Automated software-guided identification of new buspirone metabolites using capillary LC coupled to ion trap and TOF mass spectrometry

The identification and structure elucidation of drug metabolites is one of the main objectives in in vitro ADME studies. Typical modern methodologies involve incubation of the drug with subcellular fractions to simulate metabolism followed by LC‐MS/MS or LC‐MSn analysis and chemometric approaches for the extraction of the metabolites. The objective of this work was the software‐guided identification and structure elucidation of major and minor buspirone metabolites using capillary LC as a separation technique and ion trap MSn as well as electrospray ionization orthogonal acceleration time‐of‐flight (ESI oaTOF) mass spectrometry as detection techniques. Buspirone mainly underwent hydroxylation, dihydroxylation and N‐oxidation in S9 fractions in the presence of phase I co‐factors and the corresponding glucuronides were detected in the presence of phase II co‐factors. The use of automated ion trap MS/MS data‐dependent acquisition combined with a chemometric tool allowed the detection of five small chromatographic peaks of unexpected metabolites that co‐eluted with the larger chromatographic peaks of expected metabolites. Using automatic assignment of ion trap MS/MS fragments as well as accurate mass measurements from an ESI oaTOF mass spectrometer, possible structures were postulated for these metabolites that were previously not reported in the literature. Copyright © 2006 John Wiley & Sons, Ltd.