A versatile LC-MS/MS approach for comprehensive, quantitative analysis of central metabolic pathways. [Mass Spectrometry - Lipidomics & Metabolomics]

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TitleA versatile LC-MS/MS approach for comprehensive, quantitative analysis of central metabolic pathways. [Mass Spectrometry - Lipidomics & Metabolomics]
Publication TypeJournal Article
Year of Publication2018
AuthorsWalvekar A, Rashida Z, Maddali H, Laxman S
JournalWellcome Open Res
Volume3
Pagination122
Date Published2018
ISSN2398-502X
Abstract

Liquid chromatography-mass spectrometry (LC-MS/MS) based approaches are widely used for the identification and quantitation of specific metabolites, and are a preferred approach towards analyzing cellular metabolism. Most methods developed come with specific requirements such as unique columns, ion-pairing reagents and pH conditions, and typically allow measurements in a specific pathway alone. Here, we present a single column-based set of methods for simultaneous coverage of multiple pathways, primarily focusing on central carbon, amino acid, and nucleotide metabolism. We further demonstrate the use of this method for quantitative, stable isotope-based metabolic flux experiments, expanding its use beyond steady-state level measurements of metabolites. The expected kinetics of label accumulation pertinent to the pathway under study are presented with some examples. The methods discussed here are broadly applicable, minimize the need for multiple chromatographic resolution methods, and highlight how simple labeling experiments can be valuable in facilitating a comprehensive understanding of the metabolic state of cells.

DOI10.12688/wellcomeopenres.14832.1
Alternate JournalWellcome Open Res
PubMed ID30345389
PubMed Central IDPMC6171562
Grant ListIA/I/14/2/501523 / / Wellcome Trust-DBT India Alliance / India