基于液相色谱-串联质谱技术（LC-MS/MS）检测癫痫小鼠毛发中内源性大麻素（2-AG、AEA）的含量

摘要 目的：基于液相色谱-串联质谱技术（LC-MS/MS）对癫痫小鼠毛发中内源性大麻素（2-AG、AEA）进行含量检测。方法：匹鲁卡品腹腔注射方法建立小鼠癫痫模型，采用LC-MS/MS检测毛发中2-AG、AEA的含量，对比分析不同癫痫发作分级小鼠2-AG、AEA含量差异。结果：标准品溶液中AEA、2-AG的保留时间分别为14.3 min，18.1 min，毛发样品中AEA、2-AG的保留时间分别为14.0 min，17.7 min。毛发样品中AEA的检测限、定量限、回收率为0.7 pg/mg、2.1 pg/mg和97.9%，毛发样品中2-AG的检测限、定量限、回收率为3.2 pg/mg、10.9 pg/mg和99.3%，且二者的日内、日间变异系数均低于15%。癫痫发作1级、2级、3级、4级、5级这五个分级癫痫小鼠的AEA含量和2-AG含量依次升高，不同癫痫发作分级小鼠毛发样品中AEA含量和2-AG含量比较（P均<0.05）。结论：LC-MS/MS测定癫痫小鼠毛发中2-AG、AEA的表达量，具有灵敏度更高，样品使用量更少等优点，可大规模样本研究。

Liquid Chromatography and Tandem Mass Spectrometry Determined the Content (2-AG, AEA) Endocenic Cannabinoids in Epileptic Mice

ABSTRACT Objective: The content of endocannabinoids (2-AG, AEA) was examined based on liquid chromatography and tandem mass spectrometry (LC-MS/MS) in epileptic mice. Methods: The mouse epilepsy model was established by pilocarpine peritoneal injection, methanol incubation and solid phase extraction column extraction, qualitatively tested under atmospheric chemical ionizing positive ion mode and multi-reaction monitoring, quantitatively tested 2-AG and AEA content by liquid chromatography and series mass spectrometry, compared the differences in 2-AG and AEA content of different seizure grades of epileptic mice. Results: The retention times of AEA and 2-AG in the standard solution were 14.3 min, 18.1 min, and those of AEA and 2-AG in the hair samples were 14.0 min, 17.7 min, respectively. Hair AEA, 2-AG detection limits were 0.7 pg/mg and 3.2 pg/mg, quantitative limits of 2.1 pg/mg and 10.9 pg/mg, respectively, the recovery rates were 97.9% and 99.3%, respectively, and the day and daytime coefficient of variation was less than 15%. AEA and 2-AG were increased in grade seizures, grade 1, grade 3, grade 4, and grade 2, and AEA and 2-AG were compared in hair samples of different seizure grades (P<0.05). Conclusion: The determination of endocannabinoids (2-AG, AEA) used by LC-MS/MS has the advantages of higher sensitivity and less sample use, and is conducive to large-scale sample research.