Validation and Optimization of a Simple RP-HPLC Method for Determination of Trimetazidine in Human Serum and its Application in a Pharmacokinetic Study with Healthy Bangladeshi Male Volunteers

Abstract

Trimetazidine is an effective and well-tolerated antianginal drug. In the present study, a simple, sensitive
and specific liquid chromatography (HPLC) method with UV detection was developed and validated for the
quantification of trimetazidine in human serum samples using caffeine as internal standard. Protein precipitation
method with methanol was employed in the extraction of trimetazidine and caffeine from biological matrix. The
chromatographic separation was accomplished on Xterra C18 Column with a mobile phase consisting 0.01 M
potassium dihydrogen phosphate buffer (pH 4.16 ± 0.01 adjusted with orthophosphoric acid, with a solvent system of
triethanolamine and acetonitrile (90:10) at a flow rate of 1.0 ml/min. The chromatogram was monitored at a
wavelength of 207 nm. The method was validated over a linear concentration range of 5-200 ng/ml and limit of
quantification (LOQ) was 5.0 ng/ml with a coefficient of correlation (r2) ≥ 0.996. The intra-day and inter-day
precision expressed as relative standard deviation was 3.40%-11.63% and 1.30%-10.21%, respectively. The average
recovery of trimetazidine from serum was 97.44%. The method was successfully applied to a pharmacokinetic study
after oral administration of modified release trimetazidine hydrochloride tablet (35 mg) in healthy Bangladeshi
volunteers