The effect of Doxorubicin which is (an anthracycline antibiotic with a broad spectrum of antitumor activity) on the monolayer and bilayer in the form of large Multilamellar Vesicles (MLV's) of Dipalmitoyl phosphatidylcholine (DPPC) were studied by means of monolayer techniques (surface pressure, penetration kinetics, and association constant) and light scattering technique. The monolayer technique showed that addition of DXR to a lipid film composed of (DPPC/CHOL/PEG-PE) at a molar ratio of (100:0:0) produced a less condensed Monolayer. In the (pie-A) curves, DXR induced shift towards larger area/molecule, where the area/molecule was shifted from 61 to 89 A2, and 116 A2 in the presence of 20 and 40 nM DXR, respectively. The three curves collapsed at a pressure pi = 45 mN/m. In penetration kinetics experiment (delta pi-t), the change in pressure with time was 8 and 14 mN/m for a DXR concentration of 20 and 40 nM, respectively, and the increase in surface pressure presented a plateau over a period of 30 min. The measured association constant (K) was found to be 5 x 10(5)/M. In the light scattering experiment, there was a shift of the transition temperature (Tm) of (MLV's) of the same composition of the monolayer towards a smaller value from 40.5 degrees to 34.5 degrees C. Incorporation of CHOL and PEG-PE as DPPC/CHOL/PEG-PE at a molar ratio of (100:20:0), (100:20:4) and (100:20:4) greatly counteracted the effect of DXR and made the lipid membrane more condense and rigid. Moreover, the penetration of DXR into the membrane was greatly reduced. There was a very small shift for the (pi-A) and (delta pi-t) curves, and the association constant of the drug for these different lipid compositions was greatly reduced down to 2.5 x 10(5)/M and the transition temperature (Tm) was increased up to (42.5 degrees C) in the presence of 40 nM DXR. Our results suggest that DXR has a great effect on the phospholipid membrane, and that addition of CHOL or PEG-PE to the phospholipid membrane causes stabilization for the membrane, and reduces the interaction with Doxorubicin.