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测定葡萄糖氧化酶(GOD,E.C.1.1.3.4)最广泛使用的方法,是利用葡萄糖氧化酶—辣根过氧化物酶—邻联茴香胺偶联反应,用分光光度法测定;但邻联茴香胺是致癌物,有必要选择毒性较低、灵敏度较高的新染料代替之。国外曾有报道,邻联甲苯胺、邻甲苯胺及2,6-二氯苯酚等一些染料可作为邻联茴香胺的代用品,但前二种仍有致癌性。国内虽有利用GOD酶法、用4-氨基安替比林偶联酚测定血清中葡萄糖含量(Trinder反应)的报道,并指出Barham及Trinder1972年提出用2.4-二氯苯酚代替酚,灵敏度更高,但直接用以测定GOD活性的方法尚未见报道。本文根据Trinder反应原理,选用2,4-二氟苯酚和苯酚共同做为该偶联反应的氧供体,选择了最佳测定条件,并进行了方法学评价。
The most widely used method for the determination of glucose oxidase (GOD, EC 1.1.3.4) is spectrophotometry using the glucose oxidase-horseradish peroxidase-o-anisidine coupling reaction; but o-anisidine Is a carcinogen, it is necessary to choose less toxic, higher sensitivity of the new dye instead. Foreign countries have reported that o-tolidine, o-toluidine and 2,6-dichlorophenol and some other dyes can be used as o-anisidine substitutes, but the first two are still carcinogenic. Although the domestic use of GOD enzymatic method, using 4-amino-antipyrine coupling phenol in the determination of serum glucose content (Trinder reaction) was reported, and Barham and Trinder in 1997 proposed the use of 2.4-dichlorophenol instead of phenol, higher sensitivity However, no direct method for measuring GOD activity has been reported yet. According to the principle of Trinder reaction, 2,4-difluorophenol and phenol were selected as the oxygen donors for the coupling reaction, the best determination conditions were selected, and the methodological evaluation was conducted.