A triazacyclononane-ionic liquid (IL)-based sorbent was synthesized, characterized, and applied for rapid and inexpensive purification of hexahistidine tagged (His-tagged) proteins via liquid-liquid extraction. Selective partition of the target proteins between the IL phase and aqueous phase highly depends on a variety of parameters, such as the affinity ionic liquid (AIL), coordinated metal ions, and ionic strength. Compared with the chromatographic properties of Cu2+-AIL/IL and Ni2+-AIL/IL, Zn2+-AIL/IL exhibited relatively higher binding capacity to separate the proteins with high purity (up to 95%). A reclamation method for the AIL/IL extraction sorbent was further established by washing with ethylene diamine tetraacetic acid (EDTA) and then re-immobilization of metal ions. The purification strategies described herein were mild and suitable for a large scale separation of biologically active macromolecules with the possibility of continuous processing and a reduction in operational cost. (C) 2015 Elsevier B.V. All rights reserved.