Combinations of three cell disrupting methods were used for lipid extraction from the Gram-positive bacterium, Streptomyces pristinaespiralis: lysozyme treatment, freeze-drying and bead mill. The determination of extracted phospholipids (PL) was carried out using thin layer chromatography coupled with a flame ionisation detector. It appeared that freeze-drying induced the highest PL extraction and that only lysozyme addition permitted to extract phosphatidylserine. The optimal cellular dry weight for the extraction and the subsequent separation and analysis of the PL was 50 mg. (c) 2006 Elsevier Ltd. All rights reserved.