人血清羧酸酯酶活性测定方法的建立-中国科技论文在线.PDFVIP

环境与职业医学2004年4月第21卷第2期 J Environ Occup Med，Apr 2004 Vo1．21 No．2 · l27 · 文章编号：1006．3617(2004102．0127-04 中图分类号：Q55 文献标识码：A 【论著】 人血清羧酸酯酶活性测定方法的建立 郑捷，孙运光，郑光，周志俊 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(sI)s)终止酶促反应，用分光光度 法测定酶促反应的产物 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—1000 U范围内与吸光度值呈线性关系(r=0．999)，方法的 RSD在1．09％ 6．14％之间。加入不同酶量(25％、50％、75％的血清)，实测值与理论值误差百分比分别为5．17％、 5．31％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。 关键词： 羧酸酯酶；活性；检测 study眦 the Measurement of Serum Carlmxylesterase Activity ZHENG J 1 SUN Yun-guang，zHENG Guang，朋O z n(School ofPublic Health，Fudan University，Shanghai 200032，China) Al~raet [Objective]To establish a method to ure the activity of$ertllffl carboxylestemse using a simple spectropliotometer．[Methods] nle amoImt of a-naphthol liberated upon hydrolysis of ct-naphthyl acetate at 600珈m was measured to present the carboxylestemse activity．The reactive conditions were optimized．The final assay consisted of5td senJIIl，added w油 2．5 TrIl mixture of0．03 mol／L a-naphthyl acetate in PBS，incubated at 37℃for 30 minutes and inhibited by 50 5％SDs．[ResuItsj 111e carboxylestemse activity showed linear correlation with the absorbance value betwecn0 and 1000 units(r=0．999)．The relative standard deviation(RSD)ofthis method Was from 1．09％ to 6．14％．By adding 25％，50％，and 75％ of the enzyme in flelllm，the difference between observed and theoretical values Was 5．17％，5．31％ and 1．82％，respectively． I1le RSD among different daily~ ureinent varied from 5．68％to 11．19％．[Conclusion]These results indicate that the method is good eno with time saving for detecting the carboxyleste