HPLC analysis of lens GSH and GSSG

Abstract
A sensitive method for the analysis of reduced and oxidized glutathione in bovine lens fractions is described. Samples which have been derivatized with iodoacetic acid and dinitrofluorobenzene have been analyzed by high pressure liquid chromatography. Quantitation of reduced and oxidized glutathione in the sample at nanomole levels has been accomplished using a commercially available Bondapak-amine column. Values for reduced glutathione of approximately 30 umoles/gm wet weight have been found for epithelium preparations while only 4 umoles/gm wet weight were found in the nucleus. Only small amounts of oxidized glutathione were observed, increasing in the inner regions of the lens.