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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
180
165
164
157
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
Analysis
Methods
IP
Inflammatory skin
disorder
In-house
Cardiovascular
disease
In-house
Antibiotic
Eye infection
USP/IP
Cyanocobalamine Injection
Vitamin B12
In-house
Fenofibrate Capsule
Fibrate
USP
10
Vitamin B9
IP
11
Hydrochlorothiazide, Ramipril
Tablet
Antihypertensive
In-house
12
Ivermectin Injection
Antiparasitic
River blindness
USP
13
Levofloxacin Tablet
Antibiotic
Bacterial infection
IP
14
Mecobalamine Injection
Vitamin
JP
15
Antidiabetic
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
Angiotensin II
receptor &
Antihypertensive
Cardiovascular
disease
In-house
21
Olmesratan Tablet
Angiotensin II
receptor
Cardiovascular
disease
In-house
22
04
Time
(min)
Flow
(ml/min)
%B
%C
Column:
0.0
1.8
100
Injection:
20 L
20.0
1.8
100
Detection:
35.0
1.5
25
75
Cell:
10 L
40.0
1.5
25
75
Gradient:
See table
55.0
1.5
100
60.0
1.8
100
70.0
1.8
100
25 C
Diluent:
Sample:
Pressure Drop:
1200
Atorvastatin
Intensity (mV)
1000
800
600
400
200
0
0
10
20
30
40
50
Compound
Atorvastatin
Time (min)
Tailing Factor
Theoretical Plate
19.6
1.0
11033
05
Sample:
Pressure Drop:
80
Beclomethasone
120
Miconazole
160
Chlorocresol
Intensity (mV)
200
40
0
0
10
15
20
25
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
06
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Temperature:
30 C
Diluent:
Mobile phase
Standard:
Sample:
Pressure Drop:
600
Amlodipine
400
Bisoprolol
Intensity (mV)
500
300
200
100
0
0
10
Chromatographic Data
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Bisoprolol
3.7
1.4
6032
Amlodipine
6.3
1.5
6766
07
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.5 mL/min
Diluent:
Water
Standard:
Sample:
Pressure Drop:
Ethylenediamne analog
Intensity (mV)
50
40
30
20
10
Ciprofloxacin
Temperature:
0
0
10
15
20
Chromatographic Data
No.
Compound
Time (min)
RRT
Theoretical Plate
Ethylenediamine analog
5.8
0.8
6095
Ciprofloxacin
7.4
1.0
5090
08
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
30.1
80
20
40.0
80
20
See table
Temperature:
40 C
Diluent:
Water
Standard:
Sample:
Pressure Drop:
Cyanocobalamine
140
Intensity (mV)
120
100
80
60
40
20
0
0
16
24
32
40
Compound
Cyanocobalamine
Time (min)
Tailing Factor
Theoretical Plate
14.5
1.1
6703
09
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Temperature:
30 C
Diluent:
Mobile phase
Standard:
Sample:
Pressure Drop:
Fenofibrate
Intensity (mV)
250
200
150
100
50
0
0
10
15
20
Chromatographic Data
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Fenofibrate
9.2
1.0
8260
10
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Temperature:
30 C
Diluent:
Sample:
Pressure Drop:
Folic Acid
350
Intensity (mV)
300
250
200
150
100
50
0
0
10
Compound
Time (min)
Tailing Factor
Theoretical Plate
Folic acid
3.9
1.2
5713
11
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
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:
Pressure Drop:
Intensity (mV)
300
250
200
150
100
Ramipril
Hydrochlorothiazide
350
50
0
0
12
15
Chromatographic Data
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Hydrochlorothiazide
2.8
1.1
3407
Ramipril
10.5
1.0
8524
12
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.5 mL/min
Temperature:
Ambient
Diluent:
Methanol
Sample:
Pressure Drop:
H1B1a
80
60
H1B1b
Intensity (mV)
100
40
20
0
0
10
20
30
40
50
Chromatographic Data
No.
Compound
Time (min)
Tailing Factor
H1B1b
28.0
1.1
H1B1a
38.0
1.1
Resolution
7.9
13
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
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:
Levofloxacin
700
Intensity (mV)
600
500
400
300
200
100
0
0
10
15
20
25
30
35
40
Chromatographic Data
No.
Compound
Time (min)
Tailing Factor
Theoretical Plate
Levofloxacin
12.4
1.4
9691
14
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
0.6 mL/min
Temperature:
40 C
Diluent:
Mobile phase
Standard:
Sample:
Precautions:
Sample preparation do be done in dark. Use amber coloured flask & vial.
Pressure Drop:
Benzyl Alcohol
80
Mecobalamine
Intensity (mV)
100
60
40
20
0
0
10
20
30
40
Compound
Time (min)
Tailing Factor
Theoretical Plate
Mecobalamine
11.7
1.1
9526
Benzyl Alcohol
17.0
1.1
17457
15
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
1.2 mL/min
Gradient Programme:
See table
Temperature:
30 C
Sample Cooler:
4 C
Diluent:
Sample:
Pressure Drop:
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
Chromatographic Data
No.
Compound
Time (min)
Tailing Factor
Metformin
4.9
1.3
Pioglitazone
15.9
1.2
Retention Factor
2.3
16
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Temperature:
Ambient
Diluent:
Mobile phase
Sample:
Pressure Drop:
35
Nebivolol
Intensity (mV)
30
25
20
15
10
5
0
0
10
15
20
Compound
Nebivolol
Time (min)
Tailing Factor
Theoretical Plate
9.5
1.1
10574
17
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
2.0 mL/min
Temperature:
30 C
Diluent:
Methanol
Standard:
25 ppm in methanol
Sample:
Pressure Drop:
Ofloxacin
700
Intensity (mV)
600
500
400
300
200
100
0
0
10
20
30
40
Chromatographic Data
No.
1
Compound
Ofloxacin
Time (min)
Tailing Factor
Theoretical Plate
20.6
1.3
6747
18
Injection:
10 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
Temperature:
30 C
Diluent:
0.01 M HCl
Standard:
Sample:
900
800
700
600
500
400
300
200
100
0
Olanzapine
Intensity (mV)
Pressure Drop:
12
15
Compound
Time (min)
Tailing Factor
Theoretical Plate
Olanzapine
4.9
1.2
3842
19
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
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:
Standard:
Sample:
Pressure Drop:
700
500
400
300
200
Olmesartan
Hydrochlorothiazide
Intensity (mV)
600
100
0
0
10
15
20
Compound
Time (min)
Tailing Factor
Theoretical Plate
Hydrochlorothiazide
3.4
1.2
8061
Olmesartan
7.2
1.0
7238
20
Injection:
20 L
Detection:
Cell:
10 L
Flow Rate:
1.0 mL/min
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:
Pressure Drop:
Amlodipine
Intensity (mV)
100
Olmesartan
Sample:
80
60
40
20
0
0
10
15
20
Compound
Time (min)
Tailing Factor
Theoretical Plate
Amlodipine
5.7
1.2
10408
Olmesartan
13.3
1.1
12345
21
Injection:
Detection:
20 L
Shimadzu 2010 CHT, UV 225 nm
Cell:
10 L
Flow Rate:
1.0 mL/min
Temperature:
30 C
Diluent:
Sample:
Pressure Drop:
Olmesartan
Intensity (mV)
250
200
150
100
50
0
0
12
15
Chromatographic Data
No.
1
Compound
Olmesartan
Time (min)
Tailing Factor
Theoretical Plate
7.2
1.0
11844
22