Study On The Separation And Purification Of Resveratrol From Polygonum Cuspidatum Sieb. Et Zucc.

Based on a large number of literature,resveratrol(Res),with important pharmacological action of anti-cancer and anti-heart brain blood vessel disease, was selected in this dissertation to be the research object ,and Polygonum cuspidatum Sieb. et Zucc. as the raw material.Deep and systematical researches had done to the extraction > isolation and purification of Res from Polygonum cuspidatum Sieb. et Zucc. Process route, condition, analytical method, isolation and purification factors were studied and established. To develop new isolation method for natural products with high efficiency,the application of molecularly imprinted technique(MIT) to the separation of effective components in natural products was also studied. Details are as follows:1.Efficient analytical methods of effective components in Polygonum cuspidatum Sieb. et Zucc.were developed in a general and system way. Simultaneous and quick determination of Res and Polydatin (Pol) in Polygonum cuspidatum Sieb. et Zucc. by HPLC was presented;HPLC analysis of Emodine(Emo) in Polygonum cuspidatum Sieb. et Zucc. and analysis of effective components in Polygonum cuspidatum Sieb. et Zucc. by TLC were also established;The stability of Res,in the condition of different pH ,temperature and light radiation were discussed.2.Study on the extraction techniques.The extraction condition of effective componets from Polygonum cuspidatum Sieb. et Zucc. was investigated by using the orthogonal desigh with Pol,Res and Emo as index; In according to the structure of effective component,using a complex enzyme,a new technology of extracting Res from Polygonum cuspidatum Sieb. et Zucc.was developed; The solvent extraction technique of Res and Emo after enzymatic treatment was optimized , which made both the extraction ratio and quality of products improved greatly; Kinetics of enzymatic hydrolysis was discussed.The results showed that enzyme-based extraction technology was characterized by its high extracting ratio and gental extracting conditions determinedexperimently.Compared with traditional extracting technology by ethanol,the extracting ratio of Res was increased from 2.40mg/g to 12.13mg/g, about 5 times by using this enzyme-based extracting technology. Through optimazation of extraction process after enzymatic treatment ,the yield and quality of Res were both improved, which have some reference value for enzymatic extraction of other traditional Chinese medicine.3.Study on the isolation and purification of Res and Emo in ethanol extracts after enzymatic treatment.Firstly,through pre-treatment, Emo,including most of other impurities were isolated from the extracts. Trough silica chromatography ,Emo, the by-product, was obtained with the content of up to 95% above.Secondly,at the first time,purification of Res with macroporous adsorptive resin was studied systematically. 1# resin as enrichment column and 2# resin as purification column in series was adopted.Res was adsorbed on 1# column at first with the obviation of impurity in large amounts.Then,impurity in small amounts was adsorbed on 2# column which adsorb much impurity, but little Res,so that Res was purified more and the content of Res was increased from 40.2% to 94.5%.Res with high purity of more than 99% can be obtained with further purification.5.The thermodynamic and kinetics behavior of Res on 1# resin was investigated by static method and kinetics experiment. Through the test of AQ AH, AS and the foundation of mathematical mode for transportion equation of Res,adsorption mechanism and the controlling step of transportion rate were discussed and identified,respectively.It gave a reasonable explanation on the adsorption behaviour of Res on 1# resin and was helpful for increasing adsorption and transport property.6. MIT was developed with an increasing speed recently .In order to expand the application of this technology in natural products,a molecular imprinted polymer(MIP) was prepared by a molecular imprinting technique using Res as the template molecular at the first time.The molecular imprinted polymer was investigated in equilibrium binding experiments to evaluate its molecular recognition, binding characteristics and solid phase extraction(SPE) property.The resultsshowed that,compared with non-imprinted polymer,the imprinted polymer exhibited much higher affinity for Res among other components, with a Res-poi=3.73, a Res.BPA=6.22,respectively .Application of the imprinted polymer to the separation of Res from complicated samp\es(Polygonum cuspidatum Sieb. et Zucc.extracts and peanut root extracts) showed the promising future of MIT in traditional Chinese medicine area.Anymore,through the methods of UV spectra,IR spectra and SEM,MIP was proved to be a multi-porous polymer ,which could be a explanation of its high affinity to template molecular through the functional groups and stero-structure.In this dissertation, with the combination of enzymatic technology and macroporous adsorptive resin technology,Res in Polygonum cuspidatum Sieb. et Zucc.was extracted and prepared. What's more,another effective compqnent,Emo, was also isolated , which made the raw material utilized efficiently.The application of MIP-SPE to the isolation of Res in complicated samples provided theory and practice base and also a new route for the separation and purification of effective components in natural products.