Rapid determination of p-aminohippuric acid in serum and urine by high-performance liquid chromatography

R. Marsilio, R. Dall'Amico, G. Montini, L. Murer, M. Ros, G. Zacchello, F. Zacchello

Research output: Contribution to journalArticle

Abstract

We report a simple and rapid HPLC procedure for measuring p-aminohippuric acid in serum and urine. After deproteinization with acetonitrile and addition of p-aminobenzoic acid as an internal standard, the chromatographic run is performed on a C18 column with the absorbance detector set at 275 nm. The separation is carried out in 10 min at a flow-rate of 1.0 ml/min with a mobile phase composed of 7 mmol/l l-decanesulfonic acid, pH 3.70, and acetonitrile (82:18, v/v). The relationship between p-aminohippuric acid concentration and the p-aminohippuric acid/internal standard peak area is linear up to 100 μg/ml. Within-run precision measured at three different p-aminohippuric acid concentrations ranges from 1.73 to 1.98% in serum and from 0.72 to 1.32% in urine. Between-run precision varies from 1.80 to 3.06% in serum and from 1.05 to 2.66% in urine. Analytical recovery is between 98.26 and 99.44% in serum and from 99.57 to 100.45% in urine. The method is very simple, sensitive, and requires small volumes of sample for the assay (100 μl). Therefore, it could be a useful tool for the analysis of p-aminohippuric acid in the evaluation of renal plasma flow.

Original languageEnglish
Pages (from-to)359-364
Number of pages6
JournalJournal of Chromatography B: Biomedical Applications
Volume704
Issue number1-2
DOIs
Publication statusPublished - Dec 19 1997

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Keywords

  • p-Aminohippuric acid

ASJC Scopus subject areas

  • Chemistry(all)

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