Fe^(2+)-H_(2)O_(2)-RhB体系荧光猝灭法测定CAT活度  

Determination of Catalase Activity by Fe^(2+)-H_(2)O_(2)-Rhodamine B Fluorescence Quenching System

作　　者：张爱菊 白莹 董娜 薛林科 戴兴德 张小林 ZHANG Aijü;BAI Ying;DONG Na;XUE Linke;DAI Xingde;ZHANG Xiaolin(Gansu Medical College,Pingliang 744000)

机构地区：[1]甘肃医学院,甘肃平凉744000

出　　处：《分析科学学报》2022年第6期793-796,共4页Journal of Analytical Science

基　　金：甘肃省教育厅青年博士基金(2021QB-131),甘肃省高等学校创新基金(2021B-341)。

摘　　要：基于Fe^(2+)-H_(2)O_(2)-RhB荧光猝灭体系实现过氧化氢酶(CAT)活度测定。荧光物质罗丹明B(RhB)于585 nm处有特征荧光峰,H_(2)O_(2)、Fe^(2+)协同作用致其荧光猝灭,CAT是H_(2)O_(2)分解专属酶,CAT对H_(2)O_(2)的前期分解在一定程度上保留了RhB荧光。结果表明,加酶前后荧光强度变化(ΔF)与酶活度之间线性相关,线性范围为5.6×10^(-5)~5.6×10^(-2)U/mL,其线性方程为ΔF=-3.466+1472 E(U/mL),相关系数r=0.9962,检测限为2.6×10^(-5)U/mL。该方法具有良好的重现性和准确性,可应用于人体血清CAT活度的快速检测。In this paper,catalase(CAT)activity was determined based on the Fe^(2+)-H 2O 2-rhodamine B(RhB)fluorescence quenching system.The fluorescent substance of RhB had a characteristic fluorescence peak at 585 nm.The synergistic effect of H_(2)O_(2)and Fe^(2+)resulted in the fluorescence quenching of RhB.CAT is the exclusive enzyme for decomposition of H_(2)O_(2),the predecomposition retained RhB fluorescence to some extent.The results showed that there was a linear equation between fluorescence change(ΔF)and enzyme activity E(U/mL)in the range of 5.6×10^(-5)-5.6×10^(-2)U/mL:ΔF=-3.466+1472 E(U/mL),r=0.9962.The limit of detection was 2.6×10^(-5)U/mL.The method showed good reproducibility and accuracy,indicating that the fluorescence quenching system could be used to detect CAT activity in actual serum.

关 键 词：过氧化氢 过氧化氢酶 罗丹明B 荧光猝灭 

分 类 号：O657.3[理学—分析化学]