According to the United Nations Food and Agriculture Organization (FAO), Brazil figures as one of the world's largest producers of fish with an estimated 20 million tons in 2030. In the state of Ceará the production of Nile Tilapia (Oreochromis niloticus) is particularly important. Through the consumption and preparation of fish, there are parts of it that are not used and are often discarded. A major part of the fish, the gut, has oil that is able to become a raw material for biodiesel production. In this sense, the present work aimed to obtain a methodology for the treatment of a fish oil stored for approximately 18 months and the subsequent production of biodiesel, focusing on reducing the acidity index, which is a problem for fuels generated from this product. The standard set forth in ANP Resolution No. 45/2014 of the National Agency of Petroleum, Natural Gas and Biofuels was followed. After treatment, fish oil showed reduction of acidity about 97%.