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        Desalting Acidic Glycosphingolipids

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        <center> <h2> <font>Desalting Acidic Glycosphingolipids</font></h2> </center> Contributor: Suprya Jayadev
        Date: November 10, 1993


        1) Prepare a C-18 reverse phase prep-sep cartridge by washing with:

        a) 10 ml of methanol

        b) 20 ml of chloroform-methanol (2:1)

        c) 10 ml of methanol

        d) 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 methanol

        b) 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).


         

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