Ceramides extracted from wool fibers have proved to be beneficial in pharmaceutical and cosmetic formulations in the treatment and care of skin and human hair. This work sought to obtain internal wool lipid extract enriched in ceramides at pilot plant level without causing significant damage to the wool fiber. To this end, two extraction methodologies were performed using two solvents — acetone and methanol. Analyses of wool extracts were performed by thin layer chromatography coupled to an automated ionization detector, and chemical and mechanical evaluations of extracted wool were carried out. Larger amounts of ceramides were obtained when wool fibers were extracted with methanol than with acetone with some modifications of the fiber properties. This lipid extract could be a by-product with a high added value for the wool industry.