碳酸酐酶法测定发锌

An new enzymatic method for determination of zinc in hair by the reactivation of carbonic anhydrase

目的:建立灵敏、准确、特异、试剂稳定的碳酸酐酶自动生化分析法测定发锌。方法:头发灰化后溶于0.1mol/LHCl,测定前先用0.01mol/L NaOH稀释并中和酸,测定时待分析液中锌离子复活脱辅基的碳酸酐酶,以醋酸对硝基酚作为酶促反应底物进行测定。结果:线性范围达61.2μmol/L,回收率95.2%～104.9%,批内和批间变异系数分别小于2.1与2.8%,与原子吸收分光光度法(Y)比较具有良好的相关性,Y=0.971X+0.095,r=0.994。发样中Cu2^+、Fe2^+、Mn2^+各25μmol/g、Co2^+1.25μmol/g对本法测定锌均无影响。用本法测定56例健康成人,其发锌参考范围(X±2s):1.16～3.04μmol/。结论:该法具有特异、准确、灵敏、试剂稳定等优点,适合于生化分析仪检测发锌。

objective: To establish a sensitive, accurate, specific automated enzymatic method of determination of hair zinc with stable reagents by means of carbonic anhydrase(CA) reactivation.Methods: After hair handled by high temperature ashing, it was dissolved in 0.1mol/L HCl,then was neutralized by 0.01mol/L NaOH, supernatant fraction was used to reactivate apoCA to catalyze p—nitrophenyl acetate as a substrate.Results:The linearity was up to 61.2μmol/L.The recoveries ranged from 95.2% to 104.9%.The within run and between run CV was lower than 2.1% and 2.8% respectively.Comparing this method (X) with AAS(Y), we obtained the regression equation Y=0.971X+0.095.r=0.994.The assays showed no interference from as much as 25μmol/g hair of Cu2^+、Fe_2^+、Mn_2^+respectively,and 1.25μmol/g hair of Co^(2+).The reference value of hair zinc in 56 health was(X±2s): 1.16～3.04μmol/g.Conclutions:It is a simple, accurate,sensitive method with stable reagents and adapt to automatic analyzer.