Determination of Clopidogrel Bisulphate from Pharmaceutical Dosage Form by Extractive Ion Pair Complex Colorimetric Method
Rele Rajan V.*
Central Research Laboratory, D.G. Ruparel College, Mahim, Mumbai 400 016.
*Corresponding Author E-mail: drvinraj@gmail.com
ABSTRACT:
In this communication a new colorimetric method is developed for assay of clopidogrel bisulphate in pharmaceutical dosage form. Clopidogrel bisulphate, chemically (+) - (S) -(2-chlorophenyl)- 6,7-dihydrothieno [3,2-c] pyridine- 5(4H)-acetic acid methyl ester sulphate is a potent oral anti-platelet agent often used in the treatment of coronary artery disease, peripheral vascular disease and cerebro vascular disease.
The mechanism of action of clopidogrel is irreversible blockade of the adenosine di-phosphate (ADP) receptor P2Y12 and is important in platelet aggregation, the cross-linking of platelets by fibrin. The blockade of this receptor inhibits platelet aggregation by blocking activation of the glycoprotein IIb/ IIIa pathway. It Literature survey reveals the assay of drug in pharmaceutical formulations by various HPLC [1-5], spectrophotometric [6-11], TLC [12] methods for assay of drug.
Simple, rapid and reliable UV spectrophotometric assay method is suggested for the clopidogrel bisulphate. In the proposed methods optimization and validation of this method are reported.
Fig. 1: Chemical Structure of Clopidogrel
MATERIALS AND METHODS:
A Shimadzu -160 A double beam UV-VISIBLE recording spectrophotometer with pair of 10mm matched quartz cell was used to measure absorbance of solutions. A Shimadzu analytical balance was used.
Cobalt nitrate, Ammonium thiocynate, and chloroform of A.R. grade were used in the study.
Preparation of standard solution and reagents
Stock solution of Clopidogrel bisulphate (100 μg/ml) was prepared in distilled water. From this stock solution working standard (10 μg/ml) was prepared by diluting 10 ml stock solution to 100 ml with distilled water.
EXPERIMENTAL:
Preparation of solutions:
Stock solutions of Clopidogrel bisulphate (100μg/ml) were prepared in distilled water.
From these stock solutions, working standard solutions (10μg/ml) were prepared by diluting 10 ml stock solution to 100 μg/ ml with distilled water.
Preparation of reagents
The 1:2 molar proportions of cobalt nitrate and ammonium thiocynate are mixed to give reagent cobalt thiocynate in distilled water.
Experimental:
The different aliquots of working solution were taken in different separating funnels. In each separating funnel 1.6 ml of reagent (cobalt thiocynate) solution were added with 10 ml of chloroform. The each aliquot with reagent and chloroform were shaken for 10 minutes for thorough mixing. After the separation of the two phases, chloroform layer was taken out. The optical density values of the each chloroform layers was measured at λ max 622 nm with chloroform as reagent blank solution.
Estimation from Tablets:
The finely crushed powder of drug formulation (tablets) equivalent to 100 mg of was used to prepare 100 μg /ml. A 10 μg /ml solution was prepared from above formulation solution of 100 μg /ml. This 10 μg /ml solutions were used for assay of clopidogrel bisulphate respectively. The different aliquots of sample drug solution were used for the determination of drug contents in tablets. The concentration of the drug in the tablets was calculated using calibration curve. Fig. no.1
Fig. no: 2. Calibration curve
The standard addition method was used for recovery experiment. The values result of different parameters are reported in table no1.
Table 1: Values of results of different parameter of drug
|
Parameter |
Values |
|
Detection Wavelength (nm) |
622 |
|
Optical density Limits (µg/ml) |
2-20 |
|
Correlation coefficient(r2) |
0.9998 |
|
Regression equation |
|
|
Slope |
0.0101 |
|
Intercept |
-0.0002 |
RESULT:
The spectrophotometric methods are known for accuracy in quantitative determination of the drug with many pharmaceutical preparations. This suggested assay method is applicable determination of drug from active ingredient as well as formulation. The extractive complexes are formed are very stable. The optimizations of various parameters involved for maximum colour development were studied. The recovery study was statistically studied for the proposed methods. Assay results of recovery studies are given in table no. 2.
Table no 2: Results of recovery studies
|
Amount of Sample Added in (µg/ml) |
Amount of Standard Added in (µg/ml) |
Total Amount Recovered |
Percentage Recovery (%) |
Standard Deviation |
Percentage of relative standard Deviation (C.O.V.) |
|
2 |
0 |
2.015714 |
100.7857 |
0.009759 |
0.484146 |
|
2 |
2 |
4.016 |
100.4 |
0.012329 |
0.306993 |
|
2 |
4 |
6.018571 |
100.3095 |
0.006901 |
0.114656 |
|
2 |
6 |
8.024286 |
100.3036 |
0.012724 |
0.158571 |
|
|
|
|
|
Mean= 0.010428 |
Mean=0.266091 |
The noninterference from the common excipient used was observed from percent recovery obtained by calculation.
DISCUSSION:
The low values of standard deviations indicated good reproduction, and accuracy of such methods. The newly suggested method is simple, precise and reproducible with given range of concentration. The method is directly used for active ingredient as well as formulation of estimation of clopidogrel bisulphate in bulk and pharmaceutical dosage form even at very low concentration in formulation such as tablets.
REFERENCES:
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Received on 04.10.2025 Revised on 24.12.2025 Accepted on 28.01.2026 Published on 27.05.2026 Available online from May 30, 2026 Asian J. Research Chem.2026; 19(3):182-184. DOI: 10.52711/0974-4150.2026.00030 ©A and V Publications All Right Reserved
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