Studies On Preparation And Purification Of Water-soluble Starfish Glycosides From Asterias Rollestoni And Some Biological Activity Examination

Starfish glycosides are crude marine materials which have widely bioactivities and pharmacological activities. People often use dilute alcohol to get glycosides from starfish. The excess use of organic solvent would not only raise the cost in industrial production but also pollute environment seriously. Futhermore, the glycosides extracted by organic solvent have bad water solubility so that the approach and extention of applications in the fields of clinic medicine and agriculture are largely limited . It has not been reported on the preparation and purification of water-soluble starfish glycosides and on their bioactivities. In this study, water-soluble glycosides from Asterias rollestoni are prepared and purified by the methods of water extraction, macroporous resin column ,. silica gel column and Sephadex LH-20 column. We do some research on their antifungal and antitumor activities and get a pooled water-soluble glycosides single component with high antifungal and antitumor activities.The results of property identification show that the crude water-soluble starfish glycosides we get by the methods of water extraction and macroporous resin column eluted by 70% alcohol are steroidal glycosides. The results of antifungal activities examination show that the crude water-soluble starfish glycosides have distinct antifungal activities against six fungi Aspergillus niger, Candida albicans, Saccharomyes cerevisise, Schizosaccharomyces probe, Fusarium oxysporumf.sp.cucumerinum, Colletotrichum gloeosporioides. The minimum inhibitory concentration (MIC) are at 0.625, 0.625, 0.313, 0.156, 1.25 and 0.625 mg/mL respectively. The results of antitumor in vitro activities examination show that the crude water-soluble starfish glycosides have distinct antitumor activeties against the tumor cells HeLa, A-549, SGC-1901 and Bel-7402, and the IC50 are at 14.66, 15.37, 13.44 and 12.74 g/mL respectively.Loaded on a silica gel column, the crude water-soluble starfish glycosides are purified into four fractions, which are designated as pSF-1, pSF-2, pSF-3 and pSF-4 respectively.The results of antifungal activities and antitumor activities examination in vitro show that the fractions of pSF-3 and pSF-4 have distinct antifugal activities. The results of antitumor activities show that the IC50 of the pSF-3 against HeLa, SGC-1901, A-549 and Bel-7402 are at 7.05, 5.28, 5.32 and 6.16 g/mL respectively. And the IC50 of the pSF-4 against HeLa, SGC-1901, A-549 and Bel-7402 are at 7.24, 5.94, 5.24 and 6.21 g/mL respectively.By the methods of Sephadex LH-20 column, we get the purified component SF-3 from the silica gel column fractions. The results of high pressure liquid chromatography (HPLC) show that the purity of SF-3 is higher than 96%,which demonstrates that SF-3 is the single component. The results of antitumor activities examination in vitro show that SF-3 can distinctly inhibit the growth of HeLa and Bel7402 tumor cells. The results of antitumor activities examination in vivo show that SF-3 can distinctly inhibit the neoplasia of S180 and the inhibition rates are 23.34%, 36.33% and 46.53% at the doses of 2, 5 and 10 mg/kg B.W. SF-3 can also enhance the thymus and spleen exponent of S180 mice at the dose of 10 mg/kg B.W.In summary, we utilize the methods of water extraction, macroporous resin column, silica gel column and Sephadex LH-20 column to prepare and purify water-soluble starfish glycosides. We get two purified components pSF-3 and pSF-4 which have distinctly antifungal and antitumor biological activities and a single component SF-3 which has distinctly antitumor biological activities. This study lays solid foundation to the development and research of highly bioaictive natural antifungal and antitumor marine medicaments.