Abstract
A high-performance liquid chromatographic method for the determination of trimetazidine dihydrochloride (TMZ) in spiked human plasma is described. The method is based on the pre-column derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) using the fluorimetric detection technique. Fluoxetine HCl (FLX) was used as internal standard. Both, TMZ and FLX were completely derivatized after heating at 50
°C for 20
min in borate buffer pH 8.0. Samples were analyzed by high performance liquid chromatography (HPLC) using Zorbax-TMS column (250
mm
×
4.6
mm, i.d., 5
μm) and mobile phase consist of acetonitrile, methanol and 20
mM sodium acetate pH 4.7 (44:6:50; v/v/v). Fluorescence detector (FLD) was adjusted at excitation and emission wavelengths; 265 and 311
nm, respectively. The linearity of the method was in the range of 4.5–200
ng/ml. Limits of detection (LOD) and quantification (LOQ) were 1.5 and 4.5
ng/ml, respectively. Trimetazidine recovery was 96.5
±
1.3% (
n
=
6; RSD
=
2.1%).