催化动力学光度法测定碱性磷酸酶的理论分析与条件优化  

Theoretical studies and conditional optimization of kinetic spectrophotometric determination of alkaline phosphatase

作　　者：冯春梁[1] 李红丹[1] 张振彦[1] 张丽梅[1] 李言[1] 付荣来[1] 

机构地区：[1]辽宁师范大学化学化工学院功能材料化学研究所,辽宁大连116029

出　　处：《辽宁师范大学学报（自然科学版）》2009年第2期195-198,共4页Journal of Liaoning Normal University:Natural Science Edition

基　　金：国家自然科学基金资助项目(60572009);辽宁省科技攻关项目(2005220024);辽宁省科技基金资助项目(99102004);辽宁省教育厅高校重点实验室项目(2008S134)

摘　　要：根据Michaelis-Menten（M-M）方程和朗伯-比尔定律，对检测碱性磷酸酶（ALP）的催化动力学紫外分光光度法进行了理论分析和条件优化．确定了反应的最适酸度条件为pH10．2，温度为25℃．在0．008-0．12U／mL范围内，ALP的浓度与反应速率呈良好的线性关系，线性方程为y=6．1349x＋0．0037（r=0．9997），检测下限为4×10^-4U／mL．利用所研究的方法对某牛奶场的生鲜牛奶中的ALP进行了检测，其相对标准偏差在5％以内，符合微量分析测试的要求，结果令人满意．An enzymatic spectrophotometric method for the determination of alkaline phosphatase （ALP） was proposed based on the Michaelis-Menten equation and Lambert-Beer law. The optimal conditions of reaction were found, and the influence of the pH, reagent contiontration and temperature on the initial rates of process was studied. Under the optimal conditions, the calibration curve of alkaline phosphatase was obtained, which shows that the rate of the reaction is linear to the ALP activity in the range of 0. 008-0.12 U/mL with the detection limit of 0. 000 4 U/mL. This method is simple , sensitive and suitable for the determination of alkaline phosphatase in the milk with satisfactory results.

关 键 词：碱性磷酸酶 分光光度法 催化动力学 牛奶 

分 类 号：TS251.51[轻工技术与工程—农产品加工及贮藏工程]