Study On The Quality Of Lindleyanum DC And Separation And Purification Of Eupatorium Lindleyanum DC Lactone

Eupatorium LindLeyanum DC is a natural herb of distinguished efficacy for relieving cough and asthma. Eupatorium LindLeyanum DC Lactone separated from it has an effect of anti-nasopharyngeal carcinoma. Study on the pharmacology and utilization of Eupatorium LindLeyanum DC is not yet mature. The quality of LindLeyanum DC is studied and the optimal technology of isolation and purification of eupatorium LindLeyanum DC Lactone is established as well.The whole Experiment is divided into three parts:1. Study on the TLC and RP-HPLC behavior of Eupatorium LindLeyanum DC Lactone A and Hyperin mentioned in the Chinese Pharmacopoeia, LindLeyanum DC Lactone B was also researched, to establish a complete and accurate, simple quality control methods of LindLeyanum DC. Based on this, LindLeyanum DC from Six different areas were studied, and provided a good way to learn the quality of LindLeyanum DC and choose the raw material for the the separation and purification of Eupatorium LindLeyanum DC Lactone.2. Two different extraction methods such as reflux extraction and ultrasonic extraction were used to study the best conditions for exracting Eupatorium LindLeyanum DC Lactone A, Eupatorium LindLeyanum DC Lactone B from LindLeyanum DC. The results showed that:inspected the effect of five factors (extraction temperature, extraction time, the ratio between solvent and dreg, extraction times, extraction solvents) to do the single factor experiments. In analysis of orthogonal experiments, the optimal technological conditions of reflux extraction are as follow:90%methnol solution as extracting reagent, proportion of materials and extracting reagent was1:20, extraction temperature is70℃, extraction time is2.5hours, the number of extraction reach the highest rate are0.0168%,0.0394%。For the ultrasonic extraction, inspected the effect of four factors (extraction time,extraction temperature, extraction times, extraction solvents), based on this, established orthogonal experiments, the results suggested that the optimal technical conditions were as follows:90%methanol solution as extracting reagent, proportion of materials and extracting reagent was1:20,30min extraction time for2times, and the yield were0.0182%and0.0421%. Two extraction method is compared, ultrasonic extraction is more superior than reflux extraction, for its higher extraction rate and stable characteristic and lower costs.3. Study on the purification process of Eupatorium LindLeyanum DC Lactone A and Eupatorium LindLeyanum DC Lactone B, the best purification process was set up. Petroleum ether, chloroform, water were used to extract polarity parts contained Eupatorium LindLeyanum DC Lactone. Tested by TLC and HPLC, Eupatorium LindLeyanum DC Lactones are mainly concentrated in the chloroform layer. Recovery chloroform solvent and extracts were gotten. Various column chromatographic were used to study on the purification of Eupatorium LindLeyanum DC Lactone A and Eupatorium LindLeyanum DC Lactone B. Considering the operations, production cost, and other factors, the best purification technology was determined. Using dry column chromatography, methanol and chloroform mixture solution as eluting agent, TLC and HPLC were used to identify the aim content. Recovery concentrate contains Eupatorium LindLeyanum DC Lactone, and using column chromatography to purity the target parts again. Middle-pressure liquid chromatography with RP-C18was used under1:1of methanol-water mixtures as eluent, the purity of the standard substances were reach to85%or more. Normal phase chromatography was used to get the products to a purity of90%. Recording to the amount of the products, the Preparative high performance Liquid chromatography or Half-preparative high performance Liquid chromatography were selected.