采用高效液相色谱-荧光检测法测定人血浆和大鼠血清中的多巴胺β-羟化酶。

Assay for dopamine beta-hydroxylase in human plasma and rat serum by high-performance liquid chromatography with fluorimetric detection.

作者信息

Lee M K, Nohta H, Ohkura Y

机构信息

Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.

出版信息

J Chromatogr. 1987 Oct 30;421(2):237-44. doi: 10.1016/0378-4347(87)80404-8.

DOI:10.1016/0378-4347(87)80404-8

PMID:3429580

Abstract

A highly sensitive assay for dopamine beta-hydroxylase activity in human plasma and rat serum by high-performance liquid chromatography with fluorimetric detection is described. Norepinephrine, formed enzymatically from the substrate dopamine, and epinephrine (internal standard), after clean-up with a cation-exchange cartridge (Toyopak IC-SP M), are converted into the corresponding fluorescent compounds by reaction with 1,2-diphenylethylenediamine. The derivatives are separated by reversed-phase chromatography on TSK gel ODS-80TM. The detection limit for norepinephrine formed enzymatically is 5 pmol per assay tube.

摘要

描述了一种通过高效液相色谱荧光检测法测定人血浆和大鼠血清中多巴胺β-羟化酶活性的高灵敏度分析方法。以阳离子交换柱（Toyopak IC-SP M）净化后，由底物多巴胺酶促形成的去甲肾上腺素和肾上腺素（内标），通过与1,2-二苯基乙二胺反应转化为相应的荧光化合物。衍生物在TSK凝胶ODS-80TM上通过反相色谱法分离。酶促形成的去甲肾上腺素的检测限为每个测定管5皮摩尔。