Abstract
An improved method for isolating cerebrosides from natural sources is described. The method is particularly suited to large scale work and can be adapted to the isolation of sphingolipids that are less polar than the gangliosides. It is based on the use of sodium sulfate to absorb the water from chloroform-methanol tissue extracts, the use of triiodide to cleave the ether linkage of plasmalogens, and the use of alkaline methanolysis to cleave the ester linkages of the glycerolipids. The final separation of the lipids is done with a silica gel column.
Highlights
T h e alkenyl ether linkage was cleaved by reaction with iodine [1, 2]
The water in the initial lipid extract, arising from the original tissue, was absorbed by anhydrous sodium sulfate, which is readily removed by filtration
Since there was little that was unconventional about our procedure, it may be presumed that the cerebroside purity is comparable to that of other preparations
Summary
Scale work led to the following approaches. T h e alkenyl ether linkage was cleaved by reaction with iodine [1, 2]. The water in the initial lipid extract, arising from the original tissue, was absorbed by anhydrous sodium sulfate, which is readily removed by filtration. Conditions of extraction were chosen to give reasonably complete extraction and fast filtration with a relatively small volume of solvent. Reexamination of the load capacity of silica gel led to the finding that a satisfactory separation could be achieved with a smaller column and with less solvent than previously believed necessary. The procedure described below was designed for 5 0 0 g batches of brain or Gaucher spleen, but its advantages apply to smaller samples. Suggestions are offered for modifications of the method for laboratories lacking some of the equipment used here
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