Local anesthetics are substances that reversibly block the nerve-impulse conduction, alleviating pain without loss of consciousness. Benzocaine, a poorly soluble local anesthetic, is an ester of para-aminobenzoic acid. Several strategies of formulations can be used to improve bioavailability and decrease adverse effects of benzocaine. In this study nanostructured lipid carriers (NLC) were employed. These lipid-based drug delivery carriers have a lipid core composed of a blend of solid and liquid lipids, and a shelf of non-ionic surfactant. The main aim of this work was to optimize benzocaine-loaded NLC and to investigate structural changes in these nanoparticles, under different temperatures. The ratio of excipients (cetyl palmitate, Capmul?? PG-8 NF and Pluronic??F68) and benzocaine in the NLC was optimized using a 2 3 factorial design with respect to the following parameters: particle size, polydispersity index (PDI) and zeta potentials. The interactions between the factors were found relevant to determine particle size and PDI. Using desirability function, the best formulation conditions were found. Structural changes in optimized NLC were observed with Small-Angle Neutron Scattering (SANS) and Raman imaging, in samples at 27, 37 and 40?? C. SANS pointed the formation of lamellar structures inside the NLC, which interlamellar distances increase at higher temperature. Raman imaging showed that the incorporation of P68 and benzocaine in-between the lipids increased at higher temperatures, explaining the changes in Q values (SANS). This work shows how different scattering techniques can provide complementary information and be used together to characterize and understand the physical, chemical, and structural changes on the organization of pharmaceutical carriers in drug delivery system.