高效液相色谱-电化学检测法测定脑脊液中的多巴胺-β-羟化酶
Determination of dopamine-beta-hydroxylase in cerebrospinal fluid by high-performance liquid chromatography with electrochemical detection.
作者信息
Dillen L, Claeys M, De Potter W P
出版信息
J Pharmacol Methods. 1986 Feb;15(1):51-63. doi: 10.1016/0160-5402(86)90005-7.
An improved method is described for the measurement of dopamine-beta-hydroxylase (D beta H) activity in cerebrospinal fluid, which is based on an incubation with dopamine at a saturated substrate concentration and quantitation of the reaction product noradrenaline, by high-performance liquid chromatography with electrochemical detection using 3,4-dihydroxynorephedrine as internal standard. Sample workup consists in an ion pair extraction to isolate the catecholamines from the rather complex incubation medium, a cation ion exchange to eliminate the bulk amount of dopamine, and alumina adsorption to concentrate the sample prior to high-performance liquid chromatography. The methodology was used to evaluate some of the characteristics of D beta H in cerebrospinal fluid and the stability of the enzyme. The procedure was also employed to determine the change in the D beta H following drug administration. Intravenously administered yohimbine caused an increase in D beta H activity in cerebrospinal fluid of rabbits as expected from its known alpha 2-antagonist properties.
描述了一种改进的方法，用于测量脑脊液中多巴胺-β-羟化酶（DβH）的活性，该方法基于在饱和底物浓度下与多巴胺孵育，并通过使用3,4-二羟基去甲麻黄碱作为内标的高效液相色谱-电化学检测法定量反应产物去甲肾上腺素。样品处理包括离子对萃取，以从相当复杂的孵育介质中分离儿茶酚胺；阳离子交换，以消除大量的多巴胺；以及氧化铝吸附，以在高效液相色谱分析之前浓缩样品。该方法用于评估脑脊液中DβH的一些特性以及该酶的稳定性。该程序还用于确定给药后DβH的变化。静脉注射育亨宾会导致兔脑脊液中DβH活性增加，这与其已知的α2拮抗剂特性相符。
Zotero 插件