Comparison of anion-exchange and ion-modified reversed-phase liquid chromatography for the determination of S-sulfocysteine |
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Authors: | Wanlin Xia Mats Sandberg Stephen G Weber |
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Institution: | aChevron Science Center, University of Pittsburgh, Pittsburgh, PA 15260, USA;bInstitute of Neurobiology, Department of Histology, Göteborgs Universitet, Medicinaregatan 5, Göteborg, Sweden |
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Abstract: | A dual Hg–Au amalgam electrode is used to detect S-sulfocysteine (SSC) in this study. There exist two main components in the acetonitrile (ACN) rat brain extracts, namely, Cl− and GSSG (oxidized glutathione), that are active in our detection system (GSH is not extracted in ACN). Two strong anion-exchange columns from different companies were used to separate the samples under different conditions, but SSC and Cl− were not separated at the optimum detection pH of 5.2. The signal from Cl− was greatly decreased by lowering the potential at the downstream electrode, though it cannot be completely eliminated. While a silver cartridge removed Cl− from micromoles to several millimoles without any negative effect on the SSC signal in aqueous standards, a large negative peak which interferes with SSC detection was unfortunately introduced when a silver cartridge was applied to brain tissue samples. However, SSC and Cl− in the samples are successfully separated by ion-modified reversed-phase LC in acetate buffer at the optimum detection pH (5.2). The separation conditions are 20 mM acetic acid, 2% methanol, 0.5 mM cetyltrimethylammonium p-toluene sulfonate (CTMA) (pH 5.2). Most importantly, the sensitivity of SSC under the optimum separation conditions is not sacrificed. The detection limit is 8 nM (20 μl injected). |
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Keywords: | S-Sulfocysteine Thiol Cysteine |
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