The present study was carried out to fabricate the food grade Vitamin E acetate nanoemulsion using edible mustard oil and to evaluate its improved bioactivities. A food-grade Vitamin E acetate nanoemulsion was fabricated using the edible mustard oil and surfactant Tween-80. Flocculation was not observed for 15 days. The nanoemulsion was characterized for droplet morphology and size distribution using atomic force microscope and zetasizer, respectively. We observe a stable nanoemulsion of spherical morphology and a size distribution of 86.45 ± 3.61 nm. Further, the high-performance liquid chromatography method was used to determine the Vitamin E acetate concentration and encapsulation efficiency for the stable nanoemulsion. These nanoemulsions showed improved bioactivity, antioxidant, and antimicrobial activity and could be potentially used to increase the shelf life of fruit juice. Copyright © 2015 Taylor & Francis Group, LLC.