The triesters of phosphatidylcholine as the derivatives of natural phosphatidylcholines are less cytotoxic than the other cationic lipoids, therefore they can be applied in lipofection and in drug delivery. However, a successful and effective use of these compounds requires detailed information of their mechanism of action, which is probably highly complex and multi-stages. However, the first barrier in the way to cell and thus the first side of action of these compounds is the cellular membrane. The aim of this work was to investigate the effect of one cationic lipoid, namely 1-palmitoyl-2-oleoyl-sn-glycero-3-ethylphosphocholine (EPOPC) on model POPC/SM/Chol = 1:1:1 membranes. The experiments were performed on monolayer and bilayer systems and they involved the surface pressure measurements, Brewster angle microscopy studies, dynamic light scattering and zeta potential measurements and the experiments with the surfactant solution and steady-state fluorescence anisotropy of DPH and TMA-DPH. Moreover, to perform the studies systematically also the properties of the binary (POPC/EPOPC, SM/EPOPC, Chol/EPOPC) and ternary (POPC/Chol/EPOPC, SM/Chol/EPOPC) model systems were investigated. The obtained results indicated that even low concentration of EPOPC alters properties and organization of model membranes. Namely, EPOPC makes the interactions in model membrane weaker and increases fluidity and permeability of the lipid system. Finally, based on these data it can be proposed that the mechanism of action of EPOPC in lipofection/drug delivery involves the modifications in membrane organization, which facilitates the incorporation of drug or other material into the cell.