Sensitive determination of trimetazidine in spiked human plasma by HPLC with fluorescence detection after pre-column derivatization with 9-fluorenylmethyl chloroformate

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Sep 1;856(1-2):337-42. doi: 10.1016/j.jchromb.2007.06.036. Epub 2007 Jul 6.

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 degrees 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 x 4.6 mm, i.d., 5 microm) 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%).

MeSH terms

  • Calibration
  • Chromatography, High Pressure Liquid / methods*
  • Fluorenes / chemistry*
  • Humans
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence / methods*
  • Spectrophotometry, Ultraviolet
  • Trimetazidine / blood*
  • Vasodilator Agents / blood*

Substances

  • 9-fluorenylmethoxycarbonyl
  • Fluorenes
  • Vasodilator Agents
  • Trimetazidine