Huang Fei, Li Jia, Shi Hai-Lian, Wang Ting-ting, Muhtar Wahaf, Du Min, Zhang Bei-bei, Wu Hui, Yang Li, Hu Zhi-bi, Wu Xiao-jun
The Ministry of Education (MOE) Key Laboratory for Standardization of Chinese Medicines, The State Administration of TCM (SATCM) Key Laboratory for New Resources and Quality Evaluation of Chinese Medicine, Shanghai Key Laboratory of Complex Prescriptions, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
The Ministry of Education (MOE) Key Laboratory for Standardization of Chinese Medicines, The State Administration of TCM (SATCM) Key Laboratory for New Resources and Quality Evaluation of Chinese Medicine, Shanghai Key Laboratory of Complex Prescriptions, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Department of Pharmacognosy, China Pharmaceutical University, Nanjing 210038, China.
J Neurosci Methods. 2014 May 30;229:8-14. doi: 10.1016/j.jneumeth.2014.04.004. Epub 2014 Apr 13.
There is no method available to simultaneously detect GABA, Glu, Epi, NE, DA, 5-HT and 5-HIAA in mouse hippocampus.
A rapid and sensitive LC-MS/MS method has been developed for simultaneously measuring seven neurotransmitters in mouse hippocampus. The analytes were detected in positive mode with multiple reaction monitoring (MRM) and the procedure was completed in less than 9min.
This method exhibited excellent linearity for all of the analytes with regression coefficients higher than 0.99, and showed good intra- and inter-day precisions (RSD<15%) with good accuracy (80-120%). Moreover, the method was successfully applied for the quantitative determination of neurotransmitters in a mouse depression model induced by successive methylprednisolone injections. The results indicated that this depression model was closely associated with the decreased level of Epi (p=0.002) and elevated ratio of 5-HIAA/5-HT (p=0.01), which has never been reported elsewhere.
COMPARISON WITH EXISTING METHOD(S): Compared with previous methods, current approach is more convenient without any pre-column derivatization of the analytes but enhances detectability with incremental neurotransmitter profile and shortens detection time.
This work represents the first accurate simultaneous determination of seven neurotransmitters in the mouse depression model induced by methylprednisolone. The reliable method will benefit the research of neurological diseases with the altered neurotransmitter profile in brain.
目前尚无方法可同时检测小鼠海马体中的γ-氨基丁酸(GABA)、谷氨酸(Glu)、肾上腺素(Epi)、去甲肾上腺素(NE)、多巴胺(DA)、5-羟色胺(5-HT)和5-羟吲哚乙酸(5-HIAA)。
已开发出一种快速灵敏的液相色谱-串联质谱(LC-MS/MS)方法,用于同时测定小鼠海马体中的七种神经递质。采用多反应监测(MRM)正离子模式检测分析物,整个过程在9分钟内完成。
该方法对所有分析物均表现出优异的线性,回归系数高于0.99,日内和日间精密度良好(相对标准偏差<15%),准确度高(80-120%)。此外,该方法成功应用于连续注射甲基强的松龙诱导的小鼠抑郁模型中神经递质的定量测定。结果表明,该抑郁模型与Epi水平降低(p=0.002)和5-HIAA/5-HT比值升高(p=0.01)密切相关,这在其他地方尚未见报道。
与以前的方法相比,当前方法更方便,无需对分析物进行任何柱前衍生,但通过增加神经递质谱提高了检测能力,并缩短了检测时间。
本研究首次在甲基强的松龙诱导的小鼠抑郁模型中准确同时测定了七种神经递质。该可靠方法将有助于研究脑内神经递质谱改变的神经疾病。