In this study, we examined the effects of dietary 1,3-diacylglycerol (DG) compared to conventional triacylglycerol (TG) oil on the postprandial response of total and chylomicron TG, glucose, insulin, and free fatty acid (FFA). This study was conducted using a cross-over design. Ninety subjects participated in the high-fat meal tolerance test where they were randomly assigned to consume two experimental sandwiches containing mayonnaise with TG or DG oil with a seven-day interval. Blood samples were collected before ingestion and at 2, 3, 4 and 6 hr time point after ingestion and analyzed for total and chylomicron TG, glucose, insulin, FFA and phospholipid fatty acid composition. Both TG and DG ingestion had similar effects on postprandial TG response, but a different response from chylomicron TG. Compared with the TG group, TG levels were significantly lower only at 6 hr time point in the DG group. On the other hand, chylomicron TG rose steeply at 2 hr time point and decreased faster in this group. Also, the adjusted value to fasting levels was the same as the unadjusted level. Fasting levels and net differences in insulin were significantly lower at 3 hr time point where chylomicron TG levels were significantly lower in the DG group. But those of glucose and FFA in the TG and DG groups did not differ significantly. Fasting and postprandial levels of fatty acid composition in serum phospholipids in the two groups did not differ significantly. In conclusion, this study indicated that one could reduce the magnitude of postprandial lipemia without influencing glucose metabolism by consumning DG oil as a substitute for TG oil. Based on the correlation of coronary artery disease and postprandial lipemia, dietary DG ingestion might have a beneficial effect in treating such a disease. Further studies are required to clarify the long-tenn effects of dietary DG on blood lipid levels in humans.