Objective: To investigate in vitro transdermal absorption properties of Dingguier umbilical pastes. Method: Vertical Franz diffusing cells and nude mice were adopted for percutaneous penetration test.HPLC was adopted to determine accumulative percutaneous permeation of eugenol, cinnamic aldehyde and piperine in certain time with mobile phase of methanol (A)-water (B) gradient elution (0-25 min, 50%-90%A) and detection wavelength of 290, 280, 343 nm, respectively. Result: Linear ranges of eugenol, cinnamic aldehyde and piperine were 0.0144 6-2.892 5, 0.001 1-0.226 2, 0.002 0-0.403 2 μg, average recoveries were 100.5%, 98.6%, 99.1%, transdermal time-lags were 0.07, 0.41, 2.36 h, penetration rates were 49.91, 2.65, 0.40 μg·cm^-2·h^-1, accumulative penetration amounts in 24 h were (1 204.28±103.74), (63.01±10.95), (8.82±1.19) μg·cm^-2, respectively. Conclusion: In vitro percutaneous penetration of Dingguier umbilical pastes complied with zero-order kinetics.