title: Quantification of neurotransmitters in mouse brain tissue by using liquid chromatography coupled electrospray tandem mass spectrometry.
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 (r(2) = 0.995) , 10 and 5000 ng/g for DA (r(2) = 0.997) , 20 and 10000 ng/g for 5-HT (r(2) = 0.994) , NE (r(2) = 0.993) , and EP (r(2) = 0.993) , and 0.2 and 200 μg/g for Glu (r(2) = 0.996) and GABA (r(2) = 0.999) 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.
authors.affiliation: Department of Pharmacology, College of Medicine, Dankook University, Cheonan 330-714, Republic of Korea ; Bioresources Regional Innovation Center, Soon Chun Hyang University, Asan 336-745, Republic of Korea.
authors.forename: Tae-Hyun
authors.initials: TH
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, College of Medicine, Dankook University, Cheonan 330-714, Republic of Korea.
authors.forename: Juhee
authors.initials: J
authors.lastname: Choi
authors.affiliation: Department of Pharmacology, College of Medicine, Dankook University, Cheonan 330-714, Republic of Korea ; Translational Research Center, Institute of Bio-Science Technology, Dankook University, Cheonan 330-714, Republic of Korea.
authors.forename: Hyung-Gun
authors.identifier: 0000-0002-7990-3442
authors.initials: HG
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, College of Medicine, Dankook University, Cheonan 330-714, Republic of Korea ; Translational Research Center, Institute of Bio-Science Technology, Dankook University, Cheonan 330-714, Republic of Korea.
authors.forename: Hak Rim
authors.identifier: 0000-0003-3102-2272
authors.initials: HR
authors.lastname: Kim