Various solvents, including water, hexane, methanol, and chloroform/methanol (1:1 v/v), were tested to identify the efficiency of lipid extraction from Trichosporon oleaginosus and an oleaginous fungal strain SKF-5 under ultrasonication (520kHz 40W and 50Hz 2800W) and compared with the conventional chloroform methanol (2:1 v/v) extraction method. The highest lipid recovery 10.2% and 9.3% with water, 43.2% and 33.2% with hexane, 75.7% and 65.1% with methanol, 100% and 100% w/w biomass with chloroform/methanol were obtained from T. oleaginosus and SKF-5 strain, respectively, at ultrasonication frequency 50Hz and power input 2800W. Ultrasonication chloroform/methanol extraction recovered total lipid in a short time (15min) and low temperature (25°C). Whereas the conventional chloroform methanol extraction to achieve total lipid recovery required 12h at 60°C. Ultrasonication chloroform/methanol extraction would be a promising method of lipid extraction from the microorganisms.