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Roots and aerial parts of Cistaceae have been used since ancient times in the Mediterranean cultures for its medicinal properties. In this study, phenolic and tannin content of C. ladanifer and C. populifolius leaves aqueous extracts were determined and their antioxidant and antimicrobial activity were fully studied by several in vitro assays. Their major compounds were identified and quantitated by high-performance liquid chromatography with diode array detection coupled to electrospray ion-trap mass spectrometry. Cytotoxicity on a panel of human cancer cells was also determined. C. populifolius extract was stronger antioxidant than C. ladanifer extract in electron transfer reaction based assays but C. ladanifer extract was more effective to inhibit peroxyl radicals. The major compounds in both extracts were ellagitannins, especially punicalagins derivatives, showing C. populifolius a higher content. C. ladanifer …