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Human experimentation was undertaken to determine whether a mixture of vapourized diacetyl and 2,3-pentanedione (Ac-2,3-PnD) is inhaled at a greater rate and at a greater concentration than is the sum of vapours of the two compounds. In this crossover trial, participants were randomized to receive a 40-min breathing exposure to either Ac-2,3-PnD or diacetyl plus PnD. The mixture produced greater particle concentrations and respiratory complaints than either compound alone. The interval between the first inhalation of each substance was chosen to achieve a similar concentration of vapours of each compound. Although the single compounds did not produce any detectable pattern of oxidized DNA adducts in the peripheral blood mononuclear cells, the mixture produced a similar level of dG-C8-PnD and dG-C8-Ac adducts, but a smaller level of dG-C8-Ac-2,3-PnD, dG-C8-Ac-PnD, dG-C8-Ac-PnD-2 and dG-C8-Ac-PnD-3 adducts. Thus, the mixture of diacetyl plus PnD appeared to be less toxic per unit of exposure, whereas the single compounds did not differ in their rate or magnitude of DNA damage. Although the potency of diacetyl for inducing oxidized DNA damage, previously documented in vitro, was verified in these human studies, the repeated-exposure concentrations used in this study produced greater amounts of inhaled diacetyl than previously tested, and were toxic to a greater extent, than the mixture of the two compounds.There’s