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USP 36 Reagents / Reagents 1133
Reagents, Indicators and

Solutions
This section deals with the reagents and solutions re- Where a particular brand or source of a material or piece
quired in conducting the Pharmacopeial and the National of equipment, or the name and address of a manufacturer,
Formulary tests and assays. is mentioned, this identification is furnished solely for infor-
As is stated in the General Notices, listing of reagents, in- mational purposes as a matter of convenience, without im-
dicators, and solutions in the Pharmacopeia in no way im- plication of approval, endorsement, or certification.
plies that they have therapeutic utility; thus, any reference Atomic absorption and flame photometry require the use
to the USP in their labeling is to include the term reagent of a number of metal-ion standard solutions. While the indi-
or reagent grade. vidual monographs usually provide directions for prepara-
Reagents required in the tests and assays for the Pharma- tion of these solutions, use of commercially prepared stan-
copeial and National Formulary articles are listed in this sec- dardized solutions of the appropriate ions is permissible,
tion, generally with specifications appropriate to their in- provided that the analyst confirms the suitability of the solu-
tended uses. Exceptions to the latter include those reagents tions and has data to support their use.
for which corresponding specifications are presented in the Reagents are substances used either as such or as constit-
current edition of Reagent Chemicals, published by the uents of solutions.
American Chemical Society, and reagents for which specifi- Indicators are reagents used to determine the specified
cations could not be drafted in time for inclusion here. endpoint in a chemical reaction, to measure hydrogen-ion
Thus, where it is directed to Use ACS reagent grade, it is concentration (pH), or to indicate that a desired change in
intended that a grade meeting the corresponding specifica- pH has been effected. They are listed together with indica-
tions of the current edition of ACS Reagent Chemicals shall tor test papers.
be used. Where no such specifications exist, and where it is Buffer Solutions are referred to separately.
directed to Use a suitable grade, the intent is that a suita- Colorimetric Solutions, abbreviated CS, are solutions
ble reagent grade available commercially shall be used. Oc- used in the preparation of colorimetric standards for com-
casionally, additional test(s) augment the designation suita- parison purposes.
ble grade, as indicated in the text. Listed also are some, Test Solutions, abbreviated TS, are solutions of re-
but not all, reagents that are required only in determining agents in such solvents and of such definite concentrations
the quality of other reagents. For those reagents that are as to be suitable for the specified purposes.
not listed, satisfactory specifications are available in standard Volumetric Solutions, abbreviated VS and known also
reference works. as Standard Solutions, are solutions of reagents of known
In those instances in which a reagent required in a Phar- concentration intended primarily for use in quantitative de-
macopeial or National Formulary test or assay need not be terminations. Concentrations are usually expressed in terms
of analytical reagent quality, it suffices to refer to the mono- of normality.
graph for that article appearing in this Pharmacopeia or the WaterAs elsewhere in the Pharmacopeia, where
National Formulary or the current edition of the Food water, without qualification, is mentioned in the tests for
Chemicals Codex (FCC). In such cases it is to be understood reagents or in directions for preparing test solutions, etc.,
that the specifications are minimum requirements and that Purified Water (USP monograph) is always to be used. Car-
any substance meeting more rigid specifications for chemi- bon dioxide-free water is Purified Water that has been boiled
cal purity is suitable. vigorously for 5 minutes or more and allowed to cool while
Where the name of a reagent specified in a test or assay is protected from absorption of carbon dioxide from the at-
the same as the title of a USP or NF article, and it does not mosphere, or Purified Water that has a resistivity of not less
appear among the following Reagent Specifications, a sub- than 18 Mohm-cm. Deaerated water, for purposes other
stance meeting the requirements of the USP or NF mono- than dissolution and drug release testing, is Purified Water
graph is to be used (e.g., Benzocaine, USP; or Propylparaben, that has been treated to reduce the content of dissolved air
NF). However, reference is specifically made, under Reagent by suitable means, such as by boiling vigorously for 5 min-
Specifications, to a reagent bearing the name of a USP or NF utes and cooling or by the application of ultrasonic vibra-
article: (1) where there are requirements for a reagent in tion. Particle-free water is water that has been passed
addition to the USP or NF monograph requirements (e.g., through a 0.22-m filter.
Sodium Salicylate, USP; or Isopropyl Myristate, NF), (2) where Organic-free water is Purified Water that produces no sig-
a source other than the USP or NF monograph is specified nificantly interfering peaks when chromatographed as it is
(e.g., Lactose, ACS reagent; or Hydrochloric Acid, ACS rea- indicated in Identification, Control, and Quantification of Re-
gent), (3) where complete reagent specifications differ from sidual Solvents under Residual Solvents 467.
the USP or NF monograph standards (e.g., Calcium Lactate; Chromatographic Solvents and Carrier GasesThe
or Thymol), or (4) where a standard material is included chromatographic procedures set forth in the Pharmacopeia
among the reagent specifications (e.g., Calcium Carbonate, may require use of solvents and gases that have been espe-
primary standard; or Sodium Carbonate, primary standard). cially purified for such use. The purpose may be (a) to ex-
Reagents and solutions should be preserved in tight con- clude certain impurities that interfere with the proper con-
tainers made of resistant glass or other suitable material. duct of the test procedure, or (b) to extend the life of a
Directions for storage in light-resistant containers should be column by reducing the build-up of impurities on the col-
carefully observed. umn. Where solvents and gases are called for in chromato-
Stoppers and stopcocks brought into contact with sub- graphic procedures, it is the responsibility of the analyst to
stances capable of attacking or penetrating their surfaces ensure the suitability of the solvent or gas for the specific
may be given a protective coating of a thin film of a suita- use. Solvents and gases suitable for specific high-pressure or
ble lubricant unless specifically interdicted. other chromatographic uses are available as specialty prod-
ucts from various reagent supply houses, although there is
1134 Reagents / Reagents USP 36
no assurance that similar products from different suppliers uses of the solvents and gases and not for chromatographic
are of equivalent suitability in any given procedure. The rea- uses for which the especially purified specialty products may
gent specifications provided herein are for general analytical be required.
Reagents
For the purposes of the following specifications, these def-
Arsenic in Reagents
initions apply: A blank consists of the same quantities of the
same reagents treated in the same manner as the specimen
Select reagents for this test for a low arsenic content, so
under test. A control is a blank to which has been added the
that a blank test results in either no stain or one that is
limiting quantity of the substance being tested for, or is a
barely perceptible.
specified comparison solution prepared as directed in the
APPARATUSPrepare a generator by fitting a 1-hole rubber
particular test.
stopper into a wide-mouth bottle of about 60-mL capacity.
The values given in boldface type following chemical sym-
Through the perforation insert a vertical exit tube about
bols and formulas represent, respectively, atomic and molec-
12 cm in total length and 1 cm in diameter along the entire
ular weights of the substances concerned.
upper portion (for about 8 cm) and constricted at its lower
Color and turbidity comparisons are to be made in color-
extremity to a tube about 4 cm in length and about 5 mm
comparison tubes that are matched as closely as possible in
in diameter. The smaller portion of the tube should extend
internal diameter and in all other respects, as directed for
to just slightly below the stopper. Place washed sand or a
Visual Comparison under Spectrophotometry and Light-scatter-
pledget of purified cotton in the upper portion to about
ing 851. Such tubes frequently are called Nessler tubes.
3 cm from the top of the tube. Moisten the sand or cotton
In making visual comparisons of the densities of turbid
uniformly with lead acetate TS, and remove any excess or
fluids, compensate for differences in color, if necessary, by
adhering droplets of the latter from the walls of the tube.
viewing the turbidity through a column of water, the depth
Into the upper end of this tube fit a second glass tube
of which is determined by the volume specified in the indi-
12 cm in length, having an internal diameter of 2.5 to
vidual reagent specification. Place the water in color-com-
3 mm, by means of a rubber stopper. Just before running
parison tubes, and hold one of the tubes above the control
the test, place a strip of mercuric bromide test paper (see
tube and the other below the specimen tube.
under Indicator and Test Papers) in this tube, crimping the
Where an expression such as Retain the filtrate appears
upper end of the strip so that it will remain in position
it is to be understood, unless otherwise indicated, that the
about 2 cm above the rubber stopper. Clean and dry the
washings of the residue are not to be added to the filtrate
tube thoroughly each time it is used.
obtained. In the test heading, Calcium, magnesium, and R2O3
STANDARD ARSENIC SOLUTIONUse Standard Preparation pre-
precipitate, the expression R2O3 is intended to indicate the
pared as directed under Arsenic 211.
residue on ignition from compounds precipitated upon the
TEST PREPARATIONAdd 1 mL of sulfuric acid to 5 mL of a addition of ammonium hydroxide, such as Fe2O3 and Al2O3.
solution of the chemical substance (1 in 25), unless another
quantity is directed in the individual reagent specification.
Omit its addition entirely in the case of inorganic acids. Un-
GENERAL TESTS FOR REAGENTS
less especially directed otherwise, add 10 mL of sulfurous
acid. Evaporate the liquid in a small beaker, on a steam
The following general test methods are provided for the
bath, until it is free from sulfurous acid and has been re-
examination of reagents to determine their compliance with
duced to about 2 mL in volume. Dilute with water to 5 mL
the specifications of the individual reagents and are to be
to obtain the Test Preparation. Substances subjected to spe-
used unless it is otherwise directed in such specifications.
cial treatments specified in the individual reagent specifica-
tion may be used directly as the Test Preparation.
Boiling or Distilling Range for Reagents [NOTESolutions prepared by the dissolving of the chem-
ical substances in dilute acids are not considered to have
Use the following procedure for determining the boiling undergone special treatment.]
or distilling range of reagents, unless otherwise directed in
STANDARD STAINPlace in the generator bottle 5 mL of po-
the individual specifications:
tassium iodide TS, 2.0 mL of Standard Arsenic Solution, 5 mL
APPARATUSUse apparatus similar to that specified for Dis- of acid stannous chloride TS, and 28 mL of water. Add 1.5 g
tilling RangeMethod I 721, except that the distilling flask of granulated zinc (in No. 20 powder), and immediately in-
is to be of 250-mL capacity, to have a short neck, and to be sert the stopper containing the exit tube. Keep the genera-
connected to the condenser by means of a three-way con- tor bottle immersed in water at 25 during the period of the
necting tube fitted with standard-taper ground joints. test to moderate the reaction so that the stain will take the
form of a distinctive band to facilitate the comparison of
PROCEDUREPlace the distilling flask in an upright position
color intensity. When evolution of hydrogen has continued
in the perforation in the asbestos board, and connect it to
for 1 hour, remove the mercuric bromide test paper for
the condenser.
comparison. This stain represents 2 g of arsenic.
Measure 100 mL of the liquid to be tested in a graduated
cylinder, and transfer to the boiling flask together with PROCEDUREPipet into the generator bottle 5 mL of potas-
some device to prevent bumping. Use the cylinder as the sium iodide TS and 5 mL of the Test Preparation, and add
receiver for the distillate. Insert the thermometer, and heat 5 mL of acid stannous chloride TS. Set the apparatus aside
so as to distill at the rate of 3 mL to 5 mL per minute. Make at room temperature for a period of 10 minutes, then add
a preliminary trial, if necessary, to determine the adjustment 25 mL of water and 1.5 g of granulated zinc (in No.
for the proper rate of heating. Read the thermometer when 20 powder), and proceed as directed under Standard Stain.
about 20 drops have distilled and thereafter at volumes of Remove the mercuric bromide test paper, and compare the
distillate of 5, 10, 40, 50, 60, 90, and 95 mL. Continue the stain upon it with the Standard Stain: the stain produced by
distillation until the dry point is reached. the chemical tested does not exceed the standard stain in
The Boiling or Distilling Range is the interval between the length or in intensity of color, indicating not more than
temperatures when 1 mL and 95 mL, respectively, have 10 parts of arsenic per million parts of the substance being
distilled. tested. Since light, heat, and moisture cause the stain to
USP 36 Reagents / General Tests for Reagents 1135
fade rapidly, place the papers in clean, dry tubes, and make ity are available. The preferred type of flame photometer is
comparisons promptly. one that has a red-sensitive phototube, a multiplier photo-
tube, a monochromator, an adjustable slit-width control, a
INTERFERING CHEMICALSAntimony, if present in the sub-
selector switch, and a sensitivity control. Other types of
stance being tested, produces a gray stain. Sulfites, sulfides,
photometers may be used, provided the operator has
thiosulfates, and other compounds that liberate hydrogen
proved that the instrument will determine accurately the
sulfide or sulfur dioxide when treated with sulfuric acid must
amount of impurities permitted in the reagent to be tested.
be oxidized by means of nitric acid and then reduced by
The flame photometric procedures depend upon the use
means of sulfur dioxide as directed under Test Preparation
of semi-internal standards, and thus require both a Sample
before they are placed in the apparatus. Certain sulfur com-
Solution and a Control Solution. For the Sample Solution, a
pounds, as well as phosphine, give a bright yellow band on
specified weight of specimen is dissolved and diluted to a
the test paper. If sulfur compounds are present, the lead ace-
definite volume. For the Control Solution, the same amount
tate-moistened cotton or sand will darken. In that case, re-
of specimen is dissolved, the limiting amounts of the sus-
peat the operation as directed under Test Preparation upon a
pected impurities are added, and the solution is then di-
fresh portion of the solution being tested and use greater
luted to the same definite volume as the Sample Solution.
care in effecting the complete removal of the sulfurous acid.
The flame photometer is set as directed in the general pro-
In testing hypophosphites, observe special care to oxidize
cedures and then adjusted to give an emission reading as
completely the solution being tested as directed, otherwise
near 100% transmittance as is possible with the Control So-
the evolution of phosphine may result in a yellow stain
lution at the wavelength specified for the particular impurity
which may be confused with the orange-yellow color pro-
concerned. With the instrument settings left unchanged, the
duced by arsine. The stain produced by phosphine may be
emission from the Sample Solution is read at the same wave-
differentiated from that given by arsine by means of moist-
length and at a specified background wavelength. The back-
ening it with 6 N ammonium hydroxide. A stain caused by
ground reading is then used to correct the observed emis-
arsine becomes dark when so treated, but a stain produced
sion of the Sample Solution for the emission due to the
by phosphine does not materially change in color.
specimen and the solvent. The specimen being tested con-
tains less than the specified limit of impurity if the difference
Chloride in Reagents between the observed background and total emissions for
the Sample Solution is less than the difference between the
observed emissions for the Control Solution and the Sample
STANDARD CHLORIDE SOLUTIONDissolve 165.0 mg of dried
Solution at the wavelength designated for the particular
sodium chloride in water to make 1000.0 mL. This solution
impurity.
contains the equivalent of 0.10 mg of chlorine (Cl) in each
mL. CALCIUM IN REAGENTS
PROCEDURENeutralize, if alkaline, a solution of the quan- Standard Calcium SolutionDissolve 250 mg of calcium car-
tity of the reagent indicated in the test in 25 mL of water, bonate in a mixture of 20 mL of water and 5 mL of diluted
or a solution prepared as directed in the test, with nitric hydrochloric acid, and when solution is complete, dilute
acid, litmus paper being used as the indicator, and add with water to 1 L. This solution contains 0.10 mg of calcium
3 mL more of nitric acid. Filter the solution, if necessary, (Ca) per mL.
through a filter paper previously washed with water until
ProcedureUse the Sample Solution and the Control Solu-
the paper is free from chloride, and add 1 mL of silver ni-
tion prepared as directed in the individual test procedure.
trate TS. Mix, and allow to stand for 5 minutes protected
Set the slit-width control of a suitable flame photometer
from direct sunlight. Compare the turbidity, if any, with that
at 0.03 mm, and set the selector switch at 0.1. Adjust the
produced in a control made with the same quantities of the
instrument to give the maximum emission with the Control
same reagents as in the final test and a volume of Standard
Solution at the 422.7-nm calcium line, and record the trans-
Chloride Solution equivalent to the quantity of chloride (Cl)
mittance. Without changing any of the instrument settings,
permitted by the test. Adjust the two solutions with water
record the transmittance for the emission of the Sample So-
to the same volume before adding the silver nitrate TS, and
lution at 422.7 nm. Change the monochromator to the
compare the turbidities.
wavelength specified in the individual test procedure, and
In testing barium salts, neutralize, if alkaline, the solution
record the background transmittance for the background
containing the reagent, with nitric acid, and add only
emission of the Sample Solution: the difference between the
3 drops more of nitric acid. Conduct the remainder of the
transmittances for the Sample Solution at 422.7 nm and at
test as described previously.
the background wavelength is not greater than the differ-
In testing salts giving colored solutions, dissolve 2 g of the
ence between transmittances observed at 422.7 nm for the
reagent in 25 mL of water, and add 3 mL of nitric acid.
Sample Solution and the Control Solution.
Filter the solution, if necessary, through a filter paper previ-
POTASSIUM IN REAGENTS
ously washed with water, and divide the filtrate into two
Standard Potassium SolutionDissolve 191 mg of potassium
equal portions. Treat one portion with 1 mL of silver nitrate
chloride in a few mL of water, and dilute with water to 1 L.
TS, allow to stand for 10 minutes, and, if any turbidity is
Dilute a portion of this solution with water in the ratio of 1
produced, filter it through a washed filter paper until clear,
to 10 to obtain a concentration of 0.01 mg of potassium (K)
and use the filtrate as a blank. Treat the other portion with
per mL.
1 mL of silver nitrate TS, mix, and allow to stand for 5 min-
ProcedureUse the Sample Solution and the Control Solu- utes protected from direct sunlight. Compare the turbidity
tion prepared as directed in the individual test procedure. with that produced in the blank by the addition of a volume
[NOTEIn testing calcium salts, use an oxyhydrogen of Standard Chloride Solution equivalent to the quantity of
burner.] chloride (Cl) permitted in the test, both solutions being ad-
Set the slit-width control of a suitable flame photometer justed with water to the same volume.
equipped with a red-sensitive detector at 0.1 mm, unless
otherwise directed, and set the selector switch at 0.1. Adjust
Flame Photometry for Reagents the instrument to give the maximum emission with the Con-
trol Solution at the 766.5-nm potassium line, and record the
The use of flame photometric procedures to determine transmittance. Without changing any of the instrument set-
traces of calcium, potassium, sodium, and strontium is tings, record the transmittance for the emission of the Sam-
called for in some of the reagent specifications. The suitabil- ple Solution at 766.5 nm. Change the monochromator to
ity of such determinations depends upon the use of ade- 750 nm, and record the background transmittance for the
quate apparatus, and several instruments of suitable selectiv- background emission of the Sample Solution: the difference
1136 General Tests for Reagents / Reagents USP 36
between the transmittances for the Sample Solution at 766.5 tube add 10 mL of hydrogen sulfide TS, mix, and compare
nm and 750 nm is not greater than the difference between the colors by viewing through the color-comparison tube
transmittances observed at 766.5 nm for the Sample Solution downward against a white surface. The color in the test
and the Control Solution. specimen is not darker than that of the control.
If the solution of the reagent is prepared as in (b), use for
SODIUM IN REAGENTS
the control 10 mL of the solution, and add to it a volume of
Standard Sodium SolutionDissolve 254 mg of sodium chlo-
Standard Lead Solution equivalent to the amount of lead per-
ride in a few mL of water, and dilute with water to 1 L.
mitted in 2 g of the reagent. Dilute the remaining 30 mL of
solution (b) with water to 35 mL, and proceed as directed in
to 10 to obtain a concentration of 0.01 mg of sodium (Na)
the preceding paragraph, beginning with add diluted ace-
per mL.
tic acid, or ammonia TS, in the second sentence.
If the reagent to be tested for heavy metals is a salt of an
aliphatic organic acid, substitute 1 N hydrochloric acid for
the diluted acetic acid specified in the foregoing method.
Solution at the 589-nm sodium line, and record the trans- Insoluble Matter in Reagents
mittance. Without changing any of the instrument settings,
record the transmittance for the emission of the Sample Dissolve the quantity of reagent specified in the test in
Solutionat 589 nm. Change the monochromator to 580 nm, 100 mL of water, heat to boiling unless otherwise directed,
and record the background transmittance for the back- in a covered beaker, and warm on a steam bath for 1 hour.
ground emission of the Sample Solution: the difference be- Filter the hot solution through a tared sintered-glass crucible
tween the transmittances for the Sample Solution at 589 and of fine porosity. Wash the beaker and the filter thoroughly
580 nm is not greater than the difference between transmit- with hot water, dry at 105, cool in a desiccator, and
tances observed at 589 nm for the Sample Solution and the weigh.
Control Solution.
STRONTIUM IN REAGENTS
Loss on Drying for Reagents
Standard Strontium SolutionDissolve 242 mg of strontium
nitrate in a few mL of water, and dilute with water to 1 L.
Determine as directed under Loss on Drying 731.
to 10 to obtain a concentration of 0.01 mg of strontium (Sr)
per mL.
Nitrate in Reagents
STANDARD NITRATE SOLUTIONDissolve 163 mg of potassium
nitrate in water, add water to make 100 mL, and dilute
10 mL of this solution with water to 1 liter, to obtain a
solution containing the equivalent of 0.01 mg of NO3 per
Solution at the 460.7-nm strontium line, and record the
mL.
transmittance. Without changing any of the instrument set-
BRUCINE SULFATE SOLUTIONDissolve 600 mg of brucine sul-
tings, record the transmittance for the emission of the Sam-
fate in 600 mL of nitrate-free, dilute sulfuric acid (2 in 3)
ple Solution at 460.7 nm. Change the monochromator to
that previously has been cooled to room temperature, and
the wavelength specified in the individual test procedure,
dilute with the acid to 1 L. [NOTEPrepare the nitrate-free
and record the background transmittance for the back-
sulfuric acid by adding 4 parts of sulfuric acid to 1 part of
ground emission of the Sample Solution: the difference be-
water, heating the solution to dense fumes of sulfur trioxide,
tween the transmittances for the Sample Solution at 460.7
and cooling. Repeat the dilution and heating three or four
nm and at the background wavelength is not greater than
times.]
the difference between transmittances observed at 460.7
SAMPLE SOLUTIONTo the weight of sample specified in the
nm for the Sample Solution and the Control Solution.
individual reagent specification, dissolved in the designated
volume of water, add Brucine Sulfate Solution to make
50 mL. Heavy Metals in Reagents
CONTROL SOLUTIONTo a volume of Standard Nitrate Solu-
tion equivalent to the weight of nitrate (NO3) specified in STANDARD LEAD SOLUTIONUse Standard Lead Solution (see
the individual reagent specification, add the weight of sam- Heavy Metals 231). Each mL of this solution contains the
ple specified in the individual reagent specification and then equivalent of 0.01 mg of Pb.
add Brucine Sulfate Solution to make 50 mL.
PROCEDUREUnless otherwise directed, test for heavy met-
BLANK SOLUTIONUse 50 mL of Brucine Sulfate Solution. als as follows:
(a) If the heavy metals limit is 0.0005% (5 ppm), dis- PROCEDUREHeat the Sample Solution, Control Solution, and
solve 6.0 g of the specimen in water to make 42 mL. Blank Solution in a boiling water bath for 10 minutes, then
(b) If the heavy metals limit is 0.001% (10 ppm) or cool rapidly in an ice bath to room temperature. Adjust a
more, or in the event of limited solubility, use 4 g, and dis- suitable spectrophotometer to zero absorbance at 410 nm
solve in water to make 40 mL, warming, if necessary, to aid with the Blank Solution. Determine the absorbance of the
solution. Sample Solution, note the result, and adjust the instrument
For the control, transfer 7 mL of the solution from (a) to a to zero absorbance with the Sample Solution. Determine the
color-comparison tube, and add a volume of Standard Lead absorbance of the Control Solution: the absorbance reading
Solution equivalent to the amount of lead permitted in 4 g for the Sample Solution does not exceed that for the Control
of the reagent. Dilute with water to 35 mL, and add diluted Solution.
acetic acid, or ammonia TS, until the pH is about 3.5, deter-
mined potentiometrically, then dilute with water to 40 mL,
Nitrogen Compounds in Reagents and mix. Transfer the remaining 35 mL of the solution from
(a) to a color-comparison tube closely matching that used
for the control, and add diluted acetic acid, or ammonia TS, PROCEDUREUnless otherwise directed, test for nitrogen
until the pH is about 3.5, determined potentiometrically, compounds as follows: Dissolve the specified quantity of test
then dilute with water to 40 mL, and mix. Then to each specimen in 60 mL of ammonia-free water in a Kjeldahl flask
USP 36 Reagents / Reagent Specifications 1137
connected through a spray trap to a condenser, the end of PROCEDURE
which dips below the surface of 10 mL of 0.1 N hydrochlo-
Method INeutralize, if necessary, a solution of the quantity
ric acid. Add 10 mL of freshly boiled sodium hydroxide solu-
of the reagent or residue indicated in the test in 25 mL of
tion (1 in 10) and 500 mg of aluminum wire, in small
water, or a solution prepared as directed in the test, with
pieces, to the Kjeldahl flask, and allow to stand for 1 hour,
hydrochloric acid or with ammonia TS, litmus paper being
protected from loss of, and exposure to, ammonia. Distill
used as the indicator, and add 1 mL of 1 N hydrochloric
35 mL, and dilute the distillate with water to 50 mL. Add
acid. Filter the solution, if necessary, through a filter paper
2 mL of freshly boiled sodium hydroxide solution (1 in 10),
previously washed with water, and add 2 mL of barium
mix, add 2 mL of alkaline mercuric-potassium iodide TS, and
chloride TS. Mix, allow to stand for 10 minutes, and com-
again mix: the color produced is not darker than that of a
pare the turbidity, if any, with that produced in a control
control containing the amount of added N (as ammonium
containing the same quantities of the same reagents used in
chloride) specified in the individual test procedure.
the test and a quantity of Standard Sulfate Solution equiva-
lent to the quantity of sulfate (SO4) permitted in the test.
Adjust the two solutions with water to the same volume
Phosphate in Reagents
before adding the barium chloride TS.
Method IIHeat to boiling the solution, prepared as di-
STANDARD PHOSPHATE SOLUTIONDissolve 143.3 mg of dried
rected in the individual test procedure, or the filtrate desig-
monobasic potassium phosphate, KH2PO4, in water to make
nated in the procedure, and add 5 mL of barium chloride
1000.0 mL. This solution contains the equivalent of 0.10 mg
TS. Then digest the solution on a steam bath for 2 hours,
of phosphate (PO4) in each mL.
and allow to stand overnight. If any precipitate is formed,
PHOSPHATE REAGENT ADissolve 5 g of ammonium molyb-
filter the solution through paper, wash the residue with hot
date in 1 N sulfuric acid to make 100 mL.
water, and transfer the paper containing the residue to a
PHOSPHATE REAGENT BDissolve 200 mg of p-methylami- tared crucible. Char the paper, without burning, and ignite
nophenol sulfate in 100 mL of water, and add 20 g of so- the crucible and its contents to constant weight. Perform a
dium bisulfite. Store this reagent in well-filled, tightly stop- blank determination concurrently with the test specimen de-
pered bottles, and use within one month. termination, and subtract the weight of residue obtained
from that obtained in the test specimen determination to
PROCEDURE[NOTEThe tests with the specimen and the
obtain the weight of residue attributable to the sulfate con-
control are made preferably in matched color-comparison
tent of the specimen.
tubes.] Dissolve the quantity of the reagent specified in the
test, or the residue obtained after the prescribed treatment,
in 20 mL of water, by warming, if necessary, add 2.5 mL of
REAGENT SPECIFICATIONS
dilute sulfuric acid (1 in 7), and dilute with water to 25 mL.
(If preferable, the test specimen or the residue may be dis-
Absolute EtherSee Ethyl Ether, Anhydrous.
solved in 25 mL of approximately 0.5 N sulfuric acid.) Then
Absorbent CottonUse Purified Cotton (USP mono-
add 1 mL each of Phosphate Reagents A and B, mix, and
graph).
allow to stand at room temperature for 2 hours. Compare
Acetal, C6H14O2118.2Use a suitable grade.
any blue color produced with that produced in a control
Acetaldehyde (Ethanal; Acetic Aldehyde), CH3CHO
made with the same quantities of the same reagents as in
44.05 [75-07-0]Colorless liquid. Miscible with water
the test with the specimen, and a volume of Standard Phos-
and with alcohol. Use ACS reagent grade.
phate Solution equivalent to the quantity of phosphate (PO4)
Acetanilide (Phenylacetamide; Antifebrin), C8H9NO
designated in the reagent specifications.
135.16 [103-84-4]White, shiny crystals, usually in
scales, or a white, crystalline powder. Is stable in air. Freely
soluble in alcohol and in chloroform; soluble in boiling Residue on Ignition in Reagents
water, in ether, and in glycerin; slightly soluble in water.
Melting range 741: between 114 and 116. PROCEDUREUnless otherwise directed, determine the resi-
due on ignition as follows: Weigh accurately 1 to 2 g of the ReactionIts saturated solution is neutral to litmus.
substance to be tested in a suitable crucible that previously
Loss on drying 731Dry it over sulfuric acid for 2 hours:
has been ignited, cooled, and weighed. Ignite the sub-
it loses not more than 0.5% of its weight.
stance, gently and slowly at first and then at a more rapid
Residue on ignition (Reagent test): not more than
rate, until it is thoroughly charred, if organic in nature, or
0.05%.
until it is completely volatilized, if inorganic in nature. If the
Acetic Acid (6 N Acetic Acid)Use Acetic Acid (NF mono-
use of sulfuric acid is specified, cool the crucible, add the
graph) or prepare a suitable dilution of glacial acetic acid in
specified amount of acid, and ignite the crucible gently until
such a way as to obtain a final concentration of acetic acid
fumes no longer are evolved. Then ignite the crucible at
between 36.0% and 37.0%, by weight.
800 25, cool in a suitable desiccator, and weigh. If the
Acetic Acid, Diluted (1 N Acetic Acid)Dilute 60.0 mL of
use of sulfuric acid is not specified, the crucible need not be
glacial acetic acid with water to make 1000 mL.
cooled but can be ignited directly at 800 25 once the
Residue on evaporationEvaporate 50 mL on a steam
charring or volatilization is complete. Continue the ignition
bath, and dry the residue at 105 for 2 hours: the residue
until constant weight is attained, unless otherwise specified.
weighs not more than 1 mg (0.002%).
Conduct the ignition in a well-ventilated hood, but pro-
tected from air currents, and at as low a temperature as is Chloride (Reagent test)Five mL shows not more than
possible to effect the complete combustion of the carbon. A 0.01 mg of Cl (2 ppm).
muffle furnace may be used, if desired, and its use is recom-
Sulfate (Reagent test, Method I )Ten mL shows not
mended for the final ignition at 800 25.
more than 0.5 mg of SO4 (50 ppm).
Heavy metals (Reagent test)Evaporate 20 mL on a
steam bath to dryness. Add to the residue 2 mL of the acid,
Sulfate in Reagents
dilute with water to 25 mL, and add 10 mL of hydrogen
sulfide TS: any brown color produced is not darker than that
STANDARD SULFATE SOLUTIONDissolve 181.4 mg of potas-
sium sulfate (dried at 105 for 2 hours) in water to make
1000 mL. This solution contains the equivalent of 0.10 mg
of sulfate (SO4) per mL.
1138 Reagent Specifications / Reagents USP 36
of a control containing 0.04 mg of added Pb and 2 mL of Melting range 741When previously dried at 110 in a
the diluted acetic acid (2 ppm). capillary tube for 1 hour, it melts between 149 and 152.
Acetic Acid, Glacial, CH3COOH60.05 [64-19-7]
ReactionA solution (1 in 10) is neutral to litmus.
Use ACS reagent grade.
Residue on ignition (Reagent test): negligible, from
Acetic Anhydride (Acetic Oxide; Acetyl Oxide),
200 mg.
(CH3CO)2O102.09 [108-24-7]Use ACS reagent
Solubility in alcoholA solution of 500 mg in 5 mL of al-
grade.
cohol is complete and colorless.
Acetone (Propanone; Dimethylformaldehyde),
Percent of acetyl (CH3CO)Weigh accurately about CH3COCH358.08 [67-64-1]Use ACS reagent grade.
400 mg, previously dried at 105 for 3 hours, and dissolve in [NOTEFor UV spectrophotometric determinations, use
15 mL of water in a glass-stoppered conical flask. Add ACS reagent grade Acetone Suitable for Use in UV
40.0 mL of 0.1 N sodium hydroxide VS, and heat on a Spectrophotometry.]
steam bath for 30 minutes. Insert the stopper, allow to cool, Acetone, Anhydrous, CH3COCH358.08Use ACS rea-
add phenolphthalein TS, and titrate the excess alkali with gent grade Acetone.
0.1 N sulfuric acid VS. Determine the exact normality of the Acetone, NeutralizedTo a suitable quantity of acetone
0.1 N sodium hydroxide by titrating 40.0 mL after it has add 2 or 3 drops of phenophthalein TS and a sufficient
been treated in the same manner as in the test. Each mL of amount of 0.02 or 0.01 N sodium hydroxide to produce a
0.1 N sodium hydroxide is equivalent to 4.305 mg of faint pink color. Prepare neutralized acetone just prior to
CH3CO. Between 23.2% and 24.2% is found. use.
Acetonitrile (Methyl Cyanide; Cyanomethane), CH3CN Percent of chlorine (Cl)Weigh accurately about 400 mg,
41.05 [75-05-8]Use ACS reagent grade. previously dried at 105 for 3 hours, and dissolve in 50 mL
Acetonitrile, SpectrophotometricUse ACS reagent of water in a glass-stoppered, 125-mL flask. Add with agita-
grade, which meets also the requirements of the following tion 30.0 mL of 0.1 N silver nitrate VS, then add 5 mL of
test. nitric acid and 5 mL of nitrobenzene, shake, add 2 mL of
ferric ammonium sulfate TS, and titrate the excess silver ni- Spectral purityMeasure in a 1-cm cell between 250 nm
trate with 0.1 N ammonium thiocyanate VS: each mL of 0.1 and 280 nm, with a suitable spectrophotometer, against air
N silver nitrate is equivalent to 3.545 mg of Cl. Between as the blank: its absorbance is not more than 0.01.
19.3% and 19.8% of Cl is found. Acetophenone (Phenylethanone; Phenyl Methyl Ketone),
3-Acetylthio-2-methylpropanoic Acid, C6H10O3S CH3COC6H5120.15 [98-86-2]Liquid. Slightly soluble
162.21Use a suitable grade. in water, freely soluble in alcohol and in ether.
[NOTEA suitable grade is available as -(Acetylmer-
Melting range 741: between 19 and 20.
capto)isobutyric Acid, catalog number 39059, from Senn
Refractive index 831: about 1.534 at 20.
Chemicals AG www.sennchem.com.]
Specific gravity 841: about 1.03.
N-Acetyl-L-tyrosine Ethyl Ester, C13H17NO4251.28De-
p-Acetotoluidide, C9H11NO149.19 [103-89-9]
termine the suitability of the material as directed in the As-
White to off-white powder.
say under Chymotrypsin (USP monograph).
AssayInject an appropriate volume into a gas chromat- Acrylic Acid (2-Propenoic Acid; Vinylformic Acid),
ograph (see Chromatography 621) equipped with a flame- C3H4O272.06 [79-10-7]Colorless liquid. Miscible with
ionization detector, helium being used as the carrier gas. water, with alcohol, and with ether.
The following conditions have been found suitable: a 0.25-
AssayInject an appropriate specimen into a gas chro-
mm 30-m capillary column coated with a 1-m layer of
matograph (see Chromatography 621), equipped with a
phase G2; the injection port temperature is maintained at
flame-ionization detector, helium being used as the carrier
230; the detector temperature is maintained at 300; and
gas. The following conditions have been found suitable: a
the column temperature is maintained at 130 and pro-
0.25-mm 30-m capillary column coated with a 1-m layer
grammed to rise 10 per minute to 280. The area of the
of phase G2; the injection port temperature is maintained at
C9H11NO peak is not less than 98.5% of the total peak area.
Melting range 741: between 145 and 151. the column temperature is maintained at 50 and pro-
Acetylacetone (2,4-Pentanedione; Diacetylmethane), grammed to rise 10 per minute to 200. The area of the
C5H8O2100.12 [123-54-6]Clear, colorless to slightly C3H4O2 peak is not less than 99% of the total peak area.
yellow, flammable liquid. Soluble in water; miscible with al-
Refractive index 831: between 1.419 and 1.423 at
cohol, with chloroform, with acetone, with ether, and with
20.
glacial acetic acid.
Activated AluminaSee Alumina, Activated.
AssayNot less than 98% of C5H8O2, a suitable gas chro- Activated CharcoalSee Charcoal, Activated.
matograph equipped with a flame-ionization detector being Activated Magnesium SilicateSee Magnesium Silicate,
used and helium being used as the carrier gas. The follow- Activated.
ing conditions have been found suitable: a 3-mm 1.83-m Adamantane, C10H16136.23 [281-23-2]
stainless steel column containing 10% phase G43 on sup-
port S1A; the injection port and detector temperatures are
Adenine Sulfate, (C5H5N5)2 H2SO4 2H2O404.36
maintained at 250 and 310, respectively; the column tem-
White crystals or crystalline powder. Melts, after drying at
perature is programmed to rise 8 per minute, from 50 to
110, at about 200 with some decomposition. One g dis-
220.
solves in about 160 mL of water; less soluble in alcohol. Sol-
Refractive index 831: between 1.4505 and 1.4525, at uble in solutions of sodium hydroxide. It is not precipitated
20. from solution by iodine TS or mercuric-potassium iodide TS,
Acetyl Chloride, CH3COCl78.50 [75-36-5]Clear, but a precipitate is produced with trinitrophenol TS.
colorless liquid. Is decomposed by water and by alcohol.
Miscible with benzene and with chloroform. Use ACS rea-
100 mg.
gent grade.
WaterDry it at 105 to constant weight: it loses not
more than 10.0% of its weight.
Acetylcholine Chloride (Trimethylethanaminium Chloride;
Adipic Acid (Hexanedioic Acid; 1,4-Butanedicarboxylic Acid),
Acecoline), [CH3COOCH2CH2N(CH3)3]Cl181.66
C6H10O4146.14 [124-04-9]Colorless to white, crystal-
[60-31-1]White, crystalline powder. Very deliquescent;
line powder. Slightly soluble in water and in cyclohexane;
very soluble in water; freely soluble in alcohol.
soluble in alcohol, in methanol, and in acetone; practically
insoluble in benzene and in petroleum benzin.
AssayWeigh accurately about 0.3 g, and dissolve in hol, gasoline, isopropanol, terpineol, benzene, castor oil, ac-
50 mL of alcohol. Add 25 mL of water, mix, and titrate with etone, nicotine, aniline dyes, ether, cadmium iodide, pyri-
0.5 N sodium hydroxide VS to a pH of 9.5. Perform a blank dine bases, sulfuric acid, kerosene, and diethyl phthalate.
determination, and make any necessary correction. Each mL Use a suitable grade.
of 0.5 N sodium hydroxide is equivalent to 36.54 mg of Alcohol, DilutedUse Diluted Alcohol (NF monograph).
C6H10O4. Not less than 98% is found. Alcohol, IsobutylSee Isobutyl Alcohol.
Alcohol, IsopropylSee Isopropyl Alcohol.
Melting range 741: between 151 and 155, but the
Alcohol, MethylSee Methanol.
range between beginning and end of melting does not ex-
Alcohol, NeutralizedTo a suitable quantity of alcohol
ceed 2.
add 2 or 3 drops of phenolphthalein TS and just sufficient
AgarUse Agar (NF monograph). When used for bacteri-
0.02 N or 0.1 N sodium hydroxide to produce a faint pink
ological purposes, it is to be dried to a water content of not
color. Prepare neutralized alcohol just prior to use.
more than 20%.
Alcohol, n-PropylSee n-Propyl Alcohol.
Agarose [9012-36-6]Polysaccharide consisting of 1,3-
Alcohol, Secondary ButylSee Butyl Alcohol, Secondary.
linked -D-galactopyranose and 1,4-linked 3,6-anhydro--L-
Alcohol, Tertiary ButylSee Butyl Alcohol, Tertiary.
galactopyranose. Use a suitable grade.
Aldehyde DehydrogenaseA white powder. One mg
AirHelium Certified StandardA mixture of 1.0% air in
contains not less than 2 enzyme activity units.
industrial grade helium. It is available from most suppliers of
specialty gases. AssayTransfer about 20 mg, accurately weighed, to a
Albumin Bovine Serum [9048-46-8]Almost colorless to 200-mL volumetric flask, dissolve in 1 mL of water, dilute
faintly yellow powder. Not less than 95% pure. Solubility, with an ice-cold solution of bovine serum albumin (1 in
40 mg in 1 mL of water. Molecular weight is approximately 100) to volume, and mix. Use this solution as the Assay
66,000. Use a suitable grade. Store between 2 and 8. preparation. Dissolve 3.3 g of potassium pyrophosphate,
Alcohol, Ethanol, Ethyl Alcohol, C2H5OH46.07 15 mg of dithiothreitol, and 40 mg of edetate disodium in
[64-17-5]Use a suitable grade with a content of NLT 70 mL of water, adjust with citric acid monohydrate solution
92.3% and NMT 93.8%, by weight, corresponding to NLT (2.1 in 10) to a pH of 9.0 0.1, dilute with water to
94.9% and NMT 96% by volume, at 15.56. 100 mL, and mix to obtain a pH 9.0 buffer. Dissolve an ac-
Alcohol, 70 Percent, 80 Percent, and 90 PercentPre- curately weighed quantity of -nicotinamide adenine
pare by mixing alcohol and water in the proportions given, dinucleotide (-NAD) in water to obtain a -NAD solution
the measurements being made at 25. having a known concentration of about 20 mg per mL. Pi-
pet 0.1 mL of the Assay preparation into a 1-cm spectropho-
tometric cell. Pipet 0.1 mL of water into a second 1-cm
Relative Proportions
spectrophotometric cell to provide the reagent blank. Add
Volume in
2.5 mL of pH 9.0 buffer, 0.2 mL of -NAD solution, and
mL of
0.1 mL of pyrazole solution (0.68 in 100) to each cell, and
Percent by Alcohol,
mix. Stopper the cells, and allow to stand for 2 minutes at
Volume of Specific 94.9% v/v,
25 1. Add 0.01 mL of acetaldehyde solution (0.3 in 100)
C2H5OH at Gravity Alcohol, Water, Required
to each cell, and mix. Stopper the cells, and determine the
15.56 at 25 mL mL for 100 mL
absorbance of the solution obtained from the Assay prepara-
70 0.884 38.6 15 73.7
tion at a wavelength of 340 nm, using the solution obtained
80 0.857 45.5 9.5 84.3
from the reagent blank as the reference. Calculate the
90 0.827 51 3 94.8 change, A, in absorbance per minute for the solution ob-
tained from the Assay preparation, starting at the point
The proportions of alcohol and water taken to prepare
when the absorbance and time relationship becomes linear.
these or any other percentage (v/v) solutions may be deter-
One enzyme activity unit is defined as the amount of en-
mined as follows. Calculate the amount, in mL, of water to
zyme that oxidizes 1 mol of acetaldehyde per minute when
be mixed with 100 mL of alcohol taken by the formula:
the test is conducted under the conditions described herein.
Calculate the enzyme activity units in each mg of aldehyde
[94.9(d/c) 0.8096]100
dehydrogenase taken by the formula:
in which 94.9 is the percentage (v/v) of C2H5OH in alcohol,
[(2.91)(200)/(6.3)(0.1)(1000)](A/W)
0.8096 is the specific gravity of 94.9% alcohol, d is the
specific gravity, obtained from the Alcoholometric Table (see
in which A is as defined above and W is the weight, in g,
Reference Tables), of the solution containing c% (v/v) of
of aldehyde dehydrogenase taken.
C2H5OH, and 100 is the volume, in mL, of alcohol taken.
Alkaline Phosphatase EnzymeSee Phosphatase Enzyme,
Alcohol, Absolute, C2H5OH46.07Use ACS reagent
Alkaline.
grade Ethyl Alcohol, Absolute.
AlkylphenoxypolyethoxyethanolA nonionic surfactant.
Alcohol, Aldehyde-freeDissolve 2.5 g of lead acetate in
Use a suitable grade.
5 mL of water, add the solution to 1000 mL of alcohol con-
[NOTEA suitable grade is available commercially as Tri-
tained in a glass-stoppered bottle, and mix. Dissolve 5 g of
ton X-100 from Sigma-Aldrich, www.sigma-aldrich.com.]
potassium hydroxide in 25 mL of warm alcohol, cool the
Alpha-Chymotrypsin25 kDa [9004-07-3]Use a suit-
solution, and add it slowly, without stirring, to the alcohol
able salt-free grade for protein sequencing.
solution of lead acetate. After 1 hour shake the mixture vig-
[NOTEA suitable grade is available as catalog number
orously, allow it to stand overnight, decant the clear liquid,
4423 from www.sigma-aldrich.com.]
and recover the alcohol by distillation.
Alpha-Cyclodextrin Hydrate (Alpha-Schardinger Dextrin;
Alcohol, AmylSee Amyl Alcohol.
Cyclohexaamylose), C36H60O30 xH2O [51211-51-9]Use a
Alcohol, Dehydrated (Absolute Alcohol), C2H5OH
suitable grade with a content of NLT 98%.
46.07Use ACS reagent grade Ethyl Alcohol, Absolute.
Alcohol, Dehydrated IsopropylSee Isopropyl Alcohol,
22729 from www.acros.com.]
Dehydrated.
Alpha-(2-(methylamino)ethyl)benzyl alcoholUse a
Alcohol, DenaturatedIt is ethyl alcohol to which has
suitable grade.
been added some substance or substances which, while al-
AlphanaphtholSee 1-Naphthol.
lowing the use of the alcohol in most applications, renders
Alprenolol Hydrochloride, C15H23NO2 HCl285.8
it entirely unfit for consumption as a beverage. The most
[13707-88-5]Use a suitable grade.
common denaturants used, either alone or in combination,
Alum (Ammonium Alum, Aluminum Ammonium Sulfate),
are the following: methanol, camphor, aldehol, amyl alco-
AlNH4(SO4)2 12H2O453.33 [7784-26-1]Large, color-
less crystals or crystalline fragments or a white powder. Sol- Nitrogen content (Reagent test)Determine by the
uble in 7 parts of water and in about 0.5 part of boiling Kjeldahl method, using a test specimen previously dried at
water; insoluble in alcohol. Use ACS reagent grade. 105 for 2 hours: between 18.4% and 18.8% of N is found,
Ammonium AlumSee Alum. corresponding to not less than 98.5% of C2H5NO2.
AluminaSee Aluminum Oxide, Acid-washed.
Insoluble matter (Reagent test): not more than 1 mg,
Alumina, Activated (Aluminum Oxide), [1344-28-1]Use
from 10 g (0.01%).
a suitable grade.
Residue on ignition (Reagent test): not more than 0.05%.
Alumina, Anhydrous (Aluminum Oxide; Alumina specially
Chloride (Reagent test)One g shows not more than
prepared for use in chromatographic analysis) [1344-28-1]A
0.1 mg of Cl (0.01%).
white or practically white powder, 80- to 200-mesh. It does
Sulfate (Reagent test, Method I)Two g shows not more not soften, swell, or decompose in water. It is not acid-
than 0.1 mg of SO4 (0.005%). washed. Store it in well-closed containers.
Aluminon (Aurin Tricarboxylic Acid, [tri]Ammonium Salt), Heavy metals (Reagent test): 0.001%, 5 mL of 1 N hydro-
C22H23N3O9473.43 [569-58-4]Yellowish-brown, glassy chloric acid VS being used to acidify the solution of the test
powder. Freely soluble in water. Use ACS reagent grade. specimen.
Aluminum, AlAt. Wt. 26.98154 [7429-90-5]Use
Iron 241One g, dissolved in 47 mL of water contain-
ACS reagent grade, which also meets the requirements of
ing 3 mL of hydrochloric acid, shows not more than
the following test.
0.01 mg of Fe (0.001%).
ArsenicPlace 750 mg in a generator bottle (see Arsenic 4-Aminoantipyrine, C11H13N3O203.24 [83-07-8]
in Reagents under General Tests for Reagents), omitting the Light yellow, crystalline powder. A 500-mg portion dissolves
pledget of cotton. Add 10 mL of water and 10 mL of so- completely in 30 mL of water and yields a clear solution.
dium hydroxide solution (3 in 10), and allow the reaction to
proceed for 30 minutes: not more than a barely perceptible
p-Aminobenzoic AcidSee Para-aminobenzoic Acid.
stain is produced on the mercuric bromide test paper.
2-Aminobenzonitrile (Anthranilonitrile), C7H6N2118.14
Aluminum Oxide, Acid-Washed (Alumina specially pre-
[1885-29-6]Use 2-Aminobenzonitrile 98%.
pared for use in chromatographic analysis) [1344-28-1]
White or practically white powder or fine granules. Very hy-
4-Amino-6-chloro-1,3-benzenedisulfonamide,
groscopic. Store in tight containers.
C6H8ClN3O4S2285.73 [121-30-2]White powder. Insol-
pH of SlurryThe pH of a well-mixed slurry of 5 g in
uble in water and in chloroform; soluble in ammonia TS.
150 mL of ammonia-free and carbon dioxide-free water, af-
Residue on ignition (Reagent test): not more than 2 mg
ter 10 minutes standing, is between 3.5 and 4.5.
from 2 g (0.1%).
Loss on ignitionWeigh accurately about 1 g, and ignite,
AbsorbanceA 1 in 200,000 solution in methanol exhib-
preferably in a muffle furnace at 800 to 825, to constant
its absorbance maxima at about 223 nm, 265 nm, and 312
weight: it loses not more than 5.0% of its weight.
nm. Its absorptivity (see Spectrophotometry and Light-Scatter-
SilicaFuse 500 mg with 10 g of potassium bisulfate for
ing 851) at 265 nm is about 64.0.
1 hour in a platinum crucible, cool, and dissolve in hot
4-Amino-2-chlorobenzoic Acid, C6H3Cl(NH2)(COOH)
water: not more than a small amount of insoluble matter
171.58 [2457-76-3]White crystals or white, crystalline
remains.
powder.
Suitability for chromatographic adsorptionDissolve 50 mg
of o-nitroaniline in benzene to make 50.0 mL. Dilute 10 mL
2-Amino-5-chlorobenzophenone, C13H10ClNO231.68
of the resulting solution with benzene to 100.0 mL, and mix
[719-59-5]Use USP 2-Amino-5-chlorobenzophenone RS.
(Solution A).
7-Aminodesacetoxycephalosporanic Acid, C8H10N2O3S
Weigh quickly about 2 (0.005) g of specimen in a glass-
214.2Light yellow powder.
stoppered weighing bottle, and rapidly transfer it to a dry,
Ordinary impurities 466
glass-stoppered test tube. Add 20.0 mL of Solution A, insert
Test solution: 1 N ammonium hydroxide.
the stopper, shake vigorously for 3 minutes, and allow to
settle. Standard solution: 1 N ammonium hydroxide.
Pipet 10 mL of the clear supernatant into a 100-mL volu-
Eluant: 0.5 N sodium chloride.
metric flask, dilute with benzene to volume, and mix (Solu-
Visualization: 1.
tion B).
2-Aminoethyl DiphenylborinateSee Diphenylborinic
Determine the absorbances of Solutions A and B at 395
Acid, Ethanolamine Ester.
nm, with a suitable spectrophotometer, using benzene as
1-(2-Aminoethyl)piperazine, C6H15N3129.20
the blank. Calculate the quantity, in mg, adsorbed per g of
[140-31-8]Viscous, colorless liquid.
test specimen by the formula:
AssayInject an appropriate specimen into a suitable gas
chromatograph (see Chromatography 621) equipped with
[2(1 AB/AA)]/W
a flame-ionization detector, helium being used as the carrier
in which AA and AB are the absorbances of Solutions A and B,
0.25-mm 30-m capillary column coated with G2. The in-
respectively; and W is the weight, in g, of the aluminum
jection port temperature is maintained at 280; the column
oxide. Not less than 0.3 mg of o-nitroaniline is adsorbed for
temperature is maintained at 180 and programmed to rise
each g of the aluminum oxide.
10 per minute to 280 and held there for 10 minutes. The
Aluminum Potassium Sulfate, AlK(SO4)2 12H2O
detector temperature is maintained at 300. The area of the
474.39 [10042-67-1]Use ACS reagent grade.
main peak is not less than 97% of the total peak area.
Amaranth, C20H11N2Na3O10S3604.48 [915-67-3]A
deep brown or dark reddish-brown fine powder. Use a suita- Refractive index 831: between 1.4978 and 1.5010 at
ble grade. 20.
Aminoacetic Acid (Glycine), NH2CH2COOH75.07
[56-40-6]White, crystalline powder. Very soluble in water;
slightly soluble in alcohol.
Aminoguanidine Bicarbonate (Aminoguanidine Hydrogen Sparingly soluble in cold water; freely soluble in hot water,
Carbonate), CH6N4 H2CO3136.11 [2582-30-1]White in alcohol, and in ether.
powder.
AssayDissolve about 1.5 g, accurately weighed, in
AssayDissolve about 34 mg, accurately weighed, in about 400 mL of water in a 500-mL volumetric flask, dilute
50 mL of glacial acetic acid. Titrate with 0.1 N perchloric with water to volume, and mix. Transfer 25.0 mL of this
acid VS, determining the endpoint potentiometrically. Per- solution to an iodine flask, add 50.0 mL of 0.1 N bromine
form a blank determination, and make any necessary correc- VS, dilute with 50 mL of water, add 5 mL of hydrochloric
tions. Each mL of 0.1 N perchloric acid is equivalent to acid, and immediately insert the stopper in the flask. Shake
13.61 mg of CH6N4 H2CO3. Not less than 98.5% is found. for 1 minute, allow to stand for 2 minutes, and add 5 mL of
potassium iodide TS through the slightly loosened stopper.
Melting point 741: about 170, with decomposition.
Shake thoroughly, allow to stand for 5 minutes, remove the
2-Aminoheptane (2-Heptylamine; 1-Methylhexylamine),
stopper, and rinse it and the neck of the flask with 20 mL of
C7H17N115.22 [123-82-0]Use a suitable grade with a
water, adding the rinsing to the flask. Titrate the liberated
content of not less than 99%.
iodine with 0.1 N sodium thiosulfate VS, adding 3 mL of
N-Aminohexamethyleneimine (N-Aminohomopiperidine,
starch TS as the endpoint is approached. From the volume
1-Aminohomopiperidine), C6H14N2114.19 [5906-35-4]
of 0.1 N sodium thiosulfate used, calculate the volume, in
Colorless liquid.
mL, of 0.1 N bromine consumed by the test specimen. Each
mL of 0.1 N bromine is equivalent to 1.819 mg of C6H7NO:
not less than 99.5% is found.
0.25-mm 30-m capillary column coated with G2. The in- Loss on drying 731Dry it over calcium chloride for
jection port temperature is maintained at 180; the column 4 hours: the loss in weight is negligible.
Residue on ignition (Reagent test): negligible, from 2 g.
10 per minute to 230 and then maintained at 230 for
p-Aminophenol (p-Hydroxyaniline), C6H7NO109.13
5 minutes. The detector temperature is maintained at 300.
[123-30-8]Fine, yellowish, crystalline powder. Slightly sol-
The area of the main peak is not less than 95% of the total
uble in water and in alcohol. Use a suitable grade with a
peak area.
Refractive index 831: between 1.4840 and 1.4860 at 3-Amino-1-propanol, H2N(CH2)3OH75.11
20. [156-87-6]Liquid.
4-Amino-3-hydroxy-1-naphthalenesulfonic Acid,
Boiling range (Reagent test): between 184 and 188.
C10H9NO4S239.25 [116-63-2]Light purple powder.
Refractive index 831: between 1.461 and 1.463 at 20.
3-Aminopropionic Acid (-Alanine), NH2CH2CH2COOH
8-Amino-6-methoxyquinoline (6-Methoxy-8-aminoquin-
89.09 [107-95-9]Use a suitable grade.
oline), C10H10N2O174.2 [90-52-8]Use a suitable grade
3-Aminosalicylic Acid, C7H7NO3153.14 [570-23-0]
with a content of not less than 98.0%.
Tan-grey powder. Use a suitable grade with a content of not
[NOTEA suitable grade is available from www.3bmedical-
less than 97%.
systems.com, catalog number 3B3-002598.]
Ammonia Detector TubeA fuse-sealed glass tube so
1,2,4-Aminonaphtholsulfonic Acid, C10H9NO4S
designed that gas may be passed through it and containing
239.25White to slightly brownish pink powder. Sparingly
suitable absorbing filters and support media for the indica-
soluble in water.
tor bromophenol blue.
SensitivenessDissolve 100 mg in 50 mL of freshly pre-
Measuring range: 5 to 70 ppm.
pared sodium bisulfite solution (1 in 5), warming if neces-
[NOTEAvailable from Draeger Safety, Inc.,
sary to effect solution, and filter. Add 1 mL of the filtrate to
www.draeger.com, or from Gastec Corp., www.gastec.co.jp,
a solution prepared by adding 2 mL of dilute sulfuric acid (1
distributed in the USA by www.nextteq.com.]
in 6) and 1 mL of Phosphate Reagent A (see Reagent test) to
Ammonia Solution, DilutedUse Ammonia TS.
20 mL of a 1 in 100 dilution of Standard Phosphate Solution
Ammonia Water, 25 Percent [1336-21-6]Use a suitable
(see Reagent test): a distinct blue color develops within
grade.
5 minutes.
Ammonia Water, Stronger (Ammonium Hydroxide)
Solubility in sodium carbonate solutionDissolve 100 mg
[1336-21-6]Use ACS reagent grade Ammonium Hydrox-
in 3 mL of sodium carbonate TS, and add 17 mL of water:
ide.
not more than a trace remains undissolved.
Ammonium Acetate, NH4C2H3O277.08 [631-61-8]
Residue on ignition (Reagent test)To 1 g add 0.5 mL of Use ACS reagent grade.
sulfuric acid, and ignite at 800 25 to constant weight: the Ammonium Bicarbonate (Ammonium Hydrogen
residue weighs not more than 5 mg (0.5%). Carbonate), NH4HCO379.06 [1066-33-7]Use a suitable
grade with a content of NLT 99.0%.
Sulfate (Reagent test, Method I)Heat 500 mg with a
Ammonium Bisulfate (Ammonium Hydrogen Sulfate),
mixture of 25 mL of water and 2 drops of hydrochloric acid
NH4HSO4115.11 [7803-63-6]White crystals. Freely
on a steam bath for 10 minutes. Cool, dilute with water to
soluble in water; practically insoluble in alcohol, in acetone,
200 mL, and filter: 20 mL of the filtrate shows not more
and in pyridine.
than 0.25 mg of SO4 (0.5%).
2-Aminophenol (o-Aminophenol; 2-Hydroxyaniline), AssayDissolve about 300 mg, accurately weighed, in
C6H7NO109.13 [95-55-6]Off-white powder. Use a 50 mL of a mixture of water and alcohol (25:25). Titrate
suitable grade with a content of not less than 99%. with 0.1 N sodium hydroxide VS, determining the endpoint
m-Aminophenol (3-Amino-1-Hydroxybenzene), C6H7NO potentiometrically. Perform a blank determination and make
109.13 [591-27-5]Cream-colored to pale yellow flakes. any necessary correction. Each mL of 0.1 N sodium hydrox-
ide is equivalent to 11.51 mg of NH4HSO4. Not less than a stream of carbon dioxide through the solution to remove
98% is found. any excess sulfur dioxide, then cool, and titrate with 0.1 N
Ammonium Bromide, NH4Br97.94 [12124-97-9] potassium permanganate VS. Each mL of 0.1 N potassium
Use ACS reagent grade. permanganate consumed is equivalent to 11.7 mg of
Ammonium Carbonate (Hartshorn Salt) [506-87-6]Use NH4VO3. Not less than 98.0% is found.
ACS reagent grade.
Solubility in ammonium hydroxideDissolve 1 g in a mix-
Ammonium Chloride (Salmiac), NH4Cl53.49
ture of 3 mL of ammonium hydroxide and 50 mL of warm
[12125-02-9]Use ACS reagent grade.
water: the solution is clear and colorless.
Ammonium Citrate, Dibasic (Citric Acid Diammonium
CarbonateTo 500 mg add 1 mL of water and 2 mL of
Salt), (NH4)2HC6H5O7226.18 [3012-65-5]Use ACS re-
diluted hydrochloric acid: no effervescence is produced.
agent grade.
ChlorideDissolve 250 mg in 40 mL of hot water, add
Ammonium Dihydrogen PhosphateSee Ammonium
2 mL of nitric acid, and allow to stand for 1 hour. Filter, and
Phosphate, Monobasic.
to the filtrate add 0.5 mL of silver nitrate TS: any turbidity
Ammonium Fluoride, NH4F37.04 [12125-01-8]
produced does not exceed that of a blank containing
0.5 mg of added Cl (0.2%).
Ammonium Formate (Formic Acid Ammonium Salt),
SulfateDissolve 500 mg in 50 mL of hot water, and add CH5NO263.06 [540-69-2]Use a suitable grade.
2 mL of diluted hydrochloric acid and 1.5 g of hydroxyla- Ammonium Hydroxide (Ammonium Aqueous),
mine hydrochloride. Heat at 60 for 3 minutes, filter, cool, [1336-21-6]Use ACS reagent grade.
and add to the filtrate 2 mL of barium chloride TS: no tur- Ammonium Hydroxide, 6 NPrepare by diluting 400 mL
bidity or precipitate is produced within 30 minutes. of Ammonia Water, Stronger (see Reagents section) with
Amyl Acetate (Isoamyl Acetate), CH3CO2C5H11130.18 water to make 1000 mL.
[2308-18-1]Clear, colorless liquid. Slightly soluble in Ammonium Molybdate, (NH4)6Mo7O24 4H2O1235.86
water. Miscible with alcohol, with amyl alcohol, with ben- [12054-85-2]Use ACS reagent grade.
zene, and with ether. Ammonium Nitrate, NH4NO380.04 [6484-52-2]
Use ACS reagent grade. Specific gravity 841: about 0.87.
Ammonium Oxalate, (NH4)2C2O4 H2O142.11
Boiling range (Reagent test, Method I): not less than
90%, between 137 and 142.
Ammonium Persulfate (Ammonium Peroxydisulfate),
Solubility in diluted alcoholA 1.0-mL portion dissolves in
(NH4)2S2O8228.20 [7727-54-0]Use ACS reagent
20 mL of diluted alcohol to form a clear solution.
grade Ammonium Peroxydisulfate.
AcidityAdd 5.0 mL to 40 mL of neutralized alcohol, and,
Ammonium Phosphate, Dibasic (Diammonium Hydrogen
if the pink color is discharged, titrate with 0.10 N sodium
Phosphate), (NH4)2HPO4132.06 [7783-28-0]Use ACS
hydroxide: not more than 0.20 mL is required to restore the
reagent grade.
pink color (about 0.02% as CH3COOH).
Ammonium Phosphate, Monobasic (Ammonium
Dihydrogen Phosphate), NH4H2PO4115.03 WaterA 5-mL portion gives a clear solution with 5 mL
[7722-76-1]Use ACS reagent grade. of carbon disulfide.
Ammonium Pyrrolidinedithiocarbamate (1-pyrrolidine- Amyl Alcohol (Isoamyl Alcohol), C5H11OH88.15
carbodithioic acid, ammonium salt), C5H12N2S2164.29 [598-75-4]Use ACS reagent grade Isopentyl Alcohol.
[5108-96-3]Use a suitable grade. tert-Amyl Alcohol, C5H12O88.15 [75-85-4]Clear,
Ammonium Reineckate (Reinecke Salt), NH4 colorless, flammable, volatile liquid.
[Cr(NH3)2(SCN)4] H2O354.44 [13573-16-5]Dark red
crystals or red, crystalline powder. Moderately soluble in
Boiling range (Reagent test): not less than 95%, be-
cold water; more soluble in hot water. Gradually decom-
tween 100 and 103.
poses in solution.
Residue on evaporationEvaporate 50 mL (40 g) on a
SensitivenessDissolve 50 mg in 10 mL of water. Add
steam bath, and dry at 105 for 1 hour: the residue weighs
0.2 mL of the solution to 1 mL of a solution of 10 mg of
not more than 1.6 mg (0.004%).
choline chloride in 20 mL of water, and shake gently: a dis-
Acids and estersDilute 20 mL with 20 mL of alcohol,
tinct precipitate forms within 5 to 10 seconds.
add 5.0 mL of 0.1 N sodium hydroxide VS, and reflux gen-
Ammonium Sulfamate, NH4OSO2NH2114.13
tly for 10 minutes. Cool, add 2 drops of phenolphthalein TS,
and titrate the excess sodium hydroxide with 0.1 N hydro-
Ammonium Sulfate, (NH4)2SO4132.14
chloric acid VS: not more than 0.75 mL of the 0.10 N so-
dium hydroxide is consumed, correction being made for the
Ammonium Thiocyanate (Ammonium Rhodanide),
amount consumed in a blank (0.06% as amyl acetate).
NH4SCN76.12 [1762-95-4]Use ACS reagent grade.
AldehydesShake 5 mL with 5 mL of potassium hydrox- Ammonium Vanadate (Ammonium Metavanadate),
ide solution (30 in 100) in a glass-stoppered cylinder for NH4VO3116.98 [7803-55-6]White, crystalline pow-
5 minutes, and allow to separate: no color develops in either der. Slightly soluble in cold water; soluble in hot water and
layer. in dilute ammonia TS.
-AmylaseUse a suitable grade. It can be from vegetal
AssayWeigh accurately about 500 mg, transfer to a
or animal or microbiological origin.
suitable container, add 30 mL of water and 2 mL of dilute
(E)-Anethole (1-Methoxy-4-(1-propenyl)benzene),
sulfuric acid (1 in 4), swirl to dissolve, and pass sulfur diox-
C10H12O148.20 [4180-23-8]Use a suitable grade of
ide gas through the solution until reduction is complete and
transisomer.
the solution is bright blue in color. Boil gently while passing
Anhydrous AluminaSee Alumina, Anhydrous. and convert it completely to the chloride form by stirring
Anhydrous Barium ChlorideSee Barium Chloride, Anhy- with 150 mL of hydrochloric acid (5 in 100) for not less
drous. than 30 minutes. Decant the acid, and wash the resin in the
Anhydrous Calcium ChlorideSee Calcium Chloride, An- same manner with distilled water until the wash water is
hydrous. neutral to litmus.
Anhydrous Cupric SulfateSee Cupric Sulfate, Anhydrous. Transfer 5 to 7 mL of the regenerated resin to a glass
Anhydrous Dibasic Sodium PhosphateSee Sodium filtering crucible, and remove only the excess surface water
Phosphate, Dibasic, Anhydrous. by very careful suction filtration. Transfer the conditioned,
Anhydrous Magnesium PerchlorateSee Magnesium dried resin to a tared weighing bottle, and weigh. Dry in a
Perchlorate, Anhydrous. vacuum oven at 100 to 105 and at a pressure of 50 mm
Anhydrous Magnesium SulfateSee Magnesium Sulfate, of mercury for 16 hours. Transfer from the vacuum oven to
Anhydrous. a desiccator, and cool to room temperature. Reweigh. The
Anhydrous MethanolSee Methanol, Anhydrous. loss in weight is between 50% and 65%.
Anhydrous Potassium CarbonateSee Potassium Car-
Total new volume capacityTransfer 2.5 to 3 mL of the
bonate, Anhydrous.
conditioned, undried (See Moisture content, above) resin to
Anhydrous Sodium AcetateSee Sodium Acetate, Anhy-
a 5-mL graduated cylinder, and fill it with water. Remove
drous.
any air bubbles from the resin bed with a stainless steel
Anhydrous Sodium CarbonateSee Sodium Carbonate,
wire, and settle the resin to its minimum volume by tapping
Anhydrous.
the graduated cylinder. Record the volume of the resin.
Anhydrous Sodium SulfateSee Sodium Sulfate, Anhy-
Transfer the resin with 100 mL of water to a 250-mL flask.
drous.
Add 2 mL of sulfuric acid, heat to 70 to 80, and hold at
Anhydrous Sodium SulfiteSee Sodium Sulfite, Anhy-
that temperature for 5 minutes with occasional stirring (do
drous.
not boil). Cool to room temperature, and add 2.5 mL of
Aniline, C6H5NH293.13 [62-53-3]Use ACS reagent
nitric acid (1 in 2), 2 mL of ferric ammonium sulfate TS, and
grade.
0.20 mL of 0.1 N ammonium thiocyanate. Titrate with 0.1
Aniline Blue (Certified Biological Aniline Blue)
N silver nitrate VS until the solution turns colorless, and add
[8004-91-9]A water-soluble dye consisting of a mixture of
a measured excess (1 to 5 mL). Heat to boiling to coagulate
the tri-sulfonates of triphenylpararosaniline and of
the silver chloride precipitate. Cool to room temperature,
diphenylrosaniline.
add 10 mL of nitrobenzene, shake vigorously, and titrate the
Aniline Sulfate, C12H14N2 H2SO4284.33
excess silver nitrate with 0.1 N ammonium thiocyanate VS.
Anion-Exchange Resin, Chloromethylated Polystyrene- (net mL AgNO3 N)/(mL of resin) = mEq/mL
DivinylbenzeneStrongly basic, cross-linked resin contain-
ing quaternary ammonium groups. It consists of small, The total exchange capacity of the regenerated, wet resin is
moist, yellow beads having a characteristic amine odor. It is more than 1.0 mEq per mL.
available in the chloride form which can be converted to
Wet screen analysisThe purpose of this test is to identify
the hydroxide form by regeneration with sodium hydroxide
properly the mesh size of the resin. To obtain an accurate
solution (1 in 4). For satisfactory regeneration a contact
screen analysis requires special apparatus and technique.
time of about 25 minutes is required, after which it must be
Add 150 mL of resin to 200 mL of distilled water in an
washed with water until neutral. Suitable for use in column
appropriate bottle, and allow it to stand at least 4 hours to
chromatography.
completely swell the resin.
[NOTEA suitable resin is Amberlite IRA-400, available
Transfer by means of a graduated cylinder 100 mL of set-
from Sigma-Aldrich, www.sigma-aldrich.com.]
tled and completely swollen resin to the top screen of a
Anion-Exchange Resin, Strong, Lightly Cross-Linked, in
series (20-, 50-, 100-mesh) of 20.3-cm brass screens. Thor-
the Chloride FormUse a suitable grade.
oughly wash the resin on each screen with a stream of dis-
[NOTEA suitable resin is AG 1-X4, catalog number
tilled water until the resin is completely classified, collecting
140-1331, produced by BioRad Laboratories, www.bio-rad.
the wash water in a suitable container. Wash the beads re-
com.]
maining on the respective screens back into the 100-mL cyl-
Anion-Exchange Resin, Styrene-Divinylbenzene
inder, and record the volume of settled resin on each
Strongly basic, cross-linked resin containing quaternary am-
screen: not less than 80% of the resin is between 50- and
monium groups and about 8% of divinylbenzene. It is avail-
100-mesh.
able in the chloride form in the 50- to 100-, 100- to 200-,
p-Anisaldehyde (4-Methoxybenzaldehyde), C8H8O2
and 200- to 400-mesh sizes. It can be converted to the
136.15 [123-11-5]Clear, colorless liquid.
hydroxide form by regeneration with a sodium hydroxide
Boiling temperature: 248.
solution (5 in 100). Insoluble in water, in methanol, and in
Density: between 1.119 and 1.123.
acetonitrile. Suitable for use in column chromatography.
[NOTEA suitable resin is Dowex 1X8, produced by Dow Refractive index 831: between 1.5725 and 1.5730 at
Chemical Co. (www.dow.com) and available through 20.
Sigma-Aldrich (www.sigma-aldrich.com).] p-Anisidine, C7H9NO123.06 [104-94-9]Brown
Anion-Exchange Resin, 50- to 100-Mesh, Styrene- crystals. Use a suitable grade.
DivinylbenzeneStrongly basic, cross-linked resin contain- Anisole, CH3OC6H5108.14 [100-66-3]Colorless liq-
ing quaternary ammonium groups and about 4% of uid.
divinylbenzene. It consists of tan-colored beads that may be
AssayInject an appropriate specimen (about 0.5 L)
relatively free flowing. It is available in the chloride form
into a suitable gas chromatograph (see Chromatography
which can be converted to the hydroxide form by regenera-
621) equipped with a flame-ionization detector, nitrogen
tion with a sodium hydroxide solution (5 in 100). For satis-
being used as the carrier gas. The following conditions have
factory regeneration a contact time of at least 30 minutes is
been found suitable: a 30-m capillary column is coated with
required after which it must be washed free of excess alkali.
phase G3; the injection port and detector temperatures are
Insoluble in water, in methanol, and in acetonitrile. Suitable
maintained at 140 and 300, respectively; the column tem-
for use in column chromatography.
perature is maintained at 70 and programmed to rise 10
[NOTEA suitable resin is Dowex 1X4, available from
per minute to 170. The area of the anisole peak is not less
Sigma-Aldrich, www.sigma-aldrich.com.]
than 99% of the total peak area.
Moisture content of fully regenerated and expanded resin
Refractive index 831: 1.5160 at 20.
Transfer 10 to 12 mL of the resin (as received) to a flask,
Anthracene, C14H10178.23 [120-12-7]White to off- pears the endpoint has been reached. Each mL of 0.1 N
white crystals or platelets. Darkens in sunlight. Insoluble in sodium thiosulfate is equivalent to 14.95 mg of SbCl5: not
water; sparingly soluble in alcohol, in benzene, and in chlo- less than 99.0% of SbCl5 is found.
roform.
Sulfate (Reagent test, Method II)Dissolve 4.3 mL (10 g)
Melting range 741: between 215 and 218. in the minimum volume of hydrochloric acid, dilute with
Anthrone, C14H10O194.23 [90-44-8]Use ACS rea- water to 150 mL, neutralize with ammonium hydroxide, and
gent grade. filter. To the filtrate add 2 mL of hydrochloric acid: the solu-
Anti-D ReagentThe reagent can be monoclonal (low tion, 10 mL of barium chloride TS being used, yields not
protein) or polyclonal (high-protein) and must be obtained more than 1.3 mg of residue, correction being made for a
from manufacturers or suppliers licensed by the Center for complete blank test (0.005%).
Biologics Evaluation and Research, Food and Drug Adminis-
ArsenicAdd 10 mL of a recently prepared solution of
tration. The use of reagents from an unlicensed manufac-
20 g of stannous chloride in 30 mL of hydrochloric acid to
turer or supplier may invalidate the results. Note that this
100 mg of specimen dissolved in 5 mL of hydrochloric acid.
reagent is different from Anti-D (Rho) Reagent. Consult man-
Mix, transfer to a color-comparison tube, and allow to stand
ufacturers package insert to ensure that the reagent is suita-
for 30 minutes. Any color in the solution of the specimen
ble for the Weak Anti-D test and does not contain other
should not be darker than that in a control containing
antibodies that will react when antihuman immunoglobulin
0.02 mg of arsenic (As), which has been treated in the same
is added.
manner as the test specimen, when viewed downward over
[NOTEThere are many manufacturers and suppliers of
a white surface (0.02% of As).
these reagents that are licensed by the Center for Biologics
Substances not precipitated by hydrogen sulfide (as SO4)
Evaluation and Research, Food and Drug Administration.
Dissolve 0.90 mL (2 g) in 5 mL of hydrochloric acid, and di-
Some examples of licensed manufacturers or suppliers are
lute with 95 mL of water. Precipitate the antimony com-
the following: Gamma Biologics, Houston, TX; and Ortho
pletely with hydrogen sulfide, allow the precipitate to settle,
Diagnostics, Raritan, NJ.]
and filter, being careful not to transfer much of the precipi-
Anti-D (Rho) ReagentThe reagent can be monoclonal
tate to the filter paper. (Retain the precipitate.) To 50 mL of
or polyclonal and must be obtained from manufacturers or
the filtrate, add 0.5 mL of sulfuric acid, evaporate in a tared
suppliers licensed by the Center for Biologics Evaluation and
porcelain crucible to dryness, and ignite at 800 25 for
Research, Food and Drug Administration for use in
15 minutes. (Retain the residue.) The weight of the ignited
microplate tests. The use of reagents from an unlicensed
residue should not be more than 0.0010 g greater than the
manufacturer or supplier may invalidate the results. Note
weight obtained in a complete blank test (0.10%).
that this reagent is different from Anti-D Reagent. Consult
Iron 241To the residue from the test for Substances
manufacturers package insert to ensure that it is Anti-D
not precipitated by hydrogen sulfide add 2 mL of hydrochloric
(Rho) Reagent and not Anti-D Reagent.
acid and 5 drops of nitric acid, and evaporate on a steam
bath to dryness. Take up the residue in 2 mL of hydrochloric
acid, and dilute with water to 47 mL: the solution shows
not more than 0.01 mg of Fe (0.001%).
Other heavy metals (as Pb)Dissolve the precipitate on the following: Gamma Biologics, Houston, TX; and Ortho
the filter paper from the test for Substances not precipitated Diagnostics, Raritan, NJ.]
by hydrogen sulfide, with 75 mL of a solution containing 6 g Antifoam ReagentA 10% siliconeglycol emulsion,
of sodium sulfide and 4 g of sodium hydroxide dissolved in white in appearance. Nonionic but miscible with cool water.
and diluted with water to 100 mL. Collect the filtrate in the [NOTEA suitable grade is available as Antifoam
original flask containing the remainder of the sulfide precipi- Reagent, catalog number 2210, from Dow Corning Corpo-
tate. Warm the solution to dissolve the soluble sulfides, and ration, www.dowcorning.com.]
allow the insoluble sulfides to settle. Filter, wash thoroughly Antihuman Globulin ReagentThe reagent can be poly-
with hydrogen sulfide TS, and dissolve any precipitate re- specific or anti-immunoglobulin (Anti-IgG) and must be ob-
maining on the filter paper with 10 mL of hot diluted hy- tained from manufacturers or suppliers licensed by the
drochloric acid. Dilute the filtrate with water to 50 mL. Neu- Center for Biologics Evaluation and Research, Food and
tralize a 25-mL portion of this solution with 1 N sodium Drug Administration. The use of reagents from an unli-
hydroxide, and add 1 mL of 1 N acetic acid and 10 mL of censed manufacturer or supplier may invalidate the results.
hydrogen sulfide TS. Any brown color should not exceed [NOTEThere are many manufacturers and suppliers of
that produced by 0.05 mg of lead ion in an equal volume of these reagents that are licensed by the Center for Biologics
solution containing 1 mL of 1 N acetic acid and 10 mL of Evaluation and Research, Food and Drug Administration.
hydrogen sulfide TS (0.005%). Some examples of licensed manufacturers or suppliers are
Antimony Trichloride (Antimonous Chloride), SbCl3 the following: Gamma Biologics, Houston, TX; and Ortho
228.12 [10025-91-9]Use ACS reagent grade. Diagnostics, Raritan, NJ.]
Antithrombin IIIAntithrombin III human (heparin Antimony Pentachloride, SbCl5299.02
cofactor, factor IIa inhibitor, and factor Xa inhibitor) is a ser- [7647-18-9]Clear, reddish-yellow, oily, hygroscopic, caus-
ine protease inhibitor. It is a glycoprotein having a molecu- tic liquid. Fumes in moist air and solidifies by absorption of
lar weight of 58,000 Da. one molecule of water. Is decomposed by water; soluble in
One Antithrombin III Unit is the amount found in 1 mL of dilute hydrochloric acid and in chloroform. Boils at about
normal human plasma. The potency of antithrombin III is 92 at a pressure of 30 mm of mercury and has a specific
not less than 4.0 Antithrombin III Units per mg of protein gravity of about 2.34 at 25.
when tested in the presence of heparin. It exhibits 90% [CAUTIONAntimony pentachloride causes severe burns, and
homogeneity when tested by SDS-PAGE. the vapor is hazardous.]
Antithrombin III for test or assay purposes contains no
Assay (SbCl5)Accurately weigh a glass-stoppered,
detectable heparin. Test as follows. To a solution containing
125-mL flask, quickly introduce about 0.3 mL of the test
1 Antithrombin III Unit per mL, add 1 L of toluidine blue
specimen, and reweigh. Dissolve with 20 mL of diluted hy-
solution. In the presence of heparin, the color changes from
drochloric acid (1 in 5), and add 10 mL of potassium iodide
blue to purple.
solution (1 in 10) and 1 mL of carbon disulfide. Titrate the
Aprobarbital, C10H14N2O3210.23 [77-02-1]Fine,
liberated iodine with 0.1 N sodium thiosulfate VS. The
white crystalline powder. Slightly soluble in cold water; solu-
brown color will gradually disappear from the solution, and
ble in alcohol, in chloroform, and in ether.
the last traces of free iodine will be collected in the carbon
disulfide, giving a pink color. When this pink color disap-
AssayDissolve about 200 mg, previously dried at 105 Barium Chloride DihydrateUse Barium Chloride.
for 2 hours and accurately weighed, in 20 mL of dimethyl- Barium Hydroxide, Ba(OH)2 8H2O315.46
formamide in a 100-mL conical flask. Add 4 drops of thymol [12230-71-6]Use ACS reagent grade.
blue solution (1 in 200 in methanol), and titrate with 0.1 N Barium Nitrate, Ba(NO3)2261.34 [10022-31-8]Use
lithium methoxide VS using a 10-mL buret, a magnetic stir- ACS reagent grade.
rer, and a cover for the flask to protect against atmospheric Basic FuchsinSee Fuchsin, Basic.
carbon dioxide. Perform a blank determination, and make Beclomethasone C22H29ClO5408.92 [4419-39-0]
any necessary correction. Each mL of 0.1 N lithium methox- Use a suitable grade with a content of not less than 99%.
ide is equivalent to 21.02 mg of C10H14N2O3. Between Beef ExtractA concentrate from beef broth obtained by
98.5% and 101.0% of C10H14N2O3 is found. extraction from fresh, sound, lean beef by means of cooking
with water and evaporating the broth at a low temperature,
usually in vacuum, until a thick, pasty residue results. Yel-
Arsenazo III Acid, C22H18As2N4O14S2776.38
lowish brown to dark brown, slightly acid, pasty mass hav-
[1668-00-4]Brown powder. Stable in air. Store at room
ing an agreeable meat-like odor. Store it in tight, light-resis-
temperature in a dry area.
tant containers.
Melting temperature 741: greater than 320.
For the following tests, prepare a test solution by dissolv-
Arsenic Trioxide, As2O3197.84 [1327-53-3]Use
ing 25 g in water to make 250 mL of a practically clear and
ACS reagent grade.
practically sediment-free solution.
[NOTEArsenic Trioxide of a quality suitable as a primary
Assay for nitrogen content of alcohol-soluble substances
standard is available from the National Institute of Standards
Place a portion of the alcohol filtrate and washings remain-
and Technology, Office of Standard Reference Materials,
ing from the test for Alcohol-insoluble substances, corre-
www.nist.gov, as standard sample No. 83.]
sponding to 1 g of the alcohol-soluble solids, in a 500-mL
L-Asparagine (L-2-Aminosuccinamic Acid),
Kjeldahl flask. Add about 10 g of powdered potassium sul-
COOHCH(NH2)CH2CONH2 H2O150.13 [70-47-3]
fate and 20 mL of sulfuric acid. Heat the mixture at a low
Colorless crystals. One g dissolves in 50 mL of water; soluble
temperature until frothing ceases, then raise the tempera-
in acids and in alkalies; insoluble in alcohol and in ether. Its
ture and boil until the mixture acquires a pale yellow color
neutral or alkaline solutions are levorotatory; its acid solu-
or becomes practically colorless. Cool the flask, add about
tions are dextrorotatory.
250 mL of water, and cautiously add sodium hydroxide so-
Specific rotation 781: between +31 and +33, deter-
lution (3 in 10) until the contents are alkaline, then add
mined in a solution in diluted hydrochloric acid containing
5 mL more. Connect the flask at once by means of a spray
the equivalent of 5 g (on the anhydrous basis, as deter-
trap to a condenser, the lower outlet tube of which dips
mined by drying at 105 for 5 hours) in each 100 mL.
beneath the surface of 50.0 mL of 0.1 N sulfuric acid VS
contained in a receiving flask. Distill the mixture until about
Chloride (Reagent test)One g shows not more than 100 mL of distillate has been collected in the acid. Add
0.03 mg of Cl (0.003%). methyl red TS, and titrate the excess acid with 0.1 N so-
dium hydroxide VS. Each mL of 0.1 N sulfuric acid is equiva- Sulfate (Reagent test, Method I )One g shows not more
lent to 1.401 mg of N. Not less than 60 mg of nitrogen is than 0.05 mg of SO4 (0.005%).
found.
Heavy metals (Reagent test): 0.002%.
Assay for nitrogen as ammoniaTo 100 mL of test solu-
Nitrogen content, Method II 461: between 18.4% and
tion, contained in a 500-mL Kjeldahl flask, add 5 g of bar-
18.8% of N is found.
ium carbonate and 100 mL of water, and by means of a
L-Aspartic Acid, C4H7NO4133.1 [56-84-8]White to
spray trap, connect the flask to a condenser, the lower out-
off-white powder. Use a suitable grade.
let tube of which dips beneath the surface of 50.0 mL of 0.1
Azure A, C14H14ClN3S291.80 [531-53-3]Use a suit-
N sulfuric acid VS contained in a receiving flask. Distill the
able grade.
mixture until about 100 mL of distillate has been collected,
Bacterial Alkaline Protease PreparationUse a suitable
add methyl red TS, and titrate the excess acid with 0.1 N
grade.
sodium hydroxide VS. Each mL of 0.1 N sulfuric acid is
[NOTEA suitable grade is commercially available as
equivalent to 1.703 mg of NH3. The amount of ammonia
Protex 6L from Genencor, www.genencor.com, or as
found does not exceed 0.35% of the total solids in the por-
Optimase Enzyme from Solvay Enzymes Inc., www.
tion of test solution taken.
solvaypharmaceuticals.com.]
Total solidsDistribute 10 mL of test solution over clean, Barbital Sodium, C8H11N2NaO3206.2 [144-02-5]
dry sand or asbestos, tared in a porcelain dish, and dry at White, crystalline powder or colorless crystals. Freely soluble
105 for 16 hours: the residue weighs not less than 750 mg in water; slightly soluble in alcohol. Use a suitable reagent
(75%). grade.
Barbituric Acid, C4H4N2O3128.09 [65-52-7] Residue on ignitionIncinerate the residue obtained in
Faintly beige powder. Soluble in water, in alcohol, in chloro- the test for Total solids by heating the dish to a dull-red
form, and in ether. Melts with decomposition at 251.6. Use heat: the residue does not exceed 30% of the total solids.
a suitable grade.
Chlorides calculated as sodium chlorideDissolve the ash
Barium Acetate, C4H6BaO4255.43 [543-80-6]Use
obtained in the test for Residue on ignition in about 50 mL of
ACS reagent grade.
water, and carefully transfer to a 100-mL volumetric flask.
Barium Chloride, BaCl2 2H2O244.26
Add to the solution a few drops of nitric acid and 10.0 mL
of 0.1 N silver nitrate VS. Add water to volume, and mix.
Barium Chloride, Anhydrous, BaCl2208.23
Filter into a dry flask through a dry filter, rejecting the first
[10361-37-2]This may be made by drying barium chloride
10 mL of the filtrate. To 50.0 mL of the subsequent filtrate
in thin layers at 125 until the loss in weight between two
add 1 mL of ferric ammonium sulfate TS, and titrate with
successive, 3-hour drying periods does not exceed 1%.
0.1 N ammonium thiocyanate VS. Each mL of 0.1 N silver
nitrate is equivalent to 5.844 mg of NaCl. The weight of
chlorides calculated as sodium chloride obtained, when mul-
tiplied by 2, does not exceed 6% of the total solids.
Alcohol-insoluble substancesTransfer 25 mL of test solu-
tion to a 100-mL conical flask, add 50 mL of alcohol, and
shake thoroughly. Collect the precipitate on a counterpoised
filter, wash it three times with a mixture of 2 volumes of
alcohol and 1 volume of water, and dry at 105 for 2 hours: and Technology, Office of Standard Reference Materials,
the weight of the precipitate, representing the alcohol-insol- www.nist.gov, as standard sample No. 350.]
uble solids, does not exceed 10% of the total solids in the Benzophenone, (C6H5)2CO182.22 [119-61-9]
portion of test solutiontaken. White, crystalline powder.
NitrateBoil 10 mL of test solution for 1 minute with Melting range 741: between 47 and 49.
1.5 g of activated charcoal, add water to replace that lost by p-Benzoquinone (Quinone), C6H4O2108.09
evaporation, filter, and add 1 drop of the filtrate to 3 drops [106-51-4]Dark yellow powder having a green cast.
of a 1 in 100 solution of diphenylamine in sulfuric acid: no Slightly soluble in water; soluble in alcohol, in ether, and in
blue color is produced. fixed alkali solutions. May darken on standing. Darkened
Benzaldehyde, C7H6O106.12 [100-52-7]Color- material may be purified by sublimation in vacuum.
less, strongly refractive liquid. Soluble in water; miscible
with alcohol, with ether, and with fixed and volatile oils.
Benzoyl Chloride, C6H5COCl140.57 [98-88-4]Use
AssayPipet about 1 mL into a tared, glass-stoppered ACS reagent grade.
weighing bottle, and weigh accurately. Loosen the stopper, N-Benzoyl-L-arginine Ethyl Ester Hydrochloride,
and transfer both the weighing bottle and its contents to a C15H22N4O3 HCl342.82Determine suitability for use as a
250-mL conical flask containing 25 mL of a hydro-alcoholic substrate as directed under Crystallized Trypsin (USP mono-
solution of hydroxylamine hydrochloride (prepared by dis- graph).
solving 34.7 g of hydroxylamine hydrochloride in 160 mL of 3-Benzoylbenzoic Acid, C14H10O3226.23
water, then adding alcohol to make 1000 mL, and neutraliz- [579-18-0]White to off-white powder.
ing to bromophenol blue by the addition of sodium hydrox-
AssayPrepare a mixture of 1% trifluoroacetic acid in
ide TS). Using a graduated cylinder to measure the volume,
water and 1% trifluoroacetic acid in acetonitrile (55:45) for
rinse the sides of the flask with an additional 50 mL of this
the mobile phase. Inject about 20 L into a suitable liquid
reagent solution. Allow the solution to stand for 10 minutes,
add 1 mL of bromophenol blue TS, and titrate the liberated
a 230-nm detector and a 4.6-mm 15-cm column that
hydrochloric acid with 1 N sodium hydroxide VS. Perform a
contains packing L1. The flow rate is about 1.5 mL per min-
blank determination with the same quantities of the same
ute. The area of the C14H10O3 peak is not less than 98.5% of
reagents, and make any necessary correction. Each mL of
the total peak area.
1 N sodium hydroxide consumed is equivalent to 106.1 mg
Benzoylformic Acid (Phenylglyoxylic Acid),
of C7H6O. Not less than 98% is found.
C6H5COCO2H150.14 [611-73-4]Powder. Soluble in
Specific gravity 841: between 1.041 and 1.046. methanol.
Refractive index 831: between 1.5440 and 1.5465 at Melting range 741: between 62 and 67.
20. Benzphetamine Hydrochloride, C17H21N HCl275.82
[5411-22-3]White to off-white, crystalline powder. Freely
Hydrocyanic acidShake 0.5 mL with 5 mL of water, add
soluble in water, in alcohol, and in chloroform; slightly solu-
0.5 mL of sodium hydroxide TS and 0.1 mL of ferrous sul-
ble in ether.
fate TS, and warm the mixture gently. Add a slight excess of
hydrochloric acid: no greenish-blue color or blue precipitate AssayDissolve about 500 mg, accurately weighed, in a
is observed within 15 minutes. mixture of 50 mL of glacial acetic acid and 10 mL of mercu-
Benzalkonium ChlorideUse Benzalkonium Chloride (NF ric acetate TS, add 1 drop of crystal violet TS, and titrate
monograph). with 0.1 N perchloric acid VS to a blue-green endpoint.
Benzamidine Hydrochloride Hydrate (Benzenecarbox- Perform a blank determination, and make any necessary
imidamide Monohydrochloride, Hydrate), C7H8N2 HCl correction. Each mL of 0.1 N perchloric acid is equivalent to
xH2O156.6 [206752-36-5]White to off-white powder. 27.58 mg of C17H21N HCl. Between 98.0% and 101.0%,
Use a suitable grade. calculated on the dried basis, is found.
[NOTEA suitable grade is available from Sigma-Aldrich,
www.sigma-aldrich.com.]
Benzanilide, C13H11NO197.23 [93-98-1]Off-white,
mined in a solution containing 200 mg in 10 mL, the speci-
light gray to grayish-green powder. Insoluble in water; spar-
men having been previously dried in vacuum at 60 for
ingly soluble in alcohol; slightly soluble in ether.
3 hours.
Loss on drying 731Dry it in vacuum at 60 for
Solubility in acetoneA 1.0-g portion dissolves completely
3 hours: it loses not more than 1% of its weight.
in 50 mL of acetone to yield a clear solution.
Residue on ignition 281: not more than 0.2%.
Benzene, C6H678.11 [71-43-2]Use ACS reagent
2-Benzylaminopyridine, C12H12N2184.24
grade.
Benzenesulfonamide, C6H5SO2NH2157.19
1-Benzylimidazole, C10H10N2158.20 [4238-71-5]
[98-10-2]White to pale beige crystals.
White crystals.
AssayTransfer about 40 mg, accurately weighed, to a
Benzenesulfonyl Chloride, C6H5SO2Cl176.62
100-mL beaker. Dissolve in 50 mL of glacial acetic acid. Ti-
[98-09-9]Colorless, oily liquid. Insoluble in cold water; sol-
trate with 0.1 N perchloric acid VS, determining the
uble in alcohol and in ether. Solidifies at 0.
endpoint potentiometrically using a combination calomel-
platinum electrode. Perform a blank determination and
Boiling range (Reagent test): between 251 and 252. make any necessary correction. Each mL of 0.1 N perchloric
Benzhydrol (-Phenylbenzenemethanol), C13H12O184.23 acid is equivalent to 15.82 mg of C10H10N2. Not less than
[91-01-0]White to pale yellow crystals. Very slightly solu- 99% is found.
ble in water; soluble in alcohol, in ether, and in chloroform. Benzyltrimethylammonium Chloride,
C6H5CH2N(CH3)3Cl185.69 [56-93-9]Available as a
60% aqueous solution. Is clear and is colorless or not more
than slightly yellow.
ceed 2.
Benzoic Acid, C6H5COOH122.12 [65-85-0]Use AssayPipet 2 mL into a 50-mL volumetric flask, and add
ACS reagent grade. water to volume. Pipet 20 mL of the solution into a 125-mL
[NOTEBenzoic Acid of a quality suitable as a primary conical flask, add about 30 mL of water, then add 0.25 mL
standard is available from the National Institute of Standards of dichlorofluorescein TS, and titrate with 0.1 N silver nitrate
VS. Each mL of 0.1 N silver nitrate is equivalent to 18.57 mg Biphenyl, C12H10154.21 [92-52-4]Colorless to
of C6H5CH2N(CH3)3Cl. Between 59.5% and 60.5% is found. white crystals or crystalline powder. Insoluble in water; solu-
Beta-lactamaseBeta-lactamase is an enzyme produced ble in alcohol and in ether. Boils at about 254.
by a variety of bacteria, but is usually obtained from culture
filtrates of a strain of Bacillus cereus. It has the specific prop-
2,2-Bipyridine (,-Dipyridyl), C10H8N2156.18
erty of inactivating penicillins and cephalosporins by split-
[366-18-7]White or pink, crystalline powder. Soluble in
ting the bond linking the nitrogen of the thiazolidine to the
water and in alcohol. Melts at about 69, and boils at about
adjacent carbonyl carbon.
272.
It occurs in the form of small, brown, easily pulverizable
SensitivenessPrepare the following solutions: (A)Dis-
pieces or granules. Freely soluble in water, forming a slightly
solve 350 mg of ferrous ammonium sulfate in 50 mL of
opalescent solution that is practically neutral to litmus pa-
water containing 1 mL of sulfuric acid, and add 500 mg of
per. Is precipitated from its water solutions by acetone, by
hydrazine sulfate, then add water to make 500 mL. For use,
alcohol, and by dioxane, and is inactivated by contact with
dilute this solution with water in the ratio of 1 in 100 mL.
these solvents. Is rapidly inactivated by ethyl acetate and is
(B)Dissolve 8.3 g of sodium acetate and 12 mL of glacial
irreversibly destroyed at a temperature of about 80.
acetic acid in water to make 100 mL. Add 1 mL of a solution
Beta-lactamase is assayed by a procedure depending upon
of the specimen (1 in 1000) to a mixture of 10 mL of water
a determination of the amount of penicillin G potassium or
and 1 mL of each of solutions A and B: a pink color results
penicillin G sodium destroyed at a pH of 7.0 in a solution of
immediately.
such concentration that the inactivation proceeds as a zero-
SolubilityA 100-mg portion dissolves completely in
order reaction.
10 mL of water.
BetanaphtholSee 2-Naphthol.
Residue on ignition (Reagent test): not more than 0.2%. Bibenzyl (Dibenzyl), C14H14182.26 [103-29-7]Col-
4,4-Bis(4-amino-1-naphthylazo)-2,2-stilbenedisulfonic orless crystals. Freely soluble in chloroform and in ether;
Acid, C34H26N6O6S2678.74 [5463-64-9]Use a suitable sparingly soluble in alcohol; practically insoluble in water.
grade.
[NOTEA suitable grade is available from TCI America,
Bile SaltsA concentrate of beef bile, the principal con-
www.tciamerica.com.]
stituent of which is sodium desoxycholate, determined as
Bis(2-ethylhexyl) Maleate, C20H36O4340.50
cholic acid. Soluble in water and in alcohol; the solutions
[142-16-5]Colorless to pale yellow, clear liquid. Miscible
foam strongly when shaken.
with acetone and with alcohol. Specific gravity about 0.945.
Insoluble substancesDissolve 5 g in 100 mL of dilute al-
AssayPlace about 2.5 g, accurately weighed, in a
cohol (84 in 100), warming if necessary to aid solution. Fil-
250-mL flask, add 50.0 mL of 0.5 N alcoholic potassium hy-
ter within 15 minutes through a tared filter, and wash with
droxide VS, and reflux for 45 minutes. Cool, add 0.5 mL of
small portions of the dilute alcohol until the last washing is
phenolphthalein TS, and titrate the excess alkali with 0.5 N
colorless or practically so, then dry the residue at 105 for
hydrochloric acid VS. Perform a blank determination at the
1 hour, and weigh: the weight of the residue does not ex-
same time, using the same amount of 0.5 N alcoholic po-
ceed 0.1%.
tassium hydroxide (see Residual Titrations under Titrimetry
Assay
541). The difference, in mL, between the volumes of 0.5 N
STANDARD CHOLIC ACID SOLUTIONDissolve 50.0 mg of cholic
hydrochloric acid consumed in the test titration and blank
acid, accurately weighed, in dilute acetic acid (6 in 10) to
titration, multiplied by 85.1, represents the quantity, in mg,
make 100 mL, and mix. Store in a refrigerator.
of bis(2-ethylhexyl) maleate in the portion taken. Not less
PROCEDUREDissolve 1.0 g, accurately weighed, in 50 mL than 97% is found.
of dilute acetic acid (6 in 10). Filter the solution, if neces- Bis(2-ethylhexyl) Phthalate, C6H4-1,2-
sary, into a 100-mL volumetric flask, wash the original con- [COOCH2(C2H5)CH(CH2)3CH3]2390.56 [117-81-7]Col-
tainer and the filter with small portions of dilute acetic acid orless to light yellow liquid.
(6 in 10), add the same acetic acid to volume, and mix.
Refractive index 831: between 1.4855 and 1.4875, at
Dilute 10 mL of this solution, accurately measured, with di-
20.
lute acetic acid (6 in 10) to make 100 mL, and mix.
Bis(2-ethylhexyl) Sebacate (Dioctyl Sebacate),
Pipet 1 mL each of the Standard Cholic Acid Solution and
C8H17OOC(CH2)8COOC8H17426.67 [122-62-3]Pale
the solution of the Bile Salts into two matched test tubes.
straw-colored liquid. Insoluble in water. Refractive index
To each tube add 1 mL, accurately measured, of freshly pre-
about 1.448. Suitable for use in gas chromatography.
pared furfural solution (1 in 100), immediately place the
Specific gravity, 20/20841: between 0.913 and
tubes in an ice-bath for 5 minutes, then add to each tube
0.917.
13 mL, accurately measured, of dilute sulfuric acid, made by
Boiling range: between 243 and 248 at 5 mm of mer-
cautiously mixing 50 mL of sulfuric acid with 65 mL of
cury.
water. Mix the contents of the tubes, and place them in a
[NOTEA suitable grade is Dioctyl Sebacate, available
water bath maintained at a temperature of 70 for 10 min-
utes. Immediately transfer the tubes to an ice-bath for
Bis(2-ethylhexyl)phosphoric Acid [Bis(2-ethylhex-
2 minutes, then determine the absorbance of each solution
yl)Phosphate], [CH3(CH2)3CH(C2H5)CH2]2HPO4322.42
at the wavelength of maximum absorbance at about 670
[298-07-7]Light yellow, viscous liquid. Insoluble in water;
nm, with a suitable spectrophotometer. Calculate the quan-
freely soluble in chloroform and in ethyl acetate. Refractive
tity, in mg, of cholic acid (C24H40O5) in the weight of the
index: about 1.443. Specific gravity: about 0.997.
Bile Salts taken by the formula:
AssayDissolve about 250 mg, accurately weighed, in
500(AU/AS) 50 mL of dimethylformamide, add 3 drops of a 1 in 100
solution of thymol blue TS in dimethylformamide, and ti-
in which AU and AS are the absorbances of the solutions trate with 0.1 N sodium methoxide VS to a blue endpoint.
from the Bile Salts and the Standard Cholic Acid Solution, Perform a blank determination, and make any necessary
respectively. Not less than 45% of cholic acid is found. correction. Each mL of 0.1 N sodium methoxide is equiva-
lent to 32.24 mg of (C8H17)2HPO4. Between 95% and 105%
is found.
SolubilityOne volume dissolves in 9 volumes of chloro- [NOTEThere are many manufacturers and suppliers of
form to yield a clear solution, and 1 volume dissolves in 9 these reagents that are licensed by the Center for Biologics
volumes of ethyl acetate to yield a clear solution. Evaluation and Research Food and Drug Administration.
ColorA 1 in 100 solution in chloroform exhibits an ab-
sorptivity of not more than 0.03 at 420 nm.
Bis(4-sulfobutyl) Ether Disodium, Na2C8H16O7S2
Anti-A Blood Grouping Reagent, Anti-B Blood
334.32Use a suitable grade with a content of NLT 95%.
Grouping Reagent, and Anti-AB Blood Grouping
ReagentThe reagents can be monoclonal or polyclonal
RM-969-C50 from www.cydexpharma.com.]
and must be obtained from manufacturers or suppliers li-
Bismuth Nitrate Pentahydrate, Bi(NO3)3 5H2O485.07
censed by the Center for Biologics Evaluation and Research,
Food and Drug Administration for use in microplate tests.
Bismuth Subnitrate (Bismuth(III) Nitrate Basic),
The use of reagents from an unlicensed manufacturer or
Bi5O (OH)9(NO3)41461.99 [1304-85-4]Use Bismuth
supplier may invalidate the results. Generally, all three re-
Subnitrate (USP monograph).
agents are available as part of a kit.
Bismuth Sulfite AgarUse a suitable grade.
Bis(trimethylsilyl)acetamide (N,O-Bis(trimethylsil-
yl)acetamide; BSA), CH3CON[Si(CH3)3]2203.43
[10416-59-8]Clear, colorless liquid. Readily hydrolyzes
when exposed to moist air. Handle under nitrogen, and
store in a cool place.
AssayNot less than 90% of CH3CON[Si(CH3)3]2, a suita-
Blue Tetrazolium (3,3-(3,3-Dimethoxy[1,1-biphenyl]-4,4-
ble gas chromatograph equipped with a thermal conductiv-
diyl)bis[2,5-diphenyl-2H-tetrazolium]dichloride),
ity detector being used. The following conditions are suita-
C40H32Cl2N8O2727.64 [1871-22-3]Lemon-yellow crys-
ble and provide a retention time of approximately
tals. Slightly soluble in water; freely soluble in chloroform
15 minutes.
and in methanol; insoluble in acetone and in ether.
COLUMN: 3-mm 1.83-m stainless steel containing 5%
Solubility in methanolDissolve 1 g in 100 mL of metha-
phase G1 on support S1A.
nol: complete solution results, and the solution is clear.
INJECTION TEMPERATURE: 160.
ColorTransfer a portion of the methanol solution ob-
COLUMN TEMPERATURE: 90, programmed to rise 4 per min-
tained in the preceding test to a 1-cm cell, and determine
ute to 160.
its absorbance at 525 nm, against water as the blank: the
CARRIER GAS: Helium. absorbance does not exceed 0.20.
Refractive index 831: between 1.4150 and 1.4170 at Molar absorptivity 851Its molar absorptivity in metha-
20. nol, at 252 nm, is not less than 50,000.
Bis(trimethylsilyl)trifluoroacetamide (N,O-Bis(trimethyl-
Suitability test
silyl)trifluoroacetamide; BSTFA), CF3CON[Si(CH3)3]2257.40
STANDARD PREPARATIONDissolve in alcohol a suitable quan-
[25561-30-2]Clear, colorless liquid. Readily hydrolyzes
tity of USP Hydrocortisone RS, previously dried at 105 for
when exposed to moist air. Store in a cool place.
3 hours and accurately weighed, and prepare by stepwise
AssayNot less than 98% of CF3CON[Si(CH3)3]2, a suita-
dilution a solution containing about 10 g per mL.
ble gas chromatograph equipped with a thermal conductiv-
PROCEDUREPipet 10-, 15-, and 20-mL portions of Stan-
ity detector being used. The following conditions are suita-
dard Preparation into separate, glass-stoppered, 50-mL coni-
ble and provide a retention time of approximately
cal flasks. Add 10 mL and 5 mL, respectively, of alcohol to
15 minutes.
the flasks containing the 10- and 15-mL portions of Stan-
COLUMN: 3-mm 1.83-m stainless steel containing 5%
dard Preparation, and swirl to mix. To each of the flasks, and
phase G1 on support S1A.
to a fourth flask containing 20 mL of alcohol, add 2.0 mL of
INJECTION TEMPERATURE: 170. a solution prepared by dissolving 50 mg of blue tetrazolium
in 10 mL of alcohol, mix, and then add 2.0 mL of a solution COLUMN TEMPERATURE: 70, programmed to rise 4 per min-
prepared by diluting 1 mL of tetramethylammonium hy- ute to 140.
droxide TS with alcohol to 10 mL. Mix, allow the flasks to
CARRIER GAS: Helium.
stand in the dark for 90 minutes, and determine the ab-
sorbances of the three solutions of the steroid standard at
20.
525 nm, with a suitable spectrophotometer, using the solu-
Bis(trimethylsilyl)trifluoroacetamide with Trimethyl-
tion in the fourth flask as the blank. Plot the absorbances on
chlorosilane [25561-30-2]Use a suitable grade.
the abscissa and the amount of hydrocortisone on the ordi-
nate scale of arithmetic coordinate paper, and draw the
curve of best fit: the absorbance of each solution is propor-
Blood (for carbon monoxide test in gases)Use oxalated
tional to the concentration, and the absorbance of the solu-
or defibrinated blood of dogs, sheep, cattle, or human beings
tion containing 200 g of hydrocortisone is not less than
within 24 hours after bleeding. Prepare oxalated blood by
0.50.
adding 10 mg of sodium oxalate to each mL of the freshly
Boric Acid, H3BO361.83 [10043-35-3]Use ACS rea-
drawn blood.
gent grade.
Blood Group A1 Red Blood Cells and Blood Group B
()-Bornyl Acetate (1,7,7-Trimethylbicyclo[2,2,1]-heptan-
Red Blood CellsThese cells must be obtained from manu-
2-ol acetate), C12H20O2196.29 [5655-61-8]Use a
facturers or suppliers licensed by the Center for Biologics
suitable grade.
Evaluation and Research/Food and Drug Administration. The
Boron Trifluoride, BF367.81 [7637-07-2]Use a
use of reagents from an unlicensed manufacturer or supplier
suitable grade.
may invalidate the results. Generally, they are available as
14% Boron TrifluorideMethanol [373-57-9]Use a
part of a kit for ABO Blood Group testing. The cells licensed
suitable grade.
for use in test tubes can also be used in the microtiter plate
Bovine CollagenUse a suitable grade that contains less
method described in the monographs of Red Blood Cells and
than 1 g glycosaminoglycan per mg.
Whole Blood.
[NOTEA suitable grade is available from Sigma-Aldrich, in water. Miscible with alcohol and with ether. Boils at
www.sigma-aldrich.com.] about 88.
7 Percent Bovine Serum Albumin Certified Standard
Assay
Available from the National Institute of Standards and Tech-
HYDROXYLAMINE HYDROCHLORIDE SOLUTIONDissolve 20 g of
nology, www.nist.gov, as SRM 927.
hydroxylamine hydrochloride in 40 mL of water, and dilute
Branched Polymeric Sucroseabout 400 kDa
with alcohol to 400 mL. Add, with stirring, 300 mL of 0.5 N
[26873-85-8]White to off-white powder. Synthetic poly-
alcoholic potassium hydroxide VS, and filter. Discard after 2
mer made by the copolymerization of sucrose and
days.
epicholrohydrin. Use a suitable grade.
PROCEDURETransfer about 1 g, accurately weighed, to a
[NOTEA suitable grade is available commercially as
glass-stoppered, 250-mL flask, add 75.0 mL of Hydroxyla-
Ficoll from Pharmacia Fine Chemicals, Inc., 800 Centen-
mine hydrochloride solution,and insert the stopper in the
nial Ave., Piscataway, NJ 08854.]
flask. Reflux the mixture for 1 hour, then cool to room tem-
Brilliant Green (Malachite Green G), C27H34N2O4S
perature. Add bromophenol blue TS, and titrate with 0.5 N
482.64 [3051-11-4]Glistening, golden-yellow crystals.
hydrochloric acid VS to a greenish-yellow endpoint. [NOTE
Soluble in water and in alcohol. Absorption maximum: 623
Alternatively, the solution may be titrated potentiometrically
nm.
to a pH of 3.4.] Perform a blank test with the same quanti-
BromelainA proteolytic enzyme isolated from pineap-
ties of reagent used for the test specimen, and make any
ple. Use a suitable grade.
necessary correction. Each mL of 0.5 N hydrochloric acid is
Bromine, BrAt. Wt. 79.904 [7726-95-6]Use ACS
equivalent to 43.05 mg of CH3COCOCH3. Not less than
reagent grade.
97% of CH3COCOCH3 is found.
-Bromo-2-acetonaphthone (Bromomethyl 2-naphthyl
Congealing temperature 651: between 2.0 and ketone), C12H9BrO249.10Tannishpink crystals.
5.5.
Refractive index 831: between 1.3935 and 1.3965, at p-Bromoaniline, C6H6BrN172.02 [106-40-1]White
20. to off-white crystals. Insoluble in water; soluble in alcohol
and in ether. Specific gravity 841: about 0.98.
1-Butanesulfonic Acid Sodium Salt (Sodium AssayTransfer about 650 mg, accurately weighed, to a
1-butanesulfonate), C4H9NaO3S160.16 [2386-54-1] suitable container, and dissolve in 50 mL of glacial acetic
Use a suitable grade with a content of not less than 99.0%. acid TS. Add crystal violet TS, and titrate with 0.1 N per-
1,4-Butane Sultone (4-Hydroxybutane-1-sulfonic Acid chloric acid VS. Perform a blank determination, and make
delta-sultone), C4H8O3S136.17 [1633-83-6]Use a suit- any necessary correction. Each mL of 0.1 N perchloric acid
able grade with a content of NLT 99%. is equivalent to 17.20 mg of C6H6BrN. Not less than 98% is
ButanolSee Butyl Alcohol. found.
Butyl Acetate, Normal, CH3COO(CH2)3CH3116.16
Melting range 741: between 60 and 65, within a 2
range.
Butyl Alcohol (1-Butanol; Normal Butyl Alcohol), CH3
BromofluoromethaneUse a suitable grade.
(CH2)2CH2OH74.12 [71-36-3]Use ACS reagent grade.
N-Bromosuccinimide, C4H4BrNO2177.98
Butyl Alcohol, NormalSee Butyl Alcohol.
[128-08-5]White to off-white crystals or powder. Freely
Butyl Alcohol, Secondary (2-Butanol),
soluble in water, in acetone, and in glacial acetic acid.
CH3CH2CH(OH)CH374.12 [78-92-2]Use a suitable
[CAUTIONHighly irritating to eyes, skin, and mucous
grade with a content of NLT 99%.
membranes.]
Butyl Alcohol, Tertiary, (CH3)3COH74.12
AssayTransfer 200 mg, accurately weighed, to a conical
[75-65-0]Use ACS reagent grade tert-Butyl Alcohol.
flask, add 25 mL of 0.5 N alcoholic potassium hydroxide,
Butyl Benzoate, C11H14O2178.23 [136-60-7]Thick,
cover with a watch glass, heat to boiling, and boil for
oily, colorless to pale yellow liquid. Practically insoluble in
5 minutes. Cool, transfer the solution to a beaker, rinsing
water; soluble in alcohol and in ether.
the flask with water until the total volume of solution plus
AssayWhen examined by gas-liquid chromatography, it
rinsings is about 100 mL, and add 10 mL of glacial acetic
shows a purity of not less than 98%. The following condi-
acid. Insert suitable electrodes, and titrate with 0.1 N silver
tions have been found suitable for assaying it: use a 3-mm
nitrate VS, determining the endpoint potentiometrically.
1.8-m stainless steel column packed with liquid phase G4
Each mL of 0.1 N silver nitrate is equivalent to 17.80 mg of
on support S1A. Helium is the carrier gas, the injection port
C4H4BrNO2. Not less than 98% is found.
temperature is maintained at 180, the column temperature
Brucine Sulfate, (C23H26N2O4)2 H2SO4 7H2O1013.11
is maintained at 190, and the flame-ionization detector is
maintained at 280. The retention time is about 15 minutes.
BuffersSee Buffer Solutions under Solutions.
Refractive index 831: between 1.4980 and 1.5000, at 1,3-Butanediol (1,3-Butylene Glycol), C4H10O290.12
20. [107-88-0]Viscous, colorless liquid. Very hygroscopic. Solu-
n-Butyl Chloride (1-Chlorobutane), C4H9Cl92.57 ble in water, in alcohol, in acetone, and in methyl ethyl
[109-69-3]Clear, colorless, volatile liquid. [CAUTION ketone; practically insoluble in aliphatic hydrocarbons, in
Highly flammable.] Practically insoluble in water. Miscible benzene, and in toluene.
with alcohol and with ether. Use HPLC grade.
