Recognition by two-dimensional thin-layer chromatography and densitometric quantification of alkali-labile gangliosides from the brain of different animals

Sandro Sonnino, Riccardo Ghidoni, Vanna Chigorno, Massimo Masserini, Guido Tettamanti

Research output: Contribution to journalArticle

Abstract

A simple method for recognition and quantification of alkali-labile gangliosides is described. The method was worked out using authentic alkali-labile gangliosides in pure form (9-O-Ac-GT1b; 9-O-Ac-GQ1b; lactone form of GD1b) and applied to ganglioside mixtures from the brain of mouse, rat, rabbit, pig, and pigeon. The method consists of two-dimensional thin-layer chromatography on silica gel high-performance thin-layer chromatography plates employing the same solvent, chloroform/methanol/0.2% aqueous CaCl2, 50/40/10, for both runs. Prior to the second run the plate is exposed at room temperature for 5 h to ammonia vapors in order to split alkali-labile linkages. At the end of chromatography alkali-stable gangliosides appear lined along a diagonal starting from the origin; the spots corresponding to alkali-labile gangliosides lie out of the diagonal and can be individually detected and quantified on the basis of their sialic acid content. Up to 15 different spots, corresponding to as many alkali-labile gangliosides, can be recognized by this procedure.

Original languageEnglish
Pages (from-to)104-114
Number of pages11
JournalAnalytical Biochemistry
Volume128
Issue number1
DOIs
Publication statusPublished - 1983

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ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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