Alterations in chylomicron and VLDL TAG and the magnitude of postprandial lipemia were studied in healthy volunteers after two meals of equal FA composition but different TAG FA positional distribution. Molecular level information of individual lipoprotein TAG regioisomers was obtained with a tandem MS method. The incremental area under the response curve of VLDL TAG was larger (P = 0.021) after modified lard than after lard. In plasma TAG, the difference did not quite reach statistical significance (P = 0.086). In general, there were less TAG with palmitic acid in the sn-2 position and more TAG with oleic acid in the sn-2 position in chylomicrons than in fat ingested. From 1.5 to 8 h postprandially, the proportion of individual chylomicron TAG was constant or influenced by TAG M.W. VLDL TAG regioisomerism was similar regardless of the positional distribution of fat ingested. Significant alterations were seen in VLDL TAG FA, in M.W. fractions, and in individual regioisomers with respect to time. The TAG sn-14:0-18:1-18:1 + sn-18:1-18:1-14:0, sn-16:0-16:1-18:1 + sn-18:1-16:1-16:0, and sn-16:1-18:1-18:1 + sn-18:1-18:1-16:1 decreased (P < 0.05); and sn-16:0-16:0-18:2 + sn-18:2-16:0-16:0, sn-16:0-16:0-18:1 + sn-18:1-16:0-16:0, sn-16:0-18:1-16:0, and sn-16:0-18:1-18:2 + sn-18:2-18:1-16:0 increased (P < 0.05) after both meals. In conclusion, positional distribution of TAG FA was found to affect postprandial lipid metabolism in healthy normolipidemic subjects.