The genus Potentilla is interesting for the pharmaceutical field due to its valuable medicinal properties, which have been observed in complementary and alternative medicine. In recent years, studies conducted to estimate the biological activity of several of the Potentilla species have shown a wide spectrum of therapeutic properties. In particular, in the present paper, different extracts obtained from the herb P. thuringiaca were analysed for antioxidant and enzyme inhibitory activities. The UHPLC-DAD-MS3 hyphenated techniques reported herein allow for the identification of phytoconstituents. The analyses showed the presence of flavonoids and ellagitannins as major components. Furthermore, the data demonstrated that the analysed extracts revealed a high total antioxidant capacity in the phosphomolybdenum assay. The free radical scavenging activity of the extracts was evaluated using DPPH and ABTS assays. The reducing power activity of P. thuringiaca was also determined by FRAP and CUPRAC assays, as well as metal chelating activity. In addition, the total extracts and the different fractions of P. thuringiaca revealed potent inhibitory activities against α-amylase and α-glucosidase, AChE, tyrosinase and lipase. Surprisingly, no activity against BChE was shown. P. thuringiaca could be a valuable natural source of antioxidants with interesting inhibitory actions against the key enzymes involved in several human diseases, and could represent a valid starting point for the development of new treatment and management strategies, including its use as a food supplement.