2.17. Determination of Oxidized Glutathione (GSSG) Concentration

The concentration of the oxidized glutathione-GSSG was measured according to a method previously described by Rahman et al., (2006) [28]. The reaction assay contained: 100 μL of liver homogenate (diluted) mixed with 2 μL of 2-vinylpyridine. The assay was incubated with stirring for 60 min, and then 6 μL of triethanolamine, 60 μL DTNB (2 mg/3 mL), 60 μL GR (10 U/3 mL), and 60 μL NADPH (2 mg/3 mL) were added to the reaction assay. The changes in absorbance were recorded at 412 nm. The concentration of the oxidized glutathione has been expressed in nM/mg protein and obtained from the standard curve of GSSG.