期刊名称：Journal of Clinical Biochemistry and Nutrition
印刷版ISSN：0912-0009
电子版ISSN：1880-5086
出版年度：1996
卷号：20
期号：3
页码：181-193
DOI：10.3164/jcbn.20.181
出版社：The Society for Free Radical Research Japan
摘要：In order to develop a simple and reliable assay method for lipid hydroperoxides in serum or plasma, we sought to determine the suitable conditions for direct application of the previously reported colorimetric method using a methylene blue derivative, 10-(N-methylcarbamoyl)-3, 7-(dimethylamino)-phenothiazine, for the measurement of lipid hydroperoxides in a sample without extraction of them with organic solvents. For such purpose, dissociation of lipid hydroperoxides from proteins by lipoprotein lipase was found to be necessary. Also, we found that holotransferrin, which oxidizes the methylene blue derivative, should be eliminated by its chelation with trimethylenetetraminehexaacetic acid. Pretreatment of the sample with ascorbate oxidase was also necessary to eliminate interference by ascorbic acid in a sample. Thus the recommended method is to mix the sample with lipoprotein lipase, trimethylenetetraminehexaacetic acid, ascorbate oxidase, and the detergent Triton X-100, then to incubate the mixture with the methylene blue derivative dissolved in the detergent in the presence of hemoglobin, and to measure the oxidized product, methylene blue. The amount of lipid hydroperoxides is calculated by use of an external standard, either linoleic acid hydroperoxide or cumene hydroperoxide.
关键词：lipid hydroperoxides;serum;plasma;hemoglobin-methylene blue method
