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Silver nanoparticles were obtained via totally green synthesis from silver nitrate using glucose as reducing agent and sucralose as capping agent. Physico-chemical characterization of AgNPs was performed by UV-Visible absorption and transmission electron microscopy (TEM) in order to correlate biological/toxicological responses with AgNPs features. A detailed study of silver nanoparticles (AgNPs) internalization process in human epitheloid cells of cervical cancer (HeLa) was performed versus time. Graphite furnace atomic absorption spectroscopy (GF-AAS) was used in order to assess the silver concentration effectively internalized by the cells. A kinetic model was developed to interpret the results.