Acetonitrile as adjuvant to tune polyethylene glycol + K3PO4 aqueous two-phase systems and its effect on phenolic compounds partition

Abstract

Aqueous two-phase systems (ATPS) have been proposed as platforms for the purification of biomolecules, and the application of adjuvants to tune the properties of ATPS phases and their ability to extract and separate biological products investigated. This work addresses, for the first time, the use of acetonitrile (ACN) as adjuvant in ATPS composed of polyethylene glycol (PEG) of different molecular weights (from 400 to 20,000 g·mol−1) and potassium phosphate. The effect of ACN concentration (at 0.25, 2.5, 5 and 7.5 wt%) in the liquid-liquid equilibrium is here studied by measuring the phase diagrams, the composition of the phases in equilibrium and their Kamlet-Taft parameters. The results obtained demonstrate that the ACN presence increases the biphasic region of PEG-K3PO4 ATPS and its distribution between the coexisting phases is dependent on its concentration and ATPS composition. The difference in the dipolarity/polarizability of the coexisting phases is significantly affected by the presence of ACN in the ternary mixture. The partition of two phenolic compounds – protocatechuic acid and gallic acid – was studied, showing that ACN improves the partition of these molecules to the salt-rich phase, and that the system selectivity can be significantly improved by changing the concentration of the phase forming compounds.