Desalting Acidic Glycosphingolipids

 Desalting Acidic GlycosphingolipidsContributor: Suprya JayadevDate: November 10, 19931) Prepare a C-18 reverse phase prep-sep cartridge by washing with:a) 10 ml of methanolb) 20 ml of chloroform-methanol (2:1)c) 10 ml of methanold) 10 ml of methanol-1.6 M sodium acetate (1:1)2) Apply sample in 2 ml of methanol-1.6 M sodium acetate (1:1).3) Collect wash through and apply twice more to the cartridge.4) Wash cartridge with 40 ml of water.---> This actually washes away the salt from the glycosphingolipids.5) Elute acid glycosphingolipids with:a) 2 ml of methanolb) 10 ml of chloroform-methanol (1:1)6) Pool the two eluates and dry lipids down.7) Resuspend in a minimal volume of chloroform-methanol (1:1) and spot on TLC.8) Develop TLC in chloroform-methanol-0.22% calcium chloride (60:35:8).