Aims and Objectives: Separation and purification is a series of processes intended to isolate a single type of biomolecule from a complex mixture. Innovations in improvement of biodownstream processing, which is responsible for the separation of about 50–80% of recombinant proteins and other biomolecules, play a very important role in increasing the yield and reducing the cost of biopharmaceutical production. Methods: Biomolecule isolation and purification from a fermentation broth usually involve centrifugation, filtration, adsorption, and chromatography steps. Results and Discussions: Each step contributes to the product cost and product loss. Thus, we consider that solvent extraction and solvent sublation are the more economic processes for the separation of biomolecules. Conclusion: In extraction of phenylalanine, maximum extraction was observed at amino acid: surfactant ratio 1:1, amino acid: extractant ratio 1:1500, and pH at 3.1. The highest value of % recovery percentage and Co/Cw was 76.3 and 10.21, respectively. The main motive of this article is to provide the advantages of study on the solvent sublation and solvent extraction of l-phenylalanine over the other techniques.