DEVELOPMENT AND VALIDATION OF A RAPID AND SIMPLE REVERSED-PHASE HPLC METHOD FOR THE DETERMINATION OF GEMCITABINE IN HUMAN PLASMA
Keywords:Gemcitabine, Cytarabine, Reversed-phase HPLC, Plasma
AbstractObjective: In order to investigate the human plasma pharmacokinetics of dFdC, the objective of this work was to optimize and validate a rapid reversed-phase (RP) high-performance liquid chromatography (HPLC) method according to the guidelines of the international regulatory institutions: European Medicines Agency (EMA), Food and Drug Administration (FDA) and International Conference on Harmonization (ICH). Methods: Chromatographic runs were performed on a RP-ACE-C18 column. Mobile phase was constituted of sodium acetate buffer (pH 5) and acetonitrile, in gradient mode, at a flow rate of 1 mL/min. Gemcitabine and cytarabine (internal standard) were detected at 290 nm. Results: The method was shown to be selective, linear in the range of 0.25â€“10 mg/L (R2=0.9998), accurate and precise within-run and between-run as reflected by the coefficient of variation values (<15%) and the relative errors values (<15%), stable and robust to changes in the column temperature and detection UV wavelength. Detection limit and lower limit of quantification were 0.22 and 0.25 mg/L respectively. Conclusion: The developed method is useful to measuring gemcitabine plasmatic concentrations in pharmacokinetics studies and in therapeutic drug monitoring.
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