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1、respectivelyandthehostwasforward.Lastly,3.4610mgofactiveingredientwasobtainedfrom1gCanavaliamaritimeatpurityof99.48%.Thestructureofisolatedcompoundwasidentifiedtobe(2R,3S,4R,5S)-2,3,4-Trihydroxy-6-methoxy-6-oxo-5-[(3,4,5-trihydroxybenzoyl)oxy]hexanoicacidwithamolecularweighto
2、f376.05byHPLC-MS,1H-NMRand13C-NMR.Theisolatedpureproductshowedstrongantioxidantactivityandthescavengingrate(IC50)for·OH,O2-·andDPPHradicalswere0.052,0.057,0.010mg/mLrespectively.CaulerpinwassuccessfullyextractedandseparatedbymeansofMAEcombinedwithHSCCCfrompinnatefernalgal.The
3、optimizedextractionconditionsofcaulerpinobtainedbyorthogonalexperimentsincluded90%ethonalasextractionsolvent,microwavetemperatureof40℃,microwavepowerof300W,microwaveradialtimeof15minandmaterial-liquidratioof1:40g/mL.Undertheoptimalextractionconditions,throughduplicatedexperim
4、enttheyieldofcaulerpinis92.93%,RSDis0.50%.Caulerpinwasseparatedwithatwo-phasesolventsystemcomposedofn-hexane-methanol-water(2:1:1,V/V)fromcrudeextract.ThespecificparametersofHSCCCseparationwereasfollows:photogenicasthestationaryphase,lowerphaseasthemobilephase,forewardrotates
5、peedof850rpm/min,flowrateof2.0mL/min.ThenthepurecaulerpinwasobtainedthroughseparatingthecollectedactiveprinciplefromHSCCCbyProteinpurificationapparatus.Therehad0.2317mgofcaulerpininpergramofpinnatefernalgalatthepurityof99.37%.ThestructureofcaulerpinwasauthenticatedbyHPLC-MS,1
6、H-NMRand13C-NMR.Furthermore,theantioxidantactivitiesandantitumoractivitiesofcaulerpinwereinvestigatedaftertheextractionandseparationfromPinnatefernalgal.Fromtheresults,caulerpinshowedmediumantioxidantactivityandthescavengingrate(IC50)for·OH,O2-·andDPPHradicalswere10.8,22.6and
7、23.4mg/mLrespectively.Meanwhile,theantitumoractivitiesofcaulerpinwerealsoapparenttohumanleukemiaHL60cellswithEC50of15.15mg/mL.Hence,alloftheresultsabovesuggestthatcaulerpincanbeusedasaneasilyaccessiblesourceofnaturalantioxidantsandantineoplasticagent.Keywords:HSCCC;Canavaliam
8、aritime;featheryfernfrond;antioxidant;antitumor;IV万方数据目录0前言.........
