Reactivity of the\n2‑Methylfuran Phase I Metabolite\n3‑Acetylacrolein Toward DNA

2-Methylfuran (2-MF) is a well-known industrial chemical\nand also\nformed via thermal treatment of food. One main source of 2-MF in the\nhuman diet is coffee. 2-MF is known to form 3-acetylacrolein (AcA,\n4-oxopent-2-enal) via cytochrome P 450 metabolism and further reacts\nwith amino acids in vivo. Still the reactivity toward other biomolecules\nis rather scarce. Therefore, AcA was synthesized, and its reaction\nwith 2′-deoxyadenosine (dA), 2′deoxyguanosine (dG),\n2′deoxycytosine (dC), and 2′-deoxythymidine (dT) was\ntested. For this purpose, adduct formation was performed by acid hydrolysis\nof 2,5-dihydro-2,5-dimethoxy-2-methylfuran (DHDMMF) as well as pure\nAcA. The structures of these adducts were confirmed by UPLC-ESI<sup>+</sup>-MS/MS fragmentation patterns and <sup>1</sup>H-/<sup>13</sup>CNMR spectra. Except for dT, which showed no reactivity, all adducts\nof AcA were characterized, which enabled the development of sensitive\nquantification methods via (U)­HPLC-ESI<sup>±</sup>-MS/MS. Pure\nAcA was synthesized by oxidation of 2-MF using dimethyldioxirane (DMDO),\nand its behavior in aqueous medium was studied. Incubations of AcA\nand isolated DNA of primary rat hepatocytes (pRH) showed time- and\ndose-dependent formation of the identified DNA adducts dA-AcA, dG-AcA,\nor dC-AcA. In contrast, the DNA adducts dA-AcA, dG-AcA, or dC-AcA\nwere not detected on a cellular level when pRH were incubated with\n2-MF or AcA. This indicates an efficient detoxification or reaction\nwith biomolecules in the cell, although the induction of other DNA\ndamage, possibly also by other metabolites, cannot be ruled out in\nprinciple.