HypothesisAsphaltenes are primary stabilizers in water-in-oil (W/O) emulsions that cause corrosion and fouling issues. In oilsands industry, oil/water separation processes are generally conducted at high temperatures. A high temperature is expected to impact the interactions between asphaltenes and emulsion breakers (EBs), consequently influencing demulsification performance. ExperimentsThe adsorption and interactions of asphaltenes and a PEO-PPO type EB (Pluronic F68) at the oil–water interface were investigated at various temperatures, using tensiometer, quartz crystal microbalance with energy dissipation (QCM-D), and atomic force microscopy (AFM). The effect of temperature on EB’s demulsification performance was explored through bottle tests. Additionally, demulsification mechanisms were studied using direct force measurements with the droplet probe AFM technique. FindingsDynamic interfacial tension and QCM-D results demonstrate that the PEO-PPO type EB exhibits higher interface activity than asphaltenes and can disrupt rigid asphaltene films at the oil–water interface. Elevated temperatures accelerate the displacement of adsorbed asphaltenes by EB molecules, leading to sparse interfacial films and rapid droplet coalescence (supported by AFM results). This work provides valuable insights into interfacial interactions between asphaltenes and EB under different temperatures, enhancing the understanding of demulsification mechanisms and offering useful implications for the development of efficient EBs to enhance oil/water separation performance.