段文贞, 屈志炜, 张均田. 用反相高效液相色谱—柱后衍生化—电化学检测器测定乙酰胆碱J. 药学学报, 1997, 32(12): 920-923.
引用本文: 段文贞, 屈志炜, 张均田. 用反相高效液相色谱—柱后衍生化—电化学检测器测定乙酰胆碱J. 药学学报, 1997, 32(12): 920-923.
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WZ Duan, ZW Qu , JT Zhang, . DETERMINATION OF ACETYLCHOLINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY, ENZYME-LOADED POSTCOLUMN,AND ELECTROCHEMICAL DETECTORJ. Acta Pharmaceutica Sinica, 1997, 32(12): 920-923.
Citation: WZ Duan, ZW Qu , JT Zhang, . DETERMINATION OF ACETYLCHOLINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY, ENZYME-LOADED POSTCOLUMN,AND ELECTROCHEMICAL DETECTORJ. Acta Pharmaceutica Sinica, 1997, 32(12): 920-923.
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用反相高效液相色谱—柱后衍生化—电化学检测器测定乙酰胆碱

DETERMINATION OF ACETYLCHOLINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY, ENZYME-LOADED POSTCOLUMN,AND ELECTROCHEMICAL DETECTOR

    摘要
  • 摘要: 为了建立灵敏度高、操作方便、适用范围较广的测定乙酰胆碱含量的方法,本文采用高效液相色谱加柱后酶衍生化、电化学检测器方法测定了组织中乙酰胆碱的含量。样本处理采用高氯酸沉淀蛋白后一次性高速离心的上清液进样。样品中乙酰胆碱、胆碱和内标可以基线分离,在流速1.2ml·min-1时,3个峰在10min内全部出完。乙酰胆碱的回收率达85%±2%;样品出峰高度与进样量呈直线关系;重现性好,日内标准偏差为1.2%。结果显示本法可以同时测定组织中胆碱及乙酰胆碱的含量,测定灵敏度可达到1pmol水平。方法简便、快捷、适于在国内实验室推广。

     

    Abstract: A rapid and simple method using a combination of high performance liquid chromatography (HPLC), an immobilized enzyme column, with electrochemical detection (ECD) is described for the assay of acetylcholine (ACh) and choline (Ch) in mouse brain. Perchloric acid extracts of small brain tissue were injected onto the HPLC system with no preclean-up procedure. Detection limits for both compounds were 1 pmol. The present method has been successfully applied to the measurement of ACh in discrete brain areas of the mouse.

     

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