Preliminary Study On Fingerprints And Chemical Constituents Of Dendrobium Medicinal Materials

Dendrobium is commonly used traditional Chinese medicine.There are 76 kinds of Dendrobiums in China,about 40 kinds of them are medicinal Dendrobiums.The medicinal history of Dendrobium is very long.China's first pharmaceutical monograph"Shen Nong's Materia Medica",there are Dendrobium " Remove the paralysis,lower adverse energy,make up the five dirty consumptive,thin and weak,nourishing kidney yin.Long-term consumption can be thick stomach,light body and longevity.The 2015 edition of "Chinese Pharmacopoeia" contains 4 varieties of Dendrobiums,Dendrobium nobile Lindl.,Dendrobium chrysotoxum Lindl.,Dendrobium fimbriatum Hook.and Dendrobium officinale Kimura et Migo,among which Dendrobium officinale is separately listed in the Pharmacopoeia.Dendrobium officinale,the dry stem of Orchidaceae plant Dendrobium officinale Kimura et Migo,which is one of the rare medicinal herbs in China with unique medicinal value.Dendrobium officinale contains polysaccharides,flavonoids,alkaloids,phenanthridines,bibenzyls,amino acids and other active ingredients.Modern research shows that Dendrobium officinale has anti-oxidation,anti-tumor,regulating immunity,relieving fatigue,treating diabetes and other pharmacological effects.Dendrobium officinale are mainly distributed in Anhui,Zhejiang,Hunan and Yunnan in China.Among them,Dendrobium Officinale Kimura et Migo in Huoshan is also called ‘wanzhangxu',which has been paid much attention by community as the authentic medicinal materials of Anhui Province,it is also regarded as the top grade of Dendrobium by ancient researchers.In this paper,Huoshan Dendrobium officinale,Dendrobium officinale from other areas and other varieties of Dendrobiums were compared in terms of HPLC fingerprinting,similarity evaluation and cluster analysis to provide reference for the quality identification,quality control and comprehensive utilization of resources of Dendrobium officinale.At the same time,the chemical constituents of four species of Dendrobium from Huoshan were analyzed by LC-MS and other methods.A method for the determination of total flavonoids,polysaccharides,naringenin and dendrobrin in different Dendrobium samples was established,and the contents of different Dendrobium samples was analyzed.On the different parts of Huoshan Dendrobium candidum immunomodulatory activity screening,screening of antioxidant activity,to clarify the Huoshan Dendrobium officinale immunomodulatory material basis forfurther study provide a reference.The main research contents are as follows:1,Fingerprints of Dendrobium officinale from different areas.The HPLC fingerprinting of methanol extract of Huoshan Dendrobium officinale from different areas was established,and a total of 12 common peaks were identified.The total peak area of ?? Huoshan Dendrobium officinale was higher.SPSS cluster analysis showed that Huoshan Dendrobium officinale samples could be divided into two types,5 batches of Huoshan Dendrobium officinale as a category,other areas of Dendrobium officinale for a class.Fingerprint similarity analysis showed that the similarity of Huoshan Dendrobium officinale to common mode was very high,ranging from 92.6% to 94.8%,indicating that there was no significant difference between Huoshan Dendrobium officinale.The similarities of Dendrobium officinale from Lin'an,Yueqing,Yancheng and Yunnan were between 17.5% and 41.5%,indicating that there were some differences in the HPLC fingerprints of Dendrobium officinale from different areas.Huoshan Dendrobium officinale had certain peculiarities compared with other Dendrobium officinale.Principal component analysis showed that Dendrobium officinale samples could be divided into two types: five batches of Huoshan Dendrobium officinale as one species and other Dendrobium officinale as other species.The principal component analysis results are consistent with the cluster analysis results.2,HPLC fingerprints and LC-MS analysis of methanol extracts of four kinds of Dendrobium from Huoshan.Taking the samples of Huoshan Dendrobium officinale as the reference,the HPLC fingerprints of different species of Dendrobium officinale methanol extract were studied and the similarity was studied.The results showed that the similarities between Huoshan Dendrobium officinale,Dendrobium moniliforme,Dendrobium huoshanense and Dendrobium henanense from Huoshan were higher.HPLC fingerprint of four species of Dendrobium chrysanthum from Huoshan was established.By LC-MS comparison,a total of 32 common peaks were identified.Thirteen common compounds were identified,which were 2,4,7-trihydroxyPhenanthroline,trans-N-(4-hydroxyphenethyl)ferulic acid amide,3,4'5-trihydroxy-3'-methoxybibenzyl,Dendrocanin C,Naringenin,Dihydrocortisone Dihydroxycinnamic Acid Ester,(R)-3,4-Dihydroxy-5,4 ',?