Wushan Icariin Preliminary Study On Chemical Constituents And Wine Icariin

As a group of traditional chinese herbs owning a long history and first recorded in Shennong's Canon of Herbal Materia Medica, Epimedium (Herba Epimedii) are characterized by bitter and acrid in taste and cold in nature, having significant effects on tonifying the liver and kidney,strengthening the bones and muscles,expelling wind and removing dampness. 5 varieties of Epimediums has been recorded in Chinese Pharmacopoeia (2005 Edition). By comparison to others, E.wushanense T.S.Ying is relatively easy to be distinguished: With hard and crispy texture of leaves, it has narrow-long blade profile, whose length is as long as 6 times than the width, and 7 vascular bundles in its midrib. Meanwhile, with broad cultivation planting bases, this one is used as the mainstream variety in clinical practice.However E.wushanense T.S.Ying is widely in use and has significant medical effects, its content of Icariin is far below the Pharmacopoeia standard. Under such circumstances, it is of great necessity to study its chemical ingredients, which has not been appropriately studied yet. Through this research, it is expected to choose some proper chemical compositions as its quality control index and to isolated such compounds as its material foundation, on the basis of which to carry on studies on processed Epimedium.In addition, famous rare books in past dynasties have many records about wine processed E.wushanense T.S.Ying which was mentioned in dozens of books and literatures used extensively as a medicine, only second to the one processed by mutton oil. But this class of E.wushanense T.S.Ying products was rarely used any longer and was replaced in principle. To avoid abandoning the valuable experience of our predecessors, it should be after considerate research on the wine processed E.wushanense T.S.Ying before making the conclusion whether it is reasonable to reject this wine processing technology and having clinical effects or not. On the other hand, because of the little research on the the processing mechanism of Epimedium, it is of great importance to do further study on it.Therefore, based on the reasons mentioned above, this research was carried out from the following aspects:1. studies on the chemical constituents of E.wushanense T.S.Ying: proper solvent method was applied in the extraction of E.wushanense T.S.Ying, whose chemical components was isolated and identified by integrating several chromatographic separation methods and modern spectral analysis methods, and 11 chemical compounds were obtained, among which 8 compounds was recognized. And 6 out of these 8 were flavonoids, which are Epimedin A isolated from E.wushanense T.S.Ying for the first time,Epimedin B,Epimedin C,Icariin,IcarisideⅠand IcarisideⅡ, besides the 6 mentioned above, the other 2 wereβ-sitosterol and daucosterol. The content of Epimedin C in E.wushanense T.S.Ying is quite high, even higher than that of Icariin, and with similar chemical structure, Epimedium C has just one more rhamnose than Icariin. Therefore, with so many similarities to each other, it is suggested that Epimedin C might have similar medical effects to Icariin, and from this point of view, the content of Epimedin C should be adopted as one of the quality indicating indexes. Meanwhile the essential oil in leaves of E.wushanense T.S.Ying was analyzed by GC/MS, and the result showed it consisted of acids,ketones,alkanes,alcohols,alkyne and other compounds.2. The value and importance of wine processed E.wushanense T.S.Ying was explored by applying modern scientific analysis technology. Firstly, the yellow rice wine made in Shaoshing was selected as best menstruum by integrating the two indexes which were icariin and total flavonoids set by Chinese Pharmacopeia; secondly, by referring to the ancient and modern literatures and integrating the relevant modern processing specifications, several wine processed products of E.wushanense T.S.Ying and E.brevicornu Maxim. were produced with the yellow rice wine, such as: wine-boiled product,wine-soaked product,wine-moistened product,wine- steamed product,wine-stir-fried product,wine-mutton -oil processed product. At the same time, the simple stir-fried product and the mutton oil processed product were used as the comparison group. Then, scientific indexes, namely Epimedin C,Icariin,total flavorins and polysaccharide, were chosen by combination with relevant pharmacological results. After content determination of the selected indexes, comparison was made between the decoction pieces of crude epimedium and that of the processed epimedium, furthermore the dissolution cases was also under investigation between them. Through Multifactorial Analysis, conclusion has been made: although variation law of different species of Epimedium differed to each other, wine processed Epimedium (including wine- steamed product,wine-stir-fried product,wine-mutton -oil processed product ) had displayed many advantages over the rest, even better than the mutton -oil processed product which is broadly being use in clinic. All the results showed that wine processed Epimedium was reasonable and of great value for medical usage, which accounted for its popularity in past dynasties, so it remained to be discussed whether this process method should be discarded or not. The primarily selected wine processed products need deeper study till the best one is determined which will raise the medical valuation of Epimedium and add one more processed product to the clinical practice. 3. the fingerprints of the processed epimedium was primarily explored by following steps: firstly, the fingerprints of flavonoids in E.wushanense T.S.Ying,crude epimedium and several varieties of other processed products was primarily established through HPLC gradient methods. Then, most of the common peaks, such as:Epimedin A,Epimedin B,Epimedin C,Icariin,Ikarisoside F,SagittatosideB and IcariinⅡ, in the fingerprints was recognized which accounts for nearly more than 75% of the gross peak area. By comparison: the fingerprints of E.wushanense T.S.Ying and E.brevicornu Maxim. had lots of peaks in common, but there were big differences between the peak area of both the peaks in common and peaks that were not in common, which indicated that the content of the flavonoids in different species differed greatly and corresponding HPLC fingerprints should be established respectively. In addition, the similarity of the fingerprints of 3 different batches of same decoction pieces from the same habitat which were processed by identical method was above 99%. And the result showed that decoction pieces of epimedium from the same habitat had stable quality, and the processing technology had excellent stability and reproducibility. Meanwhile, by comparing to the decoction pieces of the crude herbs, the peak area variation law of the processec products of E.wushanense T.S.Ying and E.brevicornu Maxim.was not completely the same, which also varied between the two species processed by the same way. It is also expected that further study should be made to illuminate variation mechanism of the chemical compositions in epimedium during the processing .we founded that there was obvious difference in the content and components-rate of change- discipline after precessed, and so was in water. So we advised that it shoud be established corresponding quality standard for several-origins Chinese Traditio and Medicine, in Chinese Pharmacopoeia (2005 edition) ,to ensure the quality stable and controllable, more to progress the clinical effect of Chinese Tradition and Medicine.