ABF, Analytisch-Biologisches Forschungslabor GmbH, Semmelweisstr. 5, Planegg 82152, Germany.
Chem Res Toxicol. 2024 Nov 18;37(11):1884-1902. doi: 10.1021/acs.chemrestox.4c00258. Epub 2024 Oct 15.
Tobacco smoke contains several electrophilic constituents which are capable of forming adducts with nucleophilic sites in DNA and proteins like hemoglobin (Hb) and albumin. New nicotine and tobacco products are discussed as less harmful forms of tobacco use compared to smoking combustible cigarettes (CC) due to reduced exposure to harmful constituents. Hence, the adduct profile in users of various tobacco/nicotine products is expected to differ characteristically. In this article, we present a novel nontargeted screening strategy using GC-MS/MS for Hb adducts based on the analysis of the respective derivatized N-terminal valine adducts after modified Edman degradation. We analyzed blood samples from a clinical study with habitual users of CCs, electronic cigarettes, heated tobacco products (HTPs), oral tobacco, nicotine replacement therapy products and nonusers of any tobacco/nicotine products. Our nontargeted approach revealed significant differences in the Hb adduct profiles of the investigated tobacco/nicotine product user groups. Adduct identification was performed by means of an internal database, retention time estimations based on the theoretical boiling points, as well as in-house synthesized reference compounds. Several chemicals that form adducts with Hb could be identified: methylating and ethylating agents, ethylene oxide, acrylonitrile, acrylamide, glycidamide and 4-hydroxybenzaldehyde. Levels were elevated in smokers compared to all other groups for Hb adducts from methylating agents, ethylene oxide, acrylonitrile, acrylamide and glycidamide. Our approach revealed higher concentrations of Hb adducts formed by ethylation, acrylamide and glycidamide in users of HTPs compared to nonusers. However, concentrations for the latter two were still lower than in smokers. Due to their long half-lives, Hb adducts related to acrylonitrile, acrylamide (glycidamide), and ethylene oxide exposure may be useful for the biochemical verification of subjects̀ compliance in longitudinal and cross-sectional studies with respect to smoking and HTP use/abstinence.
烟草烟雾中含有几种亲电成分，能够与 DNA 和蛋白质（如血红蛋白 (Hb) 和白蛋白）中的亲核位点形成加合物。与可燃香烟 (CC) 相比，新型尼古丁和烟草产品被认为是危害较小的烟草使用形式，因为它们接触到的有害成分较少。因此，各种烟草/尼古丁产品使用者的加合物谱预计会有特征性的差异。在本文中，我们提出了一种使用 GC-MS/MS 分析血红蛋白加合物的新型非靶向筛选策略，该策略基于对经过改良的 Edman 降解后各自衍生的 N-末端缬氨酸加合物进行分析。我们分析了来自习惯性使用 CC、电子烟、加热烟草产品 (HTP)、口含烟草、尼古丁替代疗法产品和不使用任何烟草/尼古丁产品的临床研究的血液样本。我们的非靶向方法揭示了所研究的烟草/尼古丁产品使用者群体的血红蛋白加合物谱存在显著差异。通过内部数据库、基于理论沸点的保留时间估算以及内部合成的参考化合物对加合物进行鉴定。可以鉴定出几种与 Hb 形成加合物的化学物质：甲基化和乙基化试剂、环氧乙烷、丙烯腈、丙烯酰胺、缩水甘油酰胺和 4-羟基苯甲醛。与所有其他组相比，吸烟者的甲基化试剂、环氧乙烷、丙烯腈、丙烯酰胺和缩水甘油酰胺形成的 Hb 加合物水平升高。与非使用者相比，HTP 使用者的 Hb 加合物中乙基化、丙烯酰胺和缩水甘油酰胺的浓度更高。然而，后两者的浓度仍低于吸烟者。由于其半衰期较长，与丙烯腈、丙烯酰胺（缩水甘油酰胺）和环氧乙烷暴露相关的 Hb 加合物可能有助于在纵向和横断面研究中对吸烟和 HTP 使用/戒断的生物化学验证。
