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Omega-3 fatty acids, which consist of alpha-linolenic acid (ALA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA), are a type of polyunsaturated fatty acids that are crucial for enhancing human health. These three omega-3s are regarded as vital dietary nutrients because it cannot synthesize them on its own. Therefore, they must be obtained through dietary sources. On the other hands, there are concerns regarding the quality and quantity of omega-3 in dietary sources and supplements. In this study, 1H NMR spectroscopy and multivariate analysis were harnessed for non-destructive profiling of fatty acids in commercially available omega-3 supplements and plant-based oils. Results disclosed substantial disparities in omega-3 content from 8 to over 50 %, with some revealing unexpected adulteration. Notably, in one fish oil sample DHA could not be detected indicating the possibility of adulteration. Additionally, the research delineated the fatty acid composition of plant-based oils, emphasizing variations in alpha-linolenic acid (ALA) and linoleic acid (LA) content among flaxseed, chia seed, and walnut oils. Chia seeds emerged as a rich source of ALA (57–65 %mol), while walnuts contained considerable LA (44–53 % mol). The findings emphasize the power of metabolomics as a rapid and affordable tool for finding omega-3 fatty acids content and adulteration in commercial dietary products. • 1H NMR metabolomics is a potent tool for identifying adulteration in ω-3 sources. • Analytical methods based on 1H NMR spectroscopy are rapid and reliable. • Chemometrics equations used to quantify the different classes of ω-3 fatty acids. • Chia seeds had the highest amount of alpha-linolenic acid than flaxseeds and walnuts. • Based on the findings, pharmaceutical industries can choose optimal source of omega-3 fatty acids. [ABSTRACT FROM AUTHOR]