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Determination of S ‐nitrosoglutathione in erythrocytes by capillary zone electrophoresis
Author(s) -
Messana Irene,
Rossetti Diana Valeria,
Misiti Francesco,
Vincenzoni Federica,
Tellone Ester,
Giardina Bruno,
Castagnola Massimo
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000501)21:8<1606::aid-elps1606>3.0.co;2-2
Subject(s) - capillary electrophoresis , s nitrosoglutathione , chemistry , chromatography , glutathione , electrophoresis , reproducibility , phosphate buffered saline , biochemistry , enzyme
Abstract A method for separation and quantification of S ‐nitrosoglutathione in red cell extracts by capillary electrophoresis is reported. The method is based on the direct analysis of the metaphosphoric acid erythrocyte extract containing diethylenetriaminepentaacetic acid. Optimization of the method is briefly discussed. Best results in the shortest time were obtained at 25°C, using a coated capillary, 7 kV applied voltage and phosphate sodium 40 mmol/L (pH 2.2) as running buffer. Reproducibility, detection limits, and recoveries of S ‐nitrosoglutathione analyses were checked. The results evidenced that S ‐nitrosoglutathione is formed in erythrocytes treated with S ‐nitrosocysteine, a transnitrosating agent. Under our experimental conditions, the contemporaneous detection and quantification of reduced and oxidized glutathione present in cell extract could also be performed.