This work presents a comparative analysis of the phenolic composition (UHPLC-PDA-ESI-MS3, HPLC-PDAfingerprint, UV-spectrophotometric methods) and antioxidant activity (DPPH, FRAP) of leaf samples from two vegetation seasons of a medicinal and dietary plant Sorbus domestica growing in its natural habitat (Croatia, C) and cultivated in Poland (P). The samples from both sources were rich in structurally diverse polyphenols (44 analytes; P: 73.4–76.6 and C: 98.3–106.7 mg GAE/g dry leaves) including the dominating flavan-3-ols and flavonoids. The greatest qualitative and quantitative differences were observed for flavonoids (P: 14.3–20.3%; C: 27.5–34.1% of polyphenols) – in the Polish samples flavonoid diglycosides predominated, in the Croatian samples the contents of both monoglycosides and diglycosides were similar. In the case of dry methanolic extracts, despite the higher extraction efficiency obtained for the Croatian samples (32–36% vs 23–24%), the quality of the extracts was comparable, both in terms of the total phenolic content (P: 269.4–280.0; C: 297.6–304.4; mg GAE/g dry extract) and antioxidant activity parameters (DPPH, EC50, μg/mL. P: 10.5–10.9, C: 10.0–10.3; and FRAP, mmol Fe2+/g, P: 6.64–7.13, C: 7.06–7.11). As a result, the study confirmed the influence of environmental conditions on the phenolic profile and antioxidant capacity of S. domestica leaves, as well as showed that despite some differences, plant materials from both Poland and Croatia might be suitable for production of natural health products.