Detection of gangliosides with the fluorochrome NBD dihexadecylamine and its application for preparative high performance thin layer chromatography.
A new method for the detection of gangliosides based on the lipophilic fluorescence agent 4-(N,N-dihexadecyl)amino-7-nitrobenz-2-oxa-1,3-diazole (NBD dihexadecylamine) and its application for preparative high performance thin layer chromatography is described. Brain gangliosides were chromatographed on silica gel coated thin layer plates and located with non-destructive fluorochrome under longwave ultraviolet light. The fluorescent zones were scraped off and the gangliosides were extracted with a mixture of chloroform/methanol/water (30/60/8; v/v/v). The gangliosides were separated from uncharged NBD dihexadecylamine by anion exchange chromatography and impurities were removed by Iatrobeads chromatography. The method described offers a simple and successful preparative thin layer chromatographic strategy to obtain pure gangliosides in microgram and milligram quantities.
Müthing
J
J
Institut für Zellkulturtechnik, Universität Bielefeld, Germany.
Unland
F
F
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives*; D000818:Animals; D001923:Brain Chemistry; D002725:Chloroform; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes; D005732:Gangliosides / Q000032:analysis*; D006801:Humans; D000432:Methanol; D051379:Mice; D008808:Mice, Inbred CBA
D005456:Fluorescent Dyes; D005732:Gangliosides; C078648:4-(N,N-dihexadecyl)amino-7-nitrobenz-2-oxa-1,3-diazole; D002725:Chloroform; D009327:4-Chloro-7-nitrobenzofurazan; D000432:Methanol