Phase equilibrium and protein partitioning in aqueous two-phase systems containing imidazolium ionic liquids and surfactant at low voltage levels

In this paper, the binodal data of aqueous two-phase system (ATPS) containing imidazolium-based ionic liquids (chlorinated 1-ethyl-3-methylimidazole, [Emim]Cl), and a non-ionic surfactant (octylphenol polyethoxylene, Triton X-100) was obtained by means of the cloud point method in the temperature intervals T = (288.15-308.15) K. It was found that the [Emim]Cl/Triton X-100 ATPS was sensitive to temperature and had a long phase segregation time. Screening experiments of various additives showed that adding 7% of sodium dodecyl sulfate (SDS) to the total mass of surfactants (w/w) reduced the temperature sensitivity, and improved the phase separation rate of the system. The liquid-liquid equilibrium (LLE) data of [Emim]Cl/Triton X-100 ATPS without additive and containing 7% SDS at 298.15 K was further studied according to the cloud point method. To further shorten the long phase disengagement time, a low-voltage alternating current (AC) electric field was added to the [Emim]Cl/surfactant ATPS. Through the optimization of AC electric field conditions (such as electric field type, frequency and duty ratio), a relatively fast and stable [Emim]Cl/surfactant ATPS was established. In 0.67 V/cm field strength, 1000 Hz frequency and 50% duty ratio of impulse waveform, the phase splitting time is shortened by 2-3 times. With and without electric field, protein partitioning of cytochrome c (Cyt-c), bovine serum albumin (BSA) and lysozyme (Lys) in [Emim]Cl/surfactant ATPS indicated that Cyt-c and BSA tended to distribute in the bottom phase, while Lys was inclined to distribute in the top phase. The application of the AC electric field can still decrease the phase separation time in the protein distribution process, and has no significant effect on the distribution behavior of proteins. Furthermore, Lys activity in the top phase of ATPS was significantly enhanced, which could be caused by the low concentration of Triton X-100 in the top phase based on the comparison of Lys fluorescence spectra analysis in aqueous solution, in the top phase and in the bottom phase of [Emim]Cl/surfactant ATPS.