Recent research has identified the existence of a freshwater reservoir effect influencing the radiocarbon dating of human skeletal remains from the Dnieper region of Ukraine (Lillie et al. 2009). The current study outlines the evidence for freshwater resource exploitation throughout the period ~10,200–3700 cal BC, and presents the available evidence for the existence of dietary offsets in the 14C dates obtained. We have obtained human skeletal material from 54 Epipaleolithic to Mesolithic period individuals and 267 Neolithic to Eneolithic individuals, from 13 cemeteries, since our research in Ukraine began in 1992. Here, we present the initial results of stable isotope analysis of Eneolithic individuals from the Igren VIII cemetery alongside the Epipaleolithic to Eneolithic samples that have previously been analyzed. When contrasted against the evidence from the prehistoric fauna and fish remains studied, and modern fish species from the Dnieper region, we continue to see variability in diets at the population level, both internally and across cemeteries. We also observed temporal variability in human diets across these chronological periods. The fish samples (both archaeological and modern) show a wide range of isotope ratios for both δ13C and δ15N, which could prove significant when interpreting the dietary sources being exploited. This information directly informs the 14C dating program as an inherent degree of complexity is introduced into the dating of individuals whose diets combine freshwater and terrestrial sources in differing quantities and at differing temporal and/or spatial scales (e.g. Bronk Ramsey et al. 2014).