The surface behavior of a series of cholesteryl esters was studied in mixtures with a model phospholipid, 1-palmitoyl-2-oleoyl phosphatidylcholine. The cholesteryl esters were representative of the predominant forms occurring naturally and qualitative similarities in their phase behavior permits generalization of their surface properties. Quantitative differences, however, show that the availability of cholesteryl esters in all surface states is dependent on the structure of the acyl moiety. All except cholesteryl stearate were surface-active and formed preferred packing arrays, i.e., complexes, with the lecithin at compositions grouped around cholesteryl ester mol fractions of 0.015. Exceptions were cholesteryl arachidonate and docosahexaenoate, with complex stoichiometries of 0.034 and 0.032, respectively. Lecithin had the same apparent area in all complexes, 56.5 +/- 1.04 A2, which was larger than that of uncomplexed lecithin, 53.3 +/- 0.7 A2. This implies that the conformation or orientation of the two polyunsaturated species in complexes is markedly different than the others studied. The areas and hydrations of all uncomplexed cholesteryl esters were similar. Because mixing of complex with uncomplexed cholesteryl ester deviated positively from ideality, the apparent molecular areas of the uncomplexed cholesteryl esters ranged from 161 +/- 22 (complex-rich) to 107 +/- 15 A2 (cholesteryl ester-rich). The similarity of the monolayer phase complex stoichiometries and the bilayer miscibilities of cholesteryl oleate suggests a correspondence between states. If so, the availability of cholesteryl arachidonate or docosahexaenoate in bilayers should be approximately twice that of other naturally occurring cholesteryl esters.