Enantioseparation of Phenylsuccinic Acid Enantiomers Using Aqueous Two-Phase Flotation and Their Determination by HPLC and UV Detection

Aqueous two-phase flotation (ATPF) is especially applicable to bioseparation. However, there is no previous work reporting on its application in enantioseparation. Using phenylsuccinic acid (H₂A) as the model enantiomers, ATPF was first introduced into the field of chiral separation. The contributions of a series of ATPF systems to the enantioseparation were investigated. The results indicated that an appropriate increase in the amount of phase-forming components and a decrease in pH values (from 5.5 to 2) are both beneficial for the enantioseparation. Enantioselective flotation and partitioning behavior are mainly dependent on pH values of the solutions, types, and concentrations of chiral selectors. Furthermore, salt, PEG, flow rates of air, and flotation time also have some effects on the enantioseparation. Under the optimal conditions, the enantioselectivity was calculated in terms of the separation factor (α) and enantiomer excess (e.e. %) as 1.99 and 23.49 %, respectively. Finally, the most reasonable mechanisms for H₂A enantioseparation in ATPF system have been proposed. An ATPF system composed of PEG2000/(NH₄)₂SO₄was proved to have the best ability for the separation of H₂A enantiomers. The explorations in our study will further enrich the enantioseparation methods and pave the way for the application of ATPF in the enantioseparation field.