Contributor:SupryaJayadevDate:November10,19931)PrepareaC-18reversephaseprep-sepcartridgebywashingwith:a)10mlofmethanolb)20mlofchloroform-methanol(2:1)c)10mlofmethanold)10mlofmethanol-1.6Msodiumacetate(1:1)2)Applysamplein2mlofmethanol-1.6Msodiumacetate(1:1).3)Collectwashthroughandapplytwicemoretothecartridge.4)Washcartridgewith40mlofwater.--->Thisactuallywashesawaythesaltfromtheglycosphingolipids.5)Eluteacidglycosphingolipidswith:a)2mlofmethanolb)10mlofchloroform-methanol(1:1)6)Poolthetwoeluatesanddrylipidsdown.7)ResUSPendinaminimalvolumeofchloroform-methanol(1:1)andspotonTLC.8)DevelopTLCinchloroform-methanol-0.22%calciumchloride(60:35:8).