| Objective:Sophora Flavescens Ait mainly contains two types of chemical components:Alkaloid and Flavonoids ingredients. At present, there are many researches for Sophora Flavescens Ait at home and aboard. However, most of researches concentrate at Alkaloid Components. There are less researches on Flavonoids ingredients of Sophora Flavescens Ait.The previous study confirmed that Matrine and Kurarinol extracts have a role in calming and sedative effects.Alkaloids like Matrine and Kurarinol, are produced by separating tranquilizing effective parts of Sophora Flavescens Ait. Flavonoids ingredients and other types of components are produced by the same method. At present,tranquilizing effect of Flavonoids ingredients Components in Sophora Flavescens Ait has not been reported. This experiment is studying on Flavonoids ingredients Components in Sophora Flavescens Ait. Extracting and separating of Flavonoids ingredients Components from tranquilizing effective components in Sophora Flavescens Ait. The aim is to study its effect on the central nervous system and further elaborate tranquilizing effects of Sophora Flavescens Ait.Methods:This experiment extracts and separates Sophora Flavescens Ait by using systematical solvent separation method. Preliminary screening tranquilizing effective parts in Sophora Flavescens Ait by using righting reflex test. Comparative observation of developing solvent and color reagent in Matrine, Oxymatrine and Flavonoids ingredients. Detection conditions of thin-layer chromatography of Sophora Flavescens Ait are established, and detecting Matrine, Oxymatrin and Flavonoids ingredients in dichloromethane tranquilizing effective parts. The HPLC detection conditions of Flavonoids ingredients are explored. Methanol-Water, Acetonitrile-0.2% Formic Acid are investigated separately on the choice of mobile phase. Wave length, such as 280nm,320nm,30nm, is detected and screened, and Flavonoids ingredients chromatogram of extraction parts in Sophora Flavescens Ait is established. Based on the high efficient countercurrent chromatography, two items of solvents are investigated separately on system, velocity, speed and experimental temperature, improving chromatographic conditions,establishing separation methods what are easy to control, stable and repeatable, are used in Sophora Flavescens in Sophora Flavescens Ait extracts,and preparation of Sophora Flavescens; Cell experiment is used to study the effect of hippocampal neurons of Brass Components in Sophora Flavescens Ait.Results:Through the system solvent for extraction of Sophora Flavescens Ait get a sediment (Sophora Flavescens Ait no.1), Sophora Flavescens Ait water extraction liquid dichloromethane extract (Sophora Flavescens Ait no.2), Sophora Flavescens Ait water extraction liquid ethyl acetate extract (Sophora Flavescens Ait 3), Sophora Flavescens Ait water extract(Sophora Flavescens Ait no.4), Sophora Flavescens Ait alcohol extraction liquid dichloromethane extract (Sophora Flavescens Ait no.5), Sophora Flavescens Ait alcohol extract ethyl acetate extract (Sophora Flavescens Ait no.6), Sophora Flavescens Ait alcohol extract (Sophora Flavescens Ait no.7). By the vivo animal experiment screening from Sophora Flavescens Ait extract tranquilize mind effective parts (P< 0.01), Is the alcohol extraction dichloromethane extract part (Sophora Flavescens Ait no.5). Established the matrine, Oxymatrin thin layer chromatography detection conditions,the developing solvent is a mixed system of dichloromethane-methanol-strong aqua (5:0.6:0.3), the color reagent for bismuth potassium iodide try fluid and sodium nitrite ethanol solution, Can show lasting orange spots. Established the flavonoids ingredients of thin layer chromatography detection conditions, the developing solvent is a mixed system of petroleum ether-ethyl acetate-methanol solution, chromogenic agent for aluminium trichloride ethanol solution, Set can be observed under uv light (365 nm) clear spots, And detected dichloromethane parts tranquilize mind active site of matrine, Oxymatrine and flavonoids ingredients. Established the sophora flavonoids ingredients high performance liquid chromatography (HPLC) method, By investigating the established the acetonitrile-0.2% formic acid water as mobile phase,360 nm for sophora flavonoids ingredients detection wavelength,and Flavonoids ingredients chromatogram of extraction parts in Sophora Flavescens Ait is established. Through to the flavonoids ingredients research of high-speed countercurrent chromatography conditions, determine the mobile phase for the petroleum ether:ethyl acetate:methanol:water (1:1:1),. Speed:800 r/min, Velocity:3ml/min, Temperature 25℃, The detection wavelength of 365 nm. By high-speed counter-current chromatography to get two monomer component and multiple components.,The flavonoids sophora identified by thin layer chromatography (TLC) to detect flavonoids component (no.1-5). Through the cell experiment results confirm sophora flavonoids component 3 compared with the blank group has a very significant difference (P< 0.01). Experiments prove that sophora flavonoids ingredients have the effect that are activated to restrain activity of nerve cells in the hippocampus.Conclusion:Through vivo animal, identifying that dichloromethane parts in Kurarinol are tranquilizing effective parts. Monomer components and several Flavone components of separating tranquilizing effective parts have been gotten by using high-speed countercurrent chromatographic conditions. Flavonoids ingredients have effect of inhibiting hippocampal neurons cell activity. Meanwhile, methods such as thin-layer chromatography, high performance liquid chromatography, which can be used to determinate matrine, Oxymatrine and Flavonoids ingredients in active parts are established. |
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