Butyl Ether (n-Dibutyl Ether), C8H18O130.23
Butyl Methacrylate, C8H14O2142.20 [97-88-1]Use
a suitable grade.
3-mm 1.8-m stainless steel column containing 20% phase
tert-Butyl Methyl Ether, C5H12O88.15
G16 on support S1A; the injection port temperature is
[1634-04-4]Colorless liquid.
maintained at 265; the column temperature is maintained
AssayInject an appropriate specimen into a gas chro- at 150 and programmed to rise 8 per minute to 210. The
matograph (see Chromatography 621) equipped with a area of the butanediol peak is not less than 98% of the total
flame-ionization detector, helium being used as the carrier peak area.
20.
2,3-Butanedione (Diacetyl), CH3COCOCH386.09
[431-03-8]Bright yellow to yellowish-green liquid. Soluble
the column temperature is maintained at ambient tempera-
ture and programmed to rise 10 per minute to 150. The AssayWeigh accurately about 500 mg, transfer to a
area of the C5H12O peak is not less than 99.8% of the total suitable container, add 30 mL of water, and mix. Add 40 mL
peak area. of water, and mix. Add phenolphthalein TS, and titrate with
0.1 N sodium hydroxide VS. Each mL of 0.1 N sodium hy-
droxide is equivalent to 8.81 mg of C4H8O2: not less than
n-Butylamine, CH3CH2CH2CH2NH273.14
99.0% of C4H8O2 is found.
[109-73-9]Colorless to pale yellow, flammable liquid. Mis-
cible with water, with alcohol, and with ether. Store it in Refractive index 831: about 1.398 at 20.
tight containers. Specific gravity: about 0.740. Butyrolactone (Dihydro-2-(3H)-furanone, -butyrolactone)
86.1 [96-48-0]Clear, colorless to practically colorless,
Distilling range, Method I 721Not less than 95% dis-
oily liquid. Miscible with water. Soluble in methanol and in
tills between 76 and 78.
ether.
Water, Method I 921: not more than 1.0%, determined
Boiling range 721: between 193 and 208.
by the Titrimetric Method.
Refractive index 831: about 1.435, at 20.
Chloride (Reagent test)One g (1.5 mL) shows not more
than 0.01 mg of Cl (0.001%). Specific gravity 841: between 1.128 and 1.135.
Butyrophenone (Phenyl Propyl Ketone), C10H12O148.21
Acidic impuritiesTo 50 mL add 5 drops of a saturated
[495-40-9]Use a suitable grade with a content of not less
solution of azo violet in benzene, and titrate quickly with
than 98.0%.
0.1 N sodium methoxide VS to a deep blue endpoint, ob-
Cadmium Acetate, C4H6CdO4 2H2O266.53
serving precautions to prevent absorption of atmospheric
[543-90-8]Colorless, transparent to translucent crystals.
carbon dioxide as by use of an atmosphere of nitrogen: not
Freely soluble in water; soluble in alcohol.
more than 1.0 mL of 0.1 N sodium methoxide is required
for neutralization. Insoluble matter (Reagent test): not more than 1 mg,
tert-Butylamine, C3H9CNH273.14 [75-64-9]Liquid. from 20 g (0.005%).
AssayInject an appropriate volume into a gas chromat- Chloride (Reagent test)One g shows not more than
ograph (see Chromatography 621) equipped with a flame- 0.01 mg of Cl (0.001%).
ionization detector, helium being used as the carrier gas.
Sulfate (Reagent test, Method II)Dissolve 10 g in
100 mL of water, add 1 mL of hydrochloric acid, and filter:
the residue weighs not more than 1.2 mg more than the
residue obtained in a complete blank test (0.005%).
230; the detector temperature is maintained at 300; the
Substances not precipitated by hydrogen sulfideDissolve
column temperature is maintained at 130 and pro-
2 g in a mixture of 135 mL of water and 15 mL of 1 N sulfu-
ric acid, heat to boiling, and pass a rapid stream of hydro-
C3H9CNH2 peak is not less than 99.5% of the total peak
gen sulfide through the solution as it cools. Filter, and to
area.
75 mL of the clear filtrate add 0.25 mL of sulfuric acid, then
evaporate to dryness, and ignite gently: the residue weighs
20.
not more than 1 mg (0.1%).
4-(Butylamino)benzoic Acid, C11H5NO2193.25
Cadmium Nitrate, Cd(NO3)2 4H2O308.48
[10325-94-7]Colorless, hygroscopic crystals. Very soluble
in water; soluble in alcohol.
Inc., P.O. Box 2060, Milwaukee, WI 53201; www.sigma-
aldrich.com.]
from 20 g (0.005%).
n-Butylboronic Acid (1-Butaneboronic Acid),
Chloride (Cl) (Reagent test)One g shows not more than
C4H9B(OH)2101.94 [4426-47-5]Use a suitable grade.
0.01 mg of Cl (0.001%).
[NOTEThis reagent is usually shipped and stored under
Sulfate (Reagent test, Method II)Evaporate a mixture of water. Before use, remove any excess water by light vacuum
12 g of specimen and 25 mL of hydrochloric acid on a filtration. A suitable grade is available from Sigma-Aldrich,
steam bath to dryness. Add another 15 mL of hydrochloric www.sigma-aldrich.com.]
acid, and again evaporate to dryness. Dissolve the residue in tert-Butyldimethylchlorosilane in N-Methyl-N-tert-
100 mL of water, filter, and add 1 mL of hydrochloric acid: butyldimethylsilyltrifluoroacetamide, (1 in 100)Use a
the residue weighs not more than 1.0 mg more than the suitable grade.
residue obtained in a blank test (0.003%). [NOTEA suitable grade is available as 99% MTBSTFA, 1%
TBDMCS from Regis Chemical Company, 8210 Austin Ave., Copper (Cu)Dissolve 0.5 g in 10 mL of water, add
P.O. Box 519, Morton Grove, IL 60053.] 10 mL of Ammonium Citrate Solution (see Lead 251), and
4-tert-Butylphenol, C10H14O150.22 [98-54-4] adjust the reaction to a pH of about 9 by the addition of
White, crystalline flakes or needles. Practically insoluble in 1 N ammonium hydroxide (about 30 mL). Add 1 mL of so-
water; soluble in alcohol and in ether. dium diethyldithiocarbamate solution (1 in 1000), and mix.
Add 5 mL of amyl alcohol, shake for about 1 minute, and Melting range 741: between 98 and 101.
allow the layers to separate: any yellow color in the amyl t-Butylthiol (tert-Butylthiol; tert-Butyl Mercaptan; 2-Methyl-
alcohol layer is not darker than that of a blank to which 2-propanethiol; TBM), (CH3)3CSH90.19 [75-66-1]Use a
0.01 mg of Cu has been added (0.002%). suitable grade with a content of NLT 98.0%.
[NOTEA suitable grade is available as catalog number Iron (Fe)Dissolve 1 g in 15 mL of water, add 2 mL of
20230 at www.sigma-aldrich.com.] hydrochloric acid, and boil for 2 minutes. Cool, and add
Butyraldehyde (Butanal), C4H8O72.11 [123-72-8] about 30 mg of ammonium persulfate and 15 mL of a solu-
Use a suitable grade, purified by redistillation, with a con- tion of potassium thiocyanate in normal butyl alcohol (made
tent of not less than 99.5%. by dissolving 10 g of potassium thiocyanate in 10 mL of
Butyric Acid, C4H8O288.11 [107-92-6]Clear, col- water, warming the solution to about 30, diluting with nor-
orless to faint yellow liquid. Miscible with water and with mal butyl alcohol to 100 mL, and shaking until clear). Shake
methanol. vigorously for 30 seconds, and allow the layers to separate:
any red color in the clear alcoholic layer is not darker than
that of a blank to which 0.01 mg of Fe has been added
(0.001%).
Lead (Pb)Dissolve 1.0 g in 10 mL of water, add 0.2 mL
of glacial acetic acid, and filter if necessary. To a 7-mL por-
tion of water add 0.2 mL of glacial acetic acid and 3 mL of
Standard Lead Solution (see Lead 251), and mix, to provide Calcium oxide and carbonateTriturate 1 g of calcium cit-
a blank. Then add to each solution 1.0 mL of potassium rate with 5 mL of water for 1 minute: the mixture does not
chromate solution (1 in 10), and mix: after 5 minutes, the turn red litmus blue. Then add 5 mL of warm 3 N hydro-
test solution is not more turbid than the blank (0.003%). chloric acid: only a few isolated bubbles escape.
Substances not precipitated by hydrogen sulfideDissolve Hydrochloric acid-insoluble matterDissolve 5 g by heating
2 g in 145 mL of water, add 5 mL of sulfuric acid (1 in 10), with a mixture of 10 mL of hydrochloric acid and 50 mL of
heat to boiling, and pass a rapid stream of hydrogen sulfide water for 30 minutes: not more than 2.5 mg of insoluble
through the solution as it cools. Filter, and to 75 mL of the residue remains (0.05%).
clear filtrate add 0.25 mL of sulfuric acid, then evaporate to
Loss on drying 731Dry it at 150 to constant weight:
dryness, and ignite gently: the residue weighs not more
it loses between 12.2% and 13.3% of its weight.
than 1 mg (0.1%).
Arsenic 211Proceed with 0.50 g as directed for or-
Calcium Acetate, Ca(C2H3O2)2 H2O176.18
ganic compounds (6 ppm of As).
[5743-26-0]White, crystalline granules or powder. Soluble
Heavy metals, Method I 231: 0.002%.
in about 3 parts of water; slightly soluble in alcohol. Use
Calcium Hydroxide [1305-62-0]Use ACS reagent grade.
ACS reagent grade.
Calcium Lactate, (CH3CHOHCOO)2Ca 5H2O308.29
Calcium Carbonate, CaCO3100.09 [471-34-1]Use
[814-80-2]White granules or powder. Is somewhat efflo-
ACS reagent grade.
rescent and at 120 becomes anhydrous. One g dissolves in
[NOTECalcium Carbonate of a quality suitable as a pri-
20 mL of water; practically insoluble in alcohol. Store it in
mary standard is available from the National Institute of
tight containers.
Standards and Technology, Office of Standard Reference
AssayAccurately weigh about 500 mg, previously dried Materials, www.nist.gov, as standard sample No. 915.]
at 120 for 4 hours, transfer to a suitable container, and Calcium Carbonate, Chelometric Standard, CaCO3
dissolve in 150 mL of water containing 2 mL of diluted hy- 100.09 [471-34-1]Use ACS reagent grade.
drochloric acid. Add 15 mL of sodium hydroxide TS and Calcium CaseinateWhite or slightly yellow, nearly
300 mg of hydroxy naphthol blue indicator, and titrate with odorless, powder. Insoluble in cold water, but forms a milky
0.05 M edetate disodium VS until the solution is deep blue. solution when suspended in water, stirred, and heated.
Each mL of 0.05 M edetate disodium is equivalent to
Residue on ignition (Reagent test)Ignite 5 g at 550: the
10.91 mg of C6H10CaO6. Not less than 98% is found.
residue weighs between 150 and 300 mg (3.0% to 6.0%).
Loss on drying 731Dry it at 120 for 4 hours: it loses
CalciumTreat the residue from the preceding test with
between 25.0% and 30.0% of its weight.
10 mL of diluted hydrochloric acid, filter, and to the clear
AcidityAdd phenolphthalein TS to 20 mL of a 1 in 20 filtrate add 5 mL of ammonium oxalate TS: it shows a white
solution, and titrate with 0.10 N sodium hydroxide: not precipitate upon standing.
more than 0.50 mL is required to produce a pink color.
Loss on drying 731Dry it in vacuum at 70 to constant
Heavy metals (Reagent test)Dissolve 1 g in 2.5 mL of weight: it loses not more than 7.0% of its weight.
diluted hydrochloric acid, dilute with water to 40 mL, and
FatSuspend 1.0 g in 5 mL of alcohol in a Mojonnier
add 10 mL of hydrogen sulfide TS: any brown color pro-
flask, add 0.8 mL of stronger ammonia water and 9 mL of
duced is not darker than that of a control containing
water, and shake. Add a second 5-mL portion of alcohol,
0.02 mg of added Pb (0.002%).
then add successive portions of 25 mL each of ether and
Magnesium and alkali saltsMix 1 g with 40 mL of water, solvent hexane, shaking after each addition by inverting the
carefully add 5 mL of hydrochloric acid, heat the solution, flask 30 times. Centrifuge, decant the solvent layer, evapo-
boil for 1 minute, and add rapidly 40 mL of oxalic acid TS. rate it at a low temperature, and dry on a steam bath: the
Add immediately to the warm mixture 2 drops of methyl residue weighs not more than 20 mg (2.0%).
red TS, then add ammonia TS dropwise, from a buret, until
Nitrogen content, Method I 461Between 12.5% and
the mixture is just alkaline. Cool to room temperature,
14.3% of N is found, calculated on the anhydrous basis.
transfer to a 100-mL graduated cylinder, dilute with water
Suspensibility in waterPlace 2 g in a beaker, and add
to 100 mL, mix, and allow to stand for 4 hours or overnight.
cool water slowly with stirring to form a thin, smooth paste.
Filter, and transfer to a platinum dish 50 mL of the clear
Add additional water to make a total of 100 mL. Stir, and
filtrate, to which has been added 0.5 mL of sulfuric acid.
heat to 80: a milky suspension is formed that does not
Evaporate the mixture on a steam bath to a small bulk.
settle after standing for 2 hours.
Carefully heat over a free flame to dryness, and continue
Calcium Chloride, CaCl2 2H2O147.01
heating to complete decomposition and volatilization of am-
[10035-04-8]Use ACS reagent grade Calcium Chloride Di-
monium salts. Finally ignite the residue at 800 25 for
hydrate.
15 minutes: the residue weighs not more than 5 mg (1%).
Calcium Chloride, Anhydrous (for drying), CaCl2
Volatile fatty acidStir about 500 mg with 1 mL of sulfu-
110.98 [10043-52-4]Use ACS reagent grade Calcium
ric acid, and warm: the mixture does not emit an odor of
Chloride Desiccant.
volatile fatty acid.
Calcium Citrate, Ca3(C6H5O7)2 4H2O570.49
Calcium Nitrate, Ca(NO3)2 4H2O236.15
[813-94-5]A white, crystalline powder. Slightly soluble in
water; freely soluble in 3 N hydrochloric acid and in 2 N
Calcium Pantothenate, DextroUse Calcium Pantothen-
nitric acid; insoluble in alcohol. To 15 mL of hot 2 N sulfuric
ate (USP monograph).
acid add in small portions and with stirring about 500 mg
Calcium Sulfate, CaSO4 2H2O172.17
of calcium citrate. Boil the mixture for 5 minutes, and filter
while hot: the cooled filtrate responds to the identification
Calconcarboxylic Acid (2-Naphthalenecarboxylic acid,
test for Citrate 191.
3-hydroxy-4-[(2-hydroxy-4-sulfo-1-naphthalenyl)azo]; Calcon-
AssayAccurately weigh about 400 mg of the salt, previ-
3-carboxylic Acid; Cal-Red), C21H14N2O7S438.42
ously dried at 150 to constant weight, and transfer to a
250-mL beaker. Dissolve the test specimen in 150 mL of
Calconcarboxylic Acid TriturateMix 1 part of calcon-
water containing 2 mL of 3 N hydrochloric acid, add 15 mL
carboxylic acid with 99 parts of sodium chloride.
of 1 N sodium hydroxide and 250 mg of hydroxy naphthol
Test for sensitivityDissolve 50 mg of calconcarboxylic
blue, and titrate with 0.05 M edetate disodium VS until the
acid triturate in a mixture of 2 mL 10 N sodium hydroxide
solution turns deep blue. Each mL of 0.05 M edetate diso-
and 100 mL of water. The solution is blue but becomes vio-
dium is equivalent to 8.307 mg of Ca3(C6H5O7)2: between
let on addition of 1 mL of a 10 g per L solution of magne-
97.5% and 101% is found.
sium sulfate and 0.1 mL of a 1.5 g per L solution of calcium
chloride and turns pure blue on addition of 0.15 mL of 0.01 Measuring range: 5 to 150 ppm.
M sodium edetate. [NOTEAvailable from Draeger Safety, Inc.,
Calf Thymus DNAUse a suitable grade. [NOTEA suita- www.draeger.com, or from Gastec Corp., www.gastec.co.jp,
ble grade is commercially available from Worthington Bio- distributed in the USA by www.nextteq.com.]
chemical Corp., www.worthington-biochem.com.] Carbon Tetrachloride, CCl4153.82 [56-23-5]Use
dl-Camphene, C10H16136.24 [79-92-5]Use a suita- a grade meeting the specifications of ACS Reagent Chemi-
ble grade. cals, 8th Edition.
[NOTEA suitable grade is available as camphene, 95%, Carboxylate (Sodium Form) Cation-exchange Resin
catalog number 45,606-5, from Sigma-Aldrich, www.sigma- (50- to 100-mesh)See Cation-Exchange Resin, Carboxylate
aldrich.com.] (Sodium Form) (50- to 100-mesh).
d-10-Camphorsulfonic Acid [(1S)-(+)-10-Camphorsulfonic Carboxymethoxylamine Hemihydrochloride,
acid; (1S)-Camphor-10-sulfonic acid; (+)-Camphor-10-sulfonic 2(C2H5NO3) HCl218.59 [2921-14-4]White, crystal-
acid ()], C10H16O4S232.30 [3144-16-9]Use a suitable line powder. Use a suitable grade.
grade. Carmine (Alum Lake of Carminic Acid), C22H20O13 xAl
[NOTEA suitable grade is available as catalog number [1390-65-4]Red powder. Use a suitable grade.
C2107 from www.sigma-aldrich.com.] (R)-()-Carvone (2-Methyl-5-(1-methylethenyl)-2-cyclohex-
dl-10-Camphorsulfonic Acid, [Camphor-10-sulfonic acid ene-1-one), C10H14O150.22 [6485-40-1]Use a suitable
()]; [(RS)-10-Camphorsulfonic acid], C10H16O4S232.30 grade.
[5872-08-2] Is optically inactive. Casein [9000-71-9]White or slightly yellow, granular
Use a suitable grade. powder. Insoluble in water and in other neutral solvents;
[NOTEA suitable grade is available as catalog number readily dissolved by ammonia TS and by solutions of alkali
147923 from www.sigma-aldrich.com.] hydroxides, usually forming a cloudy solution.
Canada Balsam [8007-47-4]A natural product derived
Residue on ignition (Reagent test)Ignite 2 g: the residue
from the resin of Abies balsamea. Use a suitable grade.
Canola Oil [120962-03-0]Use a suitable grade.
Capric Acid (Decanoic Acid), C10H20O2172.26
[334-48-5]White, solidified melt or fragments. Soluble in
AlkalinityShake 1 g with 20 mL of water for 10 minutes,
alcohol, in chloroform, and in ether; practically insoluble in
and filter: the filtrate is not alkaline to red litmus paper.
water.
Soluble substancesWhen the filtrate from the Alkalinity
AssayInject an appropriate sample dissolved in acetone
test is evaporated and dried at 105, the residue weighs not
into a gas chromatograph (see Chromatography 621) that
more than 1 mg (0.1%).
is equipped with a flame-ionization detector and contains a
FatsDissolve 1 g in a mixture of 10 mL of water and 0.53-mm 30-m capillary column coated with a layer of
5 mL of alcoholic ammonia TS, and shake out with two phase G25. The carrier gas is helium, flowing at a rate of
20-mL portions of solvent hexane. Evaporate the hexane at 9 mL per minute. The chromatograph is programmed as fol-
a low temperature, and dry at 80: the weight of the resi- lows. Initially the column temperature is equilibrated at
due does not exceed 5 mg (0.5%). 150, then the temperature is increased at a rate of 10 per
minute to 250. The injection port temperature is main- Nitrogen content, Method I 461: between 15.2% and
tained at 240, and the detector temperature at 265. The 16.0% of N is found, on the anhydrous basis.
area of the capric acid peak is not less than 98.5% of the Where vitamin-free casein is required, use casein that has
total peak area. been rendered free from the fat-soluble vitamins by continu-
ous extraction with hot alcohol for 48 hours followed by air- Melting range 741: between 30 and 33.
drying to remove the solvent. Carbazole, C12H9N167.21 [86-74-8]Off-white to
Casein, Hammersten [9000-71-9]. [NOTEA suitable tan powder.
grade is available from www.emdchemicals.com, catalog
AssayInject an appropriate volume into a gas chromat-
number CX0525-1.]
ograph (see Chromatography 621) equipped with a flame-
Catechol (o-Dihydroxybenzene; Pyrocathecol), C6H4(OH)2
110.11 [120-80-9]White crystals, which become discol-
ored on exposure to air and light. Readily soluble in water,
in alcohol, in benzene, in ether, in chloroform, and in pyri-
dine, forming clear solutions.
Use a suitable grade with a content of not less than 99%.
the column temperature is maintained at 280. The area of
Cation-Exchange ResinUse a suitable grade.
the C12H9N peak is not less than 95.5% of the total peak
area.
Dowex 50-W-X8-100, from Sigma-Aldrich, www.sigma-
Carbon Dioxide Detector TubeA fuse-sealed glass tube
aldrich.com.]
so designed that gas may be passed through it. Contains
Cation-Exchange Resin, Carboxylate (Sodium Form)
(50- to 100-mesh)Use a suitable grade.
tors hydrazine and crystal violet.
[NOTEA suitable grade is available as Bio-Rex 70 from
Measuring range: 0.01 to 0.3 Vol.%.
BioRad Laboratories, www.bio-rad.com.]
[NOTEAvailable from Draeger Safety, Inc.,
Cation-Exchange Resin, PolystyreneUse a suitable
www.draeger.com, or from Gastec Corp., www.gastec.co.jp,
grade.
distributed in the USA by www.nextteq.com.]
[NOTEA suitable grade is available as Dowex-50X2-100,
Carbon Disulfide, ChromatographicUse a suitable
grade.
Cation-Exchange Resin, Styrene-DivinylbenzeneA
Carbon Disulfide, CS [75-15-0]Use ACS reagent
strongly acidic, cross-linked sulfonated resin containing
grade.
about 2% of divinylbenzene. It is available in the hydrogen
Carbon Monoxide Detector TubeA fuse-sealed glass
form in the 50- to 100-, 100- to 200-, and 200- to
tube so designed that gas may be passed through it. Con-
400-mesh sizes. It can be regenerated to the hydrogen form
tains suitable absorbing filters and support media for the
by treating with a hydrochloric acid solution (5 in 100). For
indicators iodine pentoxide and selenium dioxide and fum-
satisfactory regeneration, a contact time of at least 30 min-
ing sulfuric acid.
utes is required after which it must be washed free of excess
acid. It is insoluble in water, in methanol, and in acetoni-
trile. Suitable for use in column chromatography.
Moisture content of fully regenerated and expanded resin [NOTEA suitable grade is available commercially, in
Transfer 10 to 12 mL of the resin (as received) to a flask, precoated plate form, with fluorescent indicator, from EMD
and convert it completely to the hydrogen form by stirring Chemicals, www.emdchemicals.com.]
with 150 mL of hydrochloric acid solution (5 in 100) for not Ceric Ammonium Nitrate, Ce(NO3)4 2NH4NO3
less than 30 minutes. Decant the acid, and wash the resin in 548.22Use ACS reagent grade.
the same manner with water until the wash water is neutral Ceric Ammonium Sulfate, Ce(SO4)2 2(NH4)2SO4
to litmus (pH 3.5). 2H2O632.55Yellow to yellowishorange crystals. Dis-
Transfer 5 to 7 mL of the regenerated resin to a glass solves slowly in water, but more rapidly when mineral acids
filtering crucible, and remove only the excess surface water are present. Use ACS reagent grade.
by very careful suction filtration. Transfer the conditioned Ceric Sulfate, Ce(SO4)2 with a variable amount of
resin to a tared weighing bottle, and weigh. Dry in a vac- water(anhydrous) 332.24 [13590-82-4]It may also
uum oven at a pressure of 50 mm of mercury at 100 to contain sulfates of other associated rare earth elements. Yel-
105 for 16 hours. Transfer from the vacuum oven to a des- low to orange-yellow crystals or crystalline powder. Practi-
iccator, cool to room temperature, and weigh again. The cally insoluble in cold water; slowly soluble in cold dilute
loss in weight is between 75% and 83%. mineral acids, but more readily soluble when heated with
these solvents.
Total wet volume capacityTransfer 3 to 5 mL of the
regenerated, undried (See Moisture content above) resin to a AssayWeigh accurately about 800 mg, transfer to a
5-mL graduated cylinder, and fill it with water. Remove any flask, add 25 mL of water and 3 mL of sulfuric acid, and
air bubbles from the resin bed with a stainless steel wire, warm until dissolved. Cool, and add 60 mL of a mixture of
and settle the resin to its minimum volume by tapping the 1 volume of phosphoric acid and 20 volumes of water. Add
graduated cylinder. Record the volume of the resin. 25 mL of potassium iodide solution (1 in 10), insert the
Transfer the resin to a 400-mL beaker. Add about 5 g of stopper in the flask, and allow to stand for 15 minutes. Re-
sodium chloride, and titrate, stirring well, with 0.1 N so- place the air over the solution with carbon dioxide, and
dium hydroxide to the blue endpoint of bromothymol blue while continuing the flow of carbon dioxide into the flask,
(pH 7.0). titrate the liberated iodine with 0.1 N sodium thiosulfate VS,
adding 3 mL of starch TS as the endpoint is approached.
(net mL NaOH N)/(mL of resin) = mEq/mL Each mL of 0.1 N sodium thiosulfate is equivalent to
33.22 mg of Ce(SO4)2. Not less than 80.0% is found.
The total wet volume capacity of the resin is more than 0.6
Chloride (Reagent test)Dissolve 1 g in a mixture of 5 mL
mEq per mL.
of nitric acid and 4 mL of water. Filter, if necessary, and
Wet screen analysisThe purpose of this test is to prop- dilute with water to 20 mL. To 10 mL of the dilution add
erly identify the mesh size of the resin. To obtain an accu- 1 mL of silver nitrate TS, allow to stand for 10 minutes, and
rate screen analysis would require a special apparatus and filter until clear. To the remaining 10 mL of test solution add
technique. 1 mL of silver nitrate TS: any turbidity produced does not
Add 150 mL of resin to 200 mL of water in an appropriate exceed that in a control prepared by adding 0.05 mg of Cl
bottle, and allow it to stand at least 4 hours to completely to the filtrate obtained from the first 10 mL of test solution
swell the resin. (0.01%).
Transfer, by means of a graduated cylinder, 100 mL of
Heavy metalsHeat 500 mg with a mixture of 10 mL of
settled and completely swollen resin to the top screen of a
water and 0.5 mL of sulfuric acid until solution is complete.
series of the designated U.S. Standard 20.3-cm brass
Cool, dilute with water to 50 mL, and bubble hydrogen sul-
screens. Thoroughly wash the resin on each screen with a
fide gas through the solution until it is saturated: the precip-
stream of water until the resin is completely classified, col-
itate that is formed is white or not darker than pale yellow.
lecting the wash water in a suitable container. Wash the
IronDissolve 100 mg in a mixture of 5 mL of water and
beads remaining on the respective screens back into the
2 mL of hydrochloric acid, warming if necessary, and cool.
100-mL graduate, and record the volume of settled resin on
Transfer to a glass-stoppered cylinder, dilute with water to
each screen. At least 70% of the resin will be within the
25 mL, and add 5 mL of ammonium thiocyanate TS and
specific mesh size.
25 mL of ether. Shake gently, but well, and allow the layers
[NOTEA suitable resin is Dowex 50WX2, produced by
to separate: any pink color in the ether layer is not darker
Dow Chemical Co. (www.dow.com) and also available
than that of a control, similarly prepared, containing
through Sigma-Aldrich (www.sigma-aldrich.com).]
0.02 mg of added Fe (0.02%).
Cation-Exchange Resin, Styrene-Divinylbenzene,
Cesium Chloride, CsCl168.36 [7647-17-8]A
Strongly AcidicUse a suitable grade.
white powder. Very soluble in water; freely soluble in meth-
anol; practically insoluble in acetone. Use a suitable grade.
Dowex 50-W-X8-100, from Sigma-Aldrich, www.sigma-
CetrimideSee Cetyltrimethylammonium Bromide.
aldrich.com.]
Cetyltrimethylammonium Bromide (CTAB; Cetrimide;
Cation-Exchange Resin, Sulfonic AcidUse a suitable
Hexadecyltrimethylammonium Bromide), C19H42BrN364.46
grade.
[57-09-0]Use a suitable grade. For any chromatographic
application, use a suitable grade with a content of not less
Amberlyst 15 or as Dowex 50-W-X2 from Sigma-
than 99.0%.
Aldrich, www.sigma-aldrich.com.]
Cetyltrimethylammonium Chloride, 25 Percent in
Cedar Oil (for clearing microscopic sections)
Water, C19H42ClN320.00Use a suitable grade.
[8000-27-9]A selected, distilled oil from the wood of the
Charcoal, Activated (Activated Carbon; Decolorizing Car-
red cedar, Juniperus virginiana Linn e (Fam. Pinaceae), should
bon) C12.01 [7440-44-0]A fine, black powder, which
be used for this purpose. Refractive index: about 1.504 at
is the residue from the destructive distillation of various or-
20. For use with homogeneous immersion lenses, a spe-
ganic materials, treated to increase its high capacity for ad-
cially prepared oil having a refractive index of 1.5150
sorbing organic coloring substances, as well as nitrogenous
0.0002 at 20 is required.
bases. Use Activated Charcoal (USP monograph).
Cellulose, ChromatographicUse a suitable grade.
Chenodeoxycholic Acid, C24H40O4392.57
[NOTEA suitable grade is available from EMD Chemicals,
[474-25-9]White to off-white powder.
www.emdchemicals.com.]
AssayWhen tested by thin-layer chromatography, with
Cellulose, MicrocrystallineUse Cellulose, Microcrystal-
the use of plates coated with chromatographic reversed-
line, FCC.
phase C18 mixture, a developing system consisting of 1 N
Cellulose Mixture, ChromatographicUse a suitable
acetic acid in methanol and 1 N acetic acid (19:1), and
grade.
sprayed with a mixture of sulfuric acid and methanol (1:1),
heated at 110 for 20 minutes, and examined visually and benzene, in chloroform, and in ether. Use ACS reagent
under long-wavelength UV light, a single spot is exhibited. grade.
4-Chlorobenzoic Acid, ClC6H4COOH156.57
[74-11-3]White, crystalline solid.
Chloramine T (Sodium p-Toluenesulfonchloramide),
C7H7ClNNaO2S 3H2O281.69 [7080-50-4]Use ACS AssayDissolve about 700 mg, accurately weighed, in a
reagent grade. mixture of 100 mL of hot alcohol and 50 mL of water. Ti-
Chlorine, Cl270.9 [7782-50-5]Greenish-yellow trate with 0.5 N sodium hydroxide VS, determining the
gas. High-purity grade available from most suppliers of spe- endpoint potentiometrically. Perform a blank determination,
cialty gases. and make any necessary correction. Each mL of 0.5 N so-
Chlorine Detector TubeA fuse-sealed glass tube so de- dium hydroxide is equivalent to 78.28 mg of ClC6H4COOH.
signed that gas may be passed through it and containing Not less than 98% is found.
SolubilityOne g dissolved in 25 mL of 0.5 N sodium
tor o-tolidine.
hydroxide yields a clear and complete solution.
Measuring range: 0.2 to 3 ppm. m-Chlorobenzoic Acid (3-Chlorobenzoic Acid),
[NOTEAvailable from Draeger Safety, Inc., C7H5ClO2156.57 [535-80-8]Use a suitable grade.
www.draeger.com, or from Gastec Corp., www.gastec.co.jp, 4-Chlorobenzophenone, C13H9ClO216.66
distributed in the USA by www.nextteq.com.] [134-85-0]Use a suitable grade.
m-Chloroacetanilide, C8H8ClNO169.61Off-white to 1-ChlorobutaneSee n-Butyl Chloride.
beige granules. 2-Chloroethanol (Ethylene Chlorohydrin), C2H5ClO80.51
[107-07-3]Use a suitable grade with a content of NLT
Assay
99%.
MOBILE PHASEPrepare a mixture of acetonitrile and water
2-Chloroethylamine Monohydrochloride, C2H6ClN
(22:3).
HCl115.99Off-white powder.
PROCEDUREInject about 20 L into a suitable liquid chro-
matograph (see Chromatography 621) equipped with a
254-nm detector and a 4.6-mm 15-cm column that con-
tains packing L1. The flow rate is about 1.5 mL per minute.
The area of the C8H8ClNO peak is not less than 99.9% of
of a phase consisting of 14% cyanopropylphenyl-86% di-
methylpolysiloxane; the injection port temperature is main-
p-Chloroacetanilide, C8H8ClNO169.61White or pale
tained at 150; the detector temperature is maintained at
yellow, needle-shaped crystals or crystalline powder. Insolu-
300; and the column temperature is maintained at 50 and
ble in water; soluble in alcohol and in ether.
programmed to rise 10 per minute to 200. The area of
SolubilityOne g dissolves in 30 mL of alcohol to form a the C2H6ClN HCl peak is not less than 99% of the total
clear solution. peak area.
Melting range 741: between 178 and 181. Melting range 741: between 150 and 246.
Chloroform, CHCl3119.38 [67-66-3]Use ACS rea- Residue on ignition (Reagent test): not more than
gent grade. 0.1%.
Chloroform, Alcohol-FreeUse a suitable grade that 1-Chloroadamantane, C10H15Cl170.68 [935-56-8]
does not contain alcohol as a stabilizer. White crystalline solid.
Chloroform, MethylSee Methyl Chloroform.
Chlorogenic Acid, C16H18O9354.31 [327-97-9]
White to off-white powder. Use a suitable grade.
AssayWhen tested by thin-layer chromatography (see The following conditions have been found suitable: a 0.25-
Chromatography 621) with the use of plates coated with mm 30-m capillary column coated with a 1-m layer of
chromatographic silica gel mixture and a developing system phase G2; the injection port temperature is maintained at
consisting of a mixture of butyl alcohol, water, and acetic 250; the detector temperature is maintained at 300; and
acid (60:25:15), and examined under short-wavelength UV the column temperature is maintained at 150 and pro-
light, a single spot is exhibited, with trace impurities. grammed to rise 10 per minute to 280. The area of the
Chloromethylated Polystyrene-Divinylbenzene Anion- C10H15Cl peak is not less than 97.5% of the total peak area.
exchange ResinSee Anion-exchange Resin, Chloromethyl- 2-Chloro-4-aminobenzoic AcidSee 4-Amino-
ated Polystyrene-Divinylbenzene. 2-chlorobenzoic Acid.
1-Chloronaphthalene (-Chloronaphthalene), C10H7Cl 5-Chloro-2-aminobenzophenoneSee 2-Amino-
162.62 [90-13-1]Colorless to light yellow liquid. 5-chlorobenzophenone.
3-Chloroaniline, C6H6ClN127.57 [108-42-9]Col- AssayUse a gas chromatograph equipped with a flame-
orless to light brown liquid. Soluble in acid and in most ionization detector. The following conditions have been
organic solvents; practically insoluble in water. found suitable: a 3.2-mm 1.83-m stainless steel column is
packed with 7% phase G2 on support S1A; the injection AssayInject an appropriate specimen into a gas chro-
port temperature is maintained at 250 and the detector matograph (see Chromatography 621) equipped with a
temperature at 310; and the column temperature is pro- flame-ionization detector, helium being used as the carrier
grammed to increase at a rate of 10 per minute from 50 gas. The following conditions have been found suitable: a
to 250. Not less than 90% of C10H7Cl is found, of which 0.25-mm 30-m capillary column coated with a 1-m layer
not more than 10% is 2-chloronaphthalene. of phase G2; the injection port temperature is maintained at
250; the detector temperature is maintained at 300; and Refractive index 831: between 1.6320 and 1.6340, at
the column temperature is maintained at 150 and pro- 20.
grammed to rise 10 per minute to 280. The area of the 4-Chloro-1-naphthol, C10H7ClO178.6 [604-44-4]A
C6H6ClN peak is not less than 99% of the total peak area. white to off-white powder, with a melting point between
118 and 120. Use a suitable grade. Store below 0. Refractive index 831: between 1.592 and 1.596 at
2-Chloronicotinic Acid, C6H4ClNO2157.55 20.
[2942-59-8]Off-white powder. p-Chloroaniline, (4-Chloroaniline), C6H6ClN127.57
[106-47-8]Use a suitable grade. AssayInject an appropriate volume into a gas chromat-
Chlorobenzene, C6H5Cl112.56 [108-90-7]Clear, ograph (see Chromatography 621) equipped with a flame-
colorless liquid. Insoluble in water; soluble in alcohol, in ionization detector, helium being used as the carrier gas.
The following conditions have been found suitable: a 0.25- sponding brand names, are published periodically in Phar-
mm 30-m capillary column coated with a 1-m layer of macopeial Forum as a guide for the chromatographer.]
phase G2; the injection port temperature is maintained at Chromatographic Silica GelSee Silica Gel, Chromato-
280; the detector temperature is maintained at 300; and graphic.
the column temperature is maintained at 180 and pro- Chromatographic Silica Gel MixtureSee Silica Gel Mix-
grammed to rise 10 per minute to 280. The area of the ture, Chromatographic.
C6H4ClNO2 peak is not less than 98% of the total peak area. Chromatographic Siliceous EarthSee Siliceous Earth,
2-Chloro-4-nitroaniline, 99%, C6H5ClN2O2172.57 Chromatographic.
[121-87-9]White to off-white powder. Chromatographic Siliceous Earth, SilanizedSee Sili-
ceous Earth, Chromatographic, Silanized.
Chromatographic Solvent HexaneSee Hexane, Solvent,
Chloroplatinic Acid, H2PtCl6 6H2O517.90
Chromatographic.
[18497-13-7]Use ACS reagent grade Chloroplatinic Acid
Chromium Potassium Sulfate Dodecahydrate,
Hexahydrate.
CrK(SO4)2 12H2O499.40 [10279-63-7]Use ACS rea-
5-Chlorosalicylic Acid, C7H5ClO3172.57
gent grade.
Chromium Trioxide, CrO399.99 [1333-82-0]Use
AssayWhen tested by thin-layer chromatography (see
ACS reagent grade.
Chromatography 621) with the use of plates coated with
Chromogenic Substrate for Amidolytic TestSynthetic
chromatographic silica gel mixture and a developing system
molecules consisting of tripeptides or tetrapeptides coupled
consisting of a mixture of cyclohexane, chloroform, and ace-
to a chromophore. The terminal amino acid is specific for
tic acid (14:4:2), and examined visually and under long-
the protease utilized. The synthetic peptides mimic the pep-
wavelength UV light or iodine spray, a single spot is exhib-
tide sequence (specific to the activated coagulation factor)
ited.
of the active site on the natural substrate. The coagulation
factor catalyzes the splitting of the chromophore (p-ni-
1-Chloro-2,2,2-trifluoroethylchlorodifluoromethyl
troaniline) from the peptide. The amount of release can be
EtherUse a suitable grade.
measured directly in a spectrophotometer, because the
Chlorotrimethylsilane (Trimethylsylyl Chloride),
maximum absorbance spectra of the bound and free chro-
C3H9ClSi108.64 [75-77-4]Clear, colorless to light yel-
mophores differ. The released chromophore is a colored
low liquid. Fumes when exposed to moist air.
compound; the complete substrate itself is colorless.
[CAUTIONIt reacts vigorously with water, alcohols, and
Molecular weights of the various substrates range from
other hydrogen donors. Store in tight glass containers.]
about 600 to 750 Da. Solubility in aqueous solutions can
Refractive index 831: between 1.3850 and 1.3890 at vary. Not all substrates are of equal sensitivity, and incuba-
20. tion periods may have to be extended.
Chlortetracycline HydrochlorideUse Chlortetracycline Chromotrope 2R, C16H10N2Na2O8S2468.4
Hydrochloride (USP monograph). [4197-07-3]Red powder or crystals. Use a suitable grade.
Cholestane, C27H48372.67 [481-21-0]Use a suita- Chromotropic Acid (4,5-Dihydroxy-2,7-
ble grade. naphthalenedisulfonic Acid), C10H8O8S2 2H2O356.33
Cholesterol, C27H46O386.66 [57-88-5]Use a suita- [148-25-4], for the anhydrous formUse a suitable grade.
ble grade (NFmonograph). Chromotropic Acid Disodium Salt (4,5-Dihydroxy-2,7-
Cholesteryl Benzoate, C34H50O2490.76 [604-32-0] naphthalenedisulfonic Acid, Disodium Salt), C10H6O8Na2S2
Use a suitable grade. 2H2O400.29 [5808-22-0]Use ACS reagent grade.
Cholesteryl n-HeptylateUse a suitable grade. Cinchonidine, C19H22N2O294.39 [485-71-2]White
Choline Chloride, HOCH2CH2N(CH3)3Cl139.62 crystals, crystalline or granular powder. Soluble in alcohol
[67-48-1]White crystals or crystalline powder. Very soluble and in chloroform; practically insoluble in water.
in water. Is hygroscopic. Store in tight containers.
AssayTransfer about 100 mg, previously dried at 105 50 mL of glacial acetic acid. Add a few drops of p-naphthol-
for 2 hours and accurately weighed, to a beaker, add 20 mL benzein TS, and titrate with 0.1 N perchloric acid VS. Per-
of water and 1 drop of aluminum chloride solution (1 in form a blank determination, and make any necessary correc-
10), and mix. Add slowly 20 mL of a freshly prepared, tion. Each mL of 0.1 N perchloric acid is equivalent to
filtered sodium tetraphenylborate solution (1 in 50), and al- 14.72 mg of C19H22N2O. Not less than 99.0% is found.
low the mixture to stand for 30 minutes with occasional
swirling. Pass through a medium-porosity, sintered-glass fil-
ter, and wash the beaker and the precipitate with four
10-mL portions of water. The weight of the precipitate, de-
Specific rotation 781: between 105 and 115, calcu-
termined after drying at 105 for 2 hours, and multiplied by
lated on the dried basis, determined in a solution in alcohol
0.3298, gives the equivalent weight of C5H14ClNO. Not less
containing 10 mg per mL.
than 99.5% is found.
Cinchonine, C19H22N2O294.39 [118-10-5]White
crystals, crystalline or granular powder. Slightly soluble in
Chromatographic Fullers EarthSee Fullers Earth, Chro-
chloroform, sparingly soluble in alcohol, and practically in-
matographic.
soluble in water.
Chromatographic n-HeptaneSee n-Heptane, Chromato-
graphic.
50 mL of glacial acetic acid. Add a few drops of p-naphthol-
Chromatographic Magnesium OxideSee Magnesium
benzein TS, and titrate with 0.1 N perchloric acid VS. Per-
Oxide, Chromatographic.
form a blank determination, and make any necessary correc-
Chromatographic ReagentsSee Reagents, Chromato-
tion. Each mL of 0.1 N perchloric acid is equivalent to
graphic Reagents.
14.72 mg of C19H22N2O. Not less than 99.0% is found.
[NOTEListings of the numerical designations for phases
(G), packings (L), and supports (S), together with corre- Loss on drying 731Dry it at 105 to constant weight:
Melting range 741: between 255 and 261. ble in alcohol and in acetone. Melts at about 220, with
decomposition.
Specific rotation 781: between +219 and +229, cal-
culated on the dried basis, determined in a solution in alco- Absorption maximumThe UV absorption spectrum of a 1
hol containing 50 mg per 10 mL. in 100,000 solution in alcohol shows a maximum at about
Citric AcidUse Citric Acid Monohydrate (USP mono- 238 nm.
graph).
Specific rotation 781: about +209, determined in a 1 in
Citric Acid, Anhydrous [77-92-9]Use Anhydrous Citric
100 solution in alcohol.
Acid (USP monograph).
Cotton, AbsorbentUse Purified Cotton (USP mono-
Cobalt Chloride (Cobaltous Chloride), CoCl2 6H2O
graph).
m-Cresol Purple, C21H18O5S382.43 [2303-01-7]
Cobalt Nitrate, Co(NO3)2 6H2O291.03
Cupric Acetate, Cu(C2H3O2)2 H2O199.65
Cobaltous Acetate (Cobalt Acetate), Co(C2H3O2)2
4H2O249.08 [71-48-7]Red, needlelike crystals. Solu-
Cupric Chloride, CuCl2 2H2O170.48
ble in water and in alcohol. Use ACS reagent grade.
[7447-39-4]Bluish-green deliquescent crystals. Freely solu-
Cobaltous ChlorideSee Cobalt Chloride.
ble in water; soluble in alcohol; slightly soluble in ether. Use
Coenzyme Q9 (Ubiquinone 45), C54H82O4795.2
ACS reagent grade.
[303-97-9]Yellow to yellow-orange powder. Clear yellow
Cupric Citrate ([Citrato(4-)]dicopper), Cu2C6H4O7315.18
solution, at a concentration of 1 mg per mL in a mixture of
chloroform and ethanol (9:1).
Cupric Nitrate [3251-23-8]Use ACS reagent grade Cu-
AbsorptivityIts absorptivity at 275 nm, in alcohol solu- pric Nitrate Hydrate.
tion, is about 1700. Cupric Nitrate Hydrate, Cu(NO3)2 2.5H2O232.59
CollagenUse a suitable grade. [3252-23-8]; Cu(NO3)2 3H2O241.60 [10031-43-3]Use
[NOTEA suitable grade is acid-soluble Collagen Type I ACS reagent grade.
from calf skin, and is commercially available from Sigma- [NOTEThis reagent is available containing either 2.5 or 3
Aldrich Corp., www.sigma-aldrich.com; catalog number molecules of water of hydration.]
C3511.] Cupric Sulfate, CuSO4 5H2O249.69Use ACS reagent
Rat Tail CollagenUse a suitable grade. grade.
[NOTEA suitable grade is available from BD Biosciences, Cupric Sulfate, Anhydrous, CuSO4159.61
www.bdbiosciences.com.] [7758-98-7]A white or grayish-white powder free from a
CollagenaseUse a suitable grade. blue tinge. Upon the addition of a small quantity of water,
[NOTEA suitable grade is commercially available as Col- it becomes blue. Soluble in water. Store in tight containers.
lagenase Type 2, CLS-2, from Worthington Biochemical
Corp., www.worthington-biochem.com.]
0.02 mg of Cl (0.002%).
Compactin, C23H34O5390.52 [73573-88-3]Use a
Substances not precipitated by hydrogen sulfideDeter-
suitable grade.
mine as directed for ACS reagent grade of Cupric Acetate:
the residue weighs not more than 6 mg (0.15%).
Cupriethylenediamine Hydroxide Solution, 1.0 MUse
Congo Red, C32H22N6Na2O6S2696.67 [573-58-0]A
a suitable grade.
dark red or reddish-brown powder. Decomposes on expo-
[NOTEA suitable grade is available from GFS Chemicals,
sure to acid fumes. Its solutions have a pH of about 8 to
www.gfschemicals.com.]
9.5. One g dissolves in about 30 mL of water. Is slightly
Cyanoacetic Acid, C3H3NO285.06 [372-09-8]
soluble in alcohol.
White to light yellow, crystalline solid. Very soluble in water.
not more than 3.0% of its weight.
25 mL of water and 25 mL of alcohol. Titrate with 0.1 N
Residue on ignitionAccurately weigh about 1 g, previ-
sodium hydroxide VS, determining the endpoint potentio-
ously dried at 105 for 4 hours, and place it in a porcelain
metrically. Perform a blank determination and make any
dish or crucible. Carefully ignite until well charred, cool, add
necessary corrections. Each mL of 0.1 N sodium hydroxide
2 mL of sulfuric acid, and carefully ignite until the residue is
is equivalent to 85.06 mg of C3H3NO2. Not less than 99% is
white or practically so. Cool, add 0.5 mL of sulfuric acid and
found.
1 mL of nitric acid, evaporate, and again ignite to constant
Cyanogen Bromide, BrCN105.92 [506-68-3]Col-
weight: the weight of the sodium sulfate so obtained is be-
orless crystals. Volatilizes at room temperature. Its vapors are
tween 20.0% and 24.0% of the weight of the dried speci-
highly irritating and very toxic. Melts at about 52. Freely
men taken.
soluble in water and in alcohol. Store in tight containers in
SensitivenessTo 50 mL of carbon dioxide-free water add a cold place.
0.1 mL of congo red solution (1 in 1000). The red color of
SolubilitySeparate 1-g portions dissolve completely in
the solution is changed to violet by the addition of 0.05 mL
10 mL of water and in 10 mL of alcohol, respectively, to
of 0.10 N hydrochloric acid and is restored by the subse-
yield colorless solutions.
quent addition of 0.05 mL of 0.10 N sodium hydroxide.
4-Cyanophenol (4-Hydroxybenzonitrile), C6H4CNOH
Coomassie Blue G-250 (Coomassie Brilliant Blue G-250,
119.12 [767-00-0]Use 95 percent reagent.
Serva Blue G), C47H48N3O7S2Na854.0 [6104-58-1]A
Cyclam (1,4,8,11-Tetraazacyclotetradecane), C10H24N4
dark blue powder. Soluble in water. Use a suitable grade.
200.33 [295-37-4]Use 98 percent reagent.
Store between 15 and 30.
-Cyclodextrin (Cyclomaltohexaose; Schardinger -Dex-
Coomassie Brilliant Blue R-250, C45H44N3O7S2Na
trin), C36H60O30972.86 [10016-20-3]Use a suitable
825.97 [6104-58-1]Brown powder.
grade with a content of not less than 98.0%.
Copper, CuAt. Wt. 63.546 [7440-50-8]Use ACS
[NOTEA suitable grade is available from Fluka, www.
reagent grade.
sigma-aldrich.com, catalog number 28705.]
Cortisone, C21H28O5360.44 [53-06-5]White, crys-
talline powder. Practically insoluble in water; sparingly solu-
[NOTEA suitable grade is available from either BD Biosci-
Add the following:
ences, www.bdbiosciences.com or Applied Biosystems,
www.appliedbiosystems.com.] L
-Cyclodextrin (-Schardinger dextrin;
Deuterated Methanol (Methanol-
12
C-d4, Methyl-
12
C-d3 al-
Cycloheptaamylose), C42H70O35 xH2O1134.98 (anhy-
cohol-d1)36.1 [811-98-3]The degree of deuteration is
drous) [68168-23-0]Use a suitable hydrate grade.L USP36
not less than 99.8%. Is a clear colorless liquid miscible with
Cyclohexane, C6H1284.16 [110-82-7]Use ACS rea-
water, with alcohol, and with methylene chloride; density at
gent grade.
20: 0.888 g/mL; refractive index at 20 (D-line): 1.326;
Cyclohexanol, C6H12O100.16 [108-93-0]A clear
boiling point 65.4 (760 mm Hg).
liquid. Freely soluble in water. Miscible with alcohol, with
Deuterated WaterSee Deuterium Oxide.
ethyl acetate, and with aromatic hydrocarbons.
Deuterium Chloride (Deutero Hydrochloric Acid), DCl
AssayWhen examined by gas-liquid chromatography,
37.47 [7698-05-7]Toxic gas. Use a suitable grade with a
using suitable gas chromatographic apparatus and condi-
degree of deuteration of NLT 99%.
tions, it shows a purity of not less than 98%.
Deuterium Oxide, D2O20.032 [7789-20-0]Use a
Melting temperature: about 23.
suitable grade having a minimum isotopic purity of 99.8
Specific gravity: about 0.962, at 20. atom % of deuterium.
(1,2-Cyclohexylenedinitrilo)tetraacetic Acid (trans-1, Deuterochloroform, CDCl3120.38Use a suitable
2-Diaminocyclohexane-N,N,N,N-tetraacetic Acid), grade.
C14H22N2O8 H2O364.35Use ACS reagent grade. Devardas Alloy (Devardas Metal) [8049-11-4]A gray
Cyclohexylmethanol, C7H14O114.19Use a suitable powder composed of 50 parts of copper, 45 parts of alumi-
grade. num, and 5 parts of zinc.
L-Cystine, HOOC(NH2)CHCH2SSCH2CH(NH2)COOH Dextran, High Molecular Weight [9004-54-0]A dex-
240.30 [58-89-3]A white, crystalline powder. Very tran molecular weight standard having a weight-average
slightly soluble in water; soluble in dilute mineral acids and molecular weight, MW, of 1 to 2 10
6
Da and a weight-
in solutions of alkali hydroxides; insoluble in alcohol and in average molecular weight to number-average molecular
other organic solvents. weight ratio, MW / MN, of 1.0 to 1.8.
[NOTEA suitable grade is available from American Poly-
Specific rotation 781: between 215 and 225, de-
mer Standards Corporation, www.ampolymer.com.]
termined in a 2 in 100 solution of test specimen, previously
Dextrin, (C6H10O5)n xH2O [9004-53-9]A white amor-
dried over silica gel for 4 hours, in dilute hydrochloric acid
phous powder. Slowly soluble in cold water; more readily
(1 in 10) at a temperature of 20.
soluble in hot water; insoluble in alcohol.
Loss on drying 731Dry it over silica gel for 4 hours: it
Insoluble matterBoil 1 g with 30 mL of water in a small
loses not more than 0.2% of its weight.
flask: the solution is colorless and clear, or not more than
opalescent.
DEAE-Agarose [57407-08-6]Agarose beads chemically
bonded with diethylaminoethane and suspended in a 20%
ethanol solution in water.
[NOTECommercially available as DEAE-Sepharose.] Residue on ignition (Reagent test)Ignite 1 g with 0.5 mL
Decanol (n-Decyl Alcohol), C10H22O158.28 of sulfuric acid: the residue weighs not more than 5 mg
[112-30-1]A clear, viscous liquid. Specific gravity: about (0.5%).
0.83 at 20. Solidifies at about 6.5. Insoluble in water; solu-
Chloride (Reagent test)Dissolve 3 g in 75 mL of boiling
ble in alcohol and in ether.
water, cool, dilute with water to 75 mL, and filter if neces-
AssayWhen examined by gas-liquid chromatography, sary. To 25 mL of the filtrate add 2 mL of nitric acid and
using suitable gas chromatographic apparatus and condi- 1 mL of silver nitrate TS, and allow to stand for 5 minutes:
tions, it shows a purity of not less than 99%. any turbidity produced is not greater than that of a control
Decyl Sodium Sulfate, C10H21NaO4S260.33White, containing 0.02 mg of added Cl (0.002%).
crystalline solid.
Sulfate (Reagent test, Method I)To a 25-mL portion of
AssayTransfer about 1 g, accurately weighed, to a suita- the filtrate from the preceding test add 0.5 mL of diluted
ble, tared crucible, moisten with a few drops of sulfuric acid, hydrochloric acid and 2 mL of barium chloride TS, and allow
and ignite gently to constant weight. Each mg of residue is to stand for 10 minutes: any turbidity produced is not
equivalent to 3.662 mg of C10H21NaO4S. Not less than 95% greater than that of a control containing 0.2 mg of added
is found. SO4 (0.02%).
Dehydrated AlcoholSee Alcohol, Dehydrated.
Alcohol-soluble substancesBoil 1 g with 20 mL of alcohol
Deoxyadenosine Triphosphate, C10H16N5O12P3491.18
for 5 minutes under a reflux condenser, and filter while hot.
Evaporate 10 mL of the filtrate on a steam bath, and dry at
[NOTEA suitable grade is available from either BD Bio-
105: the residue weighs not more than 5 mg (1%).
sciences, www.bdbiosciences.com or Applied Biosystems,
Reducing sugarsShake 2 g with 100 mL of water for
www.appliedbiosystems.com.]
10 minutes, and filter until clear. To 50 mL of the filtrate
Deoxycytidine Triphosphate, C9H16N3O13P3467.16
add 50 mL of alkaline cupric tartrate TS, and boil for 3 min-
utes. Filter through a tared filtering crucible, wash with
water, then with alcohol, and finally with ether, and dry at
ences, www.bdbiosciences.com or Applied Biosystems,
105 for 2 hours: the precipitate of cuprous oxide weighs
not more than 115 mg (corresponding to about 5% of re-
Deoxyguanosine Triphosphate, C10H16N5O13P3507.18
ducing sugars as dextrose).
Dextro Calcium PantothenateUse Calcium Pantothen-
ate (USP monograph).
ences, www.bdbiosciences.com, or Applied Biosystems,
Dextrose, Anhydrous, C6H12O6180.16Use ACS rea-
gent grade D-Glucose, Anhydrous.
Deoxyribonucleic Acid PolymeraseThermostable, re-
DiacetylSee 2,3-Butanedione.
combinant DNA polymerase. Use a suitable grade.
3,3-Diaminobenzidine Hydrochloride,
[NOTEA suitable grade is available from Applied Biosys-
(NH2)2C6H3C6H3(NH2)2 4HCl360.11 [7411-49-6]
tems, www.appliedbiosystems.com.]
White to yellowish-tan (occasionally purple), needle-shaped
Deoxythymidine Triphosphate, C10H17N2O14P3
crystals. Soluble in water. Stable in organic solvents but un-
482.17Use a suitable grade.
stable in aqueous solution at room temperature. Store aque- (made by dissolving 2.84 g of crystallized sodium borate in
ous solutions in a refrigerator. 90 mL of warm water, adding 8.2 mL of 1 N sodium hy-
droxide, and diluting with water to 100 mL) and 0.1 mL of
Insoluble matterDissolve 2 g in 100 mL of water, with-
a solution of 10 mg of the test specimen in 20 mL of alco-
out heating, and filter immediately: the insoluble residue
hol: a distinct blue color develops within 10 minutes.
does not exceed 1 mg (0.05%).
Dibutyl Phthalate, C16H22O4278.34 [84-74-2]
Residue on ignition (Reagent test): not more than 1 mg,
Clear, colorless liquid.
from 2 g (0.05%).
AssayAccurately weigh about 2 g into a suitable flask,
Suitability test for detection of seleniumDissolve 1.633 g
add 25.0 mL of 1 N sodium hydroxide and 30 mL of isopro-
of selenious acid (H2SeO3) in water, and dilute with water to
pyl alcohol, and mix. Digest the mixture at a temperature
1 L. Dilute 10 mL of this solution with water to 1 L, to make
near boiling for 30 minutes, then cool in a water bath to
a solution containing 0.010 mg of Se per mL. Place 1 mL of
room temperature. Add phenolphthalein TS, and titrate with
the resulting solution in a 100-mL beaker, add 2 mL of for-
1 N sulfuric acid VS to the disappearance of the pink color.
mic acid solution (1 in 7), and dilute with water to 50 mL.
Perform a complete blank determination, and make any
Add 2 mL of 3,3-diaminobenzidine hydrochloride solution
necessary correction. Each mL of 1 N sulfuric acid consumed
(1 in 200), and allow to stand for 30 to 50 minutes. Adjust
is equivalent to 139.2 mg of C16H22O4. Not less than 98% is
with 6 N ammonium hydroxide to a pH between 6 and 7.
found.
Transfer to a 125-mL separator, add 10.0 mL of toluene, and
shake vigorously for 30 seconds: a distinct yellow color is
produced in the toluene layer. A blank containing diami- Acid contentAccurately weigh about 10 g, and dissolve
nobenzidine hydrochloride but no selenium standard, in 100 mL of an alcohol-ether mixture (1:1). Add phenol-
treated in the same manner, shows no color in the toluene phthalein TS, and titrate immediately with 0.05 N alcoholic
layer. potassium hydroxide VS. Each mL of 0.05 N alcoholic potas-
2,3-Diaminonaphthalene, C10H10N2158.20 sium hydroxide is equivalent to 4.15 mg of phthalic acid:
[771-97-1]Use a suitable grade. not more than 0.02% is found.
Diatomaceous Earth [91053-39-3]Use a suitable grade. Dibutylamine, C8H19N129.24 [111-92-2]Colorless
[NOTEA suitable grade is available as Celite 545-AW.] liquid.
Diatomaceous Earth, Flux-Calcined [91053-39-3]Use a
suitable grade.
[NOTEA suitable grade is Chromosorb W, AW-DMCS,
available from Grace, www.grace.com.]
Diatomaceous Earth, Silanized [91053-39-3]Use a suit-
able grade.
[NOTESuitable grades are available commercially as
Anachrome Q, Gas-Chrom Q, and Varaport 30.]
Diatomaceous Silica, Calcined [68855-54-9]A form of
silica (SiO2) consisting of fused frustules and fragments of
C8H19N peak is not less than 99% of the total peak area.
diatoms. It is an amorphous, fine, light pink or white pow-
der. Insoluble in water, in acids, and in dilute solutions of
20.
alkali hydroxides.
Dibutylammonium PhosphateUse a suitable grade.
Loss on ignitionAccurately weigh about 4 g, and ignite
[NOTEA suitable grade is available as PIC Reagent D4
to constant weight: it loses not more than 10.0% of its
from Waters Corporation, www.waters.com.]
weight.
1,3-Dicaffeoylquinic Acid (Cynarin; (1R,3R,4S,5R)-1,3-Bis
Organic impuritiesIt does not darken appreciably upon [[3-(3,4-dihydroxyphenyl)propenoyl]oxy]-4,5-
ignition. dihydroxycyclohexanecarboxylic Acid), C25H24O12516.45
[NOTE A suitable grade is available as catalog number
3991 from www.chromadex.com.]
[NOTESuitable grades are Chromosorb P and
Dichloroacetic Acid, C2H2Cl2O2128.9 [79-43-6]
Chromosorb W, available from Grace, www.grace.com.]
Colorless liquid. Miscible with water, with alcohol, and with
Diaveridine (5-([3,4-Dimethoxyphenyl]methyl)-2,4-
ether. Use a suitable grade.
pyrimidinediamine), C13H16N4O2260.3 [5355-16-8]Use
2,5-Dichloroaniline, Cl2C6H3NH2162.02 [95-82-9]
a suitable grade. [NOTEA suitable grade is available as cat-
White, needle-like crystals. Slightly soluble in water; soluble
alog number 46174 from www.sigma-aldrich.com.]
in alcohol and in ether.
Dibasic Ammonium CitrateSee Ammonium Citrate, Di-
basic. Melting range, Class I 741: between 49 and 50.
Dibasic Ammonium PhosphateSee Ammonium Phos- 2,6-Dichloroaniline, C6H5Cl2N162.02 [608-31-1]
phate, Dibasic. Off-white powder.
Dibasic Potassium PhosphateSee Potassium Phosphate,
Dibasic.
o-Dichlorobenzene, C6H4Cl2147.00 [95-50-1]Clear
DibenzylSee Bibenzyl.
liquid, having a light yellowish-brown tint (about APHA 20).
2,6-Dibromoquinone-chlorimide (2,6-Dibromo-N-chloro-
Practically insoluble in water. Miscible with alcohol and with
p-benzoquinone Imine; DBQ Reagent), O:C6H2Br2:NCl
ether. Boils at about 180.
299.35 [537-45-1]A yellow, crystalline powder. Insolu-
AssayWhen examined by gasliquid chromatography,
ble in water; soluble in alcohol and in dilute alkali hydroxide
with the use of suitable apparatus and conditions, it shows a
solutions.
purity of not less than 98%.
Density: between 1.299 and 1.301.
Solubility in alcoholA solution of 100 mg in 10 mL of
alcohol is not more than faintly turbid.
25.
Residue on ignition (Reagent test)Ignite 500 mg with
Residue on evaporationEvaporate 80 mL on a steam
0.5 mL of sulfuric acid: the residue weighs not more than
bath, and dry at 105 for 1 hour: the residue weighs not
1 mg (0.2%).
SensitivenessTo 10 mL of a water solution containing
AcidityAdd phenolphthalein TS to 25 mL of methanol,
0.01 mg of phenol add 0.3 mL of a sodium borate buffer
and titrate with 0.02 N alcoholic potassium hydroxide VS
until a faint pink color persists for 15 seconds. Pipet 25 mL crystalline powder. Insoluble in water; soluble in alcohol and
of test specimen into the solution; mix, avoiding exposure in dilute alkali hydroxide solutions.
to the atmosphere; and titrate with 0.02 N alcoholic potas-
sium hydroxide VS: not more than 2.2 mL is required to
restore the pink color (about 0.005%).
alcohol is complete and clear.
1,2-DichloroethaneSee Ethylene Dichloride.
Residue on ignitionIgnite 500 mg with 0.5 mL of sulfuric
Dichlorofluorescein, C20H10Cl2O5401.20 [76-54-0]
acid: the residue weighs not more than 1 mg (0.2%).
[NOTEThis specification covers both the 4,5- and 2,7-iso-
SensitivenessIt meets the requirements of the test for mers of dichlorofluorescein, either of which is suitable for
Sensitiveness under 2,6-Dibromoquinone-chlorimide. the preparation of dichlorofluorescein TS.] A weak orange-
Dicyclohexyl, (Bicyclohexyl) C12H22166.31 colored, crystalline powder. Sparingly soluble in water; solu-
[92-51-3]Use a suitable grade. ble in alcohol and in solutions of alkali hydroxides.
Dicyclohexyl Phthalate, C20H26O4330.42 [84-61-7].
Melting range 741: between 62 and 66. 5 drops of sulfuric acid: the residue weighs not more than
Dicyclohexylamine, (C6H11)2NH181.32 [101-83-7] 1 mg (0.5%).
Clear, strongly alkaline liquid. Sparingly soluble in water.
SensitivenessDissolve 100 mg in 60 mL of alcohol, add
Miscible with common organic solvents. Use a suitable
2.5 mL of 0.1 N sodium hydroxide, and dilute with water to
grade with a content of not less than 98%.
100 mL. Add 1 mL of this solution to a solution of potas-
Diethylamine, (C2H5)2NH73.14 [109-89-7]Color-
sium iodide prepared by dissolving 100 mg of potassium io-
less, flammable, strongly alkaline liquid. Miscible with water
dide, previously dried at 105 to constant weight and accu-
and with alcohol. Forms a hydrate with water. May be irri-
rately weighed, in 50 mL of water containing 1 mL of glacial
tating to skin and mucous membranes. Store in well-closed
acetic acid, and titrate with 0.1 N silver nitrate VS until the
containers. Use ACS reagent grade.
color of the precipitate changes from pale yellowish-orange
Diethylamine Phosphate (Phosphoric Acid: Diethyl-
to pink. The volume of 0.1 N silver nitrate consumed is not
amine), C4H11N H3PO4171.13 [68109-72-8]Use a
more than 0.10 mL greater than the calculated volume, the
suitable grade. [NOTEA suitable grade is available from
calculated volume being based upon the KI content of the
www.richmanchemical.com.]
dried specimen as determined in the Assay under Potassium
N,N-Diethylaniline, C6H5N(C2H5)2149.23 [91-66-7]
Iodide (USP monograph).
Light yellow to amber liquid.
Dichlorofluoromethane, CHCl2F102.92 [75-43-4]
AssayInject an appropriate specimen (about 0.2 L) Colorless gas.
621) equipped with a flame-ionization detector, helium
being used as the carrier gas flowing at about 40 mL per
thermal-conductivity detector, helium being used as the car-
minute. The following conditions have been found suitable:
rier gas. The following conditions have been found suitable:
a 3-mm 1.8-m stainless steel column containing 20%
a 0.53-mm 30-m capillary column coated with a 5-m
phase G16 on support S1A; the injection port temperature
layer of phase G2; the injection port temperature is main-
is maintained at 250; the column temperature is main-
tained at 140 and programmed to rise 6 per minute to
200. The detector temperature is maintained at 310. The
programmed to rise 5 per minute to 40, and then to rise
area of the N,N-diethylaniline peak having a retention time
10 per minute to 180. The area of the CHCl2F peak is not
of about 4.9 minutes is not less than 99% of the total peak
less than 98% of the total peak area.
area.
2,6-Dichloroindophenol SodiumSee 2,6-Dichlorophe-
Refractive index 831: between 1.5405 and 1.5425 at nol-indophenol Sodium.
20. DichloromethaneUse Methylene Chloride.
Diethylene Glycol, (Bis(2-hydroxyethyl) Ether; Diglycol; 2,4-Dichloro-1-naphthol, C10H6OCl2213.06
2-Hydroxyethyl Ether; 2,2-Oxydiethanol), C4H10O3106.12 [2050-76-2]Light tan powder.
[111-46-6]A colorless to faintly yellow, viscous, hygro-
scopic liquid. Miscible with water, with alcohol, with ether,
and with acetone. Insoluble in benzene and in carbon tetra-
ceed 2.
chloride. Use a suitable grade with a content of not less
2,6-Dichlorophenol-indophenol Sodium (2,6-Dichloro-in-
than 99.0%.
dophenol Sodium), O:C6H2Cl2:NC6H4ONa with about 2H2O
Di(ethylene glycol) Methyl Ether (2-(2-Methoxyeth-
290.08 (anhydrous) [620-45-1] Use ACS reagent grade.
oxy)ethanol; Methyldiglycol), C5H12O3120.15
2,6-Dichlorophenylacetic Acid, C8H6Cl2O2205.04
[6575-24-2]White powder.
Diethylene Glycol Succinate Polyester,
(OCH2CH2OCH2CH2OOCCH2CH2COO)n [26183-02-8]
Clear, viscous liquid. Soluble in chloroform. Is stabilized by
modification of the diethylene glycol succinate polyester, to
render it suitable for use in gasliquid chromatography to a
temperature of 200.
C8H6Cl2O2 peak is not less than 97% of the total peak area.
2,6-Dichloroquinone-chlorimide (2,6-Dichloro-N-chloro-p-
benzoquinone Imine), O:C6H2Cl2:NCl210.44Pale yellow,
[NOTEA suitable grade is available from Alltech, www. any necessary correction. Each mL of 0.1 N sodium hydrox-
alltechweb.com.] ide is equivalent to 15.41 mg of C7H6O4. Not less than 99%
Diethylenetriamine, C4H13N3103.17 [111-40-0] is found.
Colorless liquid.
Melting range 741: about 207, with decomposition.
AssayInject an appropriate specimen into a suitable gas 2,7-DihydroxynaphthaleneSee 2,7-Naphthalenediol.
chromatograph (see Chromatography 621) equipped with Diiodofluorescein, C20H10I2O5584.10 [31395-16-1]
a flame-ionization detector, helium being used as the carrier Orange-red powder. Slightly soluble in water; soluble in al-
gas. The following conditions have been found suitable: a cohol and in solutions of alkali hydroxides.
0.25-mm 30-m capillary column coated with G2. The in-
Residue on ignitionIgnite 200 mg with 5 drops of sulfu-
jection port temperature is maintained at 200; the column
ric acid: the weight of the residue does not exceed 1.0 mg
(0.5%).
10 per minute to 250 and held there for 5 minutes; and
SensitivenessAccurately weigh about 100 mg of potas-
the detector temperature is maintained at 300. The area of
sium iodide, previously dried at 105 to constant weight,
the main peak is not less than 95% of the total peak area.
and dissolve it in 50 mL of water. Add 1 mL of
diiodofluorescein TS prepared from the test specimen and
20.
1 mL of glacial acetic acid, and titrate with 0.1 N silver ni-
Di(2-ethylhexyl)phthalate [Bis(2-ethylhexyl)phthalate],
trate VS until the color of the precipitate changes from
C24H38O4390.56 [117-81-7]Use a suitable grade.
brownish-red to a bluish-red. The volume of 0.1 N silver
Diethylpyrocarbonate, C6H10O5162.14
nitrate consumed is not in excess of 0.10 mL over the calcu-
[1609-47-8]Clear, colorless liquid. Use a suitable grade.
lated volume, based on the KI content of the dried potas-
Diethyl Sulfone (Ethyl Sulfone), (C2H5)2SO2122.19
sium iodide determined as follows. Dissolve about 500 mg
[597-35-3]Use a suitable grade with NLT 97%.
of potassium iodide, accurately weighed, in about 10 mL of
Digitonin, C56H92O291229.31 [11024-24-1]White,
water, and add 35 mL of hydrochloric acid and 5 mL of
crystalline powder. Almost insoluble in water; soluble in
chloroform. Titrate with 0.05 M potassium iodate VS until
warm alcohol, and in glacial acetic acid and in 75% acetic
the purple color of iodine disappears from the chloroform.
acid; insoluble in chloroform and in ether. Melts at about
Add the last portions of the iodate solution dropwise, agitat-
230, with decomposition.
ing vigorously and continuously. After the chloroform has
Specific rotation 781: between 47 and 49, deter- been decolorized, allow the mixture to stand for 5 minutes.
mined in a solution in 75% acetic acid containing 100 mg If the chloroform develops a purple color, titrate further
per mL. with the iodate solution. Each mL of 0.05 M potassium io-
date is equivalent to 16.60 mg of KI.
Diisodecyl Phthalate [Bis(isodecyl)phthalate], C28H46O4
warm alcohol is colorless and complete.
Diisopropyl Ether (Isopropyl Ether), [(CH3)2CH]2O
it loses not more than 6% of its weight.
102.17 [108-20-3]Colorless, mobile liquid. Slightly sol-
uble in water. Miscible with alcohol and with ether.
Digoxigenin (3,12,14,21-Tetrahydroxy-20(22)-
[CAUTIONIt is highly flammable. Do not use where it may
norcholenic Acid Lactone; 3,12,14-Trihydroxy-5,20(22)-
be ignited. Do not evaporate to the point of near dryness,
cardenolide; 5,20(22)-Cardenolide-3,12,14-triol;
since it tends to form explosive peroxides.]
Lanadigenin), C23H34O5390.51 [1672-46-4]Use a suita-
Specific gravity: between 0.716 and 0.720.
ble grade.
Distilling range, Method II 721Not less than 95% dis-
Dihydroquinidine Hydrochloride, C20H27ClN2O2362.89
tills between 65 and 70.
[1476-98-8]Rhombic plates. Freely soluble in methanol
and in chloroform. PeroxidesTo 10 mL, contained in a clean, glass-stop-
pered cylinder previously rinsed with a portion of the ether
Assay
under examination, add 1 mL of freshly prepared potassium
MOBILE PHASEPrepare a mixture of water, acetonitrile, di-
iodide solution (1 in 10). Shake, and allow to stand for
ethylamine, and methanesulfonic acid (860:100:20:20).
1 minute: no yellow color is observed in either layer (about
0.001% as H2O2).
Residue on evaporation[NOTEIf peroxide is present, do
not carry out this procedure.] Evaporate 14 mL (10 g) from a
tared shallow dish, and dry at 105 for 1 hour: the residue
The area of the C20H27ClN2O2 peak is not less than 97.5% of
AcidityAdd 2 drops of bromothymol blue TS to 10 mL
Dihydroquinine (Hydroquinine), C20H26N2O2326.43
of water in a glass-stoppered, 50-mL flask, and titrate with
[522-66-7]Freely soluble in acetone, in alcohol, and in
0.010 N sodium hydroxide until a blue color persists after
chloroform; almost insoluble in water.
vigorous shaking. Add 5 mL of diisopropyl ether, and titrate
Assay
with 0.010 N sodium hydroxide: not more than 0.30 mL is
MOBILE PHASEPrepare a mixture of water, acetonitrile, di-
required to restore the blue color (0.005% as CH3COOH).
ethylamine, and methanesulfonic acid (860:100:20:20) and
[NOTEFor spectrophotometric determinations, use di-
methanol (82:18).
isopropyl ether that meets the following additional
PROCEDUREInject about 20 L into a suitable liquid chro- requirement:]
AbsorbanceIts absorbance at 255 nm, in a 10-mm
quartz cell, does not exceed 0.2, water being used as the
blank.
The area of the C20H26N2O2 peak is not less than 97.5% of
Diisopropylamine, [(CH3)2CH]2NH101.19
2,5-Dihydroxybenzoic Acid, C7H6O4154.12
AssayNot less than 98% of C6H15N is found, a suitable
[303-07-1]Off-white powder. Freely soluble in alcohol
gas chromatograph equipped with a flame-ionization detec-
yielding a clear, very pale yellow solution.
tor being used. The following conditions have been found
suitable: a 3.2-mm 1.83-m stainless steel column is
30 mL of methanol. Slowly add 40 mL of water. Titrate with
packed with a cross-linked polystyrene support; the injection
0.1 N sodium hydroxide VS, determining the endpoint po-
port temperature is maintained at 250 and the detector
tentiometrically. Perform a blank determination, and make
temperature at 310; the column temperature is pro- The area of the dimethyl phthalate peak is not less than
grammed to rise at 10 per minute from 50 to 220. 99% of the total peak area.
Refractive index 831: between 1.3915 and 1.3935, at Refractive index 831: between 1.514 and 1.518 at 20.
20. Dimethyl Sulfone (Methyl Sulfone), (CH3)2SO294.13
Diisopropylethylamine (N,N-Diisopropylethylamine), [67-71-0]White crystals.
C8H19N129.24 [7087-68-5]Clear, colorless liquid. Sol-
uble in glacial acetic acid.
Dimethyl Sulfoxide (Methyl Sulfoxide), (CH3)2SO
AssayAccurately weigh about 500 mg, dissolve in 78.13Use ACS reagent grade methyl sulfoxide.
50 mL of glacial acetic acid, mix, add crystal violet TS, and Dimethyl Sulfoxide, Spectrophotometric Grade
titrate with 0.1 N perchloric acid VS. Each mL of 0.1 N [67-68-5]Use methyl sulfoxide ACS spectrophotometric re-
perchloric acid is equivalent to 12.92 mg of C8H19N. Not agent grade.
less than 98% is found. N,N-Dimethylacetamide, C4H9NO87.12
[127-19-5]Clear, colorless liquid. Miscible with water and
with many organic solvents. Use a suitable HPLC or spectro-
20.
scopic grade.
1,2-Dilinoleoyl-3-oleoyl-rac-glycerol, C57H100O6881.4
p-Dimethylaminoazobenzene (Methyl Yellow, Butter Yel-
low), C6H5N:NC6H4N(CH3)2225.29 [60-11-7]Yellow
1,2-Dilinoleoyl-3-palmitoyl-rac-glycerol, C55H98O6
leaflets or yellow, crystalline powder.
Diluted Acetic AcidSee Acetic Acid, Diluted. SolubilityInsoluble in water; sparingly soluble in chloro-
Diluted AlcoholUse Diluted Alcohol (NF monograph). form, in ether, or in fatty oils. Dissolve 100 mg in 20 mL of
Diluted Hydrochloric AcidSee Hydrochloric Acid, Di- alcohol: the solution is complete or practically so and clear.
luted.
Diluted Nitric AcidSee Nitric Acid, Diluted.
Diluted Sulfuric AcidSee Sulfuric Acid, Diluted.
SensitivenessAdd 0.05 mL of an alcohol solution (1 in
Dimethicone, viscosity 500 centistokes (Poly(dimethyl-
200) and 2 g of ammonium chloride to 25 mL of carbon
siloxane), viscosity 500 centistokes), [Si(CH3)2O]n
dioxide-free water: the lemon-yellow color of the solution is
changed to orange by the addition of 0.05 mL of 0.1 N
2,5-Dimethoxybenzaldehyde, C9H10O3166.17
hydrochloric acid and restored on the subsequent addition
[93-02-7]Off-white crystals.
of 0.05 mL of 0.1 N sodium hydroxide.
p-Dimethylaminobenzaldehyde,
(CH3)2NC6H4CHO149.19 [100-10-7]Use ACS reagent
a flame-ionization detector, nitrogen being used as the car-
grade.
p-Dimethylaminocinnamaldehyde,
a 0.3-mm 30-m capillary column coated with phase G1;
(CH3)2NC6H4CH:CHCHO175.23Orange-yellow powder.
the injection port temperature is maintained at 270; the
Soluble in acetone, in alcohol, and in benzene.
detector temperature is maintained at 300; the column
2-Dimethylaminoethyl Methacrylate, C8H15NO2
10 per minute to 270. The area of the main peak is not
less than 97% of the total peak area.
Dimethylaminophenol (meta isomer), C8H11NO
137.18Purplishblack, gray, or tan-colored, crystalline
1,2-Dimethoxyethane, C4H10O290.12 [110-71-4]
solid.
Clear, colorless liquid. Miscible with water and with alcohol.
Soluble in hydrocarbon solvents. May form peroxides on
2,6-Dimethylaniline, C8H11N121.18 [87-62-7]Yel-
standing.
low liquid.
Boiling range (Reagent test)Not less than 95% distills
between 83 and 86.
N,N-Dimethylaniline, C6H5N(CH3)2121.18
[121-69-7]Light yellow liquid. Clear, colorless liquid when
20.
freshly distilled, but acquiring a reddish to reddish-brown
AcidityTo 20 mL add bromophenol blue TS, and titrate
color. Specific gravity: about 0.960. Freezing point about 2.
with 0.020 N sodium hydroxide: not more than 2.0 mL is
Insoluble in water; soluble in alcohol, in chloroform, in
consumed (about 0.015% as CH3COOH).
ether, and in dilute mineral acids.
Water, Method I 921: not more than 0.2%.
AssayInject an appropriate specimen (about 0.2 L)
Dimethoxymethane (Formaldehyde Dimethyl Acetal, Meth-
ylal), C3H8O276.10 [109-87-5]Use a suitable grade.
621) equipped with a flame-ionization detector, helium
(3,4-Dimethoxyphenyl)acetonitrile (Homoveratronitrile),
being used as the carrier gas flowing at about 40 mL per
C10H11NO2177.20 [93-17-4]Off-white fibers.
minute. The following conditions have been found suitable:
Melting range 741: between 65 and 67. a 3-mm 1.8-m stainless steel column containing 20%
Dimethyl Phthalate, C10H10O4194.19 [131-11-3] phase G16 on support S1A; the injection port temperature
Viscous, colorless liquid. is maintained at 250; the column temperature is main-
Assay
200. The detector temperature is maintained at 310. The
MOBILE PHASEPrepare a filtered and degassed mixture of
area of the N,N-dimethylaniline peak having a retention
chromatographic n-heptane and n-propyl alcohol (HPLC
time of about 11.5 minutes is not less than 99% of the total
grade) (97:3). Make adjustments if necessary (see System
peak area.
Suitability under Chromatography 621).
STANDARD SOLUTIONDissolve a suitable quantity of di- Refractive index 831: between 1.5571 and 1.5591 at
methyl phthalate in Mobile phase to obtain a solution having 20.
a known concentration of about 0.26 mg per mL.
Boiling range (Reagent test)Distill 100 mL: the differ-
ence between the temperatures observed, when 1 mL and
95 mL have distilled, is not more than 2.5. Its boiling tem-
perature at a pressure of 760 mm of mercury is 194.2.
HydrocarbonsDissolve 5 mL in a mixture of 10 mL of Sulfanilamide testDissolve 20 mg of USP Sulfanilamide
hydrochloric acid and 15 mL of water: a clear solution re- RS in 100 mL of water to obtain the Sulfanilamide solution.
sults and it remains clear on cooling to about 10. Into two 150-mL beakers pipet 1.0 mL and 2.5 mL of the
Sulfanilamide solution, respectively. Dilute with water to
Aniline or monomethylanilinePlace 5 mL in a glass-stop-
90 mL. To provide a blank, place 90 mL of water in a third
pered flask, add 5 mL of a solution of acetic anhydride in
beaker. To each beaker add 8.0 mL of trichloroacetic acid
benzene (1 in 10), mix, and allow to stand for 30 minutes.
solution (3 in 20) and 1.0 mL of sodium nitrite solution (1
Add 30.0 mL of 0.5 N sodium hydroxide VS, shake the mix-
in 1000). Stir the solutions for 5 minutes, then add 10 mL of
ture, add phenolphthalein TS, and titrate with 0.5 N hydro-
acetate buffer TS, and 1.0 mL of a 1 in 1000 solution of N,
chloric acid VS. Perform a blank determination, and make
N-dimethyl-1-naphthylamine in alcohol. The pH is about 5
any necessary correction. Not more than 0.30 mL of 0.5 N
to 6, using pH paper. Stir for an additional 5 minutes, then
sodium hydroxide is consumed by the test specimen.
add 20 mL of glacial acetic acid. The pH is about 3 to 4,
3,4-Dimethylbenzophenone, C15H14O210.27
using pH paper. In comparison with the blank, the beaker
[2571-39-3]White chunks melting at about 45.
containing 1.0 mL of the Sulfanilamide solution shows a pink
color, while the other beaker shows a deep pink to red
color.
N,N-Dimethyloctylamine, C10H23N157.30
0.25-mm 30-m capillary column coated with phase G1:
the detector temperature and the injection port temperature
2,5-Dimethylphenol, C8H10O122.16 [95-87-4]Use
are maintained at 300; the column temperature is main-
a suitable grade.
tained at 180 and programmed to rise at the rate of 10
2,6-Dimethylphenol, (CH3)2C6H3OH122.16
per minute to 280 and held at that temperature for
[576-26-1]White to pale yellow, crystalline solid.
10 minutes. The area of the main peak is not less than 99%
of the total peak area. AssayInject a 1 in 3 solution of it in xylene into a suita-
5,5-Dimethyl-1,3-cyclohexanedione, C8H12O2140.18 ble gas chromatograph equipped with a flame-ionization
[126-81-8]White, crystalline solid. Slightly soluble in detector, helium being used as the carrier gas at a flow rate
water; soluble in alcohol, in methanol, in chloroform, and in of about 40 mL per minute. The following conditions have
acetic acid. been found suitable: a 3.2-mm 1.83-m stainless steel col-
umn packed with 10% phase G25 on support S1A; the in-
jection port temperature is maintained at about 250 and
N,N-Dimethyldecylamine (1-(Dimethylamino)decane),
the detector temperature at about 310; the column tem-
C4H9NO185.35 [1120-24-7]Use a suitable grade with
perature is programmed to rise at 8 per minute from 100
a content of not less than 98%.
to 200. Similarly inject a specimen of xylene. The area of
1,5-Dimethyl-1,5-diazaundecamethylene
the C8H10O peak is not less than 98% of the total peak area
polymethobromide (Polybrene), [28728-55-4]This is a
corrected for xylene.
positively charged polymer. It is available as an off-white,
crystalline solid or powder and is extremely hygroscopic. Melting range 741: between 44 and 46.
Use a suitable reagent grade. 3,5-Dimethylphenol, C8H10O122.16 [108-68-9]
[NOTECommercially available as Polybrene.] Use a suitable grade.
N,N-Dimethyldodecylamine-N-oxide (Lauryldimeth- [NOTEA suitable grade is available as product number
ylamine-N-oxide), CH3(CH2)11NO(CH3)2229.40 5600 from www.alfa.com.]
[1643-20-5]Use a suitable grade. [NOTEN,N- N,N-Dimethyl-p-phenylenediamine Dihydrochloride,
Dimethyldodecylamine-N-oxide, 0.1 N in Water and N,N- (CH3)2NC6H4NH2 2HCl209.12 [99-89-9]Nearly
Dimethyldodecylamine-N-oxide, 30 Percent are suitable white, fine, crystalline, hygroscopic solid that may have a
grades as well.] pinkish cast. Freely soluble in water; soluble in alcohol.
Dimethylethyl(3-hydroxyphenyl)ammonium Chloride
See Edrophonium Chloride.
250-mL beaker, and dissolve in about 75 mL of water. Ti-
Dimethylformamide (N,N-Dimethylformamide),
trate with 0.1 N sodium hydroxide VS, determining the
HCON(CH3)273.09 [68-12-2]Use ACS reagent grade.
endpoint potentiometrically. Each mL of 0.1 N sodium hy-
N,N-Dimethylformamide Diethyl Acetal147.22
droxide is equivalent to 10.46 mg of C8H12N2 2HCl. Not
less than 98% is found.
SolubilityA solution of 1 g in 10 mL of water produces
not more than a slight haze.
1,3-Dimethyl-2-imidazolidinone, C5H10N2O114.15
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl Tetrazolium
Bromide, C18H16N5SBr414.3 [298-93-1]Yellow to or-
1,9-Dimethyl-methylene Blue, C36H46Cl4N6OS2Zn
ange powder. Use a suitable grade.
850.1 [23481-50-7]Dark green powder. Use a suitable
Dimethyltin Dibromide, C2H6Br2Sn308.59
grade.
m-Dinitrobenzene, C6H4(NO2)2168.11 [99-65-0]
Pale yellow crystals or crystalline powder. Almost insoluble
N,N-Dimethyl-1-naphthylamine, C12H13N171.24
in cold water; slightly soluble in hot water. Soluble in chlo-
[86-56-6]Pale yellow to yellow, aromatic liquid. Soluble in
roform and in benzene; sparingly soluble in alcohol. Is vola-
alcohol and in ether.
tile in steam.
suitable beaker, add 100 mL of glacial acetic acid, and dis-
solve by stirring. When solution is complete, titrate with 0.1
3,5-Dinitrobenzoyl Chloride, C7H3ClN2O5230.56
N perchloric acid VS, determining the endpoint potentio-
[99-33-2]Pale yellow, crystalline powder. Freely soluble in
metrically. Perform a blank determination, and make any
dilute sodium hydroxide solutions; soluble in alcohol.
necessary correction. Each mL of 0.1 N perchloric acid is
[CAUTIONCorrosive, moisture-sensitive, lachrymator, and
equivalent to 17.12 mg of C12H13N. Not less than 98% is
possible mutagen. Store under nitrogen.]
found.
Solubility in sodium hydroxideA solution of 500 mg in 20, sodium light being used.
25 mL of 1 N sodium hydroxide is clear or not more than
faintly turbid.
Residue on ignitionIgnite 1 g with 0.5 mL of sulfuric Disodium Chromotropate (4,5-Dihydroxy-2,7-
acid: the residue weighs not more than 1 mg (0.1%). naphthalenedisulfonic Acid, Disodium Salt), C10H6O8S2Na2
2,4-Dinitrochlorobenzene, C6H3(NO2)2Cl202.55 2H2O400.29Use ACS reagent grade Chromotropic Acid
[97-00-7]Yellow to brownish-yellow crystals. Insoluble in Disodium Salt.
water; soluble in hot alcohol, in ether, and in benzene. Disodium EthylenediaminetetraacetateSee Edetate Di-
sodium.
Disodium PhosphateSee Sodium Phosphate.
Residue on ignitionIgnite 500 mg with 5 drops of sulfu-
5,5-Dithiobis(2-nitrobenzoic Acid) (3-Carboxy-4-ni-
ric acid: the residue weighs not more than 1 mg (0.2%).
trophenyl disulfide; Ellmans reagent), C14H8N2O8S2396.35
2,4-Dinitrofluorobenzene (1-Fluoro-2,4-dinitrobenzene),
[69-78-3]Yellow powder, melting at about 242. Sparingly
C6H3FN2O4186.10 [70-34-8]Light yellow solid. Use a
soluble in alcohol.
suitable grade.
Dithiothreitol (Clelands Reagent; Threo-1,4-dimercapto-
2,4-Dinitrophenylhydrazine, 2,4-C6H3(NO2)2NHNH2
2,3-butanediol; DTT), HSCH2CH(OH)CH(OH)CH2SH154.25
198.14 [119-26-6]Orange-red crystals, which under
[3483-12-3]Slightly hygroscopic needles from ether. Freely
the microscope appear individually to be lemon-yellow, lath-
soluble in water, in alcohol, in acetone, in ethyl acetate, and
like needles. Very slightly soluble in water; slightly soluble in
in ether.
alcohol; moderately soluble in dilute inorganic acids. Use a
suitable grade with a content of not less than 97%.
Dithizone (Diphenylthiocarbazone; Phenylazothioformic
Dioctyl Sodium SulfosuccinateUse Docusate Sodium.
Acid 2-Phenylhydrazide), C6H5N:NCSNHNHC6H5256.33
Dioxane (Diethylene Dioxide; 1,4-Dioxane), C4H8O288.11
1-Dodecanol (Dodecyl Alcohol), CH3(CH2)11OH186.33
Diphenyl Ether (Phenyl Ether), (C6H5)2O170.21
[112-53-8]A clear, colorless liquid. Crystallizes as leaflets
[101-84-8]A colorless liquid. Insoluble in water; soluble in
from dilute alcohol solution. Use ACS reagent grade.
glacial acetic acid and in most organic solvents. Boils at
Dodecyl AlcoholSee 1-Dodecanol.
about 259.
Dodecyl Lithium Sulfate (Lithium Dodecyl Sulfate, Lithium
Lauryl Sulfate), C12H25LiO4S272.3 [2044-56-6]White
Diphenylamine, (C6H5)2NH169.22 [122-39-4]Use
to off-white powder, clear to slightly hazy, colorless to faint
ACS reagent grade.
yellow solution in water at 50 mg per mL at ambient tem-
Diphenylborinic Acid, Ethanolamine Ester, (2-Ami-
perature. The UV absorbance of a 0.1 M solution is less than
noethyl Diphenylborinate) C14H16BNO225.09White, crys-
0.05 at both 260 and 280 nm. The pH of a 0.1 M solution
talline powder. Use a suitable grade.
in water is 7.0 0.5. Use a suitable grade.
Dodecyl Sodium Sulfonate (Sodium 1-Dodecanesulfonate;
Diphenylcarbazide, (C6H5NHNH)2CO242.28
1-Dodecanesulfonic Acid, Sodium Salt), C12H25SO3Na272.38
[140-22-7]Use ACS reagent grade 1,5-Diphenyl-
[2386-53-0]White, powdery solid. One g dissolves in
carbohydrazide.
50 mL of warm water to yield a clear, colorless solution. Use
Diphenylcarbazone [Diphenylcarbazone compd. with
a suitable grade.
s-Diphenylcarbazide (1:1)], C6H5NHNHCON:NC6H5
Melting point 741: higher than 300.
C6H5NHNHCONHNHC6H5482.54 [538-62-5]Use ACS
3-(Dodecyldimethylammonio)propanesulfonate (Lauryl
reagent grade Diphenylcarbazone Compound with
sulfobetaine, N,N-dimethyl-N-dodecyl-N-(3-sulfopropyl)
s-Diphenylcarbazide (1:1).
ammonium betaine), C17H37NO3S335.54
2,2-Diphenylglycine, C14H13NO2227.26
[3060-50-2]Off-white powder. Melts at about 244, with
Dodecyltriethylammonium Phosphate, 0.5 M,
decomposition.
[C12H25N (C2H5)3]3PO4906.52Use a suitable grade.
[NOTEA suitable grade is available as Ion Pair Cocktail
30 mL of methanol. Slowly add about 20 mL of water, heat-
Q12 (catalogue number 404031) from Regis Technologies,
ing slightly if necessary for complete solution. Titrate with
Inc., www.registech.com.]
0.1 N sodium hydroxide VS, determining the endpoint po-
Dodecyltrimethylammonium Bromide
tentiometrically. Perform a blank determination and make
(Lauryltrimethylammonium bromide), CH3(CH2)11N(CH3)3Br
any necessary correction. Each mL of 0.1 N sodium hydrox-
ide is equivalent to 22.73 mg of C14H13NO2. Not less than
98.0% is found.
D5047 from Sigma-Aldrich, www.sigma-aldrich.com.]
DipicrylamineSee Hexanitrodiphenylamine.
Drabkins ReagentThe reagent consists of 100 parts of
Dipropyl Phthalate, C14H18O4250.29 [131-16-8]
sodium bicarbonate, 20 parts of potassium ferricyanide, and
Viscous, colorless liquid.
5 parts of potassium cyanide. [CAUTIONThe reagent is
Assay HIGHLY TOXIC. Very toxic by inhalation, in contact with
skin, and if swallowed, and there is a risk of serious damage MOBILE PHASEPrepare a mixture of acetonitrile and water
to eyes. Wear suitable protective clothing, gloves, and eye (52:48).
and face protection. Do not mix with acids. Contact with
acids liberates a very toxic gas. If ingested, perform gastric
lavage, and call a physician.]
[NOTEThe reagent can be obtained from many manu-
facturers and suppliers. Some examples of manufacturers or
The area of the C14H18O4 peak is not less than 99% of the
suppliers are the following: Sigma Chemical Co., St. Louis,
total peak area.
MO; and CIMA Scientific, Dallas, TX.]
Dried PeptoneSee Peptone, Dried.
4,4-Dipyridyl, C10H8 N2156.18 [553-26-4]Use a
Earth, Chromatographic, Silanized, Acid-Base
suitable grade. [NOTEA suitable grade is available from
WashedUse a suitable grade.
Sigma-Aldrich Catalog number 289426.]
[NOTEA suitable chromatographic grade is Gas-Chrom
,-DipyridylSee 2,2-Bipyridine.
Q, available from Alltech, www.alltechweb.com.]
Edetate Disodium (Disodium Ethylenediaminetetraacetate)
C10H14N2O8Na2 2H2O372.24Use ACS reagent grade
(Ethylenedinitrilo)tetraacetic Acid Disodium Salt Dihydrate.
n-Eicosane, C20H42282.55 [112-95-8]White, crys- [NOTESuitable peroxide test strips can be obtained from
talline solid. EMD Chemicals, www.emdchemicals.com, or from J. T.
Baker, www.mallbaker.com.]
Ethidium Bromide, C21H20N3Br394.3 [1239-45-8]
Eicosanol [629-96-9]Use a suitable grade.
Purple to purple-red powder. Use a suitable grade.
Eosin Y (Eosin Yellowish Y) (Certified Biological Eosin Y;
4-Ethoxyacetophenone, C10H12O2164.20
Sodium Tetrabromofluorescein), C20H6Br4Na2O5691.85
[1676-63-7]White to tan crystals.
[17372-87-1]Red to brownish-red pieces or powder. Use
ACS reagent grade. AssayDissolve about 50 mg in 1 mL of ether. Inject
Epiandrosterone (trans-Androsterone, 5-Androstan-3-ol- about 1 L of this solution into a suitable gas chromato-
17-one), C19H30O2290.44 [481-29-8]Use a suitable graph (see Chromatography 621) equipped with a flame-
grade. ionization detector and a 0.25-mm 30-m capillary column
[NOTEA suitable grade is available as catalog number containing stationary phase G1. The carrier gas is helium.
E3375 from www.sigma-aldrich.com.] The chromatograph is programmed as follows. Initially, the
Equilenin, C18H18O2266.33 [517-09-9]Colorless or column temperature is equilibrated at 180, then the tem-
white crystals or crystalline powder. Insoluble in water; solu- perature is increased at a rate of 10 per minute to 280,
ble in chloroform and in dioxane; moderately soluble in al- and maintained at 280 for 10 minutes. The injection port
cohol. temperature is maintained at 280, and the detector is
maintained at 300. The area of the 4-ethoxyacetophenone
Melting range, Class II 741: between 256 and 260.
peak is not less than 97.5% of the total peak area.
mined in a solution in dioxane containing 75 mg of equi-
2-Ethoxyethanol (Ethylene Glycol Monoethyl Ether),
lenin in each 10 mL.
C4H10O290.12 [110-80-5]Clear, colorless liquid. Mis-
Absorption maximaAn alcohol solution exhibits absorp-
cible with water, with alcohol, with ether, and with acetone.
tion maxima at 231, 282, 325, and 340 nm.
-Ergocryptine, C32H41N5O5575.70 [511-09-1]Use
a suitable grade. Boiling range (Reagent test)Not less than 95% distills
Eriochrome Black TSodium Chloride IndicatorMix between 133 and 135.
0.1 g of eriochrome black T and 10 g of sodium chloride, Ethyl Acetate, CH3COOC2H588.11 [141-78-6]Use
and triturate until the mixture becomes homogenous. ACS reagent grade.
Eriochrome Cyanine R, C23H15Na3O9S536.40 Ethyl Acrylate [140-88-5]Use a suitable grade.
[3564-18-9]Dark, red-brown powder. Freely soluble in Ethyl Alcohol (Alcohol; Ethanol), C2H5OH46.07Use Al-
water; insoluble in alcohol. cohol.
Ethyl Arachidate, C22H44O2340.6 [18281-05-5]Use
Solubility200 mg in 100 mL of water yields a solution
a suitable grade.
that remains clear and free from undissolved matter for
Ethyl Benzoate, C9H10O2150.17 [93-89-0]Clear,
30 minutes.
colorless liquid. Practically insoluble in water; miscible with
Loss on drying 731Dry it in vacuum over silica gel to
alcohol, with chloroform, and with ether.
constant weight: it loses not more than 2% of its weight.
Residue on ignition (Reagent test)0.5 g, treated with
chromatograph (see Chromatography 621), helium being
1 mL of sulfuric acid and 2 mL of nitric acid, yields between
used as the carrier gas. The following conditions have been
42.0% and 44.0% of the dry weight (theoretical yield is
found suitable: a 3-mm 2.4-m stainless steel column con-
42.9% of Na2SO4).
taining 20% phase G16 on support S1A; the injection port,
SensitivenessAdd 2 mL of a solution (1 in 1000) to 1 mL
column, and detector temperatures are maintained at 180,
of aluminum sulfate solution (1 in 10,000), heat at 37 3
195, and 250, respectively. The area of the ethyl benzoate
for 5 minutes, cool, and add 1 mL of sodium acetate TS: a
peak is not less than 98% of the total peak area.
strong red to red-violet color is produced in not more than
5 minutes.
20.
Escin [6805-41-0]Use a suitable grade.
Ethyl Cyanoacetate, CNCH2COOC2H5113.11
Ethanesulfonic Acid, C2H5SO3H110.13 [594-45-6]
[105-56-6]Colorless to pale yellow liquid. Slightly soluble
Colorless to light yellow liquid. Soluble in water.
in water. Miscible with alcohol and with ether. At atmos-
AssayAccurately weigh about 300 mg, dissolve in
pheric pressure it boils between 205 and 209, with de-
30 mL of water, add phenolphthalein TS, and titrate with
composition. At a pressure of 10 mm of mercury it distills at
about 90.
droxide is equivalent to 11.013 mg of C2H5SO3H: between
Specific gravity 841: between 1.057 and 1.062.
94.0% and 106.0% is found.
AcidityDissolve 2 mL in 25 mL of neutralized alcohol,
add phenolphthalein TS, and titrate with 0.10 N sodium
20.
hydroxide: not more than 1.5 mL is required to produce a
EtherSee Ethyl Ether.
pink color.
Ether, AbsoluteSee Ethyl Ether, Anhydrous.
Ethyl Ether (Diethyl Ether; Ether), (C2H5)2O74.12
Ether, DiphenylSee Diphenyl Ether.
Ether, IsopropylSee Diisopropyl Ether.
Ethyl Ether, Anhydrous (Diethyl Ether, Anhydrous; Ether,
Ether, Nonyl Phenyl Polyethylene GlycolSee (p-tert-
Absolute), (C2H5)2O74.12 [60-29-7]Use ACS reagent
Octylphenoxy)nonaethoxyethanol.
grade.
Ether, Peroxide-Free (Diethyl Ether; Ether), (C2H5)2O
Ethyl Salicylate, C9H10O3166.17 [118-61-6]Color-
74.12Use ACS reagent grade.
less liquid.
PeroxideTransfer 8 mL of potassium iodide and starch
TS to a 12-mL ground glass-stoppered cylinder about
15 mm in diameter. Fill completely with the substance
under test, mix, and allow to stand protected from light for
5 minutes. No color develops. Alternatively, peroxide test
strips may be used.
of methylsilicone; the injection port temperature is main-
300; the column temperature is maintained at 150 and
programmed to rise 10 per minute to 250. The area of
the ethyl salicylate peak is not less than 99% of the total potentiometrically, using a silver-ion selective electrode and
peak area. a calomel reference electrode containing 1 M potassium ni-
trate. Perform a blank determination, and make any neces-
sary correction. Each mL of 0.1 N silver nitrate is equivalent
20.
to 29.92 mg of C12H14IN: not less than 97.0% is found.
2-Ethylaminopropiophenone Hydrochloride,
Factor Xa (Activated Factor X) for Anti-Factor Xa Test
C6H5COCH(CH3)NHC2H5 HCl213.70 [51553-17-4]
Factor Xa is the proteolytic enzyme derived from bovine
plasma, and it cleaves prothrombin to form thrombin. It is a
4-Ethylbenzaldehyde, C2H5C6H4CHO134.18
glycoprotein having a molecular weight of 40,000. One Fac-
[4748-78-1]Colorless to pale yellow liquid.
tor Xa Unit is the amount of activated endogenous Factor X
AssayDissolve about 600 mg, accurately weighed, in a
(55,000 Da when nonactivated) that is contained in 1 mL of
mixture of 100 mL of alcohol and 25 mL of 1 M hydroxyla-
normal plasma. Russells viper venom is used to activate the
mine hydrochloride in a beaker. Cover the beaker with a
enzyme, and is then removed. The preparation is stabilized
watch glass. Heat gently until condensate begins to form on
and lyophilized.
the watch glass. Allow to cool for about 30 minutes. Titrate
Factor Xa is free of thrombin at the concentration used in
with 0.5 N sodium hydroxide VS, determining the endpoint
the test. When tested against pure fibrinogen, no clotting
potentiometrically. Perform a blank determination, and
takes place within 24 hours. It has not less than 40 Factor Xa
make any necessary correction. Each mL of 0.5 N sodium
Units per mg of protein, and exhibits 90% homogeneity
hydroxide is equivalent to 67.09 mg of C2H5C6H4CHO. Not
when tested by disk electrophoresis.
less than 98% is found.
One Factor Xa Unit per mL gives a 15-second clotting
Ethylbenzene, C8H10106.17 [100-41-4]Not less
time when tested in the following manner. Mix 0.1 mL of a
than 99.5%.
saturated solution of cephalin (derived from rabbit brain-
Ethylene Dichloride (1,2-Dichloroethane), C2H4Cl298.96
acetone powder or from an equivalent amount of rabbit
[107-06-2]Use ACS reagent grade 1,2-Dichloroethane.
brain thromboplastin) in 0.1 mL of citrated bovine plasma
Ethylene Glycol, HOCH2CH2OH62.07 [107-21-1]
and 0.1 mL of 0.025 M calcium chloride. Immediately add
Clear, colorless, slightly viscous, hygroscopic liquid. Slightly
0.1 mL of a solution of factor Xa (1 in 10), and incubate at
soluble in ether; practically insoluble in benzene. Miscible
37.
with water and with alcohol.
Fast Blue B Salt, C14H12N4O2 ZnCl4475.47
[91-91-8]Green powder.
Loss on drying 731Dry it in vacuum at 110 for
Residue on ignitionEvaporate 100 mL (110 g) in a tared 1 hour: it loses not more than 5.0% of its weight.
evaporating dish over a flame until the vapors continue to
AbsorbanceDissolve 50 mg in 100 mL of water. In a
burn after the flame is removed. Allow the vapors to burn
second container dissolve 100 mg of 2-naphthol in 100 mL
until the specimen is consumed. Ignite at 800 25 for
of 2-methoxyethanol. Pipet 5 mL of the test solution and
1 hour, cool, and weigh: the residue weighs not more than
10 mL of the 2-naphthol solution into a 100-mL volumetric
5.5 mg (0.005%).
flask, and dilute with acetone to volume. For the blank, pi-
AcidityAdd 0.2 mL of phenol red TS to 50 mL of water, pet 5 mL of water and 10 mL of 2-naphthol solution into a
and titrate with 0.1 N sodium hydroxide to a red endpoint. second 100-mL volumetric flask, and dilute with acetone to
Add 50 mL (55 g) of ethylene glycol, and titrate with 0.1 N volume. Determine the absorbance of the test solution in a
sodium hydroxide: not more than 1 mL is required to re- 1-cm cell at the wavelength of maximum absorbance at
store the red color (0.01% as CH3COOH). about 545 nm, with a suitable spectrophotometer, using the
blank to set the instrument: the absorbance is not less than Chloride (Reagent test)A 4.5-mL (5-g) portion shows
0.80. not more than 0.025 mg of Cl (5 ppm).
Fast Blue BB Salt, (C17H18ClN3O3)2 ZnCl2831.89
Water, Method I 921: not more than 0.20%.
[15710-69-7]Yellow powder melting at about 162, with
Ethylene Glycol Monoethyl EtherSee 2-Ethoxyethanol.
decomposition. Sparingly soluble in water.
Ethylene Oxide in Methylene Chloride (50 mg/mL)
ChlorideTransfer about 80 mg, accurately weighed, to a Use a suitable grade.
suitable beaker. Add 25 mL of acetone, 25 mL of water, and [NOTEA suitable grade is available from Sigma Aldrich
500 mg of sodium nitrate. Stir until solution is complete. Corporation, www.sigma-aldrich.com.]
Titrate with 0.01 N silver nitrate VS, determining the end Ethylenediamine (1,2-Diaminoethane) C2H8N260.10
point potentiometrically. Perform a blank determination and [107-15-3]Use a suitable grade with a content of not less
make any necessary correction. Not less than 15.0% of chlo- than 99%.
ride is found. N-Ethylmaleimide (1-Ethyl-1H-pyrrole 2,5-Dione),
Fast Green FCF, C37H34N2Na2O10S3808.86 C6H7NO2125.12 [128-53-0]Use a suitable grade.
[2353-45-9]Red to brown-violet powder or crystals. Solu-
ble in water; sparingly soluble in alcohol. Use a suitable
2-Ethyl-2-methylsuccinic Acid, C7H12O4160.17White
grade.
crystals.
FD&C Blue No. 1 (Brilliant Blue), C37H34N2O9S3Na2
suitable beaker, add 30 mL of methanol, and dissolve by
Ferric Ammonium CitrateThin, transparent, garnet-red
stirring. Slowly add 40 mL of deionized water. [NOTEIf
scales or granules or brownish-yellow powder. Is deliques-
cloudiness appears when adding the deionized water, dis-
cent and is affected by light. Very soluble in water; insoluble
solve in 50 mL of methanol only.] When solution is com-
in alcohol.
plete, titrate with 0.1 N sodium hydroxide VS, determining
AssayAccurately weigh about 1 g, dissolve in 25 mL of
the endpoint potentiometrically. Perform a blank titration,
water in a glass-stoppered flask, add 5 mL of hydrochloric
and make any necessary correction. Each mL of 0.1 N so-
acid and 4 g of potassium iodide, insert the stopper in the
dium hydroxide is equivalent to 16.02 mg of C7H12O4. Not
flask, and allow to stand in the dark for 15 minutes. Add
less than 98.5% of C7H12O4 is found.
100 mL of water, and titrate the liberated iodine with 0.1 N
Melting point 741: between 101 and 103.
sodium thiosulfate VS, adding 3 mL of starch TS as the
1-Ethylquinaldinium Iodide, C12H14IN299.15
endpoint is approached. Perform a blank determination, and
[606-55-3]Yellow-green solid. Sparingly soluble in water.
make any necessary correction. Each mL of 0.1 N sodium
thiosulfate is equivalent to 5.585 mg of Fe: between 16.5%
100 mL of water, and add 10 mL of glacial acetic acid. Ti-
and 18.5% is found.
trate with 0.1 N silver nitrate VS, determining the endpoint
Ferric citrateTo 250 mg dissolved in 25 mL of water add ZincTo the combined acid extracts saved from the Cop-
1 mL of potassium ferrocyanide TS: no blue precipitate is pertest, add 0.5 M sodium acetate to bring the pH between
formed. 5.0 and 5.5, and then add 1 mL of 0.1 N sodium thiosul-
fate. Add 10 mL of Standard Dithizone Solution (see Lead
TartrateDissolve 1 g in 10 mL of water, add 1 mL of
251), shake for 2 minutes, and allow the layers to separate.
potassium hydroxide TS, boil to coagulate the ferric hydrox-
Draw off the dithizone, and discard the water layer. Any
ide, adding more potassium hydroxide TS, if necessary, to
pink color is not greater than that in a control prepared by
precipitate all of the iron, filter, and slightly acidify the fil-
adding 0.006 mg of zinc ion (Zn) to the combined acid
trate with glacial acetic acid. Add 2 mL of glacial acetic acid,
extracts from the control used in the test for Copper
and allow to stand for 24 hours: no crystalline white precipi-
(0.005%).
tate is formed.
NitrateDissolve 10 g in 100 mL of dilute sulfuric acid (1
Lead 251Dissolve 1.0 g in 30 mL of water, add 5 mL
in 100), heat to boiling, and pour, slowly, into a mixture of
of dilute nitric acid (1 in 21), boil gently for 5 minutes, cool,
140 mL of water and 50 mL of stronger ammonia TS. Filter
and dilute with water to 50 mL: 20 mL of the solution
through a folded filter while still hot, wash with hot water
shows not more than 0.008 mg of Pb (0.002%).
until the volume of the filtrate is 300 mL, mix, and cool. To
Ferric Ammonium Sulfate, FeNH4(SO4)2 12H2O
15 mL of this solution add 1 mL of sodium chloride solution
(1 in 200), 0.10 mL of indigo carmine TS, and 15 mL of
Ferric Chloride, FeCl3 6H2O270.29 [10025-77-1]
sulfuric acid. The blue color is not completely discharged at
the end of 5 minutes (0.01%).
Ferric Nitrate, Fe(NO3)3 9H2O404.00
[10421-48-4]Use ACS reagent grade. Substances not precipitated by ammoniaEvaporate to
Ferric Sulfate, Fe2(SO4)3 xH2O [10028-22-5]Grayish- dryness 30 mL of the filtrate obtained in the test for Nitrate,
white, hygroscopic powder, or fawn-colored pearls, slowly and ignite gently: the weight of residue does not exceed
soluble in water. 1 mg (0.10%).
Ferrocyphen (Dicyano-bis (1,10-phenantroline) iron (II)
AssayAccurately weigh about 700 mg, and dissolve it in
Complex), (C12H8N2)2Fe(CN)2468.3Brown to black pow-
a mixture of 50 mL of water and 3 mL of hydrochloric acid
der. Dark red solution at 10 mg per mL in glacial acetic
in a glass-stoppered flask. Add 3 g of potassium iodide, and
acid. Soluble in chloroform and in water at 5 mg per mL,
allow to stand in the dark for 30 minutes. Then dilute with
yielding a clear purple solution in chloroform and a clear
100 mL of water, and titrate with 0.1 N sodium thiosulfate
orange solution in water.
VS, adding 3 mL of starch TS as the endpoint is approached.
Each mL of 0.1 N sodium thiosulfate is equiv- Water, Method I 921: not more than 10%.
alent to 5.585 mg of Fe: not less than 21.0% and not more Ferrous Ammonium Sulfate, Fe(NH4)2(SO4)2 6H2O
than 23.0% is found. 392.14Use ACS reagent grade.
Ferrous Sulfate, FeSO4 7H2O278.02 [7782-63-0]
Insoluble matter (Reagent test)A 10-g portion, dissolved
in a mixture of 100 mL of water and 5 mL of sulfuric acid,
Ferulic Acid (4-Hydroxy-3-methoxycinnamic acid),
shows not more than 2 mg of insoluble matter (0.02%).
ChlorideDissolve 1 g by warming with a mixture of
Fibroblast Growth Factor-2Use a suitable grade.
10 mL of water and 1 mL of nitric acid, add 4 mL of addi-
[NOTEA suitable grade can be obtained from Roche Di-
tional nitric acid, and dilute with water to 50 mL. To 25 mL
agnostics Corporation, www.roche-diagnostics.com.]
add 1 mL of phosphoric acid and 1 mL of silver nitrate TS.
Filter Paper, QuantitativeFor the Mercuric Bromide Test
Any turbidity does not exceed that produced in a control
Paper used in testing for arsenic, use Swedish O filter paper
containing 0.01 mg of chloride ion (Cl), 1 mL of nitric acid,
or other makes of like surface, quality, and ash.
1 mL of phosphoric acid, and 1 mL of silver nitrate TS
Fluorene, C13H10166.22 [86-73-7]White to off-
(0.002%).
white crystals or powder. Soluble in benzene, in carbon di-
Ferrous ironDissolve 4 g by warming with 50 mL of di-
sulfide, in ether, and in hot alcohol; freely soluble in glacial
lute sulfuric acid (1 in 10), cool, and titrate with 0.1 N
acetic acid.
potassium permanganate: not more than 0.16 mL is re-
Solubility testOne g dissolves in 10 mL of acetone to
quired to produce a permanent pink color (0.02% as Fe+2).
yield a clear and complete solution.
[NOTEBecause the reagents used in the tests for Copper
Melting range 741: between 113 and 117, within a 2
and Zinc may contain excessive amounts of copper and
range.
zinc, they should first be purified by extracting with
9-Fluorenylmethyl Chloroformate, C15H11ClO2258.70
Dithizone Extraction Solution(see Lead 251).]
[28920-43-6]Clear, colorless solid. Melts at about 62.
CopperDissolve 1.2 g in 100 mL of water. To 10 mL add
Fluorescamine, C17H10O4278.26 [38183-12-9]
50 mL of a solution containing 5 g of ammonium tartrate
White to off-white powder. Very slightly soluble in water;
and 5 mL of ammonium hydroxide. Add 10 mL of Standard
freely soluble in methylene chloride; soluble in alcohol;
Dithizone Solution (see Lead 251), shake for 2 minutes,
slightly soluble in chloroform.
draw off the dithizone layer, and compare the pink color
AssayDissolve about 600 mg in 75 mL of dimethylform-
with that in a control containing 6 g of copper ion (Cu)
amide, and titrate with 0.1 N lithium methoxide to a blue
and treated exactly as the 10-mL portion of test solution. If
endpoint, using 1% thymol blue in dimethylformamide as
the color in the test solution is less than that in the control,
the indicator. Perform a blank determination, and make any
then the test specimen contains less than the limit of both
necessary correction. Each mL of 0.1 N lithium methoxide is
Copper and Zinc. If the color in the test solution is more
equivalent to 27.83 mg of C17H10O4. Not less than 99% is
than that in the control, add 15 mL of dilute hydrochloric
found.
acid (1 in 250), and shake for 2 minutes. Draw off the
dithizone solution, and shake with a second 15 mL of dilute Loss on drying 731Dry it at 105 for 4 hours: it loses
hydrochloric acid (1 in 250) for 2 minutes. Draw off the not more than 0.5% of its weight.
dithizone, combine the two acid extracts, and reserve for 4-Fluoroacetophenone, FC6H4COCH3138.14
the Zinc test. Any pink color in the dithizone solution is not [403-42-9]Colorless liquid.
darker than that in the control solution treated exactly as
the test solution (0.005%).
AssayInject an appropriate specimen into a suitable gas Specific rotation 781: between +3.8 and +4.8, deter-
chromatograph (see Chromatography 621) equipped with mined in a solution of pyridine containing 10 mg per mL,
a flame-ionization detector, helium being used as the carrier with the use of a mercury light at 546.1 nm; between +21
gas. The following conditions have been found suitable: a and +25, determined in a solution of equal parts of chloro-
25-mm 30-m capillary column coated with a 1-m layer form and methanol containing 5 mg per mL, with the use
of phase G2; the injection port temperature is maintained at of sodium light.
SuitabilityDissolve 10 mg each of USP Digitoxin RS, pre-
viously dried, USP Digoxin RS, previously dried, and gitoxin,
respectively, in separate 5-mL portions of a mixture of
FC6H4COCH3 peak is not less than 99% of the total peak
2 parts of chloroform and 1 part of methanol, and dilute
area.
each with additional solvent mixture to 10 mL. Then pro-
Refractive index 831: 1.510 at 20. ceed as directed in the Identification test under Digoxin. The
Formaldehyde Solution, HCHO(30.03) and water chromatogram of gitoxin shows one fluorescent spot, lo-
Use ACS reagent grade. cated between the digoxin and digitoxin spots.
Formamide, HCONH245.04 [75-12-7]Use ACS re- Glacial Acetic AcidSee Acetic Acid, Glacial.
agent grade. Glass WoolFine threads of glass.
Preparation for Digitoxin AssayTo ensure freedom from Acid-soluble substancesBoil 1 g for 30 minutes with
ammonia, treat Formamide as follows. Shake a suitable 30 mL of diluted hydrochloric acid, and filter. Evaporate the
quantity of formamide with about 10% of its weight of an- filtrate, and dry the residue at 105 to constant weight: the
hydrous potassium carbonate for 15 minutes, and filter. Dis- residue weighs not more than 5 mg (0.5%).
till the filtrate in an all-glass apparatus under vacuum at a
Heavy metalsBoil 2 g with a mixture of 25 mL each of
pressure of about 25 mm of mercury or less. Reject the first
diluted nitric acid and water for 5 minutes, and filter. Evapo-
portion of distillate containing water, and collect the frac-
rate one-half of the filtrate to dryness, dissolve the residue in
tion that boils at about 115 at a pressure of 25 mm of
10 mL of water to which 3 drops of hydrochloric acid have
mercury or at 101 at a pressure of 12 mm of mercury.
been added, filter if necessary, and add an equal volume of
Store in tight containers, protected from light.
hydrogen sulfide TS to the filtrate: no darkening is pro-
Formamide, Anhydrous, HCONH245.04
duced.
[75-12-7]Use formamide that has a water content of less
D-Gluconic Acid, 50 Percent in Water, C6H12O7196.16
than 0.1%.
[526-95-4]Pale yellow liquid.
Formic Acid, HCOOH46.03 [64-18-6]Use ACS rea-
AssayDilute about 200 mg of the solution, accurately
gent grade Formic Acid, 88 Percent.
weighed, with 30 mL of water. Titrate with 0.1 N sodium
Formic Acid, 96 Percent, HCOOH46.03 [64-18-6]
hydroxide VS, determining the endpoint potentiometrically.
Use ACS reagent grade Formic Acid, 96 Percent.
Perform a blank determination and make any necessary cor-
Formic Acid, AnhydrousUse ACS reagent grade Formic
rection. Each mL of 0.1 N sodium hydroxide is equivalent to
Acid, 96 Percent.
19.62 mg of C6H12O7. Not less than 49.0% is found.
Fuchsin, Basic (Basic Red 9, Parafuchsin Hydrochloride),
C19H17N3323.82 [569-61-9]Use a suitable grade.
20.
Fullers Earth, Chromatographic(Very Fine and Moder-
Specific rotation 781: between +9.9 and +11.9, deter- ately Coarse)Gray or grayish-white powder or granules
mined as is, at 20. consisting mainly of hydrous aluminum-magnesium silicate.
Glucose, C6H12O6180.2 [50-99-7]Use a suitable
Powder finenessSee Powder Fineness 811.
grade. A white, crystalline powder. Freely soluble in water;
Soluble matterTwenty g, treated with 50 mL of cold
sparingly soluble in alcohol.
water and filtered, yields not more than 60 mg of residue
D-Glucuronolactone, C6H8O6176.12 [32449-92-6]
upon evaporation of the filtrate (0.3%). A second 20-g por-
tion, treated with 50 mL of cold alcohol and filtered, yields
Glutamic Acid, C5H9NO4147.13 [56-86-0]Use a
not more than 14 mg upon evaporation of the filtrate
suitable grade.
(0.07%).
L-Glutamic Acid, C5H9NO4147.1 [56-86-0]White
powder or white powder with a faint yellow cast. Use a
between 7.0% and 10.0% of its weight.
suitable grade.
[NOTEAdjust the water content, if necessary, by drying
L-Glutamine, C5H10N2O3146.15 [56-85-9]White,
in vacuum at room temperature, restoring the water re-
crystalline powder. Use a suitable grade.
quired, and equilibrating by shaking for 2 hours.]
Glycerin (Glycerol) [56-81-5]Use ACS reagent grade
Fuming Nitric AcidSee Nitric Acid, Fuming.
Glycerol.
Fuming Sulfuric AcidSee Sulfuric Acid, Fuming.
Glycolic Acid, C2H4O376.05 [79-14-1]White crys-
Furfural (2-Furancarboxyaldehyde; 2-Furaldehyde),
talline powder or chunks.
C4H3OCHO96.08 [98-01-1]Use ACS reagent grade.
Assay
G DesignationsSee phases for gas chromatography
SILYLATING REAGENT: Pyridine, hexamethyldisilazane, chloro-
under Reagents, Chromatographic Columns.
trimethylsilane (9:3:1)
Gadolinium (Gd III) Acetate Hydrate, (CH3CO2)3Gd
SAMPLE PREPARATION: Weigh approximately 25 mg (about xH2O334.38 [100587-93-7]White, crystalline, hygro-
3 drops) of the sample into a test tube. Add 2 mL of the scopic powder. Irritant. Use a suitable grade.
Silylating reagent and cap the test tube. Mix well and allow Geneticin (G418; O-2-Amino-2,7-dideoxy-D-glycero-alpha-
to incubate at room temperature for a minimum of 2 h. A D-glucoheptopyranosyl-(1-4)-O-(3-deoxy-4-C-methyl-3-(methyl-
white precipitate of ammonium chloride will form. Centri- amino)-beta-L-arabinopyranosyl-(1-6)-D-streptamine),
fuge and use the clear supernatant for injection. C20H40N4O10496.55 [49863-47-0]Use a suitable grade,
cell culture tested. PROCEDURE: Inject an appropriate volume of the Sample
Girard Reagent TSee Trimethylacethydrazide Ammonium preparation into a gas chromatograph (see Chromatography
