An HPLC determination of Trimetazidine in human plasma using liquid-liquid extraction for sample clean-up

Trimetazidine dihydrochloride has been used as an antianginal drug that possess protective properties against ischemia-induced damage to heart. A simple and sensitive analytical method of trimetazidine dihydrochloride in human plasma by using high performance liquid chromatography (HPLC) was developed. The method employs a liquid-liquid extraction for isolation and sample concentration, followed by reversed-phase liquid chromatography (RPLC) analysis using ultraviolet (UV) detection at 207nm. Analytes were extracted from plasma samples that previously were mixed with 300 mu L saturated K2CO3 solution into an ethyl acetate phase. HPLC separation was accomplished at 40 degrees C on a reversed-phase column using a mobile phase, 15% acetonitrile in 50mM potassium dihydrogen phosphate and phosphoric acid (pH = 4.0), at a 11 flow-rate of 1.0 mL/min. The linear range of the method was between 10- and 150ng/mL of tirmetazidine dihydrochloride in human plasma and the quantification limit was 10ng/mL. The intra- and inter-day relative standard deviation (RSD) were less than 7.6% and the accuracy was in the range of 98-107%. Extraction recoveries ranged from 71.5 to 84.6% and the C.V. values showed between 1.7 and 10.4% in the same concentration range. This method has been sought for carrying out pharmacokinetic studies and for assessing bioavailabiltiy. Such a method would be ideally suitable for pharmacokinetic studies in human volunteers after oral administration of different types of dosages of the drug.