Halliwell B, Gutteridge J M, Aruoma O I
Department of Biochemistry, University of London King's College, United Kingdom.
Anal Biochem. 1987 Aug 15;165(1):215-9. doi: 10.1016/0003-2697(87)90222-3.
Hydroxyl radicals, generated by reaction of an iron-EDTA complex with H2O2 in the presence of ascorbic acid, attack deoxyribose to form products that, upon heating with thiobarbituric acid at low pH, yield a pink chromogen. Added hydroxyl radical "scavengers" compete with deoxyribose for the hydroxyl radicals produced and diminish chromogen formation. A rate constant for reaction of the scavenger with hydroxyl radical can be deduced from the inhibition of color formation. For a wide range of compounds, rate constants obtained in this way are similar to those determined by pulse radiolysis. It is suggested that the deoxyribose assay is a simple and cheap alternative to pulse radiolysis for determination of rate constants for reaction of most biological molecules with hydroxyl radicals. Rate constants for reactions of ATP, ADP, and Good's buffers with hydroxyl radicals have been determined by this method.
在抗坏血酸存在的情况下，铁-乙二胺四乙酸络合物与过氧化氢反应生成的羟基自由基会攻击脱氧核糖，形成在低pH值下与硫代巴比妥酸加热时会产生粉红色色原的产物。添加的羟基自由基“清除剂”会与脱氧核糖竞争产生的羟基自由基，并减少色原的形成。可以从颜色形成的抑制作用中推导出清除剂与羟基自由基反应的速率常数。对于多种化合物，通过这种方法获得的速率常数与通过脉冲辐解测定的速率常数相似。有人认为，脱氧核糖测定法是一种简单且廉价的替代方法，可用于测定大多数生物分子与羟基自由基反应的速率常数，以替代脉冲辐解。已通过该方法测定了ATP、ADP和Good缓冲液与羟基自由基反应的速率常数。
