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An enantiospecific RP-HPLC method was developed and validated for the simultaneous determination of clopidogrel and four related compounds specified as impurities. Experimental design was applied during the method optimization (Full factorial 23 design) and robustness testing (Central Composite Face Centered design). Laboratory mixtures of clopidogrel and its impurities in a concentration ratio of 1: 5.0×10 –4 were used as an investigation matrix. The three independent variables were the acetonitrile content in the mobile phase, pH of the mobile phase, and the column temperature. A Chromatographic Response Function (CRF) was used for estimation of the system response resolution (Rs). Separation was achieved using mobile phase composition of ACN: Buffer solution pH 6.5 (40:60 v/v) at 30 oC. A CHIRAL-AGP 4.0 mm × 100 mm, 5.0 μm particle size column was used. The total time for chromatographic separation was approximately 10.0 min. The method was validated for its selectivity, linearity, precision, accuracy and robustness.

macedonian journal of chemistry and chemical engineering

Experimental design approach for the development and validation of an enantiospecific RP-HPLC method for simultaneous determination of clopidogrel and related compounds