Optimization Of Extraction And Separation Purification Methods And Assaying For Phytoestrogen-genistein

Objective:Research to best extraction, separation and purification route of genistein in Sophora (genistein GST), and determination of its content. Methods:To combine single factor and orthogonal test, to investigate every factors of influenced extraction rate of genistein rough products through single factor experiment, then to optimize each technology parameter of extraction methods by orthogonal experiment, determined the best extraction method through comparison3species extraction methods (heating reflow law, ultrasonic law, and microwave law). To compare3species separation purification methods (heating reflow law, ultrasonic law, and microwave law) by extraction rate and purity of genistein, established the best extraction and purity method, optimizing every process parameters of separation and purification by single-element and orthogonal experiment, and finally assaying of genistein in Sophora by high performance liquid chromatography (High performance liquid chromatography HPLC). Results:The best extraction method is ultrasonic extraction law, second for microwave extraction law and heating reflow extraction law through comparison. Optimum process parameters of ultrasonic extraction method: ethanol concentration85%, solvent usage25ml/g, ultrasonic extraction time40min, ultrasonic power300W, ultrasonic extraction temperature55℃, for2times. Average yield of genistein is135.64mg/g by this method. Large hole resin adsorption law is the best separation purification method, solvent crafts extraction and recrystal law haven’t markedly difference. The technology parameter of large hole resin adsorption law has optimization, Best separation and purification technique of macroporous resin adsorption parameters:XDA-200resin,25℃,1.5mg/mL sophorae genistein crude solution adsorption column on12h by1mL/min flow concentration. When the sample is1BV, using the3BV95%ethanol solution at65℃,1mL/min flow rate elution, through this method of purification, average rate of829.07mg/g and average purity of90.13%. Using the HPLC method for the determination of genistein in Sophora content, genistein derived from113.50mg/g by the best routes. Conclusions:Through system comparison, we cleared advantages and disadvantages of several extraction and separation purification methods, obtained a best extraction, and separation purification technology route, offered basic of chemical data for genistein of economic efficient extraction, and separation purification, through effective development of Huai corner medicinal components and production biological preparations of high added value, has important economic benefits and social benefits; clear content range of genistein in Huai corner by content determination, expanded the sources of raw materials of genistein, Sophora resources development and full and reasonable use of medicinal value, effectively promoting the development of Sophora processing industry and pharmaceutical companies.