Study The Extraction、Purifition、Preliminary Identification Of Flavonoids From Amoroha Fruticosa L.Leaves

Amorpha fruticosa L. is Leguminous plants,have been grown in the north and the south. In the highway landscaping, has been widely used in landscape gardening. It has a strong solid soil slope protection, improved the ability of the soil. Flavonoids are plant secondary metabolites, widely exists in green plants, Especially in the Compositae,Leguminosae, Rutaceae, Scrophulariaceae and Labiatae content is higher than other plants. Flavonoids in the process of rhizobium infect with instructions function,and in UV protection plants, flower pigment formation play a promoting role.The resistance of Amorpha fruticosa L. and nodules form has important relationship.The regulation of flavonoids is important link for the process of formation of root nodules.In recent years,there are many researchs on Amorpha fruticosa L.physiological characteristic.But rarely news on the extraction of flavonoid compound from the leaves of Amorpha fruticosa L.In this experiment, Amorpha fruticosa L. leaves as the main raw material.Using advanced extraction method, reasonable design of experiment, scientific analysis of the results. To determine the content of flavonoids in Amorpha fruticosa L. leaves, optimize of the extracting method process parameters, optimization of macroporous resin purification process parameters and preliminary identification of the extraction. The main experimental results are as follows:(1) Ultrasonic method optimizing process parameters:liquid-to-solid ratio, extraction of power, ethanol concentration, extraction time and extraction times were selected as the single factor, by orthogonal experiment and response surface experiments for two kinds of models for verification, common optimizing process parameters. The optimal conditions: liquid-to-solid ratio 50ml/g, extraction of power 316W, ethanol concentration 50%, extraction times 4.The content of flavonoids is 66.43 mg/g. Experimental factors on flavonoids extraction effect with strong and week relationship:extraction times> ethanol concentration> liquid-to-solid ratio> extraction of power.(2) Microwave water bath method optimizing process parameters:ethanol concentration,extraction time, extraction times,bath temperature and liquid-to-solid ratio were selected as the single factor,by orthogonal experiment and response surface experiments for two kinds of models for verification, common optimizing process parameters. The optimal conditions:ethanol concentration 60%, extraction time 6 min, extraction time 60 ml/g,bath temperature 80 ℃. The content of flavonoids is 72.09 mg/g. Experimental factors on flavonoids extraction effect with strong and week relationship:ethanol concentration> bath temperature> liquid-to-solid ratio> extraction time.(3) Macroporous resin purification optimizing process parameters:With the total rate of purification as indexes,under the static absorption and desorption,to optimzie choosen the type of macroporous resin and the ethanol concentration of dissolve the crude extracts from flavones; under the dynamic absorption and desorption, based on single factor, studying the inflution of sample concentration、sample flow rate% sample volume、sample PH、the ethanol concentration of elution、the rate of elution to the total rate of purification,and find the optimal technology conditions.As results, with 10% ethanol dissolve the crude extraces, D101 macroporous resins separate and purify the sample solution. Optimum technology:sample concentration 1.5mg/ml,sample flow rate 3ml/min.sample volume 100ml, sample pH=4, the ethanol concentration of elution 70%, the rate of elution 3ml/min. The technical condition is scientific and reasonable that can be used to separate and purify flavones of Amorpha fruticosa L.leaf. The flavonoids content of the crude extracts gained under these conditions was 56.2%, and 2.9 times more than that of the original sample 18.9%.(4) Preliminary identification results:Using the color reaction and uv spectrum characteristics identification the flavonoids of Amorpha fruticosa L. leaves. Determine the main types of flavonoids:flavones and flavonols(3-OH replace).