4.2.2. Determination of GSH

Total glutathione was determined according to [10]. Briefly, 10 µL of cytosol obtained from differently treated erythrocytes were added to 2 mL of a reaction mixture containing 1.9 mL of phosphate 0.1 M/ EDTA 0.6 mM buffer, pH 7.4, 30 µL of 5,50 -dithio-bis(2-

nitrobenzoic acid) (DTNB) 10 mM, 100 µL of NADPH 5 mM, and 10 µg glutathione reductase (GR), and analyzed spectrophotometrically at 412 nm. The total decrease of glutathione content after diamide or DDS-NHOH treatment (∆GSH) was expressed as 1-GSH (Diam or DDS-NHOH)/GSH(Basal) [24].
