Facile Methods for Isolation and Determination of Gangliosides in a Small Scale: Age-Related Changes of Gangliosides in Mouse Brain Synaptic Plasma Membranes

A quantitative isolation method for gangliosides from small sizes of samples has been developed. Total lipids were separated into gangliosides and other lipids using Phenyl Sepharose column chromatography. Gangliosides were recovered in a yield of 99%. Determination of gangliosides was performed by gas chromatography-mass spectrometry using a selected ion-monitoring technique. These combined methods can provide quantitative isolation and determination of gangliosides from as little as 10 mg fresh brain tissues or 0.5 mg protein of membrane fractions. The present methods were successfully applied to the analysis of gangliosides from mouse brain synaptic plasma membranes to reveal that the ganglioside contents and composition remain constant from adult to senescence.