Analysis of the addition products of α-tocopherol with phosphatidylcholine-peroxyl radicals by high-performance liquid chromatography with chemiluminescent detection

被引：7

作者：

Yamauchi, R

Hara, Y

Murase, H

Kato, K

机构：

[1] Gifu Univ, Fac Agr, Dept Bioproc, Gifu 5011193, Japan

[2] Gifu Univ, Fac Agr, Dept Food Sci, Gifu 5011193, Japan

[3] CCI Corp, Gifu 5013923, Japan

来源：

LIPIDS | 2000年 / 35卷 / 12期

关键词：

D O I：

10.1007/s11745-000-0658-z

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A chemiluminescence-based high-performance liquid chromatographic method was developed for the analysis of the addition products of a-tocopherol with phosphatidylcholine-peroxyl radicals (TOO-PC). The TOO-PC eluted from a reversed-phase column was reacted with a chemiluminescent reagent consisting of a Cypridina luciferin analog and a lipid-soluble iron chelate in acidic methanol at 50 degreesC, and the generated chemiluminescence was monitored. The detection limit for TOO-PC by this method was about 1 pmol. This method was applied to the detection of TOO-PC in the peroxidized membranes prepared from rabbit erythrocyte ghosts. When the erythrocyte ghosts were peroxidized by the addition of a water-soluble free radical initiator, a peak corresponding to TOO-PC was detected on the chromatogram with chemiluminescent detection. The amount of TOO-PC in the erythrocyte membranes increased with the depletion of endogenous a-tocopherol. The results indicate that this method proved useful for the detection of the TOO-PC formed by the peroxyl-radical scavenging reactions of a-tocopherol in biological systems.

引用

页码：1405 / 1410

页数：6

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