Extraction Of The Effective Site Of Callicarpa Kwangtungensis Chun Phenylethanol Glycoside And Study On Chemical Composition

Callicarpa kwangtungensis Chun,is widely distributed in the province of Jiangxi,Guangdong and other places in China,which has the effects of astringent,hemostasis,heat clearing and detoxification.It is the main raw material of traditional Chinese patent medicine such as "KanggongyanPian" and " KangGongYanJiaoNang".The chemical composition of Callicarpa kwangtungensis Chun contains flavonoids,phenylethanoid glycosides,phenylpropanoid,triterpenoids,diterpenoids,sesquiterpenoids and phenolic acids,and has been confirmed that phenylethanoid glycosides are one of the most importent effective components,which has attracted much attention.At present,there are relatively few studies on Callicarpa kwangtungensis Chun at home and abroad,and the material basis of its efficacy is not clear enough.In order to develop and utilize the traditional Chinese medicine competely,we selected the research object of Callicarpa kwangtungensis Chun to carry out the following research:1.Study on the extraction technology of phenylethanoid glycosides from Callicarpa kwangtungensis Chun.Three methods of heating reflux,ultrasonic extraction and cold extraction were investigated.Finally,the heating reflux extraction method was selected.The single factor test was used to investigate the influence of five factors:ethanol concentration,extraction time,solid-liquid ratio,extraction temperature and extraction times on the yield of phenylethanoid glycosides.The optimal process was finally determined as follows:ethanol concentration 60%,solid-liquid ratio 1:15,reflux extraction 3 times,1.5 hours each time.The yield of phenylethanol glycoside can reach 1.06%.2.Study on the enrichment process of phenylethanoid glycosides from Callicarpa kwangtungensis Chun.Through the study of the static adsorption characteristics of three macroporous adsorption resins,it was determined that X-5 macroporous adsorbent resin was an ideal macroporous adsorbent resin and suitable for the separation and purification of phenylethanol glycoside from Callicarpa kwangtungensis Chun.Through the study on the dynamic adsorption characteristics of X-5 macroporous adsorbent resin,the optimum process was determined as:the loading volume (857 μg/g),loading concentration (8 mg/mL),loading volume flow 1 BV/h,elution volume fraction 40% ethanol,elution volume flow 1 BV/h and elution solvent amount 5 BV.3.Study on the chemical composition of Callicarpa kwangtungensis Chun and hemostatic activity.Two new phenylethanoid glycosides,3-O-α-L-rhamnopyranosyl-6-O-D-apiofuranosyl-4-O-E-caffeoyl-D-glucopyranoside(A-1*) and 3,6-O-α-L-dirhamnopyranosyl-4-O-E-caffeoyl-D-glucopyranoside (A-2*),along with thirteen known ones (from A-3 to A-15),were isolated from hemostatic active fraction of Callicarpa kwangtungensis by using various column chromatography,including MCI gel,ODS,Sephadex LH-20,and semi-preparative HPLC.Their structures were determined on the basis of spectroscopic analysis,and comparison with those in literature.In addition,the NMR data of compounds A-3 and A-4 were reported here for the first time.Compared with Yunnan Baiyao as the positive control drug,the 60% ethanol extract and naturally abundant ingredients A-12 and A-15 could significantly shorten bleeding time of mice in the tail tip bleeding model.