Abstract: The demulsification of the emulsion formed during aqueous enzymatic extraction of camellia oleifera seed oil by alkaline was studied.Demulsification rate was taken as index.Based on single- factor experiments,the main factors were optimized with response surface methodology( RSM). By employing RSM design programmed,the optimal parameters were generated as follows: p H = 9.0,temperature 60℃,time 45 min and demulsification rate was90.15%.Fluorescence microscopy technology was applied to observe the change of oil droplets before and after alkaline demulsificatication,respectively. The comparision found that alkaline demulsificatication oil droplets aggregated significantly,grease easier to release.