A novel technique to investigate the interactions between bitumen and clays in an aqueous solution from the measurement of zeta potential distributions is described. For a single component suspension (i.e., clay or bitumen), a single modal zeta potential distribution was obtained under a given solution condition. In the case of a two-component (i.e., bitumen and clay) mixture system, the measured zeta potential distribution showed either one or two distribution peaks, depending on the chemical condition of the suspension and the type/amount of clays present. In the absence of added calcium ions, a mixture of bitumen emulsion and clay suspension exhibited two distinct zeta potential distribution peaks, corresponding to the peaks measured individually for the bitumen and clays, respectively. With the addition of 1 mM calcium ions, however, only one zeta potential distribution peak was obtained for the mixture of bitumen emulsion and montmorillonite clay suspension. Depending on the montmorillonite clay to bitumen ratio, the peak position in this case shifted toward the value for montmorillonite clay suspension alone. For kaolinite, the addition of 1 mM calcium ions did not cause a substantial change in the bimodal zeta potential distribution. The results suggest qualitatively a stronger interaction of bitumen with montmorillonite clay than with kaolinite clay, when calcium ions were present. The slime coating of montmorillonite clay on bitumen droplets in the presence of 1 mM calcium was validated. The conclusions obtained from this study further justified our mechanistic hypothesis for the observed depression of bitumen flotation by montmorillonite but not by kaolinite clay addition when calcium ions were added. This study demonstrated that zeta potential distribution measurement could be a powerful tool to study slime coating phenomena in a complex colloidal system.