Analytical Sciences
Online ISSN : 1348-2246
Print ISSN : 0910-6340
ISSN-L : 0910-6340
Liquid Chromatographic Determination of Indoleamine, Catecholamines and Their Precursors and Metabolites in Rat Brain Tissues with Postcolumn Fluorescence Derivatization
Hitoshi NOHTAHee-Kyoung JEONMasaaki KAIYosuke OHKURA
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1994 Volume 10 Issue 1 Pages 5-9

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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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© The Japan Society for Analytical Chemistry
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