Abstract: Salicin, Salicyloylsalicin, and Leteolin 7-O-glucoside were determined from S. Oritrepha. Methods: High-performance liquid chromatography(HPLC) on a Symmetry C₁₈ column (4.6 mm×250 mm, 5 μm) with acetic acid-methanol gradient elution, at column temperature of 30℃, flow-rate of 1.1 mL/min and wavelength at 270 nm. Results: The contents of three compounds can be determined simultaneously from S. Oritrepha.The linear concentration ranges of these compounds were 527.0～105.4 μg/mL, 140.0～28.0 μg/mL, 81.0～16.2 μg/mL respectively. The average recoveries were 104.9%, 104.6% and 103.6% respectively, and relative standard deviations were 2.0%, 0.3% and 0.3% respectively. Conclusion: The proposed method was a simple and rapid method for simultaneous determination of the three mentioned compounds in various tissues from S. Oriterpha.