γ-Aminobutyric acid, catecholamine and indoleamine determinations from the same brain region by high-performance liquid chromatography with electrochemical detection

A new procedure for the measurement of γ-aminobutyric acid, norepinephrine, dopamine, serotonin and 5-hydroxyindoleacetic acid from the same brain region was developed. In general, two separate high-performance liquid chromatographic runs were performed, one for the γ-aminobutyric acid determination and one for the determination of the monoamines. The electrochemical detection of γ-aminobutyric acid was determined by a new procedure that utilized a small aliquot of the brain sample prepared for monoamine measurement. This assay was linear and parallel between 6 and 200 ng per 20-μl injection with 5-aminovaleric acid utilized as an internal standard. Inter-assay variability averaged 5% throughout the assay with γ-aminobutyric acid values in the gerbil hypothalamus of 344 μg/g. The catecholamine assay has been characterized previously and utilizes 3,4-dihydroxybenzylamine as an internal standard with less than 5% variability. Norepinephrine, dopamine, serotonin and 5-hydroxyindoleacetic acid levels in the gerbil hypothalamus averaged 2922, 729, 797 and 272 ng/g, respectively. This new protocol allows a wide range of neurochemicals to be determined and evaluated from the same brain region.