芳环羟基化HPLC分离荧光法检测Cu(Ⅱ)-H_2O_2体系中产生的·OH  

Fluorimetry Assay of Hydroxyl Radical by Cu(Ⅱ)-H_2O_2-Aromatic Hydroxylation System

作　　者：方茹[1] 于怀东[1] 蒋海燕[1] 杨少宗[2] 邹国林[1] 

机构地区：[1]武汉大学生命科学学院,湖北武汉430072 [2]阜阳师范学院,安徽阜阳236041

出　　处：《武汉大学学报（理学版）》2004年第4期525-528,共4页Journal of Wuhan University:Natural Science Edition

基　　金：安徽省高等学校青年教师科研资助计划项目(2004Jql77)

摘　　要：采用L 苯丙氨酸为探针,使用液相色谱分离荧光检测(FLD)和荧光分光光度分析(FS)两种方法平行检测Cu(Ⅱ) H2O2体系中产生的·OH.试验采用的激发波长277nm,发射波长306nm.体系在反应前后的荧光变化,可反映·OH产生量.对FLD与FS所得数据进行了比较分析,结果显示两种方法具有较高一致性.FS使用混合体系检测,易对荧光的产生造成干扰,而FLD法没有干扰.In this study, RP-HPLC with fluorescence detection and Fluorescence spectrum (excitation 277 nm and emission 306 nm) was used to detect hydroxyl radical generated by Cu(Ⅱ)-H_2O_2 (Fenton like reaction) system, using a fluorescence probe, L-phenylalanine, which could be converted to highly fluorescent L-tyrosine in the presence of ·OH. Before and after reaction, the variety of fluorescence intensity reflected the generation of ·OH. The data of two methods were compared, by which, we can conclude that the result of two methods have quite coherence. The fluorescence spectrum is a more convenient method. But the fluorescence intensity detected by fluorescence spectrum reflected all fluorescent components in the system. Other ·OH generation system, such as Fe(Ⅱ)-H_2O_2 or phenanthroline- Cu(Ⅱ) coordinate-H_2O_2,influenced the accuracy of the result,the method of detecting ·OH by FS is confined.

关 键 词：羟自由基 芳环羟基化 荧光法 苯丙氨酸 

分 类 号：Q556[生物学—生物化学]