Normal, healthy, free-living adults ingested either 18 g/d olestra, with or without 1.1 mg tocopheryl acetate/g olestra, or 18 g/d triglyceride placebo, for 16 wk in a double-blind, placebo-controlled study. Serum concentrations of alpha-tocopherol, alpha-carotene, beta-carotene, lycopene, lutein/zeaxanthin, retinol and cholesterol were measured biweekly. Serum 25-hydroxyvitamin D concentration, prothrombin time, partial thromboplastin time and plasma concentration of functional prothrombin (Simplastin-Ecarin assay) were measured at wk 0, 8 and 16. Relative to the placebo group, serum alpha-tocopherol concentration was reduced 6% for the group given 18 g/d olestra. Addition of tocopheryl acetate to olestra partially offset the effect of olestra. For the group given 18 g/d olestra plus 1.1 mg tocopheryl acetate/g olestra, serum alpha-tocopherol concentration was 4% less than the placebo value. Olestra reduced serum concentration of beta-carotene by 27%; the other carotenoids were similarly affected. Serum cholesterol concentration was reduced approximately 4.5% in the olestra groups, relative to placebo, but the differences were not significant. Serum triglycerides, serum 25-hydroxyvitamin D, prothrombin time, partial thromboplastin time or the plasma concentration of under-gamma-carboxylated prothrombin were unaffected by olestra. Clinical observations and laboratory measures indicated no health-related effects of olestra; mild-to-moderate transient gastrointestinal symptoms such as bloating, cramping, loose stools and diarrhea were reported by all groups.