In this paper, it is proposed to install two single phase transducers(SPTs) with resonance frequency of 1 MHz in parallel on the inclined glass substrate to conduct microliter oil/water mixed drop separation experiments. Combined with the force of oil/water mixed droplets on the inclined substrate, the two dimensional Navier Stokes equation and the sound flow theory were used to establish the droplet motion mechanics model. The variation relationship between the separation time of the microliter oil/water mixed drop and various influencing factors was analyzed. The theoretical analysis and experimental results show that the factors affecting the separation time mainly include the substrate inclination angle, input peak voltage and oil/water mixing ratio. The oil/water mixed droplets can be efficiently separated by adjusting the value of the influence factors. The research method can reduce the cost of traditional oil/water mixed drop separation, and provide a new idea for separating two incompatible microliter mixed droplets.