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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.

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

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

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

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

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

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

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

05

Purospher STAR LP Beclomethasone, Chlorocresol, Miconazole


Chromatographic Conditions
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

Retention Time (min)

Chromatographic Data
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

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

06

Purospher STAR LP Bisoprolol Fumarate and Amlodipine Tablet


Chromatographic Conditions
Column:

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

Injection:

20 L

Detection:

Shimadzu 2010 CHT, UV 226 nm

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

Retention Time (min)

Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Bisoprolol

3.7

1.4

6032

Amlodipine

6.3

1.5

6766

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

07

Purospher STAR LP Ciprofloxacin Eye drops


Chromatographic Conditions
Column:

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

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

Retention Time (min)

Chromatographic Data
No.

Compound

Time (min)

RRT

Theoretical Plate

Ethylenediamine analog

5.8

0.8

6095

Ciprofloxacin

7.4

1.0

5090

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

08

Purospher STAR LP Cyanocobalamine Injection


Chromatographic Conditions

Time
(min)

%A

%B

0.0

80

20

17.0

80

20

17.1

40

60

Column:

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

Injection:

50 L

Detection:

Shimadzu 2010 CHT, UV 361 nm

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

Retention Time (min)


Chromatographic Data
No.
1

Compound
Cyanocobalamine

Time (min)

Tailing Factor

Theoretical Plate

14.5

1.1

6703

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

09

Purospher STAR LP Fenofibrate Tablet


Chromatographic Conditions
Column:

Purospher STAR LP RP-18e 150x4.6 mm, 5 m

Injection:

20 L

Detection:

Shimadzu Prominence, UV 285nm

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

Retention Time (min)

Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Fenofibrate

9.2

1.0

8260

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

10

Purospher STAR LP Folic acid tablet


Chromatographic Conditions
Column:

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

Injection:

20 L

Detection:

Shimadzu 2010 CHT, UV 277 nm

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

Retention Time (min)


Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Folic acid

3.9

1.2

5713

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

11

Purospher STAR LP Hydrochlorothiazide & Ramipril tablet


Chromatographic Conditions
Column:

Purospher STAR LP RP-18e 150x4.6 mm, 5 m

Injection:

20 L

Detection:

Shimadzu Prominence, UV 210nm

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

Retention Time (min)

Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Hydrochlorothiazide

2.8

1.1

3407

Ramipril

10.5

1.0

8524

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

12

Purospher STAR LP Ivermectin Injection


Chromatographic Conditions
Column:

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

Injection:

20 L

Detection:

Shimadzu 2010 CHT, UV 245 nm

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

Retention Time (min)

Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

H1B1b

28.0

1.1

H1B1a

38.0

1.1

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

Resolution
7.9

13

Purospher STAR LP Levofloxacin Tablet


Chromatographic Conditions
Column:

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

Injection:

10 L

Detection:

Shimadzu 2010 CHT, UV 293 nm

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

Retention Time (min)

Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Levofloxacin

12.4

1.4

9691

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

14

Purospher STAR LP Mecobalamine Injection


Chromatographic Conditions
Column:

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

Injection:

10 L

Detection:

Shimadzu Prominence, UV 266nm

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

Retention Time (min)


Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Mecobalamine

11.7

1.1

9526

Benzyl Alcohol

17.0

1.1

17457

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

15

Purospher STAR LP Metformin and Pioglitazone Tablet


Chromatographic Conditions
Column:

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

Injection:

10 L

Detection:

Shimadzu Prominence, UV 230 nm

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

Retention Time (min)

Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Metformin

4.9

1.3

Pioglitazone

15.9

1.2

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

Retention Factor
2.3

16

Purospher STAR LP Nebivolol Tablet


Chromatographic Conditions
Column:

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

Injection:

20 L

Detection:

Shimadzu 2010 CHT, UV 281 nm

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

Retention Time (min)


Chromatographic Data
No.
1

Compound
Nebivolol

Time (min)

Tailing Factor

Theoretical Plate

9.5

1.1

10574

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

17

Purospher STAR LP Ofloxacin Tablet


Chromatographic Conditions
Column:

Purospher STAR LP RP-18e 150x4.6 mm, 5 m

Injection:

10 L

Detection:

Shimadzu Prominence, UV 294 nm

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)

Chromatographic Data
No.
1

Compound
Ofloxacin

Time (min)

Tailing Factor

Theoretical Plate

20.6

1.3

6747

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

18

Purospher STAR LP Olanzapine Tablet


Chromatographic Conditions
Column:

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

Injection:

10 L

Detection:

Shimadzu Prominence, UV 220 nm

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

Retention Time (min)


Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Olanzapine

4.9

1.2

3842

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

19

Purospher STAR LP Hydrochlorothiazide and Olmesartan Tablet


Chromatographic Conditions
Column:

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

Injection:

20 L

Detection:

Shimadzu 2010 CHT, UV 225 nm

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

Retention Time (min)


Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Hydrochlorothiazide

3.4

1.2

8061

Olmesartan

7.2

1.0

7238

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

20

Purospher STAR LP Olmesartan and Amlodipine Tablet


Chromatographic Conditions
Column:

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

Injection:

20 L

Detection:

Shimadzu Prominence, UV 237 nm

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

Retention Time (min)


Chromatographic Data
No.

Compound

Time (min)

Tailing Factor

Theoretical Plate

Amlodipine

5.7

1.2

10408

Olmesartan

13.3

1.1

12345

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

21

Purospher STAR LP Olmesartan Medoxemil Tablet


Chromatographic Conditions
Column:

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

Injection:
Detection:

20 L
Shimadzu 2010 CHT, UV 225 nm

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

Retention Time (min)

Chromatographic Data
No.
1

Compound
Olmesartan

Time (min)

Tailing Factor

Theoretical Plate

7.2

1.0

11844

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


http://www.merckmillipore.com/chromatography
chromatography@merckgroup.com

22