Analysis of metabolites from the tricarboxylic acid cycle for yeast and bacteria samples using gas chromatography mass spectrometry

Reza Maleki Seifar*, Angela ten Pierick, Patricia T.N. van Dam

*Corresponding author for this work

Research output: Chapter in Book / Report / Conference proceedingChapterAcademicpeer-review

Abstract

We here explain step by step the implementation of gas chromatography coupled with tandem mass spectrometry for the quantitative analysis of intracellular metabolites from the tricarboxylic acid (TCA) cycle such as citrate, isocitrate, alpha-ketoglutarate, succinate, malate, and fumarate. Isotope dilution is used to correct for potential metabolite losses during sample processing, matrix effects, incomplete derivatization, and liner contamination. All measurements are performed in selected reaction monitoring (SRM) mode. Standards and samples are first diluted with a fixed volume of a mixture of fully 13C-labeled internal standards and then derivatized to give trimethylsilyl-methoxylamine derivatives prior GC-MS/MS analysis.

Original languageEnglish
Title of host publicationClinical Metabolomics
Subtitle of host publicationmethods and protocols
EditorsMartin Giera
PublisherHumana Press Inc
Pages277-282
Number of pages6
ISBN (Electronic)9781493975921
ISBN (Print)9781493975914
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1730
ISSN (Print)1064-3745

Keywords

  • Gas chromatography
  • Mass spectrometry
  • Metabolomics
  • TCA cycle
  • TMS-MOX derivatives

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