A SPECTROPHOTOMETRIC MICROTITER-BASED ASSAY FOR THE DETECTION OF HYDROPEROXY DERIVATIVES OF LINOLEIC-ACID

被引：97

作者：

AUERBACH, BJ ^{[1
]}

KIELY, JS ^{[1
]}

CORNICELLI, JA ^{[1
]}

机构：

[1] WARNER LAMBERT PARKE DAVIS,PARKE DAVIS PHARMACEUT RES DIV,2800 PLYMOUTH RD,ANN ARBOR,MI 48106

来源：

ANALYTICAL BIOCHEMISTRY | 1992年 / 201卷 / 02期

关键词：

D O I：

10.1016/0003-2697(92)90354-A

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An assay for the detection of hydroperoxy derivatives of linoleic acid formed by the action of 15-lipoxygenase is described. The assay developed is based on a method first reported by Ohishi et al. (1985) Biochem. Int. 10, 205-211) with some important modifications. The assay described herein takes advantage of the ability of (9Z,11E)-13-hydroperoxyoctadecadienoic acid (13-HPODE), the product of the action of 15-lipoxygenase on linoleic acid, to oxidize N-benzoyl leucomethylene blue to methylene blue in the presence of hemoglobin. The resultant blue color is stable to light and air and can be quantified spectrophotometrically at 660 nm. The linear range of the assay is 1.6-32 nmol (0.5-10 μg) of 13-HPODE. The ulitity of the assay can be extended to detect other peroxides as well as inhibitors of 15-lipoxygenase. The assay is a rapid, reliable method for the detection of lipid hydroperoxide production. © 1992.

引用

页码：375 / 380

页数：6

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