Background: Liposomes have been widely studied in the field of medicine in recent years for they can reduce side effects and regulate drug release by delivering active ingredients to target tissues and cells through active or passive routes. There are many types of methods for formulating liposomes, and comparing those methods and choosing the most suitable one will allow the delivery of active ingredients and their incorporation into pharmaceutical forms, so this research was carried out. Aim: To compare liposome formation methods. Materials and Methods: The research work was carried out with the support of “Drug Design Laboratory” and “Pharmaceutical Analysis Laboratory” of the School of Pharmacy of MNUMS. To formulate liposomes, 5 different concentrations of samples were prepared using thin-film hydration, ethanol injection, and heating methods. Phospholipid concentration and light absorption were determined by spectrophotometer in each sample. The size of the resulting liposome was determined using a Nanophox. Results: Phosphatidilcholines were dissolved in distilled water at concentrations of 5, 10, 20, 30, 40, and 50 μg/ml and a standard curve was established by spectrophotometry to determine the concentration of phospholipids in liposome samples. The highest formulation was 76.98% when 75 mg of phospholipid was used in the preparation by thin film hydration method, when 50 mg of phospholipid was used in the preparation by ethanol injection method, the highest formulation was 85.17%, and for when 50 mg of phospholipid was used in the preparation by heating method, the highest formulation was 58.45%. The mean liposome size by ethanol injection method was 115 nm. Conclusions 1. Ethanol injection method for liposome formulation is more efficient compared to thin-film hydration method and heating method. 2. Liposome size was 115 nm when prepared by ethanol injection.