Abstract: OBJECTIVE To establish a method for plasma concentration of decursin in rat plasma by UPLC-MS/MS and apply to the pharmacokinetic characteristics of decursin after oral and intravenous injection. METHODS The protein of rat plasma was removed by acetonitrile precipitation method. Diazepam was used as internal standard. UPLC BEH C₁₈ column (2.1 mm×50 mm, 1.7 μm) was used as the separation column. Acetonitrile and 0.1% formic acid was used as a mobile phase by gradient elution. The flow rate was 0.4 mL·min^-1, the analysis time was 4.0 min. The quantitative analysis were detected by electrospray ionization(ESI), positive ion detection combined with multiple reaction monitoring mode. The following transitions were obtained at m/z 329.0→229.0 for decursin and m/z 285.1→193.3 for internal standard. RESULTS The linear relationship of decursin in 1-60 ng·mL^-1 was good and limit of quantitation was 1 ng·mL^-1. Intra-day and inter-day precisions were <14%, the accuracy range was between 88.5% and 107.5%, the recovery was >93.3%, and the matrix effect was between 89.3% and 93.5%. CONCLUSION The method is sensitive, fast, specific, and has been successfully applied to a pharmacokinetic study of decursin.