Chloride. 621) equipped with a flame-ionization detector, helium
Gitoxin, C41H64O14780.94 [4562-36-1]White, crys- being used as the carrier gas. The following conditions have
talline powder. Practically insoluble in water, in chloroform, been found suitable: a 0.25-mm 30-m capillary column
and in ether; slightly soluble in pyridine and in diluted alco- coated with a 1-m layer of phase G2. The injection port
hol. Melts at about 250, with decomposition. temperature is maintained at 250; the detector tempera-
ture is maintained at 300; the column temperature is main-
tained at 100 and programmed to rise 10 per min to [NOTEA suitable grade is available as Bovine Hemoglobin
250. The area of the peak corresponding to C2H4O3 is NLT substrate powder from Sigma-Aldrich, www.sigma-aldrich.
98.5% of the total peak area. com.]
Gold Chloride (Chlorauric Acid), HAuCl4 3H2O393.83 1-Heptadecanol, C17H36O256.48 [1454-85-9]Use
[16903-35-8]Use ACS reagent grade. a suitable grade.
Guaiacol (o-Methoxyphenol), C7H8O2124.14 Heptafluorobutyric Acid, C4F7O2H214.04
[95-05-1]Colorless to yellowish, refractive liquid. Soluble [375-22-4]Use a suitable grade.
in about 65 parts of water; soluble in sodium hydroxide so- Heptakis-(2,6-di-O-methyl)--cyclodextrin (2,6-Di-O-
lution; miscible with alcohol, with chloroform, with ether, methyl--cyclodextrin; Dimethyl--cyclodextrin) C56H98O35
and with glacial acetic acid. 1331.36 [51166-71-3]Use a suitable grade.
n-HeptaneUse n-Heptane, Chromatographic.
AssayWhen examined by gasliquid chromatography, it
n-Heptane, Chromatographic C7H16100.21
[142-82-5]Clear, colorless, volatile, flammable liquid con-
tions have been found suitable for assaying it: a 3-mm
sisting essentially of C7H16. Practically insoluble in water; sol-
1.8-m stainless steel column containing liquid phase G16 on
uble in absolute alcohol. Miscible with ether, with chloro-
60- to 80-mesh support S1A. Helium is the carrier gas, the
form, with benzene, and with most fixed and volatile oils.
injection port temperature is maintained at 180, the col-
Use a suitable grade, chromatographic or HPLC, with a con-
umn temperature is maintained at 200, and the flame-ioni-
tent of not less than 99%.
zation detector is maintained at 280. The retention time is
Heptyl p-Hydroxybenzoate (Heptyl 4-Hydroxybenzoate;
about 8 minutes.
N-Heptyl 4-Hydroxybenzoate; Benzoic Acid, 4-Hydroxy-, Heptyl
Ester), C14H20O3236.31 [1085-12-7]Use a suitable
20.
grade with a content of NLT 98%.
Guanidine Hydrochloride, (Aminoformamidine Hydrochlo-
Hexadecyl Hexadecanoate (Hexadecyl Palmitate; Cetyl
ride; Aminomethanamidine Hydrochloride), CH5N3 HCl
Palmitate), C32H64O2480.85 [540-10-3]Use a suitable
95.53 [50-01-1]White, crystalline powder. Freely solu-
grade.
ble in water and in alcohol. Use a suitable grade with a
[NOTESuitable grades are available commercially as
content not less than 99%.
Hexadecyl Palmitate and Palmitic Acid Palmityl Ester from
Guanidine Isothiocyanate, C2H6N4S118.2
Sigma-Aldrich, www.sigma-aldrich.com, and Cetyl Palmitate,
[593-84-0]White powder or colorless crystals. Use a suita-
Catalog number C1203, from Spectrum Chemical Mfg.
ble grade.
Corp., www.spectrumchemical.com.]
Guanine Hydrochloride, C5H5N5O HCl H2O205.60
Hexadecyltrimethylammonium BromideSee Cetyl-
[635-39-2]White, crystalline powder. Melts above 250,
trimethylammonium Bromide.
with decomposition. Slightly soluble in water and in alcohol;
Hexadimethrine Bromide, (C13H30Br2N2)n [28728-55-4]
soluble in acidulated water and in sodium hydroxide TS. Its
White to off-white powder, hygroscopic, amorphous poly-
solutions are not precipitated by iodine TS or by
mer. Soluble in water up to 10% to give a colorless to light
mercuricpotassium iodide TS, but form a precipitate with
yellow solution. Use a suitable grade.
trinitrophenol TS.
Hexamethyldisilazane, C6H19NSi2161.39
[999-97-3]Clear, colorless liquid.
100 mg.
AssayWhen examined by gasliquid chromatography, it
tions have been found suitable for assaying the article: A
Hematein, C16H12O6300.26 [475-25-2]Prepared
2-mm 1.8-m glass column packed with phase G3 on sup-
from logwood extract or from hematoxylin by treatment
port S1. Helium, flowing at a rate of about 40 mL per min-
with ammonia and exposure to air. Reddish-brown crystals
ute, is the carrier gas; the detector temperature is about
with a yellowish-green metallic luster. Very slightly soluble in
310; the injection port temperature is about 100; and the
water (about 1 in 1700); slightly soluble in alcohol and in
column temperature is programmed to start at 35, hold for
ether; insoluble in benzene and in chloroform; freely soluble
5 minutes, then rise at a rate of 8 per minute to 200. A
in diluted ammonia solution to form a solution of dusky
flame-ionization detector is employed.
purplish-red color and in an aqueous solution of sodium hy-
Residue after evaporationTransfer 200 g to a tared dish,
droxide (1 in 50), to form a solution of bright red color,
and evaporate on a steam bath to dryness. Dry the residue
viewed in each case through a layer 1 cm in depth. Melts at
at 105 for 1 hour, cool, and weigh: not more than
a temperature above 200 and tends to decompose at 250.
0.0025% of residue is found.
Hematoxylin (Hydroxybrasilin), C16H14O6 3H2O356.32
Hexamethyleneimine (Homopiperidine), C6H12NH99.17
[517-28-2]A crystalline substance derived from the heart-
[111-49-9]Colorless to nearly colorless liquid.
wood of Haematoxylon campechianum Linn e (Fam. Legumi-
nosae). Colorless to yellow prisms. Very slightly soluble in
20.
cold water and in ether; rapidly soluble in hot water and in
HexamethylenetetramineSee Methenamine.
hot alcohol. When exposed to light, it acquires a red color
n-Hexane, C6H1486.18 [110-54-3](for use in spectro-
and yields a yellow solution. Dissolves in ammonia TS and in
photometry)Use Hexanes.
solutions of alkali hydroxides and carbonates. When dis-
Hexane, Solvent (Petroleum Benzin; Petroleum Ether, Lig-
solved in solutions of the following salts, it develops the
roin) [8032-32-4]Clear, volatile liquid. Practically insoluble
colors indicated: in alum solution, a red color; in stannous
in water; soluble in absolute alcohol. Miscible with ether,
chloride solution, a rose color; and in solutions of cupric
with chloroform, with benzene, and with most fixed and
salts, a greenish-gray color. It gradually turns black in potas-
volatile oils.
sium dichromate solution. Store hematoxylin and its solu-
[CAUTIONIt is dangerously flammable. Keep it away from
tions protected from light and air.
flames, and store in tight containers in a cool place.]
Hemoglobin, Bovine [9008-02-0]Use a suitable grade.
Use ACS reagent grade Petroleum Ether.
Hexane, Solvent, ChromatographicUse ACS HPLC rea-
gent grade.
Hexanes (suitable for use in UV spectrophotometry); usu-
ally a mixture of several isomers of hexane (C6H14), predomi-
nantly n-hexane, and methylcyclopentane (C6H12)Use ACS Use this grade also for alkoxyl determinations in assays in
spectrophotometric reagent grade. the individual monographs.]
Hexanitrodiphenylamine (Dipicrylamine), C12H5N7O12 Hydrobromic Acid, HBr80.91 [10035-10-6]Use
439.21 [131-73-7]Yellow-gold powder or prisms. Explo- ACS reagent grade.
sive. Usually contains about 15% of water as a safety pre- Hydrochloric Acid, HCl36.46 [7647-01-0]Use ACS
caution. Insoluble in water, in alcohol, in acetone, and in reagent grade.
ether; soluble in glacial acetic acid and in alkalies. Hydrochloric Acid, Diluted (10 percent) [7647-01-0]
Prepare by mixing 226 mL of hydrochloric acid with suffi-
Water, Method I 921: not more than 16%.
cient water to make 1000 mL.
Hexanophenone, C12H16O176.25 [942-92-7]Yel-
Hydrofluoric Acid, HF20.01 [7664-39-3]Use ACS
low liquid.
reagent grade.
Hydrogen Peroxide, H2O234.01 [7722-84-1]Use
ACS reagent grade with an assay content between 29.0%
and 32.0%.
Hydrogen Peroxide, 10 Percent, H2O234.01Dilute
30 mL of 30 percent hydrogen peroxide with water to
100 mL.
Hydrogen Peroxide, 30 Percent, Unstabilized H2O2
34.01 [7722-84-1]Use ACS reagent grade, with an assay
content between 29.0% and 32.0%, without an added sta-
C12H16O peak is not less than 98% of the total peak area.
bilizer.
Refractive index 831: 1.511 0.002 at 20.
Hydrogen Peroxide, 50 Percent in Water, H2O234.01
Hexylamine (1-Aminohexane), C6H15N101.19
Hydrogen Peroxide SolutionUse Hydrogen Peroxide
than 99%.
Topical Solution.
Histamine Dihydrochloride, C5H9N3 2HCl184.07Use
Hydrogen Sulfide, H2S34.08 [7783-06-4]Color-
USP Histamine Dihydrochloride RS.
less, poisonous gas, heavier than air. Soluble in water. Is
L-Histidine Hydrochloride Monohydrate, C6H9N3O2
generated by treating ferrous sulfide with diluted sulfuric or
HCl H2O209.63 [5934-29-2]Use a suitable grade.
diluted hydrochloric acid. Other sulfides yielding hydrogen
Horseradish Peroxidase Conjugated to Goat Anti-
sulfide with diluted acids may be used. Is also available in
Mouse IgGAffinity purified polyclonal antibody to Mouse
compressed form in cylinders.
Immune globulin (IgG) heavy and light chains (whole IgG)
Hydrogen Sulfide Detector TubeA fuse-sealed glass
produced in Goat and labeled with horseradish peroxidase.
tube so designed that gas may be passed through it and
Available either as a lyophilized powder or as a solution in a
containing suitable absorbing filters and support media for
suitable buffer, generally 10 mM sodium phosphate, pH 7.4,
the indicator, the latter consisting of a suitable lead salt.
containing a suitable preservative, such as 0.01% thimero-
sal, and an inactive protein(s) to prevent adsorption on the
[NOTEAvailable from Draeger Safety, Inc., www.draeger.
surface of the container. Use a suitable grade. Store at 20.
com, or from Gastec Corp., www.gastec.co.jp, distributed in
Hydrazine Dihydrochloride, (NH2)2 2HCl104.97
the USA by www.nextteq.com.]
Hydroquinone, C6H4(OH)2110.11 [123-31-9]Fine,
colorless or white, needle crystals. Darkens on exposure to
50 mL of water. Add carefully while stirring, 1 g of sodium
air and light. Soluble in water, in alcohol, and in ether.
bicarbonate. [CAUTIONThere may be a rapid evolution of
AssayAccurately weigh about 250 mg, and dissolve in a
carbon dioxide.] Titrate with 0.1 N iodine solution, deter-
mixture of 100 mL of water and 10 mL of 0.1 N sulfuric acid
mining the endpoint potentiometrically. Perform a blank de-
in a 250-mL conical flask. Add 3 drops of a 1 in 100 solu-
termination, and make any necessary corrections. Each mL
tion of diphenylamine in sulfuric acid, and titrate with 0.1 N
of 0.1 N iodine solution is equivalent to 2.63 mg of (NH2)2
ceric sulfate VS until the solution is red-violet in color. Each
2HCl. Not less than 98% is found.
mL of 0.1 N ceric sulfate is equivalent to 5.506 mg of
Hydrazine Hydrate, 85% in Water, (NH2)2 H2O50.06
C6H4(OH)2. Not less than 99% is found.
AssayTransfer 600 mg, accurately weighed, to a
Hydroxy Naphthol Blue (1-(2-Naphtholazo-3,6-disulfonic
100-mL volumetric flask. Dilute with water to volume, and
Acid)-2-naphthol-4-sulfonic Acid, Disodium Salt),
mix. Pipet 10 mL into a suitable beaker, and add 1.0 g of
C20H12N2O11S3Na2598.50 [165660-27-5]Deposited on
sodium bicarbonate and 50.0 mL of 0.1 N iodine VS. Titrate
crystals of sodium chloride in the concentration of about
the excess iodine with 0.1 N sodium thiosulfate VS, using
1%. Use ACS reagent grade.
starch TS as the indicator. Perform a blank determination,
3-Hydroxyacetophenone, C8H8O2136.15
and make any necessary correction. Each mL of 0.1 N io-
[121-71-1]Light brown powder chips and chunks. Melts
dine is equivalent to 1.252 mg of (NH2)2 H2O. Not less
at about 96. Sparingly soluble in chloroform, yielding a
than 83% is found.
clear, light yellow solution.
Hydrazine Sulfate, (NH2)2 H2SO4130.12
[10034-93-2]Use ACS reagent grade. [CAUTIONGreat
care should be taken in handling hydrazine sulfate because
it is a suspected carcinogen.]
Hydrindantin (2,2-Dihydroxy-2,2-biindan-1,1,3,3-tet-
0.25-mm 30-m capillary column coated with G1; the de-
rone), C18H10O6322.27 [5103-42-4]Sparingly soluble
tector and the injection port temperatures are maintained at
in hot water; soluble in methoxyethanol. When heated
above 200, it becomes reddish brown.
programmed to rise 10 per minute to 280 and held at
that temperature for 10 minutes. The area of the main peak
Hydriodic Acid, HI127.91 [10034-85-2]Use ACS
is not less than 97% of the total peak area.
reagent grade (containing not less than 47.0% of HI).
4-Hydroxyacetophenone, HOC6H4COCH3136.15
[NOTEFor methoxy determination (see Methoxy Determi-
[99-93-4]Gray powder, melting at about 109.
nation 431), use hydriodic acid ACS reagent grade 55%.
p-Hydroxybenzoic Acid, C7H6O3138.12 [99-96-7]
White crystals.
AssayTransfer about 700 mg, accurately weighed, to a [94035-02-6]Use a suitable grade with a substitution de-
suitable container, and dissolve in 50 mL of acetone. Add gree between 0.40 and 1.50.
100 mL of water, mix, and titrate with 0.5 N sodium hy- 8-Hydroxyquinoline (Oxine), C9H7NO145.16
droxide VS, determining the endpoint potentiometrically. [148-24-3]Use ACS reagent grade 8-Quinolinol.
Perform a blank determination, and make any necessary Hypophosphorous Acid, 50 Percent (Hypophosphorous
correction. Each mL of 0.5 N sodium hydroxide is equiva- Acid), HPH2O266.00 [6303-21-5]A colorless to faintly
lent to 69.06 mg of C7H6O3: not less than 97% is found. yellow liquid. Miscible with water and with alcohol.
Melting range 741: over a range of 2 that includes AssayAccurately weigh about 4 mL, dilute with 25 mL
216. of water, add methyl red TS, and titrate with 1 N sodium
4-Hydroxybenzoic Acid Isopropyl Ester, hydroxide VS: each mL of 1 N sodium hydroxide is equiva-
HOC6H4COOCH(CH2)2180.18 [4191-73-5]Use a suit- lent to 66.00 mg of HPH2O2. Not less than 48% is found.
able grade.
ChlorideAdd 0.2 mL to a mixture of 10 mL of silver ni-
trate TS and 5 mL of nitric acid, and heat until brown fumes
are no longer evolved: any white, insoluble residue remain-
Melting range 741: between 84 and 87. ing is negligible.
2-Hydroxybenzyl Alcohol, C7H8O2124.14
PhosphateDilute 1 mL with water to 50 mL, render al-
[90-01-7]Off-white flakes. Very soluble in alcohol, in chlo-
kaline with ammonia TS, filter if a precipitate is formed, and
roform, and in ether; soluble in 15 parts water and in ben-
add to the filtrate 5 mL of magnesia mixture TS: not more
zene.
than a slight precipitate is formed within 5 minutes.
Sulfate (Reagent test, Method I)Dilute 1 mL with water
matograph (see Chromatography 621), equipped with a
to 50 mL: 20 mL of the solution shows not more than
0.2 mg of SO4.
Hypoxanthine, C5H4N4O136.11 [68-94-0]White
to yellowish-white powder. Soluble in 1 N sodium hydrox-
ide. Use a suitable grade.
IgG-Coated Red CellsRed cells coated with human im-
munoglobulin (IgG). The reagent must be obtained from
manufacturers or suppliers licensed by the Center for Bio-
C7H8O2 peak is not less than 99% of the total peak area.
logics Evaluation and Research, Food and Drug Administra-
Melting range 741: between 83 and 85. tion. The use of reagents from an unlicensed manufacturer
4-Hydroxybutane-1-sulfonic Acid (4-Hydroxy- or supplier may invalidate the results.
1-butanesulfonic Acid), C4H10O4S154.19 [26978-64-3] [NOTEThere are many manufacturers and suppliers of
Use a suitable grade with a content of NLT 95%. [NOTEA these reagents that are licensed by the Center for Biologics
suitable grade is available as catalog number RM-967-C50 Evaluation and Research, Food and Drug Administration.
from www.cydexpharma.com.] Some examples of licensed manufacturers or suppliers are
N-(2-Hydroxyethyl)piperazine-N-(2-ethanesulfonic the following: Gamma Biologics, Houston, TX; and Ortho
acid), C8H18N2O4S238.3 [7365-45-9]Use a suitable Diagnostics, Raritan, NJ.]
grade. Imidazole, C3H4N268.08 [288-32-4]White to light
Hydroxylamine Hydrochloride, NH2OH HCl69.49 yellow crystals. Freely soluble in water. Use ACS reagent
[5470-11-1]Use ACS reagent grade. grade.
10-Hydroxynorandrostenedione (10-Hydroxy- Indene, C9H8116.16 [95-13-6]Colorless liquid.
19-norandrost-4-ene-3,17-dione), C18H24O3288.38Use a
suitable grade.
4-(4-Hydroxyphenyl)-2-butanone, C10H12O2164.20
AssayInject an appropriate volume into a gas chromat- 0.25-mm 10-m capillary column coated with a 1-m layer
ograph (see Chromatography 621) equipped with a flame- of methylsilicone; the injection port temperature is main-
ionization detector, helium being used as the carrier gas. tained at 200; the detector temperature is maintained at
The following conditions have been found suitable: a 0.25- 300; the column temperature is maintained at 100 and
mm 30-m capillary column coated with a 1-m layer of programmed to rise 10 per minute to 250. The area of
phase G43; the injection port temperature is maintained at the indene peak is not less than 99% of the total peak area.
20.
IndicatorsSee separate subsection.
C10H12O2 peak is not less than 98.5% of the total peak area.
Indigo CarmineUse Indigotindisulfonate Sodium.
Melting range 741: between 81 and 87. Indole (2,3-Benzopyrrole), C8H7N117.14
3-Hydroxyphenyldimethylethyl Ammonium Chloride [120-72-9]Use a suitable grade.
[Dimethylethyl(3-hydroxyphenyl)ammonium Chloride]Use Indole-3-carboxylic Acid, C9H7NO2161.2
Edrophonium Chloride. [771-50-6]Use a suitable grade.
D--4-Hydroxyphenylglycine, C8H9NO3167.16 Inosine, C10H12N4O5268.23 [58-63-9]White, crys-
[22818-40-2]Shiny leaflets. Sparingly soluble in water, in talline powder.
alcohol, in acetone, in ether, in chloroform, in ethyl acetate,
Melting point 741: about 90.
in benzene, and in glacial acetic acid; soluble in alkalies and
Inositol (Hexahydroxycyclohexane), C6H6(OH)6180.16
in mineral acids; freely soluble in warm 20% v/v hydrochlo-
[87-89-8]Fine, white crystals or a white, crystalline pow-
ric acid.
der; stable in air. Its solutions are neutral to litmus. Optically
Melting range 741: between 220 and 247, with de- inactive. One g dissolves in 5.7 mL of water. Slightly soluble
composition. in alcohol; insoluble in ether and in chloroform. Store in
2-(4-Hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5-bi- well-closed containers.
1H-benzimidazole Trihydrochloride Pentahydrate
623.97 [23491-44-3]Dark yellow to tan powder with a
green cast. Use a suitable grade.
Hydroxypropyl--cyclodextrin (Hydroxypropylbetadex),
C42H70O35(C3H6O)x with x = 7 molar substitution
Melting range 741: between 223 and 226. Specific gravity 841: between 0.863 and 0.868.
Loss on drying 731Dry it at 105 for 4 hours: it loses Refractive index 831: between 1.3900 and 1.3920 at
not more than 0.5% of its weight. 20.
Isobutyl Alcohol (2-Methyl-1-propanol),
(CH3)2CHCH2OH74.12 [78-83-1]Use ACS reagent
Iobenguane Sulfate (m-Iodobenzylguanidine Hemisulfate
grade.
Salt), C8H10IN3
1
/2H2SO4324.1White powder. Freely sol-
4-Isobutylacetophenone, C12H16O176Pale yellow liq-
uble in methanol.
uid. Soluble in chloroform, in glycerols, in alcohols, in ether,
and in fatty oils; insoluble in water. Use a suitable grade.
the use of plates coated with chromatographic silica gel
mixture, a developing system consisting of a mixture of bu-
tyl alcohol, water, and acetic acid (60:25:15), and examined
N-Isobutylpiperidone, C9H17NO155.24Use a suitable
under short-wavelength UV light, not more than a single
grade.
impurity spot of not more than 0.5% is observed.
Isoflupredone Acetate (9--Fluoroprednisolone Acetate),
Iodic Acid, HIO3175.91 [7782-68-5]Use ACS rea-
C23H29FO6420.47Use Isoflupredone Acetate USP.
gent grade.
Isomaltotriose (-D-Glucosyl-(1-6)--D-glucosyl-(1-6)--D-
Iodine, I2253.81 [7553-56-2]Use ACS reagent
glucose), C18H32O16504.4 [3371-50-4]White lyophi-
grade.
lized powder. Use a suitable grade.
Iodine Monobromide, IBr206.81 [7789-33-5]
Isonicotinic Acid, C6H5NO2123.11 [52-22-1]Use a
Black, gray, or blue-purple crystals, crystalline needles, or
suitable grade.
crystalline chunks.
Isonicotinic Acid HydrazideUse Isoniazid.
AssayPlace about 100 mL of acetic acid in a 150-mL
IsooctaneSee 2,2,4-Trimethylpentane.
beaker. Separately dissolve 2 g of potassium iodide in a min-
Isopropyl Acetate, C5H10O2102.13 [108-21-4]Use
imum volume of water, add this solution to the acetic acid,
a suitable grade.
and mix. Transfer about 200 mg of Iodine Monobromide,
Isopropyl Alcohol (2-Propanol), (CH3)2CHOH60.10
accurately weighed, to the beaker containing the potassium
iodide and acetic acid mixture, and stir to dissolve. Titrate
[NOTEFor use in assays and tests involving UV spectro-
immediately with 0.1 N sodium thiosulfate VS, determining
photometry, use ACS reagent grade Isopropyl Alcohol Suita-
the endpoint potentiometrically (see Titrimetry 541). Per-
ble for Use in UV Spectrophotometry.]
Isopropyl Alcohol, Dehydrated [67-63-0]Use Isopropyl
tion. Each mL of 0.1 N sodium thiosulfate is equivalent to
Alcohol that previously has been dried by being shaken with
20.681 mg of IBr. Not less than 97.5% is found.
a suitable molecular sieve capable of adsorbing water, and
Iodine Monochloride, ICl162.36 [7790-99-0]Use
filtered.
ACS reagent grade.
Isopropyl EtherSee Diisopropyl Ether.
Iodoethane, C2H5I155.9 [75-03-6]Use a suitable
Isopropyl Iodide (2-Iodopropane), C3H7I169.99
grade.
p-Iodonitrotetrazolium Violet, (2-(4-Iodophenyl)-3-(4-ni-
Isopropyl Myristate, C17H34O2270.45 [110-27-0]
trophenyl)-5-phenyltetrazolium chloride), C19H13ClIN5O2
Use Isopropyl Myristate (NF monograph). For use as a sol-
505.70Light yellow powder.
vent in sterility test procedures, Isopropyl Myristate con-
AssayWhen tested by thin-layer chromatography, with forms to the following additional specification:
pH of water extractTransfer 100 mL to a 250-mL centri-
mixture and a developing system consisting of a mixture of
fuge bottle, add 10 mL of twice-distilled water, close the
amyl alcohol, formic acid, and water (8:1:1), sprayed with
bottle with a suitable closure, and shake vigorously for
0.1% sodium thiosulfate solution, and examined under
60 minutes. Centrifuge the mixture at 1800 rpm for 20 min-
short-wavelength UV light, a single spot is exhibited, with
utes, aspirate the upper (isopropyl myristate) layer, and de-
trace impurities.
termine the pH of the residual water layer: the pH is not less
Melting point 741: 240, with decomposition. than 6.5.
Ion-Exchange ResinAn intimate mixture of 4 parts of a Isopropyl Myristate not conforming to the test for pH of
strongly acidic cation-exchanger in the hydrogen form (pro- water extract may be rendered suitable for use in sterility
duced by sulfonation of a styrene-divinylbenzene copolymer, test procedures as follows:
representing 8 to 10% divinylbenzene) and 6 parts of a Using a 20-mm 20-cm glass column, add activated alu-
strongly basic anion-exchanger in the hydroxyl form (pro- mina, and tamp down to a height of 15 cm. Pass 500 mL of
duced by amination with trimethylamine of a chlorometh- the isopropyl myristate through the column, using a slight
ylated styrene-divinylbenzene copolymer, representing 3 to positive pressure to maintain an even flow, and use the elu-
5% divinylbenzene). ate collected directly in the sterility test procedure.
[NOTEA suitable resin is Amberlite MB-150, available Isopropyl Salicylate, C6H4OHCOOCH(CH3)2180.20
from Sigma-Aldrich, www.sigma-aldrich.com.] Colorless liquid.
Iron Wire, FeAt. Wt. 55.847Use a suitable grade.
Isoamyl AlcoholUse Amyl Alcohol.
Isobutyl Acetate, C6H12O2116.16 [110-19-0]Clear,
colorless liquid. Slightly soluble in water. Miscible with alco-
hol.
2-mm 1.8-m glass column packed with 7% phase G2 on
AssayInject an appropriate specimen into a suitable gas support S1A; the injection port temperature is maintained at
chromatograph (see Chromatography 621) equipped with 250; the detector temperature is maintained at 310; the
a flame-ionization detector, helium being used as the carrier column temperature is maintained at 50 and programmed
gas. The following conditions have been found suitable: a to rise 10 per minute to 250. The area of the main peak is
0.25-mm 30-m capillary column coated with G2. The in- not less than 97% of the total peak area.
jection port temperature is maintained at 130; the column Isopropylamine (2-Aminopropane), C3H7NH259.11
temperature is maintained at 30 and programmed to rise [75-31-0]Clear, colorless, flammable liquid. Miscible with
10 per minute to 180 and held there for 10 minutes. The water, with alcohol, and with ether.
detector temperature is maintained at 300. The area of the
AssayTransfer about 0.2 g, accurately weighed, to a
main peak is not less than 99% of the total peak area.
suitable container, add 50 mL of water, and mix. Titrate
with 0.1 N hydrochloric acid VS, using a mixture of bromo-
cresol green TS and methyl red TS (5:1) as indicator. Each
mL of 0.1 N hydrochloric acid is equivalent to 59.11 mg of acid and lead acetate TS (1:10). Remove, dry, and cut into
C3H9N. Not less than 98% is found. 15-mm 40-mm strips.
Lead Monoxide (Litharge), PbO223.20
[1317-36-8]Heavy, yellowish or reddish-yellow powder. In-
between 31 and 33.
soluble in water and in alcohol; soluble in acetic acid, in
diluted nitric acid, and in warm solutions of the fixed alkali
20.
hydroxides.
Isorhamnetin, C16H12O7316.27 [418-19-3]Yellow
AssayAccurately weigh about 300 mg, freshly ignited in
powder.
a muffle furnace at 600 50, and dissolve it by warming
Melting point: greater than 300.
with 10 mL of water and 1 mL of glacial acetic acid. Dilute
Isovaleric Acid (3-Methylbutanoic Acid, Isovalerianic Acid,
with 75 mL of water, heat to boiling, add 50.0 mL of 0.1 N
Isopropylacetic Acid), C5H10O2102.13 [503-74-2]Use a
potassium dichromate VS, and boil for 2 to 3 minutes. Cool,
suitable grade.
transfer to a 200-mL volumetric flask with the aid of water,
Kaempferol, C15H10O6286.24 [520-18-3]Light yel-
dilute with water to volume, mix, and allow to settle. With-
low to yellow powder. It is a clear, bright yellow solution in
draw 100.0 mL of the clear liquid, and transfer to a glass-
alcohol.
stoppered flask. Add 10 mL of diluted sulfuric acid and 1 g
Kerosene [8008-20-6]A mixture of hydrocarbons,
of potassium iodide, insert the stopper, mix gently, and al-
chiefly of the methane series. A clear, colorless liquid. Spe-
low to stand for 10 minutes. Then titrate the liberated io-
cific gravity: about 0.80. Distills between 180 and 300.
dine, representing the excess of dichromate, with 0.1 N so-
L DesignationsSee packings for high-pressure liquid
dium thiosulfate VS, adding 3 mL of starch TS as the
chromatography under Reagents, Chromatographic Columns.
endpoint is approached: each mL of 0.1 N potassium
Lactose, C12H22O11 H2O360.31 [64-42-3]Use ACS
dichromate is equivalent to 7.440 mg of PbO. Not less than
reagent grade.
98% is found.
Alpha Lactose Monohydrate (-D-Lactose Monohydrate),
Insoluble in acetic acidDissolve 2 g in 30 mL of dilute
C12H22O11 H2O360.31White powder. The -D-lactose
glacial acetic acid (1 in 2), boil gently for 5 minutes, filter,
content is less than 3%.
wash the residue with diluted acetic acid, and dry at 105
AssayInject an appropriate derivatized specimen into a
for 2 hours: the residue weighs not more than 10 mg
gas chromatograph (see Chromatography 621) equipped
(0.5%).
with a flame-ionization detector, helium being used as the
Substances not precipitated by hydrogen sulfideCom-
carrier gas. The following conditions have been found suita-
pletely precipitate the lead from the filtrate obtained in the
ble: a 0.25-mm 30-m capillary column coated with a
test for Insoluble in acetic acid by passing hydrogen sulfide
1-m layer of phase G2; the injection port temperature is
into it, filter, and wash the precipitate with 20 mL of water.
maintained at 250; the detector temperature is maintained
To one-half of the mixed filtrate and washings add 5 drops
at 280; the column temperature is maintained at 230 and
of sulfuric acid, evaporate to dryness, and ignite at
programmed to rise 4 per minute to 280. The area of the
800 25 for 15 minutes: the residue weighs not more than
C12H22O11 H2O peak is not less than 97% of the total peak
5 mg (0.5%).
area.
Beta Lactose (-D-Lactose), C12H22O11342.30White to Volatile substancesAccurately weigh about 5 g, and heat
faint yellow powder. The -D-lactose content is not more strongly in a covered porcelain crucible: it loses not more
than 35%. than 2.0% of its weight.
Lead Nitrate, Pb(NO3)2331.21 [10099-74-8]Use
AssayInject an appropriate derivatized specimen into a
ACS reagent grade.
gas chromatograph (see Chromatography 621) equipped
Lead Perchlorate, Pb(ClO4)2 3H2O460.15Use ACS
with a flame-ionization detector, helium being used as the
reagent grade.
Lead Tetraacetate, C8H12O8Pb443.38 [546-67-8]
ble: a 0.25-mm 30-m capillary column coated with a
Colorless to pinkish crystals. Soluble in hot glacial acetic
1-m layer of phase G43; the injection port temperature is
acid, in chloroform, in nitrobenzene, and in tetrachloroeth-
maintained at 250; the detector temperature is maintained
ane. Hydrolyzed by water, producing brown lead dioxide
at 250; the column temperature is maintained at 20 and
and acetic acid. Unstable in air.
C12H22O11 peak is not less than 99% of the total peak area. Melting range 741: between 175 and 180.
Lanthanum Alizarin Complexan MixtureUse a suitable Linalool (3,7-Dimethyl-1,6-octadien-3-ol), C10H18O
grade. 154.25 [78-70-6]Use a suitable grade.
Lanthanum Chloride, LaCl3 (67)H2O [10025-84-0] Linoleic Acid, C18H32O2280.4 [60-33-3]Clear, col-
This reagent is available in degrees of hydration ranging orless liquid. Use a suitable grade.
from 6 to 7 molecules of water. Use ACS reagent grade. -Lipoic Acid, C8H14O2S2206.3 [1077-28-7]Yellow
Lanthanum Nitrate Hexahydrate, La(NO3)3 6H2O powder. Use a suitable grade.
433.01 [10277-43-7]Use a suitable grade with a con- Liquid PetrolatumUse Mineral Oil.
tent of NLT 99.9%. Lithium Chloride, LiCl42.39 [7447-41-8]Use ACS
Lanthanum Oxide, La2O3325.82 [1312-81-8]An reagent grade.
almost white, amorphous powder, practically insoluble in Lithium Hydroxide, LiOH H2O41.96 [1310-65-2]
water. It dissolves in dilute solutions of mineral acids and Use ACS reagent grade.
absorbs atmospheric carbon dioxide. Use Atomic Absorption Lithium Metaborate, LiBO249.75 [13453-69-5]
Spectroscopy grade. Use ACS reagent grade.
Lauryl Dimethyl Amine Oxide (N,N-Dimethyldodecyl- Lithium Nitrate, LiNO368.95 [7790-69-4]Colorless
amine-N-oxide),C14H31NO [1643-20-5]229.41Use a suita- crystals. Use a suitable grade labeled to contain not less
ble grade. than 97.0%.
[NOTEA suitable grade is available from Fluka, catalog Lithium Perchlorate, LiClO4106.39 [7791-03-9]
number 40234, www.sigma-aldrich.com.] Use ACS reagent grade.
Lead Acetate, Pb(C2H3O2)2 3H2O379.33 Lithium Sulfate, Li2SO4 H2O127.96 [10377-48-7]
[6080-56-4]Use ACS reagent grade. Use ACS reagent grade.
Lead Acetate PaperImmerse filter paper weighing Lithocholic Acid, C24H40O3376.57 [434-13-9]
about 80 g per square meter in a mixture of diluted acetic White powder.
mixture, a developing system consisting of a mixture of tol- while still warm to tight containers, as the anhydrous salt is
uene, 1,4-dioxane, and acetic acid (15.2:4.2:0.6), and very hygroscopic.
sprayed with a mixture of sulfuric acid and methanol (1:1), Malachite Green GSee Brilliant Green.
heated at 110 for 20 minutes, and examined visually and Maleic Acid, C4H4O4116.07 [110-16-7]White, crys-
under long-wavelength UV light, a single spot is exhibited. talline powder. Soluble in 1.5 parts of water, in 2 parts of
alcohol, and in 12 parts of ether.
Litmus [1393-92-6]A blue pigment prepared from vari- AssayDissolve about 2 g, accurately weighed, in
ous species of Rocella DeCandolle, Lecanora Acharius, or 100 mL of water and titrate with 1 N sodium hydroxide VS,
other lichens (Fam. Parmeliaceae). using phenolphthalein TS as the indicator. Each mL of 1 N
sodium hydroxide is equivalent to 58.04 mg of C4H4O4: not
DescriptionCubes, masses, fragments, or granules, of an
less than 99% of C4H4O4, calculated on the dried basis, is
indigo blue or deep violet color. Has the combined odor of
found.
indigo and violets, and tinges the saliva a deep blue. The
indicator substances it contains are soluble in water and less Loss on dryingDry it in vacuum over phosphorus pent-
soluble or insoluble in alcohol. oxide for 2 hours: it loses not more than 1.5% of its weight.
AshIt yields not more than 60.0% of ash. Residue on ignition 281: not more than 0.1%.
Locust Bean GumA gum obtained from the ground en- Maltotriose, C18H32O16504.44 [1109-28-0]Use a
dosperms of Ceratonia siliqua Linn e Taub. (Fam. Legumi- suitable grade with a content of not less than 95%.
nosae). Use Locust Bean Gum, FCC. Manganese DioxideSee Manganese Dioxide, Activated.
L-Lysine (2,6-Diaminohexanoic Acid), C6H14N2O2146.19 Manganese Dioxide, Activated (Manganese (IV) Oxide,
[56-87-1]Crystalline needles or hexagonal plates. Soluble Activated), MnO286.94 [1313-13-9]Use a suitable
in water; very slightly soluble in alcohol; insoluble in ether. grade.
Melamine (2,4,6-Triamino-1,3,5-triazine), C3H6N6126.1
Specific rotation 781: between +25.5 and +26.0.
Test solution: 20 mg per mL, in dilute hydrochloric acid
2-Mercaptoethanol (-Mercaptoethanol), C2H6OS
(1 in 2).
Nitrogen content, Method I 461: between 18.88% and
Mercuric Acetate, Hg(C2H3O2)2318.68
19.44% of N is found, corresponding to not less than
98.5% of C6H14N2O2, the test specimen previously having
Mercuric Bromide, HgBr2360.40 [7789-47-1]Use
been dried at 105 for 2 hours.
ACS reagent grade.
Magnesium, Mg24.305 [7439-95-4]Silvery metal
Mercuric Chloride, HgCl2271.50 [7487-94-7]Use
in ribbon form. Reacts slowly with water at room tempera-
ACS reagent grade.
ture. Dissolves readily in dilute acids with the liberation of
Mercuric Iodide, Red, HgI2454.40 [7774-29-0]Use
hydrogen.
ACS reagent grade.
AssayTransfer 1 g, accurately weighed, to a 250-mL vol- Mercuric Nitrate, Hg(NO3)2 xH2O342.62
umetric flask, and dissolve in a mixture of 15 mL of hydro- [10045-94-0]Use ACS reagent grade. This reagent is avail-
chloric acid and 85 mL of water. When solution is complete, able as either the mono- or dihydrate.
dilute with water to volume, and mix. Pipet 25 mL of the Mercuric Oxide, Yellow, HgO216.59
dilution into a 400-mL beaker, dilute with water to 250 mL, [21908-53-2]Use ACS reagent grade.
add 20 mL of ammoniaammonium chloride TS and a few Mercuric Sulfate, HgSO4296.65 [7783-35-9]Use
mg of eriochrome black T trituration, and titrate with 0.1 M ACS reagent grade.
edetate disodium VS to a blue endpoint. Each mL of 0.1 M Mercuric Thiocyanate, Hg(SCN)2316.76
edetate disodium VS is equivalent to 2.430 mg of Mg. Not [592-85-8]White, crystalline powder. Very slightly soluble
less than 99% is found. in water; soluble in solutions of sodium chloride; slightly sol-
Magnesium Acetate, Mg(C2H3O2)2 4H2O214.45 uble in alcohol and in ether.
[142-72-3]Use ACS reagent grade. Mercurous Nitrate Dihydrate (Mercury (I) Nitrate
Magnesium Chloride, MgCl2 6H2O203.30 Dihydrate), Hg2(NO3)2 2H2O561.22Use ACS reagent
[7786-30-3]Use ACS reagent grade. grade.
Magnesium Nitrate, Mg(NO3)2 6H2O256.41 Mercury, HgAt. Wt. 200.59 [7439-97-6]Use ACS
[10377-60-3]Use ACS reagent grade. reagent grade.
Magnesium Oxide, MgO40.30 [1309-48-4]Use Mesityl Oxide, C6H10O98.14 [141-79-7]Colorless
ACS reagent grade. liquid.
Magnesium Oxide, ChromatographicUse a suitable
grade.
Magnesium Perchlorate, Anhydrous, Mg(ClO4)2
Magnesium Silicate, ActivatedUse a suitable grade.
Magnesium Silicate, ChromatographicExtremely
white, hard, powdered (60- to 100-mesh) magnesiasilica
gel. Suitable for use as an adsorbant in column chromatog-
column temperature is maintained at 50 and programmed
raphy.
to rise 10 per minute to 200. The area of the C6H10O peak
Magnesium Sulfate, MgSO4 7H2O246.48
is not less than 98% of the total peak area.
Magnesium Sulfate, Anhydrous, MgSO4120.37
Metanil Yellow (Acid Yellow 36;
[7487-88-9]Anhydrous Magnesium Sulfate may be pre-
3-(4-Anilinophenylazo)benzenesulfonic Acid Sodium Salt),
pared as follows. Place a suitable quantity of magnesium
C18H14N3NaO3S375.38 [587-98-4]Use a suitable grade
sulfate (see above), preferably powdered, in a shallow ves-
with a dye content of NLT 70%.
sel, and expose to a temperature of about 80 for several
Metaphenylenediamine Hydrochloride (Metaphenylene-
hours with occasional stirring. Then heat at 275 to 300
diamine Dihydrochloride), C6H4(NH2)2 2HCl181.06White
until the weight is practically constant. Transfer the product
or slightly reddish-white, crystalline powder. Easily soluble in
water. On exposure to light it acquires a reddish color.
Store it protected from light.
SolubilityA solution of 1 g in 200 mL of water is color-
less.
Residue on ignition (Reagent test)Ignite 1 g with 0.5 mL tion. Each mL of 0.1 N perchloric acid is equivalent to
of sulfuric acid: the residue weighs not more than 1 mg 15.12 mg of C8H9NO2. Not less than 99.0% is found.
(0.1%).
[NOTEMetaphenylenediamine hydrochloride solution can
Methyl Arachidate (Eicosanoic acid, methyl ester),
be decolorized by treatment with a small quantity of acti-
C21H42O2326.56 [1120-28-1]Off-white flakes.
vated charcoal.]
Metaphosphoric Acid (Vitreous Sodium Acid Metaphos-
phate), HPO379.98 [37267-86-0]Use ACS reagent
thermal-conductivity detector, helium being used as the car-
grade.
Methacrylic Acid [79-41-4]Use a suitable grade.
a 2.0-mm 1.8-m glass column packed with 5% G2 phase
Methanesulfonic Acid, CH4O3S96.11 [75-75-2]
on support S1A; the injection port temperature is main-
Methanol (Methyl Alcohol), CH3OH32.04
Methanol, Aldehyde-Free, CH3OH32.04Dissolve
25 g of iodine in 1 L of methanol and pour the solution,
with constant stirring, into 400 mL of 1 N sodium hydrox-
Methyl Behenate, C23H46O2354.61 [929-77-1]
ide. Add 150 mL of water, and allow to stand for 16 hours.
White powder.
Filter, and boil under a reflux condenser until the odor of
AssayInject an appropriate specimen into a gas chro- iodoform disappears. Distill the solution by fractional distilla-
matograph (see Chromatography 621) equipped with a tion. It contains not more than 0.001% of aldehydes and
thermal conductivity detector, helium being used as the car- ketones.
rier gas. The following conditions have been found suitable: Methanol, AnhydrousUse Methanol.
a 2.0-mm 1.8-m glass column packed with 5% G3 phase Methanol, SpectrophotometricUse ACS reagent grade
on support S1A; the injection port temperature is main- Methanol Suitable for Use in UV Spectrophotometry.
tained at 300; the detector temperature is maintained at Methenamine (Hexamethylenetetramine; Urotropine;
300; the initial temperature of the oven is 220, which is Uritone; Hexamine), C6H12N4140.19 [100-97-0]Use
held for 2 minutes, and then programmed to rise 3 per ACS reagent grade Hexamethylenetetramine.
minute to attain a final temperature of 270, which is held 7-Methoxycoumarin (Herniarin; Methyl Umbelliferyl
for 10 minutes. The area of the C23H46O2 peak is not less Ether), C10H8O3176.17 [531-59-9]Use a suitable grade
than 98% of the total peak area. with a content of not less than 98%.
Methoxyethanol (Ethylene Glycol Monomethyl Ether; Melting range 741: between 54 and 56.
2-Methoxyethanol), CH3OCH2CH2OH76.09 Methyl Benzenesulfonate, C7H8O3S172.20
[109-86-4]Use ACS reagent grade. [80-18-2]Use a suitable grade.
2-Methoxyethanol (Ethylene Glycol Monomethyl Ether; Methyl Caprate, C11H22O2186.29 [110-42-9]Col-
Methoxyethanol), CH3OCH2CH2OH76.09 [109-86-4] orless liquid.
See Methoxyethanol.
Add the following:
mm 30-m capillary column coated with a 1-m layer of L
5-Methoxy-1H-benzimidazole-2-thiol (5-Methoxy-
2-benzimidazolethiol), C8H8N2OS180.23 [37052-78-1]
Use a suitable grade with a content of NLT 99.0%.L USP36
5-Methoxy-2-methyl-3-indoleacetic Acid, C12H13NO3
219.24 [2882-15-7]Off-white powder.
Methyl Caprylate, C9H18O2158.24 [111-11-5]Col-
100-mL beaker. Add 30 mL of methanol and dissolve by stir-
orless liquid.
ring. Add 40 mL of water, and mix. Titrate with 0.1 N so-
dium hydroxide VS, determining the endpoint potentiomet-
rically. Perform a blank determination and make any
necessary correction. Each mL of 0.1 N sodium hydroxide is
equivalent to 21.92 mg of C12H13NO3. Not less than 98% is
found.
ceed 3.
Methoxyphenylacetic Acid (-Methoxyphenylacetic Acid),
Methyl Carbamate, C2H5NO275.07 [598-55-0]
Methyl Acetate, C3H6O274.08 [74-20-9]Colorless
White crystals. Freely soluble in water.
liquid. Soluble in water. Miscible with alcohol and with
ether.
Methyl Chloroform (Methylchloroform; 1,1,
1-Trichloroethane), CH3CCl3133.40 [71-55-6]Use ACS
reagent grade.
20.
Methyl Erucate, C23H44O2352.59 [1120-34-9]Col-
Boiling range (Reagent test)Not less than 95% distills orless liquid.
between 57 and 58. Methyl Ethyl Ketone, CH3COC2H572.11 [78-93-3]
Methyl 4-Aminobenzoate, C8H9NO2151.16 Use ACS reagent grade 2-butanone.
[619-45-4]Off-white powder. Methyl Green (Methyl Green Zinc Chloride Double Salt;
Ethyl Green Zinc Chloride Double Salt; C.I. 42590),
C27H35Cl2N3 ZnCl2608.78 [7114-03-6]Use a suitable
50 mL of glacial acetic acid. Titrate with 0.1 N perchloric
grade for microscopy.
acid VS, determining the endpoint potentiometrically. Per-
Methyl Heptadecanoate, C18H36O2284.48
[1731-92-6]White, crystalline flakes.
AssayInject an appropriate specimen into a gas chro- AssayInject an appropriate volume into a gas chromat-
matograph (see Chromatography 621) equipped with a ograph (see Chromatography 621) equipped with a flame-
flame-ionization detector, helium being used as the carrier ionization detector, helium being used as the carrier gas.
gas. The following conditions have been found suitable: a The following conditions have been found suitable: a 0.25-
0.25-mm 30-m capillary column coated with a 1-m layer mm 30-m capillary column coated with a 1-m layer of
of phase G8; the injection port temperature is maintained at phase G2; the injection port temperature is maintained at
220; the detector temperature is maintained at 220; the 300; the detector temperature is maintained at 300; the
column temperature is maintained at 180 and pro- column temperature is maintained at 200 and pro-
grammed to rise 4 per minute to 220. The area of the grammed to rise 10 per minute to 300. The area of the
C18H36O2 peak is not less than 99% of the total peak area. C15H30O2 peak is not less than 99% of the total peak area.
Melting range 741: between 31 and 32. Refractive index 831: between 1.434 and 1.438 at 20.
Methyl Iodide, (Iodomethane), CH3I141.94 Methyl Oleate, C19H36O2296.49 [112-62-9]Color-
[74-88-4]Colorless, heavy, transparent liquid. Slightly solu- less liquid.
ble in water. Miscible with alcohol, with ether, and with
solvent hexane. Turns brown on exposure to light as a result
of liberation of iodine.
Use a suitable grade with a content of not less than 99%.
Methyl Isobutyl KetoneSee 4-Methyl-2-pentanone.
capillary column coated with a 1-m layer of phase G2; the
Methyl Laurate, C13H26O2214.34 [110-82-0]Color-
injection port temperature is maintained at 300; the detec-
less liquid.
tor temperature is maintained at 300; the column tempera-
AssayInject an appropriate volume into a gas chromat- ture is maintained at 230 and programmed to rise 10 per
ograph (see Chromatography 621) equipped with a flame- minute to 280. The area of the C19H36O2 peak is not less
ionization detector, helium being used as the carrier gas. than 99% of the total peak area.
Methyl Palmitate, C17H34O2270.45 [112-39-0]
White solid.
C13H26O2 peak is not less than 99.45% of the total peak
area.
Methyl Lignocerate, C25H50O2382.66 [2442-49-1]
White crystals.
Methyl Linoleate, C19H34O2294.47 [112-63-0]Col-
orless liquid.
Methyl Red (2-[4-Dimethylaminophenylazo]benzoic Acid;
C. I. Acid Red 2) C15H15N3O2, free acid 269.30
[493-52-7] C15H14N3O2Na, sodium salt291.28
[845-10-3]Use ACS reagent grade. The free acid is recom-
mended for nonaqueous titrations, particularly when an
aprotic solvent is used. The sodium salt is recommended for
titrations in aqueous media and also for nonaqueous titra-
tions where the medium is an amphiprotic solvent. The hy-
drochloride salt is recommended for titrations in aqueous
Methyl Linolenate, C19H32O2292.46 [301-00-8]
media and amphiprotic solvents.
Colorless liquid.
Methyl Stearate, C19H38O2298.50 [112-61-8]Off-
AssayInject an appropriate volume into a gas chromat- white, crystalline solid.
phase 14% cyanopropylphenyl-86% dimethylpolysiloxane;
the injection port temperature is maintained at 280; the
injection port temperature is maintained at 300; the detec-
detector temperature is maintained at 300; the column
tor temperature is maintained at 300; the column tempera-
ture is maintained at 200 and programmed to rise 10 per
10 per minute to 280. The area of the C19H32O2 peak is
minute to 300. The area of the C19H38O2 peak is not less
not less than 99% of the total peak area.
Methyl Methacrylate [80-62-6]Use a suitable grade.
Methyl Sulfoxide [67-68-5]See Dimethyl Sulfoxide.
Methyl Myristate, C15H30O2242.40 [124-10-7]Col-
Methyl Yellow (p-Dimethylaminoazobenzene), C14H15N3
orless liquid.
Methylamine, 40 Percent in Water, CH5N31.06 Melting range 741: between 59 and 63, but the
[74-89-5]Colorless liquid. range between beginning and end of melting does not ex-
ceed 2.
AssayUsing a syringe, transfer about 0.5 mL of a well-
4-Methylpentan-2-ol, C6H14O102.2Use a suitable
shaken specimen to 100 mL of water at a point below the
grade.
surface of the water. Determine the weight of the specimen
2-Methylpentane (2-Methyl-pentane; 1,1-Dimethylbutane;
by weighing the syringe before and after the transfer. Mix,
Isohexane), C6H1486.18 [107-83-5]Use a suitable
and titrate with 0.5 N hydrochloric acid VS, determining the
endpoint potentiometrically, using a silver-silver chloride pH
4-Methyl-2-pentanone (Methyl Isobutyl Ketone),
electrode and a calomel reference electrode. Perform a
(CH3)2CHCH2COCH3100.16 [108-10-1]Use ACS rea-
blank determination, and make any necessary correction.
gent grade.
Each mL of 0.5 N hydrochloric acid is equivalent to
2-Methyl-2-propyl-1,3-propanediol, C7H16O2132.20
15.53 mg of CH5N: between 39.0% and 41.0% is found.
[78-26-2]White crystals, melting at about 58.
N-Methylpyrrolidine (1-Methylpyrrolidine), C4H8NCH3
20.
p-Methylaminophenol Sulfate, (p-CH3NHC6H4OH)2
Mineral AcidUse Hydrochloric Acid or Sulfuric Acid.
H2SO4344.38 [55-55-0]Use ACS reagent grade.
5,800, 23,700, and 100,000 Molecular Weight (MW)
4-Methylbenzophenone, C14H12O196.25
Pullulan Standards (a commercial Pullulan Standard set
contains standards having several molecular weights:
Melting range 741: between 56.5 and 57.
5,800; 12,000; 24,000; 48,000; 100,000; 186,000;
Methylbenzothiazolone Hydrazone Hydrochloride,
380,000; and 750,000) [9057-02-7]Use a suitable grade.
C8H10ClN3S H2O233.7 [38894-11-0] (monohydrate
Each individual Pullulan Standard with a different molecular
form)215.70 (anhydrous form); [149022-15-1] (hydrate
weight, such as 5,800, 24,000, or 100,000, is equivalently
form)An almost white or yellowish, crystalline powder.
used.
Suitability for determination of aldehydesTo 2 mL of alde- [NOTEThe standard set is available from Polymer Labora-
hyde-free methanol add 60 L of a 1 g per L solution of tories (www.polymerlabs.com), Sigma-Aldrich (www.sigma-
propionaldehyde in aldehyde-free methanol and 5 mL of a aldrich.com), and Waters (www.waters.com).]
4 g per L solution of methylbenzothiazolone hydrazone hy- Molybdic Acid (85 Percent Molybdic Acid) [7782-91-4]
drochloride. Mix, and allow to stand for 30 minutes. Prepare Use ACS reagent grade.
a blank, omitting the propionaldehyde solution. Add Monobasic Potassium PhosphateSee Potassium Phos-
25.0 mL of a 2 g per L solution of ferric chloride to the test phate, Monobasic.
solution and to the blank, dilute with acetone to 100.0 mL, Monobasic Sodium PhosphateSee Sodium Phosphate,
and mix. The absorbance of the test solution, measured at Monobasic.
660 nm using the blank as compensation liquid, is not less Monochloroacetic Acid (Chloroacetic Acid, Chloroethanoic
than 0.62. Acid), CH2ClCOOH94.50 [79-11-8]Use ACS reagent
Use a suitable grade with a content of not less than 97%. grade.
(R)-(+)-alpha-Methylbenzyl Isocyanate ((R)-(+)- Monoethanolamine, (2-Aminoethanol) C2H7NO61.08
1-Phenylethyl Isocyanate), C9H9NO147.17 Use ACS reagent grade.
[33375-06-3]Use a suitable grade with a content of NLT Morin (Morin Hydrate; 2,3,4,5,7-Pentahydroxyflavone
99.0%. Monohydrate), C15H10O7 H2O320.25 [480-16-0]Use a
(S)-()--Methylbenzyl Isocyanate C9H9NO147.18 suitable grade.
[14649-03-7]Use a suitable grade. Morpholine (Tetrahydro-1,4-oxazine), C4H9NO87.12
Methylene Blue, C16H18ClN3S 3H2O373.90 [110-91-8]Use ACS reagent grade.
[7220-79-3]Dark green crystals or a crystalline powder, Naphthalene, C10H8128.17 [91-20-3]Monoclinic
having a bronzelike luster. One g dissolves in about 25 mL prismatic plates, or white scales or powder. Use a suitable
of water and in about 65 mL of alcohol. Soluble in chloro- grade with a content of not less than 98%.
form. Use a suitable grade with a dye content of not less 1,3-Naphthalenediol (Naphthoresorcinol), C10H6(OH)2
than 85%. 160.17 [132-86-5]Grayish-white to tan crystals or pow-
Methylene Chloride (Dichloromethane), CH2Cl284.93 der. Freely soluble in methanol; sparingly soluble in water,
[75-09-2]Use ACS reagent grade Dichloromethane. in alcohol, and in ether.
5,5-Methylenedisalicylic Acid (3,3-Methylene-bis[6-hy-
droxybenzoic Acid]), C15H12O6288.25 [122-25-8]Use a
Solubility in methanolDissolve 500 mg in 50 mL of
suitable grade.
methanol: the solution is clear and complete.
3-O-Methylestrone, C19H24O2284.39Use a suitable
2,7-Naphthalenediol (2,7-Dihydroxynaphthalene),
grade.
C10H8O2160.17 [582-17-2]Off-white to yellow, crys-
[NOTECommercially available as catalog number 1883-5
talline solid or powder. Dissolves in acetone.
from Research Plus, Inc., P.O. Box 324, Bayonne, NJ 07002,
fax number 908-754-2901, Web site: www.researchplus.
2-Naphthalenesulfonic Acid, C10H8O3S H2O226.25
com.]
[120-18-3]Off-white to light gray crystals. Soluble in
2-Methyl-5-nitroimidazole, C4H4N3O2127.10
water.
[NOTEAvailable as Catalog No. 13,625-5 from Sigma-Al- AssayDissolve about 1 g, accurately weighed, in
drich, www.sigma-aldrich.com.] 100 mL of water, add phenolphthalein TS, and titrate with
N-Methyl-N-nitroso-p-toluenesulfonamide (p-Tolyl- 0.1 N sodium hydroxide VS. Perform a blank determination,
sulfonylmethylnitrosamide), C8H10N2O3S214.24Light yel- and make any necessary correction. Each mL of 0.1 N so-
low crystals or powder. Insoluble in water; soluble in ben- dium hydroxide is equivalent to 22.63 mg of C10H8O3S
zene, in carbon tetrachloride, and in chloroform. H2O. Not less than 98.0% is found.
ceed 2.
1-Naphthol (Alphanaphthol), C10H7OH144.17 72240, available from Fluka Chemical Corp., fax
[90-15-3]Colorless or slightly pinkish crystals or crystalline 1-800-962-9591, Web site: www.sigma-aldrich.com.]
powder. Nickel Sulfate, NiSO4 6H2O262.85 [7786-81-4]
2-Naphthol (Betanaphthol), C10H7OH144.17 Use ACS reagent grade.
[135-19-3]White leaflets or crystalline powder. Discolors Nickel (II) Sulfate Heptahydrate, NiSO4 7H2O280.9
on exposure to light. Very slightly soluble in water; soluble [10101-98-1]Use a suitable grade.
in alcohol, in ether, in chloroform, and in solutions of alkali -Nicotinamide Adenine Dinucleotide, C21H27N7O14P2
hydroxides. 663.4 [53-84-9]White, very hygroscopic powder. Freely
soluble in water.
AssayDissolve 17.9 g of anhydrous dibasic sodium
Solubility in alcoholA solution of 1 g in 10 mL of alcohol
phosphate in water to make 500 mL (Solution A). Dissolve
is complete and colorless or practically so.
6.8 g of monobasic potassium phosphate in water to make
500 mL (Solution B). To a volume of Solution A, add Solution
0.05%.
B until the mixture is adjusted to a pH of 7.0 (about 2:1 by
AcidityShake 1 g with 50 mL of water occasionally dur-
volume of Solutions A and B) to obtain a pH 7.0 Buffer.
ing 15 minutes, and filter: the filtrate is neutral to litmus.
Transfer about 25 mg of -nicotinamide adenine dinucleo-
1-NaphtholBoil 100 mg with 10 mL of water until dis- tide, accurately weighed, to a 25-mL volumetric flask, dis-
solved, cool, and filter. Add to the filtrate 0.3 mL of 1 N solve in and dilute with water to volume, and mix. Transfer
sodium hydroxide and 0.3 mL of 0.1 N iodine: no violet 0.2 mL of this solution to a 10-mL volumetric flask, dilute
color is produced. with pH 7.0 Buffer to volume, and mix. Use this solution as
the Assay preparation. Determine the absorbances of the As- Insoluble in ammonia (naphthalene, etc.)Shake 500 mg
say preparation and the pH 7.0 Buffer in 1-cm cells at a with 30 mL of ammonia TS: the 2-naphthol dissolves com-
wavelength of 260 nm, using water as the reference. Calcu- pletely and the solution is not darker than pale yellow.
late the quantity, in mg, of C21H27N7O14P2 in the portion of Naphthol Dipotassium Disulfonate (2-Naphthol-6,8-di-
-nicotinamide adenine dinucleotide taken by the formula: potassium Disulfonate), C10H6K2O7S2380.48
(0.6634/17.6)(10/0.2)(25)(AA AB)
[NOTEA suitable grade is available as 2-naphthyl-6,8-
disulfonic acid dipotassium salt from Pfaltz and Bauer, Inc.,
in which AA and AB are the absorbances of the Assay prepa-
www.pfaltzandbauer.com.]
ration and the pH 7.0 Buffer, respectively. Not less than
Naphthol Disodium Disulfonate (2-Naphthol-3,6-diso-
94.5% is found.
dium Disulfonate), C10H6Na2O7S2348.26Use a suitable
Nicotinamide Adenine Dinucleotide Phosphate-
grade.
adenosine-5-triphosphate MixtureUse a suitable grade.
p-Naphtholbenzein, C27H18O2374.43 [145-50-6]
Red-brown powder. Use a suitable grade. SuitabilityWhen used in the assay of lactulose, deter-
-Naphthoquinone-4-sodium Sulfonate, C10H5NaO5S mine that a suitable absorbance-versus-concentration slope
260.20Yellow to orange-yellow crystals or crystalline pow- is obtained, using USP Lactulose RS , the reagent blank ab-
der. Soluble in about 10 parts of water; insoluble in alcohol. sorbance being not more than 0.020. The commercially
available reagent contains 64 mg of nicotinamide adenine
Loss on drying 731Dry it in vacuum at about 50: it
dinucleotide phosphate and 160 mg of adenosine-5-
triphosphate per vial. The mixture is buffered and stabilized.
Residue on ignition (Reagent test)Ignite 1 g of dried
For use in the Assay of lactulose it is diluted with water to
sample with 3 mL of sulfuric acid: the residue weighs be-
100 mL.
tween 265 and 280 mg (between 26.5% and 28.0%).
Nicotinic AcidUse Niacin (USP monograph).
Naphthoresorcinol (1,3-Dihydroresorcinol), C10H8O2
Ninhydrin, C9H4O3 H2O178.14 [485-47-2]Use
ACS reagent grade.
2-Naphthyl Chloroformate (Chloroformic Acid 2-Naphthyl
Nitric Acid, HNO363.01 [7697-37-2]Use ACS rea-
Ester), ClCOOC10H7206.62 [7693-50-7]Use a suitable
gent grade.
grade.
Nitric Acid, Diluted (10 percent HNO3) [7697-37-2]Di-
lute 143 mL of nitric acid with water to 1000 mL.
Nitric Acid, Fuming (90 Percent Nitric Acid), HNO3
1-Naphthylamine, C10H9N143.19 [134-32-7]Use
63.01 [7697-37-2]Use ACS reagent grade Nitric Acid,
a suitable grade.
90 Percent.
1-Naphthylamine Hydrochloride, C10H7NH2 HCl
Nitric Acid, Lead-FreeUse ACS reagent grade.
179.65 [552-46-5]White, crystalline powder that turns
LeadTo 100 g add 0.1 g of anhydrous sodium carbon-
bluish upon exposure to light and air. Soluble in water, in
ate and evaporate to dryness. Dissolve the residue in water,
alcohol, and in ether.
heating slightly, and dilute with the same solvent to
A 1 in 100 solution, make slightly acid with acetic acid,
50.0 mL. Determine the lead content by atomic absorption
gives a violet color with 5 drops of ferric chloride TS. A 1 in
spectrophotometry (see Spectrophotometry and Light-Scatter-
40 solution in diluted acetic acid is colorless and not more
ing 851) measuring the absorbance at 283.3 nm or
than slightly opalescent.
217.0 nm using a lead hollow-cathode lamp and an
Residue on ignition (Reagent test)Ignite 200 mg with a
airacetylene flame. It contains not more than 0.1 ppm of
few drops of sulfuric acid: the weight of the residue is negli-
lead (Pb).
gible.
Nitric OxideNitrogen Dioxide Detector TubeA fuse-
N-(1-Naphthyl)ethylenediamine Dihydrochloride,
sealed glass tube so designed that gas may be passed
C10H7NH(CH2)2NH2 2HCl259.17 [1465-25-4]Use
through it and containing suitable absorbing filters and sup-
ACS reagent grade.
port media for an oxidizing layer and the indicator diphenyl
Neutralized AlcoholSee Alcohol, Neutralized.
benzidine.
Nickel, Ni58.6934 [7440-02-0]Use a suitable
Measuring range: 0.5 to 10 ppm.
grade.
Nickel-Aluminum CatalystUse a suitable grade.
[NOTEA suitable grade is Raney Nickel, Active Catalyst,
available as aluminumnickel alloy, catalog number
Nitrilotriacetic Acid, N(CH2COOH)3191.14 sulfate, and dilute with water to 100 mL: the solution is
[139-13-9]Use ACS reagent grade. deep red in color and exhibits an absorption maximum at
4-Nitroacetophenone (p-Nitroacetophenone), 510 nm. To 1.0 mL of the solution add 1.0 mL of 0.01 M
C8H7NO3165.15 [100-19-6]Yellow crystals. ceric sulfate: the red color is discharged.
Nitroso R Salt (1-Nitroso-2-naphthol-3,6-disodium
AssayInject an appropriate ether solution of the speci-
Disulfonate), NOC10H4OH(SO3Na)2377.26 [525-05-3]
men (about 0.5 L) into a suitable gas chromatograph (see
Yellow crystals or crystalline powder. One g dissolves in
Chromatography 621) equipped with a thermal conductiv-
about 40 mL of water; insoluble in alcohol.
ity detector, helium being used as the carrier gas. The fol-
lowing conditions have been found suitable: a 4-mm SensitivenessDissolve 500 mg of sodium acetate in a so-
1.8-m stainless steel column containing 10% phase G1 on lution of 0.4 mg of cobaltous chloride (0.1 mg of cobalt) in
support S1A; the injection port and detector temperatures 5 mL of water. Add 1 mL of diluted acetic acid, and follow
are maintained at 200 and 300, respectively; the column with 1 mL of a solution of the nitroso R salt (1 in 500): a
temperature is maintained at 170 and programmed to rise red color, which is produced at once, persists when the so-
3 per minute to 220. The area of the 4-nitroacetophe- lution is boiled with 1 mL of hydrochloric acid for 1 minute.
none peak is not less than 97% of the total peak area. 1-Nitroso-2-naphthol, C10H7NO2173.17
[131-91-9]Brown to yellowish-brown powder. Insoluble in
water; soluble in alcohol, in benzene, in ether, in carbon
o-Nitroaniline, NO2C6H4NH2138.12 [88-74-4]Or-
tetrachloride, and in acetic acid.
ange-yellow crystals. Slightly soluble in cold water; soluble
in hot water; freely soluble in alcohol and in chloroform. It AssayTransfer about 250 mg, previously dried over silica
forms water-soluble salts with mineral acids. gel to constant weight and accurately weighed, to a glass-
stoppered flask, and dissolve in 10 mL of sodium hydroxide
solution (1 in 10). Cool the solution in an ice bath, add
p-Nitroaniline, NO2C6H4NH2138.12 [100-01-6]
dilute sulfuric acid (1 in 6) until a slight, permanent precipi-
Bright yellow, crystalline powder. Insoluble in water; soluble
tate is formed and the solution is slightly acid, then add 3 g
in alcohol and in ether.
of potassium iodide, shake to dissolve, add 20 mL of dilute
sulfuric acid (1 in 6), immediately insert the stopper in the
SolubilitySeparate 1-g portions dissolve in 30 mL of al-
flask, and allow to stand in the dark for 2 hours. Titrate the
cohol and in 40 mL of ether, respectively, to yield solutions
liberated iodine with 0.1 N sodium thiosulfate VS, adding
that are clear or practically so.
3 mL of starch TS as the endpoint is approached. Perform a
Residue on ignition (Reagent test): not more than 0.2%. complete blank determination, and make any necessary cor-
Nitrobenzene, C6H5NO2123.11 [98-95-3]Use ACS rection. Each mL of 0.1 N sodium thiosulfate is equivalent
reagent grade. to 8.66 mg of C10H7NO2: not less than 95.0% is found.
p-Nitrobenzenediazonium Tetrafluoroborate,
NO2C6H4N2BF4236.92 [456-27-9]Yellow-gold crystals.
Soluble in acetonitrile. [CAUTIONShock-sensitive; keep
Nitrous Oxide Certified Standard [10024-97-2]A con-
refrigerated.]
tainer of 99.9% nitrous oxide. It is available from most sup-
pliers of specialty gases.
low-actinic, 100-mL volumetric flask. Dissolve in 0.01 N hy-
Nonadecane, C19H40268.52 [629-92-5]White solid.
drochloric acid, dilute with 0.01 N hydrochloric acid to vol-
ume, and mix. Using low-actinic glassware, dilute 2.0 mL of
the resulting solution with spectrophotometric grade metha-
a thermal conductivity detector, helium being used as the
nol to 50.0 mL. Measure the absorbance of this solution in a
1-cm cell at about 255 nm, using methanol as the blank.
ble: 3-mm 1.8-m stainless steel column containing 5%
Calculate the absorptivity of the solution by dividing the
phase G2 on support S1AB; the injection port temperature
measured absorbance by the concentration in g per mL.
is maintained at 330; the detector temperature is main-
Calculate the assay value by the formula:
tained at 300; and the oven temperature is held initially at
190 and allowed to rise gradually to 250. The area of the
100a/59.4
nonadecane peak is not less than 99% of the total peak
area.
in which a is the absorptivity of the solution: not less than
Melting range 741: between 31.5 and 33.5. 95.0% is found.
Nonanoic Acid, C9H18O2158.24 [112-05-0]Clear, p-Nitrobenzyl Bromide, NO2C6H4CH2Br216.03Al-
colorless to faint yellow liquid. Miscible with water and with most white to pale yellow crystals, darkening on exposure
methanol. to light. Practically insoluble in water; freely soluble in alco-
hol, in ether, and in glacial acetic acid. Store in tight, light- AssayAccurately weigh about 500 mg, transfer to a
resistant containers. suitable container, add 30 mL of water, and mix. Add 40 mL
of water, and mix. Add phenolphthalein TS, and titrate with Melting range 741: between 98 and 100.
SolubilitySeparate 200-mg portions yield clear solutions
in 5 mL of alcohol and in 5 mL of glacial acetic acid.
200 mg.
Nonionic Wetting AgentUse a suitable amphoteric
4-(p-Nitrobenzyl)pyridine, C12H10N2O2214.22
surfactant.
[1083-48-3]Yellow crystals. Soluble in acetone. Use a suit-
[NOTEA suitable grade is commercially available as Triton
able grade with a content of not less than 98%.
X-100 or Octoxynol 9.]
Nitromethane, CH3NO261.04 [75-52-5]Use ACS
Nonoxynol-9 (Igepal CO-630, Semicid, Staycept, Tergitol
reagent grade.
TP-9, Sterox), (C2H4O)nC15H24O [26027-38-3]Use a suita-
5-Nitro-1,10-phenanthroline, C12H7N3O2225.20
ble grade.
[4199-88-6]White powder. Soluble in water.
1-Nonyl Alcohol (1-Nonanol), CH3(CH2)8OH144.25
Suitability as redox indicatorDissolve 25 mg in a mini-
mum volume of diluted sulfuric acid, add 10 mg of ferrous
AssayNot less than 97% of C9H20O is found, a suitable [NOTEAvailable from Draeger Safety, Inc., www.draeger.
gas chromatograph equipped with a flame-ionization detec- com.]
tor and helium being used as the carrier gas at a flow rate Oligo-deoxythymidinePolymeric length: 18. Use a suit-
of about 40 mL per minute. The following conditions have able grade.
been found suitable: a 3.2-mm 1.83-m stainless steel col- [NOTEA suitable grade is available from BD Biosciences,
umn packed with 20% phase G16 on support S1A; the in- www.bdbiosciences.com.]
jection port, column, and detector temperatures are main- Orange G (the sodium salt of azobenzene-betanaphthol
tained at about 250, 160, and 310, respectively. disulfonic acid), C6H5N:NC10H4(OH)(SO3Na)2-2,6,8452.37
[1936-15-8]Orange to brick-red powder or dark red crys-
tals. Readily soluble in water, yielding an orange-yellow so-
20.
lution; slightly soluble in alcohol; insoluble in ether and in
n-Nonylamine (1-Aminononane), C9H21N143.27
chloroform. The addition of tannic acid TS to its 1 in 500
solution causes no precipitation (acid color). The addition of
Nonylphenol Polyoxyethylene Ether, (CH3)3CCH2C
hydrochloric acid to a mixture of 500 mg of zinc dust and
(CH3)2CH2C6H4O(CH2CH2O)XH, where x is approximately 40
10 mL of its 1 in 500 solution produces decolorization.
to 19002100 [9016-45-9]White solid; melts at approxi-
When filtered, the colorless filtrate, on standing exposed to
mately 44. Use a suitable grade.
air, does not regain its original color (presence of azo-group).
Nonylphenoxypoly(ethyleneoxy)ethanolClear, viscous,
When heated, orange G does not deflagrate (distinction
pale yellow liquid. May exhibit slight solidification on cool-
from nitro colors). The addition of barium or calcium chlo-
ing; warming with agitation will restore to original condi-
ride TS to a concentrated solution of orange G produces a
tion. Density: about 1.06. Soluble in alcohol, in xylene, and
colored, crystalline precipitate. The addition of hydrochloric
in water. Suitable for use in gasliquid chromatography.
acid to its 1 in 500 solution produces no change; the addi-
[NOTEA suitable grade is Igepal CO 710, available
tion of sodium hydroxide TS to a similar solution produces a
from General Aniline and Film Corp., 140 West 51st St.,
yellowish red to a Bordeaux color but no precipitation. Or-
New York, NY 10020.]
ange G dissolves in sulfuric acid with an orange to yellow-
Normal Butyl AlcoholSee Butyl Alcohol.
ish-red color. No change in color results upon diluting the
Normal Butyl NitriteSee n-Butyl Nitrite.
solution cautiously with water.
Normal ButylamineSee n-Butylamine.
Orcinol (5-Methylresorcinol), C7H8O2 H2O142.15
n-Octadecane, CH3(CH2)16CH3254.49 [593-45-3]
[6153-39-5]White to light tan crystals.
Use a gas chromatographic standard with a content of not
less than 99.5%. AssayTransfer about 60 mg, accurately weighed, to a
Octadecyl Silane [18623-11-5]This reagent is formed in 100-mL volumetric flask, dissolve in methanol, dilute with
situ by reaction of the column support with a suitable silylat- methanol to volume, and mix. Transfer 5.0 mL of this solu-
ing agent such as octadecyl trichlorosilane. tion to a 50-mL volumetric flask and dilute with methanol
Octanesulfonic Acid Sodium Salt (Sodium 1-octanesulfon- to volume, and mix. Using a suitable spectrophotometer,
ate), C8H17NaO3S216.27 [5324-84-5]Use a suitable 1-cm cells, and methanol as the blank, record the absorb-
grade with a content of not less than 99.0%. ance of the solution at the wavelength of maximum absorb-
1-Octanol (Alcohol C8; Capryl Alcohol; Octyl Alcohol), ance at about 273 nm. From the observed absorbance, cal-
C8H18O130.23 [111-87-5]Use ACS reagent grade. culate the absorptivity (see Spectrophotometry and Light-
Octanophenone, C14H20O204.31 [1674-37-9]Col- scattering 851): the absorptivity is not less than 13.2, cor-
orless liquid. responding to not less than 98% of C7H8O2 H2O.
Assay Melting range 741: between 58 and 61.
OrthophenanthrolineSee 1,10-Phenanthroline.
MOBILE PHASEPrepare a filtered and degassed mixture of
Osmium Tetroxide (Osmic Acid; Perosmic Anhydride),
acetonitrile and water (7:3).
OsO4254.23 [20816-12-0]Use ACS reagent grade.
Oxalic Acid, H2C2O4 2H2O126.07 [6153-56-6]Use
ACS reagent grade.
254-nm detector and a 4.6-mm 15.0-cm column that
3,3-Oxydipropionitrile, O(CH2CH2CN)2124.14
contains packing L1. The flow rate is about 2 mL per min-
[1656-48-0]Clear, colorless to slightly yellow liquid. Re-
ute. The area of the C14H20O peak is not less than 99% of
fractive index: about 1.446 at 20.
Boiling range: between 174 and 176 at 10 mm of
mercury.
Octoxynol 9See Nonionic Wetting Agent.
Oxygen-Helium Certified StandardA mixture of 1.0%
(p-tert-Octylphenoxy) nonaethoxyethanol, C34H62O11
oxygen in industrial grade helium. It is available from most
646.85Use a suitable grade.
suppliers of specialty gases.
Packings for High-Pressure Liquid Chromatography
Tergitol Nonionic NPX, and as Triton N101, from rea-
See packings for high-pressure liquid chromatography in the
gent suppliers.]
Chromatographic Reagents section under Chromatography
(p-tert-Octylphenoxy)polyethoxyethanolUse a suitable
621.
grade.
Palladium CatalystUse a suitable grade.
[NOTEA suitable grade is available commercially as Tri-
[NOTEA suitable grade is available commercially as Pal-
ton 100 from reagent suppliers.]
ladium Catalyst, Type I (5% Palladium on Calcium Carbon-
Octyl Sulfate, Sodium Salt, C8H17O4SNa232.27White
ate), from Engelhard Industries, Inc., fax number (864)
powder.
885-1375.]
SolubilityA 2-g portion dissolves in 100 mL of water.
Palladium Chloride, PdCl2177.33 [7647-10-1]
Melting range 741: between 195 and 197, with Brown, crystalline powder. Soluble in water, in alcohol, in
decomposition. acetone, and in diluted hydrochloric acid.
Odorless Absorbent PaperSee Filter Paper, Quantitative.
AssayDissolve 80 mg, accurately weighed, in 10 mL of
Olefin Detector TubeA fuse-sealed glass tube so de-
diluted hydrochloric acid, dilute with water to 50 mL, and
signed that gas may be passed through it and containing
add 25 mL of a 1 in 100 solution of dimethylglyoxime in
alcohol. Allow to stand for 1 hour, and filter. Check for com-
tor in a stabilized form of permanganate.
plete precipitation with the dimethylglyoxime solution. Ig-
Measuring range: 0.06 to 3.2 Vol.% Propylene, 0.04 to nite the precipitate in a tared platinum crucible at 850 for
2.4 Vol.% Butylene. 2 hours, cool, and weigh the palladium. The weight of the
residue is not less than 59.0% of the weight of the test Add 5 mL of hydrochloric acid and 50 mL of water, and stir
specimen. until dissolved. Cool to about 15, add about 25 g of
Palladous ChlorideSee Palladium Chloride. crushed ice, and slowly titrate with 0.1 M sodium nitrite VS
Pancreatic Digest of Casein (a bacteriological peptone; until a glass rod dipped into the titrated solution produces
Tryptone)A grayish-yellow powder, having a characteristic, an immediate blue ring when touched to starch iodide pa-
but not putrescent, odor. Freely soluble in water; insoluble per. When the titration is complete, the endpoint is repro-
in alcohol and in ether. ducible after the mixture has been allowed to stand for
1 minute. Each mL of 0.1 M sodium nitrite is equivalent to
Nitrogen content (Reagent test)Determine by the
13.71 mg of C7H7NO2. Not less than 98.5% is found.
Kjeldahl method: 9.0%14.0% is found.
it loses not more than 7.0% of its weight. Loss on drying 731Dry it at 105 for 2 hours: it loses
Residue on ignition 281Ignite 500 mg with 1 mL of
sulfuric acid: the residue weighs not more than 75 mg Residue on ignition (Reagent test): not more than 0.1%.
(15%). Paraformaldehyde, (CH2O)n [30525-89-4]Fine, white
powder.
Microbial contentNMT 10,000 cfu/g.
AssayTransfer about 1 g, accurately weighed, to a
Bacteriological testPrepare medium of the following
250-mL conical flask containing 50.0 mL of 1 N sodium hy-
composition:
droxide VS, and mix by swirling. Immediately, and slowly,
2% of digest, 0.5% of sodium chloride, and 1.5% of agar
add 50 mL of hydrogen peroxide TS, previously neutralized
in purified water.
to bromothymol blue, through a small funnel placed in the
Adjust with diluted hydrochloric acid or diluted sodium
neck of the flask. After the reaction moderates, rinse the
hydroxide to a pH of 7.27.4. Autoclave at 121 for 15 min.
funnel and inner wall of the flask with water, allow the solu-
Growth-supporting propertiesSlants of the above me-
tion to stand for 30 minutes, add bromothymol blue TS,
dium, inoculated with Escherichia coli ATCC 25922, Enter-
and titrate the excess alkali with 1 N sulfuric acid VS. Each
obacter aerogenes ATCC 13048, Salmonella enterica ATCC
mL of 1 N sodium hydroxide is equivalent to 30.03 mg of
14028, Pseudomonas aeruginosa ATCC 27853, Staphylococ-
HCHO: not less than 95% is found.
cus aureus ATCC 25923, and Staphylococcus epidermidis
Residue on ignition: not more than 0.1%.
ATCC 12228, show characteristic growth after incubation
for 24 h. Solubility in ammoniaDissolve 5 g in 50 mL of ammonia
The above medium, to which 5% of sheep blood or TS: a practically clear, colorless solution results.
rabbit blood has been added, and which has been inocu-
ReactionShake 1 g with 20 mL of water for about
lated and poured into Petri dishes, shows characteristic al-
1 minute, and filter: the filtrate is neutral to litmus.
pha or beta zones around colonies of Streptococcus
Pectate Lyase [9015-75-2]An enzyme obtained from
pneumoniae ATCC 6305 and Streptococcus pyogenes ATCC
Aspergillus sp. Light brown, viscous liquid. Specific gravity is
49117, recognizable within 24 h and fully developed after
about 1.5. It is readily soluble in water. It is supplied at
48 h of incubation.
approximately 14 units per mL (at pH 8.0 in Tris-HCl buffer
The above medium, to which 10% of sheep blood or
[50 mM of Tris(hydroxymethyl)aminomethane containing
rabbit blood has been added, and which then has been
1 mM of CaCl2, pH 8.0] in a solution of 50% glycerol and
heated to 8090 until the blood has turned chocolate-
0.02% sodium azide. One unit is defined as the enzyme
brown, permits the growth of Neisseria gonorrhoeae ATCC
activity that produces 1 mol of unsaturated product per
19424 colonies within 48 h when incubated in an atmos-
minute.
phere containing 10% of carbon dioxide.
Activity
Pancreatin [8049-47-6]Use a grade of pancreatin which
PECTIN SOLUTIONTransfer a quantity of Pectin, equivalent
meets the USP requirements for amylase, lipase, and prote-
to 0.05 g on the dried basis, to a 100-mL volumetric flask.
ase activities specified for the official substance.
[NOTEPectin has a molecular weight of 103,000 Da; its de-
Papaic Digest of Soybean MealA soluble nutrient ma-
gree of esterification (percentage of galacturonic acid
terial prepared by the action of the enzyme papain on soy-
groups substituted with methyl) is 12.] Moisten with 0.1 mL
bean meal followed by suitable purification and concentra-
of 2-propanol. Add 50 mL of water to the flask, and mix the
tion. It contains fermentable carbohydrates.
solution with a magnetic stirrer. Use 0.5 N sodium hydrox-
ide to adjust the solution to a pH of 12. Stop the stirrer,
and allow the solution to stand undisturbed at room tem-
perature for 15 minutes. Adjust the solution with 0.5 N hy-
1 mL of sulfuric acid: the residue weighs not more than
drochloric acid to a pH of 8.0. Dilute with water to volume.
75 mg (15.0%).
TRIS BUFFER SOLUTIONTransfer 6.055 g of Tris(hydroxy-
Coagulable proteinHeat a filtered solution (1 in 20) to
methyl)aminomethane and 0.147 g of calcium chloride
boiling: no precipitate forms.
(CaCl2 H2O) to a 1000-mL volumetric flask containing
Microbial contentNMT 10
4
cfu/g 950 mL of water, and mix. Adjust the solution with 1 N hy-
drochloric acid to a pH of 8.0. Dilute with water to volume.
Kjeldahl method, using a test specimen previously dried at DILUTED PECTATE LYASETransfer 0.5 mL of Pectate Lyase to
105 to constant weight: not less than 8.5% is found. a 50-mL volumetric flask, dilute with Tris buffer solution to
Paper, Odorless AbsorbentSee Filter Paper, Quantita- volume, and mix.
tive.
PROCEDUREAdd the solutions set forth in the table below
Para-aminobenzoic Acid (p-Aminobenzoic Acid),
to quartz cuvettes.
H2NC6H4COOH137.14 [150-13-0]White or slightly
yellow crystals or crystalline powder, becoming discolored
Tris Diluted
on exposure to air or light. One g dissolves in 170 mL of
buffer Pectin pectate
water, in 9 mL of boiling water, in 8 mL of alcohol, and in
solution solution lyase Water
50 mL of ether. Freely soluble in solutions of alkali hydrox-
Label (mL) (mL) (mL) (mL)
ides and carbonates; soluble in warm glycerin; sparingly sol-
Enzyme blank 0.5 1.0 0 1.0
uble in diluted hydrochloric acid; slightly soluble in chloro-
Test blank 0.5 0 0.5 1.5 form. Store in tight, light-resistant containers.
Test solution 0.5 1.0 0.5 0.5
AssayAccurately weigh about 300 mg, previously dried
at 105 for 2 hours, and transfer to a beaker or casserole.
Perform the test on the solutions so obtained, using a suita- 2.0% HEMOGLOBIN SOLUTIONDissolve and dilute 2.5 g of
ble UV-Vis spectrophotometer (see Spectrophotometry and bovine hemoglobin with water to 100 mL. Dilute 80 mL of
Light-Scattering 851) and using water as the blank. Mix the this solution with 0.3 M hydrochloric acid to a volume of
solutions well at time 0, and immediately measure the ab- 100 mL.
sorbances at 235 nm. Record the value for the Enzyme
TRICHLOROACETIC ACID SOLUTIONDilute 5 g of trichloroace-
blank, A0EB ; for the Test blank, A0TB ; and for the Test solu-
tic acid with water to 100 mL.
tion, A0TS . After incubation at room temperature for
TEST SOLUTIONTransfer 5.0 mL of 2.0% Hemoglobin solu-
30 minutes, determine the absorbance again at 235 nm for
tion to a suitable container equilibrated at 37. Add 1.0 mL
the Enzyme blank, A30EB ; for the Test blank, A30TB ; and for
of Pepsin solution, mix by swirling, and incubate at 37 for
the Test solution, A30TS . One unit is defined as the enzy-
10 minutes. Immediately add 10.0 mL of Trichloroacetic acid
matic activity that produces 1 mol of unsaturated product
solution, mix by swirling, and incubate at 37 for 5 minutes.
from pectin per minute. Calculate the Pectate Lyase activity,
Pass through a filter having a 0.8-m or finer porosity.
in units per mL, using the following formula:
CONTROL SOLUTIONTransfer 5.0 mL of 2.0% Hemoglobin
solution to a suitable container equilibrated at 37. Mix by
50(10
3
)[(A30TS A30EB A30TB) (A0TS A0EB A0TB)]/
swirling, and incubate at 37 for 10 minutes. Immediately
30235L
add 10.0 mL of Trichloroacetic acid solution and 1.0 mL of
Pepsin solution, mix by swirling, and incubate at 37 for
in which 50 is the volume, in mL, of Diluted pectate lyase;
5 minutes. Pass through a filter having a 0.8-m or finer
10
3
is the unit conversion factor; 30 is the time, in minutes,
porosity.
of the reaction; 235 is the molar extinction coefficient, in
M
1
cm
1
, of the reaction product (4600 M
1
cm
1
); and L is PROCEDUREDetermine the absorbances of the Test solu-
the path length, in cm, of the reaction cuvette (1 cm). Alter- tion and Control solution, in 1-cm cells, at a wavelength of
natively, these solutions, after being mixed in the cuvettes, about 280 nm, using water as the reference. Calculate the
can be immediately measured at 235 nm continuously in a activity of the portion of Purified Pepsin taken by the
recording UV-Vis spectrophotometer set up for kinetic as- formula:
says. The result is obtained by correcting the blank determi-
10,000(AU AC) nation, using the Enzyme blank and the Test blank.
PenicillinaseSee Beta-lactamase.
in which AU and AC are the absorbances of the Test solution Pentadecane, C15H32212.41 [629-62-9]Colorless
and the Control solution, respectively. liquid.
Peptic Digest of Animal Tissue (a bacteriological pep-
tone)Tan powder, having a characteristic, but not putres-
cent, odor. Soluble in water; insoluble in alcohol and in
ether. An autoclaved solution (2 in 100) is clear and is neu-
tral or nearly so in its reaction.
Nitrogen contentDetermine by the Kjeldahl method, us- of phase G2; the injection port temperature is maintained at
ing a test specimen previously dried at 105 to constant 280; the detector temperature is maintained at 300; and
weight: between 9.0% and 14.0% of nitrogen is found. the column temperature is maintained at 180 and pro-
grammed to rise 10 per minute to 280. The area of the Residue on ignition 281Ignite 500 mg with 1 mL of
C15H32 peak is not less than 99% of the total peak area. sulfuric acid: the residue weighs not more than 75 mg
(15%). Refractive index 831: between 1.430 and 1.434 at
20. Loss on drying 731Dry it at 100 to constant weight:
Add the following:
4
cfu/g
L
Pentafluoropropanoic Acid (Pentafluoropropionic Acid),
C3HF5O2164.03 [422-64-0]Use a suitable grade with a Bacteriological test (for growth promotion and freedom
content of NLT 97%.L USP36 from fermentable carbohydrates)
Pentane (n-Pentane), C5H1272.15 [109-66-0]
Medium: 2% of digest and sufficient phenol red TS to
Clear, colorless, flammable liquid. Very slightly soluble in
give a perceptible color in water; adjust the pH to 7.27.4.
water. Miscible with alcohol, with ether, and with many or-
Place 10 mL of Medium in test tubes containing a Durham
ganic solvents. Specific gravity: about 0.62.
fermentation tube. Autoclave at 121 for 15 min. After
Boiling range (Reagent test)Not less than 95% distils autoclaving and standing for 24 h, the Medium is clear.
between 34 and 36. In separate tubes for each organism, inoculate 10 mL of
1-Pentanesulfonic Acid Sodium SaltSee Sodium Medium with Escherichia coli ATCC 25922 and Enterococcus
1-Pentanesulfonate. faecalis ATCC 12953. Incubate at 35 2 for 48 h.
2-Pentanone, C5H10O86.13 [107-87-9]Use a suita- It meets the following criteria for bacteria-nutrient proper-
ble grade. ties: Escherichia coli ATCC 25922 and Enterococcus faecalis
Pepsin [9001-75-6]Use Pepsin (Enzyme Preparations) ATCC 12953 growth evident, no acid or only a trace in the
FCC, having an activity of 1.0 to 1.17 Pepsin units per mg. inner tube, and no gas produced after incubation at 48 h.
Pepsin of higher activity may be reduced to this activity by Peptone, Dried (Meat Peptone)Reddish-yellow to
admixture with pepsin of lower activity or with lactose. brown powder, having a characteristic, but not putrescent,
Pepsin, PurifiedA white or yellowish-white powder, odor. Soluble in water, forming a yellowish-brown solution
spongy mass, or translucent scales or granules. Freely solu- having a slight acid reaction; insoluble in alcohol and in
ble in water, producing more or less opalescence; practically ether.
insoluble in alcohol, in chloroform, and in ether. Purified
Pepsin used in the second tier of the Dissolution test has an
Kjeldahl method, using a test specimen previously dried at
activity that is determined by the following method.
105 to constant weight: between 12% and 18% of nitro-
Activity gen is found.
PEPSIN SOLUTIONTransfer about 2.5 mg of Purified Pepsin, Residue on ignition (Reagent test)Ignite 500 mg with
accurately measured, to a 100-mL volumetric flask, dilute 1 mL of sulfuric acid: the residue weighs NMT 75 mg
with 10 mM hydrochloric acid to volume, and mix. [NOTE (15.0%).
Prepare immediately before use.]
Loss on drying 731Dry it at 105 to constant weight: 0.1 N sulfuric acid, representing this difference, is equivalent
it loses not more than 7.0% of its weight. to 11.91 mg of C6H5NCO: not less than 97.0% of C6H5NCO
is found.
2-Phenylacetamide (-Phenylacetamide), C8H9NO
135.16 [103-81-1]Bimorphous plates or leaflets.
4
cfu/g
Slightly soluble in water. Use a suitable grade.
Perchloric Acid (70 Percent Perchloric Acid), HClO4
100.46 [7601-90-3]Use ACS reagent grade (containing
dl-Phenylalanine, C9H11NO2165.19 [150-30-1]
between 69.0% and 72.0% of HClO4).
Periodic Acid, H5IO6227.94 [10450-60-9]White to
o-Phenylenediamine Dihydrochloride, C6H8N2 2HCl
pale yellow crystals. Very soluble in water. Undergoes slow
181.1White powder.
decomposition to iodic acid. Use ACS reagent grade.
Petroleum BenzinSee Hexane, Solvent. AssayWhen tested by thin-layer chromatography, with
Phases for Gas ChromatographySee phases for gas the use of plates coated with chromatographic silica gel
chromatography in the Chromatographic Columns section mixture and a developing system consisting of a mixture of
under Chromatography 621. butyl alcohol, water, and acetic acid (12:5:3), and examined
Phenacetin [62-44-2]Use a suitable grade. under short-wavelength UV light, a single spot is exhibited,
1,10-Phenanthroline (Orthophenanthroline), C12H8N2 with trace impurities.
H2O198.22 [5144-89-8]Use ACS reagent grade. p-Phenylenediamine DihydrochlorideSee p-Phenylene-
o-Phenanthroline Monohydrochloride Monohydrate, diamine Hydrochloride.
C12H8N2 HCl H2O234.69 [3829-86-5]Use a suitable p-Phenylenediamine Hydrochloride (1,4-Diaminobenzene
grade. Dihydrochloride), C6H8N2 2HCl181.06White to pale tan
Phenol [108-95-2]Use ACS reagent grade. crystals or crystalline powder, turning red on exposure to
Phenol Red, Sodium, C19H13O5SNa376.4 air. Freely soluble in water; slightly soluble in alcohol and in
[34487-61-1]Red to brown powder. Use ACS reagent ether. Preserve in well-closed containers, protected from
grade. light.
PhenolsulfonphthaleinUse Phenol Red (see Indicators
Insoluble matterDissolve 1 g in 10 mL of water: the so-
under Indicators and Indicator Test Papers).
lution is clear and complete.
Phenoxybenzamine Hydrochloride [N-(2-Chloroethyl)-N-
Molar absorptivity (see Spectrophotometry and Light-
(1-methyl-2-phenoxyethyl)benzylamine Hydrochloride],
scattering 851)Dissolve 60 mg in 100.0 mL of water, and
C18H22ClNO HCl340.29 [63-92-3]White, crystalline
mix. Pipet 2 mL of this solution into a 50-mL volumetric
powder.
flask, dilute with pH 7 buffer solution to volume, and mix.
The molar absorptivity of this solution, at 239 nm, is not
AbsorptivityIts absorptivity, 1%, 1 cm, in the range of less than 9000.
272 nm to 290 nm, in chloroform solution is about 178. Phenylglycine (D()-2-Phenylglycine),
3-Phenoxybenzoic Acid, C13H10O3214.22 (C6H5CH(NH2)COOH)151.17 [875-74-1]Use a suita-
[3739-38-6]Use a suitable grade. ble grade.
Phenylhydrazine, C6H5NHNH2108.14 [100-63-0]
A colorless, or slightly yellowish, highly refractive liquid.
2-Phenoxyethanol, C6H5OCH2CH2OH138.16
[NOTEProtect from light, and distill under reduced pres-
[122-99-6]Colorless, slightly viscous liquid. Soluble in
sure shortly prior to use.]
water. Miscible with alcohol, with acetone, and with glyc-
erin. Density: about 1.107. Congealing temperature 651: not below 16.
AssayTo 2 g, accurately weighed, add 10 mL of a Insoluble matterShake 1 mL with 20 mL of diluted ace-
freshly prepared solution made by dissolving 25 g of acetic tic acid: the resulting solution is clear or practically so.
anhydride in 100 g of anhydrous pyridine. Swirl to mix the
Residue on ignition (Reagent test)Ignite 1 mL with
liquids, heat on a steam bath for 45 minutes, add 10 mL of
0.5 mL of sulfuric acid: the residue weighs not more than
water, heat for 2 additional minutes, and cool. Add 10 mL
1 mg (0.1%).
of normal butyl alcohol, shake vigorously, add phenol-
Phenylhydrazine Hydrochloride, C6H5NHNH2 HCl
phthalein TS, and titrate with 1 N sodium hydroxide VS.
144.60 [59-88-1]White or yellowish crystals or powder.
Perform a blank test using the same quantities of the same
Soluble in water and in alcohol. Store in tight containers,
reagents, and in the same manner, and make any necessary
protected from light. Use a suitable grade with a content of
correction. Each mL of 1 N sodium hydroxide is equivalent
not less than 99%.
to 138.2 mg of C8H10O2. Not less than 99% is found.
Phenylmethylsulfonyl Fluoride, C7H7FO2S174.2
PhenolAdd 0.2 mL of it to 20 mL of water, mix, and to [329-98-6]White to faint yellow powder. Use a suitable
5 mL of the mixture add 0.2 mL of Millons reagent. Warm grade.
the solution at 60 for 90 seconds, and allow to stand: no [NOTEA suitable grade is available from Sigma-Aldrich,
pink or red color is produced within 1 minute. www.sigma-aldrich.com.]
Phenyl EtherSee Diphenyl Ether. 3-Phenylphenol (m-Phenylphenol), C6H5C6H4OH170.21
Phenyl Isocyanate, C6H5NCO119.12 [103-71-9] [580-51-8]White to off-white, crystalline powder.
Clear, colorless to straw-yellow liquid of medium volatility.
[CAUTIONPhenyl Isocyanate is a violent lacrimator, and the
vapor is highly toxic. Handle with care.]
AssayTransfer 250 mg, accurately weighed, to a glass- gas. The following conditions have been found suitable: a
stoppered, 250-mL flask. Exercise care to avoid loss by vola- 0.25-mm 30-m capillary column coated with G1; the in-
tilization, and avoid breathing the vapor. Add 20 mL of bu- jection port temperature is maintained at 250; the column
tylamine solution (25 g of butylamine diluted to 1000 mL temperature is maintained at 150 and programmed to rise
with dioxane previously dried over potassium hydroxide pel- 15 per minute to 250; and the detector temperature is
lets), insert the stopper in the flask, and allow to stand for maintained at 310. The area of the 3-phenylphenol peak is
15 minutes. Add a few drops of methyl red TS and 25 mL of not less than 98% of the total peak area.
water, and titrate the excess amine with 0.1 N sulfuric acid
VS. Perform a blank titration on 20 mL of the butylamine
Phloroglucinol, C6H3(OH)3 2H2O162.14
solution (see Residual Titrations 541). Subtract the volume
[6099-90-7]White or yellowish-white crystals or a crystal-
of 0.1 N sulfuric acid consumed in the test specimen titra-
line powder. Soluble in alcohol and in ether; slightly soluble
tion from that consumed in the blank titration. Each mL of
in water.
Insoluble in alcoholDissolve 1 g in 20 mL of alcohol: a NitrateDissolve 500 mg in 10 mL of water, and add
clear and complete solution results. about 10 mg of sodium chloride, 0.1 mL of indigo carmine
TS, and 10 mL of sulfuric acid: the blue color does not dis-
Melting range, Class Ia 741: between 215 and 219.
appear within 1 minute (about 0.01%).
Residue on ignition (Reagent test)Ignite 1 g with 0.5 mL
Sulfate (Reagent test, Method I )A 500-mg portion
of sulfuric acid: the residue weighs not more than 1 mg
shows not more than 0.1 mg of SO4 (0.02%).
(0.1%).
o-Phthalaldehyde (Phthalic Dicarboxaldehyde),
DiresorcinolHeat to boiling a solution of 100 mg in
C6H4(CHO)2134.13 [643-79-8]Use a suitable grade.
10 mL of acetic anhydride, cool the solution, and superim-
Phthalazine, C8H6N2130.15 [253-52-1]Yellow to
pose it upon 10 mL of sulfuric acid: no violet color appears
tan crystals.
at the zone of contact of the liquids.
Phloxine B (Acid red 92; Eosin 10B; 2,4,5,7-tetrabromo-
Phthalic Acid, C8H6O4166.13 [88-99-3]Use ACS
4,5,6,7-tetrachlorofluorescein disodium salt)
reagent grade.
C20H2Br4Cl4Na2O5829.63 [18472-87-2]Use a suitable
Phthalic Anhydride, C8H4O3148.12 [85-44-9]Use
grade with a dye content of not less than 80%, certified by
ACS reagent grade.
the Biological Stain Commission.
Phthalimide, C8H5NO2147.13 [85-41-6]White
Phosphatase Enzyme, AlkalineUse a suitable grade
powder.
from intestinal origin.
[NOTEA suitable grade is available from Worthington Assay
Biochemical Corp., www.worthington-biochem.com.]
MOBILE PHASEPrepare a mixture of isooctane and methyl-
Phosphatic EnzymeAn enzyme preparation of microbial
tert-butyl ether (88:12).
origin, high in both phosphatase and amylase activity, the
former being the property that renders it suitable for use in
the liberation of thiamine from its orthophosphate and py-
rophosphate esters. Light cream-colored or slightly gray
tains packing L3. The flow rate is about 2 mL per minute.
powder. Freely soluble in water. It hydrolyzes 300 times its
The area of the C8H5NO2 peak is not less than 99% of the
weight of starch in 30 minutes.
total peak area.
Amylase activityPlace in a test tube 5 mL of a 1 in 50
Melting range 741: between 233 and 235, with de-
solution of soluble starch in 0.2 M, pH 5 sodium acetate
composition.
buffer (containing 1.6 g of anhydrous sodium acetate in
2-Picoline, C6H7N93.13 [109-06-8]Colorless to
each L and sufficient glacial acetic acid to adjust to a pH of
yellowish liquid.
5), and add 4 mL of water. Mix, and place in a water bath
at 40. Add 1 mL of a solution containing 0.3 mg of the
phosphatic enzyme, mix, and note the exact time. After
30 minutes remove 1.0 mL of the mixture, and add it to
5.0 mL of 0.0005 N iodine in a 20- 150-mm test tube: a
2-mm 2-m glass column packed with 20% liquid phase
clear, red color results.
G16 on 80- to 100-mesh support S1C; the injection port
Phosphomolybdic Acid, approximately 20MoO3 P2O5
temperature is maintained at 140; the detector tempera-
51H2O3939.49 [11104-88-4]Use ACS reagent grade.
ture is maintained at 300; the column temperature is main-
Phosphoric Acid, H3PO498.00 [7664-38-2]Use
ACS reagent grade.
140. The area of the C6H7N peak is not less than 98% of
Phosphorous Acid (Phosphonic Acid), H3O3P82.00
[13598-36-2]Use a suitable grade with a content of not
Refractive index 831: 1.500 0.002 at 20. less than 99%.
Picric Acid (2,4,6-Trinitrophenol; Trinitrophenol), Phosphorus, Red, PAt. Wt. 30.97376A dark red
C6H2(OH)(NO2)3-1,2,4,6229.10 [88-89-1]Use ACS re- powder. Insoluble in water and in dilute acids; soluble in
agent grade. dehydrated alcohol.
Picrolonic Acid (3-Methyl-4-nitro-1-(p-nitrophenyl)-
Yellow phosphorusShake 20 g with 75 mL of carbon di-
5-pyrazolone), C10H8N4O5264.19 [550-74-3]Yellow to
sulfide in a glass-stoppered vessel, and allow to stand in the
brownish-yellow, crystalline powder. Slightly soluble in
dark overnight. Filter, and wash the residue with carbon di-
water; soluble in alcohol, in chloroform, in ether, in ben-
sulfide until the filtrate, collected in a graduated cylinder,
zene, and in solutions of alkali hydroxides.
measures 100 mL. Evaporate the solvent to 10 mL by im-
Melting range 741: between 115 and 117. mersing the cylinder in hot water. Dip a strip of cupric sul-
fate test paper in the remaining solvent: no more color is Residue on ignition (Reagent test): negligible, from
produced than in a similar strip dipped into 10 mL of solu- 200 mg.
tion in carbon disulfide containing 3 mg of yellow phos-
SensitivenessDissolve 25 mg in 10 mL of warm water
phorus (0.015% as P).
containing 0.1 mL of glacial acetic acid, and filter the solu-
Soluble substancesDigest 2 g with 30 mL of acetic acid tion, if necessary. Dissolve 100 mg of calcium chloride in
on a steam bath for 15 minutes. Cool, dilute with water to 250 mL of water, and mix. Heat 1 mL of the calcium chlo-
40 mL, and filter. Evaporate 20 mL of the filtrate on a steam ride solution in a test tube to about 60, then add to it
bath, and dry at 105 for 2 hours: the residue weighs not 1 mL of the picrolonic acid solution: a bulky precipitate
more than 6 mg (0.6%). forms in 5 minutes or less.
Phosphorus Pentoxide (Phosphoric Anhydride), P2O5 Pipemidic Acid (8-Ethyl-3,8-dihydro-5-oxo-2-(1-piper-
141.94 [1314-56-3]Use ACS reagent grade. azinyl)pyrido[2,3-d]-pyrimidine-6-carboxylic acid),
Phosphotungstic Acid, approximately 24WO3 P2O5 C14H17N5O3303.3 [51940-44-4]Use a suitable grade.
51H2O6624.84White or yellowish-green crystals or a Piperazine (Diethylenediamine), C4H10N2 6H2O
crystalline powder. Soluble in water, in alcohol, and in 194.23Use a suitable grade.
ether. Piperidine, C5H11N85.15 [110-89-4]Colorless liq-
uid. Miscible with water and with alcohol. Specific gravity: Insoluble matter (Reagent test): not more than 1 mg,
about 0.860. from 5 g (0.02%).
0.3 mg of Cl (0.03%).
Congealing range 651: between 12 and 15. Residue on ignition: not more than 0.75%.
[NOTESuitable grades are available as catalog number U
232, from J.T. Baker Chemical Co., www.jtbaker.com.]
between 104 and 106.
Potassium Acetate, KC2H3O298.14 [127-08-2]Use
Refractive index: about 1.454.
ACS reagent grade.
Platinic Chloride (Chloroplatinic Acid), H2PtCl6 6H2O
Potassium AlumUse Potassium Alum [see Potassium
517.90 [18497-13-7]Use ACS reagent grade Chloropla-
Alum (USP monograph)].
tinic Acid.
Potassium Arsenate Monobasic, KH2AsO4180.03
Polydimethylsiloxane, viscosity 0.65 centistokes (Hexa-
[7784-41-0]Use a suitable grade with a content of NLT
methyldisiloxane), (CH3)3SiOSi(CH3)3162.38
98%.
[107-46-0]Liquid. Freezes at about 0.
Potassium Bicarbonate, KHCO3100.12
Refractive index 831: about 1.3770.
Specific gravity 841: about 0.760. Potassium BiphosphateSee Potassium Phosphate, Mono-
Polyethylene Glycol 200, H(OCH2CH2)nOH in which the basic.
average value of n is 4average molecular weight 200 Potassium Biphthalate (Acid Potassium Phthalate; Phthalic
[25322-68-3]Clear, colorless or almost colorless, viscous, Acid Monopotassium Salt; Potassium Hydrogen Phthalate Acidi-
hygroscopic liquid. Very soluble in acetone and in alcohol; metric Standard), KHC6H4(COO)2204.22 [877-24-7]
practically insoluble in ether and in fatty oils. Use a suitable Use ACS reagent grade Potassium Hydrogen Phthalate, Aci-
grade. dimetric Standard.
Potassium Bisulfate, KHSO4136.17 [7646-93-7] Refractive index 831: 1.4590 at 20.
Fused, white, deliquescent masses or granules. Very soluble
Density: 1.127 at 25.
in water. When ignited, it evolves SO3 and H2O, changing
Viscosity: 4.3 centistokes at 98.9.
first to potassium pyrosulfate, then to sulfate.
Polyethylene Glycol 600 [25322-68-3]A clear, practi-
AcidityDissolve 4 g, accurately weighed, in 50 mL of
cally colorless, viscous liquid condensation polymer repre-
water, add phenolphthalein TS, and titrate with 1 N alkali: it
sented by H(OCH2CH2)nOH, in which n varies from 12 to
contains between 34% and 36%, calculated as H2SO4.
14. Its average molecular weight is about 600.
Insoluble matter and ammonium hydroxide precipitate It meets the requirements of all of the tests under Polyeth-
Dissolve 10 g in 100 mL of water, add methyl red TS, render ylene Glycol (NF monograph) except Limit of ethylene glycol
slightly alkaline with ammonia TS, boil for 1 minute, and and diethylene glycol.
digest on a steam bath for 1 hour. Pass through a tared Polyethylene Glycol 20,000 [25322-68-3]Molecular
filtering crucible, wash thoroughly, and dry at 105 for weight range: 15,00020,000. Hard, white, waxy solid, usu-
2 hours: the precipitate weighs not more than 1 mg ally supplied in flake form. Soluble in water with subsequent
(0.01%). gel formation.
For the following tests, prepare a Test solution as follows.
Viscosity of 25% solution 911Add 50.0 g of test speci-
Dissolve 6 g in 45 mL of water, add 2 mL of hydrochloric
men to a 250-mL wide-mouth, screw-cap jar containing
acid, boil gently for 10 minutes, cool, and dilute with water
150.0 g of water. Attach the cap securely to the jar, and roll
to 60 mL.
on a mechanical roller until the test specimen is completely
Heavy metals (Reagent test)To 30 mL of Test solution dissolved, in 2 to 4 hours. Allow the solution to stand until
add phenolphthalein TS, and neutralize with ammonia TS. all air bubbles have disappeared. Another 2 to 4 hours may
Add 0.5 mL of glacial acetic acid, dilute with water to be required. Adjust the temperature of the solution to 37.8
40 mL, and add 10 mL of hydrogen sulfide TS: any brown 0.1, and determine the kinematic viscosity on a suitable
color produced is not darker than that of a control contain- viscometer of the Ubbelohde type. The viscosity is not less
ing 10 mL of Test solution and 0.02 mg of added Pb than 100 centistokes.
(0.001%).
pH 791: between 6.5 and 8.0 in a solution (1 in
Iron 241To 5 mL of Test solution add 2 mL of hydro- 20). [NOTEA five-fold dilution of the test solution prepared
chloric acid, and dilute with water to 47 mL: the solution for the Viscosity of 25% solution test may be used.]
shows not more than 0.01 mg of Fe (0.002%).
Residue on ignition 281: not more than 0.7%, the
Potassium Bromate, KBrO3167.00 [7758-01-2]
use of sulfuric acid being omitted.
Polyoxyethylene (23) Lauryl Ether (Brij-35)Use a suita-
Potassium Bromide, KBr119.00 [7758-02-3]Use
ble grade.
ACS reagent grade.
[NOTEA suitable grade is available commercially as Brij-
Potassium CarbonateSee Potassium Carbonate, Anhy-
35.]
drous.
Polyoxyethylene (20) Sorbitan Monolaurate
Potassium Carbonate, Anhydrous, K2CO3138.21
[9005-64-5]Viscous, light yellow to yellow-green liquid.
Potassium Chlorate, KClO3122.55 [3811-04-9]
Polysaccharide Molecular Weight Standards
Polymaltotriose polymers of different weight-average molec-
Potassium Chloride, KCl74.55 [7447-40-7]Use
ular weight, MW, values ranging from 5,000 to 400,000 Da.
ACS reagent grade.
[NOTEA suitable set is available from Shodex (www.
Potassium Chloroplatinate, K2PtCl6485.99Heavy,
shodex.com) as Kit P-82.]
yellow powder. Soluble in hydrochloric acid and in nitric
Polystyrene CationExchange ResinSee Ca-
acid.
tionExchange Resin, Polystyrene.
AssayAccurately weigh about 300 mg, transfer to a PolytefUse Poly(tetrafluoroethylene).
600-mL beaker, add 20 mL of hydrochloric acid, and heat Polyvinyl Alcohol, (C2H4O)n [9002-89-5]White pow-
gently if necessary to achieve complete solution. Add zinc der. Soluble in water; insoluble in organic solvents.
granules, slowly, until no more dissolves, then add 2 mL of
pH 791: between 5.0 and 8.0, in a solution (1 in
hydrochloric acid, and digest for 1 hour on a steam bath to
25).
coagulate the reduced platinum. Add more acid, if neces-
Loss on dryingDry it at 110 to constant weight: it loses
sary, to ensure that all of the zinc has dissolved. Filter
not more than 5% of its weight.
through paper, rinsing the beaker with diluted hydrochloric
acid until all of the precipitate is transferred to the filter,
then wash with several small portions of water. Ignite the
filter in a tared crucible at 800 25 to constant weight.
Each mg of residue is equivalent to 1.0 mg of platinum. Not Potassium Metabisulfite (Potassium Disulfite; Potassium
less than 40% is found. Pyrosulfite), K2S2O5222.32 [16731-55-8]Use a suitable
Potassium Chromate, K2CrO4194.19 [7789-00-6] grade with a content of not less than 98%.
Use ACS reagent grade. Potassium Nitrate, KNO3101.10 [7757-79-1]Use
Potassium Cyanide, KCN65.12 [151-50-8]Use ACS reagent grade.
ACS reagent grade. Potassium Nitrite, KNO285.10 [7758-09-0]Use
Potassium Dichromate, K2Cr2O7294.18 ACS reagent grade.
[7778-50-9]Use ACS reagent grade. Potassium Perchlorate, KClO4138.55
[NOTEPotassium dichromate of a quality suitable as a [7778-74-7]Use ACS reagent grade.
primary standard is available from the National Institute of Potassium Periodate (Potassium meta-Periodate), KIO4
Standards and Technology, Washington, DC, www.nist.gov, 230.00 [7790-21-8]Use ACS reagent grade.
as standard sample No. 136.] Potassium Permanganate, KMnO4158.03
Potassium Ferricyanide, K3Fe(CN)6329.24 [7722-64-7]Use ACS reagent grade.
[13746-66-2]Use ACS reagent grade. Potassium Persulfate, K2S2O8270.32 [7727-21-1]
Potassium Ferrocyanide, K4Fe(CN)6 3H2O422.39 Use ACS reagent grade Potassium Peroxydisulfate.
[14459-95-1]Use ACS reagent grade. Potassium Phosphate, Dibasic (Dipotassium Hydrogen
Potassium HyaluronateWhite to cream-colored pow- Phosphate; Dipotassium Phosphate), K2HPO4174.18
der. Freely soluble in water. Store in a tight container, in a [7758-11-4]Use ACS reagent grade.
refrigerator. Potassium Phosphate, Dibasic, Trihydrate (Dipotassium
Hydrogen Phosphate Trihydrate; Dipotassium Phosphate),
Inhibitor contentPrepare as directed in the Assay under
K2HPO4 3H2O228.22 [16788-57-1]Use a suitable
Hyaluronidase for Injection (USP monograph) a quantity of
grade with a content of NLT 99.0%.
Standard solution containing 1 USP Hyaluronidase Unit in
Potassium Phosphate, Monobasic (Potassium
each mL, and a similar quantity of acetate-buffered Standard
Biphosphate; Potassium Dihydrogen Phosphate), KH2PO4
solution using as the solvent 0.1 M, pH 6 sodium acetate
buffer (prepared by diluting the 0.2 M buffer prepared as
[NOTECertified Potassium Dihydrogen Phosphate is avail-
directed below with an equal volume of water). Prepare
able from the National Institute of Standards and Technol-
from the potassium hyaluronate under test 10 mL of Potas-
ogy, Washington, DC, www.nist.gov, as standard sample
sium hyaluronate stock solution, and dilute 2 mL of it with
No. 186.]
the specified Phosphate buffer solution to make a Hyaluronate
Potassium Phosphate, Tribasic, K3PO4212.27
solution. In the same way, and concurrently, dilute a second
[7778-53-2]Deliquescent, orthorhombic crystals. Use ACS
2-mL portion of the stock solution with 0.2 M, pH 6 sodium
reagent grade.
acetate buffer (containing 16.4 g of anhydrous sodium ace-
Potassium Pyroantimonate (Potassium hexahydroxyan-
tate and 0.45 mL of glacial acetic acid in each 1000 mL).
timonate), KSb(OH)6262.90 [12208-13-8]White crys-
Place 0.50-mL portions of the Hyaluronate solution in each
tals or a white, crystalline powder. Sparingly soluble in
of four 16- 100-mm test tubes, and place 0.50-mL por-
water. Use a suitable grade.
tions of the acetate-buffered Hyaluronate solution in two sim-
Potassium Pyrophosphate, K4P2O7330.34
ilar tubes. To two of the four tubes containing Hyaluronate
[7320-34-5]Colorless, deliquescent granules. Freely soluble
solution add 0.50 mL of Diluent for hyaluronidase solutions,
in water; insoluble in alcohol.
prepared as directed in the Assay under Hyaluronidase for
Potassium Pyrosulfate [7790-62-7]Usually available as
Injection (USP monograph). To the remaining two tubes, on
a mixture of potassium pyrosulfate (K2S2O7) and potassium
a rigid schedule, at 30-second intervals, add 0.50 mL of
bisulfate (KHSO4). Use ACS reagent grade.
Standard solution. Similarly, to the two tubes containing ace-
Potassium Sodium Tartrate, KNaC4H4O6 4H2O282.22
tate-buffered Hyaluronate solution add at 30-second intervals
0.50-mL portions of acetate-buffered Standard solution. Then
Potassium Sulfate, K2SO4174.26 [7778-80-5]Use
proceed as directed in the second paragraph for Procedure,
ACS reagent grade.
beginning with Mix the contents, as far as Plot the aver-
Potassium Tellurite (Potassium Tellurate IV), K2TeO3
age. The reduction in absorbance of acetate-buffered Hy-
253.79 [7790-58-1]White, granular powder. Soluble in
aluronate solution is not less than 25% of that observed in
water. Its solution is alkaline.
the Hyaluronate solution.
AssayWeigh accurately about 120 mg, transfer to a
Turbidity productionThe average absorbance of the solu-
beaker, and dissolve in a mixture of 10 mL of nitric acid,
tions in the two tubes containing Hyaluronate solution and
10 mL of sulfuric acid, and 25 mL of water. Heat to boiling,
Diluent for hyaluronidase solutions prepared in the test for
and boil until copious fumes of sulfur trioxide are evolved.
Inhibitor content is not less than 0.26 at a wavelength of
Cool, cautiously add 100 mL of water, heat to boiling, add
640 nm in a suitable spectrophotometer using a 1-cm cell.
6 g of sodium fluoride, and titrate the hot solution with
Potassium Hydrogen Sulfate, KHSO4136.17White
0.1 N potassium permanganate VS. Each mL of 0.1 N potas-
crystals. Soluble in water.
sium permanganate is equivalent to 12.69 mg of K2TeO3.
Not less than 98% is found.
Potassium Hydroxide, KOH56.11 [1310-58-3]Use
ACS reagent grade.
0.1 mg of Cl (0.01%).
Potassium Iodate, KIO3214.00 [7758-05-6]Use
Potassium Thiocyanate, KSCN97.18 [333-20-0]
ACS reagent grade.
Potassium Iodide, KI166.00 [7681-11-0]Use ACS
Potato StarchSee Starch, Potato.
reagent grade.
Propionaldehyde, C3H6O58.08 [123-38-6]Use a
suitable grade.
Propionic Anhydride, C6H10O3130.14 [123-62-6]
Colorless liquid. Is decomposed by water. Soluble in metha-
nol, in alcohol, in ether, and in chloroform.
AssayAccurately weigh about 350 mg into a tared,
glass-stoppered flask containing 50 mL of dimethylform-
amide previously neutralized to the thymol blue endpoint
with 0.1 N sodium methoxide in methanol VS. Titrate with
0.1 N sodium methoxide in methanol VS to the thymol blue
endpoint. Perform a blank determination, and make any
necessary correction. Each mL of 0.1 N sodium methoxide is Glucose standard solutionDry anhydrous glucose crystals
equivalent to 13.014 mg of C6H10O3. Not less than 97.0% is under less than 50-mm Hg at 60 for 5 hours, and calculate
found. the water content. Transfer 10.00 g of dried glucose to a 1-L
volumetric flask, dissolve in and dilute with water to vol-
ume, and mix. Transfer 10.0 mL to a 1-L volumetric flask
20.
and dilute to volume with water. Each mL contains 100 g
Propiophenone, C9H10O134.18 [93-55-0]Use a
of glucose.
suitable grade.
iso-Propyl AlcoholSee Isopropyl Alcohol. Pullulanase diluentDilute pullulanase with Buffer solution
n-Propyl Alcohol (1-Propanol), CH3CH2CH2OH60.10 B, pH 6.0 to prepare a solution having the enzyme activity
[71-23-8]Use ACS reagent grade. of about 0.2 units per mL. [NOTEThe measurement range
Propylamine Hydrochloride (1-Propanamine hydrochlo- is between 0.1 and 0.4 units per mL.] Record the dilution
ride; n-Propylamine hydrochloride), C3H9N HCl95.57 factor (FPD). This diluent is used as a diluted enzyme solu-
[556-53-6]Use a suitable grade with a content of not less tion.
than 99%.
ProcedureTransfer 4 mL of Substrate to a test tube and
Protein Molecular Weight StandardAlso known as
add 0.5 mL of Buffer solution A, pH 5.0,mix, and incubate at
protein molecular weight markers (for SDS-PAGE) and con-
30. Add 0.5 mL of Pullulanase diluent, and mix thoroughly.
sists of a mixture of several proteins of well-defined molecu-
After 30 seconds, transfer 1 mL of this solution to a test tube
lar weights. The products are generally available in a suita-
labeled as Pullulan test solution 1, and add 2 mL of Somo-
ble buffer containing a suitable reducing agent (generally,
gyi reagent, and mix. After 30 minutes and 30 seconds,
100 mM DTT), a preservative (for example, sodium azide),
transfer 1 mL of the mixture of Substrate and Pullulanase di-
and 50% glycerol to prevent freezing. Use a suitable grade.
luent to a second test tube labeled as Pullulan test solution
Store at 20.
2, add 2 mL of Somogyi reagent, and mix. In a third test
Protein Standard Solution (8 g/dL)A solution contain-
tube labeled as Standard blank, mix 2 mL of Somogyi rea-
ing 5 g of Albumin Human and 3 g of human gamma glob-
gent and 1 mL of water. In a fourth test tube labeled as
ulin per dL.
Glucose standard solution, mix 2 mL of Somogyi reagent
[NOTEA suitable grade is available as Protein Standard
and 1 mL of Glucose standard solution, and add 1 mL of
Solution, catalog number 540-10, from Sigma-Aldrich,
water. Incubate the fourth test tube in a boiling water bath
for exactly 10 minutes. Remove the tube and allow it to
Protocatechuic Acid (3,4-Dihydroxybenzoic acid),
cool in cold running water. Add 2 mL of Nelson reagent, mix
well, and allow the solution to stand for at least 15 minutes.
Pullulanase (Amylopectin-6-gluconohydrolase)
Add 5 mL of water to each of the four test tubes, and mix
[9075-68-7]An enzyme obtained from Kleibsiella
thoroughly. Determine the absorbance at 520 nm of the
pneumoniae. It contains not less than 30 units per mg of
Standard blank, Ablank, of the Glucose standard solution, AStd,
protein. One unit defined for enzymatic activity will liberate
of the Pullulan test solution 1, A0, and of the Pullulan test
1.0 mol of maltotriose (measured as glucose) from pullulan
solution 2, A30, using water as the blank. One unit is defined
per minute at pH 5.0 at 30. It can be suspended in 3.2 M
as the enzymatic activity that produces 1 mol of maltotri-
ammonium sulfate solution, pH 6.2.
ose (measured as glucose) from pullulan per minute. Calcu-
Measurement of relative pullulanase activity late the pullulanase activity, PA, in units per mL, using the
formula:
DETERMINATION OF PULLULANASE ACTIVITY
SubstrateDissolve pullulan
1
in water to make a 1.25%
PA = [(A30 A0)/(AStd Ablank)] 0.185 FPD
(w/v) solution. [NOTEAdd pullulan to the water. Clumping
will occur if water is added to pullulan.]
MEASUREMENT OF PROTEIN AMOUNT (MEASURED AS ALBUMINOID
Buffer solution A, pH 5.0Add 0.1 M disodium phosphate
AMOUNT) FOR THE CALCULATION OF SPECIFIC ACTIVITY
(27 g of dibasic sodium phosphate in each L of the solution)
Reagent APrepare a solution having known concentra-
