酶电极L-抗坏血酸速测仪的研制

AN ENZYMED ELECTRODE FOR FAST-DETERMINATION OF L-ASCORBIC ACID

本文研究了黄瓜L-抗坏血酸氧化酶的纯化和固定方法,并研制了利用该酶固定化酶膜和克拉克氧电极构成的L-抗环血酸速测仪。本仪器线性范周在10—1000mg/dl之间,测定20次变异系数为0.79％。响应时间短于20秒。酶膜由基质层,酶层和内层构成,其活性可保持在3用以上。磷酸缓冲液为本仪器所用的唯一试剂。本仪器由一次仪表和二次仪表构成。一次仪表包括酶电极、反应器、磷酸缓冲液系统,该系统还有微量泵和振动搅拌器。二次仪表包括信号的两级放大、信号处理及由计算机控制的功能和数量系统。

The sensors for fast-determination of L-ascorbic acid have been made by immobilizing ascorbate oxidase on cellophane membranes and mounting the enzymic membranes on Clark onygen electrodes and new methods of purifying of L-ascorbate oxidase from the peel of cucumber and immobilizing it. The response of the electrodes is linear between 10 and 1000mg/dl ascorbic acid and the precision is better than 0.79％ by 20 successive assays. The responsing time is less than 20 seconds. The enzymic membrane was made up of three layers, a basic layer, an enzymic layer and an endolayer. And its lifetime is more than 3 weeks. After preparation, buffer of phosphate is the only reagent needed. The sensor involved two parts, the preliminary appearance and the second. The prelimin ary appearance was made of enzyme electrode, actor, a system of phos phate buffer. involving a mirco-pump and a vibrational agitator. And the second, two series magnifying of signal and dealing with the signal as well as function and a system of data display controlling by a computer were involoved.