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Abstract：
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.
摘要：
一种同时测定天冬氨酸（Asp）的方法，谷氨酸(Glu),甘氨酸(Gly),首次建立了二维液相色谱(2D-LC)结合紫外检测动物血液和脑中牛磺酸(Tau)和γ-氨基丁酸(GABA)的方法。首先,氨基酸神经递质（AANT）用4-氟-7-硝基苯并呋喃（NBD-F）标记在相应的荧光衍生物上，富集在萃取柱上,并自动转移到分析柱上,实现目标组分的在线萃取和完全分离。该方法具有良好的选择性,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。该系统实现了对五种AANT的快速检测和稳定性定量,证明了替代稀释方法的可行性。结果表明,该系统具有较高的承载能力,出色的分辨率,和良好的峰形，不受其他内源性物质的影响。此外，所开发的方法已成功应用于大鼠和猪的血液和全脑中的生物样品的分析。年夜鼠海马和皮质中AANTs的含量高于猪。该方法有望为药理学中AANTs的测定提供适用性。神经科学的药物和临床研究。
