高效液相色谱法-质谱检测法测定大鼠脊髓突触中神经递质

Determination of neurotransmitter in synapses of rat spinal cord by ultra-high performance liquid chromatography-mass spectrometry

目的采用超高效液相色谱-质谱检测技术建立一种灵敏、稳定的方法,用于测定大鼠脊髓突触体中的神经递质。方法提取大鼠脊髓突触体并衍生处理,经过流动相检测液程序(乙腈-0.1%甲酸-纯水)进行液相色谱分离,正离子源电喷雾离子化,对SD大鼠神经突触体中可能存在的神经递质监测,质谱仪测定。结果本研究制备的大鼠脊髓突触体,呈形态结构完整且具有膜结构的椭圆形颗粒,包含突触小泡、线粒体、细胞浆等。此检测方法能够定性分析大鼠脊髓突触体中神经递质,且具有较强的抗干扰能力,重现性较好(相关系数0.990 0~0.999 9)。结论该方法成功地检测出大鼠脊髓突触体中所含神经递质及相关代谢产物。

Objective A sensitive and stable method was established by ultra-high performance liquid chromatography-mass spectrometry for the determination of neurotransmitters in the synapses of rat spinal cord.Methods To extract rat spinal cord counter and to process derivatives, then conduct the acetonitrile-0.1%formic acid-liquid chromatographic separation of pure water as mobile phase, positive ion source electrospray ionization was used to SD rats synapse for the purpose of neurotransmitter monitoring, with the help of the triple quadrupole mass spectrometry. Results The rat spinal synapse prepared in this study was an oval particle with a complete morphological structure and membrane structure, including synaptic vesicles, mitochondria, cytoplasm, etc. Moreover, this method could qualitatively analyze the neurotransmitters in the synapses of rat spinal cord. This method had a strong anti-interference ability, good reproducibility and correlation coefficient of 0.990 0 ~ 0.999 9. Conclusion This method can successfully detect the neurotransmitters and related metabolites in the synapses of spinal cord in rats.