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Rapid analysis of glutamate, glutamine and GABA in mice frontal cortex microdialysis samples using HPLC coupled to electrospray tandem mass spectrometry.
- Source :
-
Journal of pharmaceutical and biomedical analysis [J Pharm Biomed Anal] 2018 Apr 15; Vol. 152, pp. 31-38. Date of Electronic Publication: 2018 Jan 31. - Publication Year :
- 2018
-
Abstract
- In vivo measurement of multiple neurotransmitters is highly interesting but remains challenging in the field of neuroscience. GABA and l-glutamic acid are the major inhibitory and excitatory neurotransmitters, respectively, in the central nervous system, and their changes are related to a variety of diseases such as anxiety and major depressive disorder. This study described a simple method allowing the simultaneous LC-MS/MS quantification of l-glutamic acid, glutamine and GABA. Analytes were acquired from samples of the prefrontal cortex by microdialysis technique in freely moving mice. The chromatographic separation was performed by hydrophilic interaction liquid chromatography (HILIC) with a core-shell ammonium-sulfonic acid modified silica column using a gradient elution with mobile phases consisting of a 25 mM pH 3.5 ammonium formate buffer and acetonitrile. The detection of l-glutamic acid, glutamine and GABA, as well as the internal standards [d6]-GABA and [d5]-glutamate was performed on a triple quadrupole mass spectrometer in positive electrospray ionization and multiple reaction monitoring mode. The limit of quantification was 0.63 ng/ml for GABA, 1.25 ng/ml for l-glutamic acid and 3.15 ng/ml for glutamine, and the intra-day and inter-day accuracy and precision have been assessed for the three analytes. Therefore, the physiological relevance of the method was successfully applied for the determination of basal extracellular levels and potassium-evoked release of these neuroactive substances in the prefrontal cortex in adult awake C57BL/6 mice.<br /> (Copyright © 2018 Elsevier B.V. All rights reserved.)
- Subjects :
- Acetonitriles chemistry
Ammonium Compounds chemistry
Animals
Chromatography, High Pressure Liquid methods
Hydrophobic and Hydrophilic Interactions
Male
Mice
Mice, Inbred C57BL
Microdialysis methods
Neurotransmitter Agents chemistry
Spectrometry, Mass, Electrospray Ionization methods
Sulfonic Acids chemistry
Tandem Mass Spectrometry methods
Frontal Lobe chemistry
Glutamic Acid chemistry
Glutamine chemistry
gamma-Aminobutyric Acid chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1873-264X
- Volume :
- 152
- Database :
- MEDLINE
- Journal :
- Journal of pharmaceutical and biomedical analysis
- Publication Type :
- Academic Journal
- Accession number :
- 29414016
- Full Text :
- https://doi.org/10.1016/j.jpba.2018.01.039