Isolation And Characterization Of Constituents From Halocynthia Roretzi Drasche Shell And It’s Biological Activities

Halocynthia roretzi Drasche belonging to the Chordates, Urochordata, class Ascidiacea, is an abundant starfish distributed in Bohai Sea and Huanghai Sea. The study contained four parts: the extract process from H. roretzi Drasche shell,isolation and purification of extraction, structure identification of compounds and biological activity(antibacterial, antioxidant and anti-tumor experiment).H. roretzi Drasche was exhaustively extracted by 60%, 75%, 95% hot ethanol(each 2 h), respectively. The extraction with 75% ethanol yielded up to 10.40%. Vanillin-sulfuric acid colorimetry showed that the saponin extraction rate was 14.3%. LB reaction of the shell extraction was positive, and foam experiments also indicated that the extraction contains steroidal substances.The extraction with 75% hot ethanol was seperated three fractions, PEE, ETA and BUA, extracted by petroleum ether, ethyl acetate, n-butanol in turn. Using a variety of separation means, including thin layer chromatography, silica gel column chromatography, preparative thin layer chromatography(p TLC), Sephadex LH-20 gel column chromatography and recrystallization, PEE, ETA and BUA were separationed and purificationed, respectively. HRA and HRB were isolated and purified from petroleum ether layer, and identified by their physical and chemical properties, spectral analysis and documentation control method, known as diethyl 24-[(Z)-3-methy-1-hexenyl]-heptatetracontanedioate and cholesterol. HRC was isolated and purified from BUA, and identified as 6-(2,,3,-dihydroxy-4,-hydroxymethyl-tetrahydro-furan-1,-yl)cyclo-pentadiene [c] pyrrole-1,3-diol. HRA and HRC were first obtationed from H. roretzi Drasche shell.The antibacterial activities data of different polarity reagent extracts from H. roretzi Drasche shell showed that the ETA inhibited the growth of Escherichia coli, Clostridium perfringens, Shewanella sp., as well as Bacillus subtilis, and significantly inhibited the growth of Staphylococcus aureus; other extracts to the five bacteria had different degrees of inhibition.Scavenging effects on O2-?,?OH and DPPH? of different polarity reagent extracts from H. roretzi Drasche shell were in a concentration-dependent manner. Generally speaking, the ETA and BUA showed higher antioxidant activity.Anti-tumor activities of the extraction of H. roretzi Drasche shell extracted with 75% ethanol were studied. Three kinds of tumor tumor cells(AGS cells, Hep G2 human hepatoma cells, SKOV3 ovarian cancer cells) were chosed for anti-proliferation activity experiment, then detected with CCK-8 kit. The data showed that with the concentration of total extract increased gradually, cell proliferationi rate was decreased, which indicated that the total extract for three kinds of cancer cells have toxic effects. Among these cells Hep G2 cells were more significantly inhibited. The alcohol extract of H. roretzi Drasche shell as an active ingredient of anticancer drugs could provide a theoretical basis for further research and development of marine drugs.