BACKGROUND: This study focuses on obtaining a highly purified lysophosphatidylcholine from phosphatidylcholine powder via hydrolysis, catalyzed by a phospholipase A2 in a solvent-free medium, and subsequent purification either by organic solvent or by supercritical fluid extraction. RESULTS: A solvent-free hydrolysis reaction of phosphatidylcholine powder to produce lysophosphatidylcholine with residual phosphatidylcholine content lower than 10% has been described and scaled-up. Supercritical fluid mixtures of carbon dioxide, ethanol and formic acid were studied at 250, bar 50 degrees C to selectively extract the free fatty acids fraction from hydrolyzed phosphatidylcholine. Finally, a solid product with lysophosphatidylcholine content of c. 95% (w/w) and with free fatty acids content of c. 5% (w/w) was attained. Under optimum conditions, the content of the original phosphatidylcholine in the lysophosphatidylcholine product was lower than 5%(w/w). CONCLUSIONS: An efficient hydrolysis of phosphatidylcholine by phospholipase A2 in a solvent-free reaction was developed. In addition, three novel strategies for the purification of lysophosphatidylcholine have been developed: (1) hexane/formic acid extraction process, (2) supercritical CO2 extraction with ethanol as modifier, and (3) supercritical CO2 extraction with ethanol/formic acid as modifier. (C) 2012 Society of Chemical Industry