Target-based metabolomics for the quantitative measurement of 37 pathway metabolites in rat brain and serum using hydrophilic interaction ultra-high-performance liquid chromatography–tandem mass spectrometry

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• 作者：Jiahui Chen ; Waner Hou ; Bo Han ; Guanghui Liu…
• 关键词：Target ; based metabolomics ; Quantitative ; HILIC–MS ; Amino acids ; neurotransmitters ; purines and pyrimidines
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：408
• 期：10
• 页码：2527-2542
• 全文大小：804 KB
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• 作者单位：Jiahui Chen (1)
  Waner Hou (1)
  Bo Han (2)
  Guanghui Liu (1)
  Jin Gong (1)
  Yemeng Li (3)
  Danmin Zhong (3)
  Qiongfeng Liao (1)
  Zhiyong Xie (3) (4)
  
  1. School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou, 510407, China
  2. School of Pharmacy, Shihezi University, Shihezi, 832000, China
  3. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, 510006, China
  4. Cooperative Innovation Center for Molecular Target New Drug Study, University of South China, Hengyang, Hunan, 421001, China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Food Science
  Inorganic Chemistry
  Physical Chemistry
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1618-2650

文摘

Amino acids, neurotransmitters, purines, and pyrimidines are bioactive molecules that play fundamental roles in maintaining various physiological functions. Their metabolism is closely related to the health, growth, development, reproduction, and homeostasis of organisms. Most recently, comprehensive measurements of these metabolites have shown their potential as innovative approaches in disease surveillance or drug intervention. However, simultaneous measurement of these metabolites presents great difficulties. Here, we report a novel quantitative method that uses hydrophilic interaction ultra-high-performance liquid chromatography–tandem mass spectrometry (HILIC-UPLC–MS/MS), which is highly selective, high throughput, and exhibits better chromatographic behavior than existing methods. The developed method enabled the rapid quantification of 37 metabolites, spanning amino acids, neurotransmitters, purines, and pyrimidines pathways, within 6.5 min. The compounds were separated on an ACQUITY UPLC® BEH Amide column. Serum and brain homogenate were extracted by protein precipitation. The intra- and interday precision of all of the analytes was less than 11.34 %, and the accuracy was between −11.74 and 11.51 % for all quality control (QC) levels. The extraction recoveries of serum ranged from 84.58 % to 116.43 % and those of brain samples from 80.80 % to 119.39 %, while the RSD was 14.61 % or less for all recoveries. This method was used to successfully characterize alterations in the rat brain and, in particular, their dynamics in serum. The following study was performed to simultaneously test global changes of these metabolites in a serotonin antagonist p-chlorophenylalanine (PCPA)-induced anxiety and insomnia rat model to understand the effect and mechanism of PCPA. Taken together, these results show that the method is able to simultaneously monitor a large panel of metabolites and that this protocol may represent a metabolomic method to diagnose toxicological and pathophysiological states.