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In Vitro Depletion of GSH and Generation of ROS by TNBS To examine the interaction of GSH with TNBS and the subsequent effect on GSH regulation, to an aqueous solution of GSH (1 mm) in Krebs-Henseleit buffer, an equimolar amount of TNBS was added and the level of GSH was examined by HPLC as described previously, which monitors reduced and oxidized GSH (Fariss and Reed, 1987)