Determination of 5-hydroxytryptamine, 5-hydroxyindoleacetic acid and tryptophan in plasma and urine by HPLC with fluorimetric detection

Biomedical Chromatography : BMC
C P BearcroftD Perrett

Abstract

Using native fluorescence detection, 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) and tryptophan were resolved from themselves and other naturally occurring compounds using reversed-phase HPLC within 5 min. Deproteinated platelet-poor plasma (PPP) and crude diluted urine were injected directly into the chromatograph. Careful selection of the HPLC column is important and various octadecyl silica (ODS) and base deactivated silic (BDS) columns were evaluated. Pre-treatment of an ODS column with tetrabutylammonium ions gave good selectivity. Between pH 5 and 6 the compounds were well resolved from each other. The limit of quantitative detection of 5-HT and 5-HIAA was 3.5 nmol/L. The overall chromatogram obtained using native fluorescence is cleaner than that obtained with the more commonly employed electrochemical (EC) systems although the chromatography is effectively the same. For analysis of 5-HT in plasma, collection in EDTA was more efficient than lithium heparin. Plasma 5-HT in healthy volunteers was mean 61 (SD = +/- 73) nmol/L, n = 20; urine 5-HIAA gave mean 28.95 (SD = +/- 0.98) mumol/L, (n = 12). Whole blood 5-HT analysis is unreliable in comparison with platelet-poor plasma.

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Citations

Mar 10, 2000·European Journal of Pharmacology·D BurleighD Perrett
Oct 1, 1996·Gut·C P BearcroftM J Farthing
Jan 1, 1997·European Journal of Clinical Chemistry and Clinical Biochemistry : Journal of the Forum of European Clinical Chemistry Societies·A R BonfigliG De Sio
Jan 8, 2005·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Takashi YoshitakeMasatoshi Yamaguchi
Jan 5, 2005·BioFactors·Hirotaka KatsuzakiTakashi Komiya
Jan 31, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Wendy AtkinsonBrian G Keevil
Sep 15, 2005·Environmental Monitoring and Assessment·K SuvardhanP Chiranjeevi
Jun 15, 1997·Analytical Chemistry·D J AndersonK A Davis
Jun 24, 2003·Biochemical Pharmacology·Elena B C LlambíasLeonor C San Martín de Viale
Jun 9, 2021·Scandinavian Journal of Clinical and Laboratory Investigation·Joakim Oxelbark, Annicka Lövenhamn

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