The axi-symmetric drop-shape analysis-pendant drop technique has been used to measure interfacial tension at the chlorobenzene-water interface in the presence of adsorbed films of dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), DMPC-cholesterol, DPPC-cholesterol, DMPC-cholesterol-dicetyl phosphate (DCP) and DPPC-cholesterol-DCP. A surface-pressure function, pi * = pi lipid-polymer -pi lipid (where pi lipid is the surface pressure of the mono-layer without polymer and pi lipid-polymer is the surface pressure of the lipid mono-layer and adsorbed polymer at equilibrium at the chlorobenzene-water interface) was used to characterize the interaction of eight water-soluble polymers with the lipid films. The equation, delta pi * = pi II*-pi I* (where the subscripts II and I denote the higher and lower lipid composites, respectively) was used to determine the differential effect of cholesterol and DCP on mono-layer characteristics in the presence of 1% w/v polymer. Cholesterol or polymer individually condensed DMPC films and expanded DPPC films. However, composite films of DMPC-cholesterol-DCP and carboxymethylchitin (CM-chitin), poly(acrylic acid) (PAA) or poly(vinyl alcohol) (PVA) were more expanded than DMPC films whereas composite films of DPPC were neither more condensed nor expanded than DPPC films. A polymer impact ratio, P* = pi lipid-polymer/pi lpolymer was calculated and the polymers were ranked in order of their impact on the lipid film. PVA and polysaccharides gave low and high P* values, respectively, corresponding to high and low levels of film interaction, whereas PAA and hydrophobized polysaccharides gave intermediate values, indicating their affinity for and penetration of interfacial films with little disruption of the mono-layer. The results show that measurement of interfacial pressures at the chlorobenzene-water interface might be advantageous for evaluating the action of polymers on biological membranes.