色谱

色谱

• 研究论文 • 上一篇    下一篇

鼠脑组织中6-羟基-1-甲基-1,2,3,4-四氢-β-咔啉及其相关生物胺的高效液相色谱-电化学检测

毛健,苏旸,栾玉静,陈薛钗,邓玉林*   

  1. School of Life Science and Technology, Beijing Institute of Technology, Beijing 100081, China
  • 收稿日期:2008-07-08 修回日期:2008-09-08 出版日期:2009-03-30 发布日期:1982-12-25
  • 通讯作者: 邓玉林
  • 基金资助:
    北京市科技计划项目(20070639034)

Determination of 6-hydroxy-1-methyl-1,2,3,4-tetrahydro-β- carboline, 5-hydroxytryptamine and 5-hydroxyindole acetic acid in the neonatal rat brains using high performance liquid chromatography-electrochemical detection

MAO Jian, SU Yang, LUAN Yujing, CHEN Xuechai, DENG Yulin*   

  1. School of Life Science and Technology, Beijing Institute of Technology, Beijing 100081, China
  • Received:2008-07-08 Revised:2008-09-08 Online:2009-03-30 Published:1982-12-25
  • Contact: DENG Yulin

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530

被引次数

18

摘要： 建立了大鼠脑组织中6-羟基-1-甲基-1,2,3,4-四氢-β-咔啉(6-OH-MTHβC)、5-羟色胺(5-HT)和5-羟吲哚乙酸(5-HIAA)含量的高效液相色谱-库仑阵列电化学检测(HPLC-ECD)方法。采用的色谱柱为DiscoveryHS F5柱(250 mm×4.6 mm,5 μm),流动相为缓冲液（40 mmol/L柠檬酸+20 mmol/L磷酸氢二钠+0.3 mmol/L乙二胺四乙酸二钠,pH 4.0）-甲醇（体积比为78∶22）混合液,流速为1 mL/min。6-OH-MTHβC、5-HT、5-HIAA在1.0～500.0 μg/L范围内线性关系良好(r>0.9992),检出限分别为0.56，0.26，0.53 μg/L,日内和日间精密度(以相对标准偏差表示)均低于6.1%,回收率分别为87.1%～98.2%,87.0%～95.3%,90.1%～97.7%。用该方法检测新生7 d的SD胎鼠脑内6-OH-MTHβC及5-HT、5-HIAA的含量，发现SD胎鼠在急性酒精中毒8 h后6-OH-MTHβC显著上升(P<0.05);而5-HT和5-HIAA的含量有所下降，但无显著性差异。该法简便、稳定、灵敏度高,适用于测定鼠脑组织中6-OH-MTHβC和5-HT,5-HIAA含量的相关研究。

关键词: 5-羟色胺, 5-羟吲哚乙酸, 6-羟基-1-甲基-1,2,3,4-四氢-β-咔啉, 电化学检测, 高效液相色谱, 鼠脑

Abstract: A simple method was developed for the analysis of 6-hydroxy-1-methyl-1,2,3,4-tetrahydro-β-carboline (6-OH-MTHβC), 5-hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) in rat brain by high performance liquid chromatography with electrochemical detection (HPLC-ECD). The separation of the sample was performed on a Discovery HS F5 column (250 mm×4.6 mm, 5 μm) with a mobile phase of the buffer（40 mmol/L citric acid+20 mmol/L Na2HPO4+0.3 mmol/L EDTA2Na, pH 4.0）-methanol （78∶22, v/v） at a flow rate of 1.0 mL/min. The electrochemical detector was Coularray Detector-4. This method showed good linearity (r>0.9992) for the quantification of 6-OH-MTHβC, 5-HT and 5-HIAA in the concentration range of 1.0-500.0 μg/L. The limits of detection were 0.56, 0.26 and 0.53 μg/L, respectively. The recoveries of 6-OH-MTHβC, 5-HT and 5-HIAA spiked in rat brain samples were 87.1%-98.2%, 87.0%-95.3%, 90.1%-97.7%, respectively, and the relative standard deviations of intra-day and inter-day determinations were both less than 6.1%. In comparison with the control, the analysis of alcohol-exposed neonatal rat brain samples revealed a significant difference in the level of 6-OH-MTHβC (P<0.05). The method was proved to be simple, highly sensitive, and could be applied in the analysis of 6-OH-MTHβC, 5-HT and 5-HIAA in the rat brain.

Key words: 5-hydroxyindole acetic acid, 5-hydroxytryptamine, 6-hydroxy-1-methyl-1,2,3,4-tetrahydro-β-carboline, electrochemical detection (ECD), rat brain , high performance liquid chromatography (HPLC)