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

doi:10.1007/978-1-4939-7592-1_20

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 volume › Chapter › Scientific

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 language	English
Title of host publication	Methods in Molecular Biology
Publisher	Springer
Pages	277-282
Number of pages	6
Volume	1730
DOIs	https://doi.org/10.1007/978-1-4939-7592-1_20
Publication status	Published - 1 Jan 2018

Publication series

Name	Methods in Molecular Biology
Volume	1730
ISSN (Print)	1064-3745

Keywords

Gas chromatography
Mass spectrometry
Metabolomics
TCA cycle
TMS-MOX derivatives

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10.1007/978-1-4939-7592-1_20

Cite this

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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.",

keywords = "Gas chromatography, Mass spectrometry, Metabolomics, TCA cycle, TMS-MOX derivatives",

author = "Seifar, {Reza Maleki} and {ten Pierick}, Angela and {van Dam}, {Patricia T.N.}",

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doi = "10.1007/978-1-4939-7592-1_20",

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Analysis of metabolites from the tricarboxylic acid cycle for yeast and bacteria samples using gas chromatography mass spectrometry. / Seifar, Reza Maleki; ten Pierick, Angela; van Dam, Patricia T.N.
Methods in Molecular Biology. Vol. 1730 Springer, 2018. p. 277-282 (Methods in Molecular Biology; Vol. 1730).

Research output: Chapter in Book/Conference proceedings/Edited volume › Chapter › Scientific

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AB - 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.

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

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