Improved separation of isomeric gangliosides by anion-exchange high-performance liquid chromatography.
A new anion-exchange HPLC method for separation of gangliosides has been developed using TMAE (trimethylaminoethyl)-Fractogel as the stationary phase and a gradient system of ammonium acetate in methanol for elution. Chromatography of mouse brain gangliosides resulted in separation according not only to their degree of sialylation into mono-, di-, tri- and tetrasialogangliosides, but also in separation of positional isomers within the four elution domains, e.g. GD1a (IV3Neu5Ac,II3Neu5Ac-GgOse4Cer) and GD1b (II3(Neu5Ac)2-GgOse4Cer) of the disialoganglioside fraction. Furthermore, base-line separation of alpha 2-3 and alpha 2-6 sialylated neolacto-series gangliosides e.g. IV3Neu5Ac-nLcOse4Cer and IV6Neu5Ac-nLcOse4Cer, isolated from human granulocytes, was achieved and pure fractions of each ganglioside were obtained. Thus, the high-resolution power of the strong anion-exchanger TMAE-Fractogel allowed the isolation of pure gangliosides on a preparative scale by one-step column chromatography.
Müthing
J
J
Institute for Cell Culture Technology, Faculty of Technical Sciences, University of Bielefeld, Germany.
Unland
F
F
D000085:Acetates; D000818:Animals; D000838:Anions; D001923:Brain Chemistry; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange; D005732:Gangliosides / Q000302:isolation & purification*; D006098:Granulocytes / Q000737:chemistry*; D006801:Humans; D007202:Indicators and Reagents; D000432:Methanol; D051379:Mice; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid; D011108:Polymers; D000644:Quaternary Ammonium Compounds; D012794:Sialic Acids / Q000378:metabolism
D000085:Acetates; D000838:Anions; D005732:Gangliosides; D007202:Indicators and Reagents; D011108:Polymers; D000644:Quaternary Ammonium Compounds; D012794:Sialic Acids; C090635:trimethylaminoethyl-fractogel; D019158:N-Acetylneuraminic Acid; C018824:ammonium acetate; D000432:Methanol