Book ChapterDOI
Determination of glutathione and glutathione disulfide in biological samples.
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TLDR
5,5'-dithiobis(2-nitrobenzoic acid) (DTNB)-GSSG reductase recycling assay for total glutathione is a specific, sensitive, rapid, and reliable procedure, however, because the method depends on an accurate standard curve, appropriate standards containing the protein precipitating agent are essential.Abstract:
Publisher Summary There are a number of procedures, for example, chemical, enzymatic, and chromatographic for the determination of glutathione (GSH) and glutathione disulfide (GSSG) in biological samples. Enzymatic and chromatographic methods for the determination of glutathione in biological samples are described in this chapter. Because GSH readily oxidizes nonenzymatically and because it is a good substrate of γ-glutamyl transpeptidase, the biological samples are acidified quickly to reduce oxidation of GSH to GSSG and to mixed disulfides, and to inactivate γ-glutamyl transpeptidase. Glutathione oxidizes rapidly at pH values greater than 7. Acid treatment inactivates γ-glutamyl transpeptidase, which catalyzes the reactions that decrease the levels of both GSH and GSSG. The optimum method for treating biological samples depends upon the tissue and the experimental system. The discussed 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB)-GSSG reductase recycling assay for total glutathione is a specific, sensitive, rapid, and reliable procedure. However, because the method depends on an accurate standard curve, appropriate standards containing the protein precipitating agent are essential.read more
Citations
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References
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Journal ArticleDOI
Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues
TL;DR: The use of the foregoing analytical method in the determination of total and oxidized glutathione contents of rat blood, kidney, and liver gave values in good agreement with those obtained by previous investigators.
Journal ArticleDOI
Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine
TL;DR: It is reported here that 2-vinylpyridine is a much better reagent for the derivitization of glutathione, and it is demonstrated that the total glutATHione concentration in mouse plasma is substantially higher than generally reported and that glutathion disulfide constitutes less than 30% of the totalglutathione present.
Journal ArticleDOI
Selective modification of glutathione metabolism
TL;DR: Modifications of glutathione metabolism may be achieved by administration of selective enzyme inhibitors, and also by giving compounds that increase glutATHione synthesis, which are useful in chemotherapy and radiation therapy and in protecting cells against the toxic effects of drugs, other foreign compounds, and oxygen.
Journal ArticleDOI
High-performance liquid chromatography analysis of nanomole levels of glutathione, glutathione disulfide, and related thiols and disulfides.
TL;DR: A rapid and sensitive high-performance liquid chromatography method for determination of nanomole levels of glutathione, glutathiona disulfide, cysteine glutathion-mixed disulfides and 20 related sulfur-containing amino acids or their derivatives has been described.