The phase separation behaviors of PEG1000/sodium citrate, PEG4000/sodium citrate, PEG1000/ammonium sulfate, and PEG4000/ammonium sulfate aqueous two-phase systems were investigated, respectively. There are two distinct situations for the phase separation rate in the investigated aqueous two-phase systems: one state is top-continuous phase with slow phase separation rate and strong bottom-continuous phase with fast phase separation rate and weak volume ratio dependence. The system properties such as density, viscosity, and interfacial tension between top and bottom phases which have effects on the phase separation rate of aqueous two-phase systems were measured. The property parameter differences between the two phases increased with increasing tie line length and then improved the phase separation rate. Moreover, a modified correlation equation including the phase separation rate, tie line length, and physical properties of the four aqueous two-phase systems has been proposed and successfully tested in the bottom-continuous phase, whose coefficients were estimated through regression analysis. The predicted results of PEG1000/sodium citrate aqueous two-phase systems were verified through the stationary phase retention in the cross-axis countercurrent chromatography.
This paper presents the development of a novel cross-axis countercurrent chromatographic instrument (CCCI) with six separation columns, including design, dynamics, optimization, prototype construction, and experimental validation. The conceptual design and the structural design of the cross-axis CCCI are performed while considering the requirements for the separation operation and design. The dynamic analysis is carried out in order to guarantee the local balance and the global balance for the CCCI. The relationship between the mechanical structure parameters and partition efficiency is investigated by analyzing the effect of mechanical structure parameters on the centrifugal force field. By virtue of the modal analysis of the mechanical structure, the critical speed and the weak link of the CCCI are achieved. Aiming at the problem of the weak link, the structural optimization is done. The presented CCCI has six separation columns distributed around the central revolution axis, and it has more separation columns than that of the existing chromatographic instrument. The CCCI has an axial symmetry structure in the three-dimensional space. Thus it can have better dynamics stability than the CCCI having less separation columns with a symmetry structure in the plane. A physical prototype is built, and then the partition efficiency and its effect factors are tested by the single experimental variable method. The results show that the CCCI runs smoothly and has a good retention rate of stationary phase. It is also proved that the developed CCCI has a good partition efficiency on bovine serum albumin and lysozyme.
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