Bioanalytical Method Development and its Validation by Using RP-HPLC for Determination of Trimetazidine Dihydrochloride in Human Plasma

Trimetazidine dihydrochloride is used in the treatment of chest pain in angina condition. The study was performed to determine the bioavailability/bioequivalence of Trimetazidine in human plasma by utilising caffeine as an internal standard. Percent recoveries were obtained as 95.80 % w/v. for trimetazidine. The Liquid-Liquid extraction (LLE) technique was established for the optimal extraction of Trimetazidine and caffeine from biological matrix. The method involved chromatographic separation on Nucleosil C18 (150×4.6mm, 5μm) with mobile phase water, methanol, acetonitrile, and SHS buffer (60:40:10:03 v/v/v/v) at flow rate of 1.3 mL/min with 25°C as ambient temperature and injection volume of 5μL. Quantitation was obtained using UV detection at 210 nm with retention times of 3.97 and 13.07 minutes for caffeine and Trimetazidine in plasma respectively. By applying statistics and performing recovery studies the method was validated showing a linearity concentration range of 5.04 - 201.60 ng/mL and a lower limit of quantification of 0.5 ng/mL. The method proved to be rapid, accurate, simple and precise, providing useful data for therapeutic drug monitoring in clinical laboratories and pharmacokinetics study.