The late Eocene was marked by multiple impact events, possibly related to a comet or asteroid shower. Marine sediments worldwide contain evidence for at least two closely spaced impactoclastic layers. The upper layer might be correlated with the North American tektite-strewn field (with the 85-km-diameter Chesapeake Bay impact structure [USA] as its source crater), although this is debated, whereas the lower, microkrystite layer (with clinopyroxene [cpx]-bearing spherules) was most likely derived from the 100-km-diameter Popigai impact crater (Russia). The Eocene-Oligocene global stratotype section and point is located at Massignano, Italy, and below the boundary, in the late Eocene, at the 5.61 m level, shocked quartz and pancake-shaped smectite spherules that contain (Ni- and Cr-rich) magnesioferrite spinel crystals are found. These are associated with a positive Ir anomaly in deposits with the same age as the Popigai-derived cpx spherule layer. This layer is overlain by another Ir-rich layer, likely due to another large impact event. From a large amount of “pancake-bearing” rock, we isolated a few hundred milligrams of this spinel-rich material. The tungsten isotopic composition of this material shows more or less a terrestrial composition. However, the spinel-rich materials have excess ⁵⁴Cr values (expressed as ε⁵⁴Cr, which is the per ten thousand deviation of the ⁵⁴Cr/⁵²Cr ratio from a terrestrial standard) of around –0.4 to –0.5 ε⁵⁴Cr, which distinctly point to an ordinary chondritic impactor. This result supports the asteroid impact interpretation but not the comet impact hypothesis.