An industrially relevant method for disrupting microalgal cells and preferentially extracting neutral lipids for large-scale biodiesel production was demonstrated on pastes (20-25% solids) of Nannochloropsis sp. The highly resistant Nannochloropsis sp. cells. were disrupted by incubation for 15 h at 37 degrees C followed by high pressure homogenization at 1200 +/- 100 bar. Lipid extraction was performed by twice contacting concentrated algal paste with minimal hexane (solvent:biomass ratios (w/w) of <2:1 and <1.3:1) in a stirred vessel at 35 degrees C. Cell disruption prior to extraction increased lipid recovery 100-fold, with yields of 30-50% w/w obtained in the first hexane contact, and a further 6.5-20% in the second contact. The hexane preferentially extracted neutral lipids over glyco- and phospholipids, with up to 86% w/w of the neutral lipids recovered. The process was effective on wet concentrated paste, required minimal solvent and moderate temperature, and did not require difficult to recover polar solvents. (C) 2013 Elsevier Ltd. All rights reserved.