Studies On The Quality Standard Of Eupatorium Lindleyanum Dc. And Preparation Of Eupalinolidea

Eupatorium LindLeyanum DC. as a kind of asteraceae plant, is a special natural herb in XuYi county of JiangSu province. It has the efficacy of detoxification, eliminating phlegm and stopping cough, dieresisdetumescence and reducing the blood pressure when it was used as medicine with the whole plant. It is commonly used to cure diseases such as chronic bronchitis, bronchitis and other respiratory system diseases as well as hypertension in clinic. Foreign scholar had confirmed that the eupalinolide (A, B, C, D) they had isolated from Eupatorium LindLeyanum DC. had the resistance of KB cell in pharmacology experiments in1979. The study of eupalinolide A is extremely little these days, the paper is not only established the quality standard of Eupatorium LindLeyanum DC., but also determined the optimal process of extraction, isolation, and purification for eupalinolide A. The study achievements of this paper are following:1. Established the quality standard of Eupatorium LindLeyanum DC.. The paper established a set of analytical method through the TLC and HPLC behaviors of eupalinolide A and eupalinolide B. The analytical method is simple, accurate, fast and economic. It provided a practical program for the quality standard research. In addition, the experiment determined the content of the six batch medical material from different regions. It not only offered the theoretical basis for the medicinal material quality, but also supplied the basis of selection and detection method for the subsequent work.2. The study confirmed the best isolation method of eupalinolide A by means of organic solvent extraction and ultrasonic extraction. The method has advantages of high extraction efficiency and low costing. During the solvent extracting process, five main factors including solvent, time, temperature and times of extracting as well as the ratio of solid to liquid were studied by using single factor analysis method. Then the technological conditions for eupalinolide A extraction are optimized by orthogonal experiment:90%methanol aqueous solution as extracting solvent, ratio of solid to liquid is1:20, extracting temperature is70℃, extracting time is2.5h. During the ultrasonic extraction process, four main factors including solvent, time, times of extracting and ratio of solid to liquid were researched by using single factor analysis method. Then the process conditions for eupalinolide A extraction are optimized by orthogonal experiment: 90%methanol aqueous solution as extracting solvent, ratio of solid to liquid is1:20, extracting time is30min, the number of extracting times is2. Comparing these two methods comprehensively, ultrasonic extraction process is superior to solvent extracting process in some ways like extraction rate and stabilization.3. The paper confirmed the best isolation and purification method of eupalinolide A through the research of process conditions. Firstly, we used aether petrolei and ethyl acetate to extract the Eupatorium LindLeyanum DC. extractum successively in the preliminary isolation. Secondly we determined that eupalinolide A was mainly existed in the part of ethyl acetate by analysis of HPLC. Thirdly, recovered the ethyl acetate, utilized the silicagel column to make the ethyl acetate part in several sections by different polarity. Through the HPLC behavior, we detected that eupalinolide A was mainly existed in the eighth part of ethyl acetate, then recovered it and obtained the extractum. Comparing different chromatographic column’s affection for isolation and purification of eupalinolide A, and considering the cost,cycle of production, we confirmed the optimal method:the Fr.8is subjected to a medium-pressure MCI column, eluting by a MeOH-H2O4:6to obtain the component that contain eupalinolide A. After removal of MeOH-H2O under reduced pressure, the residue is obtained, which is fractionated by silicagel column chromatography eluted with CHCl3-MeOH or petroleum ether-acetone, detect the target part with TLC and HPLC, then recover the target part under reduced pressure. This part will be purified by a medium-pressure RP-C18column eluted with MeOH-H2O4:6, collect the eluant500ml each time, then detect the target part with TLC and HPLC, recover the target part under reduced pressure, eupalinolide A will separate out in the form of white sediment, finally, we can use crystallization and recrystallization process to obtain the eupalinolide A, which its purity can reach at98%.