Improved thin-layer chromatographic separation of gangliosides by automated multiple development.
Automated multiple development chromatography has been utilized to enhance separation of gangliosides on silica-gel precoated high-performance TLC plates. Three-fold chromatography of a complex mixture of neolacto-series monosialogangliosides in the solvent chloroform-methanol-water (120:85:14, v/v, 2 mM CaCl2) resulted in a ca. three-fold increase in separation distance of e.g. alpha 2-3 and alpha 2-6 sialylated ganglioside isomers compared to conventional single chromatography in the standard solvent chloroform-methanol-water (120:85:20, v/v, 2 mM CaCl2). An extremely heterogenous murine disialoganglioside mixture was developed three times in chloroform-methanol-water (120:85:16, v/v, 2 mM CaCl2) leading to a more than two-fold increase in separation distance. Chloroform-methanol-water (120:85:22, v/v, 2 mM CaCl2) was the solvent of choice for multiple chromatography of ganglio-series polysialogangliosides from embryonic chicken brain.
Institut für Zellkulturtechnik, Universität Bielefeld, Germany.
J
J
Müthing
D000818:Animals; D001322:Autoanalysis; D001921:Brain / Q000196:embryology; D001923:Brain Chemistry; D002240:Carbohydrate Sequence; D002642:Chick Embryo; D002725:Chloroform; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer / Q000379:methods*; D005732:Gangliosides / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007962:Leukocytes / Q000737:chemistry; D008223:Lymphoma / Q000737:chemistry; D000432:Methanol; D051379:Mice; D008969:Molecular Sequence Data; D014407:Tumor Cells, Cultured; D014867:Water
D005732:Gangliosides; C025447:sialogangliosides; D014867:Water; D002725:Chloroform; D000432:Methanol