Ceramides play a major role in structuring and maintaining the water permeability barrier function of the skin, and are widely used in different products for skin care and treatment. This work focused on the isolation of internal wool lipid extracts, enriched in ceramides by supercritical fluid extraction with carbon dioxide (CO2), in an attempt to minimize the environmental impact of the organic solvent extraction process. Selected extraction variables were optimized at two laboratory scales. The lipid composition of the different collected extracts was quantitatively analyzed by thin layer chromatography coupled to an automated flame ionization detector. The extracted wool was physically and chemically evaluated to determine its feasibility for textile purposes. Larger amounts of ceramides were obtained when 10.0 % methanol or ethanol was used as a modifier. Experimental data indicated that a temperature of 60°C resulted in a large yield of ceramides when low pressure (160 atm) and high CO2 volumes (5—7 cell volumes) were applied. Moreover, the temperature and the percentage of the modifier could be reduced to 40°C and to 5.0 % when methanol was used as a modifier, yielding slightly smaller amounts of ceramides. Although slight modifications were observed in some properties of the fibers because of the different extraction conditions, these differences were insignificant.