| Dendrobium officinale Kimura et Migo are commonly used herbal medicine,rare medicinal plants.And have been recorded Individually by the“Chinese Pharmacopoeia" in 2010 edition.are widely used as an Yin tonic to nourish the stomach and promote the production of body fluid.In this paper,leaves of Dendrobium officinale species in Yunnan as the chemical composition of the extraction source,Separate the chemical composition of the leaves from Dendrobium officinale,Then applied to the determination of the chemical composition of Dendrobium officinale stems from different producing area,and research the Pharmacological Pharmacodynamics effects.To provide a certain significance for the Dendrobium officinale medicinal herbs material basis of research,quality control and further development and utilization of resources.Objective:(1)To separate and purify flavonoid glycoside chemical composition from Yunnan Dendrobium provenance leaves.In order to obtain index components that can be used for quality control of Dendrobium.(2)To establish the method for the determination of flavonoid glycosides chemical composition from Stems of different provenance Dendrobium officinale.(3)To study pharmacological activity in vitro antioxidant of Yunnan provenance Dendrobium flavonoid glycosides.Methods:(1)we separated the leaves of Yunnan provenance Dendrobium by using 70%ethanol to heated at reflux for extraction,Water-saturated butanol to extraction,macroporous resin Chromatogram,Sephadex LH-20 Chromatogram,ODS Chromatogram,preparative liquid chromatography,RP-HPLC a variety of methods.Studied the structure by detecting Compound physicochemical properties,using modern means of various spectroscopic(UV、ESI-MS、1H-NMR and 13C-NMR).(2)Taking the isolated 4 flavonoid glycosides chemical composition from the leaves of Dendrobium officinale species in Yunnan as reference and established a method for the determination of Dendrobium flavonoid glycosides.Dendrobium stems and medicinal content of different provenances were measured and methodological study.(3)Experimental study on anti-oxidation effect of vitexin-2″-O-rhamnoside isolated from Yunnan kinds of flavonoid glycosides Dendrobium leaves.We measured Successively scavenging of Vitexin-2″-O-rhamnosides on DPPH·,ABTS ·,copper reducing power,and the effects of Hydroxy-induced on bone marrow mesenchymal stem cells viability oxidation damage.SPSS19.0 statistical software used to calculate half clearance and significant differences.Results:(1)Four flavonoid glycosides Compounds had been isolated from the leaves of Yunnan provenance Dendrobium officinale,By structural identificating,named:texin-4″-glucoside,vitexin-2″-O-xylopyranose,vitexin-2″-O-rhamnoside and isovitexin-2″-O-rhamnoside.(2)After analysis of chromatographic conditions,The separation of Vitexin-4″-O-glucoside and vitexin-2″-O-xylosidic could reach HPLC separation requirements for determination.the linear ranges of 8.75-280 μ g/mL(r=0.9999)、6.25-200 μ g/mL(r=0.9999)had a fine linear relationship.The average recoveries(n=6)are 100.62%、99.95%.RSD are 2.69%、2.06%.Vitexin-2″-O-rhamnoside and iso vitexin-2″-O-rhamnoside are isomers,the resolution failed to reach the baseline separation requirements,so we used mixtures(the concentration ratio is 1:1)as reference.the linear ranges of 12.5-400 μ g/mL(r=0.9999)has a fine linear relationship,the average recoveries(n=6)are 99.82%,RSD is 1.99%.Yunnan and Guangxi provenance Dendrobium officinale flavonoid Feature Components are similar,which all Contain vitexin-2″-O-rhamnoside and isovitexin-2″-O-rhamnoside,Yunnan Provenance Dendrobium officinale have higher Content.Zhejiang and Danxia Provenance Dendrobium officinale do not contain the chemical composition of vitexin-2″-O-rhamnoside and isovitexin-2″-O-rhamnoside or not be detected.(3)Vitexin-2 ″-O-rhamnosides had ertain clearly effect on DPPH radical,ABTS radical.When the concentration is 1 mg/mL,vitexin-2″-O-rhamnoside for DPPH radical scavenging rate is 56%,EC50 is 0.906mg/mL.When the concentration is 0.5 mg/mL,vitexin-2″-O-rhamnoside for ABTS radical scavenging rate is 52%,EC50 is 0.507mg/mL.We stablished Hydroxy-induced bone marrow mesenchymal stem cells oxidative damage model.Compared with the model group,the cell viability of administration concentration 50μ g/mL group is significantly increased(P<0.05),the cell viability of administration concentration 100μg/mL group is significantly increased(P<0.01)Conclusion:(1)Yunnan provenance Dendrobium officinale Leaves contain higher flavonoid glycoside chemical composition.The established method of extraction and separation of chemical composition is steady,This method can be used as a source of flavonoids of Dendrobium officinale medicinal ingredients characteristic reference substance.(2)Dendrobium officinale stems all contain four flavonoid glycoside chemical composition extracted from the Dendrobium officinale leaves,the separation of vitexin-4″-O-glucoside and vitexin-2″-O-xylosidase HPLC determination method is feasible,the determination of Vitexin-2″-O-rhamnoside and isovitexin-2″-O-rhamnoside need further study.Flavonoid content of Dendrobium officinale from different producing area and provenance is quite different.Which can be used as one of the bases for provenance judgment and quality control.(3)Vitexin-2″-O-rhamnoside have some antioxidant activity,Which provide a certain amount of research foundation for Dendrobium officinale pharmacological activities of other related flavonoids,and provide a certain amount of foundation for further development and utilization of Yunnan provenance Dendrobium officinale leaves. |
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