A Variant for Isolation of Serum Gangliosides

The serum gangliosides are of great diagnostic importance. A step-by-step procedure for their isolation from serum is described and it includes the following stages: a) dehydration of the sample; b) total lipids extraction; c) non-polar lipids removal by preparative TLC; d) elimination of the blood sugar by Sep-Pak technique; e) TLC of the ganglioside fractions. Azeotropic distillation of the mixture of serum water/n-propanol was used for sample dehydration. Triple extraction was carried out: with cyclohexane (I); chloroform : methanol = 1 : 1 (v/v) (II) and chloroform : methanol = 1 : 2 (v/v) (III). The extracts were combined and the non-polar lipids were removed. The eluate composition was checked out by HPTLC with a modified mobile phase: chloroform:methanol:0.1 M sodium lactate = 55 : 40 : 10 (v/v/v). Comparative data for the R f values of the ganglioside fractions as well as recovery data are presented. This method is also applicable for milk samples.