| Summary: | According to recent studies, Norway spruce cones and eastern hemlock cones possess exceptionally high antioxidant capacities; however, the antioxidant efficiency of extractives and their bioactive effects have yet to be investigated in detail. The present article utilizes state-of-the art analytical chemical methodology and chemometric evaluation to reveal cone extractives with the highest antioxidant potential, accounting for possible bioactive effects. Antioxidant properties were determined by the Ferric reducing antioxidant power (FRAP) assay and the Folin–Ciocalteu’s total phenol content (TPC) assay. Structure and relative polyphenol concentrations were determined using liquid chromatography/diode array detection/tandem mass spectrometry. The antibacterial testing was conducted using <i>Staphylococcus aureus</i> and <i>Escherichia coli</i> strains. Using correlation analysis and factor analysis, a scoring evaluation was implemented to determine and compare the antioxidant efficiency of extractives. In Norway spruce, piceatannol-<i>O</i>-hexoside and coumaric acid derivatives were found to be the most powerful antioxidants, while in eastern hemlock, kaempferol glycosides were the most powerful. The antibacterial test did not show any promising antimicrobial activity against <i>E. coli,</i> although a visible inhibitory effect on <i>S. aureus</i> strain was observed.
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