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Determination of oxidized and reduced glutathione, by capillary zone electrophoresis, in Brassica juncea plants treated with copper and cadmium
Author(s) -
Mendoza Jorge,
Soto Paula,
Ahumada Inés,
Garrido Tatiana
Publication year - 2004
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/elps.200305759
Subject(s) - capillary electrophoresis , chemistry , chromatography , electrolyte , detection limit , analyte , calibration curve , glutathione , electrophoresis , cadmium , matrix (chemical analysis) , analytical chemistry (journal) , electrode , biochemistry , organic chemistry , enzyme
Abstract A rapid method of capillary zone electrophoresis is described to determine the oxidized (GSSG) and reduced (GSH) form of glutathione in plant tissue. In order to separate both analytes in a fused‐silica capillary, the pH and composition of the electrolyte solution were optimized. The electrolyte composition was 100 mmol/L, borate 25 mmol/L Tris, and 0.2% w/v metaphosphoric acid (MPA), pH 8.2. Some instrumental conditions used to run the samples were hydrostatic injection for 30 s, 30 kV applied voltage, and UV detection (185 nm) at 25°C. Linearity and useful range obtained for the calibration curves were optimum, with correlation coefficients about 0.999 in the 0–120 μmol/L range. The migration time was highly reproducible, less than 5 min being afforded to run a sample. Electrolyte buffer and samples required a careful pH control for optimal separation of both analytes. This aspect constitutes a critical analytical step when acids are used in the procedure for sample preparation. Simultaneous analysis of GSH and GSSG may provide a useful tool for comparative studies of plants in order to select those species with a potential capacity for detoxification from toxic elements or those appearing promising from phytoremediation for these elements.