Simultaneous determination of reduced glutathione, glutathione disulphide and glutathione sulphonamide in cells and physiological fluids by isotope dilution liquid chromatography-tandem mass spectrometry

D.T. Harwood, A.J. Kettle, S. Brennan, C.C. Winterbourn

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62 Citations (Web of Science)

Abstract

A stable isotope dilution liquid chromatography tandem mass spectrometry (LC–MS/MS) method wasdeveloped and validated for simultaneously quantifying glutathione (GSH), glutathione disulphide (GSSG)and glutathione sulphonamide (GSA) from biological samples. GSA is a selective product of the reactionof GSH with hypochlorous acid and a potential biomarker ofmyeloperoxidase activity. GSH was detectedas the N-ethylmaleimide alkylated adduct, as formation of this species prevented GSH oxidation occurringduring sample processing. Synthesised stable isotope analogues were used as internal standards toaccurately quantify each target species. The limit of quantification was determined as being 0.1 pmol foreach species and excellent linearity was observed over relevant concentration ranges for biological samples.Relative standard deviationswere
Original languageEnglish
Pages (from-to)3393-3399
JournalJournal of Chromatography B
Volume877
Issue number28
DOIs
Publication statusPublished - 2009

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