Potter P E, Meek J L, Neff N H
J Neurochem. 1983 Jul;41(1):188-94. doi: 10.1111/j.1471-4159.1983.tb13668.x.
A simple, rapid method is presented for the determination of acetylcholine (ACh) and choline (Ch) in neuronal tissue using HPLC with electrochemical detection. The method is based on the separation of ACh and Ch by reverse-phase HPLC and mixing the effluent as it emerges from the column with acetylcholinesterase and Ch oxidase, which converts endogenous Ch and Ch produced by the hydrolysis of ACh to betaine and hydrogen peroxide. Production of hydrogen peroxide is continuously monitored electrochemically. The sensitivity of the procedure is 1 pmol for Ch and 2 pmol for ACh. Specificity of the method is based on HPLC, two specific enzymatic reactions, and the detection of hydrogen peroxide.
本文介绍了一种简单、快速的方法，用于使用高效液相色谱-电化学检测法测定神经元组织中的乙酰胆碱（ACh）和胆碱（Ch）。该方法基于反相高效液相色谱对ACh和Ch的分离，并将柱流出物与乙酰胆碱酯酶和Ch氧化酶混合，后者将内源性Ch以及ACh水解产生的Ch转化为甜菜碱和过氧化氢。通过电化学方法持续监测过氧化氢的产生。该方法对Ch的灵敏度为1皮摩尔，对ACh的灵敏度为2皮摩尔。该方法的特异性基于高效液相色谱、两个特异性酶促反应以及过氧化氢的检测。
