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Analysis of Erythrocyte Thiol by DTNB. Cell supernatant (40 μl) was added to 1 ml of 6 mM DTNB (in 143 mM phosphate buffer, pH 7.5, containing 6.3 mM EDTA). The change in absorbance at 412 nm after 1 min was converted to a value for total thiol using a GSH standard curve. The GSH standard were prepared in a 1:2:2 mixture of PBS: 0.435M acetic acid: 10% SSA (containing 2 mM EDTA) ("RBC Mix").