DEVELOPMENT AND VALIDATION OF A RP HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF ASPIRINE AND CLOPIDOGREL IN COMBINED TABLET DOSAGE FORMAbstract
In this present study a novel, simple, economical, accurate and precise RP-HPLC method has been developed and validated for simultaneous estimation of aspirine and clopidogrel in combined tablet dosage form. Chromatographic separation was carried out on a Kromasil HD C18 (150 x 4.6 mm, 5µm) column with a mixture of acetonitrile and 0.1% (v/v) orthophosphoric acid aqueous solution in a ratio of 40:60 (v/v) as mobile phase at a flow rate of 1.5 ml/min. Detection was accomplished at 237 nm by employing a DAD (Diode array detector). The retention times were 2.78 and 4.99 min for aspirine and clopidogrel respectively. The method was validated according to ICH guidelines for accuracy, precision, linearity, specificity and sensitivity. Calibration plots were linear over the concentration range 10 – 70 µg/ml for both aspirine and clopidogrel with r2>0.997. Recovery was found to be 99.2 – 99.8% and 99.2 – 99.9% for aspirine and clopidogrel respectively with RSD < 2 for both. There were no interference due to any components of pharmaceutical dosage indicating high specificity of the method. LOD and LOQ value were found to be26.2 ng/ml and 78.2 ng/ml for aspirine and 52.7 ng/ml and 152.1 ng/ml for clopidogrel reflecting high sensitivity of the method. The method was successfully employed for quantitative analysis of commercial product. Validation study revealed that method is specific, rapid, reliable and reproducible, so it can be used for routine quality control study of aspirine and clopidogrel in combined tablet dosage form.