A_rapid_method_of_total_lipid_extraction_and_purification

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1、CanadianJournalofBiochemistryandPhysiologyIssuedbyTHENATIONALRESEARCHCOUNCILOFCANADAVOI,UME37AUGUST1959NUMBER8ARAPIDMETHODOFTOTALLIPIDEXTRACTIONANDPURIFICATION1AbstractLipiddecompositionstudiesinfrozenfishhaveledtothedevelopmentofasimpleandrapidmethodfortheextractionandpurifica

2、tionoflipidsfrombiologicalmaterials.Theentireprocedurecanbecarriedoutinapproximately10minutes;itisefficient,reproducible,andfreefromdeleteriousmanipulations.'Thewettissueishomogenizedwithamixtureofchloroformandmethanolinsuchproportionsthatamisciblesystemisformedwiththewaterinth

3、etissue.Dilutionwithchloroformandwaterseparatesthehomogenateintotwolayers,thechloroformlayercontainingallthelipidsandthemethanoliclayercontainingallthenon-lipids.Apurifiedlipidextractisobtainedmerelybyisolatingthechloroformlayer.Themethodhasbeenappliedtofishmuscleandmayeasilybe

4、adaptedtousewithothertissues.IntroductionInthecourseofinvestigationsconcerningthedeteriorationoflipidsinfrozenfish,theneedaroseforanefficientandrapidmethodoftotallipidextractionandpurification.Furthermore,duetothehighlyunsaturatednatureoffishlipids,themethodhadtoinvolveonlymild

5、treatmentsoastominimizeoxidativedecompositionandtheproductionofartifacts.Severalexistingmethodswereconsideredbutnonewasentirelysatisfactory.ThemethodsofDambergs(1)andFolchetal.(2)weretootime-consumingforroutineinvestiga-tionsandsincetheformermethodentailedheatingandevaporationi

6、twasconsideredunsuitableforlipidcompositionstudies.TherecentmethodofFolchetal.(3),whichwaspublishedwhilethisstudywasinprogress,wasmorerapidthantheirpreviousmethodbutstillhadthedisadvantageofemployinglargeandinconvenientvolumesofsolvent.ThemethodusedbyDyerandMorton(4)wasrapidbut

7、extractedonlyafractionofthetotallipid.Thepresentpaperdescribesamethodwherebythelipidsofbiologicalmaterialscanbeextractedandpurifiedinasingleoperation.Mixturesofchloroformandmethanolhavehadwideuseaslipidextractantsandexaminationofthechloroform-methanol-waterphasediagram(Fig.1)le

8、dtothefollowinghypothesis.Optimumlipidextractionshould