As a first step in investigating the genotoxic effects of the principal metabolites of 1,3-butadiene (BD) in both rats and mice, splenocytes (which have little mixed function oxidase activity) from each specimen were exposed to a series of concentrations of either 3,4-epoxy-1-butene (EB) (20 to 931 μM) or 1,2:3,4-diepoxybutane (DEB) (2.5 to 160 μM) for 1 h. The splenocytes were then washed, cultured, and stimulated to divide with concanavalin A, and metaphases were analyzed for the induction of sister chromatid exchanges (SCEs) and chromosome aberrations (CAs). In addition, cells from some experiments were taken after exposure but before culture, and subjected to the single cell gel (SCG) assay to measure DNA damage in the form of DNA strand breakage and/or alkaline-labile sites. Initial studies indicate that EB does not induce cytogenetic damage in either rat or mouse G 0 splenocytes. However, DEB was an extremely potent SCE- and CA-inducer in both species with no species differences apparent. Neither DEB nor EB produced any statistically significant DNA-damaging effects as measured by the SCG assay.