在过量过氧化氢存在的情况下，用辣根过氧化物酶对黄尿酸和烟酸进行荧光衍生化。

Fluorescent derivatization of xanthurenic acid and nicotinic acid with horseradish peroxidase in the presence of excess hydrogen peroxide.

作者信息

Odo Junichi, Sogawa Yuto, Inoguchi Masahiko, Hirai Akihito

机构信息

Department of Biological Chemistry, Faculty of Science, Okayama University of Science, 1-1 Ridai-cho, Kita, Okayama 700-0005, Japan.

出版信息

Anal Sci. 2011;27(1):105-9. doi: 10.2116/analsci.27.105.

DOI:10.2116/analsci.27.105

PMID:21233570

Abstract

The fluorescent derivatization of tryptophan metabolites (xanthurenic acid, nicotinic acid, picolinic acid, and 3-hydroxyanthranilic acid) by the catalytic activity of horseradish peroxidase (HRP) was investigated in the presence of excess H(2)O(2). Non-fluorescent xanthurenic acid (XA) and nicotinic acid (NA) were converted into a fluorescent compound with maximum excitation and emission wavelengths at 325 and 425 nm, and 318 and 380 nm, respectively. This fluorescent derivatization was developed for the fluorometric determination of trace amounts of XA and NA. The calibration curves were linear from 1.0 to 10.0 nmol XA and from 5.0 to 20.0 nmol NA in a 1.0-mL sample solution. The UV spectra of the reaction solutions suggested that compound III as an intermediate of HRP played an essential role in this fluorescent derivatization with HRP.

摘要

在过量过氧化氢存在的情况下，研究了辣根过氧化物酶（HRP）的催化活性对色氨酸代谢产物（黄尿酸、烟酸、吡啶甲酸和3-羟基邻氨基苯甲酸）的荧光衍生化作用。非荧光性的黄尿酸（XA）和烟酸（NA）分别转化为一种荧光化合物，其最大激发波长和发射波长分别为325和425 nm，以及318和380 nm。这种荧光衍生化方法用于荧光测定痕量的XA和NA。在1.0 mL样品溶液中，校准曲线在1.0至10.0 nmol XA和5.0至20.0 nmol NA范围内呈线性。反应溶液的紫外光谱表明，作为HRP中间体的化合物III在HRP的这种荧光衍生化中起着至关重要的作用。