The three different cultivars of sesame seeds (white, black & brown) procured from local market in Faisalabad, Pakistan were explored for nutritional composition, physico-chemical characteristics, antioxidant activity as well as fatty acid profile. Results showed that seeds contain oil contents ranged from 43.40% to 50.17% with brown seeds having the highest oil contents compared to black & white. Moreover, sesame oil is a rich source of mono and polyunsaturated fatty acids especially oleic acid (56.88%), linoleic acid (50.73%) and palmitic acid (21.67%). These mono and polyunsaturated fatty acids were highest in black sesame seeds compared to white and brown seeds. In addition, total phenols were also highest (30.65%) in black sesame cultivar together with high antioxidant activity (14.83%). Conclusively, black sesame cultivar was found to be the best among all cultivars owing to its high nutritional profile and antioxidant potential.