Colorimetric determination of hydroxyurea in human serum using high- performance liquid chromatography

Konstantine K. Manouilov, Timothy R McGuire, Peter R. Gwilt

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A high-performance liquid chromatography assay for hydroxyurea in human serum was developed based on a commercial colorimetric assay kit for urea (Sigma Diagnostics). Serum (0.5 ml), spiked with methylurea as an internal standard, was treated with 70% perchloric acid. Supernatant (0.2 ml) was combined with 0.7 ml of BUN acid reagent and 0.6 ml of BUN color reagent. The resulting colored reactant (100 μ1) was analyzed on a 300X3.9 mm Bondclone 10 C18 column coupled with a UV-Vis detector, at 449 nm. The mobile phase was 13% acetonitrile in water. Retention times of colored derivatives of hydroxyurea and methylurea were 6.5 and 12.2 min, respectively. The log-log calibration curve was linear from 0.0065 to 1.31 mM. Average accuracy was 99.9±4.0% and the intra- and inter-day error of assay did not exceed 11%.

Original languageEnglish (US)
Pages (from-to)321-324
Number of pages4
JournalJournal of Chromatography B: Biomedical Applications
Issue number1-2
Publication statusPublished - Apr 24 1998



  • Hydroxyurea

ASJC Scopus subject areas

  • Chemistry(all)

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