用终止剂改进超氧化物歧化酶邻苯三酚测活法

The Improved Pyrogallol Method by Using Terminating Agent for Superoxide Dismutate Measurement

以二硫苏糖醇（ＤＴＴ）或Ｌ－抗坏血酸（ＶｉｔＣ）为终止剂改进的经典邻苯三酚法测定超氧化物歧化酶（ＳＯＤ）活性。反应体系：５０ｍｍｏｌ／Ｌ邻苯三酚，ｐＨ８．２０，总体积９ｍｌ，２５℃，加一滴ＤＴＴ（１００ｍｍｏｌ／Ｌ）或ＶｉｔＣ（５％），约５０μｌ，终止自氧化反应。终止后，反应体系的４２０ｎｍ光吸收在ｌｈ内保持恒定。终止剂法的邻苯三酚自氧化率及酶活测定灵敏度与经典法相近，一个酶活单位相当于纯ＳＯＤ１００μｇ／Ｌ。

The pyrogallol autoxidation methodused for superoxide dismutase measurementhas been improved by using DTT or Vit Cas a terminating agent.The reaction systemconsists of 50mmol/L Tris-HCl,1mmol/L di-ethylene-triaminepentaacedic acid,0.2mmol/Lpyrogallol, pH 8.20 at 25 ℃ in a total volumeof 9ml, one drop(about 50μl)of DTT(100mmol/L)or ascorbic acid( 5%)beingadded to stop the autoxidation reaction,Theabsorbence at 420nm of the reaction systemafter terminating the reaction keeps constantfor an hour.