Ultrahigh performance liquid chromatography-tandem mass spectrometry method for fast and robust quantification of anionic and aromatic metabolites

J.M. Buescher, S. Moco, U. Sauer, N. Zamboni

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Quantification of metabolites is of pivotal relevance in biology, where it complements more established omics techniques such as transcriptomics and proteomics. Here, we present a 25 min ion-pairing ultrahigh performance liquid chromatography-tandem mass spectrometry method that was developed for comprehensive coverage of central metabolism (glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle) and closely related biosynthetic reactions. We demonstrate quantification of 138 compounds, including carboxylic acids, amino acids, sugar phosphates, nucleotides, and functionalized aromatics. Biologically relevant isomers such as sugar phosphates are individually quantified by combining chromatographic separation and fragmentation. The obtained sensitivity and robustness enabled the detection of more than half all tested compounds in each of eight diverse biological samples of 0.5-50 mg dry cell weight. We recommend this method for routine and yet comprehensive quantification of primary metabolites in a wide variety of biological matrices. © 2010 American Chemical Society.
Original languageEnglish
Pages (from-to)4403-4412
JournalAnalytical chemistry
Volume82
Issue number11
DOIs
Publication statusPublished - 1 Jun 2010
Externally publishedYes

Fingerprint Dive into the research topics of 'Ultrahigh performance liquid chromatography-tandem mass spectrometry method for fast and robust quantification of anionic and aromatic metabolites'. Together they form a unique fingerprint.

Cite this