Purospher STAR LP - Application Guide

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany
http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com
Introduction
Purospher HPLC columns are based upon a high-purity, metal free silica for
excellent separations with good symmetry. The base material for Purospher highpurity HPLC columns is made from tetra-alkoxysilane. Due to the absence of metals
in the silica matrix and in combination with an excellent coverage of the silica
surface, this stationary phase enables tailing free chromatography of acidic, basic
and chelating compounds. This is of particular advantage for method development.
The newest addition to the Purospher family is Purospher STAR LP RP-18
endcapped. The packing material for Purospher STAR LP RP-18 endcapped has
been optimized on pressure which gives at least 25% less back pressure in
comparison with existing Purospher STAR RP-18 endcapped. This will give more
opportunity to develop method for formulated drugs.
The application guide will illustrate different applications with excellent separation
on formulated product like Tablet, Capsules, Injections etc.

02
Specification for Purospher STAR LP RP-18 endcapped

Sorbent:
Particle Size:
Pore Size:
Surface Area:
Carbon Load:
pH range:
Performance:
High purity silica gel

5m
120
330 m2/gm
17 %
1.5 to 10.5
> 70,000 N/m
Purospher STAR LP RP-18e Vs Competition

Back Pressure Comparison with MeOH/H2O 50:50 (v/v)
179
180
165
164
157
Back pressure (Bar)
160
140
120
100
80
60
40
20
0
Instrument 1
Instrument 2
Purospher STAR LP
250x4.6 mm, 5 m
179
164
Competition Column
250x4.6 mm, 5 m
165
157

03
Application Index
Formulation Name
Compound class
Therapeutic use
Atorvastatin Tablet
Statins
Lowering blood
cholesterol
Chlorocresol, Beclomethasone,
Miconazole Cream
Disinfectant
Bisoprolol, Amlodipine Tablet
Analysis
Methods
IP
Inflammatory skin
disorder
In-house
Beta blocker &

Antihypertensive
Cardiovascular
disease
In-house
Ciprofloxacin Eye drops
Antibiotic
Eye infection
USP/IP
Cyanocobalamine Injection
Vitamin B12
In-house
Fenofibrate Capsule
Fibrate
USP
10
Folic acid tablet
Vitamin B9
IP
11
Hydrochlorothiazide, Ramipril
Tablet
Antihypertensive
High blood pressure
In-house
12
Ivermectin Injection
Antiparasitic
River blindness
USP
13
Levofloxacin Tablet
Antibiotic
Bacterial infection
IP
14
Mecobalamine Injection
Vitamin
JP
15
Metformin, Pioglitazole Tablet
Antidiabetic
High blood sugar
In-house
16
Nebivolol Tablet
Antihypertensive
Cardiovascular
disease
IP
17
Ofloxacin Tablet
Antibiotic
Bacterial infection
IP
18
Olanzapine Tablet
Antipsychotic
Bipolar disorder
IP
19
Olmesartan, Hydrochlorothiazide
Tablet
Angiotensin II
receptor &
Antihypertensive
Cardiovascular
disease
In-house
20
Olmesartan, Amlodipine Tablet
Angiotensin II
receptor &
Antihypertensive
Cardiovascular
disease
In-house
21
Olmesratan Tablet
Angiotensin II
receptor
Cardiovascular
disease
In-house
22
Reduce cholesterol &

triglyceride

04
Purospher STAR LP Atorvastatin Tablet

Chromatographic Conditions
Time
(min)
Flow
(ml/min)
%B
%C
Column:
Purospher STAR LP RP18e 250x4.6 mm, 5.0 m
0.0
1.8
100
Injection:
20 L
20.0
1.8
100
Detection:
Shimadzu 2010 CHT, UV 246 nm
35.0
1.5
25
75
Cell:
10 L
40.0
1.5
25
75
Gradient:
See table
55.0
1.5
100
Mobile Phase (v/v):
A: Acetonitrile : Tetrahydrofuran (92.5:7.5 v/v)
60.0
1.8
100
70.0
1.8
100
B: 5.75 g of ammonium dihydogen phosphate in 1L water.

