HIGH THROUGHPUT ASSAY METHOD FOR THE QUANTITATION OF TRIMETAZIDINE IN HUMAN PLASMA BY LC/MS, WITH SELECTED REACTION MONITORING

Abstract

Following oral ingestion (40mg), trimetazidine was quantified in human plasma, using selected reaction monitoring (SRM). The simplicity of the liquid-liquid extraction procedure, together with the sensitivity and specificity afforded by LC-MS/MS, resulted in a highly sensitive and robust assay method in which 200–250 samples were processed per day. Extraction from plasma into hexane/dichloromethane (1+1) and acetylation with acetic anhydride in the organic phase was followed by a novel back-extraction into water and in situ generated acetic acid. This acetylation procedure was found to be necessary to overcome significant carry-over problems encountered with the underivatised analyte. Separation was achieved on a Phenomenex® Luna C8 reversed phase column and detection by tandem mass spectrometry, with a sample turn-around time of 3 minutes.