Liquid chromatography-tandem mass spectrometry assay of reduced and oxidized glutathione and main precursors in mice liver.
A liquid chromatography/tandem mass spectrometry assay of glutathione (GSH), glutathione disulfide (GSSG) and of precursors (gamma-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.
Bouligand
Jérôme
J
UPRES EA3535 Pharmacology and New Cancer Treatments, Institute Gustave Roussy and Paris XI University, Villejuif, France.
Deroussent
Alain
A
Paci
Angelo
A
Morizet
Jackie
J
Vassal
Gilles
G
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005978:Glutathione / Q000032:analysis*; D008099:Liver / Q000737:chemistry*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D010084:Oxidation-Reduction; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D005978:Glutathione