The aim of this work was to study the ultrasonication-assisted Liquid Tri-phasic Flotation (LTF) System to obtain lipid and protein from microalgae Chlorella sorokiniana in a single step as a novel process. In the current study, biorefinery of Chlorella sorokiniana was performed using LTF system in a single step. The highest protein recovery of 97.43 ± 1.67 and lipid recovery of 69.50 ± 0.54 were obtained. The corresponding parameters were microalgae biomass loading of 0.5 w/v, ammonium sulphate concentration of 40 w/v, volume ratio of 1:1.5 (salt:alcohol), ultrasonication pulse mode of 20 s ON/20 s OFF at 20 amplitude for 5 mins, flotation air flowrate of 100 mL/min. Additionally, recycling of alcohol phase to study the circular nature of proposed biorefinery was investigated. The proposed LTF system for extraction of proteins and lipid reduces the number of operation units required in this biorefinery approach. © 2020 Elsevier Ltd