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

Journal of Chromatography. B, Biomedical Sciences and Applications
K K ManouilovP R Gwilt

Abstract

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 microl) was analyzed on a 300 x 3.9 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%.

References

Dec 23, 1992·Journal of Chromatography·J HavardD Sampson

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Citations

Sep 16, 2003·Journal of Clinical Pharmacology·Peter R GwiltSusan S Swindells
Oct 13, 2009·Fundamental & Clinical Pharmacology·Hind SassiAnne Hulin
Dec 28, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hutchinson JamesRobert E Taylor
Dec 20, 2005·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·David A ReardonHenry S Friedman
May 15, 2015·Therapeutic Drug Monitoring·Uttam GargKathleen Neville
Jul 6, 2018·Frontiers in Chemistry·Roberta Risoluti, Stefano Materazzi

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