四甲基联苯胺显色光度法测定过氧化氢酶活性

基于Fe²⁺-H₂O₂-四甲基联苯胺(tetramethylbenzidine,TMB)显色体系测定过氧化氢酶(CAT)活性。在Fe ²⁺催化作用下过氧化氢与四甲基联苯胺发生显色反应。加入CAT后,A值减小,显然CAT对显色反应具有阻抑作用。在优化实验条件下,CAT活性在0.00~0.10 U·mL^-1之间与△A呈现良好的线性关系:△A=0.0348+19.19 E(U·mL^-1)(r=0.9977),检出限为0.0016 U·mL^-1。采用酶失活样品液作空白,可有效排除样品共存还原性物质的干扰;方法用于健康人体血清CAT活性检测,为广大临床检验工作者提供参考。

Determination of catalase activity by tetramethylbenzidine spectrophotometric method

In this paper,the activity of catalase was determined based on the Fe²⁺-H 2O₂-tetramethylbenzidine color system.The chromogenic reaction between tetramethylbenzidine and hydrogen peroxide could be occurred under Fe²⁺catalysis.A was decreased with the addition of CAT,herein,CAT had inhibitory effect on color reaction.Under optimal conditions,the CAT activity had a good linear relationship with△A in 0.00~0.10 U·mL^-1,△A=0.0348+19.19 E(U·mL^-1)(r=0.9977),and detection limit of CAT activity was 0.0016 U·mL^-1.Using enzyme inactivation sample solution as blank could effectively eliminate the interference of coexisting reducing substances in the sample.The method was used to detect CAT activity in healthy human serum,which could provide reference for clinical laboratory workers.