Determination of hydroxyurea in plasma and peritoneal fluid by high-performance liquid chromatography using electrochemical detection |
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| Affiliation: | 1. Department of Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, Los Angeles, CA 90033, USA;2. Department of Hematology/Oncology and the Kenneth Norris Jr. Comprehensive Cancer Center, School of Medicine, University of Southern California, 1441 Eastlake Avenue, Los Angeles, CA 90033, USA;1. Kazan Federal University, A.M. Butlerov Chemistry Institute, 420008 Kremlevskaya, 18, Kazan, Russian Federation;2. Institute of Fundamental Medicine and Biology, Kazan Federal University, 420008 Kremlevskaya, 18, Kazan, Russian Federation;3. Interdisciplinary Centre for Analytical Microscopy, Kazan Federal University, 420008 Kremlevskaya, 18, Kazan, Russian Federation;4. School of Pharmacy and Biomolecular Sciences, University of Brighton, Huxley Building, Moulsecoomb, Brighton, East Sussex BN2 4GJ, UK;1. Graduate School of Environmental Studies, Tohoku University, 6-6-20, Aramaki, Aoba-ku, Sendai 980-8579, Japan;2. Department of Material Science, University of Shiga Prefecture, 2500 Hassaka-cho, Hikone City, Japan |
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| Abstract: | A sensitive method has been developed for the determination of hydroxyurea in plasma and peritoneal fluid using reversed-phase high-performance liquid chromatography (HPLC) with electrochemical detection. Plasma or peritoneal fluid samples were treated with acetonitrile to precipitate proteins then injected to the HPLC. A C18 analytical column was used to separate hydroxyurea from interfering substances in the biological matrix. The mobile phase, consisting of 0.2 M sodium perchlorate–methanol (95:5, v/v) adjusted to pH 5.0, was delivered isocratically at a flow-rate of 1 ml/min and hydroxyurea was detected using a glassy-carbon electrode operating at an applied potential of +800 mV. Hydroxyurea eluted with a retention time of 3 min. The cycle time for analysis is short and the assay precision is acceptable (C.V. plasma=1.4–3.9%, C.V. peritoneal fluid=2.1–9.7%). The method has been validated and is linear from 25 to 400 ng/ml in plasma and 5 to 30 ng/ml in peritoneal fluid. The method has been shown to be applicable for pharmacokinetic studies. |
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