Separation Process Of Three Phenolic Glycosides From Monochasma Savatieri Franch And Antiinflamation Activity Research

Monochasma savatieri Franch(Scrophulariaceae)is also called "Longxucao","Baitouweng",etc,,which is widely distributed in Jiangsu,Zhejiang,Hunan and Jiangxi provinces of China.As a famous traditional Chinese medicine,the whole plant is extensively used for the treatment of cough,cold and inflammation.It is one of the main components of Yanning granula,which has been collected into protection of the traditional Chinese medicine species.In our recent study,12 phenylethanoid glycosides were isolated,and the total phenylethanoid glycosides(TPG)from Monochasma savatieri exhibited strong cardiovascular activity,antibacterial and anti-inflammatory activity.In order to explore the bioactive agents and accumulate the quantity for further biological research,the preparative technology of TPG from Monochasma savatieri Franch was studied and discussed in this paper,as well as the anti-inflammatory activity of Acteoside,Isoacteoside and Torenoside B.In this thesis,a RP-HPLC method was established for the simultaneous determination of Acteoside,Isoacteoside and Torenoside B of Monochasma savatieri Franch,and the results indicated the linear ranges of Acteoside,Isoacteoside and Torenoside B were 0.59?4.51?g(R2=0.9999),0.61?4.65?g(R2=0.9997),0.54?4.66?g(R2=0.9998),respectively.And the average recoveries of Acteoside,Isoacteoside and Torenoside B were 100.20%?97.92%and 97.18%,respectively.This method is simple,reliable and accurate for determination of Acteoside,Isoacteoside and Torenoside B and in antibacterial active fraction of Monochasma savatieri Franch.Eight resins(AB-8,D101,HPD100,LSA-10,LX-11,LX-17,LX-38,and XDA-6)were used to investigate their static adsorbed capacity and desorbed capacity through the pertinent static experiments.Acteoside was adsorpted 25.85 and 25.63 mg/g on LX-17 and LX-38,respectively.Isoacteoside was adsorpted 43.38 and 34.60 mg/g and Torenoside B was adsorbed 31.32 and 30.96 mg/g.The rates were all over 90%on LX-17 and LX-38.The data indicated that LX-17 and LX-38 were optimal resins for further experiments comparing with other resins.The dynamic experiments indicated that adsorbed contents and desorption rates on LX-17 were better than the ones on LX-38.Therefore,LX-17 was chosen to investigate the related optimal parameters.The adsorption isotherms experiments indicated that the optimal feeding concetration of ram materials was 1.8 g/mL.The adsorption kinetic experiments demonstrated that the optimal adsorption time was 210 min.The dynamic desorption curves indicated that 15%ethanol could not desorb over 5%samples,and 80%ethanol could efficiently desorb samples.Therefore,the eluent process was designed to 0%,15%,80%and 95%ethanol.And then the scale-up enrichment of Acteoside,Isoacteoside and Torenoside B experiments was performed in our laboratory using the optimal parameters aforementioned.The total phenylethanoid glycosides obtained from scale-up experiments was further separated by preparative semi-HPLC with dynamic axial compress system to simultaneously purify Acteoside,Isoacteoside and Torenoside B.In this thesis,the inflammatory mode with RAW264.7 macrophage line induced by LPS was established.The nitrite level of RAW264.7 was measured using Griess aasay.Our data indicated that the inhibitory effects were significant on the NO of RAW264.7 cells line treated with 40 and 20 ?M Acteoside,Isoacteoside and Torenoside B,respectively.However,10?M Acteoside,Isoacteoside and Torenoside B showed no significance.The proliferation effects of Acteoside,Isoacteoside and Torenoside B on RAW264.7 indicated that the indicated various concentration showed no effect on the growth.