Simultaneous Measurement of Biogenic Amines and Related Compounds by High Performance Liquid Chromatography (HPLC)

Simultaneous measurement by HPLC was used to determine brain dopamine (DA), noradrenaline (NA), and serotonin (5-HT), and their metabolites, vanillylmandelic acid (VMA), homovanillic acid (HVA), and 5-hydroxyindole acetic acid (5-HIAA). Brain tissues were homogenized with 0.025 N HCl and 0.1 M EDTA. Following extraction with n-butanol, amines were dissolved in acid solution, and the metabolites in alkaline solution; then 5-10 microliters of each was injected into the HPLC simultaneously. The injected samples were detected in the following order at 280 nm: VMA, NA, DA, 5-HIAA, HVA, 5-HT at a flow rate of 2 ml/min within about 20 min. Amines and the metabolites could be separated completely and measured at lower limits of approximately 5-10 ng.