FERROUS ION OXIDATION IN THE PRESENCE OF XYLENOL ORANGE FOR DETECTION OF LIPID HYDROPEROXIDE IN LOW-DENSITY-LIPOPROTEIN
FERROUS ION OXIDATION IN THE PRESENCE OF XYLENOL ORANGE FOR DETECTION OF LIPID HYDROPEROXIDE IN LOW-DENSITY-LIPOPROTEIN
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DOI：
10.1016/0003-2697(92)90122-n
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发表时间：
1992-05-01
影响因子：
2.9
通讯作者：
WOLFF, SP
中科院分区：
文献类型：
--
作者：
JIANG, ZY;HUNT, JV;WOLFF, SP
本文介绍了一种简便、灵敏的直接测定脂蛋白和脂质体中脂质过氧化物的方法。该方法基于在酸性条件下快速过氧化物介导的Fe 2+氧化为Fe 3+的原理。后者在二甲酚橙子的存在下形成Fe 3 +-二甲酚橙子络合物，其可以在560 nm处用荧光光度法测量。用标准过氧化物（如过氧化氢、亚油酸过氧化氢、叔丁基过氧化氢和枯烯过氧化氢）进行校准，得到的平均表观消光系数为4.52 × 104 m − 1 cm − 1，与氢过氧化物氧化亚铁离子的链长约为3一致。内过氧化物在测定中反应性较低或不反应。该方法已在低密度脂蛋白和磷脂酰胆碱脂质体的脂质过氧化研究中得到验证。通过用已知代谢过氧化物的酶进行预处理，我们已经表明该测定法特异性地测量脂质氢过氧化物。测量过氧化反应的其他方法，如共轭二烯，硫代巴比妥酸反应物质和碘量法的评估已与亚铁氧化-二甲酚橙子试验进行了比较。
A simple and sensitive method for the direct measurement of lipid peroxides in lipoprotein and liposomes is described. The method is based on the principle of the rapid peroxide-mediated oxidation of Fe2+to Fe3+under acidic conditions. The latter, in the presence of xylenol orange, forms a Fe3+-xylenol orange complex which can be measured spectrophotometrically at 560 nm. Calibration with standard peroxides, such as hydrogen peroxide, linoleic hydroperoxide, t-butyl hydroperoxide, and cumene hydroperoxide gives a mean apparent extinction coefficient of 4.52 × 104m−1cm−1consistent with a chain length of approximately 3 for ferrous ion oxidation by hydroperoxides. Endoperoxides are less reactive or unreactive in the assay. The assay has been validated in the study of lipid peroxidation of low density lipoprotein and phosphatidyl choline liposomes. By pretreatment with enzymes known to metabolize peroxides, we have shown that the assay measures lipid hydroperoxides specifically. Other methods for measuring peroxidation, such as the assessment of conjugated diene, thiobarbituric acid reactive substances and an iodometric assay have been compared with the ferrous oxidation-xylenol orange assay.