Poly and perfluoroalkyl substances (PFAS) are a large group of emerging organic pollutants that can persist in the environment and bioaccumulate in biota. They are found in complex mixtures, and although the exact number of PFAS is unknown, it has been estimated to be in the thousands. The objective of this study was two-fold. First, we examined the cytotoxicity of PFAS singly and in binary mixtures using an amphibian fibroblast cell line. Second, we used this experimental data to develop quantitative structure-activity relationship (QSAR) models for single and binary mixtures. We tested the cytotoxicity of four common PFAS: perfluorooctane sulfonate (PFOS); perfluorooctanoic acid (PFOA); perfluorohexane sulfonate (PFHxS); and perfluorohexanoic acid (PFHxA). PFOS was the most toxic and PFHxA the least cytotoxic. Binary mixtures allowed for the construction of isobolograms to test for additivity, synergism, or antagonism. Using this data, QSAR modeling was used for predicting the toxicity of 24 single and 1380 binary mixtures (theoretically generated). Overall, our experimental and modeling results showed that mixtures were approximately additive, with the exception of PFOS and PFOA, which were found to be weakly synergistic. This data shows that certain mixtures of PFAS may have increased toxicity potential above what the simple sum of PFAS concentrations would suggest. More studies are needed that test the toxicity of PFAS mixtures.