to 0.1 M citric acid (21 g of citric acid in each L of the
tions of about 0.1 N sodium hydroxide and about 0.2 M
solution) to adjust pH to 5.0.
sodium carbonate.
Buffer solution B, pH 6.0Add diluted acetic acid to 1 M
Reagent BTransfer 0.5 g of cupric sulfate and 1.0 g of
sodium acetate (136 g of sodium acetate in each L of solu-
sodium citrate dihydrate to a 100-mL volumetric flask, dis-
tion) to adjust the pH to 6.0. Dilute with water to prepare
solve in and dilute with water to volume, and mix.
the final buffer solution as 0.01 M acetic acid buffer, pH
Lowry solutionMix Reagent A and Reagent B at the pro- 6.0.
portion of 50:1.
Somogyi reagentAdd 54 g of disodium phosphate
Diluted Folin-Ciocalteus phenol reagent (for albuminoid heptahydrate or 28 g of anhydrous disodium hydrogen
quantification)Prepare a two-fold dilution of 2 N Folin-Cio- phosphate and 40 g of potassium sodium tartrate to about
calteus phenol reagent commercially available or prepare a 650 mL of water or about 700 mL for anhydrous disodium
solution by making an appropriate dilution from Folin-Cio- hydrogen phosphate. Add 100 mL of 1 N sodium hydroxide
calteu Phenol TS (see Method 2 in Biotechnology-Derived Arti- to this solution and mix. Add 80 mL of 10% cupric sulfate
clesTotal Protein Assay 1057). to the solution, and mix. Heat until any solid is completely
dissolved. Add 180 g of anhydrous sodium sulfate to the Bovine serum albumin standard stock solutionTransfer
solution and adjust the volume to 1 L. Allow the solution to 0.05 g of bovine serum albumin to a 500-mL volumetric
stand at room temperature for 1 or 2 days to let insoluble flask, dissolve in and dilute with water to volume, and mix.
matter precipitate. Filter the solution with standard filter pa- It contains 100 g of bovine serum albumin per mL.
per, and keep the solution in a brown bottle with a ground-
Standard solutionsUsing appropriate dilutions of Bovine
glass stopper.
serum albumin standard stock solution in water, prepare five
Nelson reagentDissolve 50 g of ammonium molybdate Standard solutions having concentration equally spaced be-
in 900 mL of water. Add 42 g of sulfuric acid, and mix. Dis- tween 5 and 100 g of bovine serum albumin per mL.
solve 6 g of sodium arsenate or 3.6 g of monobasic potas-
Test solutionDilute pullulanase with Buffer solution B, pH
sium arsenate in 50 g of water. Allow the solution to stand
6.0 in order to obtain a solution having a concentration
in a brown bottle with a ground-glass stopper at 37 for 1
between 60 and 70 g of albuminoid per mL. [NOTEWater
or 2 days.
can be used as diluent.] Record the dilution factor, FTS.
1
A suitable supplier for pullulan is www.hayashibara-intl.com.
Blank solutionUse water. Melting range 741: between 149 and 153 over a
2 range.
ProcedureTo 0.3 mL in separate tubes of the Standard
Pyridine, C5H5N79.10 [110-86-1]Use ACS reagent
solutions, the Test solution, and the Blank solution, add 3 mL
grade.
of Lowry solution, and mix. Allow to incubate at room tem-
Pyridine, Dried [110-86-1]Use ACS reagent grade.
perature for 10 minutes. Add 0.3 mL of Diluted Folin-Cio-
Pyridoxal Hydrochloride, C8H9NO3 HCl203.62
calteus phenol reagent to each tube, mix immediately, and
[65-22-5]White to slightly yellow crystals or crystalline
allow to stand at room temperature for 60 minutes. Deter-
powder. Gradually darkens on exposure to air or sunlight.
mine the absorbances of the Standard solutions and the Test
One g dissolves in about 2 mL of water and in about 25 mL
solution at the wavelength of maximum absorbance at
of alcohol. Insoluble in acetone, in chloroform, and in ether.
about 750 nm, using the Blank solution as the blank.
Its solutions are acid (pH about 3).
Calculation[NOTEThe relationship of absorbance to
Melting range 741: between 171 and 175 with
protein concentration is nonlinear; however, if the standard
some decomposition.
curve concentration range is sufficiently small, it will ap-
proach linearity.] Using linear regression method, plot the Residue on ignition (Reagent test): not more than 0.1%.
absorbances of the Standard solutions versus the protein (bo-
vine serum albumin) concentrations, in g per mL, and de-
termine the best fit curve. Using the plot, determine the
concentration, Calbuminoid, in g per mL, of protein (albumi-
noid amount) in the Test solution. Calculate the albuminoid
105 for 2 hours: between 6.7% and 7.1% of N is found.
concentration, in mg per mL, in the pullulanase taken by
Chloride contentAccurately weigh about 500 mg, previ-
the formula:
ously dried at 105 for 2 hours, and dissolve in 50 mL of
water. Add 3 mL of nitric acid and 50.0 mL of 0.1 N silver
Cprotein = (Calbuminoid FTS)/1000
nitrate VS, then add 5 mL of nitrobenzene, shake for about
2 minutes, add ferric ammonium sulfate TS, and titrate the
Calculate the specific activity, SA, in units per mg, of pullu-
excess silver nitrate with 0.1 N ammonium thiocyanate VS:
lanase using the formula:
each mL of 0.1 N silver nitrate is equivalent to 3.545 mg of
Cl. Between 17.2% and 17.7% is found. SA = PA/Cprotein
Pyridoxal 5-Phosphate, 4-CHOC5HN-2-CH3, 3-OH,
5-CH2PO4H2 H2O265.16 [41468-25-1]Light yellow PumiceA substance of volcanic origin consisting chiefly
powder. Use a suitable grade. of complex silicates of aluminum and alkali metals. Occurs
Pyridoxamine Dihydrochloride, C8H12N2O2 2HCl as very light, hard, rough, porous, gray masses, or as a
241.11 [524-36-7]White to slightly yellow crystals or gray-colored powder. Is insoluble in water and is not at-
crystalline powder. Gradually darkens on exposure to air or tacked by diluted acids.
sunlight. One g dissolves in about 1 mL of water and in
Acid- and water-soluble substancesBoil 2.0 g of pow-
about 60 mL of alcohol. Insoluble in chloroform and in
dered pumice with 50 mL of diluted hydrochloric acid under
ether. Its solutions are acid.
a reflux condenser for 30 minutes. Cool, and filter. To half of
Melting range 741: between 225 and 230, with the filtrate add 5 drops of sulfuric acid, evaporate to dry-
some decomposition. ness, ignite, and weigh: the residue weighs not more than
60 mg (6.0%). Residue on ignition (Reagent test): not more than
Purine, C5H4N4120.11 [120-73-0]White to off- 0.15%.
white powder.
Melting range 741: between 214 and 217. not more than 0.5% of its weight.
A single spot is exhibited when it is examined by thin-
layer chromatography, with the use of plates coated with
105 for 2 hours: between 11.3% and 11.8% of N is found.
consisting of butyl alcohol, water, and glacial acetic acid
Chloride contentDetermine as directed in the test for
(60:25:15).
Chloride content under Pyridoxal Hydrochloride: between
Putrescine Dihydrochloride, C4H12N2 2HCl161.07
29.1% and 29.6% of Cl is found.
[333-93-7]White, crystalline powder. Use a suitable grade.
1-(2-Pyridylazo)-2-naphthol, C15H11N3O249.27
Pyrazole, C3H4N2 [288-13-1]White to pale yellow crys-
[85-85-8]Stable, orange-red crystals. Soluble in alcohol
tals or crystalline powder. Soluble in water, in alcohol, and
and in hot solutions of dilute alkalies; slightly soluble in
in ether.
water.
Pyrene, C16H10202.25 [129-00-0]White to light
SensitivenessAdd 0.1 mL of a 1 in 1000 solution of it in
yellow crystals.
alcohol to a mixture of 10 mL of water and 1 mL of a buffer
solution prepared by mixing 80 mL of 0.2 M acetic acid and
100-mL volumetric flask, dissolve in methanol, dilute with
20 mL of sodium acetate solution (8.2 in 100), and mix. To
methanol to volume, and mix. Transfer 2.0 mL of this solu-
this solution add 1 mL of a mixture of 1 mL of cupric sulfate
tion to a 100-mL volumetric flask, dilute with methanol to
TS and 2 mL of water, and mix: the color changes from
volume, and mix. Using a suitable spectrophotometer, 1-cm
yellow to red.
cells, and methanol as the blank, record the absorbance of
4-(2-Pyridylazo)resorcinol (PAR), C11H9N3O2, free acid;
the solution at the wavelength of maximum absorbance at
C11H8N3NaO2, monosodium salt215.21, free acid
about 238 nm. From the observed absorbance, calculate the
237.21, monosodium salt [1141-59-9,free acid;
absorptivity (see Spectrophotometry and Light-scattering
16593-81-0,monosodium salt]Use ACS reagent grade.
851): the absorptivity is not less than 432.9, correspond-
3-(2-Pyridyl)-5,6-di(2-furyl)-1,2,4-triazine-5,5-
ing to not less than 98% of C16H10.
disulfonic Acid, Disodium Salt (3-(2-Pyridyl)-5,6-bis(5-sulfo-
2-furyl)-1,2,4-triazine, Disodium Salt Hydrate),
C16H8N4Na2O8S2494.37 [79551-14-7]Use a suitable
grade.
[NOTEA suitable grade is available as product number dilute with water to 47 mL: the solution shows not more
P4272 from Sigma-Aldrich, 1-800-558-9160; www.sigma- than 0.01 mg of Fe (0.002%).
aldrich.com.] QuinoneSee p-Benzoquinone.
Pyrogallol, C6H3(OH)3126.11 [87-66-1]Use ACS RayonUse Purified Rayon (General Monograph).
reagent grade. Direct Red 80, C45H26N10Na6O21S61373.07
Pyrrole, C4H5N67.09 [109-97-7]Clear liquid, col- [2610-10-8]Red powder. Soluble in water; poorly soluble
orless when freshly distilled, becoming yellow in a few days. in alcohol. Use a suitable grade.
Specific gravity: about 0.94. Insoluble in water; soluble in Red-Cell Lysing AgentThe reagent is available as a so-
alcohol, in benzene, and in ether. lution containing 0.33% potassium cyanide and 0.11% so-
dium nitroprusside and a quarternary ammonium salt as the
surface active agent (5.5%).
between 128 and 132.
[NOTEThe reagent is manufactured by Coulter Electron-
Pyruvic Acid, CH3COCOOH88.06 [127-17-3]Col-
ics Diagnostics, Hialeah, FL and is available from many sup-
orless to light yellow liquid. Miscible with water, with alco-
pliers under the name of Zapoglobin
(or Zap-oglobin
).]
hol, and with ether.
Red PhosphorusSee Phosphorus, Red.
Resazurin (Sodium), C12H6NNaO4251.17
AssayAccurately weigh about 1 g, transfer to a suitable
[62758-13-8]A brownish-purple, crystalline powder. One
container, and add 100 mL of water. Mix, add phenol-
g dissolves in 100 mL of water, forming a deep-violet-
phthalein TS, and titrate with 0.5 N sodium hydroxide VS.
colored solution.
Each mL of 0.5 N sodium hydroxide is equivalent to
Hydrogen sulfide and other compounds containing the
44.03 mg of CH3COCOOH: not less than 98% of
thiol group decolorize solutions of resazurin sodium, form-
CH3COCOOH is found.
ing dihydroresorufin. When the decolorized solution is
Quantitative Filter PaperSee Filter Paper, Quantitative.
shaken in the presence of air, a rose color develops as a
Quinhydrone, C6H4(OH)2 C6H4O2218.21
result of the formation of resorufin.
[106-34-3]Green crystals having a metallic luster. Slightly
Retinyl Palmitate, C36H60O2524.9Yellow liquid.
soluble in cold water; soluble in hot water, in alcohol, and
Assay
in ether.
Mobile phasePrepare a mixture of acetonitrile and tetra-
hydrofuran (55:15).
glass-stoppered flask, add 50 mL of 1 N sulfuric acid and 3 g
ProcedureInject about 10 L into a suitable liquid chro-
of potassium iodide, insert the stopper in the flask, and
shake until dissolved. Titrate the liberated iodine with 0.1 N
sodium thiosulfate VS, adding 3 mL of starch TS as the
tains packing L1. The flow rate is about 1 mL per minute.
endpoint is approached. Each mL of 0.1 N sodium thiosul-
The area of the C36H60O2 peak is not less than 93% of the
fate is equivalent to 5.405 mg of quinone (C6H4O2). Be-
total peak area.
tween 49.0% and 51.0% is found.
Reverse TranscriptaseUse a suitable grade.
Alcohol-insoluble matterDissolve 10 g in 100 mL of hot
[NOTEA suitable grade is available from BD Biosciences,
alcohol, filter through a suitable tared crucible of fine poros-
www.bdbiosciences.com.]
ity, and wash with hot alcohol until the last washing is col-
Rhodamine B (Tetraethylrhodamine), C28H31ClN2O3
orless. Dry at 105, cool in a desiccator, and weigh: the
479.01 [81-88-9]Green crystals or a reddish-violet
residue weighs not more than 1.0 mg (0.010%).
powder. Very soluble in water, yielding a bluish-red solution
that is strongly fluorescent when dilute. Very soluble in alco-
0.050%, a 2.0-g test specimen being used. Save the resi-
hol; slightly soluble in dilute acids and in alkali solutions. In
due.
strong acid solution, it forms a pink complex with antimony
SulfateTransfer 1 g to a platinum crucible, add 10 mL that is soluble in isopropyl ether.
of hot water and 0.5 g of sodium carbonate, evaporate to
Clarity of solutionIts solution (1 in 200) is complete and
dryness, and ignite, protected from the sulfur in the flame,
clear.
until the residue is nearly white. Cool, add 20 mL of water
Residue on ignition (Reagent test)Ignite 1 g with 1 mL
and 1 mL of 30 percent hydrogen peroxide, boil gently for a
of sulfuric acid: the residue weighs not more than 2 mg
few minutes, add 2 mL of hydrochloric acid, and evaporate
(0.2%).
on a steam bath to dryness. Cool, dissolve the residue in
Rhodamine 6G, C28H31C1N2O3479.02 [989-38-8]
20 mL of water, filter, and to the filtrate add 1 mL of 1 N
hydrochloric acid and 3 mL of barium chloride TS: any tur-
Ribonuclease InhibitorUse a suitable grade.
bidity produced within 10 minutes does not exceed that in
[NOTEA suitable grade is available from BD Biosciences,
a control containing 0.2 mg of added SO4 and 0.5 mg of
www.bdbiosciences.com.]
sodium carbonate, 1 mL of 30 percent hydrogen peroxide,
Rose Bengal Sodium (Disodium Salt of 4,5,6,7-Tetra-
and 2 mL of hydrochloric acid previously evaporated on a
chloro-2,4,5,7-tetraiodofluorescein), C20H2Cl4I4Na2O5
steam bath to dryness (0.02%).
1017.64 [632-69-9]Fine, rose-colored crystals or crys-
Heavy metalsTo the residue retained from the test for
talline powder. Soluble in water.[NOTERender commercially
Residue on ignition add 2 mL of hydrochloric acid and
available material suitably pure by the following treatment.
0.5 mL of nitric acid, and evaporate on a steam bath to
Dissolve 8 g in 200 mL of water, and adjust to a pH be-
dryness. Dissolve the residue in 30 mL of hot water contain-
tween 10 and 11, using short-range pH indicator paper.
ing 1 mL of 1 N hydrochloric acid, cool, dilute with water to
Add 200 mL of acetone, while stirring gently, then add di-
40 mL, and mix. Dilute 20 mL of this solution (retain the
lute hydrochloric acid (1 in 10), while continuing to stir,
rest of the solution) with water to 25 mL, adjust to a pH
until the pH of the solution reaches 4.0. Add 400 mL more
between 3.0 and 4.0 by the addition of 1 N acetic acid or
of water, with stirring, and continue the stirring for 5 min-
6 N ammonium hydroxide as necessary, dilute with water to
utes. Filter the crystals on a filtering funnel, and return the
40 mL, and add 10 mL of freshly prepared hydrogen sulfide
crystals to the beaker used for crystallization. Recrystallize
TS: any brown color produced does not exceed that in a
three more times in the same manner, and dry the crystals
control containing 0.02 mg of added Pb (0.002%).
at 110 for 12 hours. Store in an amber bottle in a refrigera-
Iron 241To 10 mL of the solution retained from the tor at a temperature between 2 and 8. Prepare this rea-
test for Heavy metals add 2 mL of hydrochloric acid, and gent fresh monthly.]
Chromatographic purityDissolve 100 mg of rose bengal
sodium, prepared as described above, in 100 mL of water,
and apply 10 L of the solution on suitable chromato- [NOTEA suitable grade is available as Ottawa Standard
graphic paper. Develop the chromatogram by ascending Sand from Thomas Scientific, 99 High Hill Road at I-295,
chromatography, using a mixture of 1 part of dilute alcohol P.O. Box 99, Swedesboro, NJ 08085-0099.]
(1 in 4) and 1 part of dilute stronger ammonia water (1 in Sand, WashedIt may be prepared as follows. Digest
12). Examine the chromatogram in daylight and under UV clean, hard sand at room temperature with a mixture of
light (360 nm): no colored or fluorescent spot is visible 1 part of hydrochloric acid and 2 parts of water (about 13%
other than the rose bengal sodium spot. of HCl) for several days, or at an elevated temperature for
[NOTEA suitable grade is available commercially as Sil- several hours. Collect the sand on a filter, wash with water
ica Gel H.] until the washings are neutral and show only a slight reac-
Ruthenium Red (Ruthenium Oxychloride, Ammoniated), tion for chloride, and finally dry. Washed sand meets the
Ru2(OH)2Cl4 7NH3 3H2O551.23 [11103-72-3]A following tests.
brownish-red to dark purple powder. Soluble in water.
Substances soluble in hydrochloric acidDigest 10 g with a
S DesignationsSee supports for gas chromatography
mixture of 10 mL of hydrochloric acid and 40 mL of water
under Reagents, Chromatographic Columns.
on a steam bath for 4 hours, replacing from time to time
SaccharoseUse Sucrose (NF monograph).
the water lost by evaporation. Filter, and to 25 mL of the
Safranin O [477-73-6]Dark red powder consisting of a
filtrate add 5 drops of sulfuric acid, evaporate, and ignite to
mixture of 3,7-diamino-2,8-dimethyl-5-phenylphenazinium
constant weight: the residue weighs not more than 8 mg
chloride, C20H19ClN4350.85, and 3,7-diamino-2,8-di-
(0.16%).
methyl-5-o-tolylphenazinium chloride, C21H21ClN4364.88
Chloride (Reagent test)Shake 1 g with 20 mL of water
Sparingly soluble in 70 percent alcohol yielding a clear red
for 5 minutes, filter, and add to the filtrate 1 mL of nitric
solution with a yellowish-red fluorescence.
acid and 1 mL of silver nitrate TS: any turbidity produced
Identification
corresponds to not more than 0.03 mg of Cl (0.003%).
A: To 10 mL of a 0.5% w/v solution add 5 mL of hydro- Sawdust, PurifiedIt may be prepared as follows. Extract
chloric acid: a bluish violet solution is produced. sawdust in a percolator, first with sodium hydroxide solution
(1 in 100), and then with dilute hydrochloric acid (1 in 100)
B: To 10 mL of a 0.5% w/v solution add 5 mL of so-
until the acid percolate gives no test for alkaloid with mer-
dium hydroxide solution (1 in 5): a brownish-red precipitate
curic-potassium iodide TS or with iodine TS. Then wash with
is produced.
water until free from acid and soluble salts, and dry. Purified
C: To 100 mg add 5 mL of sulfuric acid: a green solu-
sawdust meets the following test.
tion is produced, which, on dilution, changes to blue and
AlkaloidsTo 5 g of purified sawdust contained in a flask
finally to red.
add 50 mL of a mixture of 2 volumes of ether and 1 volume
Absorption characteristicsDissolve 50 mg in 250 mL of
of chloroform and 10 mL of ammonia TS, and shake fre-
50 percent alcohol. Dilute 3 mL of this solution with 50 per-
quently for 2 hours. Decant 20 mL of the clear, ether-chloro-
cent alcohol to 200 mL. Determine the absorbance, in a
form liquid, and evaporate to dryness. Dissolve the residue
1-cm cell, with a suitable spectrophotometer. The absorb-
in 2 mL of dilute hydrochloric acid (1 in 12), and divide into
ance maximum is in the range of 530 to 533 nm; the ratio
two portions. To 1 portion add mercuric-potassium iodide
(P 15)/(P + 15) is between 1.10 and 1.32, in which P is
TS, and to the other add iodine TS: no turbidity is produced
the wavelength of maximum absorbance.
in either portion.
Scandium Oxide, Sc2O3137.91 [12060-01-1]Fine,
10,214-8 from Sigma-Aldrich, www.sigma-aldrich.com.]
white powder.
Salicylaldazine, C14H12N2O2240.26Use a suitable
Secondary Butyl AlcoholSee Butyl Alcohol, Secondary.
grade or prepare as follows. Dissolve 300 mg of hydrazine
Selenious Acid (Selenous Acid), H2SeO3128.97
sulfate in 5 mL of water, add 1 mL of glacial acetic acid and
[7783-00-8]Colorless or white crystals, efflorescent in dry
2 mL of a freshly prepared 1 in 5 solution of salicylaldehyde
air and hygroscopic in moist air. Soluble in water and in
in isopropyl alcohol, mix, and allow to stand until a yellow
alcohol.
precipitate is formed. Extract the mixture with two 15-mL
AssayAccurately weigh about 100 mg, transfer to a
portions of methylene chloride. Combine the methylene
glass-stoppered flask, and dissolve in 50 mL of water. Add
chloride extracts, and dry over anhydrous sodium sulfate.
10 mL of potassium iodide solution (3 in 10) and 5 mL of
Decant the methylene chloride solution, and evaporate it to
hydrochloric acid, mix, insert the stopper in the flask, and
dryness. Recrystallize the residue of salicylaldazine from a
allow to stand for 10 minutes. Dilute with 50 mL of water,
mixture of warm toluene and methanol (60:40) with cool-
add 3 mL of starch TS, and titrate with 0.1 N sodium thio-
ing. Filter, and dry the crystals in vacuum.
sulfate VS until the color is no longer diminished, then ti-
trate with 0.1 N iodine VS to a blue color. Subtract the
volume of 0.1 N iodine solution from the volume of 0.1 N
ceed 1.
sodium thiosulfate to give the volume of 0.1 N thiosulfate
ChromatographyProceed as directed in Limit of hydra-
equivalent to selenious acid. Each mL of 0.1 N sodium thio-
zine under Povidone: the chromatogram shows only one
sulfate is equivalent to 3.225 mg of H2SeO3: not less than
spot.
93% is found.
Salicylaldehyde, (2-Hydroxybenzaldehyde),
Insoluble matterDissolve 1 g in 5 mL of water: the solu-
2-HOC6H4CHO122.12 [90-02-8]Clear, colorless to
tion is clear and complete.
yellowish-green liquid. Specific gravity: about 1.17. Slightly
soluble in water; soluble in alcohol and in ether. May con-
1.0 mg (0.01%), from 10 g.
tain a stabilizer.
Selenate and sulfateDissolve 500 mg in 10 mL of water,
AssayWhen examined by gas-liquid chromatography,
and add 0.1 mL of hydrochloric acid and 1 mL of barium
using suitable apparatus and conditions, it shows a purity of
chloride TS: no turbidity or precipitate is formed within
not less than 98%.
10 minutes.
Sand, Standard 20- to 30-MeshSilica sand, composed
Selenium, SeAt. Wt. 78.96 [7782-49-2]Dark-red
almost entirely of naturally rounded grains of nearly pure
amorphous, or bluish-black, crystalline powder. Soluble in
quartz. Predominantly graded to pass an 850-m (No. 20)
solutions of sodium and potassium hydroxides or sulfides;
sieve (85 to 100 percentage passing) and be retained on a
600-m (No. 30) sieve (0 to 5 percentage passing).
insoluble in water. Use a suitable grade with a content of Silica MicrospheresUse a suitable grade.
not less than 99.99%. [NOTEA suitable grade, in a controlled-diameter, spheri-
Selenomethionine, C5H11NO2Se196.11 cal, porous form, is available commercially as Zorbax Sil,
[1464-42-2][CAUTIONHandle with care, as this reagent is from Agilent, www.agilent.com.]
highly toxic.] Siliceous Earth, Chromatographic
For gas chromatography, use a specially prepared grade
AssayWeigh accurately about 750 mg, dissolve in
meeting the following general description: Purified siliceous
100 mL of methanol, add crystal violet TS, and titrate with
earth of suitable mesh size that has been acid- and/or base-
0.1 N perchloric acid to a blue-green endpoint. Each mL of
washed. It may or may not be silanized.
0.1 N perchloric acid is equivalent to 19.61 mg of
For column partition chromatography, it is essential that
C5H11NO2Se: between 97.0% and 103.0%, calculated on
the material be free from interfering substances. If such in-
the as-is basis, is found.
terferences are known or thought to be present, purify the
Melting range 741: about 260, with decomposition.
material as follows: Place a pledget of glass wool in the base
Nitrogen content 461Determine by the Kjeldahl
of a chromatographic column having a diameter of 100 mm
method: between 6.8% and 7.4%, calculated on the as-is
or larger, and add Purified Siliceous Earth (NF monograph) to
basis, is found.
a height equal to 5 times the diameter of the column. Add
Silica, Calcined DiatomaceousSee Diatomaceous Silica,
a volume of hydrochloric acid equivalent to one-third the
Calcined.
volume of siliceous earth, and allow the acid to percolate
Silica, Chromatographic, Silanized, Flux-Calcined, Acid-
into the column. Wash the column with methanol, using
washedUse a suitable grade.
small volumes at first to rinse the walls of the column, and
[NOTESuitable grades are available commercially as Aer-
continue washing with methanol until the last washing is
opak 30, Diatoport S, and Gas-Chrom Z.]
neutral to moistened litmus paper. Extrude the washed col-
Silica GelAn amorphous, partly hydrated SiO2 occurring
umn into shallow dishes, heat on a steam bath to remove
in glassy granules of varying size. When used as a desiccant,
the excess methanol, and dry at 105 until the material is
it frequently is coated with a substance that changes color
powdery and free from traces of methanol. Store the dried
when the capacity to absorb water is exhausted. Such
material in well-closed containers.
colored products may be regenerated (i.e., may regain their
[NOTEA suitable grade is Chromosorb W-AW.]
capacity to absorb water) by being heated at 110 until the
[NOTESuitable silanized grades for gas chromatography
gel assumes the original color.
are Gas Chrom Q, and Chromosorb W (AW- DMCS-
For use as a desiccant, use ACS Silica Gel Desiccant grade.
treated).]
Silica Gel, Binder-FreeSilica gel for chromatographic
[NOTEA suitable grade for column chromatography is
use formulated without a binder, since only activated forms
acid-washed Celite 545, available from Sigma-Aldrich,
of the silica gel are used as the binding agent.
[NOTEA suitable grade is available commercially as Sil-
Siliceous Earth, Chromatographic, SilanizedPlace
ica Gel H.]
about 450 g of purified siliceous earth in a large, open, glass
Silica Gel, ChromatographicUse a suitable grade.
crystallizing dish in a vacuum desiccator containing 30 mL
[NOTEA suitable grade is available commercially as Sil-
of a suitable silane, e.g., a mixture of 1 volume of
ica Gel G.]
dimethyldichlorosilane and 1 volume of trimethylchlorosi-
Silica Gel, Octadecylsilanized ChromatographicUse a
lane, or a mixture of 1 volume of methyltrichlorosilane and
suitable grade.
2 volumes of dimethyldichlorosilane. Apply vacuum inter-
[NOTEA suitable grade is available commercially as Re-
mittently for several hours, until no liquid silane remains.
versed Phase Uniplates from Analtech, www.analtech.com.]
Float the treated purified siliceous earth on water, and gen-
Silica Gel, PorousUse a grade suitable for high-pressure
tly agitate to allow any uncoated particles to sink. Skim the
liquid chromatography.
silanized material off the surface, wash it on a sintered-glass
[NOTEA suitable grade for reverse phase high-pressure
funnel with warm methanol until the filtrate no longer is
liquid chromatography is available as LiChrosorb SI60, Re-
acidic, and dry at 110.
verse Phase.]
Silicic Acid, SiO2 xH2O(anhydrous) 60.08
Silica GelImpregnated Glass Microfiber SheetUse a
[1343-98-2]White, amorphous powder. Insoluble in water
suitable grade.
and in acids; soluble in hot solutions of strong alkalies.
Residue on ignition (Reagent test): not less than 80.0%.
Seprachrom Chamber with Type SG ITLC, Product No.
Nonvolatile with hydrofluoric acidHeat 500 mg with
51923, from Gelman Instrument Co., Ann Arbor, MI
1 mL of sulfuric acid and 10 mL of hydrofluoric acid in a
48106.]
platinum crucible to dryness, and ignite to constant weight:
Silica Gel Mixture, ChromatographicA mixture of silica
the weight of the residue does not exceed 1.0 mg (0.2%).
gel with a suitable fluorescing substance.
[NOTEA suitable grade is available commercially as Sil- Chloride (Reagent test)One g shows not more than
ica Gel GF 254.] 0.05 mg of Cl (0.005%).
Silica Gel Mixture, Chromatographic, with Chemically
Sulfate (Reagent test)Boil 2 g with 20 mL of dilute hy-
Bound Amino GroupsUse a suitable grade.
drochloric acid (1 in 40), filter, neutralize the filtrate with
Silica Gel Mixture, Dimethylsilanized, Chromato-
ammonia TS, and dilute with water to 20.0 mL. A 10-mL
graphicUse a suitable grade.
aliquot of the solution shows not more than 0.1 mg of SO4
[NOTEA suitable grade is available as Silica Gel 60 si-
(0.01%).
lanized RP-2 F254, from EMD Chemicals,
Heavy metals (Reagent test)Boil 2.5 g with 50 mL of di-
www.emdchemicals.com.]
lute hydrochloric acid (1 in 10) for 5 minutes, filter while
Silica Gel Mixture, Octadecylsilanized Chromato-
hot, and evaporate the filtrate on a steam bath to dryness.
graphicUse a suitable grade.
Take up the residue in 20 mL of dilute hydrochloric acid (1
[NOTEA suitable grade is available commercially as KC-
in 500), digest for 5 minutes, cool, add water to make
18F from Whatman Chemical Separation, Inc., 9 Bridewell
100 mL, and filter. To 40 mL of the filtrate add 10 mL of
Place, Clifton, NJ 07014.]
hydrogen sulfide TS: any color produced is not darker than
Silica Gel Mixture, Octylsilanized, Chromatographic
that produced by adding 10 mL of hydrogen sulfide TS to a
Use a mixture of RP-8 chromatographic silica gel with a suit-
control containing 0.03 mg of Pb (0.003%).
able fluorescing substance agent.
Iron 241To 20 mL of the filtrate obtained in the test
for Heavy metals add 1 mL of hydrochloric acid, and dilute
with water to 47 mL: the solution shows not more than titrate with 0.02 N hydrochloric acid VS to the disappear-
0.015 mg of Fe (0.003%). ance of any pink color: not more than 0.20 mL is required
Silicic AcidImpregnated Glass Microfilament Sheets (0.016% as NaOH).
with Fluorescent IndicatorUse a suitable grade. -Sitosterol, (22:23 Dihydrostigmasterol), C29H50O
[NOTEOne example of a suitable grade is ITLC Type 414.7 [83-46-5]White powder. Soluble in chloroform.
SAF sheets, available from Gelman Instrument Co., 600 Store in a freezer.
South Wagner Rd., Ann Arbor, MI 48106.]
Specific rotation 781S: between 33 and 39, deter-
Silicon Carbide, SiC40.10 [409-21-2]In small
mined in a solution containing 0.5 g of test specimen per
clean chips, suitable for use in promoting ebullition.
mL of chloroform.
Silicone (75 Percent Phenyl, Methyl)Use a suitable
Water, Method I 921: not more than 6%.
grade.
Soda LimeUse Soda Lime (NF monograph).
[NOTEA suitable grade is available as OV-25.]
Sodium, NaAt. Wt. 22.98977 [7440-23-5]Use
Silicotungstic Acid, n-Hydrate (Tungstosilicic Acid),
ACS reagent grade.
H4Si(W3O10)4 nH2O2878.17 (anhydrous)
Sodium Acetate, NaC2H3O2 3H2O136.08
[12520-88-6]Green powder.
[6131-90-4]Use ACS reagent grade Sodium Acetate Trihy-
AssayDissolve about 1 g, accurately weighed, in 25 mL
drate.
of dilute hydrochloric acid (1 in 5). Add 50 mL of a solution
Sodium Acetate, Anhydrous, NaC2H3O282.03
of 5 g of cinchonine in dilute hydrochloric acid (1 in 2).
Warm on a steam bath for about 30 minutes. Cool, filter
Sodium Alizarinsulfonate (Alizarin Red S; Alizarin Sodium
through a tared crucible, and ignite at 800 to constant
Monosulfonate), C14H7NaO7S H2O360.27Yellow-brown
weight. The weight of the residue multiplied by 1.047 is
or orange-yellow powder. Freely soluble in water, with pro-
equal to the weight of silicotungstic acid dihydrate in the
duction of a yellow color; sparingly soluble in alcohol.
sample taken. Not less than 98% is found.
SensitivenessAdd 3 drops of a solution of it (1 in 100)
Silver Diethyldithiocarbamate, (C2H5)2NCS2Ag256.14
to 100 mL of water, and add 0.25 mL of 0.02 N sodium
hydroxide: a red color is produced. Add 0.25 mL of 0.02 N
Silver Nitrate, AgNO3169.87 [7761-88-8]Use ACS
hydrochloric acid: the original yellow color returns.
reagent grade.
Sodium Ammonium Phosphate (Microcosmic Salt),
Silver Oxide, Ag2O231.74 [20667-12-3]Brownish-
NaNH4HPO4 4H2O209.07Colorless crystals or white
black, heavy powder. Slowly decomposes on exposure to
granules. Freely soluble in water; insoluble in alcohol. Efflo-
light. Absorbs carbon dioxide when moist. Practically insolu-
resces in air and loses ammonia.
ble in water; freely soluble in dilute nitric acid and in am-
Insoluble matter and ammonium hydroxide precipitate
monia; insoluble in alcohol. Store in well-closed containers;
Dissolve 10 g in 100 mL of water, add 10 mL of ammonia
do not expose to ammonia fumes or easily oxidizable sub-
TS, and heat on a steam bath for 1 hour. If any precipitate
stances.
is formed, filter, wash well with water, and ignite: the ig-
AssayDissolve about 500 mg, previously dried at 120
nited precipitate weighs not more than 1 mg (0.01%).
for 3 hours and accurately weighed, in a mixture of 20 mL
of water and 5 mL of nitric acid. Dilute with 100 mL of
0.02 mg of Cl (0.002%).
water, add 2 mL of ferric ammonium sulfate TS, and titrate
Heavy metalsDissolve 3 g in 25 mL of water, add 15 mL with 0.1 N ammonium thiocyanate VS to a permanent red-
of 1 N sulfuric acid, then add 10 mL of hydrogen sulfide TS: dish-brown color. Each mL of 0.1 N ammonium thiocyanate
any brown color developed in 1 minute is not darker than is equivalent to 11.59 mg of Ag2O: not less than 99.7% of
that of a control containing 3 mL of Standard Lead Solution Ag2O is found.
(see 231) and 0.5 mL of 1 N sulfuric acid (0.001%).
Loss on dryingDry it at 120 for 3 hours: it loses not
NitrateDissolve 1 g in 10 mL of water, add 0.1 mL of more than 0.25% of its weight.
indigo carmine TS, then add, with stirring, 10 mL of sulfuric
NitrateTo 500 mg add 30 mg of sodium carbonate and
acid: the blue color persists for 10 minutes (about 0.005%).
2 mL of phenoldisulfonic acid TS, mix, and heat on a steam
Sulfate (Reagent test, Method II)Dissolve 10 g in bath for 15 minutes. Cool, cautiously add 20 mL of water,
100 mL of water, add 5 mL of hydrochloric acid, and filter if render alkaline with ammonia TS, and dilute with water to
necessary: the filtrate yields not more than 5 mg of residue 30 mL: any color produced by the test solution is not darker
(0.02%). than that produced in a control containing 0.01 mg of NO3
Sodium Arsenate (Arsenic Acid Sodium Salt), Na2HAsO4 (0.002%).
Substances insoluble in nitric acidDissolve 5 g in a mix-
Sodium Arsenite, NaAsO2129.91 [7784-46-5]
ture of 5 mL of nitric acid and 10 mL of water, dilute with
White, crystalline powder. Soluble in water; slightly soluble
water to about 65 mL, and filter any undissolved residue on
in alcohol.
a tared filtering crucible (retain the filtrate for the test for
AssayTransfer about 5.5 g, accurately weighed, to a Substances not precipitated by hydrochloric acid). Wash the
500-mL volumetric flask, dissolve in and dilute with water to crucible with water until the last washing shows no opales-
volume, and mix. Pipet 25 mL of this solution into a suitable cence with 1 drop of hydrochloric acid, and dry at 105 to
container, add 50 mL of water and 5 g of dibasic sodium constant weight: the residue weighs not more than 1 mg
phosphate, swirl to dissolve, and titrate with 0.1 N iodine (0.02%).
VS, adding 3 mL of starch TS as the endpoint is approached.
Substances not precipitated by hydrochloric acidDilute
Each mL of 0.1 N iodine is equivalent to 3.746 mg of As.
the filtrate obtained in the test for Substances insoluble in
Between 57.0% and 60.5% is found (equivalent to 98.8%
nitric acid with water to 250 mL, heat to boiling, and add,
to 104.9% of NaAsO2).
dropwise, sufficient hydrochloric acid to precipitate all of the
Chloride (Reagent test)One g shows not more than silver (about 5 mL), avoiding any great excess. Cool, dilute
0.10 mg of Cl (0.01%). with water to 300 mL, and allow to stand overnight. Filter,
evaporate 200 mL of the filtrate in a suitable tared porcelain Heavy metalsDissolve 200 mg in 8 mL of dilute hydro-
dish to dryness, and ignite: the residue weighs not more chloric acid (3 in 8), and evaporate on a steam bath to
than 1.7 mg (0.05%). dryness. Dissolve the residue in 5 mL of dilute hydrochloric
acid (2 in 5), and again evaporate to dryness. Dissolve the AlkalinityHeat 2 g with 40 mL of water on a steam bath
residue in 10 mL of water, and add 2 mL of diluted acetic for 15 minutes, cool, and dilute with water to 50 mL. Filter,
acid and 10 mL of hydrogen sulfide TS. Any brown color discarding the first 10 mL of the filtrate. To 25 mL of the
subsequent filtrate add 2 drops of phenolphthalein TS, and
produced is not darker than that of a control containing Iodine contentAdd 10 mL to 5 mL of toluene contained
0.01 mg of added Pb (0.005%). in a 125-mL separator fitted with a suitable inert plastic
stopcock, and shake vigorously for 2 min. Extract gently
IronDissolve 1 g in 20 mL of dilute hydrochloric acid (1
with three 10-mL portions of dilute phosphoric acid (1 in
in 5), and add, dropwise, a slight excess of bromine TS. Boil
3), combining the lower phases in a 125-mL iodine flask.
the solution to remove the excess bromine, cool, dilute with
Add sodium hypochlorite TS, dropwise, to the combined ex-
water to 40 mL, and add 10 mL of ammonium thiocyanate
tracts until the solution turns brown, then add 0.5 mL in
solution (3 in 10). Any red color produced is not darker
excess. Shake intermittently for 3 min, add 5 mL of freshly
than that of a control containing 0.02 mg of added Fe
prepared, saturated phenol solution, mix, and allow to
(0.02%).
stand for 1 min, accurately timed. Add 1 g of potassium
SulfideDissolve 1 g in 20 mL of water, and add 5 drops
iodide, shake for 30 s, add 3 mL of starch TS, and titrate
of lead acetate TS: no brown color is produced (about
with 0.1 N sodium thiosulfate VS: NMT 0.1 mL of 0.1 N
0.0005%).
sodium thiosulfate is consumed.
Sulfate (Reagent test, Method II)Dissolve 5 g in 100 mL
of water, add methyl orange TS, neutralize with 1 N hydro-
277134 from www.sigma-aldrich.com or as catalog number
chloric acid, add 3 mL of the acid in excess, and filter: the
54101 from www.gfschemicals.com.]
filtrate yields not more than 3 mg of residue (0.02%).
Sodium Biphosphate, NaH2PO4 H2O137.99Use ACS
Sodium Azide, NaN365.01 [26628-22-8]White
reagent grade Sodium Phosphate, Monobasic.
powder.
Sodium Bisulfite [7631-90-5]This reagent is usually
Assay[CAUTIONSodium azide is a potent poison. Its a mixture of sodium bisulfite and sodium metabisulfite
conjugate acid HN3 is more toxic than hydrogen cyanide [7681-57-4]. Use ACS reagent grade Sodium Bisulfite.
and is readily liberated from neutral aqueous solutions. Con- Sodium Bitartrate, NaHC4H4O6 H2O190.08
tact of NaH3 or hydrazoic acid (HN3) with certain metals [6131-98-2]White crystals or a crystalline powder. Soluble
may produce explosive salts. Work in a well-ventilated hood, in cold water.
and handle the sample with care.] Dissolve about 100 mg,
accurately weighed, in 50 mL of water, and add 3 drops of
30 mL of water, add phenolphthalein TS, and titrate with
phenolphthalein. Adjust the pH, if necessary, to 7.0, and
0.1 N sodium hydroxide VS: each mL of 0.1 N sodium hy-
add 35.0 mL of 0.1 N perchloric acid. Pipet, while stirring,
droxide is equivalent to 19.01 mg of NaHC4H4O6 H2O. Be-
2.5 mL of 1.0 M sodium nitrite into the solution, and stir for
tween 99% and 100.5% is found.
15 seconds. Titrate rapidly to the phenolphthalein endpoint
with 0.1 N sodium hydroxide. The endpoint should be
from 10 g (0.01%).
reached in less than 4 minutes after addition of perchloric
acid because HN3 is readily volatile. Calculate the percent-
0.2 mg of Cl (0.02%).
age of azide by the formula:
Heavy metals (Reagent test)Dissolve 4 g in 25 mL of
[(NP)(VP) (NS)(VS)](65.01)(100)/2C water, add 2 drops of phenolphthalein TS, and then add
ammonia TS, dropwise, until the solution is slightly pink.
where NP is the normality of perchloric acid solution; VP is Add 4 mL of 1 N hydrochloric acid, dilute with water to
the volume of perchloric acid, in mL, taken; NS is the nor- 40 mL, and add 10 mL of hydrogen sulfide TS: any brown
mality of sodium hydroxide solution; VS is the volume, in color produced is not darker than that of a control contain-
mL, of sodium hydroxide taken; 65.01 is the molecular ing 0.04 mg of added Pb (0.001%).
weight of sodium azide; and C is the weight, in mg, of
Sulfate (Reagent test, Method I )One g shows not more
sodium azide. Not less than 98.5% of NaN3 is found.
than 0.2 mg of SO4 (0.02%).
Sodium Bicarbonate, NaHCO384.01 [144-55-8]
Sodium Borate (Borax; Sodium Tetraborate), Na2B4O7
Sodium Biphenyl, C12H9Na176.19Available as a solu-
[NOTECertified Borax is available from the National Insti-
tion in 2-ethoxyethyl ether, or in 1,2-dimethoxyethane (di-
tute of Standards and Technology, Washington, DC, www.
ethylene glycol diether).
nist.gov, as standard sample No. 187.]
ActivityPlace 20 mL of dry toluene in a titration flask Sodium Borohydride, NaBH437.83 [16940-66-2]
equipped with a magnetic stirring bar and a stopper having White, crystalline solid. Freely soluble in water; soluble (with
a hole through which the delivery tip of a weight buret may reaction) in methanol. Its solutions are rapidly decomposed
be inserted. Add a quantity of sodium biphenyl sufficient to by boiling.
produce a blue color in the mixture, and titrate with amyl
Assay
alcohol, contained in a weight buret, to the disappearance
POTASSIUM IODATE SOLUTION (0.25 N)Dissolve 8.917 g, pre-
of the blue color. (Disregard the amounts of sodium bi-
viously dried at 110 to constant weight and accurately
phenyl and amyl alcohol used in this adjustment.) Weigh
weighed, in water to make 1000.0 mL.
accurately the weight buret containing the amyl alcohol.
PROCEDUREDissolve about 500 mg, accurately weighed,
Transfer the contents of a vial of well-mixed test specimen
in 125 mL of sodium hydroxide solution (1 in 25) in a
to the titration flask, and titrate quickly with the amyl alco-
250-mL volumetric flask, dilute with the sodium hydroxide
hol to the disappearance of the blue color. Weigh the buret
solution to volume, and mix. Pipet 10 mL of the solution
to determine the weight of amyl alcohol consumed, and
into a 250-mL iodine flask, add 35.0 mL of Potassium iodate
calculate the activity, in mEq/vial, by the formula:
solution, and mix. Add 2 g of potassium iodide, mix, add
10 mL of dilute sulfuric acid (1 in 10), insert the stopper in Result = 11.25W
the flask, and allow to stand in the dark for 3 minutes. Ti-
trate the solution with 0.1 N sodium thiosulfate VS, adding in which W is the weight of amyl alcohol consumed. NLT
3 mL of starch TS as the endpoint is approached. Calculate 10% activity is found.
the amount, in mg, of NaBH4 in the specimen titrated by
the formula: Sodium Diethyldithiocarbamate, (C2H5)2NCS2Na
([(35.0)(0.25)] 0.1V)4.729 Sodium 2,2-dimethyl-2-silapentane-5-sulfonateSee So-
dium 3-(trimethylsilyl)-1-propane sulfonate.
in which V is the volume, in mL, of 0.1 N sodium thiosulfate Sodium DithioniteUse Sodium Hydrosulfite.
used in the titration. Not less than 98% is found. Sodium Dodecyl Sulfate (Sodium Lauryl Sulfate),
Sodium Bromide, NaBr102.89 [7647-15-6]Use C12H25SO4Na288.38 [151-21-3]Light yellow, crystal-
ACS reagent grade. line powder.
Sodium CarbonateUse Sodium Carbonate, Anhydrous. Sodium Ferrocyanide, Na4Fe(CN)6 10H2O484.06
Sodium Carbonate, Anhydrous, Na2CO3105.99 [13601-19-9]Yellow crystals or granules. Freely soluble in
[497-19-8]Use ACS reagent grade. water.
Sodium Carbonate, Monohydrate, Na2CO3 H2O
AssayDissolve 2 g, accurately weighed, in 400 mL of
water, add 10 mL of sulfuric acid, and titrate with 0.1 N
Sodium Chloride, NaCl58.44 [7647-14-5]Use ACS
potassium permanganate VS. Each mL of 0.1 N potassium
reagent grade.
permanganate is equivalent to 48.41 mg of Na4Fe(CN)6
Sodium Chloride Solution, IsotonicUse Saline TS.
10H2O. Not less than 98% is found.
Sodium Cholate Hydrate (3,7,12-trihydroxy-5-cho-
lan-24-oic acid sodium salt; cholalic acid sodium salt),
from 10 g (0.01%).
C24H39NaO5 xH2O430.55 (anhydrous) [206986-87-0]
Chloride (Reagent test)Dissolve 1 g in 75 mL of water,
It can be from ox or sheep bile. Use a suitable grade with a
add a solution prepared by dissolving 1.2 g of cupric sulfate
in 25 mL of water, mix, and allow to stand for 15 minutes.
Sodium Chromate, Na2CrO4 4H2O234.03
To 20 mL of the decanted, clear liquid add 2 mL of nitric
[7775-11-3]Lemon-yellow crystals. Soluble in water.
acid and 1 mL of silver nitrate TS: any turbidity produced
AssayAccurately weigh about 300 mg, and dissolve in
does not exceed that of a control containing 0.02 mg of Cl,
10 mL of water contained in a 500-mL flask. Add 3 g of
2 mL of nitric acid, 1 mL of silver nitrate TS, and sufficient
potassium iodide and 10 mL of diluted sulfuric acid, and
cupric sulfate to match the color of the Test solution.
dilute with 350 mL of oxygen-free and carbon dioxide-free
SulfateDissolve 5 g in 100 mL of water without heating,
water. Titrate the liberated iodine with 0.1 N sodium thio-
filter, and to the filtrate add 0.25 mL of glacial acetic acid
sulfate VS, adding 3 mL of starch TS as the endpoint is ap-
and 5 mL of barium chloride TS: no turbidity is produced in
proached. Each mL of 0.1 N sodium thiosulfate consumed is
10 minutes (about 0.01% as SO4).
equivalent to 7.802 mg of Na2CrO4 4H2O. Not less than
Sodium Fluorescein, C20H10Na2O5376.28Orange-red,
99% is found.
hygroscopic powder. Freely soluble in water; slightly soluble
in alcohol. Its water solution is yellowish red in color and
from 20 g dissolved in 150 mL of water (0.005%).
exhibits a strong yellowish green fluorescence that disap-
AluminumDissolve 20 g in 140 mL of water, filter, and
pears when the solution is acidified and reappears when the
add 5 mL of glacial acetic acid to the filtrate. Add stronger
solution is neutralized or made basic.
ammonia water until alkaline, and digest for 2 hours on a
steam bath. Pass through hardened filter paper, wash thor-
oughly, ignite, and weigh: the residue weighs not more
Sodium Fluoride, NaF41.99 [7681-49-4]Use ACS
than 0.8 mg (0.002%).
reagent grade.
CalciumDetermine as directed in the test for calcium
Sodium Glycocholate, C26H42NNaO6487.60
for ACS reagent grade Potassium Chromate (0.005%).
[863-57-0]White to tan powder. Is hygroscopic. Freely sol-
ChlorideDetermine as directed in the test for chloride uble in water and in alcohol.
for ACS reagent grade Potassium Chromate (about
Specific rotation 781: between +28 and +31, calcu-
0.005%).
lated on the dried basis (it is rendered anhydrous by drying
SulfateDetermine as directed in the test for sulfate for at 100 for 2 hours), determined at 20 in a solution con-
ACS reagent grade Potassium Dichromate, but add 4.5 mL taining 10 mg per mL.
of hydrochloric acid to the water used to dissolve the test
Nitrogen, Method I 461: between 2.6% and 3.2% of
specimen: the residue weighs not more than 2.4 mg
N is found, calculated on the dried basis.
(0.01%).
Sodium 1-Heptanesulfonate (1-Heptanesulfonic Acid
Sodium ChromotropateSee Chromotropic Acid.
Sodium Salt), C7H15NaO3S202.25 [22767-50-6]Use a
Sodium Citrate Dihydrate (2-Hydroxy-1,2,3-propanetri-
suitable grade.
carboxylic Acid, Trisodium Salt, Dihydrate), Na3C6H5O7
Sodium 1-Heptanesulfonate Monohydrate,
C7H15NaO3S H2O220.26 [22767-50-6]Use a suitable
Sodium Cobaltinitrite, Na3Co(NO2)6403.94
grade.
Sodium 1-Hexanesulfonate (1-Hexanesulfonic Acid Sodium
Sodium Cyanide, NaCN49.01 [1433-33-9]Use
Salt), C6H13NaO3S188.22 [2832-45-3]Use a suitable
ACS reagent grade.
grade.
Sodium 1-Decanesulfonate (1-Decanesulfonic Acid Sodium
Sodium 1-Hexanesulfonate Monohydrate,
Salt), C10H21NaO3S244.33 [13419-61-9]Use a suitable
C6H13NaO3S H2O206.23 [2832-45-3]Use a suitable
grade for ion pair chromatography with a content of not
grade.
less than 99.0%.
Sodium Hydrogen Sulfate (Sodium Bisulfate), NaHSO4
Sodium DesoxycholateUse Bile Salts.
120.06 [7681-38-1]Freely soluble in water; very soluble
Sodium Dichromate, Na2Cr2O7 2H2O (for chromic acid
in boiling water. It decomposes in alcohol into sodium sul-
cleaning mixture)298.00 [7789-12-0]Use ACS rea-
fate and free sulfuric acid. Use a suitable reagent grade.
gent grade.
Sodium Hydrosulfite (Sodium Dithionite), Na2S2O4
174.11 [7775-14-6]White or grayish-white crystalline
powder. Soluble in water; slightly soluble in alcohol. Gradu-
ally oxidizes in air, more readily when in solution, to bisul-
fite, acquiring an acid reaction. Is affected by light.
AssayAccurately weigh about 1 g, dissolve it in a mix- dryness. Take up the residue in 20 mL of water, and filter if
ture of 10 mL of formaldehyde TS and 10 mL of water con- necessary. To the filtrate add ammonium hydroxide until the
tained in a small glass-stoppered flask, and allow to stand solution is just alkaline, then add 4 drops of ammonium hy-
for 30 minutes with frequent agitation. Transfer the solution droxide and 5 mL of ammonium oxalate TS: any turbidity
to a 250-mL volumetric flask, add 150 mL of water and produced within 15 minutes does not exceed that in a blank
3 drops of methyl orange TS, and then add, dropwise, 1 N containing 0.1 mg of added Ca carried through the entire
sulfuric acid to a slightly acid reaction. Dilute with water to procedure (0.001%).
250 mL, and mix. To 50.0 mL of the dilution add 2 drops of
Phosphate (Reagent test)Transfer 2 g to a beaker, and
phenolphthalein TS and just sufficient 0.1 N sodium hydrox-
add 5 mL of hydrochloric acid and 2 g of potassium iodide.
ide to produce a slight, pink color, then titrate with 0.1 N
Heat the solution for 5 minutes, and cool. Add 2 mL of
iodine, adding 3 mL of starch TS as the indicator. Then dis-
30 percent hydrogen peroxide, and evaporate the solution
charge the blue color of the solution with 1 drop of 0.1 N
to dryness. Rinse the walls of the beaker with a few mL of
sodium thiosulfate, and titrate with 0.1 N sodium hydroxide
water, and add 2 mL of hydrochloric acid and 2 mL of
VS to a pink color: each mL of 0.1 N sodium hydroxide is
30 percent hydrogen peroxide. Evaporate again to dryness:
equivalent to 3.482 mg of Na2S2O4. Not less than 88% is
the residue shows not more than 0.01 mg of PO4 (5 ppm).
found.
Sodium Iodate, NaIO3197.9 [7681-55-2]White to
SulfideAdd sodium hydroxide solution (1 in 10) to lead yellowish-white powder. Use a suitable reagent grade.
acetate TS until the precipitate dissolves. Add 5 drops of this Sodium Lauryl SulfateSee Sodium Dodecyl Sulfate.
solution to a solution of 1 g of the sodium hydrosulfite in Sodium Metabisulfite, Na2S2O5190.11
10 mL of water: no immediate darkening is observed. [7681-57-4]Use ACS reagent grade.
Sodium Metaperiodate, NaIO4213.89
Heavy metalsDissolve 1 g in 10 mL of water, add 10 mL
[7790-28-5]Use ACS reagent grade Sodium Periodate.
of hydrochloric acid, and evaporate on a steam bath to dry-
Sodium Methoxide, CH3ONa54.02 [124-41-4]
ness. Dissolve the residue in 20 mL of water and 0.5 mL of
Fine, white powder. Reacts violently with water with evolu-
diluted hydrochloric acid, filter, and add to the filtrate
tion of heat. Soluble in alcohol and in methanol.
10 mL of hydrogen sulfide TS: no darkening is produced.
Render the solution alkaline with ammonia TS: a slight, AssayTransfer about 220 mg to a tared, glass-stoppered
greenish color may be produced, but not a dark or white flask, and weigh accurately. Dissolve the test specimen in
precipitate. about 10 mL of methanol, then add 100 mL of water slowly,
with stirring. Add phenolphthalein TS, and titrate with 0.1
Suitability for riboflavin assayTo each of 2 or more tubes
N hydrochloric acid VS to a colorless endpoint: each mL of
add 10 mL of water and 1.0 mL of a standard riboflavin so-
0.1 N hydrochloric acid VS is equivalent to 5.402 mg of
lution containing 20 g of riboflavin in each mL, and mix.
CH3ONa. Not less than 98.0% is found.
To each tube add 1.0 mL of glacial acetic acid, mix, add
Sodium Molybdate, Na2MoO4 2H2O241.95
with mixing, 0.5 mL of potassium permanganate solution (1
in 25), and allow to stand for 2 minutes. Then to each tube
Sodium Nitrate, NaNO384.99 [7631-99-4]Use
add, with mixing, 0.5 mL of hydrogen peroxide TS: the per-
ACS reagent grade.
manganate color is destroyed within 10 seconds. Shake the
Sodium Nitrite, NaNO269.00 [7632-00-0]Use ACS
tubes vigorously until excess oxygen is expelled. If gas bub-
reagent grade.
bles remain on the sides of tubes after foaming has ceased,
Sodium Nitroferricyanide (Sodium Nitroprusside), Na2Fe
remove the bubbles by tipping the tubes so that the solu-
(NO)(CN)5 2H2O297.95 [13755-38-9]Use ACS rea-
tion flows slowly from end to end. In a suitable fluorometer,
gent grade.
measure the fluorescence of the solution. Then add, with
Sodium 1-OctanesulfonateSee Octanesulfonic Acid So-
mixing, 8.0 mg of sodium hydrosulfite: the riboflavin is
dium Salt.
completely reduced in not more than 5 seconds.
Sodium Oxalate, Na2C2O4134.00 [62-76-0]Use
Sodium Hydroxide, NaOH40.00 [1310-73-2]Use
ACS reagent grade.
ACS reagent grade.
[NOTESodium Oxalate of a quality suitable as a primary
Sodium Hypochlorite Solution [7681-52-9]A solution
standard is available from the Office of Standard Reference
of sodium hypochlorite (NaOCl) in water. Usually yellow to
Materials, National Institute of Standards and Technology,
yellowish-green in color. Has an odor of chlorine. Is affected
Washington, DC, www.nist.gov, as standard sample No.
by light and gradually deteriorates. Store it in light-resistant
40.]
containers, preferably below 25. [CAUTIONThis solution is
Sodium (tri) Pentacyanoamino Ferrate [Trisodium
corrosive and may evolve gases that are corrosive and toxic.
Aminepentacyanoferrate (3-)], Na3[Fe(CN)5NH3]271.93
It is a powerful oxidant that can react violently with reduc-
[14099-05-9]Yellow to tan powder. Soluble in water.
ing agents. Is irritating and corrosive to skin and mucous
membranes.] SolubilityDissolve 500 mg in 50 mL of water, and allow
to stand for 1 hour: the solution is clear and free from for-
AssayTransfer about 3 mL to a tared, glass-stoppered
eign matter.
iodine flask, and weigh accurately. Add 50 mL of water, 2 g
of potassium iodide, and 10 mL of acetic acid, insert the Sensitivity
stopper in the flask, and allow to stand in the dark for 1,1-DIMETHYLHYDRAZINE STANDARD SOLUTIONPlace 500 mL of
10 minutes. Remove the stopper, rinse the walls of the flask water in a 1-L volumetric flask, and add from a buret
with a few mL of water, and titrate the liberated iodine with 1.27 mL of anhydrous 1,1-dimethylhydrazine. Dilute with
0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as water to volume, and mix. Pipet 10 mL of this solution into
the endpoint is neared. Each mL of 0.1 N sodium thiosulfate a 100-mL volumetric flask, and dilute with water to volume.
consumed is equivalent to 3.723 mg of NaOCl: not less Each mL of this solution contains the equivalent of 100 g
than 5.25% is found. If it is desired to calculate the percent- of 1,1-dimethylhydrazine.
age of available chlorine, note that each mL of 0.1 N so-
BUFFER SOLUTIONTransfer 4.8 g of citric acid monohydrate
dium thiosulfate consumed is equivalent to 3.545 mg of
to a 1-L volumetric flask, dissolve in water, add 14.6 g of
available chlorine.
sodium phosphate, swirl to dissolve, and dilute with water
CalciumTransfer 10.0 g to a 150-mL beaker, dissolve in to volume.
10 mL of water, and add 5 mL of hydrochloric acid and 2 g
TEST PREPARATIONDissolve 100 mg of sodium (tri)pentacy-
of potassium iodide. Heat the mixture for 5 minutes, cool,
anoamino ferrate in 100 mL of water.
and add 2 mL of 30 percent hydrogen peroxide. Evaporate
PROCEDUREInto each of five 25-mL volumetric flasks pi-
to dryness, cool, and add 2 mL of hydrochloric acid and
pet 0 mL, 0.25 mL, 0.50 mL, 1.0 mL, and 1.5 mL, respec-
2 mL of 30 percent hydrogen peroxide. Rinse the inner walls
tively, of 1,1-Dimethylhydrazine standard solution; to each
of the beaker with a few mL of water, and evaporate to
add 15 mL of Buffer solution,and swirl to mix. To each flask, glass electrode and a calomel electrode modified to use 0.1
add by pipet 2 mL of Test preparation, mix, dilute with Buffer N tetramethylammonium chloride in methanol as the elec-
solution to volume, and allow to stand for 1 hour. Using a trolyte. Perform a blank determination, and make any neces-
suitable spectrophotometer, 1-cm cells, and the solution sary correction. Each mL of 0.1 N perchloric acid is equiva-
containing no 1,1-Dimethylhydrazine standard solution as the lent to 11.00 mg of CH3COCO2Na: not less than 98.0% is
blank, determine the absorbances of the remaining solutions found.
at 500 nm. Plot the observed absorbance as the ordinate
SolubilityDissolve 1.5 g in 25 mL of water: the solution
versus the concentration of standard as the abscissa on co-
is clear and complete.
ordinate paper, and draw the curve of best fit. The plot is
Free acidDissolve 10 g in 150 mL of water, and titrate
linear and the absorbance of the 150-g solution is not less
with 0.5 N sodium hydroxide VS, determining the endpoint
than 0.65.
potentiometrically: not more than 2.8 mL of 0.5 N sodium
Sodium 1-Pentanesulfonate (1-Pentanesulfonic Acid So-
hydroxide is consumed (about 1% as C3H4O3).
dium Salt), C5H11NaO3S H2O192.21 [207605-40-1]
Sodium Salicylate [54-21-7]It complies with the specifi-
Use a suitable grade with a content of NLT 98.0%.
cations under Sodium Salicylate (USP monograph), and in
Sodium 1-Pentanesulfonate, Anhydrous
addition meets the requirements of the following test.
(1-Pentanesulfonic Acid Sodium Salt, Anhydrous),
NitrateDissolve 100 mg in 5 mL of water, and superim-
C5H11SO3Na174.19 [22767-49-3]Use a suitable grade
pose the solution upon 5 mL of sulfuric acid: no brownish-
with a content of NLT 98%.
red color appears at the junction of the two liquids.
Sodium Perchlorate, NaClO4 H2O140.46
Sodium Selenite, Na2SeO3172.94 [10102-18-8]
White, odorless, crystalline powder, usually partially hy-
Sodium Peroxide, Na2O277.98 [1313-60-6]Use
drated. Freely soluble in water; insoluble in alcohol.
ACS reagent grade.
AssayAccurately weigh about 180 mg, previously dried Sodium Phosphate, Dibasic (Disodium Phosphate; Diso-
at 120 to constant weight, and dissolve it in 50 mL of dium Hydrogen Phosphate; Sodium Phosphate, Dibasic,
water in a glass-stoppered flask. Add, successively, 3 g of Heptahydrate), Na2HPO4 7H2O268.07 [7782-85-6]
potassium iodide and then 5 mL of hydrochloric acid, insert Use ACS reagent grade Sodium Phosphate, Dibasic,
the stopper, and allow to stand for 10 minutes. Add 50 mL Heptahydrate.
of water, 50.0 mL of 0.1 N sodium thiosulfate VS, and 3 mL Sodium Phosphate, Dibasic, Anhydrous (Anhydrous Di-
of starch TS, and immediately titrate with 0.1 N iodine VS sodium Hydrogen Phosphate) (for buffer solutions), Na2HPO4
to a blue color. Perform a blank determination. The differ- 141.96 [7558-79-4]Use ACS reagent grade Sodium
ence in volumes of 0.1 N iodine is equivalent to 4.323 mg Phosphate, Dibasic, Anhydrous.
of Na2SeO3. Between 98% and 101% is found. Sodium Phosphate, Dibasic, Dihydrate (Sodium
Monohydrogen Phosphate; Disodium Hydrogen Phosphate), SolubilityOne g in 10 mL of water shows not more than
Na2HPO4 2H2O177.99 [10028-24-7]Use a suitable a faint haze.
grade with a content of not less than 99.5%. [NOTEA suit-
CarbonateTo 500 mg add 1 mL of water and 2 mL of
able grade is available from www.emdchemicals.com, cata-
diluted hydrochloric acid: no effervescence is produced.
log number SX0713.]
Chloride (Reagent test)A 500-mg portion shows not
Sodium Phosphate Dibasic, Dodecahydrate (Disodium
more than 0.05 mg of Cl (0.01%).
Hydrogen Phosphate, Dodecahydrate), Na2HPO4.12H2O
Nitrate (Reagent test)A 200-mg portion dissolved in
358.14 [10039-32-4]Use a suitable grade with a con-
3 mL of water shows not more than 0.02 mg of NO3
tent of between 98.0% and 102.0% of Na2HPO4 12H2O.
(0.01%).
Sodium Phosphate, Dibasic, Heptahydrate (Disodium
Hydrogen Phosphate Heptahydrate; Disodium Phosphate), Selenate and sulfate (as SO4)To 500 mg in a small evap-
Na2HPO4 7H2O268.07 [7782-85-6]Use ACS reagent orating dish add 20 mg of sodium carbonate and 10 mL of
grade. hydrochloric acid. Slowly evaporate the solution on a steam
Sodium Phosphate, Monobasic (Sodium Biphosphate; So- bath under a hood to dryness. Wash the sides of the dish
dium Dihydrogen Phosphate; Acid Sodium Phosphate; with 5 mL of hydrochloric acid, and again evaporate to dry-
Monosodium Orthophosphate), NaH2PO4 H2O137.99 ness. Dissolve the residue in a mixture of 15 mL of hot
[10049-21-5]Use ACS reagent grade. water and 1 mL of hydrochloric acid. Proceed as directed
Sodium Phosphate, Monobasic, Anhydrous (Sodium under Sulfate in Reagents (Reagent test, Method I), begin-
Biphosphate; Sodium Dihydrogen Phosphate; Acid Sodium ning with Filter the solution. The test specimen shows no
Phosphate; Monosodium Orthophosphate), NaH2PO4119.98 more turbidity than that produced by 0.15 mg of SO4
[7558-80-7]Use a suitable grade with a content of not less (0.03%).
than 99.0%. Sodium Sulfate (Glaubers Salt), Na2SO4 10H2O322.20
Sodium Phosphate, Monobasic, Dihydrate (Sodium [7727-73-3]Use ACS reagent grade.
Dihydrogen Phosphate, Dihydrate), NaH2PO4 2H2O156.01 Sodium Sulfate, Anhydrous, Na2SO4142.04
[13472-35-0]Use a suitable grade with a content of not [7757-82-6]Use ACS reagent grade.
less than 99.0%. For use in assaying alkaloids by gasliquid chromatogra-
Sodium Phosphate, Tribasic, Na3PO4 12H2O380.12 phy, it conforms to the following additional test.
Suitability for alkaloid assaysTransfer about 10 mg of at-
Sodium Phosphite Pentahydrate (Disodium Hydrogen
ropine, accurately weighed, to a 25-mL volumetric flask, dis-
Phosphite), Na2HPO3 5H2O216.04 [13517-23-2]Use a
solve in and dilute with alcohol to volume. Pipet 3 mL of
the solution into each of two 60-mL separators, and add to
[NOTEA suitable grade is available from Fluka, www.
each 10 mL of water, 1 mL of 1 N sodium hydroxide, and
sigma-aldrich.com, catalog number 04283.]
10 mL of chloroform. Shake thoroughly, and allow the lay-
Sodium Pyrophosphate, Na4P2O7 10H2O446.06
ers to separate. Filter the organic phase from one separator
through phase-separating paper, previously washed with
Sodium Pyruvate, CH3COCO2Na110.04
5 mL of chloroform, supported in a funnel, and collect the
[113-24-6]White to practically white powder or crystalline
filtrate in a suitable container. Add 10 mL of chloroform to
solid. Soluble in water.
the separator, shake thoroughly, and filter the organic layer
AssayTransfer about 300 mg, accurately weighed, to a through the same phase-separating paper, collecting and
high-form titration beaker, add 150 mL of glacial acetic acid, combining the filtrates in the same container. Designate the
and stir until dissolved. Titrate with 0.1 N perchloric acid combined filtrates as Solution A. Filter the organic phase
VS, determining the endpoint potentiometrically, using a from the second separator through 30 g of the Anhydrous
Sodium Sulfate, supported on a pledget of glass wool in a Sodium L-ThyroxineUse Levothyroxine Sodium (USP
small funnel, and previously washed with chloroform, and monograph).
collect the filtrate in a suitable container. Add 10 mL of Sodium 3-(trimethylsilyl)-1-propane sulfonate (Sodium
chloroform to the separator, shake thoroughly, and filter the 2,2-dimethyl-2-silapentane-5-sulfonate), C6H15SiNaO3S
organic layer through the same portion of anhydrous so- 218.32Use a suitable grade.
dium sulfate, collecting and combining the two filtrates in Sodium Tungstate, Na2WO4 2H2O329.85
the same container. Designate the combined filtrates as So- [10213-10-2]Use ACS reagent grade.
lution B. Evaporate the two solutions in vacuum to a volume Soluble StarchSee Starch, Soluble.
of about 1 mL. Inject an accurately measured volume of So- Solvent HexaneSee Hexane, Solvent.
lution A into a suitable gas chromatograph, and record the SorbitolUse Sorbitol (NF monograph).
peak height. Repeat the determination with a second accu- Stachyose Hydrate, C24H42O21 xH2O666.58
rately measured volume of Solution A, record the peak [10094-58-3]Use a suitable grade.
height, and obtain the average of the two results. In a simi- Standard Sand, 20- to 30-MeshSee Sand, Standard 20-
lar manner, determine the peak height of two portions of to 30-mesh.
Solution B, and obtain the average of the results. The aver- Stannous Chloride, SnCl2 2H2O225.65
age value obtained for Solution B is within 5.0% of the value [10025-69-1]Use ACS reagent grade.
obtained for Solution A. Starch, PotatoThe starch separated from the tubers of
Under typical conditions, the gas chromatograph contains Solanum tuberosum Linn e (Fam. Solanaceae). A more or less
a 4-mm 1.2-m glass column packed with 3% phase G3 finely granular powder, consisting of starch grains of charac-
on packing S1A. After curing and conditioning, the column teristic shape and appearance when examined microscopi-
temperature is maintained at 210, the injector port temper- cally.
ature at 225, and the detector block temperature at 240 Starch, Soluble (for iodimetry) [9005-84-9]Use ACS re-
during the determinations. The carrier gas is helium, flowing agent grade.
at a rate of 60 mL per minute. Starch, Soluble, PurifiedWhite, amorphous powder;
Sodium Sulfate DecahydrateUse Sodium Sulfate. under microscopic examination it shows the characteristic
Sodium Sulfide, Na2S 9H2O240.18 [1313-84-4] form of potato starch. Soluble in hot water; very slightly
Use ACS reagent grade. soluble in alcohol.
Sodium SulfiteUse Sodium Sulfite, Anhydrous.
TEST SOLUTION FOR DETERMINATION OF PH AND SENSITIVITYStir
Sodium Sulfite, Anhydrous (Exsiccated Sodium Sulfite),
2.0 g in 10 mL of water, add boiling water to make 100 mL,
Na2SO3126.04 [7753-83-7]Use ACS reagent grade.
and boil for 2 minutes. The hot solution is almost clear. On
Sodium p-Sulfophenylazochromotropate [Trisodium Salt
cooling, the solution may become opalescent or turbid, but
of 4,5-Dihydroxy-3-(p-sulfophenylazo)-2,
does not gel. Use it as the Test solution.
7-naphthalenedisulfonic Acid], C16H9N2Na3O11S3 3H2O
pH 791The pH of the Test solution is between 6.0 and
624.47Bright red powder. Very soluble in water; insoluble
7.5.
in alcohol. Combines with zirconium oxychloride to form a
SensitivityMix 2.5 mL of Test solution, 97.5 mL of water,
soluble pink zirconium lake.
and 0.50 mL of 0.010 N iodine: a distinct blue color results,
[NOTEThe reagent is available as Catalog No. 7309 from
and it disappears upon the addition of 0.50 mL of 0.010 N
Distillation Products Industries, Eastman Organic Chemicals
sodium thiosulfate.
Dept., Rochester, NY 14650. A procedure for its preparation
AbsorbancePrepare a pH 5.3 buffer solution by dissolv- is described in Z. Anal. Chem., 146, 417 (1955).]
ing 43.5 g of sodium acetate (trihydrate) and 4.5 mL of gla- Sodium Tartrate, Na2C4H4O6 2H2O230.08
cial acetic acid in water, transferring the resultant solution [6106-24-7]Use ACS reagent grade.
to a 250-mL volumetric flask, adding water to volume, and Sodium Tetraphenylborate, NaB(C6H5)4342.22
mixing. [143-66-8]Use ACS reagent grade.
Dissolve 1.00 g of Soluble Purified Starch in 2.5 mL of the Sodium TetraphenylboronSee Sodium Tetraphenylbo-
buffer solution by warming, transfer to a 100-mL volumetric rate.
flask, add water to volume, and mix. Add 0.50 mL of this Sodium Thioglycolate (Sodium Thioglycollate),
solution to a 100-mL volumetric flask containing about HSCH2COONa114.10 [367-51-1]A white, crystalline
75 mL of water, 1 mL of 1 N hydrochloric acid, and 1.5 mL powder. Very soluble in water; slightly soluble in alcohol. Is
of 0.020 N iodine, swirling the flask during the addition. hygroscopic, and oxidizes in air. Store in tight, light-resistant
Add water to volume, mix, and allow to stand in the dark containers. It should not be used if it is pale yellow or
for 1 hour. The absorbance of this solution, measured at darker in color.
575 nm in a 1-cm cell against a blank, is between 0.5 and
AssayAccurately weigh about 250 mg, and dissolve in
0.6.
50 mL of oxygen-free water. Add 5 mL of diluted hydrochlo-
Reducing substancesShake 10.0 g with 100 mL of water ric acid, boil for 2 minutes, cool, and titrate the solution
for 15 minutes, and allow to settle for about 12 hours. Filter with 0.1 N iodine VS, adding 3 mL of starch TS toward the
a portion of the supernatant through fine sintered glass. To end: each mL of 0.1 N iodine is equivalent to 11.41 mg of
50 mL of the filtrate add 50 mL of alkaline cupric tartrate TS, HSCH2COONa. Not less than 75% is found.
and boil for 1 to 2 minutes. Filter the resulting cuprous ox-
Insoluble matterA solution of 1 g in 10 mL of water is
ide, wash it with hot water and then with alcohol, and dry
clear, and practically complete.
it at 105 for 2 hours: not more than 47 mg is found, corre-
SulfideDissolve 500 mg in 10 mL of water in a small
sponding to 0.7% of reducing sugars as maltose.
flask, add 2 mL of hydrochloric acid, then place a strip of
filter paper, moistened with lead acetate TS, over the mouth
of the flask, and bring the solution to a boil: the lead ace-
Residue on ignition 281: not more than 0.5%. tate paper is not darkened.
Stearic Acid, C18H36O2284.48 [57-11-4]Hard, Sodium Thiosulfate, Na2S2O3 5H2O248.19
white crystals or amorphous, white powder. Freely soluble [10102-17-7]Use ACS reagent grade.
in chloroform and in ether; soluble in alcohol and in solvent
hexane.
Congealing temperature 651: between 67 and 69. Assay and carbonateAccurately weigh about 5 g, dis-
solve in 200 mL of warm carbon dioxide-free water in a
Acid value 401: between 196 and 199.
glass-stoppered, 500-mL flask, add phenolphthalein TS, and
Iodine value 401: not more than 1.
titrate with 1 N hydrochloric acid VS to determine the hy-
Saponification value 401: between 197 and 200.
droxide alkalinity. Then add methyl orange TS, and titrate
Palmitic acidDetermine as directed in the Assay under with 1 N hydrochloric acid VS. Each mL of 1 N hydrochloric
Stearic Acid (NF monograph): not more than 5.0% is found. acid required to reach the phenolphthalein endpoint is
Stearyl Alcohol (1-Octadecanol), C18H38O270.49 equivalent to 132.9 mg of Sr(OH)2 8H2O, and each addi-
[112-92-5]White flakes, granules, or crystals. Soluble in al- tional mL of 1 N hydrochloric acid VS required to reach the
cohol, in ether, in acetone, and in benzene; insoluble in methyl orange endpoint is equivalent to 73.8 mg of SrCO3.
water. Not less than 95.0% of Sr(OH)2 8H2O and not more than
3.0% of SrCO3 are found. Melting range 741: between 56 and 58.
Chloride (Reagent test)Dissolve 1.0 g in 100 mL of Other requirementsIt conforms to the tests for Acid
water, and filter if necessary: 1.0 mL of the solution shows value, Iodine value, and Hydroxyl value under Stearyl Alcohol
not more than 0.01 mg of Cl (0.1%). (NF monograph).
Stronger Ammonia WaterSee Ammonia Water, Calcium (Reagent test)
Stronger.
TEST SOLUTIONDissolve 5.0 g in water, and dilute with
Strontium Acetate, Sr(CH3COO)2
1
/2H2O214.72
water to 100 mL.
[543-94-2]White, crystalline powder. Soluble in 3 parts of
SAMPLE SOLUTIONDilute 10.0 mL of the Test solution with
water; slightly soluble in alcohol.
water to 100 mL.
AssayIgnite about 3 g, accurately weighed, in a plati-
CONTROL SOLUTIONTo 10.0 mL of the Test solution add
num crucible, protecting from sulfur in the flame. Cool,
0.50 mg of calcium ion (Ca), and dilute with water to
transfer the crucible with the residue to a beaker, and add
100 mL.
50 mL of water and 40.0 mL of 1 N hydrochloric acid VS.
PROCEDUREDetermine the background emission at
Boil gently for 30 minutes or longer, if necessary; filter; wash
416.7 nm: the limit is 0.1%.
with hot water until the washings are neutral; add methyl
red TS; and titrate the excess acid with 1 N sodium hydrox- IronDissolve 1 g in warm water, and dilute with water
ide VS. Each mL of 1 N hydrochloric acid is equivalent to to 100 mL. To 20 mL of this solution add 2 mL of hydro-
107.4 mg of Sr(CH3COO)2
1
/2H2O: not less than 99% is chloric acid and 0.1 mL of 0.1 N potassium permanganate,
found. allow to stand for 5 minutes, and add 3 mL of ammonium
thiocyanate solution (3 in 10). Any red color produced is
not darker than that of a control containing 0.03 mg of
from 10 g (0.02%).
added Fe (0.015%).
Free alkali or free acidDissolve 3 g in 30 mL of water,
Heavy metalsDissolve 2.0 g in 14 mL of dilute hydro-
and add 3 drops of phenolphthalein TS: no pink color is
chloric acid (1 in 6), and evaporate on a steam bath to
produced. Titrate with 0.1 N sodium hydroxide VS to a pink
dryness. Take up the residue in 25 mL of water, filter, and
color: not more than 0.30 mL of the 0.1 N sodium hydrox-
dilute with water to 100 mL (Test solution). To 5.0 mL of the
ide is required.
Test solution add 0.02 mg of lead (Pb), and dilute with water
BariumDissolve 1 g in 10 mL of water, and add 1 drop
to 30 mL, to provide the standard. For the test specimen,
of glacial acetic acid and 5 drops of potassium dichromate
use 30 mL of the Test solution. Adjust each solution with
solution (1 in 10): no turbidity is produced within 2 minutes
diluted acetic acid or ammonia TS to a pH between 3.0 and
(about 0.02%).
4.0 (using short-range pH paper), dilute with water to
CalciumIgnite 1 g until completely carbonized. Warm
40 mL, and add 10 mL of freshly prepared hydrogen sulfide
the residue with a mixture of 3 mL of nitric acid and 10 mL
TS: any brown color developed in the sample solution is not
of water, filter, wash with 5 mL of water, and evaporate the
darker than that in the control solution (0.004%).
filtrate on a steam bath to dryness. Powder the residue, and
Strychnine Sulfate, (C21H22N2O2)2 H2SO4 5H2O856.98
dry it at 120 for 3 hours. Reflux the dried powder with
[60-41-3]Colorless or white crystals, or a white, crystalline
15 mL of dehydrated alcohol for 10 minutes, cool in ice,
powder. Its solutions are levorotatory. One g dissolves in
and filter. Repeat the extraction with 10 mL of dehydrated
about 35 parts of water, in 85 mL of alcohol, and in about
alcohol. Evaporate the combined filtrates to dryness, add
220 mL of chloroform. Insoluble in ether.
0.5 mL of sulfuric acid, and ignite: the weight of the residue
SolubilityA solution of 500 mg in 25 mL of water is
is not more than 10 mg (0.3% of Ca).
complete, clear, and colorless.
0.1 mg of Cl (0.01%).
0.1%.
Heavy metals (Reagent test): 0.001%.
BrucineTo 100 mg add 1 mL of dilute nitric acid (1 in
Iron 241Dissolve 1.0 g in 45 mL of water, and add
2): a yellow color may be observed, but not a red or red-
2 mL of hydrochloric acid: the solution shows not more
dish-brown color.
than 0.01 mg of Fe (0.001%).
Styrene-Divinylbenzene Anion-exchange Resin, 50- to
Alkali saltsDissolve 2 g in 80 mL of water, heat to boil- 100-MeshSee Anion-exchange Resin, 50- to 100-Mesh, Sty-
ing, add an excess of ammonium carbonate TS, boil for rene-Divinylbenzene.
5 minutes, dilute with water to 100 mL, and filter. Evaporate Styrene-Divinylbenzene Cation-exchange Resin,
50 mL of the filtrate, and ignite: the residue, after correcting Strongly AcidicSee Cation-exchange Resin, Styrene-
for the ignition residue from half the volume of the clear Divinylbenzene, Strongly Acidic.
ammonium carbonate TS used above, is not more than Styrene-Divinylbenzene Copolymer BeadsNeutral, po-
3 mg (0.3%). rous, cross-linked beads, 200400 mesh, molecular weight
operating range up to 2,000 (based on beads fully swollen NitrateDissolve 1 g in 10 mL of water, add 0.10 mL of
in benzene). Suitable for use in the gel permeation separa- indigo carmine TS, and then add 10 mL of sulfuric acid: the
tion of lipophilic polymers and other solutes requiring or- blue color persists for 5 minutes (about 0.01% of NO3).
ganic eluant. Strontium Hydroxide (Strontium Hydroxide Octahydrate),
Sr(OH)2 8H2O265.76 [18480-07-4]White, crystalline,
free-flowing powder. Sparingly soluble in water. May absorb
carbon dioxide from the air. Keep tightly closed.
[NOTEA suitable grade is available commercially as Bi- cence does not exceed that of quinine sulfate solution (1 in
oBeads S-X from Bio-Rad, www.bio-rad.com.] 1,600,000,000), similarly measured.
Succinic Acid, C4H6O4118.09 [771-50-6]Use ACS Sulfuric Acid, Fuming, H2SO4 plus free SO3
reagent grade. [8014-95-7]having a nominal content of 15%, 20%, or
Sudan III, C22H16N4O352.39 [85-86-9]Red to red- 30% of free SO3Use ACS reagent grade (containing be-
brown powder. Use a suitable grade. tween 15.0% and 18.0%, between 20.0% and 23.0%, or
between 30.0% and 33.0% of free SO3).
AssayWhen tested by thin-layer chromatography (see
Sulfuric Acid, Nitrogen Free, H2SO498.08
Chromatography 621) with the use of plates coated with
[NOTEA suitable grade is available as Sulfuric Acid, Ultrex
consisting of a mixture of hexane and ethyl acetate (80:20),
II, catalog number 6902-05 from www.mallbaker.com.]
and examined under short-wavelength UV light, a single
Sulfurous Acid, H2SO382.08 [7782-99-2]A water
spot is exhibited, with trace impurities.
solution of sulfur dioxide. Use ACS reagent grade.
Sudan IV, C24H20N4O380.44 [85-83-6]Brown to
Sunflower Oil [8001-21-6]Use a suitable grade.
reddish-brown powder.
Supports for Gas ChromatographySee supports for
gas chromatography in the Chromatographic Reagents sec-
100-mL volumetric flask. Dissolve in chloroform, dilute with
tion under Chromatography 621.
chloroform to volume, and mix. Dilute 2.0 mL of the result-
Tannic Acid (Tannin) [1401-55-4]Use ACS reagent
ing solution with chloroform to 50.0 mL. Determine the ab-
grade.
sorbance of this solution in 1-cm cells at the wavelength of
Tartaric Acid, H2C4H4O6150.09Use ACS reagent
maximum absorbance at about 520 nm, with a suitable
grade.
spectrophotometer, using chloroform as the blank. Calculate
Tertiary Butyl AlcoholSee Butyl Alcohol, Tertiary.
the percentage of Sudan IV in the test specimen taken by
Testosterone Benzoate, C26H32O2376.53Use a suita-
the formula:
ble grade.
2,4,5,7-Tetrabromofluorescein (Eosin Y, Eosin Yellowish,
(100A)/(85C)
Eosin Bromo ES, Solvent Red 43, Acid Red 87), C20H8Br4O5
167.8 [630-20-6]Dark red to brown powder. Solubility
in which A is the absorbance at 520 nm and C is the con-
0.1% in water (clear orange solution). Use a suitable grade.
centration of the test specimen in g per L. Not less than
Tetrabromophenolphthalein Ethyl Ester, C22H14Br4O4
90% is found.
Tetrabutylammonium Bromide, (C4H9)4NBr322.37
Sulfamerazine (4-Amino-N-(4-methyl-2-pyrimidinyl)-
Tetrabutylammonium Hydrogen Sulfate, C16H37NO4S
benzenesulfonamide), C11H12N4O2S264.30 [127-79-7]
339.54 [32503-27-8]White, crystalline powder. Soluble
Use a suitable grade with a content of NLT 99.0%.
in alcohol yielding a slightly hazy, colorless solution.
Sulfamic Acid, HSO3NH297.09 [5329-14-6]Use
ACS reagent grade.
40 mL of water. Titrate with 0.1 N sodium hydroxide VS,
Sulfanilamide, C6H8N2O2S172.21 [63-74-1]Use
determining the endpoint potentiometrically. Perform a
USP Sulfanilamide Melting Point RS.
blank determination and make any necessary correction.
Sulfanilic Acid, p-NH2C6H4SO3H H2O191.21
33.95 mg of C16H37NO4S. Not less than 97.0% is found.
Sulfatase Enzyme PreparationUse a suitable grade.
Melting range 741: between 169 and 173. [NOTEA suitable grade is available commercially under
Tetrabutylammonium Hydrogen Sulfate Ion Pairing Re- catalog number S-9626 from Sigma-Aldrich, Web site: www.
agentA mixture of tetrabutylammonium hydrogen sulfate sigma-aldrich.com.]
and phosphate buffer. Use a suitable grade. Sulfathiazole Sodium (4-Amino-N-2-thiazolylbenzenesul-
[NOTEA suitable grade is available as catalog number fonamide Sodium Salt), C9H8N3NaO2S2277.29
WAT084189 from the Waters Corporation (www.waters. [144-74-1]Use a suitable grade.
com).] Sulfonic Acid Cation-exchange ResinSee Cation-ex-
0.4 M Aqueous Tetrabutylammonium Hydroxide, change Resin, Sulfonic Acid.
C16H37NO259.47 [2052-49-5]Use a suitable grade. Sulfosalicylic Acid, C6H3(COOH)(OH)(SO3H)-1,2,5
[NOTEA suitable grade is available as catalog number 2H2O254.22 [97-05-2]Use ACS reagent grade.
420120025 from www.fishersci.com.] SulfurUse Precipitated Sulfur (USP monograph).
Tetrabutylammonium Hydroxide, 1.0 M in Methanol Sulfur Dioxide Detector TubeA fuse-sealed glass tube
[2052-49-5]Use a suitable grade. so designed that gas may be passed through it and contain-
Tetrabutylammonium Hydroxide 30-Hydrate, ing suitable absorbing filters and support media for an io-
C16H37NO 30H2O799.93 [2052-49-5]Use a suitable dine-starch indicator.
Tetrabutylammonium Hydroxide, 40 Percent in Water,
[CH3(CH2)3]4NOH259.47 [2052-49-5]Use a suitable
grade.
Tetrabutylammonium Iodide, (C4H9)4NI369.37
Sulfuric Acid, H2SO498.08 [7664-93-9]Use ACS re-
[311-28-4]White, shiny, crystalline flakes. Soluble in alco-
agent grade.
hol and in ether; slightly soluble in water.
Sulfuric Acid, Diluted (10 percent)Cautiously add
AssayDissolve 370 mg, accurately weighed, in 60 mL of 57 mL of sulfuric acid to about 100 mL of water, cool to
acetone with vigorous stirring. Stir the solution by mechani- room temperature, and dilute with water to 1000 mL.
cal means, add 10 mL of 16% sulfuric acid, and titrate with Sulfuric Acid, FluorometricUse ACS reagent grade Sul-
0.1 N silver nitrate VS, determining the endpoint potentio- furic Acid that conforms to the following additional test:
metrically, using a glasssilver electrode system, and adding
FluorescenceUsing a suitable fluorometer having a sharp
the titrant in 0.1-mL increments as the endpoint is ap-
cut-off 360-nm excitation filter and a sharp cut-off 415-nm
proached. Perform a blank determination, and make any
excitation filter, determine the fluorescence of the sulfuric
necessary corrections. Each mL of 0.1 N silver nitrate is
acid in a cuvette previously rinsed with water followed by
several portions of the acid under examination: the fluores-
equivalent to 36.94 mg of (C4H9)4NI: not less than 99.0% is Boiling range (Reagent test): between 177 and 187,
found. at a pressure of 9 mm of Hg.
Tetrabutylammonium Phosphate, (C4H9)4NH2PO4 Tetraethylenepentamine, C8H23N5189.31
339.46 [5574-97-0]White to off-white powder. Soluble [112-57-2]Colorless liquid.
in water.
AssayDissolve about 1.5 g, accurately weighed, in ograph (see Chromatography 621) equipped with a flame-
100 mL of water. Without delay, titrate with 0.5 N sodium ionization detector, helium being used as the carrier gas.
hydroxide VS, determining the endpoint potentiometrically. The following conditions have been found suitable: a 0.25-
Perform a blank determination, and make any necessary mm 30-m capillary column coated with a 1-m layer of
correction. Each mL of 0.5 N sodium hydroxide is equiva- phase G2; the injection port temperature is maintained at
lent to 169.7 mg of (C4H9)4NH2PO4. Not less than 97.0% is 250; the detector temperature is maintained at 300; and
found. the column temperature is maintained at 150 and pro-
2,3,7,8-Tetrachlorodibenzo-p-dioxin,
13
C-labeled, grammed to rise 10 per minute to 280. The area of the
13
C12H4Cl4O2333.84Clear, colorless liquid. C8H23N5 peak is not less than 30% of the total peak area.
AssayInject an appropriate volume into a gas chromat- Refractive index 831: between 1.503 and 1.507 at
ograph (see Chromatography 621) equipped with an elec- 20.
tron-capture detector, helium being used as the carrier gas. Tetraheptylammonium Bromide, (C7H15)4NBr490.70
The following conditions have been found suitable: a 0.25- [4368-51-8]White, flaky powder.
Tetrahexylammonium Hydrogen Sulfate, C24H53NO4S
451.75 [32503-34-7]Use a suitable grade with a con-
tent of not less than 98.0%.
Delta-8-tetrahydrocannabinol (-
13
C12H4Cl4O2 peak is not less than 99.0% of the total peak
8-Tetrahydrocannabinol), C21H30O2314.47 [5957-75-5]
area.
Use a suitable grade which may be a solid material or a
[NOTEA suitable grade is available from Cambridge Iso-
solution in methanol.[NOTEA suitable grade of a methano-
topes Laboratories (www.isotope.com).]
lic solution (1 mg/mL) is available from www.cerilliant.com,
2,3,7,8-Tetrachlorodibenzofuran,
13
C-labeled,
catalog number T-032.]
13
C12H4Cl4O317.84Clear, colorless liquid.
Tetrahydrofuran, C4H8O72.11 [109-99-9]Use ACS
AssayInject an appropriate volume into a gas chromat- reagent grade.
ograph (see Chromatography 621) equipped with an elec- Tetrahydrofuran, Peroxide-Free, C4H8O72.11Use
tron-capture detector, helium being used as the carrier gas. ACS reagent grade.
PeroxideTransfer 8 mL of potassium iodide and starch
TS to a 12-mL ground glass-stoppered cylinder about
15 mm in diameter. Fill completely with the substance
under test, mix, and allow to stand protected from light for
5 minutes. No color develops.
Tetrahydrofuran, Stabilizer-FreeUse a suitable grade.
13
C12H4Cl4O peak is not less than 99.0% of the total peak
Tetrahydro-2-furancarboxylic Acid (Tetrahydro-2-furoic
area.
Acid), C5H8O3116.12 [16874-33-2]Use a suitable
[NOTEA suitable grade is available from Cambridge Iso-
grade with a content of not less than 97%. [NOTEA suita-
topes Laboratories (www.isotope.com).]
ble grade is available from www.sigma-aldrich.com, catalog
1,1,2,2-Tetrachloroethane, C2H2Cl4167.8
number 341517.]
[630-20-6]Colorless clear liquid. Specific gravity: 1.553.
N-(2-Tetrahydrofuroyl)piperazine (1-[(Tetrahydro-
Refractive index at 20 is 1.481. Use a suitable grade.
2-furanyl)carbonyl]piperazine), C9H16N2O2184.23Use a
Tetracosane, C24H50338.66 [646-31-1]White pow-
suitable grade.
der.
[NOTEA suitable grade can be obtained from EMS-
Melting range 741: between 51 and 53. DOTTIKON, www.ems-dottikon.ch.]
Tetradecane, C14H30198.39 [629-59-4]Clear, col- 1,2,3,4-Tetrahydronaphthalene, C10H12132.21
orless liquid. [119-64-2]Colorless liquid.
AssayWhen examined by gas-liquid chromatography, it Refractive index 831: 1.5401 at 20.
shows a purity of not less than 98%. The following condi- Tetramethylammonium Bromide, (CH3)4NBr154.05
tions have been found suitable for assaying the reagent: a [64-20-0]Use ACS reagent grade.
3-mm 2.4-m stainless steel column packed with phase Tetramethylammonium Chloride, (CH3)4NCl109.60
G16 on support S1; the carrier gas is helium, flowing at a [75-57-0]Colorless crystals. Soluble in water and in alco-
rate of 27.5 mL per minute; the column temperature is hol; insoluble in chloroform.
maintained at 250, the injection port is maintained at
200, and the detector is maintained at 280. A flame-ioni-
beaker, add 50 mL of water and 10 mL of diluted nitric acid,
zation detector is employed.
swirl to dissolve the test specimen, add 50.0 mL of 0.1 N
Melting range, Class II 741: between 4 and 8, within silver nitrate VS, and mix. Add 2 mL of ferric ammonium
a 2 range. sulfate TS and 5 mL of nitrobenzene, shake, and titrate the
excess silver nitrate with 0.1 N ammonium thiocyanate VS:
each mL of 0.1 N silver nitrate is equivalent to 10.96 mg of
20.
(CH3)4NCl. Not less than 98% is found.
Tetraethylammonium Perchlorate, (C2H5)4NClO4
Tetramethylammonium Hydroxide, (CH3)4NOH91.15
229.70White crystals. Soluble in water. Use a suitable
[75-59-2]Available as an approximately 10% or approxi-
grade.
mately 25% aqueous solution, or as the crystalline pentahy-
Tetraethylene Glycol, C8H18O5194.23 [112-60-7]
drate. Is clear and colorless. Tetramethylammonium hydrox-
Nearly colorless liquid. Refractive index: about 1.46.
ide is a stronger base than ammonia and rapidly absorbs
AssayWhen examined by gasliquid chromatography,
carbon dioxide from the air. Store in tight containers.
using suitable gas chromatographic apparatus and condi-
AssayAccurately weigh a glass-stoppered flask contain-
tions, it shows a purity of not less than 90%.
ing about 15 mL of water. Add a quantity of a solution of
tetramethylammonium hydroxide, equivalent to about
200 mg of (CH3)4NOH, and again weigh. Add methyl red acid. Heat to 60 and maintain this temperature while titrat-
TS, and titrate the solution with 0.1 N hydrochloric acid VS: ing with 0.1 N ceric sulfate VS, determining the endpoint
each mL of 0.1 N hydrochloric acid is equivalent to potentiometrically, using silver-silver chloride and platinum
9.115 mg of (CH3)4NOH. electrodes. Each mL of 0.1 N ceric sulfate is equivalent to
11.99 mg of TlCl. Not less than 99% is found.
Residue on evaporationEvaporate 5 mL of solution on a
Theobromine, C7H8N4O2180.17 [83-67-0]White,
steam bath, and dry at 105 for 1 hour: the weight of the
crystalline solid. Very slightly soluble in water and in alcohol;
residue is equivalent to not more than 0.02% of the weight
almost insoluble in benzene, in ether, and in chloroform.
of the test specimen.
Ammonia and other aminesAccurately weigh a quantity
50 mL of glacial acetic acid. Titrate with 0.1 N perchloric
of solution, corresponding to about 300 mg of (CH3)4NOH,
acid VS, determining the endpoint potentiometrically. Per-
in a low-form weighing bottle tared with 5 mL of water.
Add a slight excess of 1 N hydrochloric acid (about 4 mL),
evaporate on a steam bath to dryness, and dry at 105 for
18.02 mg of C7H8N4O2. Not less than 95% is found.
2 hours: the weight of the tetramethylammonium chloride
Thiazole Yellow (CI Direct Yellow 9; Clayton Yellow; Titan
so obtained, multiplied by 0.8317, represents the quantity,
Yellow), C28H19N5Na2O6S4695.74 [1829-00-1]Yellow-
in mg, of (CH3)4NOH in the portion of test specimen taken
ish-brown powder. Soluble in water and in alcohol to yield
and corresponds to within 0.2% above or below that found
in each instance a yellow solution; soluble in dilute alkali to
in the Assay.
yield a brownish-red solution. Protect from light.
Tetramethylammonium Hydroxide, Pentahydrate,
(CH3)4NOH 5H2O181.23 [10424-65-4]White to off- SolubilityA 200-mg portion mixed with 50 mL of water
white crystals. Is hygroscopic. Strong base. Keep well-closed. shows not more than a faint haze.
Soluble in water and in methanol.
Residue on ignitionAccurately weigh about 1.5 g, previ-
AssayAccurately weigh about 800 mg, dissolve in ously dried at 105 for 2 hours, and ignite until thoroughly
100 mL of water, and titrate with 0.1 N hydrochloric acid charred. Cool, add 2 mL of nitric acid and 2 mL of sulfuric
VS, determining the endpoint potentiometrically. Perform a acid, ignite gently to expel excess acids, then at 600 to
blank determination, and make any necessary correction. 800 to constant weight: the residue of sodium sulfate
Each mL of 0.1 N hydrochloric acid is equivalent to (Na2SO4) is between 19.8% and 21.5% of the weight of the
18.22 mg of (CH3)4NOH 5H2O: not less than 98% is found. test specimen (theory is 20.4%).
Tetramethylammonium Hydroxide Solution in Metha-
Sensitiveness to magnesiumAdd 0.2 mL of a solution (1
nol [75-59-2]A solution in methanol of tetramethylammo-
in 10,000) and 2 mL of 1 N sodium hydroxide to a mixture
nium hydroxide [(CH3)4NOH91.15]. Is generally available
of 9.5 mL of water and 0.5 mL of a solution prepared by
in concentrations of 10% and 25%. The following specifica-
dissolving 1.014 g of clear crystals of magnesium sulfate in
tions apply specifically to the 25% concentration; for other
water, diluting with water to 100 mL, then diluting 10 mL
concentrations, appropriate adjustments in the procedures
of the resulting solution with water to 1 L: a distinct pink
may be necessary.
color is produced within 10 minutes.
AssayAccurately weigh about 1 g of the solution, and Thioacetamide, C2H5NS75.13 [62-55-5]Use ACS
dilute with water to about 50 mL. Add phenolphthalein TS, reagent grade.
and titrate with 0.1 N hydrochloric acid VS to the disap- 2-Thiobarbituric Acid, C4H4N2O2S144.15
pearance of the pink color: each mL of 0.1 N hydrochloric [504-17-6]White leaflets. Slightly soluble in water.
acid VS is equivalent to 91.15 mg of (CH3)4NOH. Between
Melting temperature 741: 236, with decomposition.
23% and 25% is found.
2,2-Thiodiethanol, (HOCH2CH2)2S122.19
ClarityA portion of it in a test tube is clear, or only [111-48-8]Pale yellow to colorless liquid.
slightly turbid, when viewed transversely.
AssayNot less than 98% of C4H10O2S is found, a suita-
Tetramethylammonium Nitrate, (CH3)4NNO3136.15
ble gas chromatograph equipped with a flame-ionization
[1941-24-8]White crystals. Freely soluble in water.
detector being used. The following conditions have been
Tetramethylbenzidine, (4-(4-Amino-3,5-dimethylphenyl)-
found suitable: a 4.0-mm 1.83-m glass column is packed
2,6-dimethylaniline; 3,3,5,5 Tetramethylbenzidine; 3,35,5-
with 10% phase G25 on support S1A; the column, injection
Tetramethyl-[1,1-biphenyl] 4,4-diamine) C16H20N2240.34
port, and detector temperatures are maintained at 200,
250, and 310, respectively.
4,4-Tetramethyldiaminodiphenylmethane [(4,4-
Refractive index 831: between 1.4250 and 1.4270,
Methylenebis(N,N-dimethylaniline)], [(CH3)2NC6H4]2CH2
at 20.
254.38 [101-61-1]Off-white crystals.
Thioglycolic Acid, HSCH2COOH92.12 [68-11-1]A
colorless or nearly colorless liquid. Miscible with water. Solu-
Tetramethylsilane, (CH3)4Si88.23 [75-76-3]Use
ble in alcohol.
ACS reagent grade.
SensitivenessMix 1 mL with 2 mL of stronger ammonia
Tetrapropylammonium Chloride, C12H28ClN221.82
water, and dilute with water to 20 mL. Add 1 mL of this
solution to a mixture of 20 mL of water and 0.1 mL of dilute
than 98.0%.
ferric chloride TS (1 in 100), then add 5 mL of ammonia TS:
Tetrasodium Ethylenediaminetetraacetate
a distinct pink color is produced.
((Ethylenedinitrilo)tetraacetic Acid Tetrasodium Salt),
Thionine Acetate, C12H9N3S C2H4O2287.34
C10H12N2Na4O8380.17Fine, white, crystalline powder.
Soluble in water.
Thiourea, (NH2)2CS76.12 [62-56-6]Use ACS rea-
gent grade.
Thorium Nitrate, Th(NO3)4 4H2O552.12
Thallous Chloride, TlCl239.84Fine, white, crystalline
powder. Soluble in about 260 parts of cold water and in
Thrombin Human (Factor IIa )~33,600
about 70 parts of boiling water; insoluble in alcohol. Poison-
[9002-04-4]A preparation of a serine protease (enzyme)
ous; use with adequate ventilation.
that converts human fibrinogen into fibrin. It is obtained
AssayDissolve about 500 mg, accurately weighed, in a from human plasma and may be prepared by precipitation
mixture of 80 mL of water and 0.5 mL of sulfuric acid. with suitable salts and organic solvents under controlled
When dissolution is complete, add 20 mL of hydrochloric
conditions of pH, ionic strength, and temperature. A yellow- o-Tolidine (4,4-Diamino-3,3-dimethylbiphenyl), (NH2)
ish-white powder, freely soluble in a 9 g per L solution of (CH3)C6H3 C6H3(CH3)(NH2)4,3,3,4212.29
sodium chloride, which forms a cloudy, pale yellow solution. [119-93-7]White to reddish crystals or crystalline powder.
Store in a sealed, sterile container under nitrogen, protected Slightly soluble in water; soluble in alcohol, in ether, and in
from light, at a temperature below 0. One unit corresponds dilute acids. Preserve in well-closed containers, protected
to the amount of enzyme that hydrolyzes 1 mol of Tos- from light. [CAUTIONAvoid contact with o-tolidine and mix-
Gly-Pro-Arg-4-nitroaniline acetate per minute at a pH of 8.4 tures containing o-tolidine, and conduct all tests in a well-ven-
and a temperature of 37. tilated fume hood.]
Thromboplastin [9035-58-9]Buff-colored powder, or
opalescent or turbid suspension. It exhibits thrombokinase
Tolualdehyde (o-Tolualdehyde), C8H8O120.15
activity derived from the acetone-extracted brain and/or
lung tissue of freshly killed rabbits. It may contain sodium
p-Tolualdehyde, C8H8O120.15 [104-87-0]Color-
chloride and calcium chloride in suitable proportions, and it
less to yellow, clear liquid.
may contain a suitable antimicrobial agent. It is used in sus-
pension form for the determination of the prothrombin time
and activity of blood. Its thrombokinase activity is such that
tions have been found suitable for assaying the article: A
it gives a clotting time of 11 to 16 seconds with normal
3-mm 1.8-m stainless steel column packed with a 5%
human plasma and the proper concentration of calcium
phase G4 on support S1. Nitrogen, having a flow rate of
ions. Store in tight containers, preferably at a temperature
about 12 mL per minute, is the carrier gas, the detector and
below 5.
column temperature are about 125, and the injection port
Loss on drying 731[NOTEThis test is applicable only
temperature is about 205. A flame-ionization detector is
to the dry form.] Dry it in vacuum at 60 for 6 hours: it
employed and the specimen is a 5% solution in carbon di-
sulfide.
Thymidine, C10H14N2O5242.2 [50-89-5]White
powder. Use a suitable grade.
20.
Thymol, C6H3[CH3][OH][CH(CH3)2]1,3,4150.22
Toluene (Toluol), C6H5CH392.14 [108-88-3]Use
[89-83-8]Colorless, often large, crystals, or a white, crys-
ACS reagent grade.
talline powder. Is affected by light. Has greater density than
p-Toluenesulfonic Acid, CH3C6H4SO3H H2O190.22
water, but when liquefied by fusion is less dense than water.
Its alcohol solutions are neutral to litmus. One g dissolves in
p-Toluenesulfonyl-L-arginine Methyl Ester Hydrochlo-
about 1000 mL of water, in 1 mL of alcohol, in 1 mL of
ride (N-p-Tosyl-L-arginine methyl ester hydrochloride; TAME),
chloroform, in 1.5 mL of ether, and in about 2 mL of olive
C14H22N4O4S HCl378.88 [1784-03-8]Determine its
oil. Soluble in glacial acetic acid and in fixed or volatile oils.
suitability as directed in the test for Trypsin under Chymo-
Store in tight, light-resistant containers.
trypsin (USP monograph).
Melting range 741: between 48 and 51, but when
p-Toluic Acid, CH3C6H4COOH136.15 [99-94-5]
melted it remains liquid at a considerably lower tempera-
White, crystalline powder. Sparingly soluble in hot water;
ture.
very soluble in alcohol, in methanol, and in ether.
Nonvolatile matterVolatilize 2 g on a steam bath, and
dry at 105 to constant weight: the residue weighs not
suitable container, dissolve in 125 mL of alcohol, add 25 mL
of water, and mix. Titrate with 0.5 N sodium hydroxide VS,
Thyroglobulin, [9010-34-8]A protein having a molecu-
determining the endpoint potentiometrically. Perform a
lar weight of 670 kDa. Available as a slightly beige, freeze-
dried powder made from bovine or porcine thyroid gland.
68.07 mg of C8H8O2: not less than 98% is found.
Tin, SnAt. Wt. 118.71 [7440-31-5]Use ACS rea-
Melting range 741: over a range of 2 that includes
gent grade.
181.
Titanium Tetrachloride, TiCl4189.68 [7550-45-0]
o-Toluidine (2-Aminotoluene; 2-Methylaniline),
Clear, colorless liquid. Fumes in air. [CAUTIONIt reacts vio-
C6H4(CH3)(NH2)1,2107.15 [95-53-4]Light yellow liq-
lently with water.]
uid becoming reddish brown on exposure to air and light.
AssayAccurately weigh 0.75 g into 100 mL of 2 N sulfu-
Soluble in alcohol, in ether, and in dilute acids; slightly solu-
ric acid contained in a Smith weighing buret. Pour the solu-
ble in water. Preserve in well-closed containers, protected
tion through a zincmercury reduction column into 50 mL
from light.
of 0.1 N ferric ammonium sulfate VS. Elute with 100 mL of
Specific gravity 841: 1.008 at 20.
2 N sulfuric acid and 100 mL of water. Add 10 mL of phos-
phoric acid, and titrate with 0.1 N potassium permanganate
p-Toluidine, C7H9N107.15 [106-49-0]White to
VS. Perform a blank determination, and make any necessary
beige crystals or flakes.
correction. Each mL of 0.1 N potassium permanganate is
Freely soluble in alcohol, in acetone, in methanol, and in
equivalent to 18.97 mg of TiCl4. Not less than 99.5% is
dilute acids; slightly soluble in water.
found.
AssayDissolve 400 mg, accurately weighed, in 100 mL
of glacial acetic acid, and titrate with 0.1 N perchloric acid
Titanium Trichloride (Titanous Chloride), TiCl3154.23
VS, determining the endpoint potentiometrically. Perform a
[7705-07-9]Black, hygroscopic powder, unstable in air.
Soluble in water, the solution depositing titanic acid on ex-
Each mL of 0.1 N perchloric acid is equivalent to 10.72 mg
posure to air. Is available usually as 15% to 20%, dark vio-
of CH3C6H4NH2. Not less than 98%, calculated on the dried
let-blue, aqueous solutions. Store the solution in tightly
basis, is found.
closed, glass-stoppered bottles, protected from light.
Loss on dryingWeigh accurately about 1 g, and dry at Melting range 741: between 47 and 49.
30 to constant weight: it loses not more than 2% of its
SuitabilityDetermine its suitability for use in the test for
weight.
Related compounds under Propoxyphene Hydrochloride (USP
Toluidine Blue, (C15H16ClN3S)2 ZnCl2747.95
monograph) as follows. Dissolve a suitable quantity in chlo-
roform to yield a solution containing 20 g per mL. Follow-
Toluidine Blue O, C15H16N3SCl305.8 [92-31-9]Use
ing the directions given in the test for Related compounds
a suitable grade.
under Propoxyphene Hydrochloride, inject a suitable volume
n-Triacontane, C30H62422.81 [638-68-6]Use a suit-
of the solution into the chromatograph, and record the
able grade.
chromatogram. Concomitantly record the chromatogram
2,4,6-Triamino-5-nitrosopyrimidine, C4H6N6O154.13
from the Standard preparation prepared as directed in the
Pink powder.
test for Related compounds: only one main peak is obtained
AssayDissolve about 34 mg, accurately weighed, in from the n-tricosane solution, and no minor peaks are ob-
50 mL of glacial acetic acid. Titrate with 0.1 N perchloric served at, or near, the peak positions obtained for propoxy-
acid VS, determining the endpoint potentiometrically. Per- phene, acetoxy, or carbinol in the chromatogram from the
form a blank determination, and make any necessary correc- Standard preparation.
tions. Each mL of 0.1 N perchloric acid is equivalent to TriethanolamineUse Trolamine (NF monograph).
15.41 mg of C4H6N6O. Not less than 97% is found. Triethylamine, (C2H5)3N101.19 [121-44-8]Color-
Tributyl Phosphate (Tri-n-butyl Phosphate), (C4H9)3PO4 less liquid. Slightly soluble in water. Miscible with alcohol,
266.31 [126-73-8]Clear, almost colorless liquid. Slightly with ether, and with cold water. Store in well-closed con-
soluble in water. Miscible with common organic solvents. tainers. Use a suitable grade with a content of not less than
Specific gravity: about 0.976. 99.5%.
Triethylamine Hydrochloride, C6H15N HCl137.65
Refractive index 831: between 1.4205 and 1.4225.
Tributylethylammonium Hydroxide, C14H33NO
231.42Use a suitable grade. AssayTransfer about 35 mg, accurately weighed, to a
Tributyrin (Glyceryl Tributyrate), C15H26O6302.36 suitable beaker, add 50 mL of glacial acetic acid, and dis-
[60-01-5]Colorless, oily liquid. Insoluble in water; very sol- solve by stirring. Add 5 mL of mercuric acetate TS, with stir-
uble in alcohol and in ether. ring. When solution is complete, titrate with 0.1 N perchlo-
ric acid VS, determining the endpoint potentiometrically.
Perform a blank titration, and make any necessary correc-
a flame-ionization detector, nitrogen being used as the car-
13.77 mg of C6H15N HCl. Not less than 97.5% of C6H15N
HCl is found.
a 3-mm 1.8-m stainless steel column containing phase G4
on support S1A; the injection port temperature is main- Melting point 741: between 256 and 259, with de-
tained at 270; and the detector temperature is maintained composition.
at 300. The area of the tributyrin peak is not less than 98% Triethylamine Phosphate (Triethylammonium Phosphate),
of the total peak area. C6H15N H3O4P199.19Use a suitable grade.
[NOTEA suitable grade is available from www.
tciamerica.com, catalog number T1300.]
20.
Triethylene Glycol, C6H14O4150.17 [112-27-6]Col-
Acid contentTransfer 1.0 g, accurately weighed, to a
orless to pale yellow liquid. Is hygroscopic. Miscible with
beaker, add 75 mL of methanol, and dissolve by stirring.
water, with alcohol, and with toluene.
When dissolution is complete, add 25 mL of water, and ti-
AssayInject an appropriate test specimen into a suitable
trate with 0.05 N potassium hydroxide VS, using phenol-
gas chromatograph equipped with a flame-ionization detec-
phthalein TS as the indicator. Perform a blank determina-
tor (see Chromatography 621), helium being used as the
tion, and make any necessary correction. Each mL of 0.05 N
potassium hydroxide is equivalent to 88.1 mg of butyric
ble: a 3-mm 1.85-m stainless steel column packed with
acid: not more than 0.5% is found.
support S2; the injection port, column, and detector tem-
Trichloroacetic Acid, CCl3COOH163.39 [76-03-9]
peratures are maintained at 250, 230, and 310, respec-
tively. The area of the C6H14O4 peak is not less than 97% of
TrichloroethaneSee Methyl Chloroform.
Trichlorofluoromethane, CCl3F137.37 [75-69-4]
Colorless liquid. Refractive index 831: between 1.4550 and 1.4570, at
20.
Triethylenediamine (1,4-Diazobicyclo[2.2.2]octane),
C6H12N2112.17 [280-57-9]Use a suitable grade with a
thermal conductivity detector, helium being used as the car-
Trifluoroacetic Acid, C2HF3O2114.02 [76-05-1]
a 2.0-mm 1.8-m glass column packed with 10% G1 phase
Colorless liquid. Miscible with ether, with acetone, with eth-
on support S1A; the injection port temperature is main-
anol, with benzene, with carbon tetrachloride, and with
hexane.
programmed to rise 3 per minute to 50. The area of the AssayDissolve about 300 mg, accurately weighed, in
CCl3F peak is not less than 99% of the total peak area. 25 mL of water and 25 mL of alcohol. Titrate with 0.1 N
sodium hydroxide VS, determining the endpoint potentio-
metrically. Perform a blank determination, and make any
20.
necessary corrections. Each mL of 0.1 N sodium hydroxide
TrichlorotrifluoroethaneUse a suitable grade.
is equivalent to 11.40 mg of C2HF3O2. Not less than 99% is
[NOTEA suitable preparation, listed as Freon-TF aerosol,
found.
is available from E. I. du Pont de Nemours and Co., Wil-
Trifluoroacetic Anhydride, (F3CCO)2O210.03
mington, DE 19898.]
[407-25-0]Colorless liquid. Boils between 40 and 42. Ex-
n-Tricosane, C23H48324.63 [638-67-5]Colorless or
tremely volatile. Avoid exposure to air or water.
white, more or less translucent mass, showing a crystalline
structure. Has a slightly greasy feel. Insoluble in water and AssayTransfer about 0.8 g, accurately weighed, to a
in alcohol; soluble in chloroform, in ether, in volatile oils, glass-stoppered flask containing 50 mL of methanol. Add
and in most warm fixed oils; slightly soluble in dehydrated 500 mg of phenolphthalein, and titrate with 0.1 N sodium
alcohol. Boils at about 380.
methoxide VS to a pink endpoint. Calculate A by the Residue on ignition (Reagent test)Ignite 1 g with 0.5 mL
formula: of sulfuric acid: the residue weighs not more than 10 mg
(1%).
V/W TrimethylchlorosilaneSee Chlorotrimethylsilane.
2,2,4-Trimethylpentane (Isooctane), C8H18114.23
in which V is the volume, in mL, of 0.1 N sodium methox- [540-84-1]Use ACS reagent grade.
ide and W is the weight, in mg, of test specimen. To a 2,4,6-Trimethylpyridine (5-Collidine), C8H11N121.18
second glass-stoppered flask containing 50 mL of a mixture [108-75-8]Clear, colorless liquid. Soluble in cold water
of dimethylformamide and water (1:1) transfer 0.4 g, accu- and less soluble in hot water; soluble in alcohol, in chloro-
rately weighed, of the specimen under test, add 500 mg of form, and in methanol. Miscible with ether.
phenolphthalein, and titrate with 0.1 N sodium hydroxide
AssayInject an appropriate test specimen into a suitable
VS to a pink endpoint. Calculate B by the formula:
gas chromatograph (see Chromatography 621), helium be-
ing used as a carrier gas. The following conditions have
V
1
/W
1
been found suitable: a 3-mm 1.85-m stainless steel col-
umn containing phase G16 on support S1A; the injection
in which V
1
is the volume, in mL, of 0.1 N sodium hydrox-
port, column, and detector temperatures are maintained at
ide and W
1
is the weight, in mg, of test specimen. Calculate
180, 165, and 270, respectively; and a flame-ionization
the percentage of (F3CCO)2O by the formula:
detector is used. The area of the C8H11N peak is not less
2100.3(B A)
20.
Not less than 97% is found. If 2A is greater than B, calculate
N-(Trimethylsilyl)-imidazole, C6H12N2Si140.26
the percentage of F3CCOOH by the formula:
[18156-74-6]A clear, colorless, to light yellow liquid.
1140.3(2A B) Refractive index 831: between 1.4744 and 1.4764 at
20.
,,-Trifluoro-p-cresol (4-hydroxybenzotrifluoride, Trimethyltin Bromide, C3H9BrSn243.72
4-trifluoromethylphenol), C7H5F3O162.11 [402-45-9] [1066-44-0]Use a suitable grade.
2,4,6-Trinitrobenzenesulfonic Acid, C6H2(NO2)3SO3H Melting range 741: between 48 and 52.
3H2O347.21 [2508-19-2]Pale yellow to tan crystals. 2,2,2-Trifluoroethanol, CF3CH2OH100.04
Use a suitable grade. Also available as a 5% (w/v) or a 1 M [75-89-8]Colorless liquid.
aqueous solution.
TrinitrophenolSee Picric Acid.
Trioctylphosphine Oxide, C24H51PO386.63
[78-50-2]White, crystalline powder. Insoluble in water; sol-
uble in organic solvents.
Melting range 741: between 54 and 56. of phase G2; the injection port temperature is maintained at
1,3,5-Triphenylbenzene, (C6H5)3C6H3306.41 100; the detector temperature is maintained at 150; and
[612-71-5]White to off-white powder. the column temperature is maintained at 0 and pro-
grammed to rise 10 per minute to 150. The area of the Melting range 741: between 172 and 175.
CF3CH2OH peak is not less than 99% of the total peak area. Triphenylmethane, C19H16244.34 [519-73-3]Light
brown powder. Boiling range: between 77 and 80.
2,2,2-Trifluoroethyldifluoromethyl Ether (Difluoromethyl- AssayInject an appropriate volume into a gas chromat-
2,2,2-trifluoroethyl ether), C3H3F5O150.05Clear liquid. ograph (see Chromatography 621) equipped with a flame-
Use a suitable grade. ionization detector, helium being used as the carrier gas.
The following conditions have been found suitable: a 0.25- Boiling range: between 28 and 30.
mm 30-m capillary column coated with a 1-m layer of [NOTEA suitable grade is available from PCR Incorpo-
phase G2; the injection port temperature is maintained at rated, P.O. Box 1466, Gainesville, FL 32602. Tel:
300; the detector temperature is maintained at 300; and 904-376-8246. The catalogue number is 17151-2.]
the column temperature is maintained at 200 and pro- (m-Trifluoromethylphenyl) Trimethylammonium
grammed to rise 10 per minute to 300. The area of the Hydroxide in MethanolUse a suitable grade.
C19H16 peak is not less than 99% of the total peak area. [NOTEA suitable grade is available as Meth-Prep II
from Alltech, www.alltechweb.com.] Melting range 741: between 92 and 94.
5-(Trifluoromethyl)uracil, C5H3F3N2O2180.08 Triphenylmethanol, C19H16O260.34 [76-84-6]
[54-20-6]White to off-white powder. White to off-white powder.
AssayWhen tested by thin-layer chromatography, with AssayInject an appropriate volume into a gas chromat-
the use of plates coated with chromatographic silica gel ograph (see Chromatography 621) equipped with a flame-
mixture, a developing system consisting of chloroform, ionization detector, helium being used as the carrier gas.
methanol, and acetic acid (17:2:1), and examined visually The following conditions have been found suitable: a 0.25-
and under long-wavelength UV light, a single spot is exhib- mm 30-m capillary column coated with a 1-m layer of
ited. phase G2; the injection port temperature is maintained at
Trifluorovinyl Chloride Polymer (Fluorolube; 1-Chloro-1,2, 280; the detector temperature is maintained at 300; and
2-trifluoro-ethene Homopolymer), (C2ClF3)xUse a suitable the column temperature is maintained at 180. The area of
grade. the C19H16O peak is not less than 96.5% of the total peak
[NOTEA suitable grade is available as catalog number area.
Z123552 from www.sigma-aldrich.com.] Triphenyltetrazolium Chloride, C19H15ClN4334.80
Trimethylacethydrazide Ammonium Chloride (Betaine [298-96-4]White to yellowish, crystalline powder. Soluble
Hydrazide Chloride; Girard Reagent T), in about 10 parts of water and of alcohol; slightly soluble in
[(CH3)3N
+
CH2CONHNH2]Cl
167.64 [123-46-6]Color- acetone; insoluble in ether. Usually contains solvent of crys-

