Analytical Method Development and Validation for Estimation of Amlodipine Besylate and Chlorthalidone by RP-HPLC Method

Abstract

A new selective, single robust, précised and an accurate combination method would be developed and validated by using an High Performance Liquid Chromatography.. The chromatographic separation was achieved by an column using an Finepak SIL C18T-5 Jasco Packed Column with Mobile Phase ratio Acetonitrile:Methanol:HPLC Grade Water and an 0.2% triethylamine with the ph can be set at 3.00 with the formic acid was selected for the good resolution. The detection wavelength selected for the Amlodipine Besylate and Chlorthalidone by an UV-Spectrophotometer would be an 215nm and an 240nm respectively. Retention Time was found to be 9 minute between the two standard componenets of the drug. This method was validated as per the ICH guidelines by an different parameters like System Suitability, Accuracy, Precision, Robustness and Ruggedness. The linear regression analysis concentration range would be an 50 – 150 μg/mL with the linearity equation of Y = 17129x - 26751(R2 = 0.995). For all the calibration plots showed an good relationship. Therefore, the combination method for the Amlodipine Besylate and Chlorthalidone can be validated by using an RP-HPLC Method.