基于次血红素六肽的过氧化氢和葡萄糖灵敏比色方法  

Hydrogen Peroxide and Glucose Sensitive Colorimetric Method Based on Deuterohemin-Ala His Thr Val Glu Lys

作　　者：石鑫 刘传志[1] 宫平[1] 李伟[1] 侯玥[1] SHI Xin;LIU Chuanzhi;GONG Ping;LI wei;HOU Yue(School of Life Science and Technology,Changchun University of Science and Technology,Changchun 130022,China)

机构地区：[1]长春理工大学生命科学技术学院

出　　处：《应用化学》2019年第7期847-854,共8页Chinese Journal of Applied Chemistry

基　　金：吉林省科技厅科技攻关计划(20170204029SF)资助~~

摘　　要：采用次血红素六肽(DhHP-6)催化H2O2氧化4-氨基安替吡啉-氯代苯酚显色体系,建立了测定H2O2和葡萄糖的方法。研究了pH值、底物浓度和DhHP-6浓度对实验的影响,检测了比色方法的反应线性、稳定性、相关性和回收率。在最佳反应条件下,DhHP-6在不同时间及温度条件下,活性要优于过氧化物酶(POD);DhHP-6催化H2O2的米氏常数(Km)和最大反应速率(vmax)分别为0.171mmo/L和4.22×10^-6mol/s;H2O2响应的线性范围为0.39~25.0mmol/L;高、中、低3水平测定的变异系数(CV)和加标回收率分别在1.29%~2.16%和94.5%~101.1%之间;与葡萄糖商品试剂盒比较相关系数R^2=0.9946;36例血液样品中的葡萄糖浓度在4.26~17.48mmol/L之间。与葡萄糖检测商品试剂盒之间的两组数据经统计差异不显著(P>0.05)。该方法是一种简单、廉价、方便、灵敏的比色测定方法。A simple colorimetric method for determination of H2O2 and glucose was established based on the peroxidase-like activity of deuterohemin-Ala His Thr Val Glu Lys(DhHP-6) to catalyze the oxidation of 4-aminoantipyrine (4-APP)-chlorophenol color reaction system in the presence of H2O2 . The effects of pH, substrate concentration and DhHP-6 concentration were observed, and the reaction linearity, stability, correlation and recovery rate of the colorimetric method were detected. Under optimal reaction conditions, the activity of DhHP-6 is superior to that of peroxidase(POD) under different temperatures and times;the Michaelis constant(Km) and maximum reaction rate(vmax) of DhHP-6 catalyzing H2O2 are 0.171 mmol/L and 4.22×10^-6 mol/s, respectively;the linearity range of H2O2 response is 0.39~25.0 mmol/L, the coefficient of variation(CV) and spiked recovery of the high, medium and low levels are between 1.29%~2.16% and 94.5%~101.1%, respectively;and correlation coefficient with the commercial kit is 0.9946;the concentration of glucose in 36 blood samples are 4.26~17.48 mmol/L. There is no significant difference between the two sets of data for glucose assay kit( P >0.05). Therefore, this is a simple, cheap, convenient and sensitive colorimetric determination method.

关 键 词：过氧化氢 次血红素六肽 模拟酶 比色分析 

分 类 号：O655[理学—分析化学]