Preparation And Application Of Reference Extracts From Wine-prepared Fructus Corni

Objective:To study the preparation and application of wine-prepared Fructus Corni reference extract.Methods:The single factor investigation and orthogonal design(with L9(34)orthogon-al table)was used to optimize the extraction process of Morroniside,verbenalin,sweroside and loganin,three factors in extraction were optimized.By comparing the content and transfer ratio during the process of purification,we optimized the process of column chromatography of macroporous resin and Aluminum Oxide.Using wine-prepared Fructus Corni reference extract instead of standard determi-nation of the contents of Morroniside,verbenalin,sweroside and loganin from 15 batch wine-prepared Fructus Corni.Rusults:The extract condition of reference extract of wine-prepared Fructus Corni was as follows: wine-prepared Fructus Corni was heating reflux extracted with10 folds of distill water for two times,2 hours each time.The optimal process for purification of morofficianloside with HPD400 macroporous absorption resin was as follows:the concentration of the extract fluid of wine-prepared Fructus corni 2:1,the flow rate 1BV/h(1BV represented one column volume),The saturation of HPD-400 is 2.5 g(crude drug)/g(macroporous resins);firstly eluting the water-soluble impurity like amylose by distilled water,then eluting by 30% alcohol,collecting the part eluted by 30% alcohol;the eluting cubage of 30% alcohol 2.5BV,the eluting rate 2BV/h.The best neutral aluminium oxide(100~200 mesh)column chromatographic methods are: the amount of the sample weight alumina 1/30,mobile phase(water/alcohol 5∶95 V/V),the eluting cubage of 95% alcohol 17.5BV,the purity of reference extract can be improved to more than 80% after neutral alumina column chromatographic separation.The alternative raw materials have good uniformity and stability,experimental results show that the stability of thewine-prepared Fructus Corni reference extract hashygroscopic,high temperature and strong light resistance,should be sealed at room temperature.The marked values of four components have been determined through the statistical analysis of the determining data(Morroniside 41.7%,verbenalin 3.67%,sweroside 2.07% and loganin 32.73%).Using wine-prepared Fructus Corni reference extract instead of standard determination of the contents of Morroniside,verbenalin,sweroside and loganin from15 batch wine-prepared Fructus Corni.And proved the feasibility of the method is simple,has a good reproducibility and accuracy.Conclusion:The preparation process of the reference extract from wine-prepared Fructus Corni presents a good stability and reproducibility.Therefore,it can provide a reference for the quality control of wine-prepared Fructus Corni.