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Automated On-line Solid Phase Extraction Coupled with HPLC for measurement of 5-Hydoxyindole-3-acetic Acid in urine

Summary Quantification of 5-hydroxyindole-3-acetic acid (5-HIAA) in urine is useful in diagnosing and monitoring of patients with carcinoid tumors and in the study of serotonin (5-hydroxytryptamine) metabolism in various disorders. We describe an automated method that incorporates on-line solid-phase extraction (SPE) and HPLC to measure urinary 5-HIAA. Methods: Automated prepurification of urine was ac­complished with HySphere-resin GP SPE cartridges containing strong hydrophobic polystyrene resin. The analyte (5-HIAA) and internal standard [5-hydroxyin-dole-3-carboxylic acid (5-HICA)] were eluted from the SPE cartridge, separated by reversed-phase HPLC, and detected fluorometrically with a total cycle time of 20 min. Urinary excretion of 5-HIAA was measured in a group of patients with known and suspected carcinoid tumors (n = 63) and in 20 patients with autism.

Results: The internal standard (5-HICA) and 5-HIAA were recovered in high yields (87.2%-114%). Within-and between-series CVs for the measurement of 5-HIAA in urine ranged from 1.2% to 3.9% and 3.2% to 7.6%, respectively. For urine samples from patients with known or suspected carcinoid tumors, results obtained by the automated method were highly correlated (r = 0.988) with those from an established manual extraction method. For samples from autistic patients, urinary excretion of 5-HIAA was similar to that reported for healthy individuals

Category 3. Symbiosis User Application
These are applications that have been made by Symbiosis customers. They are validated and have been published.
Matrix Urine
Market Clinical
Cartridge HySphere Resin GP
Detection HLPC- Fluorescence
Compounds 5-Hydoxyindole-3-acetic Acid, 5-HIAA
Compound Group indoles
Sample_volume 20 ÁL
Contact Erik J. Mulder, Alida Oostenoo-Dmnkerken, George M. Anderson, Elisabeth G.E. De Vries, Ruud B. Minderaa, and Ido P. Kema
Account Department of Pathology and laboratory Medine, UMCG, Groninfgen
Published Clinical Chemistry 51:9, 1698-1703 (2005)
Date 8-8-2005
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Remarks This spe-hplc method demonstrated lower imprececision and time per analysis than the maual solvent extraction method.
recovery yields of 5-HIAA and I.S, are 88-114%
Status published