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.
|Cartridge||HySphere Resin GP|
|Compounds||5-Hydoxyindole-3-acetic Acid, 5-HIAA|
|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)|
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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%