Biomolecules, specifically proteins, polysaccharides, and secondary metabolites are potential lead compounds due to their remarkable pharmacological properties. However, the complex molecular structure of the biomolecules makes their separation processes of great challenges. The conventional downstream processes require multistep protocols that are less efficient, high solvent consumption, expensive, time-consuming, and laborious. Hence, aqueous two-phase system (ATPS) is a reliable technique for the extraction and purification of biomolecules from a complex mixture. ATPS is an environmentally friendly, simple, cost effective, and easily scalable process. It requires a short processing time to separate biomolecules of industrial values simultaneously in a single process. Modifications have also been performed by introducing deep eutectic solvents, ionic liquids, carbohydrates, amino acids or copolymers to enhance the process efficiency with an increased yield, purity and bioactivity of recovered biomolecules. This review attempts to review the recent developed ATPSs and their efficiency to extract, isolate, and purify biomolecules such as proteins, polysaccharides, secondary metabolites and other biological substances. The review provides insights into the feasibility and reliability of ATPS for biomolecule recovery.
The objective of the study was to fractionate the crude extract of Eurycoma longifolia (E. longifolia) roots and identify the intense peaks using HPLC-PDA-MS/MS, UPLC-MS/MS and H-NMR. Column chromatography was used to fractionate the crude extract into individual fractions using six solvent systems ranged from ethyl acetate, methanol and water in increasing polarity. Two fractions with nearly pure and intense peaks were selected for compound identification. Chromenone (coumarin) and chromone derivatives were putatively identified, besides several previously reported quassinoid glycosides (eurycomanone derived glycoside, 2,3-dehydro-4α-hydroxylongilactone glucoside, eurycomanol glycoside and eurycomanol trimer) in the fraction 11 of 100% methanol. A newly reported compound, namely hydroxyl glyyunanprosapogenin D (838 g/mol) was proposed to be the compound detected in the fraction 11 of 50% ethyl acetate and 50% methanol. This is also the first study to report the identification of chromenones and chromones in E. longifolia extract.