Plants from the genus Potentilla have been traditionally used for therapeutic purposes in Turkey. Methanol root extracts from four selected Potentilla species (P. anatolica, P. argentea, P. recta, and P. reptans) were probed for their chemical composition, antioxidant properties (DPPH, ABTS, FRAP, CUPRAC, phosphomolybdenum, and metal chelating activity), and enzyme (acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, α-glucosidase, and tyrosinase) inhibition effects. In addition, the total phenolic, flavonoid, saponin, and triterpenoid content of the extracts were determined. Ultra-high performance liquid chromatography (UHPLC)-electrospray ionization (ESI)-tandem mass spectrometry (MS/MS) analysis revealed that the main phenolic components in extracts were coumarin, hesperidin, p-coumaric acid, gallic acid, and rutin. P. recta and P. reptans showed the highest total phenolic, saponin, and triterpenoid content. The root extracts showed significant antioxidant properties and depicted significant inhibitory effects against AChE, BChE, α-amylase, α-glucosidase, and tyrosinase. In conclusion, root extracts of Potentilla may be considered as new natural antioxidants and key enzyme inhibitors that can be exploited as bioproducts for pharmaceutical industries.