Analysis for indole compounds in urine by high-performance liquid chromatography with fluorometric detection.
We describe a chromatographic system involving a high-performance chemically-bonded reverse-phase column and fluorescence detection for measurement of indoles in urine. We controlled retention and selectivity by optimizing the methanol content and pH of the mobile phase. Six reference indoles were separated in less than 20 min; three 5-hydroxyindoles were eluted in less than 7 min. About 5-15 ng of aqueous solutions of these compounds can be detected. The combination of selectivity (from use of the chromatographic column) and fluorescence detection permitted analysis for five of the six indoles after a single urine-deproteinization step.
D006916:5-Hydroxytryptophan / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005069:Evaluation Studies as Topic; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006897:Hydroxyindoleacetic Acid / Q000652:urine; D007210:Indoleacetic Acids / Q000652:urine; D007211:Indoles / Q000652:urine*; D012701:Serotonin / Q000652:urine; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014363:Tryptamines / Q000652:urine; D014364:Tryptophan / Q000652:urine
D007210:Indoleacetic Acids; D007211:Indoles; D014363:Tryptamines; D012701:Serotonin; D006897:Hydroxyindoleacetic Acid; D014364:Tryptophan; D006916:5-Hydroxytryptophan