Rendang is a traditional Minangkabau cuisine with the main ingredient of beef or buffalo meat which is cooked using coconut milk and some spices. Rendang is cooked more than 2 hours, and after that do the repeated heating. This study aimed to determine the effect of repeated heating on fatty acid profiles of beef and spices of rendang. The analysis carried out at Chemistry Lab. and Biochemistry Lab. In Fateta Unand and Integrated Lab in IPB Bogor. This study designed used a completely randomized design with 5 treatments and 3 replications. The treatments that used in this study were A (raw beef), B (freshly cooked rendang), C (first heating), D (second heating), E (third heating) where the heating was done every 2 days. Data were collected for fatty acids profile of beef and spices of rendang. Analysis of the Type of Fatty Acid by HPLC method AOAC. The results shown that repeated heating of beef and spice of rendang had significantly different effect on the type of fatty acids and an increased trans fatty Acid of rendang.