Direct detection of tetrahydrobiopterin (BH4) and dopamine in rat brain using liquid chromatography coupled electrospray tandem mass spectrometry

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• 作者：Hak Rim Kim ; Tae-Hyun Kim ; Sung-Hyun Hong ; Hyung-Gun Kim ; ^{hgkimm@dankook.ac.kr}
• 关键词：Tetrahydrobiopterin ; Dopamine ; Rat brain ; Tandem mass spectrometry
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2012
• 期刊代码：159_0006291x
• 类别：bio
• 出版时间：23 March, 2012
• 卷：419
• 期：4
• 页码：632-637
• 文件大小：661 K

摘要

A simple and rapid liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of tetrahydrobiopterin (BH4) and dopamine in rat brain using epsilon-acetamidocaproic acid (AACA) as an internal standard. Proteins in the samples were precipitated with acetonitrile and then the supernatants were separated by a Sepax Polar-Imidazole (2.1 脳 100 mm, i.d., 3 渭m) column using a mixture of 10 mM ammonium formate in acetonitrile/water (75:25, v/v) as the mobile phase at a flow rate of 300 渭l/min. Quantification was performed on a triple quadrupole mass spectrometer employing electrospray ionization with the operating conditions as multiple reaction monitoring (MRM) and positive ion mode from m/z 242.1 鈫?#xA0;166.0 for BH4, m/z 154.1 鈫?#xA0;90.0 for dopamine and m/z 174.1 鈫?#xA0;114.0 for AACA (IS). The total chromatographic run time was for 5.5 min. The method was validated for the analysis of samples: the limit of detection was 10 ng/g. The calibration curve was linear between 10-2000 ng/g for BH4 (r² = 0.995) and 10-5000 ng/g for dopamine (r² = 0.997) in the rat brain. Thus, good correlated LC-ESI/MS/MS results were obtained and found to be a powerful tool for the quantitative analysis of BH4 and dopamine in the rat brain.