一种用于快速检测和定量 DNA 加合物 dG-ABP 的差分迁移谱/质谱平台。
A differential mobility spectrometry/mass spectrometry platform for the rapid detection and quantitation of DNA adduct dG-ABP.
机构信息
Northeastern University, Department of Chemistry and Chemical Biology and Barnett Institute, Boston, MA 02115, USA.
出版信息
Rapid Commun Mass Spectrom. 2013 Jul 15;27(13):1473-80. doi: 10.1002/rcm.6591.
Abstract
RATIONALE
There is continued interest in exploring new analytical technologies for the detection and quantitation of DNA adducts, biomarkers which provide direct evidence of exposure and genetic damage in cells. With the goal of reducing clean-up steps and improving sample throughput, a Differential Mobility Spectrometry/Mass Spectrometry (DMS/MS) platform has been introduced for adduct analysis.
METHODS
A DMS/MS platform has been utilized for the analysis of dG-ABP, the deoxyguanosine adduct of the bladder carcinogen 4-aminobiphenyl (4-ABP). After optimization of the DMS parameters, each sample was analyzed in just 30 s following a simple protein precipitation step of the digested DNA.
RESULTS
A detection limit of one modification in 10^6 nucleosides has been achieved using only 2 µg of DNA. A brief comparison (quantitative and qualitative) with liquid chromatography/mass spectrometry is also presented highlighting the advantages of using the DMS/MS method as a high-throughput platform.
CONCLUSIONS
The data presented demonstrate the successful application of a DMS/MS/MS platform for the rapid quantitation of DNA adducts using, as a model analyte, the deoxyguanosine adduct of the bladder carcinogen 4-aminobiphenyl.
摘要
原理
人们一直有兴趣探索用于检测和定量 DNA 加合物的新分析技术，这些生物标志物为细胞中的暴露和遗传损伤提供了直接证据。为了减少清洗步骤并提高样品通量，引入了用于加合物分析的差分迁移率光谱/质谱（DMS/MS）平台。
方法
已经利用 DMS/MS 平台分析了脱氧鸟苷-ABP，即膀胱致癌物 4-氨基联苯（4-ABP）的脱氧鸟苷加合物。在优化 DMS 参数后，仅需 2 µg DNA 进行简单的蛋白质沉淀步骤，每个样品在 30 秒内即可进行分析。
结果
使用仅 2 µg DNA 即可实现 10^6 个核碱基中一个修饰的检测极限。还进行了液相色谱/质谱的简短比较（定量和定性），突出了使用 DMS/MS 方法作为高通量平台的优势。
结论
所提供的数据表明，成功应用 DMS/MS/MS 平台快速定量 DNA 加合物，以膀胱致癌物 4-氨基联苯的脱氧鸟苷加合物作为模型分析物。
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