Mix Buffer & Mobile phase A 58:42 (v/v).
C: Buffer:Mobile phase A & Methanol 20:20:60 (v/v)
Temperature:
25 C
Diluent:
Acetonitrile: Water 40:60 (v/v)
Sample:
Crush 20 tablet. Weigh equivalent to 50 mg. of Atorvastatin. Disperse in 10 ml Methanol.

Add 20 ml Diluent: & sonicate for 30 min. Dilute to 100 ml & filter through 0.45 m filter paper.
Pressure Drop:
220 Bar to 200 Bar (3190 psi to 2900 psi)
1200
Atorvastatin
Intensity (mV)
1000
800
600
400
200
0
0
10
20
30
40
50
Retention Time (min)

Chromatographic Data
No.
1
Compound
Atorvastatin
Time (min)
Tailing Factor
Theoretical Plate
19.6
1.0
11033

05
Purospher STAR LP Beclomethasone, Chlorocresol, Miconazole

Column:
Injection:
Detection:
Cell:
Flow Rate:
Mobile Phase (v/v):
Temperature:
Diluent:
Standard:
Sample:
Pressure Drop:
Purospher STAR LP RP-18e 250x4.6 mm, 5 m

20 L
Shimadzu Prominence, UV 240nm
10 L
1.0 mL/min
6.8 g of Sodium acetate trihydrate in 1 L water. Adjust pH to 7.4 with dilute H2SO4
Mix Acetonitrile and Buffer 70:30 (v/v)
30 C
Methanol
Solution I: 50 mg Beclomethasone in 50 ml methanol
Solution II: 40 mg Chlorocresol in 100 ml volumetric flask. Add 35 ml methanol to dissolve.
Add 10 ml Solution I & make up the volume with methanol.
Solution III:40 mg Miconazole in 100 ml volumetric flask. Add 35 ml methanol to dissolve.
Add 5 ml Solution II & make up the volume with methanol.
2 g of cream in 100 ml volumetric flask. Add 35 ml of methanol & sonicate for 15 min.
Dilute to volume with methanol & filter through 0.45 m filter paper.
46 Bar (667 psi)
80
Beclomethasone
120
Miconazole
160
Chlorocresol
Intensity (mV)
200
40
0
0
10
15
20
25
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Chlorocresol
4.5
1.2
12210
Beclomethasone
8.6
1.1
13911
Miconazole
18.7
1.2
16398

06
Purospher STAR LP Bisoprolol Fumarate and Amlodipine Tablet

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
3.45 g Ammonium di-hydrogen phosphate in 1L water. Adjust pH to 3.0

with ortho-phosphoric acid . Mix Buffer & Acetonitrile 60:40 (v/v)
Temperature:
30 C
Diluent:
Mobile phase
Standard:
20 mg Bisoprolol & 56 mg amlodipine diluted to 200 ml with diluent.
Sample:
Weigh equivalent to two tablet. Add mobile phase to 50 ml.

Sonicate for 30 min & filter through 0.45 m filter paper.
Pressure Drop:
116 Bar (1682 psi)
600
Amlodipine
400
Bisoprolol
Intensity (mV)
500
300
200
100
0
0
10
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Bisoprolol
3.7
1.4
6032
Amlodipine
6.3
1.5
6766

07
Purospher STAR LP Ciprofloxacin Eye drops

Column:
Injection:
20 L
Detection:
Shimadzu Prominence, UV 280 nm
Cell:
10 L
Flow Rate:
1.5 mL/min
Mobile Phase (v/v):
1.7 g of tetra-butyl ammonium phosphate in 1L water. Adjust pH to 2.0

with ortho-phosphoric acid. Mix methanol & buffer 1:3 (v/v)
30 C
Diluent:
Water
Standard:
0.14 mg/ml in water
Sample:
12 mg equivalent of ciprofloxacin from eye drop in 100 ml water.
Pressure Drop:
246 Bar (3567 psi)
Ethylenediamne analog
Intensity (mV)
50
40
30
20
10
Ciprofloxacin
Temperature:
0
0
10
15
20
No.
Compound
Time (min)
RRT
Theoretical Plate
Ethylenediamine analog
5.8
0.8
6095
Ciprofloxacin
7.4
1.0
5090

