Emulsions are utilized in the food, pharmaceutical, and personal care industries to provide specific physicochemical properties and functional attributes. In many applications, it is desirable to use natural ingredients to formulate emulsions to create “label-friendly” products. In this study, the impact of three polysaccharide-based emulsifiers on the formation and stability of oil-in-water emulsions prepared using high-pressure microfluidization were compared: gum arabic, corn fiber gum, and beet pectin. The surface activities of these emulsifiers were characterized using interfacial tension measurements. The influence of emulsifier type, concentration, and homogenization pressure on the efficiency of emulsion formation was examined. The impact of oil type (medium chain triglycerides, corn oil, fish oil, and lemon oil) on the ability of the different emulsifiers to form emulsions was also investigated. The stability of the emulsions was monitored during storage at ambient temperature. Emulsions could be produced using all three polysaccharide-based emulsifiers, with the mean particle diameter decreasing with increasing emulsifier concentration and homogenization pressure. Gum arabic and beet pectin were more effective emulsifiers than corn fiber gum, with a lower amount of emulsifier required and smaller droplets being produced. This effect was attributed to a greater reduction in interfacial tension and stronger adsorption leading to more efficient droplet disruption and less re-coalescence within the homogenizer for gum arabic and beet pectin. Emulsions prepared using corn fiber gum were susceptible to flocculation and coalescence. This study provides valuable information for choosing polysaccharide-based emulsifiers for utilization in food and beverage industries. Display omitted •Emulsions were prepared by high-pressure microfluidization.•Droplet size decreased with emulsifier level and homogenization pressure.•Beet pectin produced smaller droplets than gum arabic and corn fiber gum.•Corn fiber gum emulsions creamed due to large droplets & depletion flocculation.