The use of lipids obtained from microalgae biomass has been described as a promising alternative for production of biodiesel to replace petro-diesel. It involves steps such as the cultivation of microalgae, biomass harvesting, extraction and transesterification of lipids. The purpose of the present study was to compare different methods of extracting total lipids. These methods were tested in biomass of Chlorella vulgaris with the solvents ethanol, hexane and a mixture of chloroform:methanol in ratios 1:2 and 2:1. The solvents were associated with other mechanisms of cell disruption such as use of a Potter homogenizer and ultrasound treatment. The percentage of triglycerides in the total lipids was determinated by the glycerol-3-phosphate oxidase-p-chlorophenol method (triglycerides monoreagent K117; Bioclin). Among the tested methods, the mixture of chloroform:methanol (2:1) assisted by ultrasound was most efficient, extracting an average of 19% of total lipids, of which 55% were triglycerides. The gas chromatographic analysis did not show differences in methyl ester profiles of oils extracted under the different methods.