An HPLC Determination of Trimetazidine in Human Plasma Using Liquid‐Liquid Extraction for Sample Clean‐Up

Abstract 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 207 nm. Analytes were extracted from plasma samples that previously were mixed with 300 µL saturated K2CO3 solution into an ethyl acetate phase. HPLC separation was accomplished at 40°C on a reversed‐phase column using a mobile phase, 15% acetonitrile in 50 mM potassium dihydrogen phosphate and phosphoric acid (pH=4.0), at a flow‐rate of 1.0 mL/min. The linear range of the method was between 10‐ and 150 ng/mL of tirmetazidine dihydrochloride in human plasma and the quantification limit was 10 ng/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.