HPLC-电化学检测法测定大鼠脑内的去甲肾上腺素和5-羟吲哚乙酸  

Determination of Norepinephrine and 5-hydroxyindoleacetic Acid in Rat Brain by HPLC with Electrochemical Detection

作　　者：刘建芳 侯艳宁[1] 吴红海[1] 刘会臣[1] 

机构地区：[1]中国人民解放军白求恩国际和平医院,临床药理室河北石家庄050082

出　　处：《解放军药学学报》2002年第3期143-145,共3页Pharmaceutical Journal of Chinese People's Liberation Army

摘　　要：目的　建立大鼠脑内单胺递质及其代谢产物的测定方法。方法　大鼠脑组织匀浆液用高氯酸沉淀蛋白后 ,上清液直接用于HPLC分析。采用Irregular HC18色谱柱 ,流动相组成为 0 .1mol·L-1NaH2 PO4 甲醇 0 .0 1mol·L-1EDTA - 0 .2mol·L-1辛基磺酸钠 (87∶15∶1∶2 .5 ,pH3.7) ,流速为 0 .8ml/min ,温度为 35℃ ,电化学检测器工作电压为 0 .8V。结果　去甲肾上腺素和 5 羟吲哚乙酸分别在 6 .8～ 6 80ng·ml-1和 3.8～ 380ng·ml-1范围内线性关系良好 ,去甲肾上腺素测定的日内和日间的RSD分别为7.5 %和 8.1% ,5 羟吲哚乙酸测定的日内和日间的RSD分别为 2 .0 %和 7.5 % ,两者回收率分别为 10 3.0 %和 6 1.6 % ,检测限分别可达 2ng·ml-1和 1ng·ml-1。结论　本法用于脑内单胺递质及其代谢物的测定 ,快速、简便。Aim To develop a HPLC method to determine monoamine and its metabolites in rat brain.Methods The supernatants of protein precipitated rat brain tissue homogenates by HClO 4 were directly injected into the chromatographic system.Irregular H C 18 column was used for separation,a mixture of 0.1mol·L -1 NaH 2PO 4,methanol,0.01mol·L -1 EDTA and 0.2mol·L -1 sodium octylsulfonate (87∶15∶1∶2.5) was used as mobile phase,the flow rate was 0.8ml/min,the column temperature was 35℃ and the detector potential was 0.8V.Results Norepinephrine and 5 hydroxyindoleacetic acid have good linearity within the range of 6.8～680ng·ml -1 and 3.8～380ng·ml -1 respectively.Within day and between day precisions of the method were 7.5% and 8.1% for norepinephrine,2.0% and 7.5% for 5 hydroxyindoleacetic acid,respectively.The recoveries of norepinephrine and 5 hydroxyindoleacetic acid were 103.0% and 61.6%,the detection limit was 2ng·ml -1 and 1ng·ml -1 ,respectively.Conclusion The method is fast,simple and accurate in the determination of monoamine and its metabolites in the brain.

关 键 词：HPLC-电化学检测法 高效液相色谱电化学法 去甲肾上腺素 5-羟吲哚乙酸 脑内单胺类递质 测定方法 

分 类 号：R338[医药卫生—人体生理学]