The concentration of betamethasone has been measured in maternal peripheral plasma, umbilical cord artery and vein, and amniotic fluid following maternal administration of betamethasone phosphate and betamethasone acetate on 3 consecutive days. Betamethasone was measured by radioimmunoassay following column chromatography. The findings show that betamethasone is transferred across the human placenta, circulates in the fetus and appears in amniotic fluid. During the 3 days after the start of treatment, levels of the betamethasone were similar in maternal and umbilical cord blood and in amniotic fluid. Thereafter, although levels in the mother were measurable for up to 7 days after the initial injection, the drug was detected in the cord plasma of only one baby. In vitro incubation studies of human placental tissue with 3H betamethasone identified 11-keto betamethasone as the major metabolite of betamethasone.