Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Fast and Robust Quantification of Anionic and Aromatic Metabolites

被引：278

作者：

Buescher, Joerg Martin ^{[1
]}

Moco, Sofia ^{[1
]}

Sauer, Uwe ^{[1
]}

Zamboni, Nicola ^{[1
]}

机构：

[1] Swiss Fed Inst Technol, Inst Mol Syst Biol, Zurich, Switzerland

来源：

ANALYTICAL CHEMISTRY | 2010年 / 82卷 / 11期

关键词：

PENTOSE-PHOSPHATE PATHWAY; SACCHAROMYCES-CEREVISIAE; ELECTROSPRAY-IONIZATION; QUANTITATIVE-ANALYSIS; POLAR METABOLITES; METABOLOMICS; EXTRACTS; COMPLEX; YEAST; ACID;

D O I：

10.1021/ac100101d

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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.

引用

页码：4403 / 4412

页数：10

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