An automated method for the determination of biogenic amines and their metabolites by high-performance liquid...

An automated high-performance liquid chromatographic method has been developed that allows for the determination of a number of compounds related to catechol- and indoleamine metabolism. The compounds that can be measured include L-DOPA, dopamine, dihydroxyphenylacetic acid, homovanillic acid, 3-methoxytyramine, norepinephrine, 3-methoxy-4-hydroxyphenylglycol, dihydroxyphenylglycol, vanilmandelic acid, epinephrine, 5-hydroxytryptophan, serotonin, 5-hydroxyindoleacetic acid, and 5-hydroxytryptophol. Dihydroxybenzylamine is used as an internal standard. Although dihydroxyphenylglycol and vanilmandelic acid could be detected and quantified, they could not be separated from each other. The method is completely automated and is sensitive enough to detect amounts as low as 500 fmol. Up to 200 samples a week can be analyzed in the automated mode. Using this method, analyses of brain tissue can be accomplished with no need for a cleanup procedure. The value of this procedure lies in its ability to simultaneously determine various amines and metabolites from small tissue samples in the same animals and with automation to analyze a relatively large number of samples a day with little attention by a technician.