Nondestructive Detection of Gangliosides with Lipophilic Fluorochromes and Their Employment for Preparative High-Performance Thin-Layer Chromatography

✍ Scribed by J. Muthing; D. Heitmann

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 382 KB
• Volume: 208
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1993.1016

✦ Synopsis

A simple and effective procedure for the isolation and purification of gangliosides by preparative thin-layer chromatography is described. The method is based on nondestructive visualization of gangliosides on silica gel-precoated thin-layer chromatography plates by staining with uncharged lipophilic fluorochromes. Fluorescent dyes were added in low concentrations into the mobile phase (0.002%, w/v) without any interference of the ganglioside separation. After uv localization, the fluorescent zones were scraped off the plate and the silica gel was extracted with chloroform/methanol/water (30/60/8). In the following step fluorochromes were removed from gangliosides containing crude extracts by anion-exchange chromatography on DEAE-Sepharose. After desalting, impurities were removed by Iatrobeads chromatography. The method described offers an easy to handle and successful preparative thin-layer chromatography strategy to obtain pure gangliosides in microgram and miligram quantities.