Research On Alkaloids From Fritillaria Thunbergii Flowers:Extraction, Purificationand Effectiveness Evaluation

Fritillaria thunbergii is a traditional Chinese medicine with kinds of effects. In recent years, it have been found that the bulbs and flowers contain similar main components and have same phamacological effect. On this study, the raw materials were flowers of Fritillaria thunbergii, with different methods of extraction, separation, purification to obtain the components. And further study on its active ingredients was studied in this paper in order to get the most suitable methods. The research lays foundation for development an utilization of Fritillaria thunbergii flowers. The major results as follows:1. Extract from Fritillaria thunbergii flowersThe extract from Fritillaria thunbergii flowers was evaluated by means of reflux extraction, ultrasonic extraction, microwave extraction, respectively. The factors like ethanol concentration, liquid-solid ratio, temperature was taken consideration in the research. After response-surface experiments, the result of reflux extraction were as this:the optimization model P<0.0001, the degree of lack of fit was insignificant, the coefficient of determination R2=0.9709, the correction coefficient RAdj=0.9419, the extracting amount was 0.0672mg/g. The result of ultrasonic extraction were as this: the optimization model P<0.0001, the degree of lack of fit was insignificant, the coefficient of determination R2=0.9755, the correction coefficient RAdj=0.9510, the extracting amount was 0.1287mg/g.The result of microwave extraction were as this: the optimization model P<0.0001, the degree of lack of fit was insignificant, the coefficient of determination R2=0.8543, the correction coefficient RAdj=0.7086, the extracting amount was 0.1163mg/g. Results shows that the ultrasonic extraction is the most suitable. The optimum ultrasonic extraction process of alkaloids for Fritillaria flower is as follows: ethanol concentration 79%,ultrasonic power 280 W, ultrasonic time 47 min, liquid-solid ratio 31:1. Under this condition,the forecast amount of Fritillaria flower extract alkaloids is 0.1304mg/g and the actual extraction is 0.1287mg/g.2. Purifying after the crude extractBased on the extraction and separation of Fritillaria thunbergii Mid flower, the bestpurification process of total alkaloids from Fritillaria thunbergii flower was selected by macroporous resin. Optimum macroporous resin and its purification process was selected by static and dynamic adsorption and desorption kinetics test. Optimal purification process conditions were: optimal macroporous resin type HPD100, the concentration of sample solution 0.5g/mL with pH as 8.0, optimal sample volume 40 mL, flow rate of adsorption1.5BV/h, washed by 1BV 80% ethanol, flow rate of desorption 2BV/h. Under this optimized technology, the content of total alkaloids was more than 60%. This method can obviously improve the total alkaloids content in extract of Fritillaria thunbergii Mid flower.3. Conducting effective assessment on the purificationThe antitussive effect were investigated though the experiments of using Ammonia leading to cough and Sulfur dioxide leading to cough respectively. The result shows that,compared with negative groups, high does group of extracts from Fritillaria thunbergii flowers had extremely significant antitussive effect with its R%>130%. It confirmed Fritillaria thunbergii flowers had significant antitussive effect. The expectorating sputum effect were investigated though the experiments of Phenol red expectoration drainage. The result shows high does group of extracts from Fritillaria thunbergii flowers had extremely significant expectorating sputum effect compared with negative groups. It confirmed Fritillaria thunbergii flowers had significant expectorant effect.