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This study was conducted for the comparative analysis of antioxidant activity and untargeted metabolomics of dark- and light-colored sour cherry cultivars grown in Canada. Based on our previous study, we selected four cultivars—'Heimann R', 'Gorsemska', V70142, and 'Montmorency'—to determine the untargeted metabolites and their role in antioxidant activities. A total of 473 metabolites were identified from four sour cherry genotypes using UPLC–ToF–MS. Untargeted metabolomics revealed the dominant chemical groups present in sour cherries. PCA showed that the diversity in sour cherry metabolites was due to the genotype differences indicating iditol, malic acid, chlorobenzene, 2-mercaptobenzothiazole, and pyroglutamic acid as the predominant contributors. The variable importance in the projection (VIP > 1.0) in partial least-squares–discriminant analysis described 20 biomarker metabolites representing the cherry metabolome profiles. A heatmap of Pearson's correlation analysis between the 20 biomarker metabolites and antioxidant activities identified seven antioxidant determinants that displayed the highest correlations with different types of antioxidant activities. TPC and TAC were evaluated using the Folin–Ciocalteu method. The total antioxidant activity was performed using three different assays (ABTS, FRAP, and DPPH). This study of correlating metabolomics and antioxidant activities elucidated that the higher nutritional value and biological functions of sour cherry genotypes can be useful for the development of nutraceutical and functional foods. [ABSTRACT FROM AUTHOR]