| Objective: We carried out a systematic study on the chemical constituents of Cymbidium Rhizoma,which will provide the data support for the study on the basis of the active substance,the quality evaluation and the further development and utilization of Cymbidium Rhizoma.On the basis of the study on the chemical constituents of Cymbidium Rhizoma,the quality standard of Cymbidium Rhizoma was established to provide a comprehensive method for evaluating the quality of Cymbidium Rhizoma.Methods:Cymbidium Rhizoma were extracted with 95% ethanol and then extracted into the concentrate.The concentrated was mixed with water to form a suspension,which is sequentially extracted with petroleum ether,ethyl acetate and n-butanol,Recovery reagent,each extraction part will be obtained.The extractionpartsof ethyl acetate and petroleum was separated and purified by variety of chromatographic techniques.The structures were identified by various methods that including thin layer chromatography,spectrum,mass spectrometry,literature comparison and so on.The volatile oil from Cymbidium Rhizoma was extracted by steam distillation and separated by GC-MS.The obtained mass spectra were analyzed by artificial analysis and NIST library.The retention index was used for qualitative analysis,and the relative content of each component was calculated by peak area normalization method.In accordance with the provisions of the< Chinese Pharmacopoeia >2015version,Cymbidium Rhizoma was identified by character,Microscopic and TLC;Theseparation was performed by HPLC gradient elution with the mobile phase composed of Acetonitrile(A)-0.04% trifluoroacetic acid water(B).The wavelength of detector was set at 264 nm.The mobile phase flow rate of 1 ml·min,columntemperature 30 ℃.The reference is 3,7-dihydroxy-2,4-dimethoxyphenanthrene.The HPLC chromatographic fingerprint of 10 samples of total phenanthrene position of Cymbidium Rhizoma were evaluated.Results: 17 compounds were isolated from the petroleum ether and ethyl acetate sites of Cymbidium Rhizoma,and 14 compounds of them were identified by modern spectroscopy and spectroscopic techniques.5 compounds isolated from petroleum ether,including two esters compounds:β-sitosterol palmitate(1),butyl phthalate(2);three steroids compounds:Stigmast-4-ene-3β,6β-diol(3),β-sitosterol(4),daucosterol(5).(6)9 compounds isolated from ethyl acetate,six phenanthrene compounds:4,5-dihydr-oxy-2-methoxy-9,10-dihydropheninthrene(6),Monbarbatain A(7),Ephemeranthoquino ne B(8),3,7-dihydroxy-2,4-dimethoxy-phenanthrene(9),Coelonin(10),Cymbinodin A(14);one amide compounds: N-trans-ferulic acid caramamide(11),small molecule phenolic acid compounds 2: p-hydroxybenzoic acid(12),trimesic acid(13).49 chromatogram peaks were isolated from the volatile oil of Cymbidium Rhizoma,and 33 compounds were identified,which accounted for about 94.248% of the total peak area.The results showed that the mainly volatile chemical constituents were fatty acids,esters,alcohols,aldehydes and alkanes class and so on.There were 8compounds with relative content more than 2%,namely Nerolidol(4.956%),Myristic acid(8.302%),Pentadecanoic acid(15.298%),Dissolute phthalate(8.850%),Palmitic acid(33.805%),Linoleic acid(4.030%),trans-13-octadecenoic acid(2.642%),Erucylamide(2.230%).The total ash content of Cymbidium Rhizoma should not exceed 10.64%,the acid insoluble ash content should not exceed 5.12%,and the content of the water-soluble extract(hot leaching method)should be less than 30.0%.A method of HPLC fingerprint of the total phenanthrene position of Cymbidium Rhizoma was established.By studying comparatively the fingerprints of ten samples,13 peaks have been identified as the general peaks,and analyzed the similarity of 10 samples’ fingerprint.The results of the analysis meet the requirements.Conclusion: Through this study,the 14 compounds isolated are the first to be isolated from Cymbidium faberi.The quality standard draft was established,which could be used to evaluate the quality of Cymbidium faberi. |
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