This research demands to evaluate the oil yield and quality of five enhanced mustard (Brassica spp.) cultivars developed by the Bangladesh Institute of Nuclear Agriculture (BINA). The enhanced varieties are evaluated against traditional local kinds using two distinct extraction methods, cold pressing and hot pressing, as well as during storage. The objective is to determine the influence of extraction method and storage time on oil yield and key quality parameters such as acidity (rancidity), oxidative stability (peroxide value), unsaturation level (iodine value), and proximate composition, seed diameter, density, and refractive index. Mustard varieties used to carry out the study were BINA-4, BINA-9, BINA-10, Sheti, and Deshi. The results revealed that the extraction method significantly influenced both the yield and quality of the oil. Additionally, BINA-9, particularly in the hot-pressed variant, yielded a higher iodine value (10.9 ± 0.026), indicative of a greater degree of unsaturation in the oil. Seed composition analysis reveals significant differences among the seed types, with variations in moisture, crude lipid, ash, carbohydrate, crude protein, and crude fiber content. Oil yield shows a negative correlation (-0.562) with moisture content, as higher moisture potentially reduces oil extraction efficiency. Oil yield has a positive correlation (0.597) with crude lipid content, as higher lipid content usually leads to higher oil yield. This outcome provides critical information on improving mustard oil production techniques, selecting appropriate seed varieties, emphasizing the need of monitoring oil quality throughout storage, and understanding the impact of pressing methods on oil yield and quality.