【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。更多还原

【Abstract】 Superoxide dismutase activity can reflect the ability o f scavenging oxyradical of organism.Determining the activity accurately is ofte n 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 kine tic fluorescent spectrophotometry based on the reaction has been developed to de termine 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 lin ear response range of superoxide dismutase is 0.31-1.58 mg/L,the RSD of seru m sample determination is 1.1%(n=5),and the recoveries of spiked sam ple are between 94.3% and 103.49%.The result of this assay is identical with th at of xanthine-xanthinoxidase assay.This method can be applied to determine th e dismutase activity in the animal tissue sample.更多还原