The purification of phosphatidylcholine (PC) from the de-oiled lecithin was carried out using different physical separation techniques, such as extraction, precipitation, adsorption and column chromatography. The extraction of the PC from the de-oiled lecithin was carried out using different solvents. The effect of lecithin to solvent ratio on extraction efficiency was studied. It was found that the lecithin to solvent ratio of 1:7 gives higher percentage purity of PC. The precipitation experiments were carried out using acetone and magnesium sulphate hepta-hydrate (MgSO4 center dot 7H(2)O) of different concentrations and the effect of different concentrations of MgSO4 center dot 7H(2)O on percentage purity of PC was studied. Further enrichment of PC was done using column chromatography to increase its percentage purity. Different types of resins were used and the most suitable resin was selected for the column chromatographic purification. The percentage purity of the PC was increased to 84% after column chromatography. The new isocratic method for the HPLC analysis was developed for phosphatidylcholine. (C) 2010 Elsevier B.V. All rights reserved.