The outermost layer of the skin, the stratum corneum, consists of non-viable keratin-filled cells, or corneocytes, embedded in a matrix of lipids. The boundary of the cells consists of cross-linked proteins with covalently bound lipids on the outer surface. The spaces between cells are filled with a mixture of ceramides, cholesterol and fatty acids. The stratum corneum provides a protective barrier against water loss through the skin and limits the penetration of potentially harmful substances from the environment. Among the covalently bound lipids on the corneocyte surface are omega-hydroxyacylsphingosine and omega-hydroxyacyl-6-hydroxysphingosine. The previously suspected presence of omega-hydroxyacylphytosphingosine is confirmed in this report through its specific isolation and characterization based on chromatographic behaviour and proton magnetic resonance spectral data.