Development of the adductome approach to detect DNA damage in humans
Development of the adductome approach to detect DNA damage in humans
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DOI：
10.1089/ars.2006.8.993
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发表时间：
2006-05-01
影响因子：
6.6
通讯作者：
Matsuda, Tomonari
中科院分区：
文献类型：
--
作者：
Kanaly, Robert A.;Hanaoka, Tomoyuki;Matsuda, Tomonari
开发旨在检测氧化应激和其他手段引起的DNA损伤的新策略，可能会促进我们对此类损伤在癌症病因学、衰老过程中的作用以及作为暴露的生物标志物的理解。本文报道了一种DNA加合物检测方法，其使用液相色谱-电喷雾电离串联质谱（LC/ESI-MS/MS）来检测人肺组织中的多种DNA加合物。该内收体分析策略被设计为在多反应离子监测模式（MRM）中检测正离子化的2 '-脱氧核苷加合物中2'-脱氧核糖的中性损失，所述多反应离子监测模式在质量范围从m/z 228.8至m/z 602.8中在总共374个跃迁中传输[M + H]（+）> [M + H - 116]（+）跃迁。数据分析经过优化，并结合全面的手动筛选过程，旨在最大限度地减少最终分析中出现的人为加合物的数量。在最后的分析中，将推定的加合物组织成内收体图谱，并通过稳定同位素稀释和与真实标准品的比较来明确确认所选的氧化加合物。讨论了该方法的应用前景。
The development of new strategies designed to detect DNA damage caused by oxidative stress and other means may advance our understanding of the roles of such types of damage in the etiology of cancers, in aging processes, and as biomarkers; of exposure. A DNA adduct detection method that uses liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) to detect multiple DNA adducts in human lung tissue is reported herein. This adductome analysis strategy is designed to detect the neutral loss of 2'-deoxyribose from positively ionized 2'-deoxynucleoside adducts in multiple reaction ion monitoring mode (MRM) transmitting the [M + H](+) > [M + H - 116](+) transition over a total of 374 transitions in the mass range from m/z 228.8 to m/z 602.8. Data analysis is optimized and coupled with a comprehensive manual screening process designed to minimize the number of artifactual adducts appearing in the final analysis. In the final analysis, putative adducts were organized into an adductome map and unambiguous confirmation of selected oxidative adducts were made by stable isotope dilution and comparison to authentic standards. The future applications of this method are discussed.