In this investigation, the chemical and biological profiles of three Turkish Crataegus species (C. orientalis, C. szovitsii and C. tanacetifolia) were studied in order to provide the first comprehensive characterization and their health-promoting potential. In this respect, polyphenolic profiles were evaluated using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS). The in vitro antioxidant activities of the Crataegus samples were evaluated by using free radical scavenging, phosphomolybdenum, ferrous ion chelating, and reducing power assays. The inhibitory activities against α-glucosidase, amylase, cholinesterases (acetylcholinesterase (AChE) and butyrylcholinesterase (BChE)), and tyrosinase were also established.The untargeted metabolomic approach highlighted the effect of both extraction types and species on the phenolic profiles of different Crateagus tissues (i.e., leaves and twigs). The methanolic extracts showed the maximum radical scavenging and reducing activity in all test systems, while for ferrous ion chelating assays, the decocted and infused extracts showed the highest activity. Only the methanolic extracts were effective against AChE and BChE. The tested extract showed remarkable inhibitory effects against tyrosinase and α-glucosidase, while all the extracts exhibited modest inhibition against α-amylase. Overall, the twig extracts of the three studied species showed superior antioxidant and enzyme inhibitory activities.On the basis of these results, the three Crataegus species can be classified as potent bio-resources for high value phytochemicals, which warrant further investigations for developing novel nutraceuticals. This article is protected by copyright. All rights reserved.