Abstract: To set up a determination method and determine the contents of aucubin in Pedicularis henryi Maxim using HPLC.The conditions of Chromatography:Waters exterra RP18 column was used with a mobile phase of MeOH-H₂O (2∶98,v/v) at flow rate:0.5 mL/min,to determine aucubin in the sample with wavelength 205 nm.The calibration curve of aucubin was in good linearity over the range of 0.02—1 μg,The average recovery were 101.83%,the relative standard deviations(RSD) were 4.25%.The method is reproducible,accurate and reliable; through the determination of the content of aucubin from different places Pedicularis henryi,provides the reference for the quality control of Pedicularis henryi.