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/Conference proceedings/Edited volumeChapterScientific

    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 publicationMethods in Molecular Biology
    PublisherSpringer
    Pages277-282
    Number of pages6
    Volume1730
    DOIs
    Publication statusPublished - 1 Jan 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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