A series of samples for radiocarbon dating were collected in 2005 from the Biblioteca Magliabechiana archaeological excavation area in the center of Florence, Italy, in a project directed by the Dipartimento di Archeologia e Storia delle Arti of the University of Siena, in cooperation with the Soprintendenza per i Beni Architettonici e per il Paesaggio per le province di Firenze e Prato. This area is located near the Uffizi Galleries, close to the Roman town, the Arno River and one of its former tributaries. The area is peculiar for the Florentine urban context because it was free from urban development until the 12th century AD. The exposed stratigraphy showed the presence of several layers composed of natural sediments, partly the result of historical floods. Here, we report a series of 14C measurements on charcoal and seed samples collected on this excavation. 14C dating has been performed in the LABEC laboratory in Florence, on the accelerator mass spectrometry (AMS) beam line of the AMS-IBA 3MV Tandetron accelerator. We also had the opportunity to compare the 14C dates obtained with several series of samples previously collected in nearby archaeological areas. Results were consistent with the data obtained previously and, moreover, offer interesting new aspects to the interpretation of the archaeological findings.