Study On Extraction And Purification Of Herba Artemisiae Scopariae And Its Evaluation By The Method Of "Pharmacodynamics Combinded With Finger Print"

Herba Artemisiae Scopariae ,which is adducted by Chinese Pharmacopeia, can be ofen seen in liver-protecting and cholagogue complex prescription. Presently it is usually extracted by water as tradition, and sometimes processed by "water extracted—alcohol precipitated" technology or percolation technology in Chinese drugs pharmaceutics, but research works are seldom done to evaluate the above-mentioned process.With the development of science and technology, the innovation of pharmaceutics technology and its serious evaluation is very important to the elevation of technique level and elucidation science connotation of Chinese formulated products.The commonly used technology evaluating system, with reservation rate of single ingredient in the herb as the parameter, is limited in the evaluation of multi-components Chinese herbs and the evaluating method of general effective regional is indefinite. The development of finger printing technology provides a more exquisite index to the evaluation of reservation of pharmacodynamic action substance in Chinese drugs pharmaceutics technology. Even so, rigorous researchers still have reasons to doubt that if the application of new technology keeps the drug' s action while keeping the correlated components in finger printing. Therefore, the method of "finger print combined with pharmacodynamics" as index which accompanies and evaluates new pharmaceutics technologies is more persuasive.This article explores the application of macroporous resin in theextrcacting of Herba artemisiae scopariae , and scientific evaluation of the purification is also be done in the research. It has established the finger print of Herba artemisiae scopariae and investigated the influencing factor of chlorogenic acid in the extract to aim directly at the research foundation of Herba artemisiae scopariae , and studied the extraction process and purification process by macroporous resin with the reservation rate of the main peak in finger print of Herba artemisiae scopariae and the pharmacodynamics evaluation as indexes. The main contents and results are as followings:1. The establishment of finger print of Herba artemisiae scopariaeThe finger print of Herba artemisiae scopariae is established with reversed phase high performance liquid chromatdgraphy and the corresponding methodological research is undertaken to evaluate the quality of Herba artemisiae scopariae from different habitats and batches.2. Influencing Factors of Caffeotannic Acid' s Stability in Extracts of Herba Artemisiae ScopariaeThe Herba Artemisiae Scopariae decoctions by water or alcohol is heated in different pH and for different period, and the content of chlorogenic acid is determined. The result shows that the heat temperature and pH of the decoctions are important factors which influence stability of chlorogenic acid, and alcohol system is in favor of its stability. We should pay attention to these factors in the extracting, recovering the solvent, concentrating and drying process of Chinese drugs pharmaceutics which have the content of chlorogenic acid.3. The extraction process of Herba Artemisiae ScopariaeWith chlorogenic acid, peak No. 8 and peak No. 13, which stand for different polars in the finger print of Herba Artemisiae Scopariae, as the indexes, the different methods are comprehensively scored based on the extraction rate of the three compositions. After the extraction times is determined, the parameters of the extraction process of Herba Artemisiae Scopariae are examined with LOO orthogonal design. Then the chosen optimized process is compared with the traditional water-extraction process with the pharmacodynamics research of resisting acute liver injury. The results show that the optimized process parameters are the extraction of Herba Artemisiae Scopariae with 10 times, 8 times and 8 times of 80%alcohol for 3 times, 1 hourper time. The comparison of ALT, AST and pathological indexes show the trend that the alcohol-extraction process is better than water-extraction process. 4. The purification process of Herba Artemisiae Scopariae decoction with macroporous resinIn this study, the influencing factors of static adsorbting process of extract of Herba Artemisiae scopariae with macroporous resin are studied at first. With the absorbances and the adsorption quantity of caffeotannic acid as indexes, the adsorption effect of different macroporous resin and timetable of static adsorption are investigated. The influence tendency and extent of density and pH of the physic liquor in the static adsorption by use of factorial experiment are investigated. With the HPLC finger print of extract of Herba Artemisiae scopariae as an index, the adsorption effect is investigated. The result shows that the adsorption effect are different with different types of macroporous resin , and the adsorption equilibrium time is 6 h;Density and pH of the physic liquor are important factors of adsorb. The results may offer some information for the determination of refinement process of Herba Artemisiae scopariae decoction with macroporous resin.Based on the results above, with the HPLC finger print of extract of Herba Artemisiae scopariae as indexes, the kinesis adsorb and elution process with quantity of caffeotannic acid are investigated and the maximum quantity of physic liquor and the type and quantity of elution solvent are determined. The results show that the maximum quantity of caffeotannic acid that each gramma of dray resin can adsorb is 20. 9mg, and the elution solvent determined is five times column volume of 80% alcohol;The verification experiment indicats that this process decreased the yield of extract to 2. 5%, and meanwhile it retained the whole components of the HPLC finger print effectly. So the Refinement process can enrich the active components in Herba Artemisiae scopariae effectively.5. Evaluation to the refinement process of extract of Herba Artemisiae Scopariae with Macroporous resin by the method of "pharmacodynamics combinded with finger print"The refinement process of extract of Herba Artemisiae Scopariae with macroporous resin is evaluated with the reservation rate of main peak in the finger print as an index. The CCL, is injected to mouse to establish a model of acute hepatic injury, then the activity of ALT and AST in the serum isdetermined and the pathological diversity of the histological section is observed to compared the hepatoprotective effect of the refinement product with extract of Herba Artemisiae Scopariae. The results show that before or after the refinement, the finger prints of the extract of Herba Artemisiae Scopariae are almost the same, the hepatoprotective effects have no significant deviation and no obvious acute toxic reactions are observed.The results above show that the rate of extract can be decreased and the drug contents of the preparation can be increased while the main peak of the finger print and the hepatoprotective effects can be reserved by optimizing the extraction process and the purification process with Macroporous resin. This research can provide proof for reference for the use of Herba Artemisiae Scopariae in complex prescription of Chinese drugs pharmaceutics.