HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY METHOD FOR THE DETERMINATION OF PROTEIN GLUTATHIONE MIXED DISULFIDE

We have developed an efficient and sensitive method for the quantifica tion of picomole quantities of protein-glutathione and protein-cystein e mixed disulfides in ocular lens. Performic acid oxidation releases g lutathione and cysteine from the protein-mixed disulfides as glutathio ne sulfonic acid and cysteic acid, respectively. The sulfonic acids ar e derivatized with phenylisothiocyanate, and the phenylthiocarbamyl de rivatives are separated by HPLC using a reversed-phase ODS column, equ ilibrated with 0.14 M sodium acetate, containing 0.1% triethylamine an d 6% acetonitrile. (C) 1994 Academic Press, Inc.