The present work aims to separate an amino acid (l-phenylalanine) from a highly dilute aqueous solution by the 'adsorptive bubble separation method'. Separation is based on the principle of adsorption at the interphase of the gas bubble and aqueous phase and subsequent amino acid extraction in the top layered nonaqueous phase. The operation was carried out in a long glass column with a comprehensive solvent chamber at the top. Design-Expert software optimizes the process parameters, and the process involves three variables: gas (nitrogen) flow rate, extractant, and solvent quantity, which control responses, recovery percentage, and the ratio of enrichment. The design models are found to be significant. The maximum recovery of amino acid observed was 75.545% and the value of the enrichment ratio was 3.87. In addition, two models were generated by the software, which was statistically significant (P < 0.05). Therefore, this process is adopted as it is of straightforward construction, eco-friendly, and less costly than other methods previously known to recover l-phenylalanine from a dilute solution.