BACKGROUNDXanthohumol is an isopentadienyl flavonoid in hops, which have several pharmacological effects. However, due to the poor bioavailability of xanthohumol, it cannot be widely used.RESULTIn this study, solvent extraction combined with preparative liquid chromatography was used to separate and purify xanthohumol in hop residue. And the purity, yield and recovery of xanthohumol was 983.0 ± 2.1 g kg−1, 921.61 ± 5.65 g kg−1, and 5.41 ± 0.07 g kg−1, respectively. Response surface methodology optimization revealed that 216.75 g kg−1 ethyl oleate, 574.1 g kg−1 polyoxyl‐35 castor oil (EL35) and 209.15 g kg−1 polyethylene glycol 200 (PEG200) produced the xanthohumol nanoemulsion with a loading capacity of 85.40 ± 0.33 g kg−1, mean droplet diameter of 42.35 ± 0.06 nm, and zeta potential of −21.78 ± 0.18 mV.CONCLUSIONXanthohumol nanoemulsion has better relative stability. The relative oral bioavailability of xanthohumol nanoemulsion was increased by 1.76 times. These results provide a theoretical basis for the application of nanoemulsion containing xanthohumol in food and pharmaceutical industry. © 2024 Society of Chemical Industry.