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.
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.
Sakhi
Amrit K
AK
Institute of Basic Medical Sciences, Department of Nutrition, University of Oslo, P.O. Box 1046 Blindern, N-0316 Oslo, Norway.
Russnes
Kjell Magne
KM
Smeland
Sigbjørn
S
Blomhoff
Rune
R
Gundersen
Thomas E
TE
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*
D000925:Anticoagulants; D006108:Graphite; D005978:Glutathione