Study On Extraction And Purification Processes Of Total Of Opuntia Saponins

Opuntia dillenii Haw. is a plant belonging to genus Opuntia in Cactaceae family. The plants of Opuntia have a wide distribution in the world. Recent studies show that total of Opuntia saponins is an important biologically active component in genus Opuntia plant. There is no systemic report on the research of extraction and purification technique of total of Opuntia saponins in China. The objective of this paper is to lay a foundation for compositive utilization and exploitation of Cactaceae. On the bases of previous research of total of Opuntia saponins and the facilities of the laboratory, this thesis systematically studied a colorimetric method for quantitative analysis of total of Opuntia saponins , optimized the parameters of ethanol reflux extraction which was screened by orthogonal design. The purification processes of total of Opuntia saponins with D101 macroporous resin was generally studied by taking adsorption quantities, eluting ratio and so on. The major results are shown as follows:1. The content of total of Opuntia saponins was determined by UV-VIS spectrophotometry after it has been colored. Using oleanolic acid as a standard substance, assured detective wavelength of total of Opuntia saponins was 548 nm and standard curve equation was A=6.425C-0.0016, the average recovery was 98.47% with RSD of 3.29%. The method is rapid, accurate and repeatable.2. Ethanol reflux extraction processes on total saponins from Opuntia were studied. Single element test and L₉(3₄) orthogonal design test were performed about alcohol concentration, ratio of solid to liquid, extraction temperature and time. As a result, the optimal conditions were obtained as follows: the solvent is 80% ethanol, the temperature is 70℃, the extracting time is 6h and solid-liquid ration is 1: 15. The result shows that the average extraction rate of total of Opuntia saponins is 0.1159%.3. The separation and purification of total of Opuntia saponins were carried out with the method of macroporous adsorption resin. The absorption and desorption of D101 macroporous resin to total of Opuntia saponins were studied. The result indicated that D101 macroporous resin fits for the separation of total of Opuntia saponins and its static state adsorption ratio reaches 9.922mg/g dried resin. The optimal purification parameters of D101 were: pH of 3, flowing rate of adding sample and adsorption solution were 2BV/h and 2BV/h respectively. Through response orthogonal test and statistic software STATGRAPHICS plus 5.1 and variance analysis, the optimization operational condition was established. The ratio diameter to high of the chromatographic column was 1:11.57, and temperature was 28.83℃. It is desired to elute with 5BV at the flow rate of 2BV/h then desorpt with 5BV and 50% ethanol at the same flow rate, to effectively separate total of Opuntia saponins.4. Content of total of Opuntia saponins of 8 fermentation Opuntia analysed by optimal technique flow.To sum up, the results of the whole experiments demonstrated that techniques such as ethanol reflux extraction and macroporous resin purification could effectively enhance the extraction and purification rates of total of Opuntia saponins.