08
Purospher STAR LP Cyanocobalamine Injection

Time
(min)
%A
%B
0.0
80
20
17.0
80
20
17.1
40
60
Column:
Injection:
50 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
30.0
40
60
Mobile Phase (v/v):
A: 2.72 g of KH2PO4 in 800 ml water. Add 3.76 g hexane
30.1
80
20
40.0
80
20
sulfonic acid sodium salt & dilute to 1000 ml with water.

B: Methanol
Gradient Programme:
See table
Temperature:
40 C
Diluent:
Water
Standard:
Weigh 40 mg cyanocobalamine and dilute to 100 ml with water.

Pipette out 5 ml above solution dilute to 50 ml with water
Sample:
Mix 10 injection ampoules. Dilute 3 ml of this solution to 25 ml with water.
Pressure Drop:
183 Bar (2654 psi)
Cyanocobalamine
140
Intensity (mV)
120
100
80
60
40
20
0
0
16
24
32
40

No.
1
Compound
Cyanocobalamine
Time (min)
Tailing Factor
Theoretical Plate
14.5
1.1
6703

09
Purospher STAR LP Fenofibrate Tablet

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
136 mg potassium di-hydrogen phosphate in 1 L water. Adjust pH to 2.9 with

ortho-phosphoric acid. Mix methanol and buffer 4:1 (v/v)
Temperature:
30 C
Diluent:
Mobile phase
Standard:
67 ppm of fenofibrate in mobile phase.
Sample:
Crush 20 capsules. Weigh equivalent to 67 mg of fenofibrate in 100 ml volumetric

flask. Add 80 ml of mobile phase. Sonicate for 25 min. Dilute 10 ml of the
solution to 100 ml with mobile phase and filter through 0.45 m filter paper.
Pressure Drop:
83 Bar (1203 psi)
Fenofibrate
Intensity (mV)
250
200
150
100
50
0
0
10
15
20
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Fenofibrate
9.2
1.0
8260

10
Purospher STAR LP Folic acid tablet

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
9.38 g of sodium perchlorate & 0.75 g of KH2PO4 in 900 ml water

Adjust pH to 7.2 with 0.1 M KOH & dilute to 1000 ml with water
Mix Buffer and Methanol 800:135 (v/v).
Temperature:
30 C
Diluent:
0.57 % K2HPO4 and methanol 800:135 (v/v)
Sample:
Weigh equivalent to average weight of tablet in 100 ml vol. flask

Add 60 ml diluent. Sonicate for 20 min. Dilute with same solvent &
Filter through 0.45 m filter paper.
Pressure Drop:
130 Bar (1885 psi)
Folic Acid
350
Intensity (mV)
300
250
200
150
100
50
0
0
10

No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Folic acid
3.9
1.2
5713

11
Purospher STAR LP Hydrochlorothiazide & Ramipril tablet

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
5.6 g of sodium perchlorate in 500 ml water. pH adjust to 2.5 with ortho-phosphoric acid
Mix Buffer and Acetonitrile 60:40 (v/v)
Temperature:
35 C
Diluent:
Mobile phase
Standard:
Weigh 10 mg of Hydrochlorothiazide & Ramipril separately and dilute to 100 ml

with Acetonitrile. 5 ml of each solution mixed & dilute to 25 ml with mobile phase.
Pressure Drop:
46 Bar (667 psi)
Intensity (mV)
300
250
200
150
100
Ramipril
Hydrochlorothiazide
350
50
0
0
12
15
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Hydrochlorothiazide
2.8
1.1
3407
Ramipril
10.5
1.0
8524

