摘要
[目的 ]建立血清羧酸酯酶活性的测定方法。 [方法 ]采用十二烷基磺酸钠 (SDS)终止酶促反应 ,用分光光度法测定酶促反应的产物α 萘酚的生成量 ,以反映羧酸酯酶的活性水平。同时检验其方法的线性范围、相对标准偏差(RSD)、方法的准确性和重现性。 [结果 ]羧酸酯酶活性在 0～ 10 0 0U范围内与吸光度值呈线性关系 (r =0 .999) ,方法的RSD在 1.0 9%～ 6.14 %之间。加入不同酶量 ( 2 5 %、5 0 %、75 %的血清 ) ,实测值与理论值误差百分比分别为 5 .17%、5 3 1%和 1.82 %。同一样本重复测定 ,RSD在 5 .68%～ 11.19%之间。 [结论 ]该检测方法用的血清量少 ,对血清质量要求不是很高 ,酶促反应时间短 ,采用SDS终止反应 ,可以在普通分光光度计上进行测定。实验证明该方法稳定、简便、快速、经济 ,适于实验室研究和现场人群的筛查。
To establish a method to measure the activity of serum carboxylesterase using a simple spectrophotometer. The amount of α-naphthol liberated upon hydrolysis of α-naphthyl acetate at 600 nm was measured to present the carboxylesterase activity. The reactive conditions were optimized. The final assay consisted of 5μl serum, added with 2.5 ml mixture of 0.03 mol/L α-naphthyl acetate in PBS,incubated at 37℃ for 30 minutes and inhibited by 50 μl 5% SDS. The carboxylesterase activity showed linear correlation with the absorbance value between 0 and 1000 units(r=0.999). The relative standard deviation(RSD) of this method was from 1.09% to 6.14%. By adding 25%,50%,and 75% of the enzyme in serum,the difference between observed and theoretical values was 5.17%,5.31% and 1.82%,respectively. The RSD among different daily measurement varied from 5.68% to 11.19%. [Conclusion] These results indicate that the method is good enough with time saving for detecting the carboxylesterase activity and could be widely used in scientific research and population screening since the instrument is simple.
出处
《劳动医学》
北大核心
2004年第2期127-130,共4页
基金
上海市教委基金 (编号 :D/JG0 50 343)
