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  • Title: [Simultaneous detection of 8 monoamine neurotransmitters in the different sections of rat brains by high performance liquid chromatography with fluorescence detection].
    Author: Zhao Y, Liu L, Han Y, Bai J, Du G, Gao Q.
    Journal: Se Pu; 2011 Feb; 29(2):146-51. PubMed ID: 21598515.
    Abstract:
    A method for the simultaneous detection of L-dihydroxyphenylalanine, norepinephrine, epinephrine, dopamine, 3,4-dihydroxyphenylacetic acid, serotonin hydrochloride, 5-hydroxyindole-3-acetic acid and homovanillic acid in the different sections of mouse brains was established by using high performance liquid chromatography (HPLC) with fluorescence detection and isocratic elution. Before analysis, the sample was deproteinized by 0.60 mol/L perchloric acid, followed by adjusting pH value of the sample with 1.20 mol/L K2HPO4, addition of 0.1 g/L L-cysteine as antioxidant and 0.50 mmol/L Na2EDTA as complexing agent. The separation column was a Shim-pack C18 column (250 mm x 4.6 mm, 5 microm) and the mobile phase (pH 3.8) was 13% methanol containing 50 mmol/L citric acid, 50 mmol/L sodium acetate, 0.5 mmol/L 1-heptanesulfonic acid sodium salt, 5 mmol/L triethylamine and 0.5 mmol/L Na2EDTA. The flow rate was 1.0 mL/min. The injection volume was 10 microL. The emission and excitation wavelengths were 330 nm and 280 nm, respectively. Under the optimized separation conditions, the calibration curves showed good linearity within the concentrations of 1.25 - 5000 microg/L (r > 0.9999). The limits of detection were between 0.20 - 5.00 microg/L, the average recoveries were between 94.83% and 99.19%, and the relative standard deviations (RSDs) were between 0.08% and 2.51%. The advantages of the method include easy and prompt operation, high recovery, low detection limit, good separation effect, high accuracy and precision. The method has practical value for detecting 8 monoamine neurotransmitters in biological samples.
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