12
Purospher STAR LP Ivermectin Injection

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.5 mL/min
Mobile Phase (v/v):
Mixture of Acetonitrile:Methanol:Water 106:55:39 (v/v/v)
Temperature:
Ambient
Diluent:
Methanol
Sample:
0.4 mg/ml in diluent
Pressure Drop:
118 Bar (1711 psi)
H1B1a
80
60
H1B1b
Intensity (mV)
100
40
20
0
0
10
20
30
40
50
No.
Compound
Time (min)
Tailing Factor
H1B1b
28.0
1.1
H1B1a
38.0
1.1

Resolution
7.9
13
Purospher STAR LP Levofloxacin Tablet

Column:
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
0.05 M Citric acid : 1M Ammonium acetate: Acetonitrile 84:1:15 (v/v/v)
Temperature:
25 C
Diluent:
Water
Sample:
Crush 20 tablets. Take the weight of tablet equivalent to 100 mg Levofloxacin. Dissolve
In 100 ml 0.1N HCl. Dilute 5 ml to 50 ml with water. Filter through 0.45m filter.
Pressure Drop:
130 Bar (1885 psi)
Levofloxacin
700
Intensity (mV)
600
500
400
300
200
100
0
0
10
15
20
25
30
35
40
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Levofloxacin
12.4
1.4
9691

14
Purospher STAR LP Mecobalamine Injection

Column:
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
0.6 mL/min
Mobile Phase (v/v):
2.72 g of KH2PO4 & 3.76 g hexane sulfonate in 1 L water.

pH adjust to 3.5 with ortho-phosphoric acid. Mix Buffer & Acetonitrile 80:20 (v/v)
Temperature:
40 C
Diluent:
Mobile phase
Standard:
25 mg of mecobalamine in 100 ml water. 5 ml of above solution & 50 mg of

benzyl alcohol diluted to 50 ml with water.
Sample:
5 ml of the smaple diluted to 100 ml with water
Precautions:
Sample preparation do be done in dark. Use amber coloured flask & vial.
Pressure Drop:
54 Bar (783 psi)
Benzyl Alcohol
80
Mecobalamine
Intensity (mV)
100
60
40
20
0
0
10
20
30
40

No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Mecobalamine
11.7
1.1
9526
Benzyl Alcohol
17.0
1.1
17457

15
Purospher STAR LP Metformin and Pioglitazone Tablet

Column:
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
1.2 mL/min
Mobile Phase (v/v):
A: 1.38 g NaH2PO4 in 1800 ml water. Add 3.484 g of Sodium pentane sulfonate

pH adjust to 3.0 with ortho-phosphoric acid & dilute to 2000 ml with water.
B: Acetonitrile
Gradient Programme:
See table
Temperature:
30 C
Sample Cooler:
4 C
Diluent:
Mobile phase A and B 60:40 (v/v)
Sample:
200 ppm each in diluent
Pressure Drop:
129 Bar (1870 psi)
800
600
500
400
%A
%B
0.0
90
10
8.0
90
10
12.0
60
40
20.0
60
40
21.0
90
10
30.0
90
10
Pioglitazone
Metformin
Intensity (mV)
700
Time
(min)
300
200
100
0
0
10
15
20
25
30
No.
Compound
Time (min)
Tailing Factor
Metformin
4.9
1.3
Pioglitazone
15.9
1.2

Retention Factor
2.3
16
Purospher STAR LP Nebivolol Tablet

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
6.8 g Potassium di-hydrogen phosphate in 1 L water. Add 2 ml triethyl amine

Adjust the pH to 3.0 with ortho-phosphoric acid.
Mix buffer, Acetonitrile & Methanol 45:25:30 (v/v/v)
Temperature:
Ambient
Diluent:
Mobile phase
Sample:
Powder 20 tablets & weigh equivalent to 5 mg Nebivolol in 100 ml volumetric flask.

