High-performance liquid chromatography linked to inductively coupled plasma mass spectrometry and orthogonal acceleration time-of-flight mass spectrometry for the simultaneous detection and identification of metabolites of 2-bromo-4-trifluoromethyl-[C-13]-acetanilide in rat urine

The use of HPLC coupled to inductively coupled plasma mass spectrometry (IC PMS) and orthogonal acceleration time-of-flight (oa-TOF) for the profiling, identification, and quantification of metabolites in rat urine following t he administration of 2-bromo-4-trifluoromethylacetanilide is described, The metabolites present in the sample were separated by reversed-phase gradien t chromatography with W-diode array detection. The bulk of the eluent (90%) from the UV detector was directed to an ICPMS where bromine-containing met abolites were detected and quantified using ICPMS. The minor portion of the eluent (10%) was taken for oa-TOFMS for identification. By these means, th e metabolites were identified as sulfate and glucuronide conjugates of a ri ng hydroxy-substituted metabolite, a N-sulfate, a N-hydroxylamine glucuroni de, and N- and N-hydroxyglucuronides.