Study And Applicat Ion Of Three Dendrobum Species Of "Shuicao Fengdou" By Characteristic Spectrum Technology

Dendrobium,also known as“Shihu”in Chinese,is a commonly used and precious herb in Traditional Chinese Medicine.According to the record of China Pharmacopeia(2010 version),D.nobile,D.chrysotoxum,D.fimbriatum,and the related species of Dendrobium genus are all officially used as Shihu,and D.officinale is listed separately.Dendrobium is now rare in the wild.To meet the market demand,leading many other Dendrobium plants began to be substituted materials.Shihu Fengdou is one of the major products in the market and is very popular.Beside "Tiepi Fengdou",the principal commodities of these materials include:"Huodou","Zipi.Fengdou","Tongpi Fengdou","Shuicao Fengdou","Gangjie Fengdou" and "Chongcao Fengdou"."Shuicao Fengdou" is regarded as one of the representative type of commercial Fengdou,which are processed from D.crepidatum,D.aphyllum and D.chrysanthum.For their similar morphologies,it is so difficult to discriminate exactly from each other.Because of the diversity and similarity,the lack of effective quality evaluation criteria and regulatory,resulting in confusion and serious shoddy phenomenon in the market.Currently,few quality control studies were carried on this three Dendrobium species.This study used a combination of traditional Chinese medicine fingerprint technology and thin-layer chromatography method to explore the control system of the three Dendrobium species,in order to provide a useful reference for the Dendrobium quality control and standardization of the market,as well the depth of the utilization of products.Objective(1)To establish HPLC characteristic spectrum of D.crepi datum,D.aphyllum and D.chrysanthum.To explore their inherent characteristics.To compare the similarities and differences between them and to discuss its rationality and feasibility of the classification.(2)To establish HPLC characteristic spectrum of the“Shuicao Fengdou”that collected from Qingping market and some pharmacies,and to preliminary study the current market conditions of "Shuicao Fengdou".(3)To compare alkaloids and other components among the three species by using the thin-layer chromatography(TLC)method and provide a scientific basis for the TLC study of Dendrobium plants.Methods(1)Use the HPLC characteristic spectrum technology establish the analysis method to detect multiple batches of D.crepidatum,D.aphyllum and D.chrysanthum.Use the State Pharmacopoeia Commission Specific chromatogram similarity evaluation system software(2004A version)generate the Common pattern of HPLC characteristic spectrum,respectively,and calculate the similarity,compare the similarities and differences.(2)Establish HPLC characteristic spectrum of the "Shuicao Fengdou",use software of SPSS 20.0 for cluster analysis.Compare to the common pattern of the three Dendrobi um species,calculate the si mi larity,and determine their plant-based sources.(3)Use TLC of alkaloids and other components to identify different species of Dendrobiumby comparing the dot location,size and number,and compare with D.nobile of the alkaloids components which were officially recorded in the Chinese Pharmacopoeia.Results(1)HPLC characteristic spectrums of D.crepidatum,D.aphyllum and D.chrysanthum were established respectively.12 batches of D.crepidatum were marked with 41 common specific chromatographic peaks,and the similarities were 0.901?0.986.10 batches of D.aphyllum were marked with 33 common specific chromatographic peaks,and the similarities were 0.794?0.978.10 batches of D.chrysanthum were marked with 21 common specific chromatographic peaks,and the similarities were 0.939?0.989.The similarities among their three common patterns were very low,only 0.024?0.135.The result indicated that three species of Dendrobium have a high intraspecific similarity,but different Dendrobium species have different chemical components.(2)The HPLC characteristic spectrum of 24 batches of Shuicao Fengdou samples were classified into five clusters by cluster analysis.The cluster I(included 3 batchs)and the cluster ?(included 16 batchs)were broadly consistent with the D.aphyllum.The similarities with the common pattern of D.aphyllum were 0.657?0.880.The cluster?(including 1 batch)was basically consistent with the D.chrysanthum,the similarity was 0.906.10 batches were confirmed to establish the common pattern from the cluster?,33 specific chromatographic peaks were marked and the similarties are 0.970?0.998,and the similarity with the common pattern of D.aphyllum were 0.725.(3)There is an apparent difference in the TLC analysis,thus this methods can effectively identify the three Dendrobium species.TLC of alkaloids compared with D.nobile,only D.nobile has the same spot in the location of dendrobine.D.crepidatum and D.aphyllum have the same colour spot in the appropriate lacation of gigantol.Conclusion(1)In this paper,the feasibility and advantages of HPLC chromatographic spectrum were investigated and the technique was shown to useful for evaluation of D.crepidatum,D.aphyllum and D.chrysanthum.The method offers an alternative qualitative profile and can reflecting the characteristic and differences of various Dendrobium species.TLC was applied to the identification of the three Dendrobium species.This method is simple,sensitive and reliable,can be used to fast distinction.(2)The commercial "Shuicao Fengdou" were studied by HPLC characteristic spectrum.About 80%of Shuicao Fengdou may processing from D.aphyllum.The products market are complex,have large difference in quality and price.(3)This article provided an experimental basis and technical methods for the identification of D.crepi datum,D.aphyllum and D.chrysanthum and had important reference value.It is urgent to take concrete and effectibe measure to standardize the market,as to facilitate further development and ensure the clinical efficacy of Shihu.