Introduction: In this study, a precise, rapid and accurate ultra-performance liquid chromatography-
tandem mass spectrometry (UPLC-MS/MS) method for the quantification of zhebeirine in
mouse blood was developed, and pharmacokinetics of zhebeirine was studied for the first time after
intravenous and oral administration.
Methods: The mobile phase consisted of acetonitrile-0.1% formic acid, with a flow rate at 0.4 mL/m
during 4 min run time. MRM modes of m/z 414.5→81.0 for zhebeirine and m/z 430.2→412.2 for 3-
dehydroverticine (internal standard) were utilized to perform quantitative analysis. Protein in mouse
blood was directly precipitated with acetonitrile for sample preparation.
Results: The linear range was 1-3000 ng/mL with r>0.995, and LLOQ was 1 ng/mL. The intra-and
inter-day precision of zhebeirine in mouse blood was less than 13%. The accuracy ranged from 91.2%
to 112.5%, while the matrix effects were between 84.8% and 106.4%.
Conclusion: The UPLC-MS/MS was successfully applied to the pharmacokinetic study on zhebeirine
after intravenous and oral administration, and the bioavailability was determined to be 22.8%.