Accelerated solvent extraction (ASE), a commercially available pressurized fluid extraction technique and conventional manual extraction were compared to identify the most effective chloroform?methanol extraction method for algal lipids. Using optimal ASE operating conditions (methanol/chloroform = 2 : 1 by vol, 100 �C, static time of 5 min, and four static cycles), the lipid contents of Chlorella vulgaris, C. sorokiniana, C. zofingiensis and Nannochloropsis gaditana were 27.5%, 25.8%, 15.2%, 29.8% of dry biomass, respectively. Here, the total fatty acid methyl ester (FAME) content of dry biomass from ASE extraction was found to be 1.3?2.7 fold higher than that from conventional manual extract from these species, demonstrating that ASE exhibited significant improvement for lipid and FAME recovery. Furthermore, ASE showed the capacity to extract all-cis-5,8,11,14,17-eicosapentaenoic acid (EPA) as 3.0% of dry biomass from Nannochloropsis gaditana suggesting that ASE has the potential to obtain polyunsaturated fatty acids (PUFAs) as well.