Open AccessAnalysis of Native Human Plasma Proteins and Haemoglobin for the Presence of Bityrosine by High‐Performance Liquid Chromatography
Author(s) -
Daneshvar Bahrain,
Frandsen Henrik,
Dragsted Lars O.,
Knudsen Lisbeth E.,
Autrup Herman
Publication year - 1997
Publication title -
pharmacology & toxicology
Language(s) - English
Resource type - Journals
eISSN - 1600-0773
pISSN - 0901-9928
DOI - 10.1111/j.1600-0773.1997.tb00047.x
Subject(s) - chemistry , peroxidase , reactive oxygen species , high performance liquid chromatography , blood proteins , hydrolysis , in vivo , biochemistry , tyrosine , albumin , chromatography , oxidative phosphorylation , size exclusion chromatography , enzyme , biology , microbiology and biotechnology
Generation of reactive oxygen species in vivo results in oxidative‐damage to cellular components, including proteins. Due to the relatively long half‐lives of several blood proteins the cumulative formation of oxidatively damaged proteins might serve as a biomarker for reactive oxygen species formation. The most prominent sources of reactive oxygen species in vivo are site‐specific metal ion‐catalyzed reactions of the Fenton and Haber‐Weiss types and the H 2 O 2 /peroxidase system. In vitro oxidation of L‐tyrosine using a peroxidase or Cu ++ /H 2 O 2 system gives rise to the formation of a highly fluorescent substance, bityrosine. High‐performance liquid chromatography (HPLC) analysis of acid hydrolyzed serum albumin after oxidation with peroxidase/ H 2 O 2 or with Cu ++ /H 2 O 2 showed that bityrosine had been formed whereas oxidation of this protein with Fe(III)/ ascorbate did not result in the formation of bityrosine. Bityrosine could not be detected in human plasma proteins or haemoglobin with the detection limit of one pmol per mg protein.