title: Interference of plasmatic reduced glutathione and hemolysis on glutathione disulfide levels in human blood.
abstract: The blood reduced glutathione (GSH)/GSH disulfide (GSSG) ratio is an index of the oxidant/antioxidant balance of the whole body. Nevertheless, data indicating GSH and GSSG physiological levels are still widely divergent, especially those on GSSG, probably due to its low concentration. Standardization in methodological protocols and sample manipulation could help to minimize these discrepancies. Therefore, we have investigated how plasma reduced GSH, which is rapidly oxidized after blood withdrawal, could alter the blood GSSG measurement if the sample is not suitably processed. We have observed that an increase in plasma GSH concentration, due to red blood cell hemolysis, is responsible for a significant overestimation of blood GSSG level. Our results show that, before performing blood GSSG determination, thiols have to be rapidly blocked, to avoid possible pitfalls in GSSG measurement, in particular when hemolysis is present.
authors.affiliation: Department of Neuroscience, Pharmacology Section, University of Siena, via A. Moro 4, I-53100, Siena, Italy. giustarini@unisi.it
authors.forename: Daniela
authors.initials: D
authors.lastname: Giustarini
authors.forename: Isabella
authors.initials: I
authors.lastname: Dalle-Donne
authors.forename: Roberto
authors.initials: R
authors.lastname: Colombo
authors.forename: Aldo
authors.initials: A
authors.lastname: Milzani
authors.forename: Ranieri
authors.initials: R
authors.lastname: Rossi
mesh_terms: D000818:Animals; D005978:Glutathione / Q000097:blood* / Q000494:pharmacology; D019803:Glutathione Disulfide / Q000097:blood* / Q000187:drug effects; D006454:Hemoglobins / Q000378:metabolism*; D006461:Hemolysis; D006801:Humans; D051381:Rats
chemical_list: D006454:Hemoglobins; D005978:Glutathione; D019803:Glutathione Disulfide