| The contents of total flavone and flavonoid monomers in 43 accessions of 13 barrenwort species of genus Epimedium L.,and their chemical fingerprinting,extracting technics and interactive effects with bovine serum albumin were studied,in order to provide some new clues for Chinese medicine modernization.The main results are as follows:(1) The ultraviolet spectrophotometric analysis showed that the average flavone content of 247 plants of 32 accessions in 11 barrenwort species was 6.994%,with the variation coefficient of 44.581%.Among them,E.ecalcaratum has the highest average flavone content(13.166%).E.pubescense,E.pauciflorum,E.wushanense,E. acuminatum,E.chlorandrum and E.myrianthum have average flavone content of 5-10%, and E.pudingense,E.leptorrhizum,E.luodianense,and E.lishihchenii the low average flavone content(less than 5%).This study also showed that there are significant differences of flavone content of E.pubescense and E.acuminatum among different seasons:September＞July＞March＞May.(2) The extraction technics of flavones in E.acuminatum leaves under room temperature has been studied according to the orthogonal design involving in four factors (extracting times,the ethyl alcohol content,the solvent amount,and pH value).The results showed that the extracting times is a key factor to flavone extraction effect. According to the extracting effect of total flavone,the optimum condition is;80%ethyl alcohol solution with 15 time weight more than the raw material,extracting 2 times under non-alkaline condition;As for extracting effect of total icariin,the optimum condition is: 60%ethyl alcohol solution with 20 time weight more than the raw material amount, extracting 2 times under non-alkaline condition.(3)Six flavonoid monomers were isolated from the extract of E.acuminatum by silica gel column chromatography and Sephadex column chromatography.They are identified as Icariin,Icariside-Ⅱ,and 2"-o- rhamnosyl icariside-Ⅱ,4"-o- rhamnosyl icariside-Ⅱ,Anhydroicartin and IkarisosideB by means of ESI MS,1H NMR,and 13C NMR spectra.The others need further studying.(4) The contents of six flavonoid monomers(Icariin,Icariside-Ⅱ,2"-o- rhamnosyl icariside-Ⅱ,4"-o- rhamnosyl icariside-Ⅱ,Ikarisoside B,and Anhydroicartin ) in 40 plants of 13 barrenwort species growing under the same conditions were tested by RP-HPLC. The results indicated that icariin content is relatively high and the others very low.(5)10 common peaks were found in 10 accessions of E.acuminatum from different places of Guizhou Pronvince by HPLC.Among them,the fifth peak is icariin,the seventh peak the is 4"-o- rhamnosyl icariside-Ⅱthe eighth peak is icariside-Ⅱ:the second,the third,and the fourth peaks need further identification.Perhaps they are epimedin A, epimedin B and epimedin C according to the previous reports:the first,the sixth,and the tenth peaks are unknown.(6)In room temperature,2.0 ml solution containing appropriate concentration of BSA was titrated in the Tris-HCl buffer solution(pH7.40) by successive addition of icariin,icariside-Ⅱ,Anhydroicartin and 4"-o- rhamnosyl icariside-Ⅱsolution,respectively. The results indicated that the fluorescence intensities of bovine serum albumin were decreased regularly by the four flavone monomers.
|
PDF Full Text Request