用于脑透析研究中γ-氨基丁酸（GABA）的高灵敏度高效液相色谱法测定

High sensitivity HPLC assay for GABA in brain dialysis studies.

作者信息

Bourdelais A, Kalivas P W

机构信息

Department of Veterinary Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman 99164-6520.

出版信息

J Neurosci Methods. 1991 Sep;39(2):115-21. doi: 10.1016/0165-0270(91)90077-d.

DOI:10.1016/0165-0270(91)90077-d

PMID:1798342

Abstract

The measurement of GABA in brain dialysis experiments involves precolumn derivatization of the amino acid with o-phthaldehyde (OPA) in the presence of a thiol and subsequent high-pressure liquid chromatography with electrochemical detection. A method is described which employs an internal standard and coulometric preoxidation (+ 0.2 V) to eliminate unconjugated OPA/thiol prior to final oxidation (+ 0.4 V) of the derivatized sample. This allows for more efficient separation, providing a retention time for GABA of 9-10 min, a detection sensitivity of 2 to 5 x 10(-14) mol/sample and chromatographic stability for at least 2 weeks of daily use.

摘要

在脑透析实验中，γ-氨基丁酸（GABA）的测量涉及在硫醇存在的情况下，用邻苯二甲醛（OPA）对氨基酸进行柱前衍生化，随后进行带有电化学检测的高压液相色谱分析。本文描述了一种方法，该方法采用内标和库仑预氧化（+0.2 V），在衍生化样品最终氧化（+0.4 V）之前消除未结合的OPA/硫醇。这使得分离更高效，GABA的保留时间为9 - 10分钟，检测灵敏度为2至5×10⁻¹⁴摩尔/样品，并且在至少两周的日常使用中具有色谱稳定性。