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....

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Bibliographic Details
Published in:Journal of Chromatography A 2006-02, Vol.1104 (1), p.179-189
Main Authors: Sakhi, Amrit K., Russnes, Kjell Magne, Smeland, Sigbjørn, Blomhoff, Rune, Gundersen, Thomas E.
Format: Article
Language:English
Subjects:
Anticoagulants - administration & dosage
Biological and medical sciences
Case-Control Studies
Centrifugation
Chromatography
Chromatography, High Pressure Liquid - instrumentation
Chromatography, High Pressure Liquid - methods
Electrochemical detection
Electrochemistry - methods
Female
Glutathione
Glutathione - blood
Graphite - chemistry
GSSG
Humans
Hypercarb
Investigative techniques, diagnostic techniques (general aspects)
Male
Medical sciences
Middle Aged
Miscellaneous. Technology
Oxidation-Reduction
Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques
Plasma
Sensitivity and Specificity
Spectrometry, Fluorescence - methods
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Summary: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.
ISSN:0021-9673
DOI:10.1016/j.chroma.2005.11.129