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.
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.
Yoshitake
Takashi
T
Chemicals Evaluation and Research Institute, Japan, 3-822 Ishii-Machi, Hita, Oita 877-0061, Japan.
Iizuka
Ryuji
R
Fujino
Kaoru
K
Inoue
Osamu
O
Yamagata
Kenji
K
Nohta
Hitoshi
H
Yamaguchi
Masatoshi
M
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
D012701:Serotonin; D006897:Hydroxyindoleacetic Acid