A New Technique for Simultaneous Assay of Biogenic Amines and Their Metabolites in Unpurified Mouse Brain

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Abstract
We developed a chromatographic system which separates cate-cholamines and indolamines as well as a large number of their metabolites (acids, aminoacids and glycols). Waters Radial Pak columns compressed radially to avoid formation of cavities in the packing, were used. These columns must be washed with methanol in order to obtain reproducible results when an ion-pairing mobile phase is employed. The conditioning of bhe columns was not affected by washing with triethylamine or variations in radial pressure. Good resolution of 13 samples was obtained in 28 minutes with a mobile phase composed of 0.1 M KH2PO4, 0.1 mM EDTA and 5 mM heptane sulfonic acid in water and Me OH (85:15), pH = 3.6. Products were identified with the use of a Metrohm 64l electrochemical detector. The technique was used for rapid, simultaneous determination of NA, DA, DOPAC, HVA, 5HT and 5HIAA in deproteinated mouse brain samples.