Abstract:To establish a reversed-phase high performance liquid chromatography quantitative method of the determination of osmundancetone in Inonotus obliquus.Different solvents and methods were used to extract Inonotus obliquus.RP-HPLC was applied to determine the content of osmundancetone in the extract of Inonotus obliquus，and the methodology investigation was conducted.The chromatographic conditions were：RP-C18 chromatographic column（150 mm×4.6 mm，5 μm），the column temperature was 25℃，the injection volume was 5 μL，the mobile phase was methanol-0.5%acetic acid water（20∶80），and the flow rate was 0.5 mL/min，the detection wavelength was 320 nm.Osmundancetone showed a good linear relationship with peak area in the range of 0.5 μg/mL-5 μg/mL（r=0.999 9）.The precision，stability and repeatability of the quantitative method were better，and itsRSD were 0.22%，0.77%and 0.59%，respectively.The average recovery rate was 100.05%.In this study，the RP-HPLC quantitative method and pre-treatment process of osmundancetone in Inonotus obliquus were simple and effective.The determination results were accurate，the HPLC method was with good precision and stability，high sensitivity and great repeatability.This method could be used to determine the content of osmundancetone in Inonotus obliquus and other natural product.The study could provide some reference for the development and quality evaluation of inosporium obliquus.