EGG yolk phospholipids have a great interest in medical and food medical and food industry due to their biophysical and biological biophysical and biological characteristics. The present study extracts, crystallizes and evaluates the egg yolk phospholipids belonging two different species of birds, hen and duck. Phospholipids were extracted by solvent extraction technique, purified with silica gel, and crystallized by slow evaporation method. Egg yolk phospholipids were characterized by using high pressure liquid chromatography (HPLC), XRD, FT-IR, SEM and TEM spectroscopy. Extracted phospholipids exhibited high purity and identical chemical groups but XRD, HPLC, and TEM results showed significant differences between hen and duck egg yolk phospholipids. Therefore, these results confirm that the species of birds has a more significant effect on the composition and biophysical characteristics of extracted egg yolk phospholipids, emphasizing the importance to control individual parameters for successful using of phospholipids for their specific applications as liposomes for drug delivery.