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Journal of Analytical Methods in Chemistry
Volume 2014, Article ID 506870, 11 pages
http://dx.doi.org/10.1155/2014/506870
Research Article

Quantification of Neurotransmitters in Mouse Brain Tissue by Using Liquid Chromatography Coupled Electrospray Tandem Mass Spectrometry

1Department of Pharmacology, College of Medicine, Dankook University, Cheonan 330-714, Republic of Korea
2Bioresources Regional Innovation Center, Soon Chun Hyang University, Asan 336-745, Republic of Korea
3Translational Research Center, Institute of Bio-Science Technology, Dankook University, Cheonan 330-714, Republic of Korea

Received 9 June 2014; Revised 18 August 2014; Accepted 21 August 2014; Published 3 September 2014

Academic Editor: Sibel A. Ozkan

Copyright © 2014 Tae-Hyun Kim et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

A simple and rapid liquid chromatography tandem mass spectrometry method has been developed for the determination of BH4, DA, 5-HT, NE, EP, Glu, and GABA in mouse brain using epsilon-acetamidocaproic acid and isotopically labeled neurotransmitters as internal standards. Proteins in the samples were precipitated by adding acetonitrile, and then the supernatants were separated by a Sepax Polar-Imidazole (2.1 mm × 100 mm, i.d., 3 μm) column by adding a mixture of 10 mM ammonium formate in acetonitrile/water (75 : 25, v/v, 300 μl/min) for BH4 and DA. To assay 5-HT, NE, EP, Glu, and GABA; a Luna 3 μ C18 (3.0 mm × 150 mm, i.d., 3 μm) column was used by adding a mixture of 1% formic acid in acetonitrile/water (20 : 80, v/v, 350 μl/min). The total chromatographic run time was 5.5 min. The method was validated for the analysis of samples. The calibration curve was linear between 10 and 2000 ng/g for BH4 , 10 and 5000 ng/g for DA , 20 and 10000 ng/g for 5-HT , NE , and EP , and 0.2 and 200 μg/g for Glu and GABA in the mouse brain tissues. As stated above, LC-MS/MS results were obtained and established to be a useful tool for the quantitative analysis of BH4, DA, 5-HT, NE, EP, Glu, and GABA in the experimental rodent brain.