Simultaneous analysis of reduced glutathione and glutathione disulfide by capillary zone electrophoresis
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...
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| Veröffentlicht in: | Electrophoresis 2014-04, Vol.35 (7), p.967-971 |
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| creator | Hempe, James M. Ory-Ascani, Jeannine |
| description | 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 (r2 ≥ 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. |
| doi_str_mv | 10.1002/elps.201300450 |
| format | Article |
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(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 (r2 ≥ 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.</description><identifier>ISSN: 0173-0835</identifier><identifier>EISSN: 1522-2683</identifier><identifier>DOI: 10.1002/elps.201300450</identifier><identifier>PMID: 24338531</identifier><language>eng</language><publisher>Germany: Blackwell Publishing Ltd</publisher><subject>Animals ; Assaying ; Blood ; Boric acids ; Capillary electrophoresis ; Disulfides ; Electrophoresis ; Electrophoresis, Capillary - methods ; Erythrocytes ; Erythrocytes - chemistry ; Glutathione ; Glutathione - blood ; Glutathione - chemistry ; Glutathione Disulfide - blood ; Glutathione Disulfide - chemistry ; Humans ; Linear Models ; Mice ; Reproducibility ; Reproducibility of Results ; Sensitivity and Specificity</subject><ispartof>Electrophoresis, 2014-04, Vol.35 (7), p.967-971</ispartof><rights>2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4110-64d8d14657733f2ec6e0cf03ad3bcbec11e53acb7b3041b13eb5e5fe4c5c8f563</citedby><cites>FETCH-LOGICAL-c4110-64d8d14657733f2ec6e0cf03ad3bcbec11e53acb7b3041b13eb5e5fe4c5c8f563</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Felps.201300450$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Felps.201300450$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24338531$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hempe, James M.</creatorcontrib><creatorcontrib>Ory-Ascani, Jeannine</creatorcontrib><title>Simultaneous analysis of reduced glutathione and glutathione disulfide by capillary zone electrophoresis</title><title>Electrophoresis</title><addtitle>ELECTROPHORESIS</addtitle><description>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 (r2 ≥ 0.95) with results obtained using a commercial dithionitrobenze‐based glutathione assay. 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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Animals Assaying Blood Boric acids Capillary electrophoresis Disulfides Electrophoresis Electrophoresis, Capillary - methods Erythrocytes Erythrocytes - chemistry Glutathione Glutathione - blood Glutathione - chemistry Glutathione Disulfide - blood Glutathione Disulfide - chemistry Humans Linear Models Mice Reproducibility Reproducibility of Results Sensitivity and Specificity |
| title | Simultaneous analysis of reduced glutathione and glutathione disulfide by capillary zone electrophoresis |
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