Objective:To establish an HPLC method for the content determination of viterxin in Stenoloma chusana( L. ) Citing. Methods:A Waters Symmetry C18 (250 mm × 4. 6 mm, 5 μm) column was used, and the mobile phase was acetonitrile-0. 1% glacial acetic acid solution (27∶73) at the flow rate of 1. 0 ml·min-1 . The detection wavelength was at 339nm, and the column temperature was 30℃. Results:The linear relationship of viterxin was good within the range from 10. 03 to 90. 27μg·ml-1 . The average recovery was 100. 48%(RSD=0. 98%). Conclusion:The established method is accurate and sensitive with good reproducibility, which can be applied in the determination of viterxin of Stenoloma chusana( L. ) Citing.