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Titolo:
Analysis of free and protein-bound nitrotyrosine in human plasma by a gas chromatography/mass spectrometry method that avoids nitration artifacts
Autore:
Frost, MT; Halliwell, B; Moore, KP;
Indirizzi:
Univ London, Guys Kings & Thomas Sch Biomed Sci, Int Antioxidant Res Ctr, London SE1 9RT, England Univ London London England SE1 9RT dant Res Ctr, London SE1 9RT, England Natl Univ Singapore, Dept Biochem, Singapore 119260, Singapore Natl Univ Singapore Singapore Singapore 119260 ngapore 119260, Singapore Univ Coll London, Royal Free Hosp & Univ Coll Med Sch, Dept Med, London NW3 2QG, England Univ Coll London London England NW3 2QG ept Med, London NW3 2QG, England
Titolo Testata:
BIOCHEMICAL JOURNAL
, volume: 345, anno: 2000,
parte:, 3
pagine: 453 - 458
SICI:
0264-6021(20000201)345:<453:AOFAPN>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
NITRIC-OXIDE; TYROSINE NITRATION; 3-NITROTYROSINE; QUANTIFICATION; PEROXYNITRITE; INFLAMMATION; SCLEROSIS; HPLC;
Keywords:
alkaline hydrolysis; nitric oxide; oxidative stress; reactive-nitrogen species; tyrosine;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
24
Recensione:
Indirizzi per estratti:
Indirizzo: Moore, KP Univ London, Guys Kings & Thomas Sch Biomed Sci, Int AntioxidantRes Ctr, St Thomas St, London SE1 9RT, England Univ London St Thomas St London England SE1 9RT E1 9RT, England
Citazione:
M.T. Frost et al., "Analysis of free and protein-bound nitrotyrosine in human plasma by a gas chromatography/mass spectrometry method that avoids nitration artifacts", BIOCHEM J, 345, 2000, pp. 453-458

Abstract

Measurement of nitrotyrosine in biological fluids and tissues is increasingly being used to monitor the production of reactive nitrogen species in vivo. The detection of nitrotyrosine in vivo has been reported with the use of a variety of methods including immunoassay, HPLC and GLC/MS. The validityof HPLC and immunoassays have been questioned with regard to their selectivity and sensitivity limits. In principle, the measurement of nitrotyrosineby GLC/MS permits a highly specific, highly sensitive and fully quantitative assay. The nitration of tyrosine under acidic conditions in the presenceof nitrite is well documented. Derivatization for the full quantification of nitrotyrosine by using GLC/MS can lead to the artifactual nitration of tyrosine if performed under acidic conditions in the presence of nitrite. Wedescribe a novel alkaline method for the hydrolysis and derivatization of nitrotyrosine and tyrosine, and demonstrate its applicability to the measurement of plasma concentrations of both free and protein-bound nitrotyrosineand tyrosine. A detection limit of 1 pg for nitrotyrosine and 100 pg for tyrosine has been achieved, Our method allows, for the first time, the analysis of free and protein-bound nitrotyrosine and tyrosine in biological samples. The plasma concentrations (means +/- S.E.M.) of free tyrosine and nitrotyrosine in eight normal subjects were 12 +/- 0.6 mu g/ml and 14 +/- 0.7 ng/ml respectively. Plasma proteins contained tyrosine and nitrotyrosine at 60.7 +/- 1.7 mu g/mg and 2.7 +/- 0.4 ng/mg respectively.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 22:11:49