The absorption of folic acid in fortified white and whole-wheat bread, rice, or pasta or in solution was evaluated in human subjects with use of a single-dose, dual-label, stable-isotope protocol that did not involve prior loading of subjects with nonlabeled folate. In each of five sequential trials, 14 adults received a single oral dose of [13C5]folic acid in one of the four fortified cereal-grain products or in water concurrently with an intravenous injection of [2H2]folic acid. In two additional trials, subjects received oral [13C5]folic acid with or without a light breakfast meal. In all trials, urine was collected 24-36 h postdosing and the isotopic labeling of urinary folates determined. Isotope excretion ratios of urinary folates (% [13C5]folate dose/% [2H2]folate dose), which were used as criteria of absorption, indicated no significant differences among the various fortified foods and the control (P = 0.607). Because statistical power was sufficient to have detected a 50% difference from the control, these results suggest that [13C5]folic acid in these fortified cereal-grain foods was highly available. This study also suggests that fortification will contribute effectively to the folate status of the population. Consuming [13C5]folic acid after a light breakfast meal led to a small reduction in absorption relative to the control without food (P < 0.085). Between-subject variation in this protocol exceeded that observed in previous studies conducted using prior saturation of subjects with nonlabeled folic acid. We recommend that either prior saturation or multiple doses be used in future applications of this technique to improve precision.