Simultaneous determination of 5-hydroxytryptamine, its amino acid precursors and acid metabolite in discrete brain regions by high-performance liquid chromatography with fluorescence detection.
L-Tryptophan, 5-hydroxytryptophan (5-HTP), 5-hydroxytryptamine, and 5-hydroxyindoleacetic acid (5-HIAA) from discrete regions of brain can be resolved by isocratic elution on a C-18 reversed-phase column and quantitated by native fluorescence measurement. This method is rapid and simple and can reproduceably detect 5-hydroxytryptamine, tryptophan and 5-HIAA with picogram sensitivity. The detection of 5-HTP in brain extracts from untreated animals is difficult because of the extremely low endogenous levels of 5-HTP. Quantitation of 5-HTP presents no problem in animals injected with 5-HTP or with an inhibitor of L-aromatic amino acid decarboxylase. Brain tissue requires minimal treatment for assay and the hydroxyindoles are stable enough to allow automated processing.
D006916:5-Hydroxytryptophan / Q000032:analysis; D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid; D006897:Hydroxyindoleacetic Acid / Q000032:analysis; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012701:Serotonin / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000032:analysis
D000596:Amino Acids; D012701:Serotonin; D006897:Hydroxyindoleacetic Acid; D014364:Tryptophan; D006916:5-Hydroxytryptophan