title: Simultaneous quantification of reduced and oxidized glutathione in plasma using a two-dimensional chromatographic system with parallel porous graphitized carbon columns coupled with fluorescence and coulometric electrochemical detection.
abstract: A method for the simultaneous quantification of reduced and oxidized glutathione in human plasma employing a two-dimensional chromatographic system with parallel porous graphitized carbon (PGC) columns coupled with fluorescence (FLD) and coulometric electrochemical detection (ED) has been developed. Post-sampling oxidation of reduced glutathione (GSH) was prevented by derivatizing the -SH group with monobromobimane (MBB) and the glutathione-bimane adduct (GSMB) was detected by FLD. Oxidized glutathione (GSSG) was detected by ED optimized to give lowest possible limits of detection (LOD). The method is fully validated and is currently used for determination of GSH, GSSG and its redox potential in different clinical studies.
authors.affiliation: Institute of Basic Medical Sciences, Department of Nutrition, University of Oslo, P.O. Box 1046 Blindern, N-0316 Oslo, Norway.
authors.forename: Amrit K
authors.initials: AK
authors.lastname: Sakhi
authors.forename: Kjell Magne
authors.initials: KM
authors.lastname: Russnes
authors.forename: Sigbjørn
authors.initials: S
authors.lastname: Smeland
authors.forename: Rune
authors.initials: R
authors.lastname: Blomhoff
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Gundersen
mesh_terms: D000925:Anticoagulants / Q000008:administration & dosage; D016022:Case-Control Studies; D002498:Centrifugation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004563:Electrochemistry / Q000379:methods*; D005260:Female; D005978:Glutathione / Q000097:blood*; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D000925:Anticoagulants; D006108:Graphite; D005978:Glutathione