The bioavailability of diacylglycerol (DAG) microemulsion (DAGM) was investigated in rats. The fatty acid (FA) concentrations in the serum of all rats at 0, 6 and 24h and in feces after oral administration of DAGM and DAG oil were examined. The results showed that the FA concentration of the DAGM group was significantly higher than that of the DAG oil group at 6 h after treatment. The digestibility was detected in rats for 2 weeks. During this period, there was no significant difference in the body weight between the two groups. However, the total food consumption of rats treated with DAGM was evidently less than that treated with DAG oil (P = 0.006). The fat absorption of DAGM was significantly higher than that of DAG oil. The contents of 18:1, 18:2n–6 and 18:3n–3 were significantly lower in the DAGM group than that in the DAG group in fecal lipids. In conclusion, DAGM seems to be a good candidate for enhancing the bioavailability of DAG.