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Titolo:
Quantitation of reduced and total glutathione at the femtomole level by high-performance liquid chromatography with fluorescence detection: application to red blood cells and cultured fibroblasts
Autore:
Cereser, C; Guichard, J; Drai, J; Bannier, E; Garcia, I; Boget, S; Parvaz, P; Revol, A;
Indirizzi:
Univ Lyon 1, Fac Pharm, Lab Biochim Endocrinienne & MOl, F-69373 Lyon 08, France Univ Lyon 1 Lyon France 08 Endocrinienne & MOl, F-69373 Lyon 08, France Ctr Hosp Lyon Sud, Biochim Lab, UF Lipides Glucides, F-69495 Pierre Benite, France Ctr Hosp Lyon Sud Pierre Benite France F-69495 495 Pierre Benite, France Univ Lyon 1, Fac Med, Hyg Lab, F-69373 Lyon 08, France Univ Lyon 1 Lyon France 08 1, Fac Med, Hyg Lab, F-69373 Lyon 08, France
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 1, volume: 752, anno: 2001,
pagine: 123 - 132
SICI:
1387-2273(20010305)752:1<123:QORATG>2.0.ZU;2-B
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-BOUND GLUTATHIONE; HUMAN-SKIN FIBROBLASTS; OXIDIZED GLUTATHIONE; PRECOLUMN DERIVATIZATION; FLUOROMETRIC DETECTION; HUMAN PLASMA; MITOCHONDRIAL GLUTATHIONE; ERYTHROCYTE GLUTATHIONE; GAMMA-GLUTAMYLCYSTEINE; PARKINSONS-DISEASE;
Keywords:
pre-column derivatization; HPLC; glutathione; o-phthalaldehyde;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
43
Recensione:
Indirizzi per estratti:
Indirizzo: Revol, A Univ Lyon 1, Fac Pharm, Lab Biochim Endocrinienne & MOl, 8 Av Rockefeller,F-69373 Lyon 08, France Univ Lyon 1 8 Av Rockefeller Lyon France 08 9373 Lyon 08, France
Citazione:
C. Cereser et al., "Quantitation of reduced and total glutathione at the femtomole level by high-performance liquid chromatography with fluorescence detection: application to red blood cells and cultured fibroblasts", J CHROMAT B, 752(1), 2001, pp. 123-132

Abstract

A new rapid and highly sensitive HPLC method with ortho-phthalaldehyde (OPA) pre-column derivatization has been developed for determination of reduced glutathione (GSH) and total glutathione (GSHt) in human red blood cells and cultured fibroblasts. OPA derivatives are separated on a reversed-phase HPLC column with an acetonitrile-sodium acetate gradient system and detected fluorimetrically. An internal standard (glutathione ethyl ester) is addedto facilitate quantitation. Total glutathione is determined after reduction of disulfide groups with dithiothreitol; the oxidized glutathione (GSSG) concentration is calculated by subtraction of the GSH level from the GSHt level. The assay shows high sensitivity (50 fmol per injection, the lowest reported), good precision (C.V. <5.0%), an analytical recovery of GSH and GSSG close to 100%, and linearity (r > 0.999). This HPLC technique is very simple and rapid. Its wide applicability and high sensitivity make it a convenient and reliable method for glutathione determination in various biological samples. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/02/20 alle ore 15:10:03