Study On Separation And Purification Of Chemical Constituents And Bioactivity Of Osteon Myospalacem Baileyi

In this thesis, Osteon Myospalacem Baileyi is chosen as object and studied from the following four aspects: selecting the extraction methods, building various modes of 2D liquid chromatography methods, preparation of pure compounds from the complex extract and preliminary screening of activity with preparative high performance liquid chromatography and two-dimensional liquid chromatography, and laid the material foundation for the development of new drugs.By studying the factors that affect the extraction of Osteon Myospalacem Baileyi, such as the extraction method, the extraction solvent, the extraction time, the extraction number and the liquid ratio, optimize the extraction process, to get the best extraction process. DAISO Silica normal phase column for one-dimensional separation and preparation of alcohol extract, DAISO C18 reversed phase column for water extract. Different elution procedure result in different separation effect, and ultimately determine the best preparation method. Normal phase preparative chromatography get 10 components, and reversed phase preparative chromatography get 5 components. Samples were initially divided, improves the efficiency of the preparation for the subsequent two-dimensional separation.Established a new type of 2D-NPLC / HILIC chromatography preparation system with DAISO Silica column and YMC Diol column, 2D-NPLC / NPLC chromatography preparation system with two Silica columns. It has good selectivity and orthogonality, achieved efficient separation and preparation of small and medium-polar substances of Osteon Myospalacem Baileyi alcohol extract component. Received a total of 16 high-purity compounds, some more than 98% purity. The structures were identified combined with mass spectrometry and NMR data, the results showed that the main components of alcohol extract were sterols and organic substances.Established 2D-RPLC / RPLC chromatographic system with high-density bonded polar hydrophilic MP C18 column for separating effectively complex components of water extracts. Preparative chromatography get 9 compounds with high purity, structure identification to get the sequence, the results showed that the main components of water extract were peptides, molecular weight is about 1kD,also providing new material basis for further study of pharmacological and toxicological activity of Osteon Myospalacem Baileyi.Experiment of preliminary screening activity that Osteon Myospalacem Baileyi extracts on the front osteoclasts(RAW264.7 cells), showed that six components of ethanol extract have significantly inhibited the proliferation of pre-osteoclast, which suggests that further research can be developed for osteoclast inhibitors.