This study was performed to investigate the content of selected phenolic compounds, antioxidant activity and the levels of arbutin and hydroquinone in 25 varieties of highbush blueberry (Vaccinium corymbosum) leaf samples. An analysis of the bioactive components was performed using the HPLC technique and the antioxidant activity was determined via spectrophotometric methods. The content of chlorogenic acid in the analysed leaf extracts ranged from 52.76 mg/g (Spartan variety) to 32.37 mg/g (Nelson variety) and was present in the highest concentration among all the analysed phenolic acids. Particularly large levels of isoquercetin were found in the Aurora, Ivanhoe and Toro varieties (28.40 mg/g, 26.24 mg/g and 21.57 mg/g, respectively). An exceptionally high rutin content (p < 0.05) was found in the Ivanhoe variety (27.19 mg/g) as compared to the other varieties, where it ranged from 2.06 mg/g (Earliblue and Patriot varieties) to 10.55 mg/g (Bluejay variety). The Patriot variety was determined to possess the highest antioxidative activity using the FRAP method (1086.15 μmol Trolox/g d.w.) and based on its DPPH radical scavenging activity (1124.17 μmol Trolox/g d.w.). The total phenolic content (TPC) determined via spectrophotometry ranged from 48.11 mg GAE/g d.w. (Elizabeth variety) to 177.31 GAE/g d.w. (Patriot variety). The arbutin content in the leaves of all tested varieties exceeded 2%, so it can be concluded that they constitute a stable source of arbutin. Three varieties (Bonus, Chanticleer and Herbert) can be considered a potential alternative to bearberry and lingonberry leaves. The hydroquinone content in the analysed extracts was determined to be at a lower level. V. corymbosum leaves can be considered an interesting herbal material for use in traditional herbal medicinal products but not directly for food products and dietary supplements.