Aqueous two-phase system

Aqueous biphasic systems (ABS) or aqueous two-phase systems (ATPS) are clean alternatives for traditional organic-water solvent extraction systems. Aqueous biphasic systems (ABS) or aqueous two-phase systems (ATPS) are clean alternatives for traditional organic-water solvent extraction systems. ABS are formed when either two polymers, one polymer and one kosmotropic salt, or two salts (one chaotropic salt and the other a kosmotropic salt) are mixed at appropriate concentrations or at a particular temperature. The two phases are mostly composed of water and non volatile components, thus eliminating volatile organic compounds. They have been used for many years in biotechnological applications as non-denaturing and benign separation media. Recently, it has been found that ATPS can be used for separations of metal ions like mercury and cobalt, carbon nanotubes, environmental remediation, metallurgical applications and as a reaction media. In 1896, Beijerinck first noted an 'incompatibility' in solutions of agar, a water-soluble polymer, with soluble starch or gelatine. Upon mixing, they separated into two immiscible phases. Subsequent investigation led to the determination of many other aqueous biphasic systems, of which the polyethylene glycol (PEG) - dextran system is the most extensively studied. Other systems that form aqueous biphases are: PEG - sodium carbonate or PEG and phosphates, citrates or sulfates. Aqueous biphasic systems are used during downstream processing mainly in biotechnological and chemical industries. It is a common observation that when oil and water are poured into the same container, they separate into two phases or layers, because they are immiscible. In general, aqueous (or water-based) solutions, being polar, are immiscible with non-polar organic solvents (chloroform, toluene, hexane etc.) and form a two-phase system. However, in an ABS, both immiscible components are water-based. The formation of the distinct phases is affected by the pH, temperature and ionic strength of the two components, and separation occurs when the amount of a polymer present exceeds a certain limiting concentration (which is determined by the above factors). The 'upper phase' is formed by the more hydrophobic polyethylene glycol (PEG), which is of lower density than the 'lower phase,' consisting of the more hydrophilic and denser dextran solution. Although PEG is inherently denser than water, it occupies the upper layer. This is believed to be due to its solvent 'ordering' properties, which excludes excess water, creating a low density water environment. The degree of polymerization of PEG also affects the phase separation and the partitioning of molecules during extraction. ABS is an excellent method to employ for the extraction of proteins/enzymes and other labile biomolecules from crude cell extracts or other mixtures. Most often, this technique is employed in enzyme technology during industrial or laboratory production of enzymes.