Abstract
Background: Total Epimedium-derived flavonoids (EF) were reported to be a group of major bioactive constituents present in genus Epimedium for the treatment of osteonecrosis. The present study aimed to investigate the pharmacokinetics of main bioactive components of EF in rabbit. Methods: A rapid and sensitive method for the simultaneous determination of icariin, icariside I, icariside II and icaritin in serum was established and validated by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS). Serum samples were pretreated with water-saturated ethyl acetate. Furthermore, sample separation was achieved on an Waters BEH C18 column (50 mm × 2.1 mm, 1.7 μm) by gradient elution with acetonitrile-water (both including 0.1% formic acid) as mobile phase. Finally, pharmacokinetics of EF in rabbit serum was investigated. Results: All calibration curves displayed excellent linearity (r2>0.99) within 1.2~24 ng/mL. The assay was specific, precise and accurate, as demonstrated with intra-run precisions not more than 12.6%, inter-run precisions less than 9.9% and accuracies between -4.4% and 9.3%. Meanwhile, the matrix effects, extraction recoveries and stabilities were all satisfactory. Moreover, this validated method was successfully applied to analyse the pharmacokinetic of EF in rabbit serum after oral dosage of 80 mg/kg. Conclusion: These results would enlarge our knowledge about main bioactive components of EF for the treatment of osteonecrosis in rabbit.
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