Study summary · research use only
Simultaneous analysis of reduced glutathione and glutathione disulfide by capillary zone electrophoresis
Plain-language summary
Paraphrased from the published abstract below — not a verdict on whether anything works.
This report describes modifications to a capillary zone electrophoresis (CZE) method for simultaneous analysis of reduced glutathione (GSH) and glutathione disulfide (GSSG). The authors report that lowering the run-buffer pH from 8.4 to 7.8 improved GSH peak-area reproducibility, and that using red blood cells with glutathione extraction at 5% metaphosphoric acid increased sensitivity and decreased interference. CZE results for clinical samples containing 1000 to 3200 micromol GSH/L RBC and 100 to 400 micromol GSSG/L RBC were highly correlated (r2 >= 0.95) with a commercial assay. The abstract states the method was used for glutathione analysis in both mouse and human red blood cells.
Abstract
This report describes modifications to a CZE method developed by Serru et al. (Clinical Chemistry 2001, 47, 1321-1324) for the simultaneous analysis of reduced glutathione (GSH) and glutathione disulfide (GSSG). Lowering the pH of the run buffer (75 mmol/L boric acid, 25 mmol/L bis-Tris) from pH 8.4 to 7.8 markedly improved GSH peak area reproducibility and allowed multiple samples to be analyzed without changing run buffers due to ion depletion. Sample preparation using red blood cells (RBC) instead of whole blood, combined with glutathione extraction at a lower concentration of metaphosphoric acid (5%), increased assay sensitivity and decreased interference. CZE assay results for clinical samples containing 1000 to 3200 μmol GSH/L RBC and 100 to 400 μmol GSSG/L RBC were highly correlated (r(2) ≥ 0.95) with results obtained using a commercial dithionitrobenze-based glutathione assay. The modified CZE assay has proven useful for the analysis of glutathione in both mouse and human RBC.
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