Biphasic systems such as two-phase partitioning bioreactors (TPPBs) have been used to alleviate biological inhibition by sequestering inhibitory compounds within an immiscible phase. The use of solid polymer beads as this auxiliary phase provides a fully biocompatible alternative to potentially toxic organic solvents. While guidelines exist for the rational selection of the polymer phase, the effect of the aqueous phase composition on molecular sequestration has not been explored in the literature. This work aims to identify aspects of medium composition that influence the partitioning of target molecules into the sequestering phase. Using benzaldehyde as the target molecule and Hytrel G3548L (DuPont) as the polymer phase, pH, temperature, salt and glucose concentrations, as well as ethanol concentrations, were examined for their effects on the partition coefficient. pH and temperature were observed to have no significant effect on benzaldehyde partitioning. Salt and glucose additions increased the partition coefficient by 173% and 30%, respectively, compared with pure reverse osmosis (RO) water, while increasing ethanol concentration was found to decrease the partition coefficient from 44 (+/-1.6) to 1 (+/-0.3). Strategic changes to the aqueous phase can be made to improve affinity of the sequestering phase for target molecules. This provides a simple and cost-effective method to potentially improve TPPB system performance. (C) 2010 Society of Chemical Industry