This study evaluated the effects of Dioscorea batatas and Gastrodia rhizoma on fatty acid composition of serum, liver and brain in rats which were fed control diet adding 5%, 10%, 15% Dioscorea batatas powder and 5%, 10%, 15% Gastrodia rhizoma powder for 3 weeks, respectively. In the other total fatty acid composition of serum lipids, the percentage of SFA was higher in the control group than in the other groups. The percentages 18:2 and 20:4 in serum lipids were significantly higher in the group 5, 6 and 7 than control group. In phospholipid fatty acid composition of serum lipids, the percentage of 18:2 was significantly higher in the group 5, 6 and 7 than in the other groups. 16:0, 18:0, 18:1, 18:2 and 20:4 were the main fatty acid in the total fatty acid and phospholipid fatty acid composition of liver lipids. 16:0, 18:0, 18:1, 18:2, 20:4 and 22:6 were the main fatty acid in the total fatty acid and phospholipid fatty acid composition of brain lipids.