终止剂法测定胆红素氧化酶活性

Improved Method to Measure Bilirubin Oxidase by Using Ascorbic Acid as Terminating Agent

以L-抗坏血酸(Vc)为终止剂改进的经典胆红素氧化酶(BOD)酶活测定方法.反应体系:0.1 mol/L Tris-HCl缓冲液,含50 mmol/L甘露醇,pH 8.1,总体积3 mL,30μL胆红素贮存液,10μL酶液.25℃反应3 min,加入50μL 50 g/L的Vc,终止酶催化反应,440 nm测定光吸收.同时加入空白对照组.终止剂法只需一般的分光光度计,操作方便,数据重复性更高,可以同时测定多个样品.

The normal method for the measurement of bilirubin oxidase was improved by using ascorbic acid as the terminating agent. The reaction system consisted of 0.1 mol/L Tris-HCl（contain 50 mmol/L mannitol）, pH 8.1 ,in a total volume of 3 mL, 30 9L bilirubin solution, 10 9E, BOD. The reaction maintained at 25℃ for 3 min and stopped by adding 50 μL Vc（50 g/L）. At the same time a blank control was made. Using the improved method, the activity of bilirubin oxidase could easily be measured by normal spectrophotometer, espacially multi-samples could also be deal with at the same time.