Development of a non-derivatized LC-MS/MS method for simultaneous quantification of ten neurotransmitters in five neuroinflammatory mouse models
Bao Sun,
Zhaodi Xia,
Chen Wang,
Mengyao Wu,
Ru Tie,
Yilin Ru,
Yaxi Meng,
Lu Bai,
Ying Zhang,
Pu Jia,
Shixiang Wang,
Bing Qi,
Xiaohui Zheng
Affiliations
Bao Sun
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China; Department of Pharmacy, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, 710038, PR China
Zhaodi Xia
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China
Chen Wang
College of pharmacy, Xi'an Medical University, Xi'an, 710021, PR China
Mengyao Wu
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China
Ru Tie
Department of Pharmacy, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, 710038, PR China
Yilin Ru
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China
Yaxi Meng
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China
Lu Bai
Instrument Analysis Center, Xi'an Jiaotong University, 28 Xianning West Road, Xi'an 710048, China
Ying Zhang
Instrument Analysis Center, Xi'an Jiaotong University, 28 Xianning West Road, Xi'an 710048, China
Pu Jia
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China
Shixiang Wang
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China
Bing Qi
Department of Pharmacy, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, 710038, PR China; Corresponding authors.
Xiaohui Zheng
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, college of Life Science, Northwest University, Xi'an, 710069, PR China; Corresponding authors.
Sensitive and comprehensive measurement of systemic metabolites of Trp, Phe and Glu in biological samples is effective for understanding the pathogenesis of pain, anxiety, depression, PTSD and other neurological diseases. This study developed a LC-MS/MS method for simultaneous monitoring the 10 neurotransmitters in mouse hippocampus, and applied it to investigate their changes in five neuroinflammatory mouse models. The quantitative analysis of 10 neurotransmitters, including Gamma-aminobutyric acid (γ-GABA), l-Glutamate (Glu), l-Glutamine (Gln), l-Tryptophan (Trp), Serotonin (5-HT), 5-Hydroxyindoleacetic acid (5-HIAA), l-Phenylalanine (Phe), l-Tyrosine (L-Tyr), Dopamine (DA), Levodopa (L-DOPA) was achieved within a chromatographic separation time of 4.5 min by the application of ThermoFisher Scientific™ Vanquish™ HPLC coupled to ThermoFisher ScientificTM TSQ QuantisTM mass spectrometer. Optimized chromatographic separation as well as high sensitivity allow the simultaneous analysis and precise quantification of ten neurotransmitters in mouse hippocampus. The limits of detection in sample ranged from 0.3pg (Phe) to 0.1 μg (γ-GABA). Recoveries were between 88.0 to 113.8 %, the mean intra - day and inter - day coefficient of variation of 5.56 % and 4.72 %, respectively. The matrix effect was from 85.1 to 114.9 %. A series of significant changes in five neuroinflammatory mouse models can be detected. γ-GABA, Glu, Gln, DA, Phe, l-Tyr, 5-HIAA were decreased in depression model. Gln and Glu were decreased in PTSD model. 5-HIAA was elevated in anxiety. 5-HIAA and l-Trp were decreased in CFA and SNI model, respectively. In conclusion, a LC-MS/MS method for simultaneous measurement of 10 neurotransmitters in mouse hippocampus was developed and successfully applied to investigation of the changes in five neuroinflammatory mouse models. The method would be expected to provide applicability to the study of the mechanisms of neurological diseases associated with these neurotransmitters.
