色谱

色谱 ›› 2011, Vol. 29 ›› Issue (02): 146-151.

• 研究论文 • 上一篇    下一篇

高效液相色谱-荧光检测法同时测定大鼠不同脑区中的8种单胺类神经递质

赵燕燕1,2*, 刘丽艳2, 韩媛媛1, 白洁1, 杜光玲1, 高茜2   

  1. 河北大学医学实验中心,河北大学化学与环境科学学院
  • 收稿日期:2010-09-27 修回日期:2010-11-16 出版日期:2011-02-28 发布日期:2007-03-25
  • 通讯作者: 赵燕燕,博士,教授,硕士生导师,主要从事药理学以及将现代分离技术用于临床、药学、食品、环境等方面的研究工作.
  • 基金资助:

    河北省科技攻关项目(No. 05276101D-88)、河北省教育厅科学研究计划项目(No. 2009315)、河北省卫生厅医学科学研究重点课题计划项目(No. 20090570)和河北省中医药管理局科研计划课题(No. 2008072).

Simultaneous detection of 8 monoamine neurotransmitters in the different sections of mouse brains by high performance liquid chromatography with fluorescence detection

ZHAO Yanyan1,2*, LIU Liyan2, HAN Yuanyuan1, BAI Jie1, DU Guangling1, GAO Qian2   

  1. Experimental Center of Medicine, Hebei University, College of Chemistry and Environmental Science, Hebei University
  • Received:2010-09-27 Revised:2010-11-16 Online:2011-02-28 Published:2007-03-25

PDF

750

被引次数

49

摘要： 采用高效液相色谱-荧光检测法同时测定了大鼠不同脑区中的左旋多巴、去甲肾上腺素、肾上腺素、多巴胺、二羟基苯乙酸、5-羟色胺、5-羟基吲哚乙酸及高香草酸8种单胺类神经递质的含量。样品经组织裂解液(0.60 mol/L高氯酸、0.50 mmol/L Na2EDTA、0.1 g/L L-半胱氨酸的混合水溶液)处理后,冷冻离心得到上清液;上清液中加入高氯酸沉淀剂(1.20 mol/L K2HPO4、2.00 mmol/L Na2EDTA的混合水溶液)处理后,冷冻离心、过滤。在该优化的色谱条件下,8种单胺类神经递质在1.25～5000 μg/L范围内线性关系良好(r＞0.9999),最低检出限在0.20～5.00 μg/L之间,平均回收率在94.83%～99.19%之间,相对标准偏差在0.08%～2.51%之间。该方法具有操作简便、快捷、回收率高、检出限低、分离度好、结果准确可靠等优点,可以用于体内8种单胺类神经递质的同时检测与分析。

关键词: 大鼠, 单胺类神经递质, 高效液相色谱, 脑, 荧光检测

Abstract: A method for the simultaneous detection of L-dihydroxyphenylalanine, norepinephrine, epinephrine, dopamine, 3,4-dihydroxyphenylacetic acid, serotonin hydrochloride, 5-hydroxyindole-3-acetic acid and homovanillic acid in the different sections of mouse brains was established by using high performance liquid chromatography (HPLC) with fluorescence detection and isocratic elution. Before analysis, the sample was deproteinized by 0.60 mol/L perchloric acid, followed by adjusting pH value of the sample with 1.20 mol/L K2HPO4, addition of 0.1 g/L L-cysteine as antioxidant and 0.50 mmol/L Na2EDTA as complexing agent. The separation column was a Shim-pack C18 column (250 mm×4.6 mm, 5 μm) and the mobile phase (pH 3.8) was 13% methanol containing 50 mmol/L citric acid, 50 mmol/L sodium acetate, 0.5 mmol/L 1-heptanesulfonic acid sodium salt, 5 mmol/L triethylamine and 0.5 mmol/L Na2EDTA. The flow rate was 1.0 mL/min. The injection volume was 10 μL. The emission and excitation wavelengths were 330 nm and 280 nm, respectively. Under the optimized separation conditions, the calibration curves showed good linearity within the concentrations of 1.25~ 5000 μg/L (r＞0.9999). The limits of detection were between 0.20~5.00 μg/L, the average recoveries were between 94.83% and 99.19%, and the relative standard deviations (RSDs) were between 0.08% and 2.51%. The advantages of the method include easy and prompt operation, high recovery, low detection limit, good separation effect, high accuracy and precision. The method has practical value for detecting 8 monoamine neurotransmitters in biological samples.

Key words: brain, fluorescence detection, high performance liquid chromatography (HPLC), monoamine neurotransmitter, mouse