Department of Chemistry, Division of Food Chemistry and Toxicology, University of Kaiserslautern-Landau, Kaiserslautern D-67663, Germany.
J Agric Food Chem. 2024 Nov 13;72(45):25319-25329. doi: 10.1021/acs.jafc.4c07280. Epub 2024 Nov 4.
2-Methylfuran (2-MF) is a well-known industrial chemical and also formed via thermal treatment of food. One main source of 2-MF in the human diet is coffee. 2-MF is known to form 3-acetylacrolein (AcA, 4-oxopent-2-enal) via cytochrome P 450 metabolism and further reacts with amino acids in vivo. Still the reactivity toward other biomolecules is rather scarce. Therefore, AcA was synthesized, and its reaction with 2'-deoxyadenosine (dA), 2'deoxyguanosine (dG), 2'deoxycytosine (dC), and 2'-deoxythymidine (dT) was tested. For this purpose, adduct formation was performed by acid hydrolysis of 2,5-dihydro-2,5-dimethoxy-2-methylfuran (DHDMMF) as well as pure AcA. The structures of these adducts were confirmed by UPLC-ESI-MS/MS fragmentation patterns and H-/CNMR spectra. Except for dT, which showed no reactivity, all adducts of AcA were characterized, which enabled the development of sensitive quantification methods via (U)HPLC-ESI-MS/MS. Pure AcA was synthesized by oxidation of 2-MF using dimethyldioxirane (DMDO), and its behavior in aqueous medium was studied. Incubations of AcA and isolated DNA of primary rat hepatocytes (pRH) showed time- and dose-dependent formation of the identified DNA adducts dA-AcA, dG-AcA, or dC-AcA. In contrast, the DNA adducts dA-AcA, dG-AcA, or dC-AcA were not detected on a cellular level when pRH were incubated with 2-MF or AcA. This indicates an efficient detoxification or reaction with biomolecules in the cell, although the induction of other DNA damage, possibly also by other metabolites, cannot be ruled out in principle.
2-甲基呋喃（2-MF）是一种知名的工业化学品，也可通过食品的热处理形成。2-MF 是人类饮食中 2-MF 的主要来源之一。2-MF 已知通过细胞色素 P450 代谢形成 3-乙酰丙烯醛（AcA，4-氧戊-2-烯醛），并进一步在体内与氨基酸反应。然而，其与其他生物分子的反应性仍然相当匮乏。因此，合成了 AcA，并测试了其与 2'-脱氧腺苷（dA）、2'-脱氧鸟苷（dG）、2'-脱氧胞苷（dC）和 2'-脱氧胸苷（dT）的反应。为此，通过酸水解 2,5-二氢-2,5-二甲氧基-2-甲基呋喃（DHDMMF）和纯 AcA 进行加合物形成。通过 UPLC-ESI-MS/MS 碎片模式和 H-/CNMR 光谱证实了这些加合物的结构。除了无反应性的 dT 外，所有 AcA 的加合物都得到了表征，这使得通过（U）HPLC-ESI-MS/MS 开发了灵敏的定量方法。纯 AcA 通过使用二甲氧基二环氧乙烷（DMDO）氧化 2-MF 合成，并研究了其在水介质中的行为。AcA 与原代大鼠肝细胞（pRH）的分离 DNA 的孵育显示出时间和剂量依赖性形成鉴定的 DNA 加合物 dA-AcA、dG-AcA 或 dC-AcA。相比之下，当 pRH 与 2-MF 或 AcA 孵育时，在细胞水平上未检测到 DNA 加合物 dA-AcA、dG-AcA 或 dC-AcA。这表明在细胞中存在有效的解毒或与生物分子的反应，尽管不能排除其他 DNA 损伤的诱导，可能也由其他代谢物引起。
