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RESEARCH PRODUCT

[23] Ratio of reduced to oxidized glutathione as indicator of oxidative stress status and DNA damage

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Publisher Summary This chapter discusses the ratio of reduced to oxidized glutathione (GSH) as an indicator of oxidative stress status and DNA damage. Several methods have been proposed for the determination of GSH status in biological samples. Accurate determination of this status is largely dependent on the prevention of GSH autoxidation during sample processing. As the disulfide form (GSSG) is present only in minimal amounts with respect to the reduced form, a small GSH autoxidation during sample processing can give erroneously high GSSG level. The chapter describes high-performance liquid chromatography (HPLC) method for determining GSSG. It also presents a method for glutathione determination and explores the possible relationship between glutathione oxidation and apoptosis in fibroblasts. The resulting glutathione redox ratio (GSSG/GSH) accurately reflects the redox ratio of cells or of subcellular organelles and relationships can be found between glutathione oxidation and other metabolic parameters, such as lactate levels in exercise, oxidized DNA bases in aging, or apoptosis. The methods described in the chapter may be used to determine the glutathione redox ratio in various physiological and pathophysiological situations.