Lipid Formation in Cryptococcus terricolus. III. Extraction and Purification of Lipids.

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Abstract
Extraction procedures have been developed which release the total lipid content from the yeast C. terricohis without using hydrolytic or high-temperature treatments. The best results were obtained by extracting freeze-dried yeast material at room temperature. Chloroform-methanol (1:1, v/v) was used as solvent, and the material plus solvent were stirred continuously during 3 successive 3-hour extraction periods. The effectivity of the extraction was found to be dependent on the composition of the solvent mixture and on the length and number of the extraction periods. An effective purification of the crude lipids was obtained by a thorough mixing of the extract, dissolved in chloroform-methanol (2:1, v/v), with distilled water, followed by separation by centrifugation. The purified lipids were easily redissolved in the usual lipid solvents. The wash solutions were found to contain only a very small amount of materials of lipid character, varying between 0 and 0.3% of the lipid content of the organism.