Content Determination of Three Kinds of Monoamine Neurotrasmitters and Their Metabolites in Rat Different Brain Tissue by HPLC with Fluorescence Detection

OBJECTIVE To establish a method for determination of the monoamine neurotrasmitters and their metabolites in brain tissues of the rats by HPLC with fluorescence detection. METHODS Inertsustain C₁₈ column(250 mm×4.6 mm, 5 μm) was used, the mobile phase composed of methanol and water, included 30 mmol citric acid, 40 mmol NaAc, 0.2 mmol EDTA-2Na and 0.4 mmol 1-octanesulfonic acid sodium salt in 1 L water(pH=3.8), gradient elution, the flow rate was 1.0 mL·min^-1, fluorometric detector wave length setting of excitation at 280 nm and emission at 330 nm. RESULTS Norepinephrine, dopamine, serotonin hydrochloride, 2-(4-hydroxy-3-methoxyphenyl)ethylamine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, 5-hydroxyindole-3-acetic acid were completely eluted and well separated in 45 min. Good linear response was obtained for the seven components within the range of 0.9-400.0 μg·L^-1 with correlation coefficients >0.99. The average recovery rate of these components was >80%. CONCLUSION The method is simple, accurate, high separation and good repeatability. It is suitable for measuring the monoamine neurotransmitters and their metabolites in cerebral tissue in common laboratory.