Despite the fact that the wetting properties of multicomponent mixtures including the surface active compounds play a very important role in many practical applications, they are not sufficiently known. Thus, the wettability of polytetrafluoroethylene (PTFE) and poly (methyl methacrylate) (PMMA) by the water + ethanol (ET) solution of rhamnolipid (RL) with Triton X-165 (TX165) mixture was studied. The investigations involved measuring the advancing contact angles of this solution on PTFE and PMMA by varying the concentration of TX165 while maintaining a constant concentration of ET and RL. Additionally, a thermodynamic analysis was conducted to obtain the compositions and concentrations of the ET, RL, and TX165 mixtures at the different interfaces. The composition and concentration of the interface mixed layer were considered using two different approaches to the wetting process. From these considerations, it follows that, depending on the ET concentration, it is possible to form the TX165 + RL layer at the solid–water + ET mixed solvent, as well as the water + ET–air interfaces, but not at the solid–water and water–air ones. This conclusion is in accordance with the Gibbs standard free energy of adsorption of particular components of the studied mixture at the solution–air and solid–solution interfaces.