生体過酸化脂質の超微量分析
化学発光検出-高速液体クロマトグラフィーの開発と応用
1989 年 38 巻 10 号 p. 800-808
詳細
1989 年 38 巻 10 号 p. 800-808
A chemiluminescence-high performance liquid chromatography (CL-HPLC) was developed [T. Miyazawa et al., Anal. Lett., 20, 915, (1987)] for pmol level micro-determination of lipid hydroperoxides present in biological samples such as human blood plasma, and its lipoprotein fractions, tissue organs of experimental animals, and cultured human fetal diploid cells [T. Miyazawa et al., Anal. Lett., 21, 1033, (1988); J. Biochem., 103, 744, (1988)]. The method involves separation of each lipid class from tissue total lipids with normal phase or reversed phase HPLC and post-column detection of hydroperoxide-dependent chemiluminescence that is generated by luminol oxidation during a reaction of lipid hydroperoxides with cytochrome c-heme. The high specificity for their hydroperoxide group enables an assay for a large range of hydroperoxide concentrations, with a detection limit of 10 pmol of phospholipid hydroperoxides, triglyceride hydroperoxides and tocopherol hydroperoxides, respectively.
