High-throughput quantitative and qualitative analysis of microsomal incubations by cocktail analysis with an ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometer system

• Published in: Bioanalysis Vol. 7; no. 6; pp. 671 - 683
• Main Authors: Backfisch, Gisela, Reder-Hilz, Beate, Hoeckels-Messemer, Juliane, Angstenberger, Jonas, Sydor, Jens, Laplanche, Loic, Schulz, Michael
• Format: Journal Article
• Language: English
• Published: England 01.01.2015
• Subjects: Animals; Chromatography, High Pressure Liquid; Humans; Mass Spectrometry; Metabolomics - methods; Microsomes, Liver - metabolism; Rats; Software
• ISSN: 1757-6180; 1757-6199
• DOI: 10.4155/bio.14.314

Metabolite identification studies are very resource intensive and also are rarely performed in early discovery. Here, we report the validation of an ultraperformance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) platform for generating high-throughput stability data with structure elucidation in a single injection. Tandem mass spectrometry spectra were obtained for quantitative analysis using a generic information-dependent acquisition method from pooled microsomal samples incubated at low compound concentrations. A good correlation was observed between clearance determined using UPLC-HRMS and UPLC-triple-quadrupole analysis. Structural elucidation performed with MassMetaSite™ (Molecular Discovery, Perugia, Italy) software identified 85% of the major metabolites of eight marketed drugs and over 100 internal compounds under these conditions. For the first time, a high-throughput quantitative-qualitative workflow was established using a cocktail approach for sample analysis with UPLC-HRMS in order to enable metabolite identification in early discovery projects.