Mao Jian, Su Yang, Luan Yujing, Chen Xuechai, Deng Yulin
School of Life Science and Technology, Beijing Institute of Technology, Beijing 100081, China.
Se Pu. 2009 Mar;27(2):216-9.
A simple method was developed for the analysis of 6-hydroxy-1-methyl-1,2,3,4-tetrahydro-beta-carboline (6-OH-MTHbetaC), 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 x 4.6 mm, 5 microm) 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-MTHbetaC, 5-HT and 5-HIAA in the concentration range of 1.0-500.0 microg/L. The limits of detection were 0.56, 0.26 and 0.53 microg/L, respectively. The recoveries of 6-OH-MTHbetaC, 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-MTHbetaC (P < 0.05). The method was proved to be simple, highly sensitive, and could be applied in the analysis of 6-OH-MTHbetaC, 5-HT and 5-HIAA in the rat brain.
建立了一种通过高效液相色谱-电化学检测法（HPLC-ECD）分析大鼠脑中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）的简单方法。样品在Discovery HS F5柱（250 mm×4.6 mm，5μm）上进行分离，流动相为缓冲液（40 mmol/L柠檬酸+20 mmol/L Na2HPO4+0.3 mmol/L乙二胺四乙酸二钠，pH 4.0）-甲醇（78:22，v/v），流速为1.0 mL/min。电化学检测器为库仑阵列检测器-4。该方法在1.0-500.0μg/L浓度范围内对6-OH-MTHβC、5-HT和5-HIAA的定量具有良好的线性（r>0.9992）。检测限分别为0.56、0.26和0.53μg/L。大鼠脑样品中添加的6-OH-MTHβC、5-HT和5-HIAA的回收率分别为87.1%-98.
