高效液相电化学法测定大鼠血浆中单胺类神经递质及其代谢产物.docVIP

高效液相-电化学法测定大鼠血浆中单胺类神经递质及其代谢产物 崔翰明，白 鸽，黄世敬，张春光，张秋燕 （中国中医科学院广安门医院，北京 100053） [摘要] 目的：建立高效液相色谱-库仑阵列电化学法测定大鼠血浆中的单胺类神经递质及其代谢产物含量的分析方法。方法：采用Agilent公司的1200型高效液相色谱仪和ESA公司库仑2通道电化学检测器，以3，4二羟基苄胺（DHBA）为内标，测定大鼠血浆中的肾上腺素（E）、去甲肾上腺素（NE）、多巴胺（DA）、高香草酸（HVA）和5-羟色胺（5-HT）的含量。色谱柱：Zorbax Eclipse XDB-C18（4.6mm×150mm, 5μm），加EasyGuard 预柱；流动相：含一水合柠檬酸80mmol?L-1，柠檬酸三钠73.4mmol?L-1，1－辛烷磺酸钠0.12mmol?L-1，乙二胺四乙酸二钠0.1mmol?L-1，18％甲醇的混合溶液，pH4.3；流速：1.0mL?min-1；柱温：25℃；进样量：10μL；测定电极电势：-150mv和400mv。结果：NE、E、DA、HVA和5-HT的最低检测浓度分别为2.0，3.0，5.4，5.2，11.1 ng?mL-1；平均回收率分别为99.72％、85.33％、83.15％、120.43％、103.56％；日内(n=6)和日间(n=3)精密度RSD小于9.12％。结论：本方法操作简便，准确，快速，灵敏度高。可用于大鼠血浆中相关单胺类神经递质的检测和研究。 [关键词] 单胺类神经递质；高效液相色谱；库仑阵列电化学检测器；儿茶酚胺 Measurement of monoamines neurotransmitter and their metabolites in rat plasma by HPLC with Coularray Detaction CUI Han-ming，BAI Ge，HUANG Shi-jing，ZHANG Chun-guang，ZHANG Qiu-yan (Guang’anmen Hospital, China Academy of Chinese Medical Science, Beijing 100053, China) [Abstract] Objective To establish a measurement method on determining the level of monoamines neurotransmitter and their metabolites in rat plasma by HPLC with Coularray Detector-2. Methods The level of NE, E, DA, HVA and 5-HT in rat plasma were determined by Agilent 1200 HPLC with ESA Coularray Detector-2. Mobile phase consisted of 80mmol昄-1 mCitric acid monohydrate, 73.4mmol昄-1 oCitric acid trisodium salt, 0.12mmol昄-1t 1-Octanesulfonic acid sodium salt, 0.1mmol昄-1 oEDTA-Na2 and 18% methanol mixture solution(pH4.3). Flow rate was 1.0 mL?min-1.The Column was Zorbax Eclipse XDB-C18 （4.6mm×150mm, 5μm）and Easyguard guard column at 25℃. The voltage of electrode was -150mv and 【收稿日期】2009-07-03 【通讯作者】崔翰明，Tel：（010-mail：Cui-yaoshi@163.com 【基金项目】国家自然科学基金（编号 400mv. Results The minimum measurement limit of NE, E, DA, HVA and 5-HT were 2.0, 3.0, 5.4, 5.2 and 11.1 ng?mL-1, respectively. The average recoveries were 99.72％, 85.33％, 83.15％, 120.43％ and 103.56％, respectively.Conclusion The method was easy to operate, rap