The behavior of hesperidin methylchalcone (HMC) at the oil/water interface was examined through experimental and molecular simulation methods, and a nano-emulsions based on HMC was subsequently fabricated. The findings indicated that HMC spontaneously aggregated at the oil-water interface, leading to a reduction in interfacial tension and an increase in interfacial thickness. Furthermore, its glycoside and benzene ring showed tendencies to interact with water and medium-chain triglyceride, respectively. The HMC addition amount (w), homogenization times (n) and homogenization pressure (p) significantly influenced the formation of the nano-emulsions. The nano-emulsion with an oil-droplet size of 277.26 ± 13.62 nm was obtained at w = 1.0 %, p = 200 bar, and n = 6. When compared to the Tween 20 nano-emulsion, the HMC nano-emulsion demonstrated superior storage stability, antioxidant activity, and lutein bioaccessibility. It could achieve the slow release of HMC. These findings not only broaden the application range of HMC but also contribute to the advancement of functional nano-emulsions.