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This study divulges a comparative analysis of Berberis tinctoria Lesch. leaf, stem, and fruit towards proximate contents assessment, e.g., moisture, crude-ash, crude-fibre, crude-lipid, crude-protein, nitrogen-free extract, energy followed by ICP-MS analysis which revealed that fruits have a higher amount of essential minerals. The primary metabolites were higher in the fruit extract, whereas more secondary metabolites were observed in the methanolic extracts in the hierarchy of leaf and fruit then stem, respectively. Likewise, a higher amount of antioxidant activities were observed in ethyl acetate and methanolic extracts. The fruit ethyl acetate extract showed a maximum of 95.27% anti-inflammatory activity. The fruit methanolic extract exhibited an antidiabetic activity with IC50 values of 75.5 ± 6.8 μg/mL and 45.4 ± 11.1 μg/mL for α-amylase and α-glucosidase, respectively. A significantly positive correlation was identified between the secondary metabolites and antioxidant activities. The principal component analysis implies ethyl acetate and methanolic extracts as pivotal extracts of B. tinctoria.