The effects of trans fatty acids on intestinal lipoprotein secretion were determined in polarized Caco-2 cells. Palmitic acid (16:0), palmitoleic acid (c-16:1 Delta 9), and palmitelaidic acid (t-16:1 Delta 9), as well as stearic acid (18:0), oleic acid (c-18:1 Delta 9), c-vaccenic acid (c-18:1 Delta 11), elaidic acid (t-18:1 Delta 9), and t-vaccenic acid (t-18:1 Delta 11) were studied. Compared with 18:0 (control), c- and t-18:1 Delta 9 increased triacylglycerol secretion (2.7- and 3.6-fold, respectively) as well as apolipoprotein (apo) B-48 and apo B-100 secretion (both 1.6-fold compared with 18:0); c- and t-18:1 Delta 11 caused a modest 1.7-fold increase in triacylglycerol secretion with no significant effect on secretion of apo B. Thus, the position of the double bond in the 18:1 isomers, but not its geometrical configuration, affected lipoprotein secretion by Caco-2 cells. In contrast, the effects of the geometrical isomers (cis and trans) of C-16 fatty acids were not comparable: t-16:1 Delta 9 did not affect triacylglycerol and apo B secretion (compared with 16.0, as control) whereas c-16:1 Delta 9 was a potent stimulator of secretion of triacylglycerol (2.4-fold higher than 16:0), apo B-48 (1.3-fold higher than 16:0), and apo B-100 (1.5-fold higher than 16.0). We conclude that the carbon chain length of fatty acids, as well as the position of double bonds and their stereochemical configuration, are important determinants of the unique effects of various species of dietary tr ails fatty acids on lipoprotein secretion and composition in Caco-2 cells.