A fluorometric method for measurement of peroxyl radical scavenging activities of lipophilic antioxidants

被引:120
作者
Naguib, YMA [1 ]
机构
[1] Phytochem Technol, Chelmsford, MA 01824 USA
关键词
lipid peroxidation; antioxidants; liposomes; Trolox; alpha-tocopherol; alpha and beta-carotenes; lutein; probucol;
D O I
10.1006/abio.1998.2931
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A fluorometric method for evaluation of activities of lipophilic antioxidants toward peroxyl radicals in a lipophilic medium (octane:butyronitrile; 9:1, v/v) and dioleoylphosphatidyl choline (DOPC) liposomal suspension in Tris-HCl buffer (pH 7.4 at 40 degrees C) has been developed. This fluorometric method is based on the use of the fatty acid 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic-acid (BODIPY 581/591 C-11) as an indicator, 2,2'-azobis-2,4-dimethyl valeronitrile (AMVN) as a peroxyl radical generator, and 6-hydroxy-2,5,7,8-tetramethyl chroman-2-carboxylic acid (Trolox) as a calibrator. In the absence of antioxidants, a linear correlation was found between the fluorescence decay of BODIPY 581/591 C-11 and the concentration of AMVN. Trolox was used as a standard reference material for validating this newly developed method. A linear correlation was found between the net protection of the fluorescence of BODIPY 581/591 C-11 and the amounts of added Trolox. Employing this method, the relative peroxyl radical scavenging activities of Trolox, alpha-tocopherol, alpha-carotene, beta-carotene, lutein, and probucol in octane: butyronitrile (9:1, v/v) were determined to be 1.0:0.7: 0.5:0.3:0.4:0.3. In the DOPC system, beta-carotene showed an antioxidant activity which is a factor of two greater than that of alpha-tocopherol. This assay is suitable for measurement of activities of antioxidants in an organic lipid environment and a liposomal medium. (C) 1998 Academic Press.
引用
收藏
页码:290 / 298
页数:9
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