Lastly, we investigate the e ect of partial charge variability on our calculated AHFEs with a set of 11 FreeSolv molecules which we can also compare back to experiment. We find that the FreeSolv molecules are still subject to maximal partial charge variabilities of up to 0.681 e resulting in ranges of Gs of up to 6.9 ± 0.1 kcal/mol. Despite these molecules being smaller and having fewer unique conformers, we still see significant di erences in partial charge and calculated AHFE. We confirmed these values were due to the partial charges rather than convergence errors by running 2 more repeats of the partial charge sets that led to the minimum and maximum calculated G values. We found the level of calculated G variation remained constant across these additional repeats(Figure ). This suggest that even relatively small molecules are subject to partial charge reproducibility concerns.