A novel class of aqueous biphasic systems (ABS) formed by zwitterions (ZI) has been investigated in the present work. A series of water soluble ZIs have been synthesized using triethylamine, N-Methylimidazole, N-Vinylimidazole, pyridine, N-Methylpyrrolidine, N-Ethylpiperidine and 1,4 butane sultone. The synthesized ZIs were explored for their ability to form biphasic systems in combination with aqueous inorganic salt solutions of K3PO4, K2HPO4 and K2CO3. The phase diagrams for all systems have been constructed through cloud point titration method at 298.15 K and atmospheric pressure. The phase behavior of ZI based ABS have been analyzed to understand the structural effects of ZIs as well as the effect of nature of salt used on the overall phase formation. Further the temperature dependence of the ZI based ABS was also explored by studying the phase behavior at variable temperatures of 298.15, 308.15 and 318.15 K. In order to estimate the applicability of proposed ZI based ABS, extraction experiments have been performed for an alkaloid i.e. caffeine for all synthesized ZIs with K3PO4 and at 298.15 K. ZI based ABS have been found to be capable of single step extraction of caffeine similar to IL based ABS thus providing the possibilities to explore these ZI based ABS as efficient extraction and separation systems. © 2019 IUPAC & De Gruyter. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. For more information, please visit: http://creativecommons.org/licenses/by-nc-nd/4.0/ 2019.