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This Article Full Text Full Text (PDF) All Versions of this Article: clinchem.2005.050062v1 51/9/1698    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (5) Citing Articles via Google Scholar Google Scholar Articles by Mulder, E. J. Articles by Kema, I. P. Search for Related Content PubMed PubMed Citation Articles by Mulder, E. J. Articles by Kema, I. P. Related Collections Automation and Analytical Techniques

Automated On-Line Solid-Phase Extraction Coupled with HPLC for Measurement of 5-Hydroxyindole-3-acetic Acid in Urine

¹ Child and Adolescent Psychiatry, ² Department of Pathology and Laboratory Medicine, and ³ Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
⁴ Child Study Center, Yale University School of Medicine, New Haven, CT.

^aAddress correspondence to this author at: Department of Pathology and Laboratory Medicine, University Medical Center Groningen, PO Box 30.001, 9700 RB Groningen, The Netherlands. E-mail i.p.kema{at}lc.umcg.nl.

Background: 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-hydroxyindole-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.

Conclusion: This SPE-HPLC method demonstrated lower imprecision and time per analysis than the manual solvent extraction method.

The following articles in journals at HighWire Press have cited this article:

				H. Perry and B. Keevil Online extraction of 5-hydroxyindole acetic acid from urine for analysis by liquid chromatography-tandem mass spectrometry Ann Clin Biochem, March 1, 2008; 45(2): 149 - 152. [Abstract] [Full Text] [PDF]

				W. H.A. de Jong, K. S. Graham, J. C. van der Molen, T. P. Links, M. R. Morris, H. A. Ross, E. G.E. de Vries, and I. P. Kema Plasma Free Metanephrine Measurement Using Automated Online Solid-Phase Extraction HPLC Tandem Mass Spectrometry Clin. Chem., September 1, 2007; 53(9): 1684 - 1693. [Abstract] [Full Text] [PDF]