Simultaneous determination of O6-methyl-2′-deoxyguanosine, 8-oxo-7,8-dihydro-2′-deoxyguanosine, and 1,N6-etheno-2′-deoxyadenosine in DNA using on-line sample preparation by HPLC column switching coupled to ESI-MS/MS
Simultaneous determination of O6-methyl-2′-deoxyguanosine, 8-oxo-7,8-dihydro-2′-deoxyguanosine, and 1,N6-etheno-2′-deoxyadenosine in DNA using on-line sample preparation by HPLC column switching coupled to ESI-MS/MS
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DOI：
10.1016/j.jchromb.2005.10.046
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发表时间：
2006-01-18
影响因子：
3
通讯作者：
Lutz, WK
中科院分区：
文献类型：
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作者：
Brink, A;Lutz, U;Lutz, WK
O-6-甲基-2'-脱氧金苷 (O-6-mdGuo)、8-oxo-7,8-二氢-2'-脱氧鸟苷 (8-oxodGuo) 和 1,N-6-乙烯-2'-脱氧腺苷 (epsilon dAdo) 是源自内源性和外源性物质和作用（甲基化、羟基化、脂质过氧化产物）。开发了一种高度灵敏的定量方法，使用带有柱切换的液相色谱串联质谱法同时测量这些 DNA 加合物。合成氘代O-6-[H-2(3)]mdGuo并用作内标。 O-6-mdGuo、8-oxodGuo 和 epsilon dAdo 的定量限分别为 24、98 和 48 fmol。该方法显示三种加合物的线性范围分别为 0.24-125 pmol/ml、0.98-125 pmol/ml 和 0.49-62.5 pmol/ml。 O-6-mdGuo 线性浓度范围内的日间精度为 1.7% 至 9.3%，8-oxodGuo 为 10.6% 至 28.7%，edAdo 为 6.2% 至 10.4%。从未经治疗的 12 周龄雌性 F344 大鼠肝脏中分离出的 DNA 中，O-6-mdGuo 高于检测限（每 109 个正常核苷有 37 个加合物），但无法定量。 8-oxodGuo 和 edAdo 显示每 109 个正常核苷分别有 500 个和 130 个加合物的背景水平。大鼠口服 50 μg/kg b.wt 二甲基亚硝胺治疗 1 小时后进行 DNA 分析。不影响 8-oxodGuo 和 edAdo 的水平，但导致每 10(9) 个正常核苷产生 200 个 O-6-mdGuo 加合物。所开发的方法将用于研究外源 DNA 加合物作为背景 DNA 损伤增量的生物学意义以及调节因子（例如 DNA 修复）的作用。 (c) 2005 Elsevier B.V. 保留所有权利。
O-6-Methyl-2'-deoxyauanosine (O-6-mdGuo), 8-oxo-7,8-dihydro-2'-deoxyguano sine (8-oxodGuo), and 1,N-6-etheno-2'-deoxyadenosine (epsilon dAdo) are promutagenic DNA lesions originating from both endogenous and exogenous agents and actions (methylation, hydroxylation, lipid peroxidation products). A highly sensitive quantitative method was developed to measure these DNA adducts simultaneously, using liquid chromatography tandem mass spectrometry with column switching. Deuterated O-6-[H-2(3)]mdGuo was synthesized and used as internal standard. The limits of quantification for O-6-mdGuo, 8-oxodGuo, and epsilon dAdo were 24, 98, and 48 fmol on column, respectively. The method showed linearity in the range 0.24-125 pmol/ml, 0.98-125 pmol/ml, and 0.49-62.5 pmol/ml for the three adducts, respectively. The inter-day precision in the linear concentration range was between 1.7 and 9.3% for O-6-mdGuo, 10.6 and 28.7% for 8-oxodGuo, and 6.2 and 10.4%, for edAdo. In DNA isolated from liver of untreated 12-week-old female F344 rats, O-6-mdGuo was above the limit of detection (37 adducts per 109 normal nucleosides) but could not be quantified. 8-oxodGuo and edAdo showed background levels of 500 and 130 adducts per 109 normal nucleosides, respectively. DNA analyzed 1 h after treatment of rats with dimethylnitrosamine by oral gavage of 50 mu g/kg b.wt. did not affect the levels of 8-oxodGuo and edAdo but resulted in 200 O-6-mdGuo adducts per 10(9) normal nucleosides. The method developed will be of use to study the biological significance of exogenous DNA adducts as an increment to background DNA damage and the role of modulating factors, such as DNA repair. (c) 2005 Elsevier B.V. All rights reserved.