DEVELOPMENT AND VALIDATION OF A RP - HPLC METHOD FOR THE SIMULTANEOUS DETERMINATION OF AZELNIDIPINE AND CHLORTHALIDONE IN PURE AND PHARMACEUTICAL DOSAGE FORM

Analytical Method Development and Validation for Azelnidipine and Chlorthalidone in bulk and Combined Dosage Form by RP-HPLC. New method was established for simultaneous estimation of Azelnidipine and Chlorthalidone by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Azelnidipine and Chlorthalidone by using Inertsil C18 (4.6mm ×250mm, 5µm particle size), flow rate was 1.0 ml/min, mobile phase ratio was (55:45% v/v) Methanol: Phosphate buffer pH 4.8 (pH was adjusted with ortho phosphoricacid), detection wavelength was 282nm. The instrument used was WATERS Alliance 2695 separation module, Software: Empower 2, 996 PDA detector. The retention times were found to be 1.688mins and 3.282mins. The % purity of Azelnidipine and Chlorthalidone was found to be 99.86%. The system suitability parameters for Azelnidipine and Chlorthalidone such as theoretical plates and tailing factor were found to be 7586, 1.69 and 6235 and 1.58, the resolution was found to be 10.85. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study of Azelnidipine and Chlorthalidone was found in concentration range of 100µg-500µg and 30µg-70µg and correlation coefficient (r2) was found to be 0.999 and 0.999, % recovery was found to be 100.112% and 100.16%, %RSD for repeatability was 0.1702 and 0.043 respectively. The precision study was precise, robust, and repeatable. The LOD value was found to be 2.1µg/ml and 1.28µg/ml, and LOQ value was 6.3µg/ml and 3.84µg/ml for Azelnidipine and Chlorthalidone respectively. Hence the suggested RP-HPLC method can be used for routine analysis of Azelnidipine and Chlorthalidone in API and Pharmaceutical dosage form.

Keywords: Azelnidipine and Chlorthalidone, Method Development, Validation, Accuracy, Precision.