less or white crystals. Freely soluble in water. One g dis- tallization, and when dried at 105 it melts at about 240,
solves in about 25 mL of alcohol. Insoluble in chloroform with decomposition.
and in ether. Hygroscopic.
SolubilitySeparate 100-mg portions dissolve completely
Melting range 741: between 185 and 192, deter- in 10 mL of water and in 10 mL of alcohol, respectively, to
mined after recrystallization from hot alcohol, if necessary. yield solutions that are clear, or practically so.
Loss on drying 731Dry it at 105 to constant weight: fate TS, and add to the combined filtrate 0.5 mL of sodium
it loses not more than 5.0% of its weight. nitrite solution (1 in 20): no red color is produced within
15 minutes.
Tuberculin, Purified Protein Derivative (Tuberculin
100 mg.
PPD)Derived from the human strain of Mycobacterium tu-
SensitivenessDissolve 10 mg in 10 mL of dehydrated al-
berculosis, and available either as a solution or as a lyophi-
cohol (A). Then dissolve 10 mg of dextrose in 20 mL of de-
lized powder. For lyophilized powder, reconstitute according
hydrated alcohol (B). To 0.2 mL of B add 1 mL of dehy-
to the manufacturers instructions using the diluent pro-
drated alcohol and 0.5 mL of dilute tetramethylammonium
vided by the manufacturer. Solutions may contain a stabi-
hydroxide TS (1 volume diluted with 9 volumes of dehy-
lizer and a preservative. One Tuberculin Unit (TU) is equiva-
drated alcohol), then add 0.2 mL of A: a pronounced red
lent to 0.02 g of Tuberculin PPD.
color develops within about 10 minutes.
Tubocurarine Chloride (7,12-Dihydroxy-6,6-dimethoxy-
Tris(2-aminoethyl)amine, C6H18N4146.23
2,2,2-trimethyltubocuraranium Chloride), C37H42Cl2N2O6
[4097-89-6]Yellow liquid. Soluble in methanol.
681.65 [6989-98-6]Use a suitable grade with an assay
AssayDissolve about 80 mg in 30 mL of methanol. Add
result between 98.0% and 102.0%.
40 mL of water, and titrate with 1 N hydrochloric acid, de-
[NOTEA suitable grade is available from Acros Organics,
termining the endpoint potentiometrically. Perform a blank
catalog number 24349 at www.acros.com.]
determination, and make any necessary correction. Each mL
Tungstic Acid, H2WO4249.85 [7783-03-1]Use a
of 1 N hydrochloric acid is equivalent to 48.75 mg of
C6H18N4. Not less than 98.0% is found.
L-Tyrosine Disodium, C9H9NO3Na2225.2
Refractive index 831: between 1.4956 and 1.4986 at [69849-45-6]Off-white to tan powder. Use a suitable
20. grade.
Tris(hydroxymethyl)aminomethane [77-86-1]Use ACS Uracil, C4H4N2O2112.09 [66-22-8]White to cream-
reagent gradeSee also Tromethamine. colored, crystalline powder. Melts above 300. One g dis-
Tris(hydroxymethyl)aminomethane Acetate, C4H11O3 solves in about 500 mL of water; less soluble in alcohol; sol-
NCH3181.19 [6850-28-8]White powder with lumps. uble in ammonia TS and in sodium hydroxide TS. Its solu-
Use a suitable grade. tions yield no precipitate with the usual alkaloidal
Tris(hydroxymethyl)aminomethane Hydrochloride, precipitants.
C4H11NO3 HCl157.60 [1185-53-1]Colorless crystals.
100 mg.
N-Tris(hydroxymethyl)methylglycine, C6H13NO5179.2
[5704-04-1]White, crystalline powder. Use a suitable
grade.
Uranyl Acetate (Uranium Acetate), UO2(C2H3O2)2 2H2O
Tritirachium Album Proteinase KUse a suitable grade.
[NOTEA suitable grade can be obtained from Qiagen,
Urea, NH2CONH260.06 [57-13-6]Use ACS reagent
Inc., www.qiagen.com.]
grade.
Tromethamine [Tris(hydroxymethyl)aminomethane; THAM;
Urethane (Ethyl carbamate), C3H7NO289.09
2-Amino-2-(hydroxymethyl)-1,3-propanediol], C4H11NO3
[51-79-6]White powder with chunks. Freely soluble in
121.14Use ACS reagent grade Tris(hydroxymethyl)amino-
water.
methane.
Tropaeolin OO (Acid Orange 5), C18H14N3NaO3S375.38
Uridine, C9H12N2O6244.20 [58-96-8]White pow-
[554-73-4]Orange-yellow scales, or yellow powder. Solu-
der.
ble in water.
Assay
pH range: from 1.4 (red) to 2.6 (yellow).
Tropic Acid, C9H10O3166.18 [529-64-6]Use a suit- MOBILE PHASEPrepare a mixture of methanol and 0.2 M
able grade. ammonium acetate (10:90), and adjust with phosphoric
Tropine, C8H15NO141.2 [120-29-6]Use a suitable acid to a pH of 7.0.
grade.
TEST SOLUTION: 0.5 mg per mL in water.
Trypan Blue (Direct Blue 14), C34H24N6Na4O14S4960.8
PROCEDUREInject about 20 L of the Test solution into a
liquid chromatograph (see Chromatography 621), equipped
TryptoneUse Pancreatic Digest of Casein.
with a 280-nm detector and a 4.6-mm 15-cm column
L-Tryptophane, C11H12N2O2204.23 [73-22-3]White
that contains packing L1. The flow rate is about 2.0 mL per
or not more than slightly yellow leaflets or powder. One g
minute. The area of the C9H12N2O6 peak is not less than
dissolves in about 100 mL of water; soluble in dilute acids
99% of the total peak area.
and in solutions of the alkali hydroxides; slightly soluble in
alcohol.
Valeric Acid, C5H10O2102.13 [109-52-4]Clear, col-
AssayAccurately weigh about 300 mg, dissolve in a
orless liquid.
mixture of 3 mL of formic acid and 50 mL of glacial acetic
AssayAccurately weigh about 500 mg, transfer to a
acid, add 2 drops of crystal violet TS, and titrate with 0.1 N
suitable container, add 30 mL of water, and mix. Add 40 mL
perchloric acid VS to a green endpoint. Each mL of 0.1 N
of water, and mix. Add phenolphthalein TS, and titrate with
perchloric acid is equivalent to 20.42 mg of C11H12N2O2. Be-
tween 98.0% and 102.0%, calculated on the dried basis, is
found.
Specific rotation 781: between 30.0 and 33.0, deter-
Valerophenone, C11H14O162.23 [1009-14-9]Col-
mined in a solution containing 1.0 g of test specimen, previ-
orless liquid.
ously dried at 105 for 3 hours, in 100 mL.
0.1%.
TyrosineDissolve 100 mg in 3 mL of diluted sulfuric injection port temperature is maintained at 250; the detec-
acid, add 10 mL of mercuric sulfate TS, and heat on a steam tor temperature is maintained at 300; and the column tem-
bath for 10 minutes. Filter, wash with 5 mL of mercuric sul- perature is maintained at 150 and programmed to rise 10
per minute to 300. The area of the C11H14O peak is not less CH3COOCH=CH2 peak is not less than 99% of the total
than 98% of the total peak area. peak area.
2-Vinylpyridine, C7H7N105.14 [100-69-6].
Boiling range: between 79 and 82, at 29 mm of mer-
Boiling range: between 105 and 107, at a pressure of
cury.
5 mm of mercury.
Vanadium Pentoxide, V2O5181.88 [1314-62-1] Density: 0.975 at 25.
Fine, yellow to orange-yellow powder. Slightly soluble in
water; soluble in concentrated acids and in alkalies; insolu-
Vinylpyrrolidinone (1-Vinyl-2-pyrrolidinone; 1-Vinyl-
ble in alcohol.
2-pyrrolidone; N-Vinylpyrrolidinone; N-Vinylpyrrolidone),
AssayTransfer about 400 mg, accurately weighed, to a C6H9NO111.14 [88-12-4]Colorless liquid.
500-mL conical flask, and add 150 mL of water and 30 mL
of dilute sulfuric acid (1 in 2). Boil the solution on a hot
plate for 5 minutes, add 50 mL of water, and continue boil-
ing until a yellow solution is obtained. Transfer the hot plate
and the flask to a well-ventilated hood, and bubble sulfur
dioxide gas through the solution for 10 minutes, or until the
G2; the injection port temperature is maintained at 250;
solution is a clear, brilliant blue color. Rinse the gas delivery
the detector temperature is maintained at 300; and the
tube into the flask with a few mL of water, then bubble
carbon dioxide gas through the solution for 30 minutes
while continuing to boil the solution gently. Cool the solu-
C6H9NO peak is not less than 99.0% of the total peak area.
tion to about 80, and titrate with 0.1 N potassium perman-
Water, Method 1 921: not more than 0.1%, determined
ganate VS to a yellow-orange endpoint. Perform a complete
on 2.5 g, using a mixture of 50 mL of methanol and 10 mL
of butyrolactone as the solvent.
Each mL of 0.1 N potassium permanganate is equivalent to
[NOTEA suitable grade is available from Merck KGaA/
9.095 mg of V2O5. Not less than 99.5% is found.
EMD chemicals, catalogue number 8.08518.0250, www.
Vanadyl Sulfate, VOSO4 xH2O (anhydrous)163.00
emdchemicals.com.]
[27774-13-6]Blue, hygroscopic crystals. Slowly and usually
Washed SandSee Sand, Washed.
incompletely soluble in water.
Water, Ammonia-Free, H2O18.02Use High-Purity
AssayAccurately weigh about 400 mg of the dried test
Water as defined for Chemical Resistance under Containers
specimen obtained in the test for Water, and transfer with
Glass 660.
15 to 20 mL of water into a beaker. Add 3 mL of sulfuric
Water, Carbon Dioxide-FreeSee Water, in the introduc-
acid, cover the beaker with a watch glass, and heat on a
tory section.
steam bath until all dissolves. Cool, dilute with 125 mL of
Water, DeaeratedSee Water, in the introductory sec-
water, and titrate with 0.1 N potassium permanganate VS
tion.
to the production of a pinkish color that persists for 1 min-
Water Vapor Detector TubeA fuse-sealed glass tube so
ute: each mL of 0.1 N potassium permanganate is equiva-
designed that gas may be passed through it and containing
lent to 16.30 mg of VOSO4. Not less than 97% is found.
WaterDry about 1 g, accurately weighed, at 220 to tor, which consists of a selenium sol in suspension in sulfuric
constant weight: it loses not more than 50.0% of its weight. acid.
Pentavalent vanadiumHeat 1 g, accurately weighed, Measuring range: 5 to 250 mg per cubic meter.
with 50 mL of water and 5 mL of hydrochloric acid in a flask [NOTEAvailable from Draeger Safety, Inc., www.draeger.
until dissolved. Cool, add 2 g of potassium iodide, insert the com, or from Gastec Corp., www.gastec.co.jp, distributed in
stopper, and allow to stand for 30 minutes. Add 50 mL of the USA by www.nextteq.com.]
water, and titrate the liberated iodine with 0.1 N sodium Wrights Stain [68988-92-1]A mixture of methylene
thiosulfate VS, adding 3 mL of starch TS as the indicator. blue, methylene azure, and the eosinates of both, available
Correct for the volume of thiosulfate consumed by a blank. as a solid and as a solution in methanol. Use a suitable
Each mL of 0.1 N thiosulfate is equivalent to 5.095 mg of grade. [NOTEIf a solid is used, dissolve 6.0 g of Wrights
vanadium (V). Not more than 0.5% is found, calculated on stain powder (CAS# 68988-92-1, dark green powder) and
the dried basis. 0.6 g of Giemsa stain powder (CAS# 51811-82-6, dark
green to black powder or crystals) in 1000 mL of methanol.
Substances not precipitated by ammoniaDissolve 1.0 g
Stir overnight, and filter before use.]
by heating with 20 mL of water and 2 mL of hydrochloric
Xanthine, C5H4N4O2152.11 [69-89-6]White, crys-
acid. Dilute with water to about 75 mL, and neutralize to
talline powder. Decomposes on heating. Slightly soluble in
litmus paper with ammonia TS. Transfer the solution to a
water and in alcohol; soluble in sodium hydroxide TS; spar-
cylinder, slowly add 5 mL of ammonia TS and sufficient
ingly soluble in diluted hydrochloric acid. When subjected
water to make 100 mL, and allow to stand overnight. De-
to the murexide reaction, a purple color is produced with
cant 50 mL of the supernatant through a filter, add 5 drops
the ammonia, but on the subsequent addition of fixed alkali
of sulfuric acid, evaporate to dryness, and ignite: the residue
hydroxides, the color is not discharged but is changed to
violet.
Vinyl Acetate, CH3COOCH=CH286.09 [108-05-4]
Liquid. Residue on ignition (Reagent test): negligible, from
100 mg.
ograph (see Chromatography 621) equipped with a flame- Loss on drying 731Dry it at 105 for 2 hours: it loses
ionization detector, helium being used as the carrier gas. not more than 1% of its weight.
The following conditions have been found suitable: a 0.25- Xanthydrol, C13H10O2198.22 [90-46-0]Pale yellow,
mm 30-m capillary column coated with a 1-m layer of crystalline powder. Insoluble in water; soluble in alcohol, in
G2; the injection port temperature is maintained at 100; chloroform, and in ether. Soluble in glacial acetic acid,
the detector temperature is maintained at 300; and the forming a practically colorless solution; but when the pow-
column temperature is maintained at 100 and pro- der is treated with diluted hydrochloric acid, a lemon-yellow
grammed to rise 10 per minute to 150. The area of the color is produced.
Melting range 741: between 121 and 123. tions, clarified, and dried to a reddish-yellow to brown pow-
der, having a characteristic but not putrescent odor. Soluble
Residue on ignitionIgnite 500 mg with 0.5 mL of sulfuric
in water, forming a yellow to brown solution, having a
acid: the residue weighs not more than 10 mg (2.0%).
slightly acid reaction. Contains no added carbohydrate. One
Xylene, C8H10106.17 [1330-20-7]Use ACS reagent
g represents not less than 7.5 g of yeast.
grade.
m-Xylene, C6H4(CH3)2106.17 [108-38-3]Clear, col- Loss on drying 731Dry it at 105 to constant weight:
orless, flammable liquid. Insoluble in water; miscible with it loses not more than 6% of its weight.
alcohol and with ether. Use a suitable grade.
Residue on ignitionIgnite 500 mg with 1 mL of sulfuric
o-Xylene, C8H10106.17 [95-47-6]Clear, colorless,
acid: the residue weighs not more than 75 mg (15%).
mobile, flammable liquid. Insoluble in water; miscible with
alcohol and with ether.
boiling: no precipitate is formed.
Chloride (Reagent test)It shows not more than 5% of
Cl, calculated as sodium chloride.
tions have been found suitable for assaying the substance: A
3-mm 1.8-m stainless steel column packed with 1.75%
hydrated aluminum silicate plus 5.0% diisodecyl phthalate
105 to constant weight: between 7.2% and 13.0% of N is
on support S1. Helium, having a flow rate of about 27.5 mL
found.
per minute is the carrier gas, the detector temperature is
4
cfu/g about 280, the injection port temperature is about 180,
Yellow Mercuric OxideSee Mercuric Oxide, Yellow. and the column temperature is 80. A flame-ionization de-
Zinc, ZnAt. Wt. 65.39 [7440-66-6]Use ACS rea- tector is employed.
gent grade.
Refractive index 831: between 1.5040 and 1.5060,
Zinc, ActivatedPlace zinc pellets in a conical flask, and
at 20.
add a sufficient quantity of a 50-ppm solution of chloropla-
p-Xylene, C8H10106.17 [106-42-3]Colorless liquid.
tinic acid to cover all pellets. Allow the metal to remain in
contact with the solution for 10 min, drain, wash, and dry
immediately.
AssayTo 5 g of activated zinc add 15 mL of hydrochlo-
ric acid, 25 mL of water, 0.1 mL of stannous chloride solu-
tion (235 mg/mL in hydrochloric acid), and 5 mL of potas-
phase G14; the column temperature is maintained at 50
sium iodide solution (166 mg/mL in water). No stain is
and programmed to rise 10 per minute to 100; the injec-
produced when in contact with mercuric bromide paper.
tion port is maintained at 130; and the detector is main-
Repeat the test for arsenic, using the same reagents and
tained at 300. The area of the C8H10 peak is not less than
adding a solution containing 1 g of arsenic. An appreciable
99% of the total peak area.
stain appears when in contact with mercuric bromide paper.
Zinc Acetate, Zn(CH3COO)2 2H2O219.51
Xylene Cyanole FF, C25H27N2NaO6S2538.61
[2650-17-1]Gray-blue to dark blue powder. Soluble in
Zinc AmalgamAdd 54 g of mossy or granular zinc to
water.
100 mL of mercury in a beaker. Heat, with stirring, on a hot
plate under a hood [Cautionmercury vapor is extremely
100-mL volumetric flask, dissolve in water, dilute with water
toxic] until solution of the zinc is complete or practically so.
to volume, and mix. Transfer 2.0 mL of the solution to a
Allow to cool to room temperature, and if necessary add
50-mL volumetric flask, dilute with pH 7.0 phosphate buffer
sufficient mercury to prevent solidification of the amalgam.
to volume (See Solutions, in this section), and mix. Using a
Transfer the amalgam to a glass-stoppered bottle, and shake
suitable spectrophotometer, 1-cm cells, and water as the
a few times with dilute hydrochloric acid (1 in 2), to remove
blank, record the absorbance of the solution at the wave-
any zinc oxide formed.
length of maximum absorbance at about 614 nm. From the
Zinc Chloride, Anhydrous, PowderedUse Zinc Chloride
observed absorbance, calculate the absorptivity (see Spectro-
(USP monograph) that has been dried and powdered.
photometry and Light-scattering 851): the absorptivity is not
Zinc Sulfate Heptahydrate, ZnSO4 7H2O287.54
less than 55.9, corresponding to about 83% of
[7446-20-0]Odorless crystals, granules, or powder.
C25H27N2NaO6S2.
Melting point 741: 100.
Density: 1.97.
Zirconyl Chloride, Octahydrate, BasicUse Zirconium
Xylose, C5H10O5150.13 [58-86-6]Use a suitable
Oxychloride.
grade.
Zirconyl Nitrate, ZrO(NO3)2231.23 [13826-66-9]
Yeast ExtractA water-soluble, peptone-like derivative of
yeast cells (Saccharomyces) prepared under optimum condi-
Indicators and Indicator Test Papers
containing an acidic group, the acid must first be neutral-
INDICATORS
ized with sodium hydroxide as follows:
Triturate 100 mg of the indicator in a smooth-surfaced
Indicators are required in the Pharmacopeial tests and as-
mortar with the volume of 0.05 N sodium hydroxide speci-
says either to indicate the completion of a chemical reaction
fied in the directions for preparing its Test Solution, or with
in volumetric analysis or to indicate the hydrogen-ion con-
the equivalent of 0.02 N sodium hydroxide. When the indi-
centration (pH) of solutions. The necessary solutions of in-
cator has dissolved, dilute the solution with carbon dioxide-
dicators are listed among the Test Solutions, abbreviated
free water to 200 mL (0.05%). Store the solutions in suitably
TS.
resistant containers, protected from light.
Solutions of indicators of the basic type and of the phtha-
leins are prepared by dissolving in alcohol. With indicators
USP 36 Indicators and Indicator Test Papers / Indicators 1207
Listed in ascending order of the lower limit of their range, Eosin Y (Indicator grade Eosin Y, Sodium te-
useful pH indicators are: thymol blue, pH 1.22.8; methyl trabromofluorescein), C20H6Br4Na2O5691.86 [17372-87-1]
yellow, pH 2.94.0; bromophenol blue, pH 3.04.6; bromo- Red to brownish-red pieces or powder. One g dissolves in
cresol green, pH 4.05.4; methyl red, pH 4.26.2; bromo- about 2 mL of water and in 50 mL of alcohol. Dye content
cresol purple, pH 5.26.8; bromothymol blue, pH 6.07.6; about 80%.
phenol red, pH 6.88.2; thymol blue, pH 8.09.2; and thy- Eriochrome Black T [Sodium 1-(1-Hydroxy-2-naphthy-
molphthalein, pH 8.610.0. lazo)5-nitro-2-naphthol-4-sulfonate], C20H12N3NaO7S
Alphazurine 2GUse a suitable grade. 461.38Brownish-black powder having a faint, metallic
Azo Violet [4-(p-Nitrophenylazo)resorcinol], C12H9N3O4 sheen. Soluble in alcohol, in methanol, and in hot water.
259.22Red powder. It melts at about 193, with decom-
SensitivenessTo 10 mL of a 1 in 200,000 solution in a
position.
mixture of equal parts of methanol and water add sodium
Bismuth SulfiteUse a suitable grade.
hydroxide solution (1 in 100) until the pH is 10: the solu-
Brilliant GreenSee Brilliant Green in the section Re-
tion is pure blue in color and free from cloudiness. Add
agents.
0.01 mg of magnesium ion (Mg): the color of the solution
Brilliant Yellow (C.I. 24890), C26H18N4Na2O8S592.49
changes to red-violet, and with the continued addition of
Orange to rust-colored powder. Soluble in water.
magnesium ion it becomes wine-red.
Loss on drying 731Dry it in vacuum at 60 for 1 hour: Eriochrome Black T TriturationGrind 200 mg of eri-
it loses not more than 5% of its weight. ochrome black T to a fine powder with 20 g of potassium
Bromocresol BlueUse Bromocresol Green. chloride.
Bromocresol Green (Bromocresol Blue; Tetrabromo-m-cre- LitmusBlue powder, cubes, or pieces. Partly soluble in
sol-sulfonphthalein), C21H14Br4O5S698.01White or pale water and in alcohol. Transition interval: from approximately
buff-colored powder. Slightly soluble in water; soluble in al- pH 4.5 to 8. Color change: from red to blue. Litmus is un-
cohol and in solutions of alkali hydroxides. Transition inter- suitable for determining the pH of alkaloids, carbonates, and
val: from pH 4.0 to 5.4. Color change: from yellow to blue. bicarbonates.
Bromocresol Green Sodium SaltUse a suitable grade. Malachite Green Oxalate, [C23H25N2
+
]2 [C2HO4
]2
Bromocresol Purple (Dibromo-o-cresolsulfonphthalein), C2H2O4927.00The oxalate salt, crystallized with oxalic
C21H16Br2O5S540.22White to pink, crystalline powder. acid, of a triphenylmethane dye. Dark-green powder, having
Insoluble in water; soluble in alcohol and in solutions of a metallic luster. Sparingly soluble in water; soluble in glacial
alkali hydroxides. Transition interval: from pH 5.2 to 6.8. acetic acid. Transition interval: from pH 0.0 to 2.0. Color
Color change: from yellow to purple. change: from yellow to green.
Bromocresol Purple Sodium Salt, C21H15Br2O5SNa Methyl Orange (Helianthin or Tropaeolin D),
562.20Black powder. Soluble in water. Transition interval: C14H14N3NaO3S327.33The sodium salt of dimethylami-
from pH 5.0 to 6.8. Color change: from greenish yellow to noazobenzene sulfonic acid or dimethylaminoazobenzene
purple-violet. sodium sulfonate. An orange-yellow powder or crystalline
scales. Slightly soluble in cold water; readily soluble in hot
water; insoluble in alcohol. Transition interval: from pH 3.2
Bromophenol Blue (3,3,5,5-Tetrabromophenol-
to 4.4. Color change: from pink to yellow.
sulfonphthalein), C19H10Br4O5S669.96Pinkish crystals. In-
Methyl Red (2-[[4-(Dimethylamino)phenyl]azo]benzoic Acid
soluble in water; soluble in alcohol and in solutions of alkali
Hydrochloride), 2-[4-(CH3)2NC6H4N:N]C6H4COOH HCl
hydroxides. Transition interval: from pH 3.0 to 4.6. Color
305.76Dark-red powder or violet crystals. Sparingly solu-
change: from yellow to blue.
ble in water; soluble in alcohol. Transition interval: from pH
Bromophenol Blue SodiumThe sodium salt of 3,3,5,
4.2 to 6.2. Color change: from red to yellow.
5 (Tetrabromophenolsulfonphthalein), C19H9Br4O5SNa
Methyl Red SodiumThe sodium salt of 2-[[4-(dimeth-
646.36Pinkish crystals. Soluble in water and in alcohol.
ylamino)phenyl]azo]benzoic acid. 2-[4-(CH3)2NC6H4N:N]
Transition interval: from pH 3.0 to 4.6. Color change: from
C6H4COONa291.28Orange-brown powder. Freely solu-
yellow to blue.
ble in cold water and in alcohol. Transition interval: from pH
Bromothymol Blue (3,3-Dibromothymolsulfonphthalein),
4.2 to 6.2. Color change: from red to yellow.
C27H28Br2O5S624.38Cream-colored powder. Insoluble in
Methyl Yellow (p-Dimethylaminoazobenzene), C14H15N3
water; soluble in alcohol and in solutions of alkali hydrox-
225.29Yellow crystals, melting between 114 and 117.
ides. Transition interval: from pH 6.0 to 7.6. Color change:
Insoluble in water; soluble in alcohol, in benzene, in chloro-
from yellow to blue.
form, in ether, in dilute mineral acids, and in oils. Transition
Congo RedSee Congo Red in the section Reagents.
interval: from pH 2.9 to 4.0. Color change: from red to
Cresol Red (o-Cresolsulfonphthalein), C21H18O5S382.43
yellow.
Red-brown powder. Slightly soluble in water; soluble in al-
p-Naphtholbenzein (4-[-(4-Hydroxy-1-naph-
cohol and in dilute solutions of alkali hydroxides. Transition
thyl)benzylidene]-1(4H)-naphthalenone), (4-HOC10H6)
interval: from pH 7.2 to 8.8. Color change: from yellow to
C(:C10H6-4:O)(C6H5)374.43Reddish brown powder. In-
red.
soluble in water; soluble in alcohol, in benzene, in ether,
Crystal Violet (Hexamethyl-p-rosaniline Chloride),
and in glacial acetic acid. Transition interval: from pH 8.8 to
C25H30ClN3407.98Dark-green crystals. Slightly soluble in
10.0. Color change: from orange to green.
water; sparingly soluble in alcohol and in glacial acetic acid.
Neutral Red (3-Amino-7-dimethylamino-2-methylphenazine
Its solutions are deep violet in color.
Monohydrochloride), C15H16N4 HCl288.78Reddish to ol-
SensitivenessDissolve 100 mg in 100 mL of glacial acetic
ive-green, coarse powder. Sparingly soluble in water and in
acid, and mix. Pipet 1 mL of the solution into a 100-mL
alcohol. Transition interval: from pH 6.8 to 8.0. Color
volumetric flask, and dilute with glacial acetic acid to vol-
change: from red to orange.
ume: the solution is violet-blue in color and does not show
Nile Blue Hydrochloride (Nile Blue A, as the hydrochloride;
a reddish tint. Pipet 20 mL of the diluted solution into a
5-Amino-9- (diethylamino)benzo[a]phenoxazin-7-ium chloride),
beaker, and titrate with 0.1 N perchloric acid VS, adding the
C20H20ClN3O353.85Slightly soluble in alcohol and in
perchloric acid slowly from a microburet: not more than
glacial acetic acid. Transition interval: from pH 9.0 to 13.0.
0.10 mL of 0.1 N perchloric acid is required to produce an
Color change: from blue to pink.
emerald-green color.
Oracet Blue B (Solvent Blue 19)A mixture of 1-methyl-
4,5-Dihydroxy-3-(p-sulfophenylazo)-2,7-
amino-4-anilinoanthraquinone (C21H16N2O2) and 1-amino-
naphthalenedisulfonic Acid, Trisodium SaltSee 2-
4-anilinoanthraquinine (C20H14N2O2). Where used for titra-
(4-Sulfophenylazo)-1,8-dihydroxy-3,6-naphthalenedisulfonic
tion in non-aqueous media, it changes from blue (basic)
Acid, Trisodium Salt.
through purple (neutral) to pink (acidic).
1208 Indicators / Indicators and Indicator Test Papers USP 36
Phenol Red [4,4-(3H-2,1-Benzoxathiol-3-ylidene)diphenol, Phosphate (Reagent test)Cut 5 strips into small pieces,
S,S-Dioxide], C19H14O5S354.38Crystalline powder, vary- mix with 500 mg of magnesium nitrate in a porcelain cruci-
ing in color from bright to dark red. Very slightly soluble in ble, and ignite. To the residue add 5 mL of nitric acid, and
water; freely soluble in solutions of alkali carbonates and evaporate to dryness: the residue shows not more than
hydroxides; slightly soluble in alcohol. Transition interval: 0.02 mg of PO4.
from pH 6.8 to 8.2. Color change: from yellow to red. Residue on ignitionIgnite carefully 10 strips of the paper
Phenolphthalein [3,3-Bis(p-hydroxyphenyl)phthalide], to constant weight: the weight of the residue corresponds
C20H14O4318.32White or faintly yellowish-white, crystal- to not more than 0.4 mg per strip of about 3 square cm.
line powder. Insoluble in water; soluble in alcohol. Transition Rosin acidsImmerse a strip of the blue paper in a solu-
interval: from pH 8.0 to 10.0. Color change: from colorless tion of 100 mg of silver nitrate in 50 mL of water: the color
to red. of the paper does not change in 30 seconds.
Quinaldine Red (5-Dimethylamino-2-styrylethylquinolinium SensitivenessDrop a 10- to 12-mm strip into 100 mL of
Iodide), C21H23IN2430.33Dark blue-black powder. Spar- 0.0005 N acid contained in a beaker, and stir continuously:
ingly soluble in water; freely soluble in alcohol. Melts at the color of the paper is changed within 45 seconds. The
about 260, with decomposition. Transition interval: from 0.0005 N acid is prepared by diluting 1 mL of 0.1 N hydro-
pH 1.4 to 3.2. Color change: from colorless to red. chloric acid with freshly boiled and cooled purified water to
2-(4-Sulfophenylazo)-1,8-dihydroxy-3,6-naphtha- 200 mL.
lenedisulfonic Acid, Trisodium Salt (4,5-Dihydroxy-3-(p- Litmus Paper, RedUsually about 6 50 mm in size. Red
sulfophenylazo)-2,7-naphthalenedisulfonic Acid, Trisodium litmus paper meets the requirements of the tests for Phos-
Salt), C16H9N2O11S3Na3570.42Red powder. Soluble in phate, Residue on ignition, and Rosin acids, under Litmus Pa-
water. per, Blue.
Thymol Blue (Thymolsulfonphthalein), C27H30O5S SensitivenessDrop a 10- to 12-mm strip into 100 mL of
466.59Dark-colored, crystalline powder. Slightly soluble in 0.0005 N sodium hydroxide contained in a beaker, and stir
water; soluble in alcohol and in dilute alkali solutions. Acid continuously: the color of the paper changes within
Transition interval: from pH 1.2 to 2.8. Color change: from 30 seconds. The 0.0005 N sodium hydroxide is prepared by
red to yellow. AlkalineTransition interval: from pH 8.0 to diluting 1 mL of 0.1 N sodium hydroxide with freshly boiled
9.2. Color change: from yellow to blue. and cooled purified water to 200 mL.
Thymolphthalein, C28H30O4430.54White to slightly Mercuric Bromide Test PaperPlace a 50 mg/mL solu-
yellow, crystalline powder. Insoluble in water; soluble in al- tion of mercuric bromide in dehydrated alcohol in a dish,
cohol and in solutions of alkali hydroxides. Transition inter- and immerse in it pieces of white filter paper weighing
val: from pH 9.3 to 10.5. Color change: from colorless to 80 g/m
2
(speed of filtration = filtration time expressed in s
blue. for 100 mL of water at 20 with a filter surface of 10 cm
2
Xylenol Orange, (N,N-[3H-2,1-Benzoxathiol-3-ylidenebis- and a constant pressure of 6.7 kPa; 4060 s), each measur-
[(6-hydroxy-5-methyl-3,1-phenylene)methylene]]bis[N-(carboxy- ing 1.5 cm by 20 cm and folded in the middle. Allow the
methyl)glycine]S,S-dioxide), C31H28N2Na4O13S760.58Or- excess of liquid to drain, and allow the paper to dry, pro-
ange powder. Soluble in alcohol and in water. In acid solu- tected from light, suspended over a nonmetallic thread. Dis-
tion, it is lemon-yellow in color, and its metal complexes are card 1 cm from each end of each strip, and cut the remain-
intensely red. It yields a distinct endpoint where a metal der into 1.5-cm squares or discs of 1.5-cm diameter. Store
such as bismuth, cadmium, lanthanum, lead, mercury, scan- in a glass-stoppered container wrapped with black paper.
dium, thorium, or zinc is titrated with edetate disodium. Methyl GreenIodomercurate PaperImmerse thin
strips of suitable filter paper in a 40 g per L solution of
methyl green, and allow to air-dry. Immerse the strips for
INDICATORS AND TEST PAPERS
1 hour in a solution containing 140 g per L of potassium
iodide and 200 g per L of mercuric iodide. Wash with water
Indicator and test papers are strips of paper of suitable
until the washings are practically colorless, and allow to air-
dimension and grade (see Filter Paper, Quantitative, in the
dry. Store protected from light, and use within 48 hours.
section Reagents) impregnated with an indicator or a rea-
Methyl Yellow PaperUse a 1 in 2000 solution of
gent that is sufficiently stable to provide a convenient form
methyl yellow in alcohol.
of the impregnated substance. Some test papers may be
pH Indicator Paper, Short-RangeUse a suitable grade.
obtained from commercial sources of laboratory supplies.
Phenolphthalein PaperUse a 1 in 1000 solution of
Those required in Pharmacopeial tests and assays may be
phenolphthalein in diluted alcohol.
prepared as directed in the following paragraphs, by means
Starch Iodate PaperUse a mixture of equal volumes of
of the solutions specified, or to meet the tests set forth
starch TS and potassium iodate solution (1 in 20).
herein under the individual titles.
Starch Iodide PaperUse a solution of 500 mg of potas-
Treat strong, white filter paper with hydrochloric acid,
sium iodide in 100 mL of freshly prepared starch TS.
and wash with water until the last washing no longer shows
Thiazole Yellow PaperUse a 1 in 2000 solution of thia-
an acid reaction to methyl red. Then treat with ammonia
zole yellow in water.
TS, and wash again with water until the last washing is not
Turmeric PaperUse a solution prepared as follows:
alkaline to phenolphthalein.
Macerate 20 g of powdered turmeric, the dried root of Cur-
After thorough drying, saturate the paper with the proper
cuma longa Linn e (Fam. Zingiberaceae), with four 100-mL
strength of indicator solutions, and carefully dry in still air,
portions of cold water, decanting the clear liquid portion
unless otherwise specified, by suspending it from rods of
each time and discarding it. Dry the residue at a tempera-
glass or other inert material in a space free from acid, alkali,
ture not over 100. Macerate with 100 mL of alcohol for
and other fumes.
several days, and filter.
Cut the paper into strips of convenient size, and store the
SensitivenessDip a strip of the paper, of about 1.5-cm
papers in well-closed containers, protected from light and
length, in a solution of 1.0 mg of boric acid in 5 mL of
moisture.
water, previously mixed with 1 mL of hydrochloric acid. Af-
Cupric Sulfate Test PaperUse cupric sulfate TS.
ter 1 minute remove the paper from the liquid, and allow it
Lead Acetate Test PaperUsually about 6 80 mm in
to dry: the yellow color changes to brown. Then moisten
size. Use lead acetate TS, and dry the paper at 100, avoid-
the paper with ammonia TS: the color of the paper changes
ing contact with metal.
to greenish black.
Litmus Paper, BlueUsually about 6 50 mm in size. It
meets the requirements of the following tests.
USP 36 Solutions / Buffer Solutions 1209
Solutions
tablets, and buffer solids may be obtained from commercial
BUFFER SOLUTIONS
sources in convenient prepackaged form. Such preparations
are available for the entire working range in pharmaceutical
The successful completion of many Pharmacopeial tests
analysis, but are not recommended for pH meter standardi-
and assays requires adjustment to or maintenance of a spec-
zation (see pH 791).
ified pH by the addition of buffer solutions. In pH measure-
The required reagents are described in the section, Re-
ments, standard buffer solutions are required for reference
agents. Previously dry the crystalline reagents, except the
purposes. For convenience, the preparation of these solu-
boric acid and sodium acetate trihydrate, at 110 to 120
tions is in some instances described in the sections in which
for 1 hour.
their use is specified; i.e., five separate phosphate buffers are
[NOTEWhere water is specified for solution or dilution
described under AntibioticsMicrobial Assays 81, and sev-
of test substances in pH determinations, use carbon dioxide-
eral miscellaneous single-purpose solutions are described in
free water.]
the individual monographs.
Store the prepared solutions in chemically resistant, tight
A solution is said to be buffered if it resists changes in the
containers such as Type I glass bottles. Use the solutions
activity of an ion on the addition of substances that are
within 3 months.
expected to change the activity of that ion. Buffers are sub-
Standard Buffer Solutions for various ranges between pH
stances or combinations of substances that impart this resis-
1.2 and 10.0 may be prepared by appropriate combinations
tance to a solution. Buffered solutions are systems in which
of the solutions described herein, used in the proportions
the ion is in equilibrium with substances capable of remov-
shown in the accompanying table. The volumes shown in
ing or releasing the ion.
the table are for 200 mL of buffer solution, except that the
Buffer capacity refers to the amount of material that may
volumes shown for Acetate Buffer are used to prepare
be added to a solution without causing a significant change
1000 mL of buffer solution.
in ion activity. It is defined as the ratio of acid or base
1. Hydrochloric Acid, 0.2 M, and Sodium Hydroxide, 0.2
added (in gram-equivalents per liter) to the change in pH
MPrepare and standardize as directed under Volu-
(in pH units). The capacity of a buffered solution is adjusted
metric Solutions.
to the conditions of use, usually by adjustment of the con-
2. Potassium Biphthalate, 0.2 MDissolve 40.85 g of po-
centrations of buffer substances.
tassium biphthalate [KHC6H4(COO)2] in water, and di-
Buffers are used to establish and maintain an ion activity
lute with water to 1000 mL.
within narrow limits. The most common systems are used
3. Potassium Phosphate, Monobasic 0.2 MDissolve
(a) to establish hydrogen-ion activity for the calibration of
27.22 g of monobasic potassium phosphate (KH2PO4)
pH meters, (b) in the preparation of dosage forms that ap-
in water, and dilute with water to 1000 mL.
proach isotonicity, (c) in analytical procedures, and (d) to
4. Boric Acid and Potassium Chloride, 0.2 MDissolve
maintain stability of various dosage forms. Buffers used in
12.37 g of boric acid (H3BO3) and 14.91 g of potas-
physiological systems are carefully chosen so as not to inter-
sium chloride (KCl) in water, and dilute with water to
fere with pharmacological activity of the medicament or
1000 mL.
normal function of the organism. It is essential that buffers
5. Potassium Chloride, 0.2 MDissolve 14.91 g of potas-
used in chemical analysis be compatible with the substance
sium chloride (KCl) in water, and dilute with water to
determined and the reagents used.
1000 mL.
Standard Buffer SolutionsStandard solutions of definite
6. Acetic Acid, 2 NPrepare and standardize as directed
pH are readily available in buffer solutions prepared from
under Volumetric Solutions.
the appropriate reagents. In addition, buffer solutions, buffer
Composition of Standard Buffer Solutions
Hydrochloric Acid Buffer
Place 50 mL of the potassium chloride solution in a 200-mL volumetric flask, add the specified volume of the hydrochloric acid solution, then add water
to volume.
pH 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2.0 2.1 2.2
0.2 M HCl, mL 85.0 67.2 53.2 41.4 32.4 26.0 20.4 16.2 13.0 10.2 7.8
Acid Phthalate Buffer
Place 50 mL of the potassium biphthalate solution in a 200-mL volumetric flask, add the specified volume of the hydrochloric acid solution, then
add water to volume.
pH 2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6 3.8 4.0
0.2 M HCl, mL 49.5 42.2 35.4 28.9 22.3 15.7 10.4 6.3 2.9 0.1
Neutralized Phthalate Buffer
Place 50 mL of the potassium biphthalate solution in a 200-mL volumetric flask, add the specified volume of the sodium hydroxide solution, then
add water to volume.
pH 4.2 4.4 4.6 4.8 5.0 5.2 5.4 5.6 5.8
0.2 M NaOH, mL 3.0 6.6 11.1 16.5 22.6 28.8 34.1 38.8 42.3
1210 Buffer Solutions / Solutions USP 36
Phosphate Buffer
Place 50 mL of the monobasic potassium phosphate solution in a 200-mL volumetric flask, add the specified volume of the sodium hydroxide
solution, then add water to volume.
pH 5.8 6.0 6.2 6.4 6.6 6.8 7.0 7.2 7.4 7.6 7.8 8.0
0.2 M NaOH, mL 3.6 5.6 8.1 11.6 16.4 22.4 29.1 34.7 39.1 42.4 44.5 46.1
Alkaline Borate Buffer
Place 50 mL of the boric acid and potassium chloride solution in a 200-mL volumetric flask, add the specified volume of the sodium hydroxide
solution, then add water to volume.
pH 8.0 8.2 8.4 8.6 8.8 9.0 9.2 9.4 9.6 9.8 10.0
0.2 M NaOH, mL 3.9 6.0 8.6 11.8 15.8 20.8 26.4 32.1 36.9 40.6 43.7
Acetate Buffer
Place the specified amount of sodium acetate NaC2H3O2 3H2O in a 1000-mL volumetric flask, add the specified volume of the acetic acid solution,
then add water to volume, and mix.
pH 4.1 4.3 4.5 4.7 4.9 5.1 5.2 5.3 5.4 5.5
pH (measured) 4.10 4.29 4.51 4.70 4.90 5.11 5.18 5.30 5.40 5.48
NaC2H3O2 3H2O, g 1.5 1.99 2.99 3.59 4.34 5.08 5.23 5.61 5.76 5.98
2 N CH3COOH, mL 19.5 17.7 14.0 11.8 9.1 6.3 5.8 4.4 3.8 3.0
Ferric Chloride CSDissolve about 55 g of ferric chloride
COLORIMETRIC SOLUTIONS (CS)
(FeCl3 6H2O) in enough of a mixture of 25 mL of hydro-
chloric acid and 975 mL of water to make 1000 mL. Pipet
(For the Preparation of Matching Fluids, see Color and
10 mL of this solution into a 250-mL iodine flask, add 15 mL
Achromicity 631.)
of water, 3 g of potassium iodide, and 5 mL of hydrochloric
These solutions are used in the preparation of the colori-
acid, and allow the mixture to stand for 15 minutes. Dilute
metric standards for certain drugs, and for the carbonization
with 100 mL of water, and titrate the liberated iodine with
tests with sulfuric acid that are specified in several mono-
0.1 N sodium thiosulfate VS, adding 3 mL of starch TS as
graphs. Store the solutions in suitably resistant, tight
the indicator. Perform a blank determination with the same
containers.
quantities of the same reagents, and make any necessary
Comparison of colors as directed in the Pharmacopeial
correction. Each mL of 0.1 N sodium thiosulfate is equiva-
tests preferably is made in matched color-comparison tubes
lent to 27.03 mg of FeCl3 6H2O. Adjust the final volume of
or in a suitable colorimeter under conditions that ensure
the solution by the addition of enough of the mixture of
that the colorimetric reference solution and that of the spec-
hydrochloric acid and water so that each mL contains
imen under test are treated alike in all respects. The com-
45.0 mg of FeCl3 6H2O.
parison of colors is best made in layers of equal depth, and
viewed transversely against a white background (see also
Visual Comparison under Spectrophotometry and Light-Scatter-
INDICATOR SOLUTIONS
ing 851). It is particularly important that the solutions be
compared at the same temperature, preferably 25.
See TEST SOLUTIONS.
Cobaltous Chloride CSDissolve about 65 g of cobaltous
chloride (CoCl2 6H2O) in enough of a mixture of 25 mL of
TEST SOLUTIONS (TS) hydrochloric acid and 975 mL of water to make 1000 mL.
Pipet 5 mL of this solution into a 250-mL iodine flask, add
Certain of the following test solutions are intended for use 5 mL of hydrogen peroxide TS and 15 mL of sodium hy-
as acid-base indicators in volumetric analyses. Such solutions droxide solution (1 in 5), boil for 10 minutes, cool, and add
should be so adjusted that when 0.15 mL of the indicator 2 g of potassium iodide and 20 mL of dilute sulfuric acid (1
solution is added to 25 mL of carbon dioxide-free water, in 4). When the precipitate has dissolved, titrate the liber-
0.25 mL of 0.02 N acid or alkali, respectively, will produce ated iodine with 0.1 N sodium thiosulfate VS, adding 3 mL
the characteristic color change. Similar solutions are in- of starch TS as the indicator. Perform a blank determination
tended for use in pH measurement. Where no special direc- with the same quantities of the same reagents, and make
tions for their preparation are given, the same solution is any necessary correction. Each mL of 0.1 N sodium thiosul-
suitable for both purposes. fate is equivalent to 23.79 mg of CoCl2 6H2O. Adjust the
Where it is directed that a volumetric solution be used as final volume of the solution by the addition of enough of
the test solution, standardization of the solution used as TS the mixture of hydrochloric acid and water so that each mL
is not required. contains 59.5 mg of CoCl2 6H2O.
In general, the directive to prepare a solution fresh indi- Cupric Sulfate CSDissolve about 65 g of cupric sulfate
cates that the solution is of limited stability and must be (CuSO4 5H2O) in enough of a mixture of 25 mL of hydro-
prepared on the day of use. chloric acid and 975 mL of water to make 1000 mL. Pipet
For the preparation of Test Solutions, use reagents of the 10 mL of this solution into a 250-mL iodine flask, add 40 mL
quality described under Reagents. of water, 4 mL of acetic acid, 3 g of potassium iodide, and
Acetaldehyde TSMix 4 mL of acetaldehyde, 3 mL of 5 mL of hydrochloric acid, and titrate the liberated iodine
alcohol, and 1 mL of water. Prepare this solution fresh. with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS
Acetate Buffer TSDissolve 320 g of ammonium acetate as the indicator. Perform a blank determination with the
in 500 mL of water, add 5 mL of glacial acetic acid, dilute same quantities of the same reagents, and make any neces-
with water to 1000.0 mL, and mix. This solution has a pH sary correction. Each mL of 0.1 N sodium thiosulfate is
between 5.9 and 6.0. equivalent to 24.97 mg of CuSO4 5H2O. Adjust the final
Acetic Acid, Glacial, TSDetermine the water content volume of the solution by the addition of enough of the
of a specimen of glacial acetic acid by the Titrimetric Method mixture of hydrochloric acid and water so that each mL
(see Water Determination 921). If the acid contains more contains 62.4 mg of CuSO4 5H2O.
than 0.4% of water, add a few mL of acetic anhydride, mix,
allow to stand overnight, and again determine the water
USP 36 Solutions / Test Solutions 1211
content. If the acid contains less than 0.02% of water, add monia. Specific gravity: about 0.80. It contains between 9%
sufficient water to make the final concentration between and 11% of NH3. Store it in alkali-resistant containers, in a
0.02% and 0.4%, mix, allow to stand overnight, and again cold place.
determine the water content. Repeat the adjustment with Ammonia TS, StrongerUse Ammonia Water, Stronger
acetic anhydride or water, as necessary, until the resulting (see in the section Reagents).
solution shows a water content of not more than 0.4%. AmmoniaAmmonium Chloride Buffer TSDissolve
Acetic Acid, Strong, TSAdd 300.0 mL of glacial acetic 67.5 g of ammonium chloride in water, add 570 mL of am-
acid, and dilute with water to 1000 mL. This solution con- monium hydroxide, and dilute with water to 1000 mL.
tains about 30% (v/v) of CH3COOH and has a concentration AmmoniaCyanide TSDissolve 2 g of potassium cya-
of about 5 N. nide in 15 mL of ammonium hydroxide, and dilute with
Acetic AcidAmmonium Acetate Buffer TSDissolve water to 100 mL.
77.1 g of ammonium acetate in water, add 57 mL of glacial Ammoniacal Potassium Ferricyanide TSDissolve 2 g
acetic acid, and dilute with water to 1000 mL. of potassium ferricyanide in 75 mL of water, add 25 mL of
Acetone, Buffered, TSDissolve 8.15 g of sodium ace- ammonium hydroxide, and mix.
tate and 42 g of sodium chloride in about 100 mL of water, Ammoniated Cupric Oxide TSSee Cupric Oxide, Am-
and add 68 mL of 0.1 N hydrochloric acid and 150 mL of moniated, TS.
acetone. Mix, and dilute with water to 500 mL. Ammonium Acetate TSDissolve 10 g of ammonium
Acid Ferric Chloride TSMix 60 mL of glacial acetic acetate in water to make 100 mL.
acid with 5 mL of sulfuric acid, add 1 mL of ferric chloride Ammonium Carbonate TSDissolve 20 g of ammonium
TS, mix, and cool. carbonate and 20 mL of ammonia TS in water to make
Acid Ferrous Sulfate TSSee Ferrous Sulfate, Acid, TS. 100 mL.
Acid Stannous Chloride TSSee Stannous Chloride, Acid, Ammonium Carbonate TS 2Prepare a 158-mg/mL so-
TS. lution of ammonium carbonate in water.
Ammonium Chloride TSDissolve 10.5 g of ammonium
chloride in water to make 100 mL.
Change to read:
Ammonium ChlorideAmmonium Hydroxide TSMix
equal volumes of water and ammonium hydroxide, and sat-
Acid Stannous Chloride TS, StrongerSee
Stannous
urate with ammonium chloride.
Chloride, Acid, Stronger, TS.
(ERR 1-Jul-2012)
Ammonium Molybdate TSDissolve 6.5 g of finely
Albumen TSCarefully separate the white from the yolk
powdered molybdic acid in a mixture of 14 mL of water
of a strictly fresh hens egg. Shake the white with 100 mL of
and 14.5 mL of ammonium hydroxide. Cool the solution,
water until mixed and all but the chalaza has undergone
and add it slowly, with stirring, to a well-cooled mixture of
solution; then filter. Prepare the solution fresh.
32 mL of nitric acid and 40 mL of water. Allow to stand for
AlcoholPhenol TSDissolve 780 mg of phenol in alco-
48 hours, and filter through a fine-porosity, sintered-glass
hol to make 100 mL.
crucible. This solution deteriorates upon standing and is un-
Alcoholic TSIt contains 95 parts of specially
suitable for use if, upon the addition of 2 mL of dibasic so-
denaturated alcohol 3A with 5 parts of isopropyl alcohol.
dium phosphate TS to 5 mL of the solution, an abundant
The final concentrations are approximately 90% alcohol, 5%
yellow precipitate does not form at once or after slight
methanol, and 5% isopropanol.
warming. Store it in the dark. If a precipitate forms during
[NOTEA suitable grade is available as Reagent alcohol,
storage, use only the clear supernatant.
catalog number R8382, available at www.sigma-aldrich.
Ammonium Oxalate TSDissolve 3.5 g of ammonium
com.]
oxalate in water to make 100 mL.
Alcoholic Ammonia TSSee Ammonia TS, Alcoholic.
Ammonium Phosphate, Dibasic, TS (Ammonium
Alcoholic Mercuric Bromide TSSee Mercuric Bromide
Phosphate TS)Dissolve 13 g of dibasic ammonium phos-
TS, Alcoholic.
phate in water to make 100 mL.
Alcoholic Potassium Hydroxide TSSee Potassium Hy-
Ammonium Polysulfide TSYellow liquid, made by sat-
droxide TS, Alcoholic.
urating ammonium sulfide TS with sulfur.
Alkaline Cupric Citrate TSSee Cupric Citrate TS, Alka-
Ammonium Pyrrolidinedithiocarbamate, Saturated,
line.
TSAdd about 10 g of ammonium pyrrolidinedithiocarba-
Alkaline Cupric Citrate TS 2See Cupric Citrate TS 2,
mate to a 1000-mL volumetric flask, and dilute with water
Alkaline.
to volume.
Alkaline Cupric Iodide TSSee Cupric Iodide TS, Alkaline.
Ammonium Reineckate TSShake about 500 mg of
Alkaline Cupric Tartrate TS (Fehlings Solution)See Cu-
ammonium reineckate with 20 mL of water frequently dur-
pric Tartrate TS, Alkaline.
ing 1 hour, and filter. Use within 2 days.
Alkaline MercuricPotassium Iodide TSSee
Ammonium Sulfide TSUse ACS reagent-grade Ammo-
MercuricPotassium Iodide TS, Alkaline.
nium Sulfide Solution.
Alkaline Picrate TSSee Picrate TS, Alkaline.
Ammonium Thiocyanate TSDissolve 8 g of ammo-
Alkaline Sodium Hydrosulfite TSSee Sodium Hydrosul-
nium thiocyanate in water to make 100 mL.
fite TS, Alkaline.
Ammonium Vanadate TSDissolve 2.5 g of ammonium
Amaranth TSDissolve 20 mg of amaranth in 10 mL of
vanadate in 500 mL of boiling water, cool, and add 20 mL
water.
of nitric acid. Mix, cool, and add water to make 1 L. Store
Aminonaphtholsulfonic Acid TSAccurately weigh 5 g
in polyethylene containers.
of sodium sulfite, 94.3 g of sodium bisulfite, and 700 mg of
Anthrone TSWithin 12 hours of use, rapidly dissolve
1,2,4-aminonaphtholsulfonic acid, and mix. Prepare amino-
35 mg of anthrone in a hot mixture of 35 mL of water and
naphtholsulfonic acid TS fresh on the day of use by dissolv-
65 mL of sulfuric acid. Immediately cool in an ice bath to
ing 1.5 g of the dry mixture in 10 mL of water.
room temperature, and filter through glass wool. Allow the
Ammonia TSIt contains between 9.5% and 10.5% of
solution to stand at room temperature for 30 minutes before
NH3. Prepare by diluting 350 mL of Ammonia Water,
use.
Stronger (see in the section, Reagents) with water to make
Antimony Trichloride TSDissolve 20 g of antimony
1000 mL.
trichloride in chloroform to make 100 mL. Filter if necessary.
Ammonia TS 2Prepare by diluting 13.5 mL of Ammo-
Barium Chloride TSDissolve 12 g of barium chloride in
nia Water, Stronger (see Reagent Specifications in the section
water to make 100 mL.
Reagents) with water to make 100 mL.
Barium Hydroxide TSA saturated solution of barium
Ammonia TS, AlcoholicA solution of ammonia gas in
hydroxide in recently boiled water. Prepare the solution
alcohol. Clear, colorless liquid having a strong odor of am-
fresh.
1212 Test Solutions / Solutions USP 36
Barium Nitrate TSDissolve 6.5 g of barium nitrate in Cobaltous Chloride TSDissolve 2 g of cobaltous chlo-
water to make 100 mL. ride in 1 mL of hydrochloric acid and sufficient water to
Betanaphthol TSSee 2-Naphthol TS. make 100 mL.
Biuret Reagent TSDissolve 1.5 g of cupric sulfate and Congo Red TSDissolve 500 mg of congo red in a mix-
6.0 g of potassium sodium tartrate in 500 mL of water in a ture of 10 mL of alcohol and 90 mL of water.
1000-mL volumetric flask. Add 300 mL of carbonate-free so- m-Cresol Purple TSDissolve 0.10 g of metacresol pur-
dium hydroxide solution (1 in 10), dilute with carbonate- ple in 13 mL of 0.01 N sodium hydroxide, dilute with water
free sodium hydroxide solution (1 in 10) to 1000 mL, and to 100 mL, and mix.
mix. Cresol Red TSTriturate 100 mg of cresol red in a mor-
Blue Tetrazolium TSDissolve 500 mg of blue tet- tar with 26.2 mL of 0.01 N sodium hydroxide until solution
razolium in alcohol to make 100 mL. is complete, then dilute the solution with water to 250 mL.
Brilliant Blue G TSTransfer 25 mg of brilliant blue G to Cresol RedThymol Blue TSAdd 15 mL of thymol blue
a 100-mL volumetric flask, add 12.5 mL of alcohol and TS to 5 mL of cresol red TS, and mix.
25 mL of phosphoric acid, dilute with water to volume, and Crystal Violet TSDissolve 100 mg of crystal violet in
mix. 10 mL of glacial acetic acid.
Bromine TS (Bromine Water)A saturated solution of Cupric Acetate TSDissolve 100 mg of cupric acetate in
bromine, prepared by agitating 2 to 3 mL of bromine with about 5 mL of water to which a few drops of acetic acid
100 mL of cold water in a glass-stoppered bottle, the stop- have been added. Dilute to 100 mL, and filter, if necessary.
per of which should be lubricated with petrolatum. Store it Cupric Acetate TS, Stronger (Barfoeds Reagent)Dis-
in a cold place, protected from light. solve 13.3 g of cupric acetate in a mixture of 195 mL of
BromineSodium Acetate TSDissolve 100 g of sodium water and 5 mL of acetic acid.
acetate in 1000 mL of glacial acetic acid, add 50 mL of bro- Cupric-Ammonium Sulfate TSTo cupric sulfate TS add
mine, and mix. ammonia TS, dropwise, until the precipitate initially formed
p-Bromoaniline TSAdd 8 g of p-bromoaniline to a is nearly but not completely dissolved. Allow to settle, and
mixture of 380 mL of thiourea-saturated glacial acetic acid, decant the clear solution. Prepare this solution fresh.
10 mL of sodium chloride solution (1 in 5), 5 mL of oxalic Cupric Citrate TSDissolve 25 g of cupric sulfate, 50 g
acid solution (1 in 20), and 5 mL of dibasic sodium phos- of citric acid, and 144 g of anhydrous sodium carbonate in
phate solution (1 in 10) in a low-actinic glass bottle. Mix, water, and dilute with water to 1000 mL.
and allow to stand overnight before using. Protect from Cupric Citrate TS, AlkalineWith the aid of heat, dis-
light, and use within 7 days. solve 173 g of dihydrated sodium citrate and 117 g of mon-
Bromocresol Blue TSUse Bromocresol Green TS. ohydrated sodium carbonate in about 700 mL of water, and
Bromocresol Green TSDissolve 50 mg of bromocresol filter through paper, if necessary, to obtain a clear solution.
green in 100 mL of alcohol, and filter if necessary. In a separate container dissolve 17.3 g of cupric sulfate in
Bromocresol GreenMethyl Red TSDissolve 0.15 g of about 100 mL of water, and slowly add this solution, with
bromocresol green and 0.1 g of methyl red in 180 mL of constant stirring, to the first solution. Cool the mixture, add
alcohol, and dilute with water to 200 mL. water to make 1000 mL, and mix.
Bromocresol Purple TSDissolve 250 mg of bromo- Cupric Citrate TS 2, AlkalineWith the aid of heat, dis-
cresol purple in 20 mL of 0.05 N sodium hydroxide, and solve about 173 g of sodium citrate dihydrate and 117 g of
dilute with water to 250 mL. sodium carbonate monohydrate in about 700 mL of water,
Bromophenol Blue TSDissolve 100 mg of bromophe- and filter. In a second flask, dissolve about 27.06 g of cupric
nol blue in 100 mL of diluted alcohol, and filter if necessary. sulfate (CuSO4 5H2O) in about 100 mL of water. Slowly
Bromothymol Blue TSDissolve 100 mg of combine the two solutions while stirring, and dilute with
bromothymol blue in 100 mL of diluted alcohol, and filter if water to 1000 mL.
necessary. Cupric Iodide TS, AlkalineDissolve 7.5 g of cupric sul-
Buffered Acetone TSSee Acetone, Buffered, TS. fate (CuSO4 5H2O) in about 100 mL of water. In a separate
Calcium Chloride TSDissolve 7.5 g of calcium chloride container dissolve 25 g of anhydrous sodium carbonate,
in water to make 100 mL. 20 g of sodium bicarbonate, and 25 g of potassium sodium
Calcium Hydroxide TSUse Calcium Hydroxide Topical tartrate in about 600 mL of water. With constant stirring,
Solution (USP monograph). add the cupric sulfate solution to the bottom of the alkaline
Calcium Sulfate TSA saturated solution of calcium sul- tartrate solution by means of a funnel that touches the bot-
fate in water. tom of the container. Add 1.5 g of potassium iodide, 200 g
Ceric Ammonium Nitrate TSDissolve 6.25 g of ceric of anhydrous sodium sulfate, 50 to 150 mL of 0.02 M po-
ammonium nitrate in 10 mL of 0.25 N nitric acid. Use tassium iodate, and sufficient water to make 1000 mL.
within 3 days. Cupric Oxide, Ammoniated, TS (Schweitzers Reagent)
Chloral Hydrate TSDissolve 50 g of chloral hydrate in Dissolve 10 g of cupric sulfate in 100 mL of water, add suffi-
a mixture of 15 mL of water and 10 mL of glycerin. cient sodium hydroxide solution (1 in 5) to precipitate the
Chlorine TS (Chlorine Water)A saturated solution of copper hydroxide, collect the latter on a filter, and wash
chlorine in water. Place the solution in small, completely free from sulfate with cold water. Dissolve the precipitate,
filled, light-resistant containers. Chlorine TS, even when kept which must be kept wet during the entire process, in the
from light and air, is apt to deteriorate. Store it in a cold, minimum quantity of ammonia TS necessary for complete
dark place. For full strength, prepare this solution fresh. solution.
Chromotropic Acid TSDissolve 50 mg of chromotropic Cupric Sulfate TSDissolve 12.5 g of cupric sulfate in
acid or its disodium salt in 100 mL of 75% sulfuric acid, water to make 100 mL.
which may be made by cautiously adding 75 mL of sulfuric Cupric Tartrate TS, Alkaline (Fehlings Solution)
acid to 33.3 mL of water.
The Copper Solution (A)Dissolve 34.66 g of carefully se-
CobaltUranyl Acetate TSDissolve, with warming,
lected, small crystals of cupric sulfate, showing no trace of
40 g of uranyl acetate in a mixture of 30 g of glacial acetic
efflorescence of adhering moisture, in water to make
acid and sufficient water to make 500 mL. Similarly, prepare
500 mL. Store this solution in small, tight containers.
a solution containing 200 g of cobaltous acetate in a mix-
The Alkaline Tartrate Solution (B)Dissolve 173 g of crys-
ture of 30 g of glacial acetic acid and sufficient water to
tallized potassium sodium tartrate and 50 g of sodium hy-
make 500 mL. Mix the two solutions while still warm, and
droxide in water to make 500 mL. Store this solution in
cool to 20. Maintain the temperature at 20 for about
small, alkali-resistant containers.
2 hours to separate the excess salts from solution, and then
For use, mix exactly equal volumes of Solutions A and B at
pass through a dry filter.
the time required.
Delafields Hematoxylin TSPrepare 400 mL of a satu- Fehlings SolutionSee Cupric Tartrate TS, Alkaline.
rated solution of ammonium alum (Solution A). Dissolve 4 g Ferric Ammonium Sulfate TSDissolve 8 g of ferric am-
of hematoxylin in 25 mL of alcohol, mix it with Solution A, monium sulfate in water to make 100 mL.
and allow it to stand for 4 days in a flask closed with a Ferric Chloride TSDissolve 9 g of ferric chloride in
pledget of purified cotton and exposed to light and air (So- water to make 100 mL.
lution B). Then filter Solution B, and add to it a Solution C Ferroin TSDissolve 0.7 g of ferrous sulfate and 1.76 g
consisting of a mixture of 100 mL of glycerin and 100 mL of of o-phenanthroline monohydrochloride monohydrate in
methanol. Mix, and allow the mixture to stand in a warm water, and dilute with water to 100 mL.
place, exposed to light, for 6 weeks until it becomes dark- Ferrous Sulfate TSDissolve 8 g of clear crystals of fer-
colored. Store in tightly stoppered bottles. rous sulfate in about 100 mL of recently boiled and thor-
For use in staining endocrine tissue, dilute this test solu- oughly cooled water. Prepare this solution fresh.
tion with an equal volume of water. Ferrous Sulfate, Acid, TSDissolve 7 g of ferrous sulfate
Denaturated Alcoholic TSA specially denaturated alco- crystals in 90 mL of recently boiled and thoroughly cooled
hol containing either rubber hydrocarbon solvent of hep- water, and add sulfuric acid to make 100 mL. Prepare this
tane or toluene. [NOTEA suitable grade is available from solution immediately prior to use.
www.lyondell.com or from www.sasol.com, as Ethanol SDA Folin-Ciocalteu Phenol TSInto a 1500-mL flask intro-
2B HEP 200, or Ethanol SDA 2B TOL 200, or Ethanol SDA duce 100 g of sodium tungstate, 25 g of sodium molybdate,
2B TOL 190, or Alcohol SDA 2B-2.] 700 mL of water, 50 mL of phosphoric acid, and 100 mL of
Denig` es ReagentSee Mercuric Sulfate TS. hydrochloric acid. Gently reflux the mixture for about
Diazobenzenesulfonic Acid TSPlace in a beaker 1.57 g 10 hours, and add 150 g of lithium sulfate, 50 mL of water,
of sulfanilic acid, previously dried at 105 for 3 hours, add and a few drops of bromine. Boil the mixture, without the
80 mL of water and 10 mL of diluted hydrochloric acid, and condenser, for about 15 minutes, or until the excess bro-
warm on a steam bath until dissolved. Cool to 15 (some of mine is expelled. Cool, dilute with water to 1 L, and filter:
the sulfanilic acid may separate but will dissolve later), and the filtrate has no greenish tint. Before use, dilute 1 part of
add slowly, with constant stirring, 6.5 mL of sodium nitrite the filtrate with 1 part of water. When used for protein de-
solution (1 in 10). Then dilute with water to 100 mL. termination (i.e., Lowry assay), this reagent must be further
Dichlorofluorescein TSDissolve 100 mg of dichloro- diluted (1:5) with water. See Method 2 in Total Protein Assay
fluorescein in 60 mL of alcohol, add 2.5 mL of 0.1 N sodium under Biotechnology-Derived ArticlesTotal Protein Assay
hydroxide, mix, and dilute with water to 100 mL. 1057.
2,7-Dihydroxynaphthalene TSDissolve 100 mg of 2,7- Formaldehyde TSUse Formaldehyde Solution (see in the
dihydroxynaphthalene in 1000 mL of sulfuric acid, and allow section Reagents).
the solution to stand until the yellow color disappears. If the FuchsinPyrogallol TSDissolve 100 mg of basic fuchsin
solution is very dark, discard it and prepare a new solution in 50 mL of water that previously has been boiled for
from a different supply of sulfuric acid. This solution is stable 15 minutes and allowed to cool slightly. Cool, add 2 mL of
for approximately 1 month if stored in a dark bottle. a saturated solution of sodium bisulfite, mix, and allow to
Diiodofluorescein TSDissolve 500 mg of stand for not less than 3 hours. Add 0.9 mL of hydrochloric
diiodofluorescein in a mixture of 75 mL of alcohol and acid, mix, and allow to stand overnight. Add 100 mg of
30 mL of water. pyrogallol, shake until solution is effected, and dilute with
Diluted Lead Subacetate TSSee Lead Subacetate TS, water to 100 mL. Store in an amber-colored glass bottle in a
Diluted. refrigerator.
p-Dimethylaminobenzaldehyde TSDissolve 125 mg of FuchsinSulfurous Acid TSDissolve 200 mg of basic
p-dimethylaminobenzaldehyde in a cooled mixture of 65 mL fuchsin in 120 mL of hot water, and allow the solution to
of sulfuric acid and 35 mL of water, and add 0.05 mL of cool. Add a solution of 2 g of anhydrous sodium sulfite in
ferric chloride TS. Use within 7 days. 20 mL of water, then add 2 mL of hydrochloric acid. Dilute
Dinitrophenylhydrazine TSCarefully mix 10 mL of the solution with water to 200 mL, and allow to stand for at
water and 10 mL of sulfuric acid, and cool. To the mixture, least 1 hour. Prepare this solution fresh.
contained in a glass-stoppered flask, add 2 g of 2,4-dini- Gastric Fluid, Simulated, TSDissolve 2.0 g of sodium
trophenylhydrazine, and shake until dissolved. To the solu- chloride and 3.2 g of purified pepsin, that is derived from
tion add 35 mL of water, mix, cool, and filter. porcine stomach mucosa, with an activity of 800 to 2500
Diphenylamine TSDissolve 1.0 g of diphenylamine in units per mg of protein, in 7.0 mL of hydrochloric acid and
100 mL of sulfuric acid. The solution should be colorless. sufficient water to make 1000 mL. [NOTEPepsin activity is
Diphenylcarbazone TSDissolve 1 g of crystalline described in the Food Chemicals Codex specifications under
diphenylcarbazone in 75 mL of alcohol, then add alcohol to General Tests and Assays.] This test solution has a pH of
make 100 mL. Store in a brown bottle. about 1.2.
Dithizone TSDissolve 25.6 mg of dithizone in 100 mL Gelatin TS (for the assay of Corticotropin Injection)Dis-
of alcohol. Store in a cold place, and use within 2 months. solve 340 g of acid-treated precursor gelatin (Type A) in
Dragendorffs TSMix 850 mg of bismuth subnitrate water to make 1000 mL. Heat the solution in an autoclave
with 40 mL of water and 10 mL of glacial acetic acid (Solu- at 115 for 30 minutes after the exhaust line temperature
tion A). Dissolve 8 g of potassium iodide in 20 mL of water has reached 115. Cool the solution, and add 10 g of phe-
(Solution B). Mix equal portions of Solution A and Solution B nol and 1000 mL of water. Store in tight containers in a
to obtain a stock solution, which can be stored for several refrigerator.
months in a dark bottle. Mix 10 mL of the stock solution Glacial Acetic Acid TSSee Acetic Acid, Glacial, TS.
with 20 mL of glacial acetic acid, and dilute with water to Glucose OxidaseChromogen TSA solution contain-
make 100 mL. ing, in each mL, 0.5 mol of 4-aminoantipyrine, 22.0 mol
Edetate Disodium TSDissolve 1 g of edetate disodium of sodium p-hydroxybenzoate, not less than 7.0 units of glu-
in 950 mL of water, add 50 mL of alcohol, and mix. cose oxidase, and not less than 0.5 units of peroxidase, and
Eosin Y TS (adsorption indicator)Dissolve 50 mg of eo- buffered to a pH of 7.0 0.1.
sin Y in 10 mL of water.
SuitabilityWhen used for determining glucose in Inulin,
Eriochrome Black TSDissolve 200 mg of eriochrome
ascertain that no significant color results by reaction with
black T and 2 g of hydroxylamine hydrochloride in metha-
fructose, and that a suitable absorbance-versus-concentra-
nol to make 50 mL.
tion slope is obtained with glucose.
Eriochrome Cyanine TSDissolve 750 mg of eriochrome
[NOTEA suitable grade is available, as a concentrate,
cyanine R in 200 mL of water, add 25 g of sodium chloride,
from Worthington Diagnostics, Division of Millipore Corp.,
25 g of ammonium nitrate, and 2 mL of nitric acid, and di-
www.millipore.com.]
lute with water to 1000 mL.
Glycerin Base TSTo 200 g of glycerin add water to IronPhenol TS (Iron-Kober Reagent)Dissolve 1.054 g of
bring the total weight to 235 g. Add 140 mL of 1 N sodium ferrous ammonium sulfate in 20 mL of water, and add 1 mL
hydroxide and 50 mL of water. of sulfuric acid and 1 mL of 30 percent hydrogen peroxide.
Gold Chloride TSDissolve 1 g of gold chloride in Mix, heat until effervescence ceases, and dilute with water
35 mL of water. to 50 mL. To 3 volumes of this solution contained in a volu-
Hydrogen Peroxide TSUse Hydrogen Peroxide Topical metric flask add sulfuric acid, with cooling, to make 100
Solution (USP monograph). volumes. Purify phenol by distillation, discarding the first
Hydrogen Sulfide TSA saturated solution of hydrogen 10% and the last 5%, collecting the distillate, with exclusion
sulfide, made by passing H2S into cold water. Store it in of moisture, in a dry, tared glass-stoppered flask of about
small, dark amber-colored bottles, filled nearly to the top. It twice the volume of the phenol. Solidify the phenol in an
is unsuitable unless it possesses a strong odor of H2S, and ice bath, breaking the top crust with a glass rod to ensure
unless it produces at once a copious precipitate of sulfur complete crystallization. Weigh the flask and its contents,
when added to an equal volume of ferric chloride TS. Store add to the phenol 1.13 times its weight of the ironsulfuric
in a cold, dark place. acid solution prepared as directed, insert the stopper in the
Hydroxylamine Hydrochloride TSDissolve 3.5 g of hy- flask, and allow to stand, without cooling but with occa-
droxylamine hydrochloride in 95 mL of 60% alcohol, and sional mixing, until the phenol is liquefied. Shake the mix-
add 0.5 mL of bromophenol blue solution (1 in 1000 of ture vigorously until mixed, allow to stand in the dark for
alcohol) and 0.5 N alcoholic potassium hydroxide until a 16 to 24 hours, and again weigh the flask and its contents.
greenish tint develops in the solution. Then add 60% alco- To the mixture add 23.5% of its weight of a solution of 100
hol to make 100 mL. volumes of sulfuric acid in 110 volumes of water, mix, trans-
8-Hydroxyquinoline TSDissolve 5 g of 8-hydroxy- fer to dry glass-stoppered bottles, and store in the dark,
quinoline in alcohol to make 100 mL. protected from atmospheric moisture. Use within 6 months.
Indigo Carmine TS (Sodium Indigotindisulfonate TS)Dis- Dispense the reagent from a small-bore buret, arranged to
solve a quantity of sodium indigotindisulfonate, equivalent exclude moisture, capable of delivering 1 mL in 30 seconds
to 180 mg of C16H8N2O2(SO3Na)2, in water to make 100 mL. or less, and having no lubricant, other than reagent, on its
Use within 60 days. stopcock. Wipe the buret tip with tissue before each addi-
IndophenolAcetate TS (for the assay of Corticotropin In- tion.
jection)To 60 mL of standard dichlorophenol-indophenol Iron Salicylate TSDissolve 500 mg of ferric ammonium
solution (see in the section Volumetric Solutions) add water sulfate in 250 mL of water containing 10 mL of diluted sul-
to make 250 mL. Add to the resulting solution an equal vol- furic acid, and add water to make 500 mL. To 100 mL of
ume of sodium acetate solution freshly prepared by dissolv- the resulting solution add 50 mL of a 1.15% solution of so-
ing 13.66 g of anhydrous sodium acetate in water to make dium salicylate, 20 mL of diluted acetic acid, and 80 mL of a
500 mL and adjusting with 0.5 N acetic acid to a pH of 7. 13.6% solution of sodium acetate, then add water to make
Store in a refrigerator, and use within 2 weeks. 500 mL. Store in a well-closed container. Protect from light.
Intestinal Fluid, Simulated, TSDissolve 6.8 g of mono- Use within 2 weeks.
basic potassium phosphate in 250 mL of water, mix, and Lanthanum Nitrate TSDissolve 5.0 g of lanthanum ni-
add 77 mL of 0.2 N sodium hydroxide and 500 mL of trate hexahydrate in 100 mL of water.
water. Add 10.0 g of pancreatin, mix, and adjust the result- Lead Acetate TSDissolve 9.5 g of clear, transparent
ing solution with either 0.2 N sodium hydroxide or 0.2 N crystals of lead acetate in recently boiled water to make
hydrochloric acid to a pH of 6.8 0.1. Dilute with water to 100 mL. Store in well-stoppered bottles.
1000 mL. Lead Acetate TS, AlcoholicDissolve 2 g of clear, trans-
Iodine TSUse 0.1 N Iodine (see in the section Volumet- parent crystals of lead acetate in alcohol to make 100 mL.
ric Solutions). Store in tight containers.
Iodine, Diluted TSTransfer 10.0 mL of 0.1 N iodine VS Lead Subacetate TSDissolve 40.0 g of lead acetate in
to a 100-mL volumetric flask, dilute with water to volume, 90 mL of carbon dioxide-free water. Adjust with 10 M so-
and mix. dium hydroxide to a pH of 7.5, centrifuge, and use the clear
Iodine Monochloride TSDissolve 10 g of potassium io- supernatant. It contains NLT 16.7% (w/w) and NMT 17.4%
dide and 6.44 g of potassium iodate in 75 mL of water in a (w/w) of Pb in a form corresponding to the formula
glass-stoppered container. Add 75 mL of hydrochloric acid C8H14O10Pb3. The solution remains clear when stored in a
and 5 mL of chloroform, and adjust to a faint iodine color well-closed container.
(in the chloroform) by adding dilute potassium iodide or Lead Subacetate TS, DilutedDilute 3.25 mL of lead
potassium iodate solution. If much iodine is liberated, use a subacetate TS with water, recently boiled and cooled, to
stronger solution of potassium iodate than 0.01 M at first, make 100 mL. Store in small, well-filled, tight containers.
making the final adjustment with the 0.01 M potassium io- Litmus TSDigest 25 g of powdered litmus with three
date. Store in a dark place, and readjust to a faint iodine successive 100-mL portions of boiling alcohol, continuing
color as necessary. each extraction for about 1 hour. Filter, wash with alcohol,
Iodine and Potassium Iodide TS 1Dissolve 500 mg of and discard the alcohol filtrate. Macerate the residue with
iodine and 1.5 g of potassium iodide in 25 mL of water. about 25 mL of cold water for 4 hours, filter, and discard
Iodine and Potassium Iodide TS 2Dissolve 12.7 g of the filtrate. Finally digest the residue with 125 mL of boiling
iodine and 20 g of potassium iodide in water, and dilute water for 1 hour, cool, and filter.
with water to 1000.0 mL. To 10.0 mL of this solution, add Locke-Ringers SolutionSee Locke-Ringers TS.
0.6 g of potassium iodide, and dilute with water to Locke-Ringers TS (Locke-Ringers Solution)
100.0 mL. Prepare immediately before use.
Iodine and Potassium Iodide TS 3Dissolve 0.127 g of
iodine and 0.20 g of potassium iodide in water, and dilute
Sodium Chloride 9.0 g
with water to 10.0 mL.
Potassium Chloride 0.42 g
Iodobromide TSDissolve 20 g of iodine monobromide
Calcium Chloride 0.24 g
in glacial acetic acid to make 1000 mL. Store in glass con-
tainers, protected from light. Magnesium Chloride 0.2 g
Iodochloride TSDissolve 16.5 g of iodine monochlo-
Sodium Bicarbonate 0.5 g
ride in 1000 mL of glacial acetic acid.
Dextrose 0.5 g
Iodoplatinate TSDissolve 300 mg of platinic chloride
Water, recently distilled from a hard-glass
in 97 mL of water. Immediately prior to use, add 3.5 mL of
flask, a sufficient quantity to make 1000 mL
potassium iodide TS, and mix.
Prepare fresh each day. The constituents (except the dex- Methyl Red TS 2To 1.86 mL of 0.1 M sodium hydrox-
trose and the sodium bicarbonate) may be made up in ide and 50 mL of alcohol, add 50 mg of methyl red, and
stock solutions and diluted as needed. dilute with water to 100 mL.
Magnesia Mixture TSDissolve 5.5 g of magnesium Methyl Red TS, MethanolicDissolve 1 g of methyl red
chloride and 7 g of ammonium chloride in 65 mL of water, in 100 mL of methanol, and filter, if necessary. Store pro-
add 35 mL of ammonia TS, set the mixture aside for a few tected from light, and use within 21 days.
days in a well-stoppered bottle, and filter. If the solution is Methyl RedMethylene Blue TSAdd 10 mL of methyl
not perfectly clear, filter it before using. red TS to 10 mL of methylene blue TS, and mix.
Magnesium Sulfate TSDissolve 12 g of crystals of Methyl Violet TSUse Crystal Violet TS.
magnesium sulfate, selected for freedom from efflorescence, Methyl Yellow TSPrepare a solution containing
in water to make 100 mL. 0.10 mg per mL in alcohol.
Malachite Green TSDissolve 1 g of malachite green Methyl YellowMethylene Blue TSDissolve 1 g of
oxalate in 100 mL of glacial acetic acid. methyl yellow and 100 mg of methylene blue in 125 mL of
Mallorys StainDissolve 500 mg of water-soluble ani- methanol.
line blue, 2 g of orange G, and 2 g of oxalic acid in 100 mL 3-Methyl-2-benzothiazolinone Hydrazone
of water. Hydrochloride TSDissolve 0.1 g of 3-methyl-2-benzo-
Mayers ReagentSee MercuricPotassium Iodide TS. thiazolinone hydrazone hydrochloride monohydrate in
Mercuric Acetate TSDissolve 6.0 g of mercuric acetate 10 mL of water, dilute the resulting solution with methanol
in glacial acetic acid to make 100 mL. Store in tight contain- to 100 mL, and mix.
ers, protected from direct sunlight. Methylene Blue TSDissolve 125 mg of methylene blue
MercuricAmmonium Thiocyanate TSDissolve 30 g of in 100 mL of alcohol, and dilute with alcohol to 250 mL.
ammonium thiocyanate and 27 g of mercuric chloride in Methylthionine Perchlorate TSTo 500 mL of potas-
water to make 1000 mL. sium perchlorate solution (1 in 1000) add dropwise, with
Mercuric Bromide TS, AlcoholicDissolve 5 g of mercu- constant shaking, methylene blue solution (1 in 100) until a
ric bromide in 100 mL of alcohol, employing gentle heat to slight, permanent turbidity results. Allow the precipitate to
facilitate solution. Store in glass containers, protected from settle, decant the supernatant through paper, and use only
light. the clear solution.
Mercuric Chloride TSDissolve 6.5 g of mercuric chlo- Millons ReagentTo 2 mL of mercury in a conical flask
ride in water to make 100 mL. add 20 mL of nitric acid. Shake the flask under a hood to
Mercuric Iodide TS (Valsers Reagent)Slowly add potas- break up the mercury into small globules. After about
sium iodide solution (1 in 10) to red mercuric iodide until 10 minutes, add 35 mL of water, and, if a precipitate or
almost all of the latter is dissolved, and filter off the excess. crystals appear, add sufficient dilute nitric acid (1 in 5, pre-
A solution containing 10 g of potassium iodide in 100 mL pared from nitric acid from which the oxides have been
dissolves approximately 14 g of HgI2 at 20. removed by blowing air through it until it is colorless) to
Mercuric Nitrate TSDissolve 40 g of mercuric oxide dissolve the separated solid. Add sodium hydroxide solution
(red or yellow) in a mixture of 32 mL of nitric acid and (1 in 10) dropwise, with thorough mixing, until the curdy
15 mL of water. Store in glass containers, protected from precipitate that forms after the addition of each drop no
light. longer redissolves but is dispersed to form a suspension.
MercuricPotassium Iodide TS (Mayers Reagent)Dis- Add 5 mL more of the dilute nitric acid, and mix. Prepare
solve 1.358 g of mercuric chloride in 60 mL of water. Dis- this solution fresh.
solve 5 g of potassium iodide in 10 mL of water. Mix the Molybdo-phosphotungstate TS (Folin-Denis Reagent)
two solutions, and dilute with water to 100 mL. To about 350 mL of water contained in a round-bottom
MercuricPotassium Iodide TS, Alkaline (Nesslers flask add 50 g of sodium tungstate, 12 g of phosphomolyb-
Reagent)Dissolve 143 g of sodium hydroxide in 700 mL of dic acid, and 25 mL of phosphoric acid. Boil the mixture
water. Dissolve 50 g of red mercuric iodide and 40 g of po- under a reflux condenser for 2 hours, then cool, dilute with
tassium iodide in 200 mL of water. Pour the iodide solution water to 500 mL, and mix. Store in tight containers, pro-
into the hydroxide solution, and dilute with water to tected from light, and in a cold place.
1000 mL. Allow to settle, and use the clear supernatant. 1-Naphthol ReagentDissolve 1 g of 1-naphthol in
Mercuric Sulfate TS (Denig` es Reagent)Mix 5 g of yel- 25 mL of methanol. Prepare this solution fresh.
low mercuric oxide with 40 mL of water, and while stirring 1-Naphthol TSUse 1-Naphthol Reagent.
slowly add 20 mL of sulfuric acid, then add another 40 mL 2-Naphthol TS (Betanaphthol TS)Dissolve 1 g of
of water, and stir until completely dissolved. 2-naphthol in 100 mL of sodium hydroxide solution (1 in
Mercurous Nitrate TSDissolve 15 g of mercurous ni- 100).
trate in a mixture of 90 mL of water and 10 mL of diluted p-Naphtholbenzein TSDissolve 250 mg of p-naphthol-
nitric acid. Store in dark, amber-colored bottles in which a benzein in 100 mL of glacial acetic acid.
small globule of mercury has been placed. N-(1-Naphthyl)ethylenediamine Dihydrochloride TS
Metaphenylenediamine Hydrochloride TSDissolve 1 g Dissolve 100 mg of N-(1-naphthyl)ethylenediamine dihydro-
of metaphenylenediamine hydrochloride in 200 mL of chloride in 100 mL of a mixture of 7 parts of acetone and
water. The solution must be colorless when used. If neces- 3 parts of water.
sary, decolorize by heating with activated charcoal. Nesslers ReagentSee MercuricPotassium Iodide TS, Al-
MetaphosphoricAcetic Acids TSDissolve 15 g of met- kaline.
aphosphoric acid in 40 mL of glacial acetic acid and suffi- Neutral Red TSDissolve 100 mg of neutral red in
cient water to make 500 mL. Store in a cold place, and use 100 mL of 50% alcohol.
within 2 days. Nickel Standard Solution TSDissolve 4.78 g of
Methoxyphenylacetic TSDissolve 2.7 g of methoxy- nickel (II) sulfate heptahydrate in water, and dilute with
phenylacetic acid in 6 mL of Tetramethylammonium Hydrox- water to 1000 mL. Immediately prior to use, dilute 10.0 mL
ide TS, and add 20 mL of dehydrated alcohol. Store in a of the solution so obtained with water to 1000 mL. Suitable
polyethylene container. nickel standard solutions are also available commercially.
Methyl Orange TSDissolve 100 mg of methyl orange Ninhydrin TSUse Triketohydrindene Hydrate TS.
in 100 mL of water, and filter if necessary. p-Nitroaniline TSTo 350 mg of p-nitroaniline add
Methyl Purple TSUse Methyl RedMethylene Blue TS. 1.5 mL of hydrochloric acid, and mix. Dilute with water to
Methyl Red TSDissolve 100 mg of methyl red in 50 mL, mix, and allow to settle. Place 5 mL of the clear
100 mL of alcohol, and filter if necessary. supernatant in a 100-mL volumetric flask, and immerse it in
an ice bath. While it is in the ice bath, add 1 mL of hydro-
chloric acid, then add, in small portions, 2 mL of sodium
nitrite solution (1 in 100), dilute with water to volume, and Potassium Acetate TSDissolve 10 g of potassium ace-
mix. tate in water to make 100 mL.
Nitrophenanthroline TSDissolve 150 mg of 5-nitro- PotassiumBismuth Iodide TSDissolve 12.5 g of tar-
1,10-phenanthroline in 15 mL of freshly prepared ferrous taric acid in 25 mL of water, then dissolve 1.06 g of bismuth
sulfate solution (1 in 140). subnitrate in this mixture (Solution A). Dissolve 20 g of po-
Oracet Blue B TSA 1 in 200 solution of oracet blue B tassium iodide in 25 mL of water (Solution B). Dissolve 100 g
in glacial acetic acid. of tartaric acid in 450 mL of water (Solution C). Add Solu-
Orthophenanthroline TSDissolve 150 mg of tions A and B to Solution C, and mix.
orthophenanthroline in 10 mL of a solution of ferrous sul- Potassium Carbonate TSDissolve 7 g of anhydrous po-
fate, prepared by dissolving 700 mg of clear crystals of fer- tassium carbonate in water to make 100 mL.
rous sulfate in 100 mL of water. The ferrous sulfate solution Potassium Chromate TSDissolve 10 g of potassium
must be prepared immediately before dissolving the chromate in water to make 100 mL.
orthophenanthroline. Store in well-closed containers. Potassium Dichromate TSDissolve 7.5 g of potassium
Oxalic Acid TSDissolve 6.3 g of oxalic acid in water to dichromate in water to make 100 mL.
make 100 mL. Potassium Ferricyanide TSDissolve 1 g of potassium
Palladium Chloride TS, BufferedWeigh 500 mg of pal- ferricyanide in 10 mL of water. Prepare this solution fresh.
ladium chloride into a 250-mL beaker, add 5 mL of concen- Potassium Ferrocyanide TSDissolve 1 g of potassium
trated hydrochloric acid, and warm the mixture on a steam ferrocyanide in 10 mL of water. Prepare this solution fresh.
bath. Add 200 mL of hot water in small increments with Potassium Hydroxide TSDissolve 6.5 g of potassium
continued heating until solution is complete. Transfer the hydroxide in water to make 100 mL.
solution to a 250-mL volumetric flask, and dilute with water Potassium Hydroxide TS, AlcoholicUse 0.5 N Potas-
to volume. Transfer 50 mL to a 100-mL volumetric flask. sium Hydroxide, Alcoholic (see in the section Volumetric Solu-
Add 10 mL of 1 M sodium acetate and 9.6 mL of 1 N hydro- tions).
chloric acid. Dilute with water to volume. Potassium Hydroxide TS 2, AlcoholicDissolve 130 g of
Perchloric Acid TSDilute 8.5 mL of perchloric acid with potassium hydroxide, with cooling, in 200 mL of water. Add
water to 100 mL. alcohol to 1000 mL. Store in a well-stoppered dark glass
Phenol TSDissolve 1.2 g of phenol in alcohol to make bottle.
10 mL. Prepare weekly. Potassium Iodide TSDissolve 16.5 g of potassium io-
Phenol Red TS (Phenolsulfonphthalein TS)Dissolve dide in water to make 100 mL. Store in light-resistant con-
100 mg of phenolsulfonphthalein in 100 mL of alcohol, and tainers.
filter if necessary. Potassium Iodide and Starch TSDissolve 0.75 g of po-
pH 4.7 Phenol Red TSDissolve 33 mg of phenolsulfon- tassium iodide in 100 mL of water. Heat to boiling, and
phthalein in 1.5 mL of 2 N sodium hydroxide solution, di- add, with stirring, a solution of 0.5 g of soluble starch in
lute with water to 100 mL, and mix (Solution A). Dissolve 35 mL of water. Boil for 2 minutes, and allow to cool.
25 mg of ammonium sulfate in 235 mL of water, add
SensitivityMix 15 mL in 0.05 mL of glacial acetic acid
105 mL of 2 N sodium hydroxide solution and 135 mL of
and 0.3 mL of diluted iodine TS: a blue color is produced.
2 N acetic acid, and mix (Solution B). Add 25 mL of Solution
Potassium Iodoplatinate TSDissolve 200 mg of pla-
A to Solution B, and mix. If necessary, adjust the pH of this
tinic chloride in 2 mL of water, mix with 25 mL of potas-
solution to 4.7.
sium iodide solution (1 in 25), and add water to make
Phenoldisulfonic Acid TSDissolve 2.5 g of phenol in
50 mL.
15 mL of sulfuric acid in a flask of suitable capacity. Add
Potassium Permanganate TSUse 0.1 N Potassium Per-
7.5 mL of fuming sulfuric acid, stir well, and heat at 100
manganate (see in the section Volumetric Solutions).
for 2 hours. Transfer the product, while still fluid, to a glass-
Potassium Pyroantimonate TSDissolve 2 g of potas-
stoppered bottle, and, when desired for use, warm in a
sium pyroantimonate in 85 mL of hot water. Cool quickly,
water bath until liquefied.
and add 50 mL of a solution containing 50 mg/mL of potas-
Phenolphthalein TSDissolve 1 g of phenolphthalein in
sium hydroxide in water and 1 mL of sodium hydroxide so-
100 mL of alcohol.
lution (8.5 in 100). Allow to stand for 24 h, filter, and dilute
Phenylhydrazine Acetate TSDissolve 10 mL of phenyl-
with water to 150 mL.
hydrazine and 5 mL of glacial acetic acid in water to make
Potassium Sulfate TSDissolve 1 g of potassium sulfate
100 mL.
in water to make 100 mL.
PhenylhydrazineSulfuric Acid TSDissolve 65 mg of
Potassium Thiocyanate TSDissolve 9.7 g of potassium
phenylhydrazine hydrochloride in 100 mL of a cooled mix-
thiocyanate in water to make 100 mL.
ture of equal volumes of sulfuric acid and water.
PyridinePyrazolone TSTo 100 mL of a saturated solu-
Phloroglucinol TSDissolve 500 mg of phloroglucinol in
tion of 1-phenyl-3-methyl-2-pyrazoline-5-one add 20 mL of
25 mL of alcohol. Store in tight containers, protected from
a 1 in 1000 solution of 3,3-dimethyl-1,1-diphenyl-[4,4-bi-
light.
2-pyrazoline]-5,5-dione in pyridine. Store in a dark bottle,
Phosphatic Enzyme TSDissolve 5 g of phosphatic en-
and use within 3 days.
zyme in water to make 50 mL. Prepare this solution fresh.
Pyrogallol TS, AlkalineDissolve 500 mg of pyrogallol
Phosphomolybdic Acid TSDissolve 20 g of phospho-
in 2 mL of water. Dissolve 12 g of potassium hydroxide in
molybdic acid in alcohol to make 100 mL. Filter the solu-
8 mL of water. The solutions should be freshly prepared and
tion, and use only the clear filtrate.
mixed immediately before use.
Phosphotungstic Acid TSDissolve 1 g of phosphotung-
Quinaldine Red TSDissolve 100 mg of quinaldine red
stic acid in water to make 100 mL.
in 100 mL of alcohol.
Picrate TS, AlkalineMix 20 mL of trinitrophenol solu-
Quinone TSDissolve 500 mg of p-benzoquinone in
tion (1 in 100) with 10 mL of sodium hydroxide solution (1
2.5 mL of glacial acetic acid, and dilute with alcohol to
in 20), dilute with water to 100 mL, and mix. Use within 2
50 mL. Prepare this solution fresh daily.
days.
Resorcinol TSDissolve 1 g of resorcinol in hydrochloric
Picric Acid TSSee Trinitrophenol TS.
acid to make 100 mL.
Platinic Chloride TSDissolve 2.6 g of platinic chloride
Ruthenium Red TSDissolve 10 g of lead acetate in
in water to make 20 mL.
water, dilute with water to 100 mL, and add 80 mg of ru-
PlatinumCobalt TSDissolve 1.246 g of potassium
thenium red. The solution is wine-red in color. [NOTEIf
chloroplatinate (K2PtCl6) and 1.000 g of cobalt chloride
necessary, add additional ruthenium red to obtain a wine-
(CoCl2 6H2O) in water, add 100 mL of hydrochloric acid,
red color.]
and dilute with water to 1 L.
Saline TSDissolve 9.0 g of sodium chloride in water to Sodium Hypochlorite TSUse Sodium Hypochlorite Solu-
make 1000 mL. tion (see in the section Reagent Specifications).
[NOTEWhere pyrogen-free saline TS is specified in this Sodium Iodohydroxyquinolinesulfonate TSDissolve
Pharmacopeia, saline TS that has met the requirements of 8.8 g of iodohydroxyquinoline sulfonic acid in 200 mL of
the Pyrogen Test 151 is to be used.] water, and add 6.5 mL of 4 N sodium hydroxide. Dilute
Saline TS, Pyrogen-FreeSee Saline TS. with water to 250 mL, mix, and filter.
Schweitzers ReagentSee Cupric Oxide, Ammoniated, Sodium Nitroferricyanide TSDissolve 1 g of sodium
TS. nitroferricyanide in water to make 20 mL. Prepare this solu-
SilverAmmoniaNitrate TSDissolve 1 g of silver ni- tion fresh.
trate in 20 mL of water. Add ammonia TS, dropwise, with Dibasic Sodium Phosphate TSDissolve 12 g of dibasic
constant stirring, until the precipitate is almost but not en- sodium phosphate in water to make 100 mL.
tirely dissolved. Filter, and store in tight, light-resistant con- Sodium Phosphotungstate TSTo a solution of 20 g of
tainers. sodium tungstate in 100 mL of water add sufficient phos-
SilverAmmonium Nitrate TSSee SilverAmmonia phoric acid to impart a strongly acid reaction to litmus, and
Nitrate TS. filter. When required for use, decant the clear solution from
Silver Diethyldithiocarbamate TSDissolve 1 g of silver any sediment that may be present. Store in tight, light-resis-
diethyldithiocarbamate in 200 mL of pyridine from a freshly tant containers.
opened bottle or that which has been recently distilled. Sodium Sulfide TSDissolve 1 g of sodium sulfide in
Store in light-resistant containers, and use within 30 days. water to make 10 mL. Prepare this solution fresh.
Silver Nitrate TSUse 0.1 N Silver Nitrate (see in the Sodium Tartrate TSDissolve 11.5 g of sodium tartrate
section Volumetric Solutions). in water to make 100 mL.
Simulated Gastric Fluid TSSee Gastric Fluid, Simulated, Sodium Tetraphenylboron TSDissolve 1.2 g of sodium
TS. tetraphenylboron in water to make 200 mL. If necessary, stir
Simulated Intestinal Fluid TSSee Intestinal Fluid, Simu- for 5 minutes with 1 g of aluminum oxide, and filter to clar-
lated, TS. ify.
Sodium Acetate TSDissolve 13.6 g of sodium acetate Sodium Thioglycolate TSDissolve 1.5 g of sodium thi-
in water to make 100 mL. oglycolate in 450 mL of water, and add 50 mL of alcohol.
Sodium Alizarinsulfonate TSDissolve 100 mg of so- Use within 3 days.
dium alizarinsulfonate in 100 mL of water, and filter. Sodium Thiosulfate TSUse 0.1 N Sodium Thiosulfate
Sodium Aminoacetate TS (Sodium Glycinate TS)Dis- (see in the section Volumetric Solutions).
solve 3.75 g of aminoacetic acid in about 500 mL of water, Standard Lead SolutionSee under Heavy Metals 231.
add 2.1 g of sodium hydroxide, and dilute with water to Stannous Chloride, Acid, TSDissolve 8 g of stannous
1000 mL. Mix 9 mL of the resulting solution with 1 mL of chloride in 500 mL of hydrochloric acid. Store in glass con-
dilute glacial acetic acid (1 in 300). This test solution has a tainers, and use within 3 months.
pH between 10.4 and 10.5. Stannous Chloride, Acid, Stronger, TSDissolve 40 g
Sodium Bisulfite TSDissolve 10 g of sodium bisulfite in of stannous chloride in 100 mL of hydrochloric acid. Store in
water to make 30 mL. Prepare this solution fresh. glass containers, and use within 3 months.
Sodium Bitartrate TSDissolve 1 g of sodium bitartrate Starch TSMix 1 g of soluble starch with 10 mg of red
in water to make 10 mL. Prepare this solution fresh. mercuric iodide and sufficient cold water to make a thin
Sodium Carbonate TSDissolve 10.6 g of anhydrous so- paste. Add 200 mL of boiling water, and boil for 1 minute
dium carbonate in water to make 100 mL. with continuous stirring. Cool, and use only the clear solu-
Sodium Chloride TS, AlkalineDissolve 2 g of sodium tion. [NOTECommercially available, stabilized starch indica-
hydroxide in 100 mL of water, saturate the solution with tor solutions may be used, including mercury-free solutions
sodium chloride, and filter. preserved with other compounds such as salicylic acid.]
Sodium Citrate TSDissolve 73.5 g of sodium citrate di- Starch, Iodide-Free, TSMix 1 g of soluble starch with
hydrate in water to make 250 mL. sufficient cold water to make a thin paste. While stirring,
Sodium Citrate TS, AlkalineDissolve 50 g of sodium add 100 mL of boiling water, and allow to cool. Prepare this
citrate dihydrate and 2.5 g of sodium hydroxide in water to solution immediately before use. Iodide-free starch TS shows
make 250 mL. a blue color when 20 mL of potassium iodide solution (1 in
Sodium Cobaltinitrite TSDissolve 10 g of sodium 400) and 0.05 mL of an iodinepotassium iodide solution
cobaltinitrite in water to make 50 mL, and filter if necessary. (prepared by dissolving 127 mg of iodine and 800 mg of
Sodium Fluoride TSDry about 500 mg of sodium fluo- potassium iodide in water and diluting with water to
ride at 200 for 4 hours. Accurately weigh 222 mg of the 100 mL) are added to 1 mL of the iodide-free starch TS.
dried material, and dissolve in water to make 100.0 mL. Pi- Starch Iodide Paste TSHeat 100 mL of water in a
pet 10 mL of this solution into a 1-L volumetric flask, and 250-mL beaker to boiling, add a solution of 0.75 g of potas-
dilute with water to volume. Each mL of this solution corre- sium iodide in 5 mL of water, then add 2 g of zinc chloride
sponds to 0.01 mg of fluorine (F). dissolved in 10 mL of water, and, while the solution is boil-
Sodium Hydrosulfite TS, AlkalineDissolve 25 g of po- ing, add, with stirring, a smooth suspension of 5 g of solu-
tassium hydroxide in 35 mL of water, and 50 g of sodium ble starch in 30 mL of cold water. Continue to boil for
hydrosulfite in 250 mL of water. When the test solution is 2 minutes, then cool. Store in well-closed containers in a
required, mix 40 mL of the hydroxide solution with the cold place.
250 mL of the hydrosulfite solution. Prepare this solution Starch iodide paste TS must show a definite blue streak
fresh. when a glass rod, dipped in a mixture of 1 mL of 0.1 M
Sodium Hydroxide TSDissolve 4.0 g of sodium hy- sodium nitrite, 500 mL of water, and 10 mL of hydrochloric
droxide in water to make 100 mL. acid, is streaked on a smear of the paste.
Sodium Hydroxide TS 2Transfer 8.5 g of sodium hy- StarchPotassium Iodide TSDissolve 500 mg of potas-
droxide to a 100-mL volumetric flask, and dissolve in and sium iodide in 100 mL of freshly prepared starch TS. Prepare
dilute with water to volume. this solution fresh.
Sodium Hydroxide TS 3Prepare a 420-mg/mL solution Stronger Cupric Acetate TSSee Cupric Acetate TS,
of sodium hydroxide in water. Stronger.
Sodium Hypobromite TSTo a solution of 20 g of so- Sudan III TSDissolve 0.05 g of Sudan III in 25 mL of
dium hydroxide in 75 mL of water add 5 mL of bromine. alcohol, with warming if necessary. Cool, add 25 mL of
After solution has taken place, dilute with water to 100 mL. glycerin, and mix. Filter if undissolved material persists.
Prepare this solution fresh. Sudan IV TSDissolve 0.5 g of Sudan IV in chloroform
to make 100 mL.
Sulfanilic Acid TSDissolve 800 mg of sulfanilic acid in Triphenyltetrazolium Chloride TSDissolve 500 mg of
100 mL of acetic acid. Store in tight containers. triphenyltetrazolium chloride in dehydrated alcohol to make
Diazotized Sulfanilic Acid TSDissolve 0.9 g of sulfanilic 100 mL.
acid in 9 mL of hydrochloric acid with warming, and dilute Xylenol Orange TSDissolve 100 mg of xylenol orange
with water to 100 mL. Cool 10 mL of this solution in iced in 100 mL of alcohol.
water, and add 10 mL of a sodium nitrite solution (4.5 in Zinc Uranyl Acetate TSDissolve 50 g of uranyl acetate
100) previously cooled in iced water. Allow to stand at 0 in a mixture of 15 mL of glacial acetic acid and water to
for at least 15 minutes (the solution may be kept for 3 days make 500 mL. Then dissolve 150 g of zinc acetate in a mix-
at this temperature). Immediately before use, add 20 mL of ture of 15 mL of glacial acetic acid and water to make
sodium carbonate solution (1 in 10). 500 mL. Mix the two solutions, allow to stand overnight,
Sulfanilic-1-Naphthylamine TSDissolve 500 mg of sul- and pass through a dry filter, if necessary.
fanilic acid in 150 mL of acetic acid. Dissolve 100 mg of
1-naphthylamine hydrochloride in 150 mL of acetic acid,
VOLUMETRIC SOLUTIONS
and mix the two solutions. The pink color that may develop
on standing can be removed by treatment with zinc.
Normal SolutionsNormal solutions are solutions that
Sulfanilic--Naphthylamine TSSee Sulfanilic-1-Naph-
contain 1 gram equivalent weight of the active substance in
thylamine TS.
each 1000 mL of solution; that is, an amount equivalent to
Sulfomolybdic Acid TSDissolve, with the aid of heat,
1.0079 g of hydrogen or 7.9997 g of oxygen. Normal solu-
2.5 g of ammonium molybdate in 20 mL of water, add
tions and solutions bearing a specific relationship to normal
50 mL of 12 N sulfuric acid, and dilute with water to
solutions, and used in volumetric determinations, are desig-
100 mL. Store this solution in a polyethylene container.
nated as follows: normal, 1 N; double-normal, 2 N; half-nor-
Sulfuric Acid TSAdd a quantity of sulfuric acid of
mal, 0.5 N; tenth-normal, 0.1 N; fiftieth-normal, 0.02 N;
known concentration to sufficient water to adjust the final
hundredth-normal, 0.01 N; thousandth-normal, 0.001 N.
concentration to between 94.5% and 95.5% (w/w) of
Molar SolutionsMolar solutions are solutions that con-
H2SO4.
tain, in 1000 mL, 1 gram-molecule of the reagent. Thus,
[NOTESince the acid concentration may change upon
each liter of a molar solution of sulfuric acid contains
standing or upon intermittent use, the concentration should
98.07 g of H2SO4 and each liter of a molar solution of potas-
be checked frequently and solutions assaying more than
sium ferricyanide contains 329.25 g of K3Fe(CN)6. Solutions
95.5% or less than 94.5% discarded.]
containing, in 1000 mL, one-tenth of a gram-molecule of
Sulfuric AcidFormaldehyde TSAdd 1 drop of formal-
the reagent are designated tenth-molar, 0.1 M; and other
dehyde TS to each mL of sulfuric acid, and mix. Prepare this
molarities are similarly indicated.
solution fresh.
Empirical SolutionsIt is frequently difficult to prepare
Tannic Acid TSDissolve 1 g of tannic acid in 1 mL of
standard solutions of a desired theoretical normality, and
alcohol, and dilute with water to 10 mL. Prepare this solu-
this is not essential. A solution of approximately the desired
tion fresh.
normality is prepared and standardized by titration against a
Tartaric Acid TSDissolve 3 g of tartaric acid in water to
primary standard solution. The normality factor so obtained
make 10 mL. Prepare this solution fresh.
is used in all calculations where such empirical solutions are
Tetrabromophenolphthalein Ethyl Ester TSDissolve
employed. If desired, an empirically prepared solution may
100 mg of tetrabromophenolphthalein ethyl ester in 90 mL
be adjusted downward to a given normality provided it is
of glacial acetic acid, and dilute with glacial acetic acid to
strong enough to permit dilution.
100 mL. Prepare this solution fresh.
All volumetric solutions, whether made by direct solution
Tetramethylammonium Hydroxide TSUse an aqueous
or by dilution of a stronger solution, must be thoroughly
solution containing, in each 100 mL, the equivalent of 10 g
mixed by shaking before standardization. As the strength of
of anhydrous tetramethylammonium hydroxide.
a standard solution may change upon standing, the factor
Thioacetamide TSDissolve 4 g of thioacetamide in
should be redetermined frequently.
100 mL of water.
When solutions of a reagent are used in several normali-
ThioacetamideGlycerin Base TSMix 0.2 mL of thio-
ties, the details of the preparation and standardization are
acetamide TS and 1 mL of glycerin base TS, and heat in a
usually given for the normality most frequently required.
boiling water bath for 20 seconds. Use the mixture immedi-
Stronger or weaker solutions are prepared and standardized
ately.
in the same general manner as described, using proportion-
Thorium Nitrate TSDissolve 1 g of thorium nitrate in
ate amounts of the reagent. It is possible in many instances
water to make 100 mL. Filter, if necessary.
to prepare lower normalities accurately by making an exact
Thymol Blue TSDissolve 100 mg of thymol blue in
dilution of a stronger solution. Volumetric solutions prepared
100 mL of alcohol, and filter if necessary.
by dilution should be restandardized either as directed for
Thymolphthalein TSDissolve 100 mg of thymolphtha-
the stronger solution or by comparison with another volu-
lein in 100 mL of alcohol, and filter if necessary.
metric solution having a known ratio to the stronger
Titanium Trichloride TSDissolve 15 g of titanium
solution.
trichloride in 100 mL of 10% hydrochloric acid solution.
Dilute solutions that are not stable, as, for instance, potas-
Titanium TrichlorideSulfuric Acid TSMix carefully
sium permanganate 0.01 N and more dilute sodium thiosul-
20 mL of titanium trichloride TS in 13 mL of sulfuric acid.
fate, are preferably prepared by exactly diluting the higher
Add sufficient 30% hydrogen peroxide to produce a yellow
normality with thoroughly boiled and cooled water on the
color. Heat until white fumes are evolved, allow to cool, and
same day they are required for use.
dilute with water. Repeat the evaporation and addition of
Blank DeterminationsWhere it is directed that any
water until a colorless solution is obtained. Dilute with water
necessary correction be made by a blank determination,
to 100 mL.
the determination is to be conducted with the use of the
p-Toluenesulfonic Acid TSDissolve 2 g of p-tolu-
same quantities of the same reagents treated in the same
enesulfonic acid in 10 mL of a mixture of 7 parts of acetone
manner as the solution or mixture containing the portion of
and 3 parts of water.
the substance under assay or test, but with the substance
Triketohydrindene Hydrate TS (Ninhydrin TS)Dissolve
itself omitted. Appropriate blank corrections are to be made
200 mg of triketohydrindene hydrate in water to make
for all Pharmacopeial titrimetric assays (see Titrimetry 541).
10 mL. Prepare this solution fresh.
All Pharmacopeial assays that are volumetric in nature in-
Trinitrophenol TS (Picric Acid TS)Dissolve the equiva-
dicate the weight of the substance being assayed to which
lent of 1 g of anhydrous trinitrophenol in 100 mL of hot
each mL of the primary volumetric solution is equivalent. In
water. Cool the solution, and filter if necessary.
general, these equivalents may be derived by simple calcula-
USP 36 Solutions / Volumetric Solutions 1219
tion from the data given under Molecular Formulas and Bromine, Tenth-Normal (0.1 N)
Weights, in the Reference Tables. Br, 79.90
7.990 g in 1000 mL
Dissolve 3 g of potassium bromate and 15 g of potassium
Preparation and Methods of Standardization
bromide in water to make 1000 mL, and standardize the
of Volumetric Solutions solution as follows.
Accurately measure about 25 mL of the solution into a
The following directions give only one method for stan- 500-mL iodine flask, and dilute with 120 mL of water. Add
dardization, but other methods of standardization, capable 5 mL of hydrochloric acid, insert the stopper in the flask,
of yielding at least the same degree of accuracy, may be and shake it gently. Then add 5 mL of potassium iodide TS,
used. The values obtained in the standardization of volumet- again insert the stopper, shake the mixture, allow it to stand
ric solutions are valid for all Pharmacopeial uses of these for 5 minutes, and titrate the liberated iodine with 0.1 N
solutions, regardless of the instrumental or chemical indica- sodium thiosulfate VS, adding 3 mL of starch TS as the
tors employed in the individual monographs. Where the ap- endpoint is approached.
parent normality or molarity of a titrant depends upon the Preserve in dark amber-colored, glass-stoppered bottles.
special conditions of its use, the individual monograph sets
forth the directions for standardizing the reagent in the
specified context. For those salts that usually are available as
certified primary standards, or that are available as highly
purified salts of primary standard quality, it is permissible to
prepare solutions by accurately weighing a suitable quantity
of the salt and dissolving it to produce a specific volume of
Ceric Ammonium Nitrate, Twentieth-Normal (0.05 N)
solution of known concentration. Acetic, hydrochloric, and
Ce(NO3)4 2NH4NO3, 548.22
sulfuric acids may be standardized against a sodium hydrox-
2.741 g in 100 mL
ide solution that recently has been standardized against a
Dissolve 2.75 g of ceric ammonium nitrate in 1 N nitric
certified primary standard.
acid to obtain 100 mL of solution, and filter. Standardize the
All volumetric solutions, if practicable, are to be prepared,
solution as follows.
standardized, and used at the standard temperature of 25.
Accurately measure 10 mL of freshly standardized 0.1 N
If a titration is carried out with the volumetric solution at a
ferrous ammonium sulfate VS into a flask, and dilute with
markedly different temperature, standardize the volumetric
water to about 100 mL. Add 1 drop of nitrophenanthroline
solution used as the titrant at that different temperature, or
TS, and titrate with the ceric ammonium nitrate solution to
make a suitable temperature correction.
a colorless endpoint.
Acetic Acid, Double-Normal (2 N)
C2H4O2, 60.05
120.10 g in 1000 mL
Add 116 mL of glacial acetic acid to sufficient water to
make 1000 mL after cooling to room temperature.
Ceric Sulfate, Tenth-Normal (0.1 N)
Ammonium Thiocyanate, Tenth-Normal (0.1 N)
Ce(SO4)2, 332.24
NH4SCN, 76.12
33.22 g in 1000 mL
7.612 g in 1000 mL
Use commercially available volumetric standard solution.
Dissolve about 8 g of ammonium thiocyanate in 1000 mL
Standardize the solution as follows.
of water, and standardize the solution as follows.
Accurately weigh about 0.2 g of sodium oxalate, primary
Accurately measure about 30 mL of 0.1 N silver nitrate VS
standard, dried according to the instructions on its label,
into a glass-stoppered flask. Dilute with 50 mL of water,
and dissolve in 75 mL of water. Add, with stirring, 2 mL of
then add 2 mL of nitric acid and 2 mL of ferric ammonium
sulfuric acid that has previously been mixed with 5 mL of
sulfate TS, and titrate with the ammonium thiocyanate solu-
water, mix well, add 10 mL of hydrochloric acid, and heat
tion to the first appearance of a red-brown color.
to between 70 and 75. Titrate with 0.1 N ceric sulfate to a
permanent slight yellow color. Each 6.700 mg of sodium ox-
alate is equivalent to 1 mL of 0.1 N ceric sulfate.
If desirable, 0.1 N ammonium thiocyanate may be re-
placed by 0.1 N potassium thiocyanate where the former is
directed in various tests and assays.
Cupric Nitrate, Tenth-Normal (0.1 N)
Bismuth Nitrate, 0.01 M
Cu(NO3)2 2.5H2O, 232.59
Bi(NO3)3 5H2O, 485.07
23.26 g in 1000 mL
1000 mL of this solution contains 4.851 g of bismuth nitrate
Cu(NO3)2 3H2O, 241.60
pentahydrate
24.16 g in 1000 mL
Dissolve 4.86 g of bismuth nitrate pentahydrate in 60 mL
Dissolve 23.3 g of cupric nitrate 2.5 hydrate, or 24.2 g of
of dilute nitric acid, add 0.01 N nitric acid to make
the trihydrate, in water to make 1000 mL. Standardize the
1000 mL, and standardize the solution as follows.
Accurately measure 25 mL of the prepared bismuth nitrate
Transfer 20.0 mL of the solution to a 250-mL beaker. Add
solution, add 50 mL of water and 1 drop of xylenol orange
2 mL of 5 M sodium nitrate, 20 mL of ammonium acetate
TS, and titrate the solution with 0.01 M edetate disodium
TS, and sufficient water to make 100 mL. Titrate with
VS until the red color changes to yellow. Calculate the mo-
0.05 M edetate disodium VS. Determine the endpoint po-
larity factor.
tentiometrically using a cupric ion-double junction reference
1220 Volumetric Solutions / Solutions USP 36
electrode system. Perform a blank determination, and make Store in tight containers, protected from light.
any necessary correction.
Standard DichlorophenolIndophenol Solution Ferrous Ammonium Sulfate, Tenth-Normal (0.1 N)
To 50 mg of 2,6-dichlorophenolindophenol sodium that Fe(NH4)2(SO4)2 6H2O, 392.14
has been stored in a desiccator over soda lime add 50 mL of 39.21 g in 1000 mL
water containing 42 mg of sodium bicarbonate, shake vigor- Dissolve 40 g of ferrous ammonium sulfate in a previously
ously, and when the dye is dissolved, add water to make cooled mixture of 40 mL of sulfuric acid and 200 mL of
200 mL. Filter into an amber, glass-stoppered bottle. Use water, dilute with water to 1000 mL, and mix. On the day
within 3 days and standardize immediately before use. Stan- of use, standardize the solution as follows.
dardize the solution as follows. Accurately measure 25 to 30 mL of the solution into a
Accurately weigh 50 mg of USP Ascorbic Acid RS, and flask, add 2 drops of orthophenanthroline TS, and titrate
transfer to a glass-stoppered, 50-mL volumetric flask with with 0.1 N ceric sulfate VS until the red color is changed to
the aid of a sufficient volume of metaphosphoricacetic pale blue.
acids TS to make 50 mL. Immediately transfer 2 mL of the
ascorbic acid solution to a 50-mL conical flask containing
5 mL of the metaphosphoricacetic acids TS, and titrate rap-
idly with the dichlorophenolindophenol solution until a dis-
tinct rose-pink color persists for at least 5 seconds. Perform a
blank titration by titrating 7 mL of the metaphosphoric
acetic acids TS plus a volume of water equal to the volume
Hydrochloric Acid, Alcoholic, Tenth-Molar (0.1 M)
of the dichlorophenol solution used in titrating the ascorbic
HCl, 36.46
acid solution. Express the concentration of the standard so-
Dilute 9.0 mL of hydrochloric acid to 1000 mL with al-
lution in terms of its equivalent in mg of ascorbic acid.
dehydefree alcohol.
Edetate Disodium, Twentieth-Molar (0.05 M)
Hydrochloric Acid, Half-Normal (0.5 N)
C10H14N2Na2O8 2H2O, 372.24
HCl, 36.46
18.61 g in 1000 mL
18.23 g in 1000 mL
Dissolve 18.6 g of edetate disodium in water to make
To a 1000-mL volumetric flask containing 40 mL of water
slowly add 43 mL of hydrochloric acid. Cool, and add water
Accurately weigh about 200 mg of chelometric standard
to volume. Standardize the solution as follows.
calcium carbonate, previously dried at 110 for 2 hours and
Accurately weigh about 2.5 g of tromethamine, dried ac-
cooled in a desiccator, or dried according to the label in-
cording to the label instructions or, if this information is not
structions, transfer to a 400-mL beaker, add 10 mL of water,
available, dried at 105 for 3 h. Dissolve in 50 mL of water,
and swirl to form a slurry. Cover the beaker with a watch
and add 2 drops of bromocresol green TS. Titrate with 0.5
glass, and introduce 2 mL of diluted hydrochloric acid from
N hydrochloric acid to a pale yellow endpoint. Each
a pipet inserted between the lip of the beaker and the edge
60.57 mg of tromethamine is equivalent to 1 mL of 0.5 N
of the watch glass. Swirl the contents of the beaker to dis-
hydrochloric acid.
solve the calcium carbonate. Wash down the sides of the
beaker, the outer surface of the pipet, and the watch glass
with water, and dilute with water to about 100 mL. While
stirring the solution, preferably with a magnetic stirrer, add
about 30 mL of the edetate disodium solution from a 50-mL
buret. Add 15 mL of sodium hydroxide TS and 300 mg of
hydroxy naphthol blue, and continue the titration with the
Hydrochloric Acid, Half-Normal (0.5 N) in Methanol
edetate disodium solution to a blue endpoint.
HCl, 36.46
18.23 g in 1000 mL
To a 1000-mL volumetric flask containing 40 mL of water
slowly add 43 mL of hydrochloric acid. Cool, and add meth-
anol to volume. Standardize the solution as follows.
Ferric Ammonium Sulfate, Tenth-Normal (0.1 N)
available, dried at 105 for 3 h. Proceed as directed under
FeNH4(SO4)2 12H2O, 482.19
Hydrochloric Acid, Normal (1 N), beginning with Dissolve in
48.22 g in 1000 mL
50 mL of water.
Dissolve 50 g of ferric ammonium sulfate in a mixture of
300 mL of water and 6 mL of sulfuric acid, dilute with water
to 1000 mL, and mix. Standardize the solution as follows.
Accurately measure about 40 mL of the solution into a
glass-stoppered flask, add 5 mL of hydrochloric acid, mix,
and add a solution of 3 g of potassium iodide in 10 mL of
water. Insert the stopper, allow to stand for 10 minutes,
Hydrochloric Acid, Normal (1 N)
then titrate the liberated iodine with 0.1 N sodium thiosul-
HCl, 36.46
fate VS, adding 3 mL of starch TS as the endpoint is ap-
36.46 g in 1000 mL
proached. Correct for a blank run on the same quantities of
Dilute 85 mL of hydrochloric acid with water to 1000 mL.
the same reagents.
available, dried at 105 for 3 h. Dissolve in 50 mL of water,
and add 2 drops of bromocresol green TS. Titrate with 1 N
hydrochloric acid to a pale yellow endpoint. Each 0.1 M Lead NitrateDissolve 33 g of lead nitrate in
121.14 mg of tromethamine is equivalent to 1 mL of 1 N 1000 mL of water. Standardize the solution as follows. To
hydrochloric acid. 20.0 mL of the lead nitrate solution add 300 mL of water.
Add about 50 mg of Xylenol Orange Triturate, and add me-
thenamine until the solution becomes violet-pink. Titrate
with 0.1 M edetate disodium VS to the yellow endpoint.
Calculate the molarity.
Dilute 50.0 mL of 0.1 M Lead Nitrate to 500.0 mL with
water.
Iodine, Tenth-Normal (0.1 N)
I, 126.90 Lead Perchlorate, Tenth-Molar (0.1 M)
12.69 g in 1000 mL Pb(ClO4)2 3H2O, 460.15
Dissolve about 14 g of iodine in a solution of 36 g of po- 46.01 g in 1000 mL
tassium iodide in 100 mL of water, add 3 drops of hydro- Dissolve 46 g of lead perchlorate in water, and dilute with
chloric acid, dilute with water to 1000 mL, and standardize water to 1000.0 mL. Accurately weigh about 150 mg of so-
the solution as follows. dium sulfate, previously dried at 105 for 4 hours, and dis-
Transfer 25.0 mL of the iodine solution to a 250-mL flask, solve in 50 mL of water. Add 50 mL of a mixture of water
dilute with water to 100 mL, add 1 mL of 1 N hydrochloric and formaldehyde (1:1), and stir for about 1 minute. Deter-
acid, swirl gently to mix, and titrate with 0.1 N sodium mine the endpoint potentiometrically using a lead ion selec-
thiosulfate VS until the solution has a pale yellow color. Add tive electrode. Perform a blank determination, and make
2 mL of starch TS and continue titrating until the solution is any necessary corrections. Each 14.204 mg of sodium sulfate
colorless. is equivalent to 1 mL of 0.1 M lead perchlorate.
Preserve in amber-colored, glass-stoppered bottles.
Lead Perchlorate, Hundredth-Molar (0.01 M)
Iodine, Twentieth-Normal (0.05 N) Pb(ClO4)2, 406.10
I, 126.90 Accurately pipet 100 mL of commercially available 0.1 M
6.33 g in 1000 mL lead perchlorate solution into a 1000-mL volumetric flask,
Dissolve about 6.5 g of iodine in a solution of 18 g of add a sufficient quantity of water to make 1000 mL, and
potassium iodide in 100 mL of water, add 3 drops of hydro- standardize the solution as follows.
chloric acid, dilute with water to 1000 mL, and standardize Accurately pipet 50 mL of 0.01 M lead perchlorate solu-
the solution as follows. tion, as prepared above, into a 250-mL conical flask. Add
Transfer 50.0 mL of the iodine solution to a 250-mL flask, 3 mL of aqueous hexamethylenetetramine solution (2.0 g
dilute with water to 100 mL, add 1 mL of 1 N hydrochloric per 100 mL) and 4 drops of 0.5% xylenol orange indicator
acid, swirl gently to mix, and titrate with 0.1 N sodium prepared by adding 500 mg of xylenol orange to 10 mL of
thiosulfate VS until the solution has a pale yellow color. Add alcohol and diluting with water to 100 mL. (Omit the alco-
2 mL of starch TS, and continue titrating until the solution is hol if the sodium salt of the indicator is used). Titrate with
colorless. 0.05 M edetate disodium VS to a yellow endpoint.
Iodine, Hundredth-Normal (0.01 N) Lithium Methoxide, Fiftieth-Normal (0.02 N) in
I, 126.90 Methanol
1.269 g in 1000 mL CH3LiO, 37.97
Dissolve about 1.4 g of iodine in a solution of 3.6 g of 759.6 mg in 1000 mL
potassium iodide in 100 mL of water, add 3 drops of hydro- Dissolve 0.12 g of freshly cut lithium metal in 150 mL of
chloric acid, dilute with water to 1000 mL, and standardize methanol, cooling the flask during addition of the metal.
the solution as follows. When the reaction is complete, add 850 mL of methanol,
Transfer 100.0 mL of iodine solution to a 250-mL flask, and mix. Store the solution preferably in the reservoir of an
add 1 mL of 1 N hydrochloric acid, swirl gently to mix, and automatic delivery buret suitably protected from carbon di-
titrate with 0.1 N sodium thiosulfate VS until the solution oxide and moisture. Standardize the solution by titration
has a pale yellow color. Add 2 mL of starch TS, and con- against benzoic acid as described under Sodium Methoxide,
tinue titrating until the solution is colorless. Tenth-Normal (0.1 N) (in Toluene), but use only 100 mg of
Preserve in amber-colored, glass-stoppered bottles. benzoic acid. Each 2.442 mg of benzoic acid is equivalent to
1 mL of 0.02 N lithium methoxide.
[NOTERestandardize the solution frequently.]
Lead Nitrate, Hundredth-Molar (0.01 M)
Pb (NO3)2, 331.21
Lithium Methoxide, Tenth-Normal (0.1 N) in
3.312 g in 1000 mL
Chlorobenzene
Xylenol Orange TriturateTriturate 1 part of xylenol or-
CH3OLi, 37.97
ange with 99 parts of potassium nitrate.
3.798 g in 1000 mL
Dissolve 700 mg of freshly cut lithium metal in 150 mL of Dissolve about 35 g of mercuric nitrate in a mixture of
methanol, cooling the flask during addition of the metal. 5 mL of nitric acid and 500 mL of water, and dilute with
When the reaction is complete, add 850 mL of chloroben- water to 1000 mL. Standardize the solution as follows.
zene. If cloudiness or precipitation occurs, add sufficient Transfer an accurately measured volume of about 20 mL
methanol to clarify the solution. Store preferably in the res- of the solution to a conical flask, and add 2 mL of nitric acid
ervoir of an automatic delivery buret suitably protected from and 2 mL of ferric ammonium sulfate TS. Cool to below
carbon dioxide and moisture. Standardize the solution by 20, and titrate with 0.1 N ammonium thiocyanate VS to
titration against benzoic acid as described under Sodium the first appearance of a permanent brownish color.
Methoxide, Tenth-Normal (0.1 N) (in Toluene).
Oxalic Acid, Tenth-Normal (0.1 N)
H2C2O4 2H2O, 126.07
Lithium Methoxide, Tenth-Normal (0.1 N) in Methanol
6.303 g in 1000 mL
CH3OLi, 37.97
Dissolve 6.45 g of oxalic acid in water to make 1000 mL.
3.798 g in 1000 mL
Standardize by titration against freshly standardized 0.1 N
Dissolve 700 mg of freshly cut lithium metal in 150 mL of
potassium permanganate VS as directed under Potassium
methanol, cooling the flask during addition of the metal.
Permanganate, Tenth-Normal (0.1 N).
When the reaction is complete, add 850 mL of methanol. If
Preserve in glass-stoppered bottles, protected from light.
cloudiness or precipitation occurs, add sufficient methanol
to clarify the solution. Store preferably in the reservoir of an
automatic delivery buret suitably protected from carbon di-
oxide and moisture. Standardize the solution by titration
against benzoic acid as described under Sodium Methoxide,
Tenth-Normal (0.1 N) (in Toluene).
Perchloric Acid, Tenth-Normal (0.1 N) in Dioxane
Mix 8.5 mL of perchloric acid with sufficient dioxane to
make 1000 mL. Standardize the solution as follows.
Accurately weigh about 700 mg of potassium biphthalate,
previously crushed lightly and dried at 120 for 2 hours, and
dissolve in 50 mL of glacial acetic acid in a 250-mL flask.
Add 2 drops of crystal violet TS, and titrate with the per- Lithium Methoxide, Tenth-Normal (0.1 N) in Toluene
chloric acid solution until the violet color changes to bluish CH3OLi, 37.97
green. Carry out a blank determination. Each 20.423 mg of 3.798 g in 1000 mL
potassium biphthalate is equivalent to 1 mL of 0.1 N per- Dissolve 700 mg of freshly cut lithium metal in 150 mL of
chloric acid. methanol, cooling the flask during addition of the metal.
When reaction is complete, add 850 mL of toluene. If cloud-
iness or precipitation occurs, add sufficient methanol to clar-
ify the solution. Store preferably in the reservoir of an auto-
matic delivery buret suitably protected from carbon dioxide
and moisture. Standardize the solution by titration against
benzoic acid as described under Sodium Methoxide, Tenth-
Perchloric Acid, Tenth-Normal (0.1 N) in Glacial Acetic
Normal (0.1 N) (in Toluene).
Acid
HClO4, 100.46
10.05 g in 1000 mL
[NOTEWhere called for in the tests and assays, this volu-
metric solution is specified as 0.1 N perchloric acid. Thus,
where 0.1 N or other strength of this volumetric solution is
specified, the solution in glacial acetic acid is to be used,
unless the words in dioxane are stated. [See also Perchloric Magnesium Chloride, 0.01 M
Acid, Tenth-Normal (0.1 N) in Dioxane.]] MgCl2 6H2O, 203.30
Mix 8.5 mL of perchloric acid with 500 mL of glacial ace- 2.0330 g in 1000 mL
tic acid and 21 mL of acetic anhydride, cool, and add glacial Dissolve about 2.04 g of magnesium chloride in 1000 mL
acetic acid to make 1000 mL. Alternatively, the solution may of freshly boiled and cooled water, and standardize the solu-
be prepared as follows. Mix 11 mL of 60 percent perchloric tion as follows.
acid with 500 mL of glacial acetic acid and 30 mL of acetic Accurately measure 25 mL of the prepared magnesium
anhydride, cool, and add glacial acetic acid to make chloride solution. Add 50 mL of water, 3 mL of am-
1000 mL. moniaammonium chloride buffer TS and 0.04 g of eri-
Allow the prepared solution to stand for 1 day for the ochrome black Tsodium chloride reagent. Titrate with
excess acetic anhydride to be combined, and determine the 0.05 M edetate disodium VS until the red-purple color of
water content by Method I (see Water Determination 921), the solution changes to blue-purple.
except to use a test specimen of about 5 g of the 0.1 N
perchloric acid that is expected to contain approximately
1 mg of water and the Reagent (see Reagent under Method
Ia in Water Determination 921) diluted such that 1 mL is
equivalent to about 1 to 2 mg of water. If the water content
exceeds 0.5%, add more acetic anhydride. If the solution
Mercuric Nitrate, Tenth-Molar (0.1 M) contains no titratable water, add sufficient water to obtain a
Hg(NO3)2, 324.60 content of between 0.02% and 0.5% of water. Allow the
32.46 g in 1000 mL solution to stand for 1 day, and again titrate the water con-
tent. The solution so obtained contains between 0.02% and Potassium Ferricyanide, Twentieth-Molar (0.05 M)
0.5% of water, indicating freedom from acetic anhydride. K3Fe(CN)6, 329.24
Standardize the solution as follows. 16.46 g in 1000 mL
Accurately weigh about 700 mg of potassium biphthalate, Dissolve about 17 g of potassium ferricyanide in water to
previously crushed lightly and dried at 120 for 2 hours, and make 1000 mL. Standardize the solution as follows.
dissolve it in 50 mL of glacial acetic acid in a 250-mL flask. Transfer 50.0 mL of this solution to a glass-stoppered,
Add 2 drops of crystal violet TS, and titrate with the per- 500-mL flask, dilute with 50 mL of water, add 10 mL of po-
chloric acid solution until the violet color changes to blue- tassium iodide TS and 10 mL of dilute hydrochloric acid,
green. Deduct the volume of the perchloric acid consumed and allow to stand for 1 minute. Then add 15 mL of zinc
by 50 mL of the glacial acetic acid. Each 20.423 mg of po- sulfate solution (1 in 10), and titrate the liberated iodine
tassium biphthalate is equivalent to 1 mL of 0.1 N perchloric with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS
acid. as the endpoint is approached.
Protect from light, and restandardize before use.
Potassium Arsenite, Tenth-Normal (0.1 N)
KAsO2, 146.02
Potassium Hydroxide, Alcoholic, Half-Normal (0.5 N)
7.301 g in 1000 mL
28.06 g in 1000 mL
Dissolve 4.9455 g of arsenic trioxide primary standard,
Dissolve about 34 g of potassium hydroxide in 20 mL of
previously dried at 105 for 1 hour, in 75 mL of 1 N potas-
water, and add aldehyde-free alcohol to make 1000 mL. Al-
sium hydroxide. Add 40 g of potassium bicarbonate, dis-
low the solution to stand in a tightly stoppered bottle for
solved in about 200 mL of water, and dilute with water to
24 hours. Then quickly decant the clear supernatant into a
1000.0 mL.
suitable, tight container, and standardize the solution as
follows.
Potassium Bromate, Tenth-Normal (0.1 N)
Accurately measure about 25 mL of 0.5 N hydrochloric
KBrO3, 167.00
acid VS. Dilute with 50 mL of water, add 2 drops of phenol-
2.784 g in 1000 mL
phthalein TS, and titrate with the alcoholic potassium hy-
Dissolve 2.784 g of potassium bromate in water to make
droxide solution until a permanent, pale pink color is pro-
duced.
Transfer an accurately measured volume of about 40 mL
[NOTEStore in tightly stoppered bottles, protected from
of the solution to a glass-stoppered flask, add 3 g of potas-
light.]
sium iodide, and follow with 3 mL of hydrochloric acid. Al-
low to stand for 5 minutes, then titrate the liberated iodine
with 0.1 N sodium thiosulfate VS, adding 3 mL of starch TS
as the endpoint is approached. Correct for a blank run on
the same quantities of the same reagents, and calculate the
normality.
Potassium Hydroxide, Alcoholic, Tenth-Molar (0.1 M)
KOH, 56.11
Dilute 20 mL of 0.5 M alcoholic potassium hydroxide to
100.0 mL with aldehyde-free alcohol.
Potassium Hydroxide, Methanolic, Tenth-Normal
Potassium BromideBromate, Tenth-Normal (0.1 N)
(0.1 N)
Dissolve 2.78 g of potassium bromate (KBrO3) and 12.0 g
5.612 g in 1000 mL
of potassium bromide (KBr) in water, and dilute with water
Dissolve about 6.8 g of potassium hydroxide in 4 mL of
to 1000 mL. Standardize by the procedure set forth for Po-
water, and add methanol to make 1000 mL. Allow the solu-
tassium Bromate, Tenth-Normal (0.1 N).
tion to stand in a tightly stoppered bottle for 24 hours.
Then quickly decant the clear supernatant into a suitable,
tight container, and standardize the solution as follows.
Accurately measure about 25 mL of 0.1 N hydrochloric
acid VS. Dilute with 50 mL of water, add 2 drops of phenol-
phthalein TS, and titrate with the methanolic potassium hy-
droxide solution until a permanent, pale pink color is pro-
Potassium Dichromate, Tenth-Normal (0.1 N)
duced.
K2Cr2O7, 294.18
[NOTEStore in tightly stoppered bottles, protected from
4.903 g in 1000 mL
light.]
Dissolve about 5 g of potassium dichromate in 1000 mL
of water. Standardize the solution as follows.
Transfer 25.0 mL of this solution to a glass-stoppered,
500-mL flask, add 2 g of potassium iodide (free from io-
date), dilute with 200 mL of water, add 5 mL of hydrochlo-
ric acid, allow to stand for 10 minutes in a dark place, and
titrate the liberated iodine with 0.1 N sodium thiosulfate VS,
Potassium Hydroxide, Normal (1 N)
adding 3 mL of starch TS as the endpoint is approached.
KOH, 56.11
Carry out a blank determination.
56.11 g in 1000 mL
Dissolve 68 g of potassium hydroxide in about 950 mL of
water. Add a freshly prepared saturated solution of barium
hydroxide until no more precipitate forms. Shake the mix-
ture thoroughly, and allow it to stand overnight in a stop-
pered bottle. Decant the clear liquid, or filter the solution in
a tight, polyolefin bottle, and standardize by the procedure Silver Nitrate, Tenth-Normal (0.1 N)
set forth for Sodium Hydroxide, Normal (1 N). AgNO3, 169.87
16.99 g in 1000 mL
Dissolve about 17.5 g of silver nitrate in 1000 mL of
water, and standardize the solution as follows.
Transfer about 100 mg, accurately weighed, of reagent-
grade sodium chloride, previously dried at 110 for 2 hours,
to a 150-mL beaker, dissolve in 5 mL of water, and add
5 mL of acetic acid, 50 mL of methanol, and about 0.5 mL
Potassium Iodate, Twentieth-Molar (0.05 M)
of eosin Y TS. Stir, preferably with a magnetic stirrer, and
KIO3, 214.00
titrate with the silver nitrate solution.
10.70 g in 1000 mL
Dissolve 10.700 g of potassium iodate, previously dried at
110 to constant weight, in water to make 1000.0 mL. Stan-
dardize the solution as follows: to 15.0 mL of solution in a
250-mL iodine flask, add 3 g of potassium iodide and 3 mL
of hydrochloric acid previously diluted with 10 mL of water.
Stopper immediately, and allow to stand in the dark for
5 minutes. Then add 50 mL of cold water, and titrate the Sodium Arsenite, Twentieth-Molar (0.05 M)
liberated iodine with freshly standardized 0.1 N sodium thi- NaAsO2 129.91
osulfate. Add 3 mL of starch indicator solution near the end 6.496 g in 1000 mL
of the titration, and continue to the absence of the blue- Transfer 4.9455 g of arsenic trioxide, which has been pul-
starch-iodine complex. verized and dried at 100 to constant weight, to a 1000-mL
volumetric flask, dissolve it in 40 mL of 1 N sodium hydrox-
ide, and add 1 N sulfuric acid or 1 N hydrochloric acid until
the solution is neutral or only slightly acid to litmus. Add
30 g of sodium bicarbonate, dilute with water to volume,
and mix.
Potassium Permanganate, Tenth-Normal (0.1 N) Sodium Hydroxide, Alcoholic, Tenth-Normal (0.1 N)
KMnO4, 158.03 NaOH, 40.00
3.161 g in 1000 mL To 250 mL of alcohol add 2 mL of a 50% (w/v) solution
Dissolve about 3.3 g of potassium permanganate in of sodium hydroxide.
1000 mL of water in a flask, and boil the solution for about Dissolve about 200 mg of benzoic acid, accurately
15 minutes. Insert the stopper in the flask, allow it to stand weighed, in 10 mL of alcohol and 2 mL of water. Add
for at least 2 days, and filter through a fine-porosity, 2 drops of phenolphthalein TS, and titrate with the alcoholic
sintered-glass crucible. If necessary, the bottom of the sodium hydroxide solution until a permanent pale pink color
sintered-glass crucible may be lined with a pledget of glass is produced.
wool. Standardize the solution as follows.
Accurately weigh about 200 mg of sodium oxalate, dried
according to the instructions on its label, and dissolve it in
250 mL of water. Add 7 mL of sulfuric acid, heat to about
70, and then slowly add the permanganate solution from a
buret, with constant stirring, until a pale pink color, which
persists for 15 seconds, is produced. The temperature at the
Sodium Hydroxide, Normal (1 N)
conclusion of the titration should be not less than 60. Cal-
NaOH, 40.00
culate the normality. Each 6.700 mg of sodium oxalate is
40.00 g in 1000 mL
equivalent to 1 mL of 0.1 N potassium permanganate.
Dissolve 162 g of sodium hydroxide in 150 mL of carbon
Since potassium permanganate is reduced on contact
dioxide-free water, cool the solution to room temperature,
with organic substances such as rubber, the solution must
and filter through hardened filter paper. Transfer 54.5 mL of
be handled in apparatus entirely of glass or other suitably
the clear filtrate to a tight, polyolefin container, and dilute
inert material. It should be frequently restandardized. Store
with carbon dioxide-free water to 1000 mL.
in glass-stoppered, amber-colored bottles.
Accurately weigh about 5 g of potassium biphthalate, pre-
viously crushed lightly and dried at 120 for 2 hours, and
dissolve in 75 mL of carbon dioxide-free water. Add 2 drops
of phenolphthalein TS, and titrate with the sodium hydrox-
ide solution to the production of a permanent pink color.
Each 204.22 mg of potassium biphthalate is equivalent to
1 mL of 1 N sodium hydroxide.
Potassium Thiocyanate, Tenth-Normal (0.1 N)
KSCN, 97.18
9.72 g in 1000 mL
Weigh exactly 9.72 g of potassium thiocyanate, previously
[NOTES(1) Solutions of alkali hydroxides absorb carbon
dried for 2 hours at 110, transfer to a 1-L volumetric flask,
dioxide when exposed to air. They should be preserved in
dilute with water to volume, and mix well. Standardize as
bottles having well-fitted, suitable stoppers, provided with a
follows: transfer 40.0 mL of freshly standardized 0.1 N silver
tube filled with a mixture of sodium hydroxide and lime
nitrate VS to a 250-mL Erlenmeyer flask; and add 100 mL of
(soda-lime tubes) so that air entering the container must
water, 1 mL of nitric acid, and 2 mL of ferric ammonium
pass through this tube, which will absorb the carbon diox-
sulfate TS. Titrate with the potassium thiocyanate solution,
ide. (2) Prepare solutions of lower concentration (e.g.,
with agitation, to a permanent light pinkish-brown color of
0.1 N, 0.01 N) by quantitatively diluting accurately meas-
the supernatant.
ured volumes of the 1 N solution with sufficient carbon di-
oxide-free water to yield the desired concentration.]
Restandardize the solution frequently.
Sodium Methoxide, Half-Normal (0.5 N) in Methanol 1 mL of the endpoint, add the titrant in 0.1-mL portions,
CH3ONa, 54.02 and allow 1 minute between additions. Each 17.22 mg of
27.01 g in 1000 mL sulfanilamide is equivalent to 1 mL of 0.1000 M sodium
Weigh 11.5 g of freshly cut sodium metal, and cut into nitrite.
small cubes. Place about 0.5 mL of anhydrous methanol in a
round-bottom, 250-mL flask equipped with a ground-glass
joint, add 1 cube of the sodium metal, and, when the reac-
tion has ceased, add the remaining sodium metal to the
flask. Connect a water-jacketed condenser to the flask, and
slowly add 250 mL of anhydrous methanol, in small por-
tions, through the top of the condenser. Regulate the addi-
Sodium Tetraphenylboron, Fiftieth-Molar (0.02 M)
tion of the methanol so that the vapors are condensed and
NaB(C6H5)4, 342.22
do not escape through the top of the condenser. After addi-
6.845 g in 1000 mL
tion of the methanol is complete, connect a drying tube to
Dissolve an amount of sodium tetraphenylboron, equiva-
the top of the condenser, and allow the solution to cool.
lent to 6.845 g of NaB(C6H5)4, in water to make 1000 mL,
Transfer the solution to a 1-L volumetric flask, dilute with
and standardize the solution as follows.
anhydrous methanol to volume, and mix. Standardize the
Pipet two 75-mL portions of the solution into separate
beakers, and to each add 1 mL of acetic acid and 25 mL of
Accurately measure about 20 mL of freshly standardized
water. To each beaker add, slowly and with constant stir-
1 N hydrochloric acid VS into a 250-mL conical flask, add
ring, 25 mL of potassium biphthalate solution (1 in 20), and
0.25 mL of phenolphthalein TS, and titrate with the sodium
allow to stand for 2 hours. Filter one of the mixtures
methoxide solution to the first appearance of a permanent
through a filtering crucible, and wash the precipitate with
pink color.
cold water. Transfer the precipitate to a container, add
50 mL of water, shake intermittently for 30 minutes, filter,
and use the filtrate as the saturated potassium tetraphenyl-
borate solution in the following standardization procedure.
Filter the second mixture through a tared filtering crucible,
and wash the precipitate with three 5-mL portions of satu-
rated potassium tetraphenylborate solution. Dry the precipi-
Sodium Methoxide, Tenth-Normal (0.1 N) in Toluene tate at 105 for 1 hour. Each g of potassium tetraphenylbo-
CH3ONa, 54.02 rate (KTPB) is equivalent to 955.1 mg of sodium
5.402 g in 1000 mL tetraphenylboron.
Cool in ice-water 150 mL of methanol contained in a
1000-mL volumetric flask, and add, in small portions, about
2.5 g of freshly cut sodium metal. When the metal has dis-
solved, add toluene to make 1000 mL, and mix. Store pref-
erably in the reservoir of an automatic delivery buret suita-
bly protected from carbon dioxide and moisture. [NOTEPrepare this solution just before use.]
Accurately weigh about 400 mg of primary standard ben-
Sodium Thiosulfate, Tenth-Normal (0.1 N)
zoic acid, and dissolve in 80 mL of dimethylformamide in a
Na2S2O3 5H2O, 248.19
flask. Add 3 drops of a 1 in 100 solution of thymol blue in
24.82 g in 1000 mL
dimethylformamide, and titrate with the sodium methoxide
Dissolve about 26 g of sodium thiosulfate and 200 mg of
to a blue endpoint. Correct for the volume of the sodium
sodium carbonate in 1000 mL of recently boiled and cooled
methoxide solution consumed by 80 mL of the dimethyl-
water. Standardize the solution as follows.
formamide. Each 12.21 mg of benzoic acid is equivalent to
Accurately weigh about 210 mg of primary standard po-
1 mL of 0.1 N sodium methoxide.
tassium dichromate, previously pulverized and dried accord-
ing to the instructions on its label, if necessary, and dissolve
in 100 mL of water in a glass-stoppered, 500-mL flask. Swirl
to dissolve the solid, remove the stopper, and quickly add
3 g of potassium iodide, 2 g of sodium bicarbonate, and
5 mL of hydrochloric acid. Insert the stopper gently in the
[NOTES(1) To eliminate any turbidity that may form fol-
flask, swirl to mix, and allow to stand in the dark for exactly
lowing dilution with toluene, add methanol (25 to 30 mL
10 minutes. Rinse the stopper and the inner walls of the
usually suffices) until the solution is clear. (2) Restandardize
flask with water, and titrate the liberated iodine with the
the solution frequently.]
sodium thiosulfate solution until the solution is yellowish
green in color. Add 3 mL of starch TS, and continue the
Sodium Nitrite, Tenth-Molar (0.1 M)
titration until the blue color is discharged. Perform a blank
NaNO2, 69.00
determination.
6.900 g in 1000 mL
Restandardize the solution as frequently as supported by
Dissolve 7.5 g of sodium nitrite in water to make
laboratory stability data. In the absence of such data,
restandardize the solution weekly.
Accurately weigh about 500 mg of USP Sulfanilamide RS,
previously dried at 105 for 3 hours, and transfer to a suita-
ble beaker. Add 20 mL of hydrochloric acid and 50 mL of
water, stir until dissolved, and cool to 15. Maintaining the
temperature at about 15, titrate slowly with the sodium
nitrite solution, placing the buret tip below the surface of
the solution to preclude air oxidation of the sodium nitrite,
Sulfuric Acid, Half-Normal (0.5 N) in Alcohol
and stir the solution gently with a magnetic stirrer, but
H2SO4, 98.08
avoid pulling a vortex of air beneath the surface. Use the
24.52 g in 1000 mL
indicator specified in the individual monograph, or, if a
Add slowly, with stirring, 13.9 mL of sulfuric acid to a
potentiometric procedure is specified, determine the
sufficient quantity of dehydrated alcohol to make 1000 mL.
endpoint electrometrically, using platinumcalomel or pla-
tinumplatinum electrodes. When the titration is within
Cool, and standardize against tromethamine as described Tetramethylammonium Bromide, Tenth-Molar (0.1 M)
under Hydrochloric Acid, Half-Normal (0.5 N) in Methanol. (CH3)4NBr, 154.05
15.41 g in 1000 mL
Dissolve 15.41 g of tetramethylammonium bromide in
water to make 1000 mL, and standardize the solution as
follows.
of the solution to a beaker, add 10 mL of diluted nitric acid
and 50.0 mL of 0.1 N silver nitrate VS, and mix. Add 2 mL
Sulfuric Acid, Normal (1 N)
of ferric ammonium sulfate TS, and titrate the excess silver
H2SO4, 98.08
nitrate with 0.1 N ammonium thiocyanate VS.
49.04 g in 1000 mL
Add slowly, with stirring, 27 mL of sulfuric acid to a suffi-
cient quantity of water to make 1000 mL. Cool and stan-
dardize against tromethamine as described under Hydrochlo-
ric Acid, Normal (1 N).
Tetrabutylammonium Hydroxide, Tenth-Normal (0.1 N)
Tetramethylammonium Chloride, Tenth-Molar (0.1 M)
(C4H9)4NOH, 259.47
(CH3)4NCl, 109.60
25.95 g in 1000 mL
10.96 g in 1000 mL
Dissolve 40 g of tetra-n-butylammonium iodide in 90 mL
Dissolve 10.96 g of tetramethylammonium chloride in
of anhydrous methanol in a glass-stoppered flask. Place in
water to make 1000 mL, and standardize the solution as
an ice bath, add 20 g of powdered silver oxide, insert the
follows.
stopper in the flask, and agitate vigorously for 60 minutes.
Centrifuge a few mL, and test the supernatant for iodide
of the solution to a flask, add 10 mL of diluted nitric acid
(see Iodide 191). If the test is positive, add an additional
and 50.0 mL of 0.1 N silver nitrate VS, and mix. Add 5 mL
2 g of silver oxide, and continue to allow to stand for
of nitrobenzene and 2 mL of ferric ammonium sulfate TS,
30 minutes with intermittent agitation. When all of the io-
shake, and titrate the excess silver nitrate with 0.1 N ammo-
dide has reacted, filter through a fine-porosity, sintered-glass
nium thiocyanate VS.
funnel. Rinse the flask and the funnel with three 50-mL por-
tions of anhydrous toluene, adding the rinsings to the fil-
trate. Dilute with a mixture of three volumes of anhydrous
toluene and 1 volume of anhydrous methanol to 1000 mL,
and flush the solution for 10 minutes with dry, carbon diox-
ide-free nitrogen. [NOTEIf necessary to obtain a clear solu-
tion, further small quantities of anhydrous methanol may be
added.] Store in a reservoir protected from carbon dioxide Titanium Trichloride, Tenth-Normal (0.1 N)
and moisture, and discard after 60 days. Alternatively, the TiCl3, 154.23
solution may be prepared by diluting a suitable volume of 15.42 g in 1000 mL
commercially available tetrabutylammonium hydroxide solu- Add 75 mL of titanium trichloride solution (1 in 5) to
tion in methanol with a mixture of 4 volumes of anhydrous 75 mL of hydrochloric acid, dilute to 1000 mL, and mix.
toluene and 1 volume of anhydrous methanol. [NOTEIf Standardize the solution as follows, using the special titra-
necessary to obtain a clear solution, further small quantities tion apparatus described.
of methanol may be added.] ApparatusStore the titanium trichloride solution in the
Standardize the solution on the day of use as follows. reservoir of a closed-system titration apparatus in an atmos-
Dissolve about 400 mg of primary standard benzoic acid, phere of hydrogen.
accurately weighed, in 80 mL of dimethylformamide, add Use a wide-mouth, 500-mL conical flask as the titration
3 drops of a 1 in 100 solution of thymol blue in dimethyl- vessel, and connect it by means of a tight-fitting rubber
formamide, and titrate to a blue endpoint with the tetrabu- stopper to the titration buret, an inlet tube for carbon diox-
tylammonium hydroxide solution, delivering the titrant from ide, and an exit tube. Arrange for mechanical stirring. All
a buret equipped with a carbon dioxide absorption trap. joints must be air-tight. Arrange to have both the hydrogen
Perform a blank determination, and make any necessary and the carbon dioxide pass through wash bottles contain-
correction. Each mL of 0.1 N tetrabutylammonium hydrox- ing titanium trichloride solution (approximately 1 in 50) to
ide is equivalent to 12.21 mg of benzoic acid. remove any oxygen.
If the solution to be titrated is to be heated before or
during titration, connect the titration flask with an upright
reflux condenser through the rubber stopper.
StandardizationPlace an accurately measured volume of
about 40 mL of 0.1 N ferric ammonium sulfate VS in the
titration flask, and pass in a rapid stream of carbon dioxide
until all the air has been removed. Add the titanium trichlo-
Tetrabutylammonium Hydroxide in Methanol/
ride solution from the buret until near the calculated
Isopropyl Alcohol, 0.1 N
endpoint (about 35 mL), then add through the outlet tube
Prepare as described for Tetrabutylammonium Hydroxide,
5 mL of ammonium thiocyanate TS, and continue the titra-
Tenth-Normal (0.1 N) using isopropyl alcohol instead of tolu-
tion until the solution is colorless.
ene, and standardize as described. Alternatively, the solution
may be prepared by diluting a suitable volume of commer-
cially available tetrabutylammonium hydroxide solution in
methanol with 4 volumes of anhydrous isopropyl alcohol.
Zinc Sulfate, Twentieth-Molar (0.05 M)
ZnSO4 7H2O, 287.56
14.4 g in 1000 mL
USP 36 Reagents / Chromatographic Columns 1227
Dissolve 14.4 g of zinc sulfate in water to make 1 L. Stan- 50 mL of alcohol, and 2 mL of dithizone TS. Titrate with the
dardize the solution as follows. zinc sulfate solution to a clear, rose-pink color.
Accurately measure about 10 mL of 0.05 M edetate diso-
dium VS into a 125-mL conical flask, and add, in the order
given, 10 mL of acetic acidammonium acetate buffer TS,
Chromatographic Columns
The following list of packings (L), phases (G), and sup- L19Strong cation-exchange resin consisting of
ports (S) is intended to be a convenient reference for the sulfonated cross-linked styrene-divinylbenzene copolymer in
chromatographer. [NOTEParticle sizes given in this listing the calcium form, about 9 m in diameter.
are those generally provided. Where other, usually finer, L20Dihydroxypropane groups chemically bonded to po-
sizes are required, the individual monograph specifies the rous silica or hybrid particles, 1.5 to 10 m in diameter.
desired particle size. Within any category of packings or L21A rigid, spherical styrene-divinylbenzene copolymer
phases listed below, there may be a wide range of columns 3 to 10 m in diameter.
available. Where it is necessary to define more specifically L22A cation-exchange resin made of porous polystyrene
the chromatographic conditions, the individual monograph gel with sulfonic acid groups, about 10 m in size.
so indicates.] L23An anion-exchange resin made of porous
polymethacrylate or polyacrylate gel with quaternary ammo-
nium groups, 712 m in size.
Packings
L24A semi-rigid hydrophilic gel consisting of vinyl
polymers with numerous hydroxyl groups on the matrix sur-
L1Octadecyl silane chemically bonded to porous or
face, 32 to 63 m in diameter. [NOTEAvailable as YMC-
nonporous silica or ceramic microparticles, 1.5 to 10 m in
Pack PVA-SIL manufactured by YMC Co., Ltd. and distrib-
diameter, or a monolithic silica rod.
uted by Waters Corp. (www.waters.com).]
L2Octadecyl silane chemically bonded to silica gel of a
L25Packing having the capacity to separate compounds
controlled surface porosity that has been bonded to a solid
with a molecular weight range from 1005000 (as deter-
spherical core, 30 to 50 m in diameter.
mined by polyethylene oxide), applied to neutral, anionic,
L3Porous silica particles, 1.5 to 10 m in diameter, or a
and cationic water-soluble polymers. A polymethacrylate
monolithic silica rod.
resin base, cross-linked with polyhydroxylated ether (surface
L4Silica gel of controlled surface porosity bonded to a
contained some residual carboxyl functional groups) was
solid spherical core, 30 to 50 m in diameter.
found suitable.
L5Alumina of controlled surface porosity bonded to a
L26Butyl silane chemically bonded to totally porous sil-
solid spherical core, 30 to 50 m in diameter.
ica particles, 1.5 to 10 m in diameter.
L6Strong cation-exchange packingsulfonated fluoro-
L27Porous silica particles, 30 to 50 m in diameter.
carbon polymer coated on a solid spherical core, 30 to
L28A multifunctional support, which consists of a high
50 m in diameter.
purity, 100
A, spherical silica substrate that has been

