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

Determination of 5‐hydroxytryptamine, 5‐hydroxyindoleacetic acid and tryptophan in plasma and... Using native fluorescence detection, 5‐hydroxytryptamine (5‐HT), 5‐hydroxyindoleacetic acid (5‐HIAA) and tryptophan were resoved 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 pH5 and 6 the compounds were well resolved from each other. The limit of quantitatiave detection of 5‐HT and 5‐HIAA was 3.5 nmol/L. The overal 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)μmol/L, (n=12). Whole blood 5‐HT analysis is unreliable in comparison with platelet‐poor plasma. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biomedical Chromatography Wiley

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

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References (11)

Publisher
Wiley
Copyright
Copyright © 1995 John Wiley & Sons, Ltd.
ISSN
0269-3879
eISSN
1099-0801
DOI
10.1002/bmc.1130090105
pmid
7537559
Publisher site
See Article on Publisher Site

Abstract

Using native fluorescence detection, 5‐hydroxytryptamine (5‐HT), 5‐hydroxyindoleacetic acid (5‐HIAA) and tryptophan were resoved 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 pH5 and 6 the compounds were well resolved from each other. The limit of quantitatiave detection of 5‐HT and 5‐HIAA was 3.5 nmol/L. The overal 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)μmol/L, (n=12). Whole blood 5‐HT analysis is unreliable in comparison with platelet‐poor plasma.

Journal

Biomedical ChromatographyWiley

Published: Jan 1, 1995

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