采用高效液相色谱-荧光检测法测定单胺氧化酶A和B。

Assay for monoamine oxidases A and B by high-performance liquid chromatography with fluorescence detection.

作者信息

Nohta H, Zaitsu K, Tsuruta Y, Ohkura Y

出版信息

J Chromatogr. 1983 Dec 16;280(2):343-9. doi: 10.1016/s0021-9673(00)91576-0.

Abstract

A sensitive method for the assay of monoamine oxidases A and B is described which employs high-performance liquid chromatography with fluorescence detection. Rat brain mitochondria were used as a preparation of the enzymes. p-Sulfamoylbenzaldehyde and benzaldehyde formed enzymatically from p-sulfamoylbenzylamine (the substrate of monoamine oxidase A) and benzylamine (the substrate of monoamine oxidase B), respectively, are converted simultaneously into fluorescent compounds with 2,2'-dithiobis(1-aminonaphthalene). These compounds are separated by reversed-phase chromatography on mu Bondapak CN. The limits of detection for p-sulfamoylbenzaldehyde and benzaldehyde formed enzymatically are 30 and 10 pmol per assay tube, respectively.

摘要

本文描述了一种用于测定单胺氧化酶A和B的灵敏方法，该方法采用带荧光检测的高效液相色谱法。大鼠脑线粒体用作酶的制备物。分别由对氨磺酰苄胺（单胺氧化酶A的底物）和苄胺（单胺氧化酶B的底物）酶促形成的对氨磺酰苯甲醛和苯甲醛，与2,2'-二硫代双（1-氨基萘）同时转化为荧光化合物。这些化合物通过在μ Bondapak CN上的反相色谱法进行分离。酶促形成的对氨磺酰苯甲醛和苯甲醛的检测限分别为每个测定管30和10皮摩尔。