Study On Extraction Technology And Quality Specification Of Active Components In Xuanmai Ganjie

Objiective:The optimum extraction technology of liquiritigenin,glycyrrhizic acid, Harpagoside and Cinnamic acid in Xuanmai Ganjie was selected with the orthogonal experiment.The content of liquiritigenin,glycyrrhizic acid,Harpagoside and Cinnamic acid in Xuanmai Ganjie Keli was determined by RP-HPLC.In order to control the quality of raw material,semi-product and finished product comprehensively,HPLC fingerprinting of Xuanmai Ganjie Keli was studied.Contents:1.To study the extraction technology of liquiritigenin,glycyrrhizic acid, Harpagoside and Cinnamic acid in raw materials.2.To study the purification technology for the process of production.3.To establish a RP-HPLC method to determinate the contents of the liquiritigenin,glycyrrhizic acid,Harpagoside and Cinnamic acid in Xuanmai Ganjie Keli simultaneously.4.To study the HPLC fingerprinting of Xuanmai Ganjie Keli.Method:To establish RP-HPLC method for the determination of contentof liquiritigenin, glycyrrhizic acid,Harpagoside and Cinnamic acid,diamonds C₁₈column was used as the stationary phase,the mobile phrase was acetonitrile-water(contained 1.0%m-ethanoic acid), detective wavelengthes were 278nm and 254nm.On these grounds,a HPLC fingerprinting of Xuanmai Ganjie Keli was established.Using total content of liquiritigenin,glycyrrhizic acid,Harpagoside and Cinnamic acid as a marker,the optimum extraction technology of raw material was selected with the single factor experiment and orthogonal experiment which selected water volume(A),extraction time(B)and abstraction frequency as examing factors. Xuanmai Ganjie decoction was refined respectively by standing,high speed centrifugalization,alcohol sedimentation,ceramic membrane microfiltration,flocculation. Their refining effect was compared according to indexes of color,clarity,loss rate of active constituents(transfering ratio),and removal ratio of impurity.Result:The optimum extraction process is as follows:adding 24 times water,extraction 3 times,120 min for each time.All kinds of purification made decoction light and clear. Removal ratio of impurity by ceramic membrane microfiltration was the highest and reached 30.33%.Removal ratio of impurity by flocculation was 21.17%,less than that by ceramic membrane microfiltration.Loss rate of active constituents by 50%alcohol sedimentation and ceramic membrane microfiltration were the lowest,and that by flocculation was highest. Good linearities were obtained from 0.21 to 6.30,0.25 to 7.50,0.10 to 3.00 and 0.105 to 3.15μg for liquiritigenin,glycyrrhizic acid,harpagoside and cinnamic acid.The mean recoveries of components were 99.6%(RSD=2.0%),99.3%(RSD=2.5%),100.5%(RSD=2.4%)and 98.9%(RSD=2.2%)respectively.To indicate 20 co-possessing peaks,the relative retention time and relative peak area were obtained.Conclusion:To authenticate the extraction technology five times of raw material according to the optimum technology and draw a conclusion that the result was according with the optimum technology.The extraction technology is stable and feasible.The technique of ceramic membrane microfiltration is optimum purification of Xuanmai Ganjie decoction with the virtues of higher removal ratio of impurity,small amount lose rate of active constituents and simple process.The methods of assaying and finger printing are accurate,rapid and simple and is suitable for full-scale quality control of the Xuanmai Ganjie Keli.