The results obtained indicated that PEG-based ATPS were the most effective to separate levodopa from L-phenylalanine while the separation from the other biomolecules was better using IL-based ATPS, in particular those based on [P4444]Cl and [N4444]Cl, with extraction efficiencies of levodopa to the salt-rich phase ranging between 62.7 and 74.0%, and of the remaining biomolecules to polymer/IL-rich phase up to 91.5%.