Trans FA (TFA) have at least one trans double bond and comprise several isomers and types, including many of the CLA (e.g., c9,t11-18:2 CLA). Some TFA may have adverse effects (e.g., cardiovascular disease), whereas some are thought to have beneficial effects (e.g., anticarcinogenicity). The presence of TFA in human tissues and fluids is related to dietary intake, although this relationship is not completely understood--especially in regard to serum lipid fractions. This study was conducted as part of an investigation designed to test the influence of butter (B), "low TFA" margarine (LT), and regular margarine (RM) on milk fat content. Here we tested the secondary hypothesis that consumption of B, LT, and RM by lactating women would result in differential distribution of TFA and CLA in major serum lipid classes. Breast-feeding women (n = 11) participated in this randomized Latin-square study consisting of five periods: intervention I (5 d), washout I (7 d), intervention II (5 d), washout II (7 d), and intervention III (5 d). Extracted serum lipid was separated into cholesterol ester (CE), TAG, and phospholipid (PL) fractions and analyzed for total and isomeric TFA and CLA concentrations. Data indicate that TAG consistently contained the highest concentration of total t-18:1. No interaction between treatment and fraction was found for any of the t-18:1 isomers identified. Absolute concentration of each t-18:1 isomer was greatest during the RM period, regardless of fraction. On a relative basis, concentrations of t10-18:1 and t12-18:1 were most responsive to treatment in the CE fraction. The concentration of c9,t11-18:2 CLA was highest in the TAG fraction and lowest in the PL fraction, regardless of treatment. In summary, these results indicate (i) that there is a differential distribution of some isomeric TFA and CLA among human serum lipid fractions and (ii) that dietary TFA intake influences absolute and relative concentrations of some of the isomers in selected fractions.