The investigation aims to elucidate the bioactive constituents present in waste seeds derived from Nicotiana tobacum L., there by contributing to a more holistic comprehension of the health-related implications associated with tobacco plants beyond conventional consumption concerns. Waste seeds from tobacco plants N. tobacum L. were grown during harvest 2021-2022 at the Tobacco and Tobacco Products Institute (part of Bulgarian Agriculture Academy). Chemical analysis of tobacco waste seeds (TWS) encompassed the examination of primary metabolites including lipids (32.1±1.0%), proteins (29.4±1.4 %), and carbohydrates (27.6±0.5 %). Furthermore, various lipid indices (Atherogenicity, Thrombogenicity, Hypocholesterolemic/hypercholesterolemic, etc.) were computed utilizing the fatty acid composition of the oil extracted from tobacco waste seeds. The results showed that TWS could be utilized with health benefits – seeds are a good resource for n-6 fatty acids (linoleic acid - 71.94±1.05 %) with good atherogenicity and thrombogenicity qualities and natural antioxidants.