Research On The Chemical Constituents And Biological Activity Of Echinops Ritro L.

The Echinops L.can be widely used for dispelling ecchymoma,anti-inflammatory,analgesic,and also for the treatment of a virety of diseases,such as migraine,tuberculosis,leprosy,schistosomiasis,syphilis,liver disease,stomach disease and so on.As a kind of folk medicine,the Echinops L.has a high medicinal value.However,the species of Echinops L are various,besides,misuse and mixed phenomenon is more serious in folk.Moreover,in quality control,the active ingredients of Echinops L are not clear and only using a few of chemical components as the standard of quality control can’t reflect the quality of herbs comprehensively.Domestic and foreign studies have shown that the chemical components of Echinops L.are extremely complex and have diverse pharmacological activities.Thus,how to effectively use modern extraction and separation technology system was carried out on extraction and separation of medicinal ingredients in the Echinops L.,simultaneously,how to reasonable development and utilization of medicinal resources of Echinops L.has become an important topic in the contemporary.At present,the study of Echinops L.mainly concentrated in the development and utilization of medicinal ingredients in Radix Echinopsis.The studies found that Radix Echinopsis contained thiophene,terpene,naphtha,and had antitumor,antiviral,antifungal,anti-inflammatory and other activities.However,the research on its chemical compositions and biological activities is rarely reported at home and abroad.Based on the research status of Echinops L.,in order to find the new active ingredient and clear the medicinal parts,the chemical composition and bioactivity of blue thorn in Xinjiang were studied systematically in our research.Moreover,fingerprint was systematically investigated to elucidate the important role of the whole herb and establish its scientific quality control method.The results are as follows:1.The air-dried aerial parts of the plants(200g)were hydro distillated for 4 h using a clevenger-type apparatus to produce essential oil according to the method.34 components were idectified by Gas chromatography–mass spectrometry(GC–MS)analysis,which represents 93.32% of the oils from Echinops ritro L..These analyses revealed that the most important feature of the oils was presence of a percentage of oxygenated sesquiterpene(approximately 22.88%),followed by the oxygenated monoterpenes and sesquiterpene hydrocarbons(9.86 and 11.49% respectively).The dominant components of Echinops ritro L.EO are a-pinocarvone,a-terpineol,nerolidol,and machilol.2.The seventy percent of ethanol extracts of the blue thorn in Xinjiang were chemically analyzed through a variety of chromatography methods.Finally,40 compounds were isolated from the whole herb.Based on the physical and chemical properties and spectral analysis,the structures of 31 compounds were determinedas follows,including 10 flavonoids: Quercetin(1),Hyperoside(2),QuerciMeritroside(3),Isorhamnetin(4),Luteolin(5),luteolin-7-O-β-Dglucoside(6),Apigenin(7),Apigenin-7-O-β-D-glucoside(8),kaempferol-3-O-galactoside(9),Chrysoriol(10);4 Steroids: Ergosterol(11),Daucosterol(12),beta-Sitosterol(13),Stigmast-5-ene-3β,7α-diol(14);3 triterpenoids: Ursolic Acid(15),missouriensin(16),Betulonicacid(17);12 phenol acids:Syringic acid(18),trans-p-Hydroxycinnamic acid(19),3,4-Dihydroxybenzoic acid(20),Vanillin(21),Ferulaldehyde(22),p-Hydroxybenzaldehyde(23),3,5-Dimethoxy-4-hydroxybenzaldehyde(24),Pyrogallol(25),Gallic acid(26),α-hydroxytetracosanoic acid(27),Decyl myristate(28)Dibutyl phthalate(29);two other kinds of compounds: Nonacosyl(30),Hentriacontan-2-one(31).Among the compounds,10 of which were isolated from the genus of Echinops L.for the first time,and 15 of which were isolated from the blue thorn in Xinjiang for the first time.3.The HPLC fingerprint analysis method was successfully established and applied to three batches of blue thorn in Xinjiang for the quality control.Using the chromatogram fingerprint of traditional Chinese medicine similarity evaluation system,three batches of samples were classified and evaluated.Compared with reference substances,10 compounds were identified.4.The EO exerts potent inhibitory effects against E.coli,S.aureus,and Salmonella enteritidis with MIC values of 2.5,0.15,and 0.6 mg/mL;also,the MBC values were 7.5,1.0,and 3.0 mg/mL.Results showed that Echinops ritro L.EO possesses remarkable antibacterial and bactericidal properties.Observations of changes to the cell microstructure,biofilm inhibition,cell viability assay,and whole-cell proteins methods provide a further means to study the functional mechanism.The results showed remarkable antibiofilm and disruption of the bacterial membrane.Therefore,Echinops ritro L.EO represents a source of natural compounds with the potential use in food systems to prevent the growth of foodborne bacteria and to extend the shelf life of processed food.5.We also analyzed their anti-tumor activity of two human tumor cell lines HepG2 and HeLa via MTT assay.Compound 2,4,6,8,9 and 10 presented effective cytotoxicity against human tumor cell lines HepG2 and HeLa.