QbD-based RP-HPLC method development for quantitative computation of phase III composition comprising apixaban and clopidogrel

Abstract

The presented work highlights the use of the quality by design methodology in accordance with International Council on Harmonization (ICH) recommendations for developing an reverse phase high performance liquid chromatographic (RP-HPLC) method for apixaban (APX) and clopidogrel (CLP). An efficient approach based on a factorial design incorporating the essential method parameters of the RP-HPLC method such as flow rate, pH, and proportion of organic phase, is presented. The optimum conditions for analysis were derived by using Design Expert software 10.0 Version, i.e., Hypersil ODS C18 column (5.0 μ, 25 cm × 4.6 mm), methanol and 0.05M potassium dihydrogen phosphate buffer (63.5:36.5%v/v, pH 3) as mobile phase, and 0.8 ml/minute as the flow rate. The derived condition gave excellent resolution between APX and CLP with Rt of 4.89 and 14.35 minutes, respectively, with the best possible system suitability parameters. The developed method presented excellent linearity in the range of 1.25–3.75 μg/ml for APX and 37.5–112.5 μg/ml for CLP at 245 nm. The optimized method was further validated in accordance with ICH guidelines on analytical method validation. Finally, the approach was successful in determining APX and CLP from a binary combination.