Biofuel from microalgae has been gaining attention due to the depletion of fossil fuel and the related environmental issues. Numerous researchers have been focusing on the efficient extraction of the lipid from the microalgae or direct conversion into fuel forms. However, there have been few studies on refining the microalgae oil, even though refining process is necessary to use the oil as feedstock for the subsequent conversion. The refining process is expected to eliminate the impurities, especially phosphorus. In this study, lipid extraction using organic solvent and refining process were investigated to efficiently produce phosphorus-free lipids from microalgae. Aurantiochytirum sp. KRS101 was selected, and GC-FID, TLC, and ICP-OES were used to qualify the extracted lipids.