Pharmacokinetic Investigation of Main Bioactive Components fromEpimedium-Derived Flavonoids in Rabbit Serum by Ultra-PerformanceLiquid Chromatography Tandem Mass Spectrometry

Background: Total Epimedium-derived flavonoids (EF) were reported to be a group of major constituents present in genus Epimedium for the treatment of osteonecrosis. The present study aimed to investigatethe 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 established and validated by ultra-performance liquid chromatography coupled quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS). Serum samples were pretreated water-saturated ethyl acetate. Furthermore, sample separation 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 mobilephase. Finally, pharmacokinetics of EF in rabbit serum investigated.Results: All calibration curves displayed excellent linearity (r2u003e0.99) within 1.2~24 ng/mL. The assay was specific, precise and accurate, as demonstrated with intra-run 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 successfully applied to analyse thepharmacokinetic 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 thetreatment of osteonecrosis in rabbit.