-Trimethoxybibenzyl,Uridine,4' Dihydroxy-3,3'-dimethoxybibenzyl,3,4'-dihydroxy-5-methoxybibenzyl,3,4-dihydroxy-5,4'-dimethoxydi Benzyl,Denbinobin.The results of SPSS clusteranalysis showed that Dendrobium samples could be divided into two types,five batches of Huoshan Dendrobium officinale were the first type,the other dendrobium was the second type,the second type was Dendrobium moniliforme,Dendrobium huoshanense and Dendrobium henanense.Fingerprint similarity analysis showed that the similarity of Dendrobium moniliforme,Dendrobium huoshanense and Dendrobium henanense between 73.0%-80.7% in comparison with the common pattern of Huoshan Dendrobium officinale.It showed that the four kinds of Dendrobium from Huoshan have some similarities.Principal component analysis showed that Dendrobium officinale samples could be divided into four categories: Dendrobium moniliforme,Dendrobium huoshanense and Dendrobium henanense and Dendrobium officinale.The principal component analysis results are consistent with the cluster analysis results.3,Analysis of naringenin,gigantol,total flavonoids,polysaccharide content of dendrobium herbs.HPLC-UV method was used to determine the content of naringenin and gigantol in Dendrobium officinale from different areas,Dendrobium moniliforme,Dendrobium huoshanense and Dendrobium henanense from Huoshan,Dendrobium nobile,Dendrobium chrysotoxum,Dendrobium fimbriatum,Dendrobium devoninum,Huoshan Dendrobium officinale in the content of naringenin and gigantol more than other Dendrobium officinale.Gigantol was detected only in 5 batches of Huoshan Dendrobium officinale and Dendrobium moniliforme,Dendrobium huoshanense,Dendrobium henanense from Huoshan.Determination of total flavonoids in different Dendrobium by UV spectrophotometry.five batches of Huoshan Dendrobium officinale had higher total flavonoids content of 3.93-4.48 mg / g in the 12 batches of Dendrobium officinale,and the others was 0.79-2.13 mg / g.Determination of polysaccharides in different Dendrobium by UV spectrophotometry.In the 12 batches of Dendrobium officinale samples,the polysaccharide content of five batches of Huoshan Dendrobium officinale was very low,ranging from 150.37 to 239.41 mg / g and the polysaccharides content of Dendrobium officinale from other producing areas was 279.47 to 403.22 mg /g.The quality of Dendrobium officinale polysaccharides cultivated and produced in the planting base was generally higher than that of the wild herbs,and the breeding of Dendrobium officinale increased the polysaccharide content.The Huoshan Dendrobium officinale grown in wild conditions,so the polysaccharide content is low,the total flavonoids,naringenin,gigantol and other small molecule content higher.4,Study on antioxidation and proliferation activity of immune cells in vitro ofdifferent extraction parts of Huoshan Dendrobium officinale.DPPH free radical scavenging ability determination,· OH radical scavenging ability determination,reducing force performance determination and total antioxidant activity determination method were used to study the antioxidant activity of various parts of Huoshan Dendrobium officinale in vitro.N-butanol and ethyl acetate sites of Huoshan Dendrobium officinale have good DPPH scavenging capacity,second only to Vc,where ethyl acetate> n-butanol site.Huoshan Dendrobium officinale restoreability of various parts of the body: n-butanol> ethyl acetate> total water extract> residual water> crude polysaccharide.Total antioxidant capacity of Huoshan Dendrobium officinale:n-butanol> ethyl acetate> remaining water> total water extract> polysaccharide.· OH scavenging capacity of Huoshan Dendrobium officinale more obvious.MTT assay of various parts of Huoshan Dendrobium officinale promote the proliferation of ANA-1,the results showed that the water extract at a concentration of400 ?g / mL,can significantly promote the proliferation of ANA-1,when the concentration is too high,its growth will be inhibited;Crude polysaccharide at the concentration of 400 ?g / mL,600 ?g / mL,can significantly promote the proliferation of ANA-1;Huoshan Dendrobium officinale ethyl acetate in the role of the concentration of 25 ?g / mL,50 ?g / mL,100 ?g / mL can significantly promote the proliferation of ANA-1,when the concentration is too high,its growth will be inhibited;n-butanol concentration of each site can significantly promote the proliferation of ANA-1,of which 100 ?g / mL,promote proliferation The most significant effect;the remaining water site at the concentration of 100 ?g / mL,200 ?g / mL,400 ?g / mL,can significantly promote the proliferation of ANA-1,when the concentration is too high,its growth will be inhibited.