Pharmacokinetic studies of the disposition of acetaminophen were carried out in 12 chronically catheterized pregnant sheep. Acetaminophen was infused separately into the mother and the fetus on two occasions to steady state. The transplacental clearances from both sides of the placental membrane and the nontransplacental clearances in both the mother and the fetus were then determined using an indirect method, a two-compartment open pharmacokinetic body model. The placental clearance and the nontransplacental clearances were also determined independently by direct methods by measuring the extraction ratio and urinary clearances in both the mother and fetus. Model dependent (indirect) and model independent (direct) methods yielded similar results. Neither acetaminophen glucuronide nor acetaminophen sulfate was transferred across the placenta. However, acetaminophen was transferred in both directions exclusively via a passive diffusion mechanism with an extraction ratio of 0.12 across the placenta. Neither glucuronidation nor sulfation capacity was found in the sheep placenta. However, the conjugation pathways accounted for nearly all the nontransplacental elimination of acetaminophen in the mother. Both transplacental clearance and nontransplacental clearance of acetaminophen in the fetus increased with gestational age. The glucuronidation and sulfation activities were determined in the fetus in utero and found to be 16 and 74% of the respective activities in the pregnant ewe on a body weight basis. The two conjugation clearances did not increase significantly with the gestational age of the fetus during the third trimester.