Study On Extraction Separation And Purification Of Apigenin From Viola Yedoensis Makino

The thesis had described the content of apigenin in Viola yedoensis Makino and several kinds of Violaceae at the different seasons around Zhangjiajie campus of Jishou University. And extracting efficiency of apigenin in Viola yedoensis Makino had been studied in the conditions of water bath,soxhlet extraction,microwave-assisted extraction,supersonic extraction and supercritical CO₂ extraction.The factors which effect extracting of apigenin in Viola yedoensis Makino had been studied by L₁₆(4⁵) orthogonal experiment,such as the solvent,concentration,solid/liquid,temperature and extraction time.The best extraction condition of apigenin in Viola yedoensis Makino was chosen by the results of analysis of variance.The best macroreticular resin to purify apigenin was chosen by the experiment of static adsorption,dynamic adsorption and de-adsorption.Determining the extractions from the Viola yedoensis Makino and apigenin standard with HPLC and UV,establishing measure method of the apigenin content in Viola yedoensis Makino.Results showed that the optimum condition of extraction was that the concentration of methanol was 70%,the ratio of solid to liquid was 1:10,the extracting temperature was 50℃,the extracting time of ultrasonic was 30 mins.LSA-5B resin and column chromatography silica gel were the better for isolation & purification of apigenin in Viola yedoensis Makino.The optimum conditions of supercritical CO₂ extraction were as following:solid/liquid 5:3,extraction pressure 20 MPa,extraction temperature 55℃,extraction time 2 h,the temperature of separatorâ… 55℃,and the pressure 9 MPa,the temperature of separatorâ…¡40℃,and the pressure 5 MPa.There was a good linearity between peak areas and injection quantity of apigenin within the range of 1.58～15.80mg.L^-1,r=0.9996;And Utilizing the measure of UV at 345nm there was a good linearity between the absorbance and the quantity of apigenin within the range of 3.33～19.98μg/mL,r=0.9999.The highest content of the apigenin in the Viola yedoensis Makino was 0.0236%,which is harvested in spring.The contents of apigenin in the four kinds of Violaceae are respectively 0.0221%in Viola philippica Cav.(Viola yedoensis Makino),0.0144%in Viola rossii Hemsl.,0.0137%in Viola selkirkii Pursh., 0.0120%in Viola confuse Champ;The content of purification apigenin reached 96.8%.