Isolation, Identification And Biological Activity Of Chemical Substances Extracted From Anthocidaris Crassispina Shell

Sea urchin shell is the application site, and it contains lots of marine active substances,with a wide range of medicinal value and health effects, herbs were called "sea urchin."Modern medicine proved that’’sea urchin taste salty, flat. Its functions are hlegming, swelling,relieving pain, and detoxification.’’ Currently, lots of sea urchin shells are discarded as waste,it is not only a waste of potential medicinal resources but also put pressure on the environment.The study of its active ingredient is imminent.Dalian common sea urchin shells were regarded as raw material for research in thisexperiment, the study contains four parts: the extract process of Anthocidaris crassispina shelland Hemicentrotus pulcherrimus shell, isolation and purification of Anthocidaris crassispinashell, structure identification of compounds and biological activity (antibacterial andanti-tumor experiment).Using ultrasound-assisted method, heat and cold extraction method for the preparation ofthe extract, and it was extracted with75%ethanol, and concentrated to give extract. It wasfound that the extraction rate and steroids content with ultrasound-assisted is highest, followedby the heat method. And the extraction rate and the active ingredient content of Anthocidariscrassispina shell were higher than Hemicentrotus pulcherrimus shell.The extract was extracted with petroleum ether, ethyl acetate and n-butanol, andthree parts were got: PEE, ETA and BUA. Using a variety of separation means, for examplethin layer chromatography, silica gel column chromatography, preparative thin layerchromatography (pTLC) and recrystallization, petroleum ether (PEE) and n-butanol layer(BUA) extracts were separationed and purificationed, respectively. Compound ACB wasisolated from petroleum ether (PEE), its structure was identified by physicochemicalproperties, spectral analysis and documentation control methods, and a known compoundpalmitic acid was identified. It is worth mentioning that it is the first time for ACB to isolatedfrom Anthocidaris crassispina shell. There compounds were isolated and purified fromN-butanol layer (BUA), and they were named as ACA, ACT and ACS. ACA has beenidentified as a known compound, and its structure is (3β)-cholest-5-en-3-alcohol, which isthe first time for ACA to isolated from Anthocidaris crassispina shell. The structure ofcompound ACT and ACS are more complex, and they are possible for the newcompounds.Then the work about structure identification is still being studied.This antibacterial activity and anti-tumor activity about various extracts of theAnthocidaris crassispina shell were researched. It was found from antibacterial experimentsthat anthocidaris crassispina shell extract had good antibacterial activity. Anti-tumor experiments about ACT, ACS showed that compound ACS did not have cytotoxicity,compound ACT showed a significant cytotoxicity on K562, HL60, SKBR-3, MCF-7andSW620cell, and with the concentration of compound ACT gradually increasing, the apoptosiscontent was gradually increased, when the concentration of ACT was150μg/ml, thepercentage content of apoptosis got the highest levels. Therefore, it was refered that compoundACT had significantly cytotoxicity and induced apoptosis, and it can become an activeingredient of potential anticancer drug. However, the cell structure and mechanism aboutcytotoxicity and the induction of apoptosis were unclear and pending further study.