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A special place among vegetable oils was occupied by natural tobacco oil. Natural tobacco oil in folk medicine is used as an antiseptic and antibacterial agent. To determine the possible alternative use of tobacco, the seeds representing Kazakhstan tobacco cultivars, extracted from ground Nicotiana tabacum seeds, were investigated by using the cold press. The quality of the oil was assessed in terms of free fatty acid content. The percentage oil yield was 36.75 ± 0.50%. Tobacco seed oil is highly unsaturated, nonacidic, and would require little purification. The oil can be used for the production of soap, antiwrinkle cream for the face, hair shampoo, shoe cream, and as a high-resolution base oil for perfumes and light industry and can be commercialized due to its high oil yield, for use as anti-inflammatory drugs.