OBJECTIVE: To develop a method for simultaneous determination of tenacissoside A and tenacissoside I in Xiaoaiping tablets. METHODS: HPLC method was used to analyze Xiaoaiping tablets after extracted by trichlormethane. The determination was performed in Symmetry C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 223 nm, and the column temperature was 25 ℃. The sample size was 20 μL. The content of tenacissoside A and tenacissoside I in 3 batches of samples were determined by above condition. RESULTS: The linear range of tenacissoside A and tenacissoside I were 26.40-264.00 μg/mL (r=0.999 4) and 4.59-45.90 μg/mL (r=0.999 3), respectively. The limits of detection were 0.26, 0.12 μg/mL, and the limits of quantification were 0.79, 0.36 μg/mL. RSDs of precision, stability (12 h) and repeatability tests were all lower than 2.0%(n=6). The recoveries were 98.53%-99.28% (RSD=0.20%-0.51%,n=9) and 97.00%-98.89% (RSD=0.19%-1.03%,n=9). The contents of tenacissoside A and tenacissoside I in 3 batches of samples were 2.27-2.32 mg/g and 0.64-0.69 mg/g, respectively. CONCLUSIONS: The established method is accurate, simple and reliable, which is suitable for simultaneous determination of tenacissoside A and tenacissoside I in Xiaoaiping tablets.