Rapid analysis of GABA and glutamate in microdialysis samples using high performance liquid chromatography and tandem mass spectrometry
Rapid analysis of GABA and glutamate in microdialysis samples using high performance liquid chromatography and tandem mass spectrometry
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DOI：
10.1016/j.jneumeth.2009.05.018
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发表时间：
2009-08-30
影响因子：
3
通讯作者：
Ferger, Boris
中科院分区：
文献类型：
--
作者：
Buck, Kerstin;Voehringer, Patrizia;Ferger, Boris
建立了快速、可靠地测定脑微透析液中γ -氨基丁酸（GABA）和谷氨酸的液相色谱-串联质谱（LC-MS/MS）方法。微透析样品分析使用HILIC（亲水相互作用液相色谱）柱，这是能够保留极性氨基酸神经递质。流动相由0.1%甲酸水溶液和乙腈二元梯度洗脱组成。采用三联四极杆质谱仪在电喷雾电离模式下检测GABA、谷氨酸以及各自的内标[D-6]-GABA和[D-5]-谷氨酸，运行时间为3 min。GABA和谷氨酸的线性范围分别为1 ~ 10 μ M和10 nM ~ 10 μ M。GABA的定量限为1 nM，谷氨酸的定量限为10 nM（注射量为10 μ l）。将本方法与传统的高效液相色谱（HPLC）和荧光检测（FD）分析GABA和谷氨酸的方法进行比较。最后，通过监测钾刺激后苍白球中GABA和谷氨酸的细胞外浓度变化，在大鼠体内微透析中验证了LC-MS/MS方法的可行性。(C) 2009 Elsevier B.V.版权所有
A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been established for the rapid and reliable determination of gamma-aminobutyric acid (GABA) and glutamate in brain microdialysates. The microdialysis samples were analysed using a HILIC (hydrophilic interaction liquid chromatography) column, which is able to retain the polar amino acid neurotransmitters. The mobile phase consisted of a binary gradient elution profile comprising 0.1% formic acid in water and acetonitrile. GABA, glutamate as well as the respective internal standards [D-6]-GABA and [D-5]-glutamate were detected by a triple quadrupole mass spectrometer in the positive electrospray ionisation mode within a running time of 3 min. The linearity ranged from 1 nM to 10 mu M for GABA and 10 nM to 10 mu M for glutamate. The limit of quantitation was found to be 1 nM for GABA and 10 nM for glutamate (injection volume 10 mu l). The present LC-MS/MS method was compared to the classical method for analysis of GABA and glutamate using high performance liquid chromatography (HPLC) and fluorescence detection (FD). Eventually, the feasibility of the LC-MS/MS method was demonstrated using in vivo microdialysis in rats by monitoring changes of the extracellular concentrations of GABA and glutamate in the globus pallidus following stimulation with potassium. (C) 2009 Elsevier B.V. All rights reserved.