Municipal sludge from wastewater treatment plants is a promising lipid feedstock for biodiesel production as it contains a significant amount of lipids. However, the energy necessary to remove its high water content is a major inconvenience for scaling up because of the high associated cost. In addition, the expensive conventional sludge drying methods are not effective enough for lipid recovery, thus reducing the potential biodiesel production. This study explores an alternative method, the direct sequential liquid-liquid extraction, which was performed in a batch mixer-settler reactor at room temperature, using hexane as a solvent, after previous sludge acidification showed significant increase in the lipid efficiency. The optimisation study demonstrated that, after three stages, 91% of lipid from primary sludge was recovered. The optimised extraction gave slightly higher lipid (27%, dry sludge) than the standard method (25%, dry sludge), supporting the suitability of the proposed process. Finally, this work demonstrates that the residual lipid-extracted sludge is still a good feedstock for energy production via anaerobic digestion. Anyway, the economic and environmental aspects of biodiesel production from sewage sludge could be improved. (C) 2014 Elsevier B.V. All rights reserved. ; Peer Reviewed ; Postprint (author’s final draft)