if necessary, to a concentration of NMT 5% of dextrose BACTERIAL ENDOTOXINS TEST 85: It contains NMT 10.0 USP Endotoxin Units/g of dextrose. OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose glass or plastic containers. Glass containers are preferably of Type I or Type II glass. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, or where the label states that the Injection is not for direct injection but is to be diluted before use, the label alternatively may state the total osmolar concentration in mOsmol/mL. USP REFERENCE STANDARDS 11 USP Endotoxin RS A
Organic Impurities PROCEDURE: LIMIT OF 5-HYDROXYMETHYLFURFURAL AND RELATED SUBSTANCES Sample solution: Volume of Injection, equivalent to 4.0 mg/mL of C6H12O6 H2O, in water Spectrometric conditions Cell: 1 cm Analytical wavelength: 284 nm Blank: Water Acceptance criteria: The absorbance is NMT 0.25. SPECIFIC TESTS PH 791: 3.26.5, determined on a portion to which 0.30 mL of saturated potassium chloride solution has been added for each 100 mL and which previously has been diluted with water, if necessary, to a concentration of NMT 5% of dextrose PARTICULATE MATTER IN INJECTIONS 788: Meets the requirements BACTERIAL ENDOTOXINS TEST 85: It contains NMT 0.5 USP Endotoxin Unit/mL for Injections containing less than 5% of dextrose and NMT 10.0 USP Endotoxin Units/g for Injections containing between 5% and 70% of dextrose. [NOTEBefore analysis, dilute Injections containing more than 10% of dextrose to a concentration of 10% of dextrose.] OTHER REQUIREMENTS: It meets the requirements under Injections 1. ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in single-dose glass or plastic containers. Glass containers are preferably of Type I or Type II glass. LABELING: The label states the total osmolar concentration in mOsmol/L. Where the contents are less than 100 mL, or where the label states that the Injection is not for direct injection but is to be diluted before use, the label alternatively may state the total osmolar concentration in mOsmol/L. USP REFERENCE STANDARDS 11 USP Endotoxin RS
Dextrose and Sodium Chloride Injection

(Comment on this Monograph)id=m24300=Dextrose and Sodium Chloride Injection=D-Monos.pdf) DEFINITION Dextrose and Sodium Chloride Injection is a sterile solution of Dextrose and Sodium Chloride in Water for Injection. It contains NLT 95.0% and NMT 105.0% of the labeled amount of dextrose (C6H12O6 H2O) and of sodium chloride (NaCl). It contains no antimicrobial agents. IDENTIFICATION A. Add a few drops of a solution (1 in 20) to 5 mL of hot alkaline cupric tartrate TS: a copious red precipitate of cuprous oxide is formed. B. IDENTIFICATION TESTSGENERAL, Sodium 191 and Chloride 191: Meets the requirements ASSAY DEXTROSE Sample solution: Transfer a volume of Injection, containing 25 g of dextrose, to a 100-mL volumetric flask. Add 0.2 mL
USP 32
Official Monographs / Diatrizoate 77

