A method for the simultaneous determination of aspartic acid (Asp), glutamic acid (Glu), glycine (Gly), taurine (Tau) and gamma-aminobutyric acid (GABA) in animal blood and brain by two-dimensional liquid chromatography (2D-LC) combined with ultraviolet detection was established for the first time. First, the amino acid neurotransmitters (AANTs) were labeled on the corresponding fluorescent derivatives with 4-fluoro-7-nitrobenzofurazan (NBD-F), enriched on the extraction column and automatically transferred to the analytical column to achieve on-line extraction and complete separation of the target components. This method exhibited good selectivity, and the correlation coefficients for the analyte calibration curves of were > 0.99. The intra- and inter-day precisions were ≤ 16.03, and the accuracies were in the range of 70.59–116.20%. The system realizes the rapid detection and stability quantification of the five AANTs, which proves that the alternative dilution method is feasible. The results show that the system has high loading capacity, excellent resolution, and good peak shape and is not affected by other endogenous substances. Moreover, the developed method has been successfully applied to the analysis of biological samples in the blood and whole brain of rats and pigs. The content of AANTs in the hippocampus and cortex of rats was higher than that in those of pigs. This method is expected to provide applicability for the determination of AANTs in pharmacological, pharmaceutical and clinical research in nervous science.

二维液相色谱法（2D-首次建立了结合紫外检测的LC）。首先，将氨基酸神经递质（AANT）用4-氟-7-硝基苯并呋喃山酮（NBD-F）标记在相应的荧光衍生物上，在萃取柱上富集并自动转移到分析柱上，以实现在线萃取并完成目标成分的分离。该方法显示出良好的选择性，并且分析物校准曲线的相关系数> 0.99。日内和日间精度≤16.03，精度在70.59–116.20％的范围内。该系统实现了5种AANT的快速检测和稳定性定量，证明了替代稀释方法是可行的。结果表明，该系统具有较高的负载能力，优异的分离度和良好的峰形，并且不受其他内源性物质的影响。而且，所开发的方法已成功地应用于大鼠和猪血液和全脑中生物样品的分析。大鼠海马和皮层中AANT的含量高于猪。该方法有望为神经科学的药理学，药物学和临床研究中AANTs的测定提供适用性。结果表明，该系统具有较高的负载能力，优异的分离度和良好的峰形，并且不受其他内源性物质的影响。而且，所开发的方法已成功地应用于大鼠和猪血液和全脑中生物样品的分析。大鼠海马和皮层中AANT的含量高于猪。该方法有望为神经科学的药理学，药物学和临床研究中AANTs的测定提供适用性。结果表明，该系统具有较高的负载能力，优异的分离度和良好的峰形，并且不受其他内源性物质的影响。而且，所开发的方法已成功地应用于大鼠和猪血液和全脑中生物样品的分析。大鼠海马和皮层中AANT的含量高于猪。该方法有望为神经科学的药理学，药物学和临床研究中AANTs的测定提供适用性。大鼠海马和皮层中AANT的含量高于猪。该方法有望为神经科学的药理学，药物学和临床研究中AANTs的测定提供适用性。大鼠海马和皮层中AANT的含量高于猪。该方法有望为神经科学的药理学，药物学和临床研究中AANTs的测定提供适用性。