摘要
根据酶偶联催化反应机理，通过二次作图法测定了过氧化物酶（Ｓ＿１）和Ｈ＿２Ｏ＿２（Ｓ＿２））的米氏常数（Ｋｍ），分别为０．０６９ｍｍｏｌ／Ｌ和０．１５ｍｍｏｌ／Ｌ。通过理论计算和实验分析确定Ｓ＿１的最适底物浓度为１．５ｍｍｏｌ／Ｌ，Ｓ－２最适底物浓度为２．２５ｍｍｏｌ／Ｌ。实验证明指示酶（Ｇ·Ｒ）用量应＞０．２５Ｕ／ｍｌ反应液，本法采用０．３９Ｕ／ｍｌ反应液并就ｐＨ、温度、Ｈｂ干扰等因素对实验影响进行了系统研究，优选了最佳条件。本法批内变异系数为３．８％，批间变异系数为５．２％。
ccording to mechanism of enzymatic coupled-reaction, Km of GSH (S_1)and H_2O_2(S_2))was mea-sured by double charting method and (km)S_1and(Km)S_2was the 0.069mmol/L,0.15mmol/L re-spectively. Stoicheiometry and experimental analysis verify that the optimal the concentrations of sub-strate Si and S_2 were 1.5mmol/L,and 2.25mmol/L,The result of the experiment have shown that the indicative enzyme(GR)must be provided by>0.23U/ml reaction solutiOn。The concentration of reac-tion solution used in this study was 0.39U/ml.The pH,temperature, Hb,some interference factors af-fecting the experiment were analzyzed and the optimal conditions were selected.CV values of intra-assav was 3.8%,and CV inter-assay 5.2%。
