Quantification of 1,N6-etheno-2′-deoxyadenosine in human urine by column-switching LC/APCI-MS/MS
Quantification of 1,N6-etheno-2′-deoxyadenosine in human urine by column-switching LC/APCI-MS/MS
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DOI：
10.1016/j.freeradbiomed.2004.02.068
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发表时间：
2004-06-01
影响因子：
7.4
通讯作者：
Poulsen, HE
中科院分区：
文献类型：
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作者：
Hillestrom, PR;Hoberg, AM;Poulsen, HE
1，N-6-亚乙基-2‘-脱氧腺苷(EpsilondA)是细胞氧化代谢过程中产生的几种致突变DNA修饰之一。据认为，这些背景DNA损伤可能导致各种疾病，如癌症。因此，人体对尿液中epsilondA的生物监测可能成为氧化应激相关DNA损伤的潜在标志物。现有的尿液EDA定量方法使用P-32后标记。我们建立了一种基于柱切换-大气压化学电离串联质谱仪(LC/APCIMS/MS)的无放射性、快速、简便的检测方法。柱温的差异被用来影响分析物保留和样品聚焦。在多反应监测(MRM)模式下，从3毫升尿液开始，提供的检测下限约为0.7 PM。28名非吸烟者和5名吸烟者的尿EDA排泄率为10.0~99.6pmol/24小时，与体重、年龄和血浆维生素C浓度无关。5名吸烟者每24小时排泄量为30.5+/-8.5，28名不吸烟者每24小时排出量为38.6+/-2.4pmolepsilondA，p=0.37(平均扫描电子显微镜)。所展示的性能水平表明，这种方法未来将适用于癌症和其他与人类氧化应激有关的疾病的研究。(C)2004 Elsevier Inc.保留所有权利。
1,N-6-Etheno-2'-deoxyadenosine (epsilondA) is one of several promutagenic DNA modifications arising from cellular oxidative metabolism. It is believed that these background DNA lesions may contribute to various diseases, such as cancer. Therefore, human biomonitoring of epsilondA in urine could be a potential marker for oxidative stress-related DNA damage. Existing methods for quantifying urinary edA use P-32 postlabeling. We have developed a nonradioactive, fast, and easier method based on column-switching liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry (LC/APCI-MS/MS) in the positive mode. Differences in column temperatures were used to influence analyte retention and sample focusing. With multiple reaction monitoring (MRM) mode the afforded limit of detection was about 0.7 pM when starting with 3 ml of urine. The urinary excretion rates of edA from 28 nonsmoking and 5 smoking men were 10.0-99.6 pmol/24 h, and did not correlate with body weight, age, or plasma vitamin C concentration. The 5 smokers excreted 30.5 +/- 8.5 and the 28 nonsmokers excreted 38.6 +/- 2.4 pmol epsilondA per 24 h, p = .37 (mean SEM). The demonstrated level of performance suggests the future applicability of this method to studies of cancer and other diseases related to oxidative stress in humans. (C) 2004 Elsevier Inc. All rights reserved.