In view of the limited information available in the literature concerning leaves as by-products of Prunus armeniaca cultivation, the aim of this work was to identify and characterize their principal polyphenolic constituents by LC-ESI-QTOF-MS/MS and screening in vitro biological potency as antioxidant capacity (ABTS, online ABTS, FRAP, ORAC), antidiabetic (α-amylase, α-glucosidase), anti-obesity (pancreatic lipase), anti-cholinesterase (AChE and BChE), and anti-inflammatory (COX-1 and COX-2) inhibitory activity. Here, ACHE is linked to obesity disorder.