Quantitative metabolomics using ID-MS

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1191; p. 91
• Main Authors: Wahl, S Aljoscha, Seifar, Reza Maleki, Ten Pierick, Angela, Ras, Cor, van Dam, Jan C, Heijnen, Joseph J, van Gulik, Walter M
• Format: Journal Article
• Language: English
• Published: United States 2014
• Subjects: Carbon Isotopes - metabolism; Chromatography, Liquid - methods; Gas Chromatography-Mass Spectrometry - methods; Mass Spectrometry - methods; Metabolic Flux Analysis - methods; Metabolomics - methods; Metabolomics - statistics & numerical data
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-1170-7_6

Quantitative intracellular metabolite measurements are essential for systems biology and modeling of cellular metabolism. The MS-based quantification is error prone because (1) several sampling processing steps have to be performed, (2) the sample contains a complex mixture of partly compounds with the same mass and similar retention time, and (3) especially salts influence the ionization efficiency. Therefore internal standards are required, best for each measured compound. The use of labeled biomass, (13)C extract, is a valuable tool, reducing the standard deviations of intracellular concentration measurements significantly (especially regarding technical reproducibility). Using different platforms, i.e., LC-MS and GC-MS, a large number of different metabolites can be quantified (currently about 110).