Liquid chromatography-tandem mass spectrometry assay of reduced and oxidized glutathione and main precursors in mice liver

Jérôme Bouligand, Alain Deroussent, Angelo Paci, Jackie Morizet, Gilles Vassal

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    Résumé

    A liquid chromatography/tandem mass spectrometry assay of glutathione (GSH), glutathione disulfide (GSSG) and of precursors (γ-glutamyl- cysteine, cysteinyl-glycine, cysteine, cystine, homocysteine and homocystine) was developed to study glutathione synthesis in mice liver. After iodoacetic acid derivatization, the analytes were analyzed using reversed-phase gradient HPLC and detected using multiple reaction monitoring. Linear calibrations were performed over the concentrations range of 100-10,000 ng/mL for the thiol-containing precursors and extended up to 100,000 ng/mL for GSH and GSSG. The method was validated for each compound with inter-day accuracy below 11.9% and with precision below 15%. The method showed low limits of quantitation of 100 ng/mL for each thiol-containing compound and GSSG and of 200 ng/mL for other disulfides.

    langue originaleAnglais
    Pages (de - à)67-74
    Nombre de pages8
    journalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
    Volume832
    Numéro de publication1
    Les DOIs
    étatPublié - 17 févr. 2006

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