辣根过氧化物酶介导的致癌性非氨基偶氮染料苏丹红I的单电子氧化反应。

A one-electron oxidation of carcinogenic nonaminoazo dye Sudan I by horseradish peroxidase.

作者信息

Semanska Marcela, Dracinsky Martin, Martinek Vaclav, Hudecek Jiri, Hodek Petr, Frei Eva, Stiborova Marie

机构信息

Department of Biochemistry, Faculty of Science, Charles University, Prague, Czech Republic.

出版信息

Neuro Endocrinol Lett. 2008 Oct;29(5):712-6.

PMID:18987613

Abstract

OBJECTIVES

The aim of the study was to examine oxidation of carcinogenic Sudan I by peroxidase and characterize the structure of its two major peroxidasemediated metabolites. Another target of the study was to evaluate a mechanism of this oxidation.

METHODS

Thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) with ultraviolet (UV) and visible (VIS) detection was employed for the separation of Sudan I metabolites formed by peroxidase. UV/ VIS-, and mass- spectroscopy as well as nuclear magnetic resonance (NMR) were used to characterize structures of two major Sudan I metabolites.

RESULTS

Peroxidase oxidizes Sudan I by a one electron oxidation to eight products. Two major Sudan I metabolites were isolated by TLC on silica gel and HPLC and structurally characterized. The major product formed during the Sudan I oxidation by peroxidase is Sudan I metabolite M2, which corresponds to a Sudan I dimer molecule. The second major metabolite (M1) is the product of secondary, enzyme independent reactions, being formed from the Sudan I dimer that lost the benzenediazonium moiety.

CONCLUSIONS

The data are the first report on structural characterization of Sudan I metabolites formed by its oxidation with peroxidase.

摘要

目的

本研究旨在检测过氧化物酶对致癌物质苏丹红I的氧化作用，并对其两种主要的过氧化物酶介导代谢物的结构进行表征。本研究的另一个目标是评估这种氧化作用的机制。

方法

采用薄层色谱法（TLC）和配备紫外（UV）和可见光（VIS）检测的高效液相色谱法（HPLC）对过氧化物酶形成的苏丹红I代谢物进行分离。利用紫外/可见光光谱、质谱以及核磁共振（NMR）对两种主要的苏丹红I代谢物的结构进行表征。

结果

过氧化物酶通过单电子氧化作用将苏丹红I氧化为8种产物。通过硅胶薄层色谱法和高效液相色谱法分离出两种主要的苏丹红I代谢物，并对其结构进行了表征。过氧化物酶氧化苏丹红I过程中形成的主要产物是苏丹红I代谢物M2，它相当于一个苏丹红I二聚体分子。第二种主要代谢物（M1）是次级的、非酶促反应的产物，由失去苯重氮部分的苏丹红I二聚体形成。