3,4-Dimethoxybenzylamine as a sensitive pre-column fluorescence derivatization reagent for the determination of serotonin in human platelet-poor plasma.
3,4-Dimethoxybenzylamine is shown to be a highly sensitive pre-column fluorescence derivatization reagent for the determination of serotonin in plasma by high-performance liquid chromatography. The reagent reacts selectively with 5-hydroxyindoles including serotonin in slightly alkaline media in the presence of potassium hexacyanoferrate(III) to give highly fluorescent derivatives. The derivatives of six standard 5-hydroxyindoles (5-hydroxytryptophan, 5-hydroxyindole-3-acetic acid, serotonin, 5-hydroxyindole-3-acetamide, N-acetyl-5-hydroxytryptamine and 5-hydroxytryptophol) are separated within 18 min by isocratic elution using acetonitrile-10 mM phosphate buffer (pH 6.0)-50 mM 1-hexanesulfonic acid on a Wakosil II 5C18RS reversed-phase column. The detection limits (signal-to-noise ratio=3) for the indoles were 1.0-5.7 fmol in a 100-microl injection volume. The method was applied to the measurement of serotonin in human platelet-poor plasma.
Ishida
J
J
Faculty of Pharmaceutical Sciences, Fukuoka University, Johnan-ku, Japan.
Takada
M
M
Yamaguchi
M
M
D001596:Benzylamines; D001792:Blood Platelets; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000097:blood*; D013050:Spectrometry, Fluorescence
D001596:Benzylamines; D005456:Fluorescent Dyes; D012701:Serotonin; C069986:3,4-dimethoxybenzylamine