title: Simultaneous determination of serotonin and 5-hydroxyindole-3-acetic acid in human urine by automated precolumn derivatization and semi-microbore column liquid chromatography with fluorescence detection.
abstract: An improved high-performance liquid-chromatographic (HPLC) assay for the simultaneous determination of serotonin (5-HT) and 5-hydroxyindole-3-acetic acid (5-HIAA) in human urine is reported. Following the automated precolumn derivatization of urinary 5-HT and 5-HIAA with benzylamine, the derivatives are separated by isocratic elution on a reversed-phase C18 semi-microbore column, and are fluorometrically detected at an excitation wavelength of 345 nm and an emission wavelength of 480 nm. The detection limits of 5-HT and 5-HIAA are 7 and 5 nmol/l in urine (0.7 and 0.5 fmol/20-microl injection). The proposed automatic method permits a highly selective and sensitive determination of 5-HT and 5-HIAA in human urine without any sample purification.
authors.affiliation: Chemicals Evaluation and Research Institute, Japan, 3-822 Ishii-Machi, Hita, Oita 877-0061, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yoshitake
authors.forename: Ryuji
authors.initials: R
authors.lastname: Iizuka
authors.forename: Kaoru
authors.initials: K
authors.lastname: Fujino
authors.forename: Osamu
authors.initials: O
authors.lastname: Inoue
authors.forename: Kenji
authors.initials: K
authors.lastname: Yamagata
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Nohta
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Yamaguchi
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D005453:Fluorescence; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000652:urine*; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000652:urine*; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D012701:Serotonin; D006897:Hydroxyindoleacetic Acid