Xenobiotic estrogens in the environment or diet have received much attention as a possible source of certain hormonal disease states in human and wildlife. Therefore, the detection of estrogenic activity of any substance, especially those related to the food industry, is important. The estrogenic activity of p-hydroxybenzoic acid (PHBA), a compound related to a commonly used group of preservatives in food, cosmetic, and pharmaceutical preparations, was evaluated with immature and adult ovariectomized female mice (CD1) using two well-known bioassays. Subcutaneous administrations (sc) of different doses of PHBA were compared with estradiol (E-2), and their effects on vaginal cornification and uterotrophic activities were evaluated. Different groups of animals were treated sc daily for 3 days with vehicle (corn oil, 0.3 ml/100 g), E-2 (1 mu g/100 g), and PHBA (0.5, 5, 50, and 500 mu g/100 g). Four days after treatment, PHBA produced a dose-dependent response on vaginal cornification and uterotrophic activity in both immature and adult ovariectomized mice. The relative uterotrophic potency of PHBA (500 mu g/100 g) to E-2 (1 mu g/100 g) was 0.0011 in immature mice and 0.0018 in ovariectomized animals. (C) 1997 Academic Press.