Determination of S-Sulfocysteine and S-Sulfoglutathione in Plasma and Red Blood Cells by High Performance Liquid Chromatography

Abstract

A simultaneous fluorometric determination method of S-sulfocysteine and S-sulfoglutathione was established by HPLC using 7-fluoro-4-nitrobenzo-2-oxa-1, 3-diazole (NBD-F) as a prelabeling reagent. Chromatographic conditions were as follows: Column, TSK gel NH₂-60 (150mm×4 mm i.d.); eluent, 0.1M citrate buffer (pH 3.0) containing 10% methanol; flow rate, 1.0ml/min; sample size, 100μl; detection, Ex 475, Em 545nm. Calibration curves for S-sulfocysteine and S-sulfoglutathione were linear over the range of 0.1 to 10nmol. S-Sulfoglutathione was identified as an endogenous compound in rabbit red blood cells by the use of the present method. This method was also applied to the determination of S-sulfocysteine and S-sulfoglutathione in rabbit plasma and red blood cells after injection of sodium bisulfite.