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Clinical Chemistry 26: 910-912, 1980;
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Clinical Chemistry, Vol 26, 910-912, Copyright © 1980 by American Association for Clinical Chemistry

Liquid-chromatographic separation of urinary 5-hydroxy-3-indoleacetic acid, with measurement at 254 nm

PS Draganac, SJ Steindel and WG Trawick

A "high-performance" liquid-chromatographic procedure for 5-hydroxy-3- indoleacetic acid is described and compared with a colorimetric method in which 1-nitroso-2-naphthol is used. The analyte and an internal standard, p-nitrobenzoic acid, were extracted into diethyl ether from urine at pH 4.0 (acidified with HCl) to which sodium chloride had been added, and the ether was back-extracted with acetate buffer, pH 9.2. Aliquots of this extract were injected into a reversed-phase liquid- chromatographic column and eluted with pH 3.5 acetate buffer/methanol (95/5 by vol); the effluent was monitored at 254 nm. The precision (CV) of the method was 11.8% at 1.8 mg/L, 5.5% at 92 mg/L. Analytical recovery averaged 84%. The colorimetric method gave higher values for the analyte than did the chromatographic method for all patients' urines.


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


Home page
 Ann Clin BiochemHome page
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]


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Copyright © 1980 by the American Association for Clinical Chemistry.