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dc.contributor.CRUESPUniversidade Estadual de Campinaspt_BR
dc.typeArtigo de periódicopt_BR
dc.titleDevelopment of an amperometric biosensor based on glutathione peroxidase immobilized in a carbodiimide matrix for the analysis of reduced glutathione from serumpt_BR
dc.contributor.authorRover, Lpt_BR
dc.contributor.authorKubota, LTpt_BR
dc.contributor.authorHoehr, NFpt_BR
unicamp.authorUniv Estadual Campinas, Fac Ciencias Med, Dept Patol Clin, BR-13083970 Campinas, SP, Brazil Univ Estadual Campinas, Inst Quim, BR-13083970 Campinas, SP, Brazilpt_BR
dc.subjectreduced glutathione determinationpt_BR
dc.subjectserum samplespt_BR
dc.subjectglutathione peroxidasept_BR
dc.subject.wosPerformance Liquid-chromatographypt_BR
dc.subject.wosCarbon-paste Electrodept_BR
dc.subject.wosAntioxidant Enzyme Levelspt_BR
dc.subject.wosCobalt Phthalocyaninept_BR
dc.subject.wosSulfhydryl Compoundspt_BR
dc.description.abstractThe development of an amperometric biosensor for the reduced glutathione determination in serum is described, The biosensor is based on glutathione peroxidase (GSH-Px, EC immobilized onto a pyrolytic graphite-working electrode using carbodiimide as enzymatic condensing reagent. This resulted in an amperometric biosensor with good sensitivity and stability. The reduced glutathione (GSH) was enzymatically converted to glutathione disulfide (GSSG) in the presence of hydroperoxide. which was monitored amperometrically by its electrooxidation at +0.65 V vs. SCE (saturated calomel electrode). Glutathione measurement was carried out by maintaining the ratio between GSH and hydrogen peroxide at 2:1 (25 degreesC), The amperometric response of the biosensor was linearly proportional to the GSH concentration between 1.9 X 10(-5) and 1.4 X 10(-4) mol/l, in 0.1 mol/l phosphate buffer (pH = 7.8), containing 0.1 mol/l KCI and 0.5 mmol/l Na(2)H(2)EDTA, as the supporting electrolyte, In presence of interfering compounds, the recoveries ranged between 97.2% and 110.7%. The biosensor useful lifetime was at least 2 months when it was evaluated after continuous use. Serum samples analyzed by this biosensor showed a good correlation with the results from the spectrophotometric method (Ellman's reagent) used as reference, presenting relative deviations lower than 7.0%. The low apparent Michaelis-Menten constant value. K-M(app) = 1.6 mmol/l. demonstrated that GSH-Px immobilized on pyrolytic graphite exhibited a high affinity to GSH, without loss of enzymatic activity. (C) 2001 Elsevier Science B.V. All rights
dc.relation.ispartofClinica Chimica Actapt_BR
dc.relation.ispartofabbreviationClin. Chim. Actapt_BR
dc.publisherElsevier Science Bvpt_BR
dc.identifier.citationClinica Chimica Acta. Elsevier Science Bv, v. 308, n. 41671, n. 55, n. 67, 2001.pt_BR
dc.sourceWeb of Sciencept_BR
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