Development and Validation of Reversed Phase HPLC Method for Analysis of Ranitidine, Cimetidine, and Acetosal Compounds Using Factorial Design

Abstract

Differences in physicochemical properties between ranitidine, cimetidine, and acetosal such as pKa and log P values ​​can affect the analysis results in the High Performance Liquid Chromatography (HPLC) method. The purpose of this study was to determine the best chromatography system for HPLC that is optimal in separating compounds and ensuring the validation of the method. The research method used laboratory experiments in the development of a reversed phase HPLC system using a factorial design. The variables analyzed were flow rate, pH, and mobile phase buffer concentration. Method validation was carried out with selectivity, linearity, LOD, LOQ, accuracy, and precision parameters. The results showed the best conditions using a 230 nm wavelength UV detector, a flow rate of 0.5 mL/min, a mobile phase of acetonitrile: 0.05 M acetate buffer at pH 5 (20:80 v/v). Based on the analysis of interactions between variables, it showed an influence on all three variables. The validation results of the linearity test for ranitidine, cimetidine, and acetosal were 0.9989; 0.9989; and 0.999, respectively. The LOD value was 2.59 ppm and the LOQ value was 8.64 ppm. The selectivity test on the first peak with the second peak was 1.509 and on the second peak with the third peak was 5.495. The accuracy test with % recovery was 103.863% and % RSD was 1.819%. The method used was proven valid as indicated by the results that met the method validation requirements.