High-throughput quantitative and qualitative analysis of microsomal incubations by cocktail analysis with an ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometer system. (April 2015)

• Record Type: Journal Article
• Authors: Backfisch, Gisela; Reder-Hilz, Beate; Hoeckels-Messemer, Juliane; Angstenberger, Jonas; Sydor, Jens; Laplanche, Loic; Schulz, Michael
• Is Part Of: Bioanalysis. Volume 7:Number 6(2015)
• Page Start: 671
• Page End: 683
• Publication Date: 2015-04
• Subjects: Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio.14.314
• Languages: English
• ISSNs: 1757-6180

Abstract

Background: 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.Materials & methods: 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.Results: 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.Conclusion: 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.