In vitro prenatal developmental toxicity induced by some petroleum substances is mediated by their 3- to 7-ring PAH constituent with a potential role for the aryl hydrocarbon receptor (AhR).

•This study evaluates the in vitro developmental toxicity potency of a series of petroleum substances extracts using alternative testing strategies.•In vitro PDT potency of petroleum substances extracts, as quantified in the EST, is mediated by their 3-7 ring PAHs with a potential role for the AhR.•A good correlation exists when comparing the in-vitro with the in-vivo PDT potencies of the petroleum substances extracts under study.•Our findings support the hypothesis that 3- to 7- ring PAHs are the primary inducers of the PDT in some petroleum substances.•The EST could also be applied for in vitro PDT testing of other UVCBs.