An environment-friendly reactive extraction method as a novel phase separation technique is considered to extract Lactic Acid (LA) from the aqueous solution by means of environmentally benign green diluents along with a synergistic mixture of extractants. Reactive extraction of LA was carried out through a synergistic mixture of extractants (trioctylamine (TOA), Aliquat336, and tridodecylamine (TDDA)), organic solvents, and non-toxic and biocompatible green diluents. According to the various investigated LA reactive extraction processes, the process featuring sun ower oil, synergist extractant (TOA and Aliquat336), and Butan-2-ol was found to be the most favorable system. The LA distribution coe cient (K D = 27:75 0:22) and extraction e ciency ( = 97:17 0:54%) were obtained by a system consisting of LA concentration (0.05 [M]), TOA (15%, v/v), Aliquat336 (15%, v/v), phase ratio (1:1, v/v), solvent ratio (2.5, v/v), agitation speed (100 rpm), and stirring time (120 min). The main driving force for this research work on the LA reactive extraction from the aqueous solution is the development of a suitable combination of extractants, organic solvents, and natural solvents that possesses high a nity with LA and, also, has lower toxicity with respect to LA-producing microbes.