Non-monotonic dose effects of in utero exposure to di(2-ethylhexyl) phthalate (DEHP) on testicular and serum testosterone and anogenital distance in male mouse fetuses

Di(2-ethylhexyl) phthalate (DEHP) is a ubiquitous environmental contaminant. Epidemiological studies suggest that DEHP decreases masculinization of male fetuses. Numerous rat studies report DEHP reduces fetal testosterone production at doses greatly exceeding human exposure. We fed pregnant CD-1 mice 0.5–500,000 μg/kg/day DEHP from gestation day (GD) 9–18 and examined mothers and male fetuses on GD 18. We assessed non-monotonic dose–response by adding a quadratic term to a simple linear regression model. Except at the 500,000 μg/kg/day dose, DEHP stimulated an increase in maternal and fetal serum testosterone and increased anogenital distance (AGD). Non-monotonic dose–response curves were noted for AGD and maternal, and testis testosterone (P values 0.013–0.021). Because data from our highest dose (500,000 μg/kg/day) did not differ significantly from controls, this dose could have been incorrectly assumed to be the NOAEL had we only tested very high doses, as is typical in studies for regulatory agencies.

► DEHP is reported to alter male fetus masculinization and reduce anogenital distance. ► Most studies are of high-dose effects, beyond human exposure levels. ► We dosed pregnant mice with DEHP at 0.5–500,000 μg/kg/day. ► DEHP increased maternal and fetal serum testosterone and AGD at lower doses. ► No effects of DEHP were seen at the highest dose.