A Review: Determination of Clopidogrel Bisulphate in biological fluid and
Pharmaceutical Dosage Forms
Pankaj D Savani*1, Vineet Jain2, Hasumati Raj1, Sagar Patel1
1Department
of Quality Assurance,
2Department
of Pharmacognosy,
Shree Dhanvantary Pharmacy College,
Kim, Dist: Surat, Gujarat.
*Corresponding Author:
Pankaj D Savani
Department of Quality Assurance,
Shree Dhanvantary Pharmacy College,
Kim, Dist: Surat
Ph. No. 8980138991
Email: pankajsavani75@gmail.com
No. of Figure: 2
No. of Table: 9
ABSTRACT
Clopidogrel Bisulphate is a mainly use in Anti platelet agent as a ADP receptor blocker and
administration of oral tablet dosage form. Chemically, it is Methyl 2-(2- Chlorophenyl) -2-(6,
7-dihydro thieno [3,2-C] Pyridine-5(4H)-yl) Acetate sulphate. It is used in the treatment of
various cardiovascular diseases like Ischemic stroke as well as in Glomerular inflammation,
Myocardial infarction, Atherosclerosis disease, Unstable Angina. An extensive survey of the
literature published in various analytical and pharmaceutical chemistry related journals has
been conducted and the instrumental analytical methods which were developed and used for
determination as single or combination with other drugs in bulk drugs, formulations and
biological fluids have been reviewed. These reviews of analytical methods including
Spectrophotometric methods like Spectrophotometry, Chromatographic method including
HPLC, HPTLC were reported.
Keywords: Clopidogrel Bisulphate, Analytical methods, Compendial Method, UV
Spectroscopic method, Chromatographic methods.
INTRODUCTION
Clopidogrel Bisulphate is Methyl 2-(2- Chlorophenyl) -2-(6, 7-dihydro thieno [3,2-C]
Pyridine-5(4H)-yl) Acetate sulphate is a novel and unique ADP receptor blockers. [1]
Figure 1: Chemical structure of Clopidogrel Bisulphate
PARAMETERS
Category
Chemical formula
IUPAC Name
Molecular Weight
Characteristic
Solubility
Table 1: Drug Profile [2-3]
DESCRIPTION
Anti-Platelet agent
C16H16ClNO2S·H2SO4
Methyl 2- (2- Chlorophenyl) -2- (6,7- dihydro thieno [3.2-C] Pyridine5 (4H)-yl) Acetate sulphate
419.90 gm/mol
White Crystalline Powder
Soluble in Methanol, Acetonitrile, DMSO and Ethyl acetate
MECHANISM OF ACTION [4]
Clopidogrel Bisulphate acts by inhibiting the ADP receptor on platelet cell membranes. It is
a Prodrug, which requires CYP2C19 for its activation. The drug specifically and irreversibly
inhibits the P2Y12 subtype of ADP receptor, which is important in activation of platelets and
eventual cross-linking by the protein fibrin. In Clopidogrel Bisulphate is fibrinogen
Thienopyridine surface receptor on platelets is inhibits selective irreversible P2Y12 Purinergic
Receptor and also inhibits of ADP as fibrinogen induced platelet aggregation. So that
inhibition of Adenylyl cyclase and reduced the inflammation, Ischemic stroke, Myocardial
infarction, Atherosclerosis disease, Unstable Angina.
Figure 2: Mechanism of action Clopidogrel Bisulphate
PHARMACOLOGICAL ACTION [5]
The platelet is integral to the initiation of thrombosis. Antiplatelet Drugs that affect platelet
function are a fundamental part of primary and secondary management of atherosclerotic
thrombotic disease. That indications for the use of Antiplatelet drugs in the management of
thrombotic diseases include stroke, acute myocardial infarction (AMI), acute coronary
syndrome (ACS), angina, percutaneous coronary intervention (PCI), cardiac surgery, primary
and secondary cardiovascular disease prevention, peripheral vascular disease and thrombotic
disorders such as Atrial fibrillation. The risk versus benefit assessment of continuing or
discontinuing these drugs should be made bearing in mind the proposed surgery and its
inherent risk for bleeding or thrombotic complications as well as decisions relating to
appropriate use of general or some form of regional anesthesia.
