OBJECTIVE： To prepare high-purity rotundic acid， and to determine the contents of syringoside， peduncloside and rotundic acid in Ilicis rotundae and its refined products. METHODS： I. rotundae saponins was prepared by ethanol extraction method and hydrolyzed with methanol and sodium hydroxide； rotundic acid monomer was obtained after recrystallization. The contents of syringoside， peduncloside and rotundic acid in I. rotundae， its refined product 1 and its refined product 2 were determined by RP-HPLC. The determination was performed on an Agilent XDB C18 column with mobile phase consisted of water-acetonitrile （gradient elution） at the flow rate of 1.2 mL/min. The column temperature was 30 ℃， and the detection wavelength was set at 210 nm. The sample size was 10 μL. RESULTS： The purity of rotundic acid exceeded 99％. The linear ranges of syringing， peduncloside and rotundic acid were 0.18-3.22， 0.59-10.36 and 0.20-3.53 μg， respectively （r＝0.999 9）. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2.0％ （n＝6-7）. The average recoveries were 96.08％-100.81％（RSD≤1.98％，n＝6）. The contents of syringoside， peduncloside and rotundic acid in I. rotundae were 1.61％， 7.26％， 1.29％ （RSD≤1.08％， n＝3）； those of refined product 1 were 0， 82.59％， 4.18％ （RSD≤1.67％， n＝3）； those of refined product 2 were 0， 73.29％， 7.41％ （RSD≤1.15％，n＝3）， respectively. CONCLUSIONS： Prepared rotundic acid has high purity and is simple in preparation， safe and environmentally-friendly. RP-HPLC method that established is simple and reliable， and can meet the requirements for rapid analysis of I. rotundae and its refined products.