Organic Impurities PROCEDURE: FREE AROMATIC AMINE Standard solution: Transfer 4 mL of water, 10 mL of 0.1 N sodium hydroxide, and 1.0 mL of Standard stock solution to a 50-mL volumetric flask. Sample solution: Transfer 1.0 g to a 50-mL volumetric flask, and add 5 mL of water and 10 mL of 0.1 N sodium hydroxide. Standard stock solution: Dissolve a suitable quantity of USP Diatrizoic Acid Related Compound A RS in 0.1 N sodium hydroxide. Use 0.2 mL of 0.1 N sodium hydroxide for each 5.0 mg of Standard, and dilute with water to obtain 500 g/mL. Blank: Transfer 5 mL of water and 10 mL of 0.1 N sodium hydroxide to a 50-mL volumetric flask. Analysis: Treat each flask as follows. Add 25 mL of dimethyl sulfoxide, insert the stopper, and mix by swirling gently. Chill in an ice bath in the dark for 5 min. [NOTEIn conducting the following steps, keep the flasks in the ice bath and in the dark as much as possible until all of the reagents have been added.] Slowly add 2 mL of hydrochloric acid and allow to stand for 5 min. Add 2 mL of sodium nitrite solution (1 in 50), and allow to stand for 5 min. Add 1 mL of sulfamic acid solution (2 in 25), shake, and allow to stand for 5 min. [CautionConsiderable pressure is produced.] Add 2 mL of a solution (1 in 1000) of N-(1-naphthyl)ethylenediamine dihydrochloride in dilute propylene glycol (7 in 10). Remove the flasks from the ice bath and from storage in the dark, and allow to stand in a water bath at 2225 for 10 min. Shake gently and occasionally during this period, releasing the pressure by loosening the stopper. Dilute with water to volume. Spectrometric conditions Mode: Spectrophotometry Analytical wavelength: 465 nm Cell: 1 cm Analysis: Within 5 min from the time of diluting the solutions in all three flasks to 50 mL, concomitantly determine the absorbances of the solutions. Acceptance criteria: The absorbance of the Sample solution is NMT that of the Standard solution (0.05%). SPECIFIC TESTS OPTICAL ROTATION, Specific Rotation 781S: 5.65 to 6.37 Sample solution: 100 mg/mL LOSS ON DRYING 731: Dry it at 105 for 4 h: it loses NMT 1.0% of its weight. IODINE AND IODIDE Sample solution: Transfer 2.0 g in a 50-mL centrifuge tube provided with a stopper, dilute with water to 24 mL, and shake to dissolve. Analysis: Add 5 mL of toluene and 5 mL of 2 N sulfuric acid, shake, and centrifuge: the toluene layer shows no red color. Add 1 mL of sodium nitrite solution (1 in 50), shake, and centrifuge: any red color in the toluene layer is not darker than that obtained when a mixture of 2.0 mL of potassium iodide solution (1 in 4000) and 22 mL of water is substituted for the solution under test. Acceptance criteria: NMT 0.02% of iodide ADDITIONAL REQUIREMENTS PACKAGING AND STORAGE: Preserve in well-closed containers. Store at 25, excursions permitted between 15 and 30. USP REFERENCE STANDARDS 11 USP Diatrizoic Acid RS USP Diatrizoic Acid Related Compound A RS
Diatrizoate Meglumine
(Comment on this Monograph)id=m24480=Diatrizoate Meglumine=D-Monos.pdf)
809.13 C11H9I3N2O4 C7H17NO5 Benzoic acid, 3,5-bis(acetylamino)-2,4,6-triiodo-, compd. with 1-deoxy-1-(methylamino)-D-glucitol (1:1); 1-Deoxy-1-(methylamino)-D-glucitol 3,5-diacetamido-2,4,6triiodobenzoate (salt) [131-49-7]. DEFINITION Diatrizoate Meglumine contains NLT 98.0% and NMT 102.0% of C11H9I3N2O4 C7H17NO5, calculated on the dried basis. IDENTIFICATION A. THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST 201 Solution A: Sodium hydroxide in methanol (0.8 in 1000) Standard solution: 1 mg/mL of USP Diatrizoic Acid RS in Solution A Sample solution: 1 mg/mL in Solution A Developing solvent system: Methanol, chloroform, and ammonium hydroxide (10:20:2) Analysis Samples: Standard solution and Sample solution Locate the spots using short-wavelength UV light. B. PROCEDURE Analysis: Heat 500 mg in a suitable crucible. Acceptance criteria: Violet vapors are evolved. ASSAY PROCEDURE Sample solution: Transfer 400 mg of Diatrizoate Meglumine to a glass-stoppered, 125-mL conical flask, add 30 mL of 1.25 N sodium hydroxide and 500 mg of powdered zinc, connect the flask to a reflux condenser, and reflux the mixture for 1 h. Cool the flask to room temperature, rinse the condenser with 20 mL of water, disconnect the flask from the condenser, and filter the mixture. Rinse the flask and the filter thoroughly, adding the rinsings to the filtrate. Add 5 mL of glacial acetic acid and 1 mL of tetrabromophenolphthalein ethyl ester TS. Analysis: Titrate with 0.05 N silver nitrate VS until the yellow precipitate just turns green. Each mL of 0.05 N silver nitrate is equivalent to 13.49 mg of C11H9I3N2O4 C7H17NO5. Acceptance criteria: 98.0%102.0% IMPURITIES Inorganic Impurities RESIDUE ON IGNITION 281: NMT 0.1% HEAVY METALS 231 Standard solution: To 2.0 mL of Standard Lead Solution (20 g of Pb) in a 50-mL color-comparison tube, add 5 mL of 1 N sodium hydroxide, and dilute with water to 40 mL. Sample solution: Dissolve 1.0 g of Diatrizoate Meglumine in 20 mL of water and 5 mL of 1 N sodium hydroxide, transfer the solution to a 50-mL color-comparison tube, and dilute with water to 40 mL. Analysis: To each of the tubes containing the Standard solution and the Sample solution add 10 mL of sodium sulfide TS, allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (0.002%).
78
Diatrizoate / Official Monographs
USP 32
Sample solution: In a 50-mL color-comparison tube, mix a volume of Injection, equivalent to 1.0 g of diatrizoate meglumine, with 5 mL of 1 N sodium hydroxide, and dilute with water to 40 mL. Analysis: To each of the tubes containing the Standard solution and the Sample solution add 10 mL of sodium sulfide TS, allow to stand for 5 min, and view downward over a white surface. Acceptance criteria: The color of the solution from the Sample solution is not darker than that of the solution from the Standard solution (0.002%). Organic Impurities PROCEDURE: FREE AROMATIC AMINE Sample solution: Transfer a volume of Injection equivalent to 1 g of diatrizoate meglumine, to a glass-stoppered, 50mL volumetric flask. Dilute with water to 5 mL, and add 10 mL of 0.1 N sodium hydroxide. Standard stock sol

