Study on the separation and purification of ursolic acid from Ilex kudingcha

OBJECTIVE The separation and purification of ursolic acid from Ilex kudingcha C.J.Tseng was studied in order to exploit its resource.METHODS The effect of different factors on leaching rate was studied by orthogonal experiment and variance analysis. Ursolic acid was obtained by extraction, precipitation and macro-porous resin adsorption. The samples were analyzed by HPLC. The mobile phase consisted of alcohol and water, and the eluate was collected, in which, ursolic acid was detected at 220 nm. RESULTS The optimal conditions were obtained.The leaching rate of ursolic acid was up to 1.44% by reflux extraction using 90% alcohol for 2.5 h twice at 1:14 of Ilex kudingcha C. J. Tseng mass to solvent volume. The optimal conditions of separation and purification of D8 resin included: the ursolic acid concentration was 1-2 mg·mL-1 and the solution was pH 6.0-7.0, eluate was pH 11.0. After adsorpted for an hour, resin was eluted successively with water, four times volume 30% alcohol,four times volume 90% alcohol in the flow rate of 2-3 mL·min-1. Ursolic acid product was acquired from 90% alcohol eluate.The purity of product was over 95% and the recovery was up to 97.82% . CONCLUSION The product was obtained with high purity and high recovery. It can be utilized in industrial production.