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BACKGROUND: The tear film lipid layer is formed from lipids secreted by meibomian glands of the eyelid. After initial analyses of these lipids we concluded that an understanding of the function of the various classes of lipids in a normal lipid layer could only be understood after detailed investigations of both polar and nonpolar lipids of the meibomian gland. METHODS: Meibomian gland secretions were obtained from normals. Lipids were separated by thin layer chromatography and high pressure liquid chromatography, and analyzed by UV absorbance, gas chromatography and mass spectroscopy. RESULTS: Based on our analyses we concluded that the current understanding of lipid layer composition and function were inadequate or misleading. We therefore propose that the more polar lipids function as a structure (with surfactant characteristics) upon which the functional stability of the more nonpolar lipids are dependent. We further suggest that the interrelationships between lipid classes present, length of fatty acids and alcohols, their unsaturation, and hydroxylation are important for maintaining proper thixotropic characteristics of the lipid layer as well as optimal barrier properties. CONCLUSION: The tear film lipid layer is composed of 2 phases: (1) a thin polar phase adjacent to the aqueous-mucin phase and (2) a thick nonpolar phase associated with both the polar phase and the air interface. The structural characteristics of the polar phase and the barrier functions of the nonpolar phase are a direct result of specific compositional parameters.