Effects of sodium chloride and sodium perchlorate on properties and partition behavior of solutes in aqueous dextran-polyethylene glycol and polyethylene glycol-sodium sulfate two-phase systems

Abstract

Effects of two salt additives, NaCl and NaClO₄, at the fixed concentrations of 0.215 M on the properties of aqueous two-phase systems (ATPSs) formed by dextran (Dex) and polyethylene glycol (PEG), and the effects of NaClO₄ at the same concentration on the properties of ATPS formed by PEG and Na₂SO₄ were examined. The effects of these salt additives on partitioning of 12 small organic compounds and five proteins in the above ATPSs were studied. In each system with a given salt additive, 0.5 M sorbitol, 0.5 M sucrose, and 0.5 M and 1.5 M trimethylamine N-oxide (TMAO) were also used as additives. The results obtained were compared with those reported previously for the Dex-PEG ATPS without salt additives and PEG-Na₂SO₄ ATPS without salt additives and in the presence of 0.215 M NaCl. It is shown that the differences between the solvent properties of the phases in the systems formed by polymer and salt exceed those observed in the systems formed by two polymers. The three most significant solvent features of the systems are hydrophobic and electrostatic properties and hydrogen bonding donor acidity of the solvent media. Osmolyte additives were found to have a significant effect on the differences between the electrostatic properties of the phases. Analysis of the partition coefficients of 12 organic compounds and five proteins showed that the osmolyte additives may affect the partition behavior of compounds in a compound-specific manner. The relative contributions of different types of interactions of a given compound with aqueous media change in the presence of salt and osmolyte additives. Analysis of the variability ranges of partition coefficient, K, in the systems studied showed that for small organic compounds, the ranges of K-values observed in the PEG-Na₂SO₄ ATPSs exceed those determined in the Dex-PEG ATPSs quite significantly, whereas for proteins, the range of K-values in Dex-PEG ATPSs exceeded those in PEG-Na₂SO₄ ATPSs for three proteins, and were very similar for two proteins. This observation supported the notion that the ATPSs formed by two polymers are more suitable for protein analysis than those formed by a single polymer and a salt. The single polymer-salt ATPSs have an advantage for protein isolation/separation.

Keywords: Aqueous two-phase partitioning; Dex-PEG aqueous two-phase system; PEG-sodium sulfate aqueous two-phase system; Proteins; Salt and osmolyte additives; Small organic compounds.