RESUMO
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.
