氯化血红素模拟过氧化物酶高效液相色谱柱后衍生法测定过氧化氢和水溶性有机过氧化物

研究了氯化血红素（Ｈｅｍｉｎ）模拟辣根过氧化物酶（ＨＲＰ），对羟基苯乙酸（ＰＨＰＡＡ）作 底物，高效液相色谱（ＨＰＬＣ）柱后衍生荧光法测定Ｈ２Ｏ２和水溶性有机过氧化物的方法。采用 Ｈｅｍｉｎ作催化剂柱后衍生反应的最佳ｐＨ值约为１１，与荧光检测ｐＨ值一致，使得以往ＨＰＬＣ 柱后衍生方法所需的高压泵从３台减少到２台，而一些不能作为ＨＲＰ底物的羟烷基过氧化 物在ｐＨ值≥１０的溶液中迅速水解为Ｈ２Ｏ２从而得到测定。优化了测定Ｈ２Ｏ２和甲基过氧化 氢（ＣＨ３ ＯＯＨ， ＭＨＰ）的条件。最佳条件下 Ｈｅｍｉｎ方法测定 Ｈ２Ｏ２的检测限为 ９．０ × １０－９ ｍｏｌ／Ｌ， 测定 ＭＨＰ的检测限为 ２．０ × １０－７ ｍｏｌ／Ｌ。

Determination of Hydrogen Peroxide and Hydrophilic Organic Peroxides by High -Performance Liquid Chromatography With Hemin as a Peroxidase Substitute

Hemin was used to substitute for horseradish peroxidase(HRP) as catalyst with p-hydroxyphe- nyl acetic acid as a substrate for the detection of hydroperoxides in a high performance liquid chroma- togrpahic (HPLC) system. In hemin-catalyzed system, reaction development and fluorescence measurement can both be conducted optimally in the same ammoniacal buffer, thereby allowing elimination of one of the three pumps perviously required. In addition, some of 1-hydroxyalkyl hydroperoxides which were not suitable for the substrates of HRP can be decomposed rapidly to H2O2 at this pH for the detection. The HPLC system is optimized for the determination of H2O2 and methylhydroperoxide (MHP). The detection Iimit is 9.0 x 10-9 mol/L for H2O2 and 2.0 x 10-7 mol/L for MHP.