I am doing a spectophotometric method for quantifying the GSH/GSSG ratio in Ishikawa cell line (endometrial adenocarcinoma) after treatment with NAC and hydrogen peroxyde, using a solution with phosphate/EDTA buffer + DTNB + GR + supernatant for free GSH absorbance (412nm, 25ºC), and a solution with phosphate buffer + NADPH + GR + supernatant for total GSH absorbance (340nm, 30ºC).

I don't know how to translate the values of absorbance to the GSH/GSSG ratio. Does anyone have a sugetion?

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