The Studies On The Chemical Constituents In Leaves Of Sorbus Tianschanica Ruper

Objective：1. Through the extraction, separation, structure identification of the chemicalcomponents of Sorbus tianschanica Rupr.2. The HPLC/MS and MEKC method for thedetermination which was the effective constituent of Sorbus tianschanica Rupr. wasestablished. Methods：1. Combined with the preliminary experiment of chemicalconstituents system, defined the chemical compositions of Sorbus tianschanica Rupr.Initially. Coarse powder of leaves and branches of Sorbus tianschanica Rupr. wasextracted by95%ethanol, and extracts were extracted by petroleum ether, ethyl acetate,n-Buthyl alcohol sequentially, then separated and isolated each part of extraction withsilica column chromatography method,on-step column chromatography, SephadexLH-20,thin layer chromatography method and recrystallization method to obtain monomericcompound. Using modern spectroscopic techniques (1H-NMR,13C-NMR), infraredspectroscopy and the combination of physical and chemical methods of structural analysisand identification to the monomeric compound.2. The contents of amygdalin and hyperinin branches and leaves, fruits of Sorbus tianshanica Rupr. were determined by LC/MSmethod with Hypersil BDS C18(150mm×4.6mm,5μm) column and mobile phase ofacetonitrile-0.1%formic acid (15:85), flow rate of0.8mL·min-1, column temperature of25℃. The conditions of mass spectrometry: the analytical technique of ions: selected ionmonitoring mode (SIM); ionic polar: negative ion detection mode; ionization method:electrospray ionization(ESI).3. Adopting micellar capillary electrophoresis method for thedetermination of the amygdalin content in the fruit of Sorbus tianschanica Rupr., andbuffer concentration, pH value, voltage and other influencing factors were analyzed toselect the best test method to draw the standard curve so that we can make a study onmethodology in analysis method. Results:1. Five compounds were isolated from thefoliage of Sorbus tianschanica Rupr., which through the identification shows itsstructure as n-octacosanol (Ⅱ) kaempferol-3-O-β-D-glucoside（Ⅲ）, hyperin（Ⅳ）, rutin(Ⅴ).2. The contents of hyperin in branches and leaves of Sorbus tianshanica Rupr.were higher than that in fruits, instead, the contents of amygdalin in fruits of Sorbus tianshanica Rupr. were higher than that in branches and leaves. And the contents ofamygdalin and hyperin in branches and leaves of Sorbus tianshanica Rupr. wererespectively0.452mg·g-1and1.236mg·g-1; the contents of amygdalin and hyperin in fruitsof Sorbus tianshanica Rupr. were0.727mg·g-1and0.36mg·g-1,respectively.3. UsingMEKC method to detect the amygdalin content is1.75mg·g-1of the Sorbus tianschanicaRupr. Fruit, detection wavelength was215nm, in the25~225μg·ml-1linear range of thesamples, and the average recovery was100.8%, RSD value was2.8%. Conclusion：1.Using a variety of natural medicine, chemical extraction, separation and purificationmethods, and application of modern spectroscopic techniques for the structuralidentification of the isolated compounds. kaempferol-3-O-β-D-glucoside（Ⅲ）, hyperin（Ⅳ）, rutin (Ⅴ) is the flavonoids. And n-octacosanol kaempferol-3-O-β-D-glucoside,rutin for the first time it is isolated from the foliage of Sorbus tianschanica Rupr., which toa certain extent gets a more complete study of the flavonoids of Sorbus tianschanica Rupr.,a more explicit understanding of the pharmacological activity, and sure it will promotefurther development and utilization of herbal medicines Sorbus tianschanica Rupr.2.The method is rapid and accurate, it may provide a new reliable means for the qualitycontrol of Sorbus tianschanica Rupr. preparations.3. The content of amygdalin in Sorbustianschanica Rupr. was determined by MEKC. This method is simple，rapid，lowerconsumption and the result is accurately reliable，which can be used for the determinationof cyanogenic glycosidesthe and the quality control of Sorbus tianschanica Rupr.