Inclusion complex formation between β-cyclodextrin (β-CD) and suitable guest molecules such as cholesterol (Ch) has regularly been exploited to design self-assembled structures. In the present study an effective nanoemulsion medium (lecithin/Tween 80/ethyl oleate/water) was selected for solubilizing and stabilizing Ch and Ch/β-CD inclusion complex. Phase solubility, spectroscopic, thermodynamic, Z-average diameter and morphological analyses were conducted. Phase solubility data analysis demonstrated an increase in Ch solubility at low β-CD concentrations (0.01-0.35 mmol L(-1) ). Transmission electron microscopy and Z-average diameter data indicated the spherical nature of the droplets and confirmed the formation of nanoemulsions with an average size of less than 50 nm. The negative value of ΔG obtained during analysis further indicated that the binding was spontaneous in nature. Primarily, this research demonstrates the use of nanoemulsions as a medium in food matrices, instead of water, for hosting Ch in β-CD.