DETERMINATION OF GLUTATHIONE, CYSTEINE AND N-ACETYLCYSTEINE IN RABBITEYE TISSUES USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AND POSTCOLUMN DERIVATIZATION WITH 5,5'-DITHIOBIS(2-NITROBENZOIC ACID)

A high-performance liquid chromatographic method to determine glutathi one, cysteine and N-acetylcysteine in rabbit retina, vitreous and lens has been developed. The thiols are separated using a 25x0.46-cm octad ecylsilane column with 0.5 M phosphate buffer, pH 3, as mobile phase. The detection, at 412 nm, involves a post-column derivatization with 5 ,5,-dithiobis(2-nitrobenzoic acid) in presence of cationic micelles of hexadecyltrimethylammonium bromide that enhances the sensitivity. The detection limits are 0.21, 0.92 and 0.61 mu mol/g wet sample for glut athione, cysteine and N-acetylcysteine, respectively. (C) 1997 Elsevie r Science B.V.