RP-HPLC, Quality by Design (QbD), Azelnidipine, Metoprolol, Analytical method validation

Abstract

The research demonstrates how Quality by Design concepts and principles as prescribed by ICH guidelines can be used to formulate and evaluate Reverse Phase High Performance Liquid Chromatographic (RP- HPLC) method for Azelnidipine and Metoprolol. Factorial design approach was used with an integration of key RP-HPLC parameters of flow rate pH and percentage of acetonitrile. Optimal analysis conditions were determined through Design Expert software (Version 10.0), employing a Hypersil ODS C18 column (5.0 μ, 25 cm × 4.6 mm), Acetonitrile: Using a flow rate of 0.8 ml/min and buffer (70:30 v/v) pH 4.0, 20µg/mL. Conditions available allowed for good resolution between Metoprolol and Azelnidipine retention times of 4.55 and 14.16 respectively, and favorable system suitability parameters were obtained. At 228 nm, the linearity of the developed method was good in the ranges of 2-12 µg/ml for Azelnidipine and 12.5-75 µg/ml for Metoprolol. The optimized method was validated according to ICH guidelines on analytical method validation. Finally, it was shown to ultimately discriminate Azelnidipine from Metoprolol from the binary case, providing an efficient method for pharmaceutical analysis.