L7Octylsilane chemically bonded to totally porous or
bonded with anionic exchanger, amine functionality in addi-
superficially porous silica particles, 1.510 m in diameter,
tion to a conventional reversed phase C8 functionality.
or a monolithic silica rod.
L29Gamma alumina, reverse-phase, low carbon per-
L8An essentially monomolecular layer of aminopropylsi-
centage by weight, alumina-based polybutadiene spherical
lane chemically bonded to totally porous silica gel support,
particles, 5 m in diameter with a pore volume of 80
A.
1.5 to 10 m in diameter.
L30Ethyl silane chemically bonded to totally porous sil-
L9Irregular or spherical, totally porous silica gel having
ica particles, 3 to 10 m in diameter.
a chemically bonded, strongly acidic cation-exchange coat-
L31A hydroxide-selective, strong anion-exchange resin-
ing, 3 to 10 m in diameter.
quaternary amine bonded on latex particles attached to a
L10Nitrile groups chemically bonded to porous silica
core of 8.5-m macroporous particles having a pore size of
particles, 1.5 to 10 m in diameter.
2000
A and consisting of ethylvinylbenzene cross-linked

L11Phenyl groups chemically bonded to porous silica
with 55% divinylbenzene.
particles, 1.5 to 10 m in diameter.
L32A chiral ligand-exchange packingL-proline copper
L12A strong anion-exchange packing made by chemi-
complex covalently bonded to irregularly shaped silica parti-
cally bonding a quaternary amine to a solid silica spherical
cles, 5 to 10 m in diameter.
core, 30 to 50 m in diameter.
L33Packing having the capacity to separate dextrans by
L13Trimethylsilane chemically bonded to porous silica
molecular size over a range of 4,000 to 500,000 Da. It is
particles, 3 to 10 m in diameter.
spherical, silica-based, and processed to provide pH stability.
L14Silica gel having a chemically bonded, strongly basic
[NOTEAvailable as TSK-GEL G4000SWxl from Tosoh Bio-
quaternary ammonium anion-exchange coating, 5 to 10 m
science (www.tosohbioscience.com).]
in diameter.
L34Strong cation-exchange resin consisting of
L15Hexylsilane chemically bonded to totally porous sil-
sulfonated cross-linked styrenedivinylbenzene copolymer in
ica particles, 3 to 10 m in diameter.
the lead form, 7 to 9 m in diameter.
L16Dimethylsilane chemically bonded to porous silica
L35A zirconium-stabilized spherical silica packing with a
particles, 5 to 10 m in diameter.
hydrophilic (diol-type) molecular monolayer bonded phase
L17Strong cation-exchange resin consisting of
having a pore size of 150
A.
sulfonated cross-linked styrenedivinylbenzene copolymer in
L36A 3,5-dinitrobenzoyl derivative of L-phenylglycine
the hydrogen form, 6 to 12 m in diameter.
covalently bonded to 5-m aminopropyl silica.
L18Amino and cyano groups chemically bonded to po-
L37Packing having the capacity to separate proteins by
rous silica particles, 3 to 10 m in diameter.
molecular size over a range of 2,000 to 40,000 Da. It is a
polymethacrylate gel.
1228 Chromatographic Columns / Reagents USP 36
L38A methacrylate-based size-exclusion packing for the sodium form, about 6 to 30 m in diameter. [NOTE
water-soluble samples. Available as Aminex HPX-87N from Bio-Rad Laboratories,
L39A hydrophilic polyhydroxymethacrylate gel of totally (2000/01 catalog, #125-0143) www.bio-rad.com.]
porous spherical resin. L59Packing for the size-exclusion separation of proteins
L40Cellulose tris-3,5-dimethylphenylcarbamate coated (separation by molecular weight) over the range of 5 to
porous silica particles, 5 to 20 m in diameter. 7000 kDa. The packing is a spherical 1.5- to 10-m silica or
L41Immobilized 1-acid glycoprotein on spherical silica hybrid packing with a hydrophilic coating.
particles, 5 m in diameter. L60Spherical, porous silica gel, 10 m or less in diame-
L42Octylsilane and octadecylsilane groups chemically ter, the surface of which has been covalently modified with
bonded to porous silica particles, 5 m in diameter. alkyl amide groups and endcapped. [NOTEAvailable as
L43Pentafluorophenyl groups chemically bonded to sil- Supelcosil LC-ABZ from Supelco (www.sigmaaldrich.com/
ica particles by a propyl spacer, 1.5 to 10 m in diameter. supelco).]
L44A multifunctional support, which consists of a high L61A hydroxide selective strong anion-exchange resin
purity, 60
A, spherical silica substrate that has been bonded consisting of a highly cross-linked core of 13-m micro-
with a cationic exchanger, sulfonic acid functionality in ad- porous particles having a pore size less than 10
A units and
dition to a conventional reversed phase C8 functionality. consisting of ethylvinylbenzene cross-linked with 55%
L45Beta cyclodextrin bonded to porous silica particles, divinylbenzene with a latex coating composed of 85-nm di-
5 to 10 m in diameter. ameter microbeads bonded with alkanol quaternary ammo-
L46Polystyrene/divinylbenzene substrate agglomerated nium ions (6%). [NOTEAvailable as Ion Pac AS-11 and AG-
with quaternary amine functionalized latex beads, about 9 11 from Dionex (www.dionex.com).]
to 11 m in diameter. L62C30 silane bonded phase on a fully porous spherical
L47High-capacity anion-exchange microporous sub- silica, 3 to 15 m in diameter.
strate, fully functionalized with trimethlyamine groups, 8 m L63Glycopeptide teicoplanin linked through multiple
in diameter. [NOTEAvailable as CarboPac MA1 and distrib- covalent bonds to a 100-
A units spherical silica.

