[Optimization of a spectrophotometry assay of total and oxidized blood glutathione: comparison with a fluorimetric method].
We developed a method for the enzymatic assay of glutathione which is easy to practice, rapid, specific, based on the reaction of the thiol group of glutathione with dithiobis-nitrobenzoic acid after the action of glutathione reductase in the presence of NADPH. This spectrophotometric technique allowed, on the one hand, the determination of total glutathione and on the other hand, that of oxidized glutathione (disulfide), after the blockage of reduced glutathione by 2-vinyl-pyridine. The improvements of the assay of blood glutathione concerned the sample preparation, the reaction sensitivity, thanks to a better definition of the optimal pH and a reduction ot the blockage time by 2-vinyl-pyridine in well defined operating conditions. We compared the performances of our technique with a fluorimetric method. We used our method for the determination of total and oxidized blood glutathione in a control population.
Coutelle
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Département de biochimie médicale et biologie moléculaire, université Bordeaux II, France.
Iron
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A
Higueret
D
D
Cassaigne
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A
D000328:Adult; D005260:Female; D005470:Fluorometry; D005978:Glutathione / Q000097:blood* / Q000378:metabolism; D006801:Humans; D008297:Male; D010084:Oxidation-Reduction; D013050:Spectrometry, Fluorescence / Q000379:methods; D013053:Spectrophotometry / Q000379:methods*
D005978:Glutathione
Optimisation du dosage spectrophotométrique du glutathion sanguin total et oxydé: comparaison avec une méthode fluorimétrique.