This work gives an outline of current research patterns in bio-oil separation techniques and their applications. Due to unpredictable nature of bio-oil, various challenges have been reported in downstream operations. Several methods have been presented to redesign the bio-oil by separating the combination of bio-oil compounds into a group of chemical substances and portions advanced in definite groups of chemical mixtures. In this review, different separation strategies for bio-oil such as fractionation, water extraction, organic solvent extraction, supercritical fluid extraction, and membrane separation, with the qualities of bio-oil extracts and their applications have been discussed. It was found that the most favorable techniques are fractionation and organic solvent extraction. The present review focuses on recent developments made from model mixtures present in bio-oil as well as pyrolysis bio-oil; techno-economical analysis has been presented for researchers who are expected to bridge knowledge gap and for industrial practitioners.