【摘要】 目的　合成酸性磷酸酶 (ACP)的底物 2 ,6 二氯 4 乙酰基苯基磷酸 (DCAP P) ,并建立以DCAP P为底物的一种新的ACP测定方法。方法　在BTS 310型半自动分析仪和TBA 4 0FR型全自动生化分析仪上研究连续监测法 ,建立试验参数。结果　本法测定DCAP吸收峰为 32 6nm ;ACP的Km=0 139mmol/L ;340nm时DCAP的摩尔消光系数为 14 4 4 0L·mol 1·cm 1,缓冲液浓度 0 1mol/L ,最适pH5 .4 ,底物浓度 2 0mmol/L ,延滞期 6 0s,线性反应期 15min ;线性范围为 0～ 1188U/L ;血红蛋白 <115 6 2mg/L及胆红素对测定结果无影响 ;本法总ACP与PAP(前列腺酸性磷酸酶 )的参考值范围分别为 12 5 8～ 19 4 2U/L和 2 5 2～9 0 4U/L。结论　此方法重复性好 ,线性反应期长 ,线性范围广 ,干扰因素少 ,适用于自动化分析仪。更多还原

【Abstract】 Objective 2,6-dichloro-4-acetylphenylphosphoric acid (DCAP-P) was synthesized as the substrate of acid phosphatase (ACP) for determination of activity and a new method was established.Methods By using semiautomatic and automatic analyzer, a continuous-monitoring procedure was investigated and the experimental parameters were set up.Results Based on the results of ACP in 30 health adults by the determination,the wavelength of DCAP’s absorption was 326nm,and the Km of ACP was 0.139mmol/L,the molecular extinction coefficient of DCAP at 340nm wavelength was 14 440L/(mol×cm).The conditions of reaction were as follows:the concentration of citrate buffer was 0.1 mol/L, the optimum pH of enzyme was 5.4,the concentration of DCAP-P was 2.0mmol/L,the delay time was 60s,the time of linear reaction was 15 minutes and the linear range was 0-1188u/L.The concentration of total ACP and PAP (prostatic acid phosphatase) in serum were 12.58 u/L-19.42 u/L and 2.52 u/L-9.04 u/L respectively.Conclusion The method is convenient,reproducible,time-saving for practice and reliable to be unaffected by bilirubin in serum.It is especially suited to the automatic analyzers.更多还原