高效液相色谱法固相酶检测系统测试生物样本中胆碱与乙酰胆碱的研究  

Determination of acetylcholine and choline in biological samples by using high performance liquid chromatography with immobilized enzyme detecting system

作　　者：李炳源[1] 曹蓓蓓[2] 黄彦[2] 邱一华[2] 

机构地区：[1]南通大学公共卫生学院,南通226001 [2]南通大学医学院生理学教研室,南通226001

出　　处：《南通大学学报（医学版）》2006年第6期429-431,共3页Journal of Nantong University(Medical sciences)

基　　金：江苏省高校自然科学基金(04KJA180110);南通市科技项目(S40057)资助项目

摘　　要：以大孔径玻璃微粒为载体,将生物酶化学键合于玻璃体,制备成Φ5×50mm短柱串联于色谱分离柱后,配合电化学检测器,构建了高效液相色谱固相酶检测平台。采用了AtlantisC18反相色谱柱(一种适用于强极性化合物分析的反相色谱柱)成功分离了胆碱与乙酰胆碱,并特异、灵敏地获取检测信号。研究目标组份在AtlantisC18柱的保留特性,比较了流动相不同组成对酶促反应的影响,优化了色谱条件。色谱条件:AtlantisC18柱,电化检测,流动相:Na2HPO450mmol/L,检测限(S/N=2)在1.1￣3.4pmol以下,绝对回收率均>95.50%,样品日内相对标准差<1.49%。方法精密、准确。To immobilize enzyme to micro beads of large pore glasses by chemical bonding reaction and prepared short （Φ5×50mm） immobilized enzyme column. To develope detecting system by connecting the immobilized enzyme column in the chromatographic separating column behind and with electrochemical detector. We separated acetylcholine （Ach）, choline （Ch） using Atlantis C 18 column successfully and obtained response specificically and sensitively. The characteristics of retention of acetylcholine （Ach）, choline （Ch） in Atilntis and Novle Pak C18 column were observed.Effects of difference constitute in mobile phase on enzyme reaction was compared and separating condition was optimized. Chromatography condition： we used Atlantis C18 column and electrochemical detecdor, the mobile phase was Na2HPO4 50 mmol/L ,The detection limiits of the method were 1.1-3.4pmol （S/N=2）. The recoveries were above 95.50% and the relative standard deviation （RSDs）of inter - day were all within 1.49%. The method has advantages of high sensitivity and accuracy.

关 键 词：高效液相色谱 固相酶 电化学检测 胆碱 乙酰胆碱 

分 类 号：R11[医药卫生—公共卫生与预防医学]