The mixed monolayer behavior of bilirubin/cholesterol was studied through surface pressure-area （π-A） isotherms on aqueous solutions containing various concentrations of calcium ions. Based on the data of π-A isotherms, the mean area per molecule, collapse pressure, surface compressibility modulus, excess molecular areas, free energy of mixing, and excess free energy of mixing of the monolayers on different subphases were calculated. The results show an expansion in the structure of the mixed monolayer with Ca^2＋ in subphase, and non-ideal mixing of the components at the air/water interface is observed with positive deviation from the additivity rule in the excess molecular areas. The miscibility between the components is weakened with the increase of concentration of Ca^2＋ in subphase. The facts indicate the presence of coordination between Ca^2＋ and the two components. The mixed monolayer, in which the molar ratio of bilirubin to cholesterol is 3：2, is more stable from a thermodynamic point of view on pure water. But the stable 3：2 stoichiometry complex is destroyed with the increase of the concentration of Ca^2＋ in subphase. Otherwise, the mixed monolayers have more thermodynamic stability at lower surface pressure on Ca^2＋ subphase.
Qiong WuYu-feng TangYe-min LiAn-jian XieYu-hua ShenJin-miao ZhuChuan-hao Li
