This study aimed to develop and optimize the nanoemulsion formulation of asiaticoside (AS) using Box-Behnken design. Nanoemulsion optimization was used to obtain the optimum formula for ideal characteristics. Nanoemulsions were formulated using a combination of high-speed homogenization and ultrasonication. The independent variables were virgin coconut oil (VCO) as the oil phase, sucrose ester with HLB 3 (SE) as a surfactant and sodium caseinate (SC) as a co-surfactant. Particle size, polydispersity index (PDI) and zeta potential were the dependent variables used as responses to determine the optimum formula. The best combination of the composition factors that gave the optimum response was found to be VCO (13.55%, w/w), SE (2.5%, w/w) and SC (2%, w/w). The obtained nanoemulsion optimum formula was evaluated according to polydispersity index, particle size and zeta potential with the results obtained at 0.146 ± 0.002, 262 ± 1.000 nm and-62.9 ± 1.365 mV, respectively. The developed nanoemulsion formulation using the Box-Behnken design could be used as the optimal formulation of asiaticoside and to determine the effects of the formulation variables. © 2024, Romanian Society for Pharmaceutical Sciences. All rights reserved.