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目的从兔血清中提纯对氧磷酶并纯化。方法采用液相色谱的方法,从兔血清中提取对氧磷酶并进行部分纯化。比色法检测待测样品中对氧磷酶的活性,垂直板电泳的方法检测纯化的对氧磷酶的纯度。采用体外解毒的方法,以1.5倍的LD50的敌敌畏(75 mg/kg)同对氧磷酶粗酶(10 U/kg)混合后,给予小鼠腹腔注射,初步验证对氧磷酶的解毒活性。结果纯化后的对氧磷酶比活性为11.54 IU/mg,较纯化前的比活性(0.058 IU/mg)提高了近200倍,产出率约为10%。部分纯化的对氧磷酶对于有机磷类农药有较好的催化水解作用。染毒大鼠的死亡率为0。结论采用液相色谱的方法,可以从兔血清中部分提取并纯化对氧磷酶。
Objective To purify and purify paraoxonase from rabbit serum. Methods Paraoxonase was extracted from rabbit serum and partially purified by liquid chromatography. The colorimetric method was used to determine the activity of paraoxonase in the sample to be tested. The purity of the purified paraoxonase was determined by the vertical plate electrophoresis. In vitro detoxification method, with 1.5 times the LD50 of the dichlorvos (75 mg / kg) with paraoxonase crude enzyme (10 U / kg) mixed, intraperitoneal injection of mice to preliminary verification of paraoxonase detoxification activity . Results The specific activity of purified paraoxonase was 11.54 IU / mg, which was nearly 200 times higher than the specific activity before purification (0.058 IU / mg), and the yield was about 10%. Partially purified paraoxonase for organic phosphorus pesticides have better catalytic hydrolysis. Mortality of the exposed rats was zero. Conclusion The method of liquid chromatography can partially extract and purify paraoxonase from rabbit serum.