The complete targeted profile of the organic acid intermediates of the citric acid cycle using a single stable isotope dilution analysis, sodium borodeuteride reduction and selected ion monitoring GC/MS

Metabolomics : Official Journal of the Metabolomic Society
O A MamerDaina Avizonis

Abstract

The quantitative profiling of the organic acid intermediates of the citric acid cycle (CAC) presents a challenge due to the lack of commercially available internal standards for all of the organic acid intermediates. We developed an analytical method that enables the quantitation of all the organic acids in the CAC in a single stable isotope dilution GC/MS analysis with deuterium-labeled analogs used as internal standards. The unstable α-keto acids are rapidly reduced with sodium borodeuteride to the corresponding stable α-deutero-α-hydroxy acids and these, along with their unlabeled analogs and other CAC organic acid intermediates, are converted to their tert-butyldimethylsilyl derivatives. Selected ion monitoring is employed with electron ionization. We validated this method by treating an untransformed mouse mammary epithelial cell line with well-known mitochondrial toxins affecting the electron transport chain and ATP synthase, which resulted in profound perturbations of the concentration of CAC intermediates.

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Citations

Sep 4, 2014·Cancer & Metabolism·Sylvia AndrzejewskiJulie St-Pierre
Jun 25, 2014·Metabolites·Simon-Pierre GravelJulie St-Pierre
Feb 26, 2015·Diabetes·Kelli M SasSubramaniam Pennathur
Aug 27, 2015·Journal of Biomolecular Structure & Dynamics·Kalyani Korla
Mar 29, 2014·International Journal of Obesity : Journal of the International Association for the Study of Obesity·E Rodríguez-GallegoJ Joven
May 22, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xiaojian ShaoHongxia Liu
Mar 19, 2014·The Journal of Experimental Medicine·Meghan A Wynosky-DolfiIgor E Brodsky
Sep 6, 2017·International Journal of Molecular Sciences·Qiuan ZhongXue Pan
Sep 15, 2016·Nature Communications·Mélissa CarbonneauFrédérick A Mallette
Jun 25, 2020·Metabolomics : Official Journal of the Metabolomic Society·Gunter van der Walt, Roan Louw
Mar 5, 2019·Metabolomics : Official Journal of the Metabolomic Society·David J BealeDaniel A Dias
Dec 11, 2019·Genes·Christine GermeysLudo Van Den Bosch
May 28, 2019·Metabolomics : Official Journal of the Metabolomic Society·Jordan WilkinsEugenia Trushina

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Methods Mentioned

BETA
electrophoresis

Software Mentioned

MSD Chemstation
Isoform ©
GraphPad Prism
GraphPad

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