These days, the environmental issues from the usage of fossil fuels have been widely highlighted since it is high in demand to come up with the mitigation actions. These environmental issues may be derived from the extensive pollution and release of greenhouse gas (GHG) emissions especially from the industry and transportation sector. GHG will trap the heat in the atmosphere which leads to global warming. Therefore, the usage of microalgae as biomass for the feedstock can help in tackling these issues. As a feedstock that is common in marine environment, microalgae can be converted into high-quality biomass with its accelerated metabolism. In addition, microalgae can rapidly achieve optimal growth and harvesting conditions for further usage on generating renewable energy in the form of liquid or gaseous fuels. Besides, the extraction of lipid from microalgae biomass was widely studied. Specifically, lipid extraction is currently being innovated and modified in order to meet the sustainability aspects. Prior to lipid extraction, pre-treatment method can be considered as an essential step to boost the amount of lipid produced. Thus, the effect of pre-treatment method called ultrasonication before lipid extraction from dry biomass (Tisochyris Lutea, T-iso) has been discussed. Dry T-iso did undergoes ultrasonication water treatment and followed by lipid extraction. Methanol: chloroform (2:1) was used as the solvent system for control during extraction. The amount of lipid produced while adding the pre-treatment method before extraction has also been determined and compared with the conventional method. 2-Methyltetrahydrofuran (2-MeTHF) was used as the solvent system in this due to its green properties. Research. An addition parameter of isoamyl alcohol was also used to compare the effect of this solvent with 2-MeTHFFrom this research, the lipid yield without any pre-treatment method for chloroform:methanol (2:1 v/v), 2-MeTHF and 2-MeTHF:isoamyl alcohol (2:1 v/v) were 20.33%, 20.67% and 30.00%. Meanwhile, the lipid yield with ultrasonication pre-treatment method for chloroform:methanol (2:1 v/v), 2-MeTHF and 2-MeTHF:isoamyl alcohol (2:1 v/v) were 22.67%, 31.00% and 36.67%. In a nutshell, the addition of pre-treatment method in this study will help to boost the lipid efficiencies and as well as tackling the environmental concern by achieving a more sustainable process on producing biodiesel.