AN IMPROVED RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF ASPIRIN, ATORVASTATIN, AND CLOPIDOGREL IN PHARMACEUTICAL FORMULATION USING EXPERIMENTAL DESIGN METHODOLOGY

Abstract

Objective: In this study an improved RP-HPLC method was developed for the simultaneous estimation of Aspirin, Atrovastatin, and Clopidogrel in pharmaceutical dosage form. For improving the separation, an experimental design approach was employed. Methods: Factors–independent variables (Organic modifier, pH of the mobile phase and flow rate) were extracted from the preliminary study and as dependent three responses variables viz. Capacity factor of tR1, resolution between Atorvastatin and internal standard, retention time of tR4 were selected. To improve method development and optimization, Derringer's desirability function was applied to simultaneously optimize the chosen three responses. Results: The procedure allowed deduction of optimal conditions and the predicted optimum was Acetonitrile: Methanol: 0.1% of Triethylamine (52:05:43, v/v/v), pH of the aqueous phase adjusted at to 3.0 with 10 % ortho phosphoric acid, and the separation was achieved within 8 minutes. The method showed good agreement between the experimental data and predictive value throughout the studied parameter space. Conclusion: The optimized assay condition was validated according to International Conference on Harmonisation (ICH) guidelines to confirm specificity, linearity, accuracy and precision. The proposed validated method was successfully applied for the analysis of commercially available dosage form