Reactions of 1-methyl-2-phenylindole with malondialdehyde and 4-hydroxyalkenals.: Analytical applications to a colorimetric assay of lipid peroxidation
Reactions of 1-methyl-2-phenylindole with malondialdehyde and 4-hydroxyalkenals.: Analytical applications to a colorimetric assay of lipid peroxidation
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DOI：
10.1021/tx9701790
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发表时间：
1998-10-01
影响因子：
4.1
通讯作者：
Chaudière, J
中科院分区：
文献类型：
--
作者：
Gérard-Monnier, D;Erdelmeier, I;Chaudière, J
在酸性和温和条件下，1-甲基-2-苯基吲哚与丙二醛（MDA）和4-羟基醛反应生成稳定的发色团，最大吸光度为586 nn。甲基磺酸的使用使丙二醛和4-羟基壬烯醛的发色团产量达到最佳。相比之下，盐酸的使用导致丙二醛产生的发色团的最佳产量和可忽略不计的4-羟基壬烯醛反应。利用这种显色反应，我们开发了一种新的脂质过氧化比色法。利用甲磺酸基介质，可以在586 nm波长处测量MDA和4-羟基烯醛。然而，与4-羟基烯醛反应的内源性抑制剂的存在是常见的，这意味着后者在一些生物样品中可能被低估。在以盐酸为基础的培养基中进行的测定能够在4-羟基烯醛存在的情况下对MDA进行特异性测量。当希夫碱在盐酸（pH 1.5）中水解时，由于4-羟基醛在盐酸的水解条件下发生不可逆的环化反应，因此两种方法均可用于特异性测量生物样品中总丙二醛的量。将这两种方法应用于脂质过氧化体外模型中单独测定MDA和MDA与4-羟基烯醛的量。该方法还用于阐明在阿霉素处理的啮齿动物中，在希夫碱水解后，体内组织特异性MDA生成的复杂模式。
Under acidic and mild-temperature conditions, 1-methyl-2-phenylindole was found to react with malondialdehyde (MDA) and 4-hydroxyalkenals to yield a stable chromophore with intense maximal absorbance at 586 nn. The use of methanesulfonic acid results in optimal yields of chromophore produced from MDA as well as from 4-hydroxynonenal. By contrast, the use of hydrochloric acid results in an optimal yield of chromophore produced from MDA and a negligible reaction of 4-hydroxynonenal. Taking advantage of such chromogenic reactions, we developed a new colorimetric assay of Lipid peroxidation. Using a methanesulfonic acid-based medium, MDA and 4-hydroxyalkenals can be measured at the 586 nm wavelength. However, the presence of endogenous inhibitors of the reaction with 4-hydroxyalkenals is common, and this means that the latter may be underestimated in some biological samples. The assay performed in a hydrochloric acid-based medium enables the specific measurement of MDA in the presence of 4-hydroxyalkenals. Upon hydrolysis of Schiff bases in hydrochloric acid (pH 1.5), either assay can be used to specifically measure the amount of total MDA in biological samples because 4-hydroxyalkenals undergo an irreversible cyclization reaction under the hydrochloric acid-based conditions of hydrolysis. The two assays were applied to the determination of the amount of MDA alone and of MDA and 4-hydroxyalkenals in an in vitro model of Lipid peroxidation. This methodology was also used to clarify complex patterns of tissue-specific MDA production in vivo, following hydrolysis of Schiff bases, in rodents treated with doxorubicin.