Abstract
This paper describes a method for the determination of indoleamine (5-hydroxytryptamine) and its precursor (5-hydroxytryptophan) and metabolites (5-hydroxyindoleacetic acid and 5-hydroxytryptophol) simultaneously with catecholamines (norepinephrine, epinephrine and dopamine) and their precursor (L-3, 4-dihydroxyphenylalanine) and metabolites (normetanephrine, metanephrine, 3-methoxytyramine, 3, 4-dihydroxymandelic acid, 3, 4- dihydroxyphenylacetic acid, vanillylmandelic acid, homovanillic acid, 3, 4-dihydroxyphenylethylene glycol and 4-hydroxy-3-methoxyphenylethylene glycol). The method uses reversed-phase liquid chromatography with postcolumn fluorescence derivatization involving a periodate oxidation. This is followed by reaction with meso-1, 2-diphenylethylenediamine in the presence of hexacyanoferrate(III). The detection limits (S/N=3) varied from 13 to 570fmol on column, depending on the compounds. The method can be applied to the determination of these compounds in four discrete brain regions (striatum, cerebral cortex, hypothalamus and hippocampus) from rats treated with saline, pargyline (a monoamine oxidase inhibitor) and probenecid (an acid transport inhibitor).
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