Linkage of In Vitro Assay Results With In Vivo End Points, Phases 1 and 2

Our results clearly show that it is possible to establish linkages between the screening level in in vitro assays and higher order endpoints in fish that are influenced by estrogen, such as growth and gonadal sex differentiation (Table 1). As expected, the in vitro responses are the most sensitive endpoints tested and using them as a monitoring tool can provide a margin of safety for aquatic organisms that may be more sensitive to estrogenic chemicals than Menidia. Moreover, the margin of safety provided, represented by the ratio of in vivo to in vitro EC50s ranged from 0.9 to 9 for the potent estrogen E2 to 1.2 for E1 and 2.4 to 3.4 for NP (Table 1). There were no significant effects on growth, but merely trends, by any of the chemicals tested, suggesting that a higher “n” may be required to see appropriate effects on this endpoint. Using more concentrations for in vivo tests can refine these ratios.