Tufi Sara, Lamoree Marja, de Boer Jacob, Leonards Pim
Institute for Environmental Studies (IVM), VU University Amsterdam, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
Institute for Environmental Studies (IVM), VU University Amsterdam, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
J Chromatogr A. 2015 May 22;1395:79-87. doi: 10.1016/j.chroma.2015.03.056. Epub 2015 Mar 30.
Neurotransmitters are endogenous metabolites that allow the signal transmission across neuronal synapses. Their biological role is crucial for many physiological functions and their levels can be changed by several diseases. Because of their high polarity, hydrophilic interaction liquid chromatography (HILIC) is a promising tool for neurotransmitter analysis. Due to the large number of HILIC stationary phases available, an evaluation of the column performances and retention behaviors has been performed on five different commercial HILIC packing materials (silica, amino, amide and two zwitterionic stationary phases). Several parameters like the linear correlation between retention and the distribution coefficient (logD), the separation factor k and the column resolution Rs have been investigated and the column performances have been visualized with a heat map and hierarchical clustering analysis. An optimized and validated HILIC-MS/MS method based on the ZIC-cHILIC column is proposed for the simultaneous detection and quantification of twenty compounds consisting of neurotransmitters, precursors and metabolites: 3-methoxytyramine (3-MT), 5-hydroxyindoleacetic acid (5-HIAA), 5-hydroxy-L-tripthophan, acetylcholine, choline, L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, epinephrine, γ-aminobutyric acid (GABA), glutamate, glutamine, histamine, histidine, L-tryptophan, L-tyrosine, norepinephrine, normetanephrine, phenylalanine, serotonin and tyramine. The method was applied to neuronal metabolite profiling of the central nervous system of the freshwater snail Lymnaea stagnalis. This method is suitable to explore neuronal metabolism and its alteration in different biological matrices.
神经递质是内源性代谢产物，可实现跨神经元突触的信号传递。它们的生物学作用对许多生理功能至关重要，其水平会因多种疾病而改变。由于神经递质具有高极性，亲水作用液相色谱法（HILIC）是一种很有前景的神经递质分析工具。由于有大量可用的HILIC固定相，因此对五种不同的商用HILIC填充材料（硅胶、氨基、酰胺和两种两性离子固定相）的柱性能和保留行为进行了评估。研究了保留与分配系数（logD）之间的线性相关性、分离因子k和柱分辨率Rs等几个参数，并通过热图和层次聚类分析直观显示了柱性能。提出了一种基于ZIC-cHILIC柱的优化且经过验证的HILIC-MS/MS方法，用于同时检测和定量由神经递质、前体和代谢产物组成的二十种化合物：3-甲氧基酪胺（3-MT）、5-羟基吲哚乙酸（5-HIAA）、5-羟基-L-色氨酸、乙酰胆碱、胆碱、L-3,4-二羟基苯丙氨酸（L-DOPA）、多巴胺、肾上腺素、γ-氨基丁酸（GABA）、谷氨酸、谷氨酰胺、组胺、组氨酸、L-色氨酸、L-酪氨酸、去甲肾上腺素、去甲变肾上腺素、苯丙氨酸、5-羟色胺和酪胺。该方法应用于淡水螺椎实螺中枢神经系统的神经元代谢物谱分析。此方法适用于探索不同生物基质中的神经元代谢及其变化。
