Developing and Validation of a High-Performance Liquid Chromatography Method for the Determination of Combined Perindopril, Indapamide and Amlodipine from Pharmaceutical Preparations

Abstract

High Performance Liquid Chromatographic method was developed and validated for the analysis of a pharmaceutical preparation (Triplixam®) consist of Perindopril (PRN), Indapamide (IND) and Amlodipine (AML) which are used for the control of hypertension in patients with high blood pressure. In this developed HPLC method, ACE 5 C18 analytical column (12.5 x 4.6 mm) was used for the analysis of PRN, IND and AML. Acetonitrile and 50 mM phosphate buffer mixture (40:60 v/v) was used as mobile phase. Measurements were obtained at 215 nm wavelength using UV detector. Retention times of PRN, AML and IND were 2.22, 3.48 and 5.05 min. respectively, at selected chromatographical conditions. The developed HPLC method was validated in terms of stability, accuracy, sensitivity, linearity, precision, specificity, ruggedness and robustness in accordance with ICH analytical method. Linear calibration curves were obtained in the range of 1.0-25.0 ppm; 1.0-8.0 ppm and 1.0-40.0 ppm for PRN, IND and AML, respectively. In this HPLC method the limit of detection values (LOD) for PRN, IND and AML were 0.25 ppm, 0.10 ppm and 0.15 ppm while limit of quantification values (LOQ) were 0.50 ppm, 0.25 ppm and 0.40 ppm, respectively.