鲁米诺化学发光的过氧化氢酶活性测定系统

Luminol Chemiluminescence a Novel Catalase Activity Determination System

为了解决传统高锰酸钾法测定过氧化氢酶活性存在的耗时长、准确度低、样本需求量大等问题,开发建立了基于鲁米诺/过氧化氢化学发光的过氧化氢酶类活性测定系统。该系统以鲁米诺被过氧化氢氧化产生化学发光为原理,通过加入过渡金属Co(Ⅱ)和螯合剂EDTA实现持续稳定的高强度化学发光。经实验测定,该系统的鲁米诺化学光强度与过氧化氢的摩尔浓度呈现良好的线性关系;可通过调节检测体系的pH值调整测量灵敏度,进而实现过氧化氢的微量检测。结果表明,该系统对待测蛋白样本量和活性要求较低,有利于微量蛋白的活性检测。同时测定的数据稳定性高,重复性强,准确度高,适用范围广,操作简单,方便快捷,安全性高,值得推广使用。

In order to solve the problem that the traditional potassium permanganate method in the determination of catalase activity has defects of long time consuming,low accuracy and large sample demand,a catalase activity determination system based on luminol luminescence is developed.Based on the measurement of chemiluminescence resulting from the oxidation of luminol with hydrogen peroxide,the transition metal Co(II)and the chelator EDTA is added to optimize the signal with reference to time stability,intensity and reproducibility.The luminescence intensity measured by this new system shows a good linearity with the concentration of hydrogen peroxide,and the activity detection of tiny amounts of catalase can be realized by adjusting the pH value.It is demonstrated that this system has low requirements for the amount and activity of enzyme,which is beneficial to the activity detection of proteins in tiny amounts,and it has the advantages of high data stability,repeatability and accuracy,wide application,simple and fast operation,and high safety,which is worth popularizing.