In Annurca apples, a southern Italian variety, polyphenols were studied. The phenolic composition of Annurca apple peel was determined by HPLC coupled with electrospray negative ionisation multistage ion trap mass spectrometry (HPLC/ESI–ITMS n ). In addition, the antiproliferative and pro-apoptotic activities of peel extracts enriched in polyphenols and prepared from Annurca (APE), Red Delicious (RDPE) and Golden Delicious (GPE) varieties were evaluated on an HL-60 cell line. APE exhibited the highest total phenol content among the three apple cultivars tested, as determined by Folin–Ciocalteau’s procedure and HPLC–UV analysis. APE also showed higher amounts of chlorogenic acid, procyanidins, flavonols, dihydrochalcones and cyanidin-3- O-galactoside than GPE or RDPE. All three apple extracts were able to significantly decrease cell viability, from 50% to 80%, with APE appearing the most effective, while GPE was the least cytotoxic among the three samples. Similarly, APE and RDPE induced a significant increase of caspase-3 activity, whose activation represents a hallmark of apoptosis.