A unique terrestrial large impact melt sheet is preserved in the 1850 Ma Sudbury Structure, Ontario. We have undertaken a Pb isotope investigation of the southern limb of the melt sheet, termed the South Range Main Mass. The model initial Pb isotope ratios (207Pb/204Pbm) vary stratigraphically through the predominantly quartz monzogabbroic Lower Unit, varying from 15.40 to 15.45 at the base to ca. 15.35 at the top of the sequence. Lateral variations of similar range occur in basal Lower Unit samples over scales of less than 5 km. The range of these variations is similar to those of locally exposed upper crustal target rocks, and it is evident that the melt sheet has efficiently preserved inherited variability. During the violent phases of crater formation superheated impact melts are expected to be well-mixed mechanically, therefore significant post-impact melting of target rocks, fallback material and entrained clasts is required to explain such heterogeneity. The Sudbury Structure hosts world class Ni-Cu-PGE sulphide ore deposits. Systematic variation in 207Pb/204Pbm occurs throughout sulphide ores within the Creighton Embayment, from massive (15.42–15.45) to interstitial (ca. 15.40–15.41) and disseminated (ca. 15.39) sulphide. Linking the Pb isotope composition of these ores to the immediately overlying Lower Unit stratigraphy, a protracted sulphide segregation history is apparent. Massive sulphides segregated early, prior to or during initial silicate crystallisation, although the total time involved in sulphide accumulation spanned much of the crystallisation of the Lower Unit. It is also shown that lateral variations in Ni depletion throughout the Main Mass correlate with Pb isotopes. Those segments with the strongest chalcophile element depletion signatures, reflecting the accumulation of significant basal sulphides, have high initial Pb isotope values, consistent with early sulphide segregation. The characterisation of Pb isotopic heterogeneity has therefore provided insights into the evolution and scales of mixing of the melt sheet, with the identified chemical variability between melt cells having a significant influence on ore forming processes.