The Purification Of Main Active Components From Luidia Quinaria Von Martens And The Bio-activity Study Of Asterosaponin

Luidia quinaria von Martens, belonging to the family Echinodermata, Asteroidea, is a kind of marine invertebrate.It is widely distributed along the Huanghai Sea and Bohai Sea, especially in Jiaodong peninsula.To make full use of Luidia quinaria von Martens in medicine and food, the preparation, structural determination and biological activity study of asterosaponins (as the key section), together with the determination of polyunsaturated fatty acids and primary nutritional components, the extraction and purification of acidic mucopolysaccharides from Luidia quinaria von Martens in Huanghai Sea were systemically and fully investigated in this study.Luidia quinaria Von Martens were firstly extracted by a series of classical solvent methods and then became crude asterosaponins.The purification steps were based on macroporous resins adsorption column chromatography, silica gel column chromatography, sephadex LH-20 column chromatography, reversed-phase HPLC, finally the pure chemicals were got.Based on the UV, IR, NMR and HPLC-MS spectra, the components of asterosaponin were proved to be steroidal saponins and quite complex.The specific structural confirmation needs further research.From the determination findings of the nutritional compontents, the content of crude fat is 2.71%.Refined fatty acids were successfully extracted and refined using solvent extraction and silica gel column chromatography with yields of 2.14%.The gas chromatography analysis shows that polyunsaturated fatty acids content in the refined fatty acids exceed 55%, and the EPA content exceeds 45% .Acidic mucopolysaccharides were extracted by sodium-hydroxide method and enzyme-decomposition method.The gross products were then deproteined by trichloroacetic acid and twice enzymic hydrolysis and then made as acidic mucopolysaccharide fractions.The fractions were further separated and purified by ion-exchange chromatography and Sephadex, and finally analyzed with Sephadex.The identification results show that fractions are composed of only one component combined with sulfate.The average molecular weight is about 12 kD.The results of phyarmacological study of refined asterosaponin show it has many biological and medical properties, such as hemolytic activity to rabbit erythrocytes at 0.1mg/mL, obvious cytotoxicity towards SMMC-7721 and PC-3M in certain dose-dependent manners, antitumor activity on mice transplanted tumor cells of U14 cervical carcinoma and Lewis lung cancer in good dose-dependent manners. The HI of refined asterosaponin is 9 950 and the concentration is 25μg/ml. In conclusion, Luidia quinaria Von Martens are good sources of many bioactive substances , such as asterosaponins, polyunsaturated fatty acids and acidic mucopolysaccharides, which can be separated and purified with modern biochemical technologies.Function evaluations show that asterosaponins have biological and medical activity in hemolytic,cytotoxicity towards normal cells and antitumor.