Development and Validation of a RP-HPLC Method for the Analysis of Trimetazidine Hydrochloride in Bulk Drug and Pharmaceutical Dosage Forms

Abstract

This study presents the development, optimization and validation of a simple HPLC method for the determination of Trimetazidine Dihydrochloride in bulk drug and modified release dosage tablets as Trimetazidine Dihydrochloride modified release tablets is not included in any of the official monographs. Method development was carried out by using different columns to get satisfactory result. Mobile phase composition giving symmetrical peak shape was selected. It was also found that C18 column gives symmetric peaks with high theoretical plates and low tailing factor. Simple, fast, economical, accurate, precise and reproducible HPLC method was developed for the determination of Trimetazidine which works as a myocardial metabolism modifier antianginal agent. A simple and reproducible method was developed for Trimetazidine by Reverse phase high performance liquid chromatography (RP-HPLC). The separation was performed by C18 column at ambient temperature with mobile phase of 10 mM Trietylamine, pH 4.0 adjusted with acetic acid buffer and Acetonitrile at a ratio of 85:15 and flow rate of 1 mL/min. The wavelength for maximum absorbance was selected as 232 nm by spectral scan of the Trimetazidine standard solution in UV-VIS Spectrophotometer. The detection was performed by PDA (Photodiode array detection) detector at 232 nm. . The method which was developed was also validated in complete compliance with the current regulatory guidelines by using well developed analytical method validation techniques and tools which comprises with the analytical method validation parameters like linearity, accuracy, method precision, specificity, system suitability and robustness. The proposed method’s results were found to be satisfactory and are suitable for determination of Trimetazidine for routine quality control of drugs in bulk drug and formulation wavelength. The method was found to be linear with regression coefficient value of 0.999 at concentration of 1.859 – 55.763 μg/mL of Trimetazidine. The range was determined at concentration of 39.67 – 65.25 μg/mL with acceptable accuracy (% Recovery: 93.6 – 101.2) and precision study (% RSD: 0.67 – 2.25) in three replicate analysis at three different concentration level of Trimetazidine sample solution. The method showed good precision with %RSD value of 0.16 – 0.64 for repeatability or intraday precision study and %RSD value of 0.20 – 2.02 for intermediate precision or interday precision study. Accuracy was checked for replicate analysis of three concentration level by standard addition method. % recovery value obtained was 97.40 – 99.42 % with % RSD of 0.08 – 0.78. LOD (Limit of Detection) and LOQ (Limit of Quantification) were found to be 0.307 μg/mL and 1.021 μg/mL respectively. Specificity was checked by comparing the peaks of standard and sample with excipients for presence of any interference. The developed method was found to be robust with pH variation of 3.95 – 4.05 and column temperature variation of 20 °C – 30 °C. System suitability parameters were checked for acceptance limit in terms of asymmetry, theoretical plates, capacity factor and relative standard deviation (%) of the six replicate injection in 10 µL value of standard solution at specific concentration. which is useful for the routine determination of Trimetazidine in bulk drug and in its pharmaceutical dosage form. Further study on degradation pathway of the compound will provide idea about the stability indicating nature of the method to discriminate the active constituent, Trimetazidine from its related impurities and degradants. Keywords: Trimetazidine Dihydrochloride, Myocardial metabolism modifier, HPLC, Photodiode array detection, Method validation.