The aim of the study was to determine the impact of the frying process on the fractional composition of AME Biodiesel in comparison to the AME obtained from unused (fresh) animal fat. The fresh animal fat was divided into two portions. One was used for frying chips at 63°C for a period of 4 hours. The study showed the AME biodiesel produced from unused (pure) animal fat generally has better distillation properties. The temperatures at the start of distillation were similar for both of the AMEs. Within the 45-60% mid-range temperatures, the AME produced from the used cooking animal fat was characterized by higher distillation temperatures for the same volume of fuel. The largest differences were observed for the 85% and 95% distillation temperatures and the final temperature of the distillation process. This may testify to lower purity of the AME produced from the used cooking oil. In such a biofuel may consist less volatile mono-diglycerides or other chemicals which remain in the oil after frying. It must be said, though, these are not solid particles, as those were separated from the oil through filtration.