自动碳酸酐酶法测定发锌

DETERMINATION OF ZINC IN HAIR BY THE AUTOMATED CARBONIC ANHYDRASE.

目的 :建立准确、特异、试剂稳定的碳酸酐酶自动生化分析法测定发锌。方法 :头发样本经高温灰化处理后 ,释放游离态的锌能使脱辅基的碳酸酐酶复活 ,以乙酸对硝基酚作为酶促反应底物 ,测定碳酸酐酶再激活后的活性评估发锌的含量。结果 :该法线性范围达 6 1.2 μmol/ L,回收率 96 .5 %～ 10 4 .1% ,批内和批间相对标准偏差分别为 2 .3%和 4 .1% ,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 .0 95 ,r=0 .994 ,P>0 .0 5。用本法测定 6 5例健康成人 ,其发锌参考范围 (x± 2 s) :2 .4 6± 1.0 4 μm ol/ g。结论 :该法具有特异、准确、灵敏、试剂稳定等优点 。

Objective:To establish an accurate,specific automated enzymatic method for the determination of hair zinc with stable reagents by means of carbonic anhydrase(CA) reactivation.Methods:After hair handled by high-temperature ashing,Released free zinc was used to reactivate apoCA to catalyze p-nitrophenyl acetate,which as a substrate in this reaction.Evaluation the hair zinc content by determination activity of reactivated carbonic anhydrase.Results:The linearity was up to 61.2μmol/L.The recoveries ranged from 96.5% to 104.1%.The within run and between run RSD was lower than 2.3% and 4.1% respectively.Comparing this method(X) with AAS(Y),we obtained the regression equation Y=0.971X+0.095,r=0.994,P>0.05.The reference value of hair zinc in 56 health was(x±2s):2.46±1.0μmol/g.Conclusion:It is an accurate,sensitive method with stable reagents and suitable for automatic analyzer.