Global metabolic profiling of animal and human tissues via UPLC-MS.

TitleGlobal metabolic profiling of animal and human tissues via UPLC-MS.
Publication TypeJournal Article
Year of Publication2013
AuthorsWant EJ, Masson P, Michopoulos F, Wilson ID, Theodoridis G, Plumb RS, Shockcor J, Loftus N, Holmes E, Nicholson JK
JournalNat Protoc
Volume8
Issue1
Pagination17-32
Date Published2013 Jan
ISSN1750-2799
KeywordsAnimals, Chromatography, Liquid, Humans, Liver, Mass Spectrometry, Metabolome, Multivariate Analysis, Tandem Mass Spectrometry
Abstract

Obtaining comprehensive, untargeted metabolic profiles for complex solid samples, e.g., animal tissues, requires sample preparation and access to information-rich analytical methodologies such as mass spectrometry (MS). Here we describe a practical two-step process for tissue samples that is based on extraction into 'aqueous' and 'organic' phases for polar and nonpolar metabolites. Separation methods such as ultraperformance liquid chromatography (UPLC) in combination with MS are needed to obtain sufficient resolution to create diagnostic metabolic profiles and identify candidate biomarkers. We provide detailed protocols for sample preparation, chromatographic procedures, multivariate analysis and metabolite identification via tandem MS (MS/MS) techniques and high-resolution MS. By using these optimized approaches, analysis of a set of samples using a 96-well plate format would take ~48 h: 1 h for system setup, 8-10 h for sample preparation, 34 h for UPLC-MS analysis and 2-3 h for preliminary/exploratory data processing, representing a robust method for untargeted metabolic screening of tissue samples.

DOI10.1038/nprot.2012.135
Alternate JournalNat Protoc
PubMed ID23222455