乙烯基DNA加合物检测技术研究

 为了深入的研究乙烯基DNA加合物,本文综述了DNA氧化损伤的种类、修复机制和产物以及3种乙烯基DNA加合物：乙烯基腺嘌呤(1,N⁶-ethenoadenine,εAde)、乙烯基鸟嘌呤(