The archaeological record in the Negro River in central Uruguay Republic is remarkable for its richness, including the presence of a significant number of Paleoindian fishtail points. As part of ongoing research on the earliest human occupations in this region, luminescence dating is applied to develop a terminal Pleistocene-Holocene regional chronology. Eight ages were derived from six samples taken from sedimentary fluvial deposits from two sites near the city of Paso de los Toros. The resulting dates span 11.8 and 1.04 ka corresponding to the Late Pleistocene and the whole Holocene. Because the chronology of the sedimentary sequences spanning these ages is poorly known, the presented results become a significant contribution to the construction of a chronostratigraphy sequence in the area. The results also show how single-grain dating can distinguish different components of mixed assemblages.