| Background Artemisia argyi Levl.et Vant.is a fragrant and perennial herb of genus Artemisia of family Compositae commonly used for traditional Chinese medicine in Clinical gynecology.This species is rich in terpenoids,flavonoids,phenylpropanoids,aromatic acids（aldehydes）,steroids,and fatty acids with extensive biological activities including of analgesia,hemostasis,antibacterial and antiviral activities,anti-tumor,antimutagenesis,antidiabetics,anti-asthmatic,immunoregulation,antioxidant,and so on.The Traditional Chinese Medicine theory indicates that A.argyi has the efficacy of warming channel.Modern researches display that warming channel is closely related to invigorating the circulation of blood.The preliminary results displayed that the ethyl acetate fraction of A.argyi has an obvious anticoagulant activity.So far,many studies on chemical constituents and biological activity of A.argyi have been reported,but the phytochemical constituents with anticoagulant activities of A.argyi were rarely reported.In order to clarify the material basis of anticoagulant activity of A.argyi and discover the anticoagulant lead compounds,the studies on chemical constituents and anticoagulant activity of ethyl acetate fraction from A.argyi and their anticoagulant activities were carried out.These researches will provide a scientific basis for clarifying the efficacy of warming channel of A.argyi.Objective（1）To investigate the chemical constituents from the ethyl acetate active fraction of A.argyi.（2）To screen anticoagulant activities of the ethyl acetate fraction of A.argyi and some compounds isolated from this fraction.Method（1）Compounds were isolated and purified by various column chromatographic methods including silica gel,Sephadex LH-20 and ODS column chromatographies.Their structures were elucidated on the basis of spectral analysis(IR,UV,1H-NMR,13C-NMR,DEPT,HMBC,HSQC,1H-1H-COSY,NOESY,HRESI-MS),physicochemical methods,and compared with standards.（2）The in vitro anticoagulant activities of the ethyl acetate fraction of A.argyi and ten compounds obtained from this fraction were screened by using the Born turbidimetric method.Result 34 compounds were isolated and identified from the ethyl acetate fraction of A.Argyi,including 17 flavonoids compounds as follows:eupatilin 7-O-β-D-glucopyranoside（1）,5,6,2’,4’-tetrahydroxy-7,5’-dimethoxyflavone（2）,eupatilin（3）,centaureidin（4）,jaceosidin（5）,5,7,4’-trihydroxy-6,3’,5’-trimethoxyflavone（6）,hispidulin（7）,cirsimaritin（8）,5,7,3’-trihydroxy-6,4’,5’-trimethoxyflavone（9）,apigenin（10）,5,3’,4’-trihydrogen-6,7-Dimethoxyflavone（11）,5,7,3’,4’-tetradroxy-6,5’-dihydroxyflavone（12）,luteolin（13）,nepetin（14）,isoquercitrin（15）,daidzein（16）,（S）-5,7,3’,5’-tetrahydroxyflavanone（17）;five sesquiterpenoids:armexifolin（18）,chloroklotzchin（19）,arglanin（20）,ridentin（21）,tanaphillin（22）;one monoterpene:1S,2R,4S-trihydroxyl-p-methane（23）;four phenylpropanoids:umbelliferone（24）,scopoletin（25）,6-methoxycoumarin（26）,caffeic acid（27）;phenolic acid（aldehyde）:o-hydroxybenzoic acid（28）,salicylaldehyde（29）,3,4-dihydroxybenzaldehyde（30）,protocatechuic acid（31）,gentisic acid5-O-β-D-（6’-salicylyl）-glucopyranoside（32）;one sterol:β-sitosterol（33）;one inositol:D-3-O-methyl chiroinositol（34）,respectively.Compounds 1 and 2 are new compounds among them.（2）The anticoagulant activity was screened by the Born turbidimetric method for the ten monomer compounds isolated from the ethyl acetate fraction.The results showed that the ethyl acetate fraction had strong anticoagulant activity.The compounds 1,2,3,5,6,14,15,17,32 of the 10 selected compounds had prominently anticoagulant activity,and compound 32 had the strongest anticoagulant effect.Compounds 5,15,17,32can prolong TT and PT,indicating the anticoagulant effect of them through exogenous and cohesive system.Compounds 1,3,6,and 14 were able to prolong TT,indicating the anticoagulant action via cohesive coagulation.Furthermore,compound 2 significantly prolonged the PT,and compound 7 significantly shorten the TT.Conclusion Two new compounds were isolated from A.argyi.Meanwhile,compounds 6,8,1112,1523,2627,2931,34 were isolated from A.argyi for the first time.The study on anticoagulant activity in vitro showed that compounds 1,2,3,5,6,14,15,17,32 had evident anticoagulant activity compared to blank.This proves that A.argyi actually contain anticoagulant activities compounds. |