Add 70 ml diluent & sonicate for 15 min. Dilute to 100 ml with diluent.
Filter through 0.45 m filter paper
Pressure Drop:
130 Bar (1885 psi)
35
Nebivolol
Intensity (mV)
30
25
20
15
10
5
0
0
10
15
20

No.
1
Compound
Nebivolol
Time (min)
Tailing Factor
Theoretical Plate
9.5
1.1
10574

17
Purospher STAR LP Ofloxacin Tablet

Column:
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
2.0 mL/min
Mobile Phase (v/v):
27.2 g of KH2PO4 in 1 L water. pH adjust to 2.4 with ortho-phosphoric

acid. Mix buffer and acetonitrile 92:8 (v/v)
Temperature:
30 C
Diluent:
Methanol
Standard:
25 ppm in methanol
Sample:
Crush 20 tablets. Weigh 25 mg equivalent of ofloxacin. Disperse in 25 ml methanol.

Sonicate for 25 min. Dilute to 100 ml with methanol & filter through 0.45 m filter paper.
Pressure Drop:
157 Bar (2276 psi)
Ofloxacin
700
Intensity (mV)
600
500
400
300
200
100
0
0
10
20
30
40
Retention time (min)
No.
1
Compound
Ofloxacin
Time (min)
Tailing Factor
Theoretical Plate
20.6
1.3
6747

18
Purospher STAR LP Olanzapine Tablet

Column:
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
3 g of ammonium dihydrogen phosphate in 900 ml water. Add 2 ml of triethyl amine

Dilute to 1L with water. Adjust the pH to 2.5 with ortho-phosphoric acid.
Mix Buffer & Methanol 70:30 (v/v)
Temperature:
30 C
Diluent:
0.01 M HCl
Standard:
10 mg olanzapine in 25 ml acetonitrile. Dilute to 100 ml with 0.01 M HCl
Sample:
Crush 10 tablets. Weigh 10 mg equivalent of olanzapine. Disperse in 25 ml acetonitrile.

Sonicate for 25 min. Dilute to 100 ml with 0.01M HCl & filter through 0.45 m filter paper.
202 Bar (2929 psi)
900
800
700
600
500
400
300
200
100
0
Olanzapine
Intensity (mV)
Pressure Drop:
12
15

No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Olanzapine
4.9
1.2
3842

19
Purospher STAR LP Hydrochlorothiazide and Olmesartan Tablet

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
3.4 g of KH2PO4 in 1 L water. Sonicate & filter through N-66 Nylon membrane filter paper
Mix Buffer & Acetonitrile 55:45 (v/v)
Temperature:
30 C
Diluent:
Buffer:Acetonitrile 50:50 (v/v)
Standard:
26 mg Hydrochlorothiazide & 80 mg Olmesartan diluted to 100 ml with diluent.
Sample:
Weigh equivalent to average weight of the tablet. Dilute to 50 ml with diluent.

Sonicate for 30 min. Dilute 5 ml to 50 ml with diluent & filter through 0.45 m filter paper.
Pressure Drop:
100 Bar (1450 psi)
700
500
400
300
200
Olmesartan
Hydrochlorothiazide
Intensity (mV)
600
100
0
0
10
15
20

No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Hydrochlorothiazide
3.4
1.2
8061
Olmesartan
7.2
1.0
7238

20
Purospher STAR LP Olmesartan and Amlodipine Tablet

Column:
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
7 ml of tri-ethyl amine diluted to 1 L with water. pH adjusted to 3.0 with ortho-phosphoric acid
Mix Buffer & Acetonitrile 60:40 (v/v)
Temperature:
40 C
Diluent:
Mobile phase
Standard:
35 mg of amlodipine in 50 ml mobile phase. 20 mg of Olmesartan in 100 ml

volumetric flask. Add 10 ml of the above solution & dilute to 100 ml with mobile phase
Weigh equivalent to average weight of the tablet in 100 ml volumetric flask.
Add 70 ml of mobile phase & sonicate for 30 min. Dilute to 100 ml with mobile phase.
Filter through 0.45 m filter paper.
Pressure Drop:
98 Bar (1421 psi)
Amlodipine
Intensity (mV)
100
Olmesartan
Sample:
80
60
40
20
0
0
10
15
20

No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Amlodipine
5.7
1.2
10408
Olmesartan
13.3
1.1
12345