MARKETED FORMULATION OF CLOPIDOGREL BISULPHATE
Table 3: Marketed formulation of Clopidogrel Bisulphate
SR
BRAND
COMPANY NAME
FORMULATION
NO.
NAME
1
Clopid
Hetero Drugs Ltd
Tablet
2
Clodrel
Unichem laboratories Ltd.
Tablet
3
Clopivas
Cipla Ltd.
Tablet
4
Clopicard
Okasa pharma Pvt. Ltd.
Tablet
DOSE (mg)
75
75, 150
75, 150
75
ANALYTICAL METHODS
This all are the methods which are used for the determination of Clopidogrel Bisulphate in
marketed formulation and in biological fluids. This all analytical methods are reported which
are seen during the literature survey. This article describes the review on the all reported
analytical methods with specific conditions.
A. COMPENDIALMETHOD:
Clopidogrel bisulphate is high performance liquid chromatographic official method available
in Indian pharmacopeia and united state pharmacopeia.
B. REPORTED METHOD:
1. UV SPECTROSCOPIC METHOD
Spectrophotometric method is versatile and economical particularly for developing countries.
Spectrophotometric method has several advantages such as being easy, less expensive and
less time consuming compared with most of the other methods. A simple, precise and
economical Spectrophotometric method for the estimation of Clopidogrel Bisulphate in
pharmaceutical bulk and tablet dosage form was developed and validated. Various methods
like Q-absorption ratio, Simultaneous equation and Derivative methods are used for
determination of Clopidogrel bisulphate alone or in combination with other drugs in marketed
formulation. Below in table describes the various Spectroscopic methods with the method
description and condition which are reported on review literature.
Compendia
IP[6]
USP[7]
Table 4: Compendial method for Clopidogrel Bisulphate
Method
Column
Mobile phase
Detection Flow rate
wavelength (ml/min)
HPLC
C18 (15 cm x 4.6
Potassium
220 nm
1.0
mm, 5µ)
Phosphate buffer
(pH-7.0) :
Acetonitrile (75:25
v/v)
HPLC
C18 (15 cm x 4.6
Potassium
220 nm
1.0
mm, 5µ)
Phosphate buffer
(pH-7.0) :
Acetonitrile (75:25
v/v)
Table 5: Summary of UV-Spectroscopic method for Clopidogrel Bisulphate
Title
Method
Solvent
λmax
(nm)
Development and Validation of Derivative
Derivative
0.1 N HCl
219
UV-Spectrophotometric Methods for
Spectroscopy
Quantitative Estimation of Clopidogrel in
Bulk and Pharmaceutical Dosage Form
Development and Validation of
Simple UVDistilled
222
Spectrophotometric Method for Clopidogrel Spectroscopy
water
Bisulphate in pure and in film coated tablet
method
dosage form
Stability indicating Spectrophotometric
Stability
Acetonitrile
220
method for Development and Validation of
indicating
Clopidogrel Bisulphate tablet dosage form Spectrophotom
etric method
Ref.
8
9
10
2. CHROMATOGRAPHIC METHOD
Various chromatographic methods are used for the determination of the Clopidogrel
Bisulphate alone or combination with other drugs in various marketed formulation and in
biological fluids like human plasma. Chromatographic methods like Reverse phase High
performance liquid chromatography (RP-HPLC) & High performance thin layer
chromatography (HPTLC) are used for determination of Clopidogrel Bisulphate. Below in
table describes the summary of the various chromatographic methods are used with the
method description.
Table 6: Summary of Chromatographic method for Clopidogrel Bisulphate
Title
Method
Stationary
Mobile
Detection
Ref.
phase
phase
wavelength
(nm)
Validation of assay for
RPDevelosil
Acetonitrile :
225
11
bulk Clopidogrel and for
HPLC
ODS HG-5
phosphate
some tablet forms by
RP C18 (15
(65:35 v/v)
Reverse-Phase Highcm × 4.6 mm,
buffer (pH
Performance liquid
5 µm)
2.85)
chromatography
Development and
RPHypersil
Potassium
220
12
Validation of RP-HPLC
HPLC
BDS C18
dihydrogen
method for the estimation
(250 mm ×
orthophospha
of Clopidogrel Bisulphate
4.6 mm, 5
te buffer:
μm)
Acetonitrile
in a ratio of
(32:68 v/v)
(pH 4.0)
Method Development and
Validation of Clopidogrel
Bisulphate by RP-HPLC
in Bulk and
Pharmaceutical dosage
forms
RPHPLC
Hypersil
BDS C18
column (150
mm x 4.5
mm, 5 µm)
Acetonitrile :
Phosphate
buffer (pH:
3.0) in the
ratio of 60:40
%v/v
224
13
Table 7: Summary of Spectroscopic method for Clopidogrel Bisulphate and its combination
Title
Method
WaveWaveSolvent
Ref
length for
length for
Clopidogrel
other
Bisulphate
drug
nd
Quantitative Analysis for
2 Derivative
254 nm
216 nm
Methanol
14
Clopidogrel Bisulphate
Spectrophotom
and Aspirin by Second
etric Method
Derivative
Spectrophotometric
Method in Pharmaceutical
preparation
Simultaneous estimation
Simple UV246 nm
250.4 nm
Water
15
of Rosuvastatin Calcium
Spectroscopy
and Clopidogrel
method
Bisulphate in combined
pharmaceutical
formulation
Development and
Simple UV270 nm
360 nm
Methanol
16
validation of UVSpectroscopy
Spectrophotometric
method
methods for simultaneous
estimation of Amlodipine
Besylate and Clopidogrel
Bisulphate in bulk and
tablet dosage form
Table 8: Summary of Chromatographic method for Clopidogrel Bisulphate and its
combination
Title
Method Stationary
Mobile phase
Detection
Ref.
phase
wavelength
(nm)
Development and
RPPhenomene
0.3%ortho
226
17
validation of RP-HPLCHPLC
x Luna C18 phosphoric acid
UV method for
column
(v/v) :
simultaneous
(250 mm ×
Acetonitrile
Quantitation of
Clopidogrel bisulphate
and Aspirin in bulk drug
Bio-analytical
determination of
Clopidogrel and
Pantoprazole by RPHPLC method in rat
plasma: Application to
drug interaction study
Simultaneous
Determination of
Clopidogrel and
Pioglitazone by high
performance liquid
chromatography in bulk
drug and dosage forms
4.6 mm, 5
µm)
(40:60 v/v)
RPHPLC
C8 column
(250 mm ×
4.6 mm, 5
μm)
Potassium
dihydrogen
ortho phosphate
buffer (pH -3.0)
: Acetonitrile
(40:60 v/v)
240
18
HPLC
Inertsil
ODS C18
column
(250 mm ×
4.6 mm, 5
μm)
Methanol :
Acetonitrile :
Water (80:10:10
v/v)
230
19
Table 9: Other Chromatographic method for Clopidogrel Bisulphate
Title
Method Stationary
Mobile
Detection
phase
phase
wavelength
A Validated Stability
HPTLC Silica gel 60
Carbon
220 nm
Indicating HPTLC Method for
F254
tetrachlorid
Determination of Aspirin and
e : Acetone
Clopidogrel Bisulphate in
(6:2.4 v/v)
Combined Dosage Form
Determination of Clopidogrel
LCSymmetry
Methanol:
Carboxylic Acid in Human
MS/MS C18 column De-ionized
Plasma by LC-MS/MS
(75 mm ×
water :
4.6 mm,
Formic acid
3.5μm)
(75:25:0.1
v/v/v)
A novel GC-MS for the
GC-MS
META-5X
determination of Clopidogrel
column
bisulphate in bulk and
(30.0 m ×
pharmaceutical dosage forms
0.32 mm ×
0.25 mm)
Ref.
20
21
22
CONCLUSION
The presented review highlights on various analytical methods reported on Clopidogrel
Bisulphate and its combination with other drug. HPLC-HPTLC-UV methods were found to
be most widely used. Various chromatographic conditions are presented in under Table. The
faster time, high sensitivity, specificity and better separation efficiency enable HPLC to be
used frequently for the determination of Clopidogrel Bisulphate in the comparison with the
other methods. Other methods are also useful. In this way various analytical methods for the
estimation of Clopidogrel Bisulphate in bulk or in various matrixes like plasma, alone or in
combination with other drugs is discussed. The presented information is useful for the
researchers especially those involved in the formulation development and quality control of
Clopidogrel Bisulphate in combination with other drug.
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