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Titolo:
High-performance liquid chromatographic method to analyze picomole levels of glutathione, cysteine and cysteinylglycine and its application to pre-cancerous rat livers
Autore:
Ohmori, S; Kawase, T; Higashiura, M; Chisaka, Y; Nakata, K; Yamasaki, Y;
Indirizzi:
Tsuyama Natl Coll Technol, Tsuyama 7088509, Japan Tsuyama Natl Coll Technol Tsuyama Japan 7088509 , Tsuyama 7088509, Japan Okayama Univ, Fac Pharmaceut Sci, Okayama 7008530, Japan Okayama Univ Okayama Japan 7008530 harmaceut Sci, Okayama 7008530, Japan Kobe Womens Coll Seto, Seto 7090863, Japan Kobe Womens Coll Seto Seto Japan 7090863 Coll Seto, Seto 7090863, Japan
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 1, volume: 762, anno: 2001,
pagine: 25 - 32
SICI:
1387-2273(20011005)762:1<25:HLCMTA>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
BIOLOGICAL THIOLS; DISULFIDE; MONOBROMOBIMANE; DERIVATIZATION; HOMOCYSTEINE; PLASMA; BLOOD;
Keywords:
glutathione; cysteine; cysteinylglycine;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
28
Recensione:
Indirizzi per estratti:
Indirizzo: Ohmori, S Tsuyama Natl Coll Technol, Numa 624, Tsuyama 7088509, Japan Tsuyama Natl Coll Technol Numa 624 Tsuyama Japan 7088509 Japan
Citazione:
S. Ohmori et al., "High-performance liquid chromatographic method to analyze picomole levels of glutathione, cysteine and cysteinylglycine and its application to pre-cancerous rat livers", J CHROMAT B, 762(1), 2001, pp. 25-32

Abstract

A HPLC-based method for quantifying glutathione, cysteine and cysteine-containing peptide is described. N-(2,4-Dinitrophenylaminoethyl)maleimide was synthesized from ethylenediamine, 2,4-dinitrochlorobenzene and maleic anhydride. The maleimide was reacted at 40 degreesC and pH 5.8 for 10 min with thiol compounds such as glutathione or cysteine. An aliquot of the reaction mixtures was applied to a reversed-phase column (4.6 X 150 mm) of HPLC. When glutathione and cysteine were simultaneously assayed, the column was eluted with a gradient of acetonitrile in potassium phosphate (pH 7.0). The derivatives were monitored at 350 nm. Good liner relationships existed betweenpeak area and concentration of glutathione or cysteine from 10 pmol to 2 nmol. The recovery tests from rat liver homogenate were 99.7 +/-2.2% for glutathione and 104.9 +/-3.8% for cysteine. By this method gamma -glutamylcysteine, cysteinylglycine, and homocysteine could be also quantified. The determination limits of glutathione, cysteine and other thiol compounds were 5 pmol. The method is simple: a sample solution is mixed with the labelling reagent and an aliquot of the reaction mixture is applied to a standard HPLC. The hepatic levels of cysteine and glutathione in pre-cancerous rats weredetermined by this method. The cysteine level in pre-cancerous livers was extremely elevated in comparison to that of the control groups, while no difference was observed in the glutathione contents between the pre-cancerousand control groups. (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 06/04/20 alle ore 08:46:15