一种基于过氧草酸酯化学发光的检测方法，用于评估过氧化氢清除活性，该方法采用9,10-二苯基蒽作为荧光团。

A peroxyoxalate chemiluminescence-based assay for the evaluation of hydrogen peroxide scavenging activity employing 9,10-diphenylanthracene as the fluorophore.

作者信息

Arnous Anis, Petrakis Christos, Makris Dimitris P, Kefalas Panagiotis

机构信息

Laboratory of Chemistry of Natural Products, Mediterranean Agronomic Institute of Chania (MAICh), P.O. Box 85, 73100 Chania, Greece.

出版信息

J Pharmacol Toxicol Methods. 2002 Nov-Dec;48(3):171-7. doi: 10.1016/S1056-8719(03)00055-8.

DOI:10.1016/S1056-8719(03)00055-8

PMID:14986866

Abstract

INTRODUCTION

A simple, rapid, sensitive, and enzyme-free analytical method for estimating scavenging of hydrogen peroxide (H2O2) was developed.

METHODS

Peroxyoxalate chemiluminescence (POCL) was measured, using 9,10-diphenylanthracene as fluorophore.

RESULTS

The chemiluminescence signal was found to be linear in response to increasing amounts of H2O2 in ethyl acetate/acetonitrile (9:1) (r2 = .9990), within a range of concentrations varying from 9.0 to 72.0 microM. In contrast, acetonitrile was highly unsuitable because of poor linearity (r2 = .3736) and poor signal stability. The linearity of POCL inhibition, as a measure of H2O2 scavenging, was tested employing well-known, lipid-soluble antioxidants, including beta-carotene, butylated hydroxytoluene (BHT) and alpha-tocopherol, and also the more polar flavonol quercetin, and the water-soluble L-ascorbic acid (AA). Under the experimental conditions, the corresponding values of H2O2 scavenging activity (SA(HP)) for quercetin, beta-carotene, alpha-tocopherol, and L-AA were 19.1 +/- 0.4, 70.9 +/- 20.1, 8.4 +/- 0.4, and 44.8 +/- 5.6 x 10(-3) microM(-1)

DISCUSSION

The data establish the assay as a method for assessing the H2O2 quenching activity of lipid-soluble antioxidants.

摘要

引言

开发了一种简单、快速、灵敏且无酶的分析方法来评估过氧化氢（H₂O₂）的清除能力。

方法

以9,10 - 二苯基蒽为荧光团，测量过氧草酸酯化学发光（POCL）。

结果

在乙酸乙酯/乙腈（9:1）中，随着H₂O₂量的增加，化学发光信号呈线性响应（r² = 0.9990），浓度范围为9.0至72.0微摩尔。相比之下，乙腈由于线性较差（r² = 0.3736）和信号稳定性差而非常不合适。采用众所周知的脂溶性抗氧化剂，包括β - 胡萝卜素、丁基羟基甲苯（BHT）和α - 生育酚，以及极性更强的黄酮醇槲皮素和水溶性L - 抗坏血酸（AA），测试了作为H₂O₂清除能力衡量指标的POCL抑制的线性。在实验条件下，槲皮素、β - 胡萝卜素、α - 生育酚和L - AA的H₂O₂清除活性（SA(HP)）相应值分别为19.1±0.4、70.9±20.1、8.4±0.4和44.8±5.6×10⁻³微摩尔⁻¹。