The Chesapeake Bay impact structure, which is 85 km in diameter and 35.5 Ma old, was drilled and cored in a joint International Continental Scientific Drilling Program (ICDP) and U.S. Geological Survey (USGS) drilling project at Eyreville Farm, Virginia, U.S.A. In the Eyreville drill core, 154 m of impact breccia were recovered from the depth interval 1397–1551 m. Major- and trace-element concentrations were determined in 75 polymict impactite samples, 10 samples of cataclastic gneiss blocks, and 24 clasts from impactites. The chemical composition of the polymict impactites does not vary much in the upper part of the section (above ~1450 m), whereas in the lower part, larger differences occur. Polymict impactites show a decrease of SiO₂ content, and slight increases of TiO₂, Al₂O₃, and Fe₂O₃ abundances, with depth. This is in agreement with an increase of the schist/gneiss component with depth. Concentrations of siderophile elements (Co, Ni) are lower in the polymict impactites than in the basement-derived schists and do not indicate the presence of an extraterrestrial component. The five petrographically determined types of melt particles, i.e., clear glass, altered melt, recrystallized silica melt, melt with microlites, and dark-brown melt, have distinct chemical compositions. Mixing calculations of the proportions of rocks involved in the formation of various polymict impactites and melt particles were carried out using the Harmonic least-squares MiXing (HMX) calculation program. The calculations suggest that the metamorphic basement rocks (i.e., gneiss and schist) constitute the main component of the polymict impactites, together with significant sedimentary and possible minor pegmatite/granite and amphibolite components. The sedimentary component is derived mostly from a sediment characterized by a composition similar to that of the Cretaceous Potomac Formation. Compositions of the melt particles were modeled as mixtures of target rocks or major rock-forming minerals. However, the results of the mixing calculations for the melt particles are not satisfactory, and the composition of the particles could have been modified by hydrothermal alteration. Carbon isotope ratios were determined for 18 samples. The results imply a hydrothermal origin for the carbonate veins from the basement-derived core section; carbon-rich sedimentary clasts from the Exmore breccia and suevite have a δ¹³C range typical for organic matter in sediments.