21
Purospher STAR LP Olmesartan Medoxemil Tablet

Column:
Injection:
Detection:
20 L
Cell:
10 L
Flow Rate:
1.0 mL/min
Mobile Phase (v/v):
3.4 g of KH2PO4 in 1 L water. Sonicate & filter through N-66

Nylon membrane filter paper. Mix Buffer and Acetonitrile 55:45 (v/v)
Temperature:
30 C
Diluent:
Buffer:Acetonitrile 50:50 (v/v)
Sample:
Crush 20 tablets & weigh equivalent to 20 mg Olmesartan in 100 ml

volumetric flask. Add 70 ml diluent & sonicate for 15 min.
Dilute to 100 ml with diluent. Further 5 ml dilute to 25 ml with the diluent.
Pressure Drop:
100 Bar (1450 psi)
Olmesartan
Intensity (mV)
250
200
150
100
50
0
0
12
15
No.
1
Compound
Olmesartan
Time (min)
Tailing Factor
Theoretical Plate
7.2
1.0
11844

22

Purospher STAR LP - Application Guide

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Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Specification for Purospher STAR LP RP-18 endcapped

High purity silica gel

Purospher STAR LP RP-18e Vs Competition

Back pressure (Bar)

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Bisoprolol, Amlodipine Tablet

Beta blocker &

Ciprofloxacin Eye drops

Folic acid tablet

High blood pressure

Metformin, Pioglitazole Tablet

High blood sugar

Olmesartan, Amlodipine Tablet

Reduce cholesterol &

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Purospher STAR LP Atorvastatin Tablet

Purospher STAR LP RP18e 250x4.6 mm, 5.0 m

Shimadzu 2010 CHT, UV 246 nm

Mobile Phase (v/v):

A: Acetonitrile : Tetrahydrofuran (92.5:7.5 v/v)

B: 5.75 g of ammonium dihydogen phosphate in 1L water.

Acetonitrile: Water 40:60 (v/v)

Crush 20 tablet. Weigh equivalent to 50 mg. of Atorvastatin. Disperse in 10 ml Methanol.

220 Bar to 200 Bar (3190 psi to 2900 psi)

Retention Time (min)

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Purospher STAR LP Beclomethasone, Chlorocresol, Miconazole

Purospher STAR LP RP-18e 250x4.6 mm, 5 m

Retention Time (min)

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Purospher STAR LP Bisoprolol Fumarate and Amlodipine Tablet

Purospher STAR LP RP-18e 250x4.6 mm, 5 m

Shimadzu 2010 CHT, UV 226 nm

Mobile Phase (v/v):

3.45 g Ammonium di-hydrogen phosphate in 1L water. Adjust pH to 3.0

20 mg Bisoprolol & 56 mg amlodipine diluted to 200 ml with diluent.

Weigh equivalent to two tablet. Add mobile phase to 50 ml.

116 Bar (1682 psi)

Retention Time (min)

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Purospher STAR LP Ciprofloxacin Eye drops

Purospher STAR LP RP-18e 250x4.6 mm, 5 m

Shimadzu Prominence, UV 280 nm

Mobile Phase (v/v):

1.7 g of tetra-butyl ammonium phosphate in 1L water. Adjust pH to 2.0

0.14 mg/ml in water

12 mg equivalent of ciprofloxacin from eye drop in 100 ml water.

246 Bar (3567 psi)

Retention Time (min)

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany

Purospher STAR LP Cyanocobalamine Injection

Purospher STAR LP RP-18e 250x4.6 mm, 5 m

Shimadzu 2010 CHT, UV 361 nm

Mobile Phase (v/v):

A: 2.72 g of KH2PO4 in 800 ml water. Add 3.76 g hexane

sulfonic acid sodium salt & dilute to 1000 ml with water.

Weigh 40 mg cyanocobalamine and dilute to 100 ml with water.

Mix 10 injection ampoules. Dilute 3 ml of this solution to 25 ml with water.

183 Bar (2654 psi)

Retention Time (min)

Merck Millipore is a division of Merck KGaA, Darmstadt, Germany