Sesame, the “queen of oilseeds,” is highly valued for its edible oil, though it contributes very small proportion to the world’s vegetable oil production. Oil content in sesame seeds is close to 50% and protein to 25% of the seed weight. A variety of extraction methods are used to obtain oil from sesame seeds. Among them, mechanical cold press extraction method is highly preferred for oil extraction, as it yields improved oil quality without affecting oil expelling conditions. However, the oil recovery is comparatively lower (~65%) than the chemical methods of oil extraction. Chemical methods are preferred for oil extraction due to higher oil recovery; however, the chemical solvents used had a deleterious impact on the human health and environment which pushed the development of innovative, ecofriendly oil extraction techniques. These advanced techniques employ aqueous extraction, pressurized fluids, phase-partitioning, enzyme-assisted aqueous extraction, etc., which offers numerous advantages such as high extraction yields and quality in short period with minimized solvent utilization. The fatty acid composition of sesame oil depicts that it is high in oleic (40–41%) and linoleic acid (41–43%), which makes it highly nutritive. Also, sesame oil is rich in native antioxidants, viz., lignans (sesamin, sesamolin, sesamol), tocopherols, phytosterols, etc. This chapter also compares the nutritional quality of sesame oil and other vegetable oils. In addition to better oil extraction techniques, there is pressing need to develop efficient techniques for assessing the quality of the extracted oil in order to detect any food fraud committed by mixing low-priced edible oils with high-value oil. Although techniques used in past were reliable, majority of them were destructive assays, hence there is need for improvement in non-destructive methodologies. A range of non-destructive methods that are fast, accurate, and reliable have been devised, which include NIR spectroscopy, FTIR spectroscopy, Raman spectroscopy, hyperspectral imaging (HIS), E-noses, etc. These non-destructive methods can nowadays be implemented through portable analyzers. The information compiled in this chapter may assist oilseed researchers and processors in selecting appropriate method for extracting sesame oil and for assessing its quality.