uted by Dionex Corp. (www.dionex.com).] [NOTEAvailable as Astec Chirobiotic T from Supelco (www.
L48Sulfonated, cross-linked polystyrene with an outer sigmaaldrich.com).]
layer of submicron, porous, anion-exchange microbeads, 10 L64Strongly basic anion-exchange resin consisting of
to 15 m in diameter. 8% cross-linked styrene-divinylbenzene copolymer with a
L49A reversed-phase packing made by coating a thin quaternary ammonium group in the chloride form, 45 to
layer of polybutadiene onto spherical porous zirconia parti- 180 m in diameter. [NOTEA suitable grade is available as
cles, 3 to 10 m in diameter. [NOTEAvailable as Zirchrom AG 1-X8 resin chloride form from www.discover.bio-rad.
PBD from www.zirchrom.com.] com.]
L50Multifunction resin with reversed-phase retention L65Strongly acidic cation-exchange resin consisting of
and strong anion-exchange functionalities. The resin consists 8% sulfonated cross-linked styrene-divinylbenzene co-
of ethylvinylbenzene, 55% cross-linked with divinylbenzene polymer with a sulfonic acid group in the hydrogen form,
copolymer, 3 to 15 m in diameter, and a surface area not 45 to 250 m in diameter. [NOTEA suitable grade is availa-
less than 350 m
2
per g. Substrate is coated with quaternary ble as AG 50W-X2 resin hydrogen form from
ammonium functionalized latex particles consisting of sty- www.discover.bio-rad.com.]
rene cross-linked with divinylbenzene. [NOTEAvailable as L66A crown ether coated on a 5-m particle size silica
OmniPac PAX-500 and distributed by Dionex Corp. (www. gel substrate. The active site is (S)-18-crown-6-ether.
dionex.com).] [NOTEAvailable as Crownpak CR(+) from Daicel
L51Amylose tris-3,5-dimethylphenylcarbamate-coated, (www.daicel.com).]
porous, spherical, silica particles, 5 to 10 m in diameter. L67Porous vinyl alcohol copolymer with a C18 alkyl
[NOTEAvailable as Chiralpak AD from Chiral Technologies, group attached to the hydroxyl group of the polymer, 2 to
Inc., (www.chiraltech.com).] 10 m in diameter. [NOTEAvailable as apHera C18 from
L52A strong cation-exchange resin made of porous sil- Supelco (www.sigmaaldrich.com).]
ica with sulfopropyl groups, 5 to 10 m in diameter. L68Spherical, porous silica, 10 m or less in diameter,
[NOTEAvailable as TSK-GEL IC-Cation-SW from Tosoh Bio- the surface of which has been covalently modified with alkyl
science (www.tosohbioscience.com).] amide groups and not endcapped. [NOTEAvailable as
L53Weak cation-exchange resin consisting of SUPELCOSIL SUPLEX pKb-100 from Supelco
ethylvinylbenzene, 55% cross-linked with divinylbenzene co- (www.sigmaaldrich.com).]
polymer, 3 to 15 m diameter. Substrate is surface grafted L69Ethylvinylbenzene/divinylbenzene substrate agglom-
with carboxylic acid and/or phosphoric acid functionalized erated with quaternary amine functionalized 130-nm latex
monomers. Capacity not less than 500 Eq/column. [NOTE beads, about 6.5 m in diameter. [NOTEAvailable as
Available as IonPac CS14 distributed by Dionex Corp. CarboPac PA20 from www.dionex.com.]
(www.dionex.com).] L70Cellulose tris(phenyl carbamate) coated on 5-m sil-
L54A size exclusion medium made of covalent bonding ica. [NOTEAvailable as Chiralcel OC-H from www.
of dextran to highly cross-linked porous agarose beads, chiraltech.com.]
about 13 m in diameter. L71A rigid, spherical polymetacrylate, 4 to 6 m in di-
[NOTEAvailable as Superdex Peptide HR 10/30 from ameter. [NOTEAvailable as RSpak DE-613 from www.
www.gelifesciences.com.] shodex.com.]
L55A strong cation-exchange resin made of porous sil- L72(S)-phenylglycine and 3,5-dinitroanaline urea link-
ica coated with polybutadienemaleic acid copolymer, age covalently bonded to silica.
about 5 m in diameter. [NOTEAvailable as IC-Pak C M/D [NOTEAvailable as Sumichiral OA-3300, distributed by
from Waters Corp. (www.waters.com).] www.phenomenex.com.]
L56Propyl silane chemically bonded to totally porous L73A rigid spherical polydivinylbenzene particle, 5 to
silica particles, 3 to 10 m in diameter. [NOTEAvailable as 10 m in diameter. [NOTEAvailable as Jordi-Gel DBV from
Zorbax SB-C3 from Agilent Technologies (www.agilent.com/ www.jordiflp.com.]
chem).] L74a strong anion-exchange resin consisting of a highly
L57A chiral-recognition protein, ovomucoid, chemically cross-linked core of 7-m macroporous particles having a
bonded to silica particles, about 5 m in diameter, with a 100-
A average pore size and consisting of ethylvinylbenzene

pore size of 120
A. [NOTEAvailable as Ultron ES-OVM cross-linked with 55% divinylbenzene and an anion-ex-

from Agilent Technologies (www.agilent.com/chem).] change layer grafted to the surface, which is functionalized
L58Strong cation-exchange resin consisting of with alkyl quaternary ammonium ions. [NOTEAvailable as
sulfonated cross-linked styrene-divinylbenzene copolymer in IonPac AS14A from Dionex (www.dionex.com).]
USP 36 Reagents / Chromatographic Columns 1229
L75A chiral-recognition protein, bovine serum albumine G39Polyethylene glycol (av. mol. wt. about 1500).
(BSA), chemically bonded to silica particles, about 7 m in G40Ethylene glycol adipate.
diameter, with a pore size of 300
A. G41Phenylmethyldimethylsilicone (10% phenyl-substi-

tuted).
G4235% phenyl-65% dimethylpolysiloxane (percent-
Phases
ages refer to molar substitution).
G436% cyanopropylphenyl-94% dimethylpolysiloxane
G1Dimethylpolysiloxane oil.
(percentages refer to molar substitution).
G2Dimethylpolysiloxane gum.
G442% low molecular weight petrolatum hydrocarbon
G350% Phenyl-50% methylpolysiloxane.
grease and 1% solution of potassium hydroxide.
G4Diethylene glycol succinate polyester.
G45Divinylbenzene-ethylene glycol-dimethylacrylate.
G53-Cyanopropylpolysiloxane.
G4614% Cyanopropylphenyl-86% methylpolysiloxane.
G6Trifluoropropylmethylpolysiloxane.
G47Polyethylene glycol (av. mol. wt. of about 8000).
G750% 3-Cyanopropyl-50% phenylmethylsilicone.
G48Highly polar, partially cross-linked cyanopolysilox-
G880% Bis(3-cyanopropyl)-20% 3-cya-
ane.
nopropylphenylpoly-siloxane (percentages refer to molar
substitution).
G9Methylvinylpolysiloxane. Supports
G10Polyamide formed by reacting a C36 dicarboxylic
acid with 1,3-di-4-piperidylpropane and piperidine in the re- [NOTEUnless otherwise specified, mesh sizes of 80 to
spective mole ratios of 1.00:0.90:0.20. 100 or, alternatively, 100 to 120 are intended.]
G11Bis(2-ethylhexyl) sebacate polyester. S1ASiliceous earth for gas chromatography has been
G12Phenyldiethanolamine succinate polyester. flux-calcined by mixing diatomite with Na2CO3 flux and cal-
G13Sorbitol. cining above 900. The siliceous earth is acid-washed, then
G14Polyethylene glycol (av. mol. wt. of 950 to 1050). water-washed until neutral, but not base-washed. The sili-
G15Polyethylene glycol (av. mol. wt. of 3000 to 3700). ceous earth may be silanized by treating with an agent such
G16Polyethylene glycol compound (av. mol. wt. about as dimethyldichlorosilane [NOTEUnless otherwise specified
15,000). A high molecular weight compound of polyethyl- in the individual monograph, silanized support is intended.]
ene glycol with a diepoxide linker. [NOTEAvailable com- to mask surface silanol groups.
mercially as Polyethylene Glycol Compound 20M, or as S1ABThe siliceous earth as described above is both
Carbowax 20M, from suppliers of chromatographic acid- and base-washed. [NOTEUnless otherwise specified in
reagents.] the individual monograph, silanized support is intended.]
G1775% Phenyl-25% methylpolysiloxane. S1CA support prepared from crushed firebrick and cal-
G18Polyalkylene glycol. cined or burned with a clay binder above 900 with subse-
G1925% Phenyl-25% cyanopropyl-50% methylsilicone. quent acid-wash. It may be silanized.
G20Polyethylene glycol (av. mol. wt. of 380 to 420). S1NSThe siliceous earth is untreated.
G21Neopentyl glycol succinate. S2Styrene-divinylbenzene copolymer having a nominal
G22Bis(2-ethylhexyl) phthalate. surface area of less than 50 m
2
per g and an average pore
G23Polyethylene glycol adipate. diameter of 0.3 to 0.4 m.
G24Diisodecyl phthalate. S3Copolymer of ethylvinylbenzene and divinylbenzene
G25Polyethylene glycol compound TPA. A high molecu- having a nominal surface area of 500 to 600 m
2
per g and
lar weight compound of a polyethylene glycol and a diepo- an average pore diameter of 0.0075 m.
xide that is esterified with terephthalic acid. S4Styrene-divinylbenzene copolymer with aromatic O
[NOTEAvailable commercially as Carbowax 20M-TPA and N groups, having a nominal surface area of 400 to
from suppliers of chromatographic reagents.] 600 m
2
per g and an average pore diameter of 0.0076 m.
G2625% 2-Cyanoethyl-75% methylpolysiloxane. S540- to 60-mesh, high-molecular weight tetrafluoreth-
G275% Phenyl-95% methylpolysiloxane. ylene polymer.
G2825% Phenyl-75% methylpolysiloxane. S6Styrene-divinylbenzene copolymer having a nominal
G293,3-Thiodipropionitrile. surface area of 250 to 350 m
2
per g and an average pore
G30Tetraethylene glycol dimethyl ether. diameter of 0.0091 m.
G31Nonylphenoxypoly(ethyleneoxy)ethanol (av. S7Graphitized carbon having a nominal surface area of
ethyleneoxy chain length is 30); Nonoxynol 30. 12 m
2
per g.
G3220% Phenylmethyl-80% dimethylpolysiloxane. S8Copolymer of 4-vinyl-pyridine and styrene-
G3320% Carborane-80% methylsilicone. divinylbenzene.
G34Diethylene glycol succinate polyester stabilized with S9A porous polymer based on 2,6-diphenyl-p-phenyl-
phosphoric acid. ene oxide.
G35A high molecular weight compound of a polyethyl- S10A highly polar cross-linked copolymer of acryloni-
ene glycol and a diepoxide that is esterified with nitroter- trite and divinylbenzene.
ephthalic acid. S11Graphitized carbon having a nominal surface area of
G361% Vinyl-5% phenylmethylpolysiloxane. 100 m
2
per g modified with small amounts of petrolatum
G37Polyimide. and polyethylene glycol compound.
G38Phase G1 containing a small percentage of a tailing S12Graphitized carbon having a nominal surface area of
inhibitor. 100 m
2
per g.
SP2100/0.1% Carbowax 1500 from Supelco, Inc. (www.
sigmaaldrich.com/supelco).]

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USP 36 Reagents / Reagents 1133

Reagents, Indicators and

167.64 [123-46-6]Color- acetone; insoluble in ether. Usually contains solvent of crys-

A, spherical silica substrate that has been

A and consisting of ethylvinylbenzene cross-linked

A units spherical silica.

A average pore size and consisting of ethylvinylbenzene

A. [NOTEAvailable as Ultron ES-OVM cross-linked with 55% divinylbenzene and an anion-ex-

A. G41Phenylmethyldimethylsilicone (10% phenyl-substi-

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