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Official Monographs / Acarbose 1

(Comment on this Monograph)id=m115=Acarbose=AMonos.pdf)

Name Impurity Aa Impurity Bb Impurity Cc Impurity Dd Impurity Ee Impurity Ff Impurity Gg Impurity Hh

Acarbose / Official Monographs

Name Any individual unknown impurity

Official Monographs / Acebutolol 3

Acebutolol Hydrochloride Capsules

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Acepromazine Maleate Injection

Acepromazine / Official Monographs

Acepromazine Maleate Tablets

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C8H9NO2 Acetamide, N-(4-hydroxyphenyl)-; 4-Hydroxyacetanilide [103-90-2].

Acetaminophen / Official Monographs

Acetaminophen Oral Solution

Official Monographs / Acetaminophen 9

Acetaminophen for Effervescent Oral Solution

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Acetaminophen Oral Suspension

Official Monographs / Acetaminophen 11

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Acetaminophen Extended-Release Tablets

UNIFORMITY OF DOSAGE UNITS 905:

Meet the requirements

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Acetaminophen and Aspirin Tablets

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Acetaminophen, Aspirin, and Caffeine Tablets

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Official Monographs / Acetaminophen 23

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Official Monographs / Acetaminophen 25

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Official Monographs / Acetaminophen 27

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Official Monographs / Acetaminophen 29

Acetaminophen / Official Monographs

Acetaminophen, Chlorpheniramine Maleate, and Dextromethorphan Hydrobromide Tablets

Official Monographs / Acetaminophen 31

Acetaminophen and Codeine Phosphate Capsules

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Acetaminophen and Codeine Phosphate Oral Solution

Official Monographs / Acetaminophen 33

Acetaminophen and Codeine Phosphate Oral Suspension

Acetaminophen / Official Monographs

Acetaminophen and Codeine Phosphate Tablets

Official Monographs / Acetaminophen 35

Acetaminophen / Official Monographs

Official Monographs / Acetaminophen 37

Acetaminophen and Diphenhydramine Citrate Tablets

Acetaminophen / Official Monographs

Acetaminophen, Diphenhydramine Hydrochloride, and Pseudoephedrine Hydrochloride Tablets