Stability Indicating HPLC Method for Simultaneous Assessment of Clopidogrel Bisulfate and Aspirin: Development and Validation
Objectives: The efforts were made to develop a HPLC analytical method for the simultaneous quantitative estimation of aspirin and clopidogrel and aim to identify and estimate the degradation of the drugs under the various stress conditions recommended by ICH guideline. Materials and Methods: The separation of two drugs were done by using LC-20AD liquid chromatograph having SPD-20A UV-vis detector on C18 (4.6 x 150 mm) column which was connected with loop 20μl and with HPLC-Dell system. Mobile phase consisted of acetonitrile: buffer in the ratio of 350:650 v/v. Flow rate was 1.3 ml/min and detection were done at 220 nm. Proportions of solvents and adjustment of mobile phase was carried out by screening. Results: The chromatographic separation of aspirin was noted at 4.299 minutes with average USP tangent of 8332.046 and clopidogrel was at 12.706 min with average of 11886.0397 tangent. A linear correlation was observed between concentration of aspirin and clopidogrel with their dilutions i.e r2 = 0.9986 and 0.9996 respectively. The outcome of study, with limit of detection 0.058 and 0.078 μg/ml and limit of quantification 0.117 and 0.156 μg/ml, for aspirin and clopidogrel revealed the repeatability, reproducibility and robustness of the method. Stability indicating parameters were tested by ICH guideline. Conclusion: It is concluded that the method is linear, reproducible, robust, rugged and stability-indicating for simultaneous calculation. It can be used as a routine quality control method for combined pharmaceutical dosage form and for its kinetic studies.