The separation characteristics of W/O emulsion were investigated under ultrasonic standing wave field in this paper. The effects of the irradiation time, acoustic intensity, frequency, oil viscosity, and oil-water interfacial tension on the separation characteristics of emulsion were studied in details. It was proved that there existed an optimal irradiation time, acoustic intensity and oil -water interfacial tension to achieve the maximum separation efficiency in ultrasonic treatment process. Excessive long irradiation time had little effect on improving the dehydration rate of emulsion but rather increased unnecessary processing time. Excessive high acoustic intensity caused the occurrence of acoustic cavitation phenomenon. The acoustic cavitation would cause the aggregated droplets to be dispersed by oscillating bubbles or even emulsified again by high speed micro jets. The acoustic cavitation threshold is effectively and accurately obtained by a spectral analysis method. The micro acoustic streaming which resulted from the attenuation of acoustic energy occurred with the increase of frequency. It sheared the aggregated droplets, destroyed the stable droplets banding and reduced the separation efficiency of emulsion. The excess surfactant adsorbed on the oil-water interface resulted in the increase of the interfacial film strength, which hindered the coalescence process of droplets and reduced the separation efficiency.