Mafenide acetate is a commonly known antimicrobial agent for wound infection. Permeability of mafenide acetate through eschar is very high and it may lead to systemic toxicity after topical application. We wish to investigate whether topical use of mafenide acetate – including vesicles could result in drug trapping in rat skin, in comparison to mafenide acetate aqueous solution. In this study, liposomes were prepared with two techniques: Solvent evaporation and Microencapsulation vesicular (MCV). We applied full factorial design for experimental design and data analysis. Drug/lipid ratio, hydration time, aqueous phase volume and homogenizer rpm were considered as independent variable, on the other hand, liposome size, drug loading, stability, drug release and skin permeability parameters as responses. The results demonstrate that liposome were multilamellar and multivesicular. Particle size and drug loading percentage of MCV liposome indicated burst sustained release profile. Burst effect in solvent evaporation liposome was more than MCV liposome. In conclusion, solvent evaporation liposome improved mafenide acetate partitioning through rat skin and decrease diffusion coefficient with increase particle size of liposome.