Comparative Study Of Huangcao Shihu(Dendrobium Nobile,Dendrobium Fimbriatum,Dendrobium Denneanum)Based On The Fingerprinting Technology

Dendrobium generally be classified into two categories:dry goods and fresh goods,due to different species and processing method.Dry goods can be divided into Huancao,Huangcao,earring dendrobium(Fengdou),etc.From the earliest records,the origin of Huangcao Shihu was Dendrobium nobile;Dendrobium nobile was not the only Huangcao Shihu in 1930,Huangcao Shihu also includeds Mabiancao(Dendrobium fimbriatum in Guangxi and Dendrobium chrysanthum in Guangdong);in the 80's and 90's,it was reported that the origin of dozens Huangcao Shihu was not only including the previously reported and including Dendrobium denneanum and so on.This study used the technology of HPLC characteristic,thin-layer chromatography,UV spectrum to compare this three different kinds of Dendrobium on the basis of the early stage of the research,in order to achieve the purpose of identification.Objective1.To establish HPLC characteristic spectrum of Dendrobium nobile and study quality of Dendrobium nobile from different regions,to discuss its rationality provenance classification.From the differences in HPLC characteristic spectrum of fresh and dried Dendrobium nobile,To discuss the differences of chemical components from fresh and dried Dendrobium nobile;To confirm the chemical components of peaks from Dendrobium nobile;To compare HPLC characteristic spectrum of Dendrobium nobile,Dendrobium fimbriatum,Dendrobium denneanum by using the same chromatographic condition of HPLC characteristic spectrum of Dendrobium nobile.2.To establish HPLC characteristic spectrum of Dendrobium fimbriatum.3.To establish HPLC characteristic spectrum of Dendrobium denneanum.4.To compare difference TLC of alkaloids and other components among the three species.5.To compare difference UV Spectroscopy among the three species.Methods1.By using the HPLC characteristic spectrum technology to establish the analysis method of characteristic spectrum of Dendrobium nobile,Dendrobium fimbriatum,Dendrobium denneanum.The Similarity analysis by using the State Pharmacopoeia Commission Specific chromatogram similarity evaluation system software(2004A version).2.By using technology of HPLC-UV-ESI-MS to confirm one flavonoid chemical components of peaks from Dendrobium nobile.3.By using TLC of alkaloids and other components to identify the three species of Dendrobium by comparing the TLC spots location,colour,size and number,as the basis of identification.4.By using technology of UV Spectroscopy to measure the three Dendrobium ultraviolet spectrum and maximum absorption wavelength with different solvent(water,absolute ethanol,chloroform),as the basis of identification.Results1.Quality of Dendrobium nobile was much different from different regions,provenance had its regularity.(1)Integrate the HPLC Characteristic Spectrum of Dendrobium nobile from Chishui Guizhou,Yunnan,Guangxi,Hejiang Sichuan,provenance had its regularity.The HPLC Characteristic Spectrum of Dendrobium nobile could be divided into two major classes,one was from Chishui Guizhou Province,one was from Yunnan Province,samples from Hejiang Sichuan similared to Chishui Guizhou Province,samples from Guangxi included both.10 batches of Dendrobium nobile from Chishui Guizhou were marked with 22 common specific chromatographic peaks,and the similarities were 0.979?0.998.10 batches of Dendrobium nobile from Yunnan were marked with 27 common specific chromatographic peaks,and the similarities were 0.902?0.980.(2)There were little difference between fresh and dried Dendrobium nobile of the main characteristic peaks and relative peak area,the similarity reached 0.989.(3)A flavonoid of apigenin-6,8-?-C-? D glucopyranoside from Dendrobium nobile was initially determined.Samples from each growing area all had this specific chromatographic peaks.(4)There were great differences among the three species of chemical composition,however the three species contained dendrophenol.2.HPLC Characteristic Spectrum of Dendrobium fimbriatum were established.HPLC characteristic spectrum of 10 batchs of Dendrobium fimbriatum were marked with 9 common specific chromatographic peaks,and the similarities of of samples were 0.837?0.992.And preliminary study determined it not detecting characteristic peak of flavonoids.3.HPLC Characteristic Spectrum of Dendrobium denneanum were established.HPLC characteristic spectrum of 10 batches of Dendrobium denneanum were marked with 12 common specific chromatographic peaks,and the similarities of 10 batchs of samples were 0.846?0.986.It clearly identified dendrophenol and detected one characteristic peak of flavonoids,there was one characteristic peak that the single-peak area accounts for 61 percent of common peak area.4.There were great differences of TLC among Dendrobium nobile,Dendrobium fimbriatum,Dendrobium denneanum.(1)Dendrobium nobile contained dendrobine TLC spot,Dendrobium fimbriatum,Dendrobium denneanum didn't contain it.(2)Dendrobium denneanum contained dendrophenol TLC spot,Dendrobium nobile,Dendrobium fimbriatum had the different colour spot in the location of dendrophenol,it may be the interference of other chemical composition.5 There were some difference of UV Spectroscopy among Dendrobium nobile,Dendrobium fimbriatum,Dendrobium denneanum.Viewing the UV Spectroscopy spectrogram of water system,absolute ethanol system,chloroform system separately,the UV Spectroscopy spectrogram of the three species showed differene peak include in number,shape and intensity.Conclusion1.The method of HPLC characteristic spectrum of Dendrobium nobile,Dendrobiumfimbriatum,Dendrobium denneanum can reflects categories and relative quantities of mainchemical composition of the three species of dendrobium,it can be used as the reference for identification.2.Using TLC and UV Spectroscopy,it can be used as the reference for identification.