High-performance liquid chromatographic determination of 5-hydroxyindoles by post-column fluorescence derivatization.
A selective and sensitive high-performance liquid chromatographic method with post-column fluorescence detection has been developed for the determination of 5-hydroxyindoles. Five biogenic 5-hydroxyindoles (5-hydroxytryptophan, 5-hydroxytryptamine, 5-hydroxyindol-3-ylacetic acid, 5-hydroxytryptophol and N-acetyl-5-hydroxytryptamine) were separated by isocratic elution on a reversed-phase column, TSKgel ODS-80Tm, and then converted into fluorescent derivatives by post-column reaction with benzylamine and potassium hexacyanoferrate(III) reagents. The detection limits (signal-to-noise ratio = 3) for the indoles were in the range 140-470 fmol per injection volume (100 microliters). The method was applied to the determination of 5-hydroxyindol-3-ylacetic acid in human urine, with direct sample injection.
Ishida
J
J
Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
Iizuka
R
R
Yamaguchi
M
M
D000328:Adult; D001596:Benzylamines; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D005453:Fluorescence; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000652:urine; D007202:Indicators and Reagents; D007211:Indoles / Q000032:analysis* / Q000652:urine; D008297:Male; D012016:Reference Values
D001596:Benzylamines; D007202:Indicators and Reagents; D007211:Indoles; D006897:Hydroxyindoleacetic Acid; C030796:benzylamine