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Titolo:
Determination of biological thiols by high-performance liquid chromatography following derivatization by ThioGlo maleimide reagents
Autore:
Ercal, N; Yang, P; Aykin, N;
Indirizzi:
Univ Missouri, Dept Chem, Rolla, MO 65409 USA Univ Missouri Rolla MO USA 65409 Missouri, Dept Chem, Rolla, MO 65409 USA
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 2, volume: 753, anno: 2001,
pagine: 287 - 292
SICI:
1387-2273(20010405)753:2<287:DOBTBH>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
GLUTATHIONE DISULFIDE; VASCULAR-DISEASE; SAMPLES; N-(1-PYRENYL)MALEIMIDE; HOMOCYSTEINE;
Keywords:
thiols; ThioGlo maleimide;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
18
Recensione:
Indirizzi per estratti:
Indirizzo: Ercal, N Univ Missouri, Dept Chem, 142 Schrenk Hall, Rolla, MO 65409 USA Univ Missouri 142 Schrenk Hall Rolla MO USA 65409 a, MO 65409 USA
Citazione:
N. Ercal et al., "Determination of biological thiols by high-performance liquid chromatography following derivatization by ThioGlo maleimide reagents", J CHROMAT B, 753(2), 2001, pp. 287-292

Abstract

The importance of thiols has stimulated the development of a number of methods for determining glutathione and other biologically significant thiols. Methods that rue currently available, however have some limitations, such as being time consuming and complex. In the present study, a new high-performance liquid chromatography (HPLC) method for determining biological thiols was developed by using 9-Acetoxy-2-(4-(2,5-dihydro-2,5-dioxo-1H-pyrrol-1-yl)phenyl)-3-oxo 3H-naphtho[2,1-b]pyran (ThioGlo(TM)3) as a derivatizing agent. ThioGlo(TM) reacts selectively and rapidly with the thiols to yield fluorescent adducts which can be detected fluorimetrically (lambda (ex)=365 nm, lambda (em)=445 nm). The within-run coefficient of variation for glutathione (GSH) by this method ranges from 1.08 to 2.94% whereas the between-runcoefficient of variation for GSH is 4.31-8.61%. For GSH, the detection limit is around 50 fmol and the GSH derivatives remain stable for 1 month, if kept at 4 degreesC. Results for GSSG and cysteine are also included. The ThioGlo(TM) method is compared to our previous method in which N-(1-pyrenyl)maleimide (NPM) is used to derivatize thiol-containing compounds. The present method offers various advantages over the currently accepted techniques, including speed and sensitivity. (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 01/04/20 alle ore 20:02:01