荧光动力学法测定超氧化物歧化酶的活性  

Determination of Superoxide Dismutase Activity by Kinetic Fluorescent Spectrophotometry

作　　者：杨磊[1] 陈冠华[1] 崔建超[2] 郝庆红[1] 李佳[1] 郭云霞[1] 

机构地区：[1]河北农业大学生命科学学院,保定071001 [2]河北农业大学食品科技学院,保定071001

出　　处：《分析化学》2008年第4期489-493,共5页Chinese Journal of Analytical Chemistry

基　　金：河北省自然科学基金(No.202281)资助项目

摘　　要：超氧化物歧化酶的活力可以反映机体清除氧自由基的能力,在抗衰老研究中经常需对其进行准确测定。邻苯三酚自氧化可产生强荧光的醌,超氧化物歧化酶通过清除该反应过程中产生的超氧阴离子自由基使醌减少,据此建立了测定超氧化物歧化酶活力的荧光动力学方法。探讨了缓冲液的浓度、pH值、邻苯三酚浓度和温度对测定的影响。在Tris-HAC缓冲液浓度为0.1mol/L、pH8.2、邻苯三酚浓度为0.385mmol/L、温度为25℃的最佳条件下,得到SOD的线性响应范围为0.31~1.58mg/L;血清样品测定的RSD为1.12%（n=5）;加标回收率在94.3%-103.49%之间。本法与黄嘌呤-黄嘌呤氧化酶法的测定结果一致,可用于动物样品中超氧化物歧化酶活力的测定。Superoxide dismutase activity can reflect the ability of scavenging oxyradical of organism. Determining the activity accurately is often required in anti-aging study. Quinones with hyperfluorescence can be produced from the autoxidation process of pyrogallol. Superoxide dismutase can decrease the quinones by scavenging the superoxide radicals formed in the process. A kinetic fluorescent spectrophotometry based on the reaction has been developed to determine superoxide dismutase activity. The influence of buffer concentration, pH, pyrogallol concentration and temperature on the determination were discussed. The optimum result can be achieved with buffer （tris-HAC） concentration of 0.1 mol/L, and pyrogallol concentration of 0. 385 mmol/L in pH 8.2, at 25℃. The linear response range of superoxide dismutase is 0.31 - 1.58 mg/L, the RSD of serum sample determination is 1.1% （ n = 5） , and the recoveries of spiked sample are between 94.3 % and 103.49%. The result of this assay is identical with that of xanthine-xanthinoxidase assay. This method can be applied to determine the dismutase activity in the animal tissue sample.

关 键 词：邻苯三酚自氧化 超氧化物歧化酶活力 荧光动力学法 

分 类 号：Q55[生物学—生物化学]