There is a growing interest in biological models to investigate the effect of neurotransmitter dysregulation on the structure and function of the central nervous system (CNS) at different stages of development. Zebrafish, a vertebrate model increasingly used in neurobiology and neurotoxicology, shares the common neurotransmitter systems with mammals, including glutamate, GABA, glycine, dopamine, norepinephrine, epinephrine, serotonin, acetylcholine, and histamine. In this study, we have evaluated the performance of liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the multiresidue determination of neurotransmitters and related metabolites. In a first step, ionization conditions were tested in positive electrospray mode and optimum fragmentation patterns were determined to optimize two selected reaction monitoring (SRM) transitions. Chromatographic conditions were optimized considering the chemical structure and chromatographic behavior of the analyzed compounds. The best performance was obtained with a Synergy Polar-RP column, which allowed the separation of the 38 compounds in 30 min. In addition, the performance of LC-MS/MS was studied in terms of linearity, sensitivity, intra- and inter-day precision, and overall robustness. The developed analytical method was able to quantify 27 of these neurochemicals in zebrafish chemical models for mild (P1), moderate (P2), and severe (P3) acute organophosphorus poisoning (OPP). The results show a general depression of synaptic-related neurochemicals, including the excitatory and inhibitory amino acids, as well as altered phospholipid metabolism, with specific neurochemical profiles associated to the different grades of severity. These results confirmed that the developed analytical method is a new tool for neurotoxicology research using the zebrafish model.

在生物学模型中，研究神经递质失调对中枢神经系统（CNS）在不同发育阶段的结构和功能的影响越来越引起人们的兴趣。斑马鱼是一种越来越多地用于神经生物学和神经毒理学的脊椎动物模型，与哺乳动物共享常见的神经递质系统，包括谷氨酸，GABA，甘氨酸，多巴胺，去甲肾上腺素，肾上腺素，5-羟色胺，乙酰胆碱和组胺。在这项研究中，我们评估了液相色谱-串联质谱（LC-MS / MS）对神经递质和相关代谢产物的多残留测定的性能。第一步，以正电喷雾模式测试电离条件，并确定最佳裂解模式，以优化两个选定的反应监测（SRM）过渡。考虑到所分析化合物的化学结构和色谱行为，对色谱条件进行了优化。使用Synergy Polar-RP色谱柱可获得最佳性能，该色谱柱可在30分钟内分离出38种化合物。此外，还从线性，灵敏度，日内和日间精度以及整体鲁棒性方面研究了LC-MS / MS的性能。发达的分析方法能够对斑马鱼化学模型中的27种神经化学物质进行定量分析，包括轻度（P1），中度（P2）和重度（P3）急性有机磷中毒（OPP）。结果显示，突触相关神经化学物质（包括兴奋性氨基酸和抑制性氨基酸）普遍下降，磷脂代谢发生变化，特定的神经化学特征与严重程度不同相关。