The Biological Activity Research Of Taishan Rhododendron Micranthum Turcz And The High-Speed Counter-Current Chromat Ography Separation Of Effective Ingredients

Objective:This research project is part of the work of "contains a side of Taishan Rhododendron micranthum Turcz composition of biological pesticide development"(No.2014 d056)and "the Insecticidal component separation and purification of Taishan Rhododendron micranthum Turcz"(No.2015 d065),Taian City college students science and technology innovation projects.It shall determinate the biological activities of Taishan Rhododendron micranthum Turcz branches and leaves on Musca domestica,and extract Taishan Rhododendron micranthum Turcz and determine its main active parts.It shall extract the main active part in Taishan Rhododendron micranthum Turcz by high-speed counter-current chromatography and get monomer compounds and confirmed its structure and determinate the biological activities of the monomer compounds.Methods:Through feeding the Musca domestica with the leaves and branches' powder of Taishan Rhododendron micranthum Turcz,it determinates the biological activity of Taishan Rhododendron micranthum Turcz on Muscadomestica.Extract the leaves and branches' powder of Taishan Rhododendron micranthum Turcz by petroleum ether Soxhlet extraction,and then soaking extract in 95% ethanol.Dry by distillation and use methylene chloride,ethyl acetate and n-butyl alcohol to flush the extractive with silica gel column.Then use the petroleum ether,dichloromethane,ethyl acetate and n-butanol extract parts to conduct bioactive experiment on Musca Domestica separately to determine the main active site.Screen out the high-speed counter-current chromatography condition of the active site and extract by the high-speed counter-current chromatography extraction.Then determine the biological activity of the extraction.Results:The minimum effective concentration of the leaves and branches' powder of Rhododendron micranthum Turcz inhibited obviously the oviposition of Musca domestica is 4%,valid for up to 60 h.The concentration of 5% the leaves and branches' powder of Rhododendron micranthum Turcz can cause a sluggish development and aprolonged larval period of Musca domestica,making its development period lengthen 0.8 d.The leaves and branches' powder of Rhododendron micranthum Turcz had no obviously effects on the Musca domestica pupation rate.Through the activity of Musca Domestica experiment,it determines the ethyl acetate extract parts is the mainly active site on Musca Domestica larvae.By optimizing the experimental comparison,we determined the high-speed counter-current chromatography extraction system of the ethyl acetate extracted parts of Taishan Rhododendron micranthum Turcz is that petroleum ether: ethyl acetate: methanol: water=5:5:4:6,stationary phase retention value is 55%,flow rate is 2.0 mL/min,the temperature is 25? and the rotate speed is 800 r/min.Through high-speed counter-current chromatography in ethyl acetate parts of the Taishan Rhododendron micranthum Turcz,we finally get a single compound,identified as quercetin.According to Musca Domestica larvae growth inhibition experiment of the two components of high-speed counter-current chromatography separation,its biological activity has not been proved.Conclusions:The branches and leaves of Taishan Rhododendron micranthum Turcz have good biological activity on Musca Domestica.High-speed counter-current chromatography with the suitable solvent system and experimental condition could separate the components of Taishan Rhododendron micranthum Turcz effectively.Due to non-solid supporter,high-speed counter-current chromatography could avoid sample loss,inactivation and degeneration caused by irreversible adsorption,therefore,it is especially suitable for the isolation of bioactive compounds from natural products.Significance:This research project first verifies the biological activity of Taishan Rhododendron micranthum Turczon on Musca Domestica,which is simple to operate and easy to develop.This research project first use high-speed countercurrent chromatography to extract Taishan Rhododendron micranthum Turcz and get a single compound laying a solid foundation for further development and utilization of Taishan Rhododendron micranthum Turcz.