Abstract: Extraction with aqueous two-phase system ( ATPS) is a novel separation technique, which shows some advantages including easy scale-up, mild conditions and continuous operation.The properties and applications of newly developed different ATPSs ( polymer-salt-water, organic solvent-salt-water, ionic liquid-salt-water, ionic liquid-surfactant-salt-water) in extraction of proteins, amino acids, antibiotics and other small molecules in herbs were reviewed. ATPS extraction combining with other technologies were summarized, and the prospect of ATPSs was also discussed.