Many industries are using directly biomass as a raw material for the production of valuable biochemicals. These are used different toxic chemicals such as sulphuric acid, hydrochloric acid, nitric acid, etc. as a catalyst during the production of biochemicals and exit downstream waste material and create environmental pollution. Therefore, downstream waste material separation is important for decrease environmental pollution. Levulinic acid is a carboxylic acid which can be present downstream of industrial waste. It is a very important chemical and can be transformed into different important chemicals such as 1–4 pentanediol, aminolevulinic acid, succinic acid, gamma valarolactone, hydoxyvaleric acid, diphenolic acid, etc. It is considered in the top ten important and bio-derived acids. Levulinic acid can be directly produced from biomass through chemical synthesis and fermentation processes at industrial and lab scales. The synthesis process produces the char whereas the fermentation process produces wastes during the production of levulinic acid, thus increasing the production cost and industrial waste downstream. The separation of levulinic acid from the waste is costly and challenging. In this study, reactive extraction was employed with TOA in i-octanol for the separation of levulinic acid. The experimental results have been expressed in various performance parameters like distribution coefficient (0.099–6.14), extraction efficiency (9–86%), loading ratio (0.09–0.7), and equilibrium constant (11.34–1.05). The mass action law model was also been applied and found the predicted value close with the experimental results. The mixer settler extraction in the series was used to achieve more than 98% separations of acid. Further, the conceptual approach for levulinic acid separation using mixer-settler reactor scheme is discussed and presented various design parameters including extraction efficiency, diffusion coefficient, extraction equilibrium constant, and loading ratio. This study decreases the industry downstream material and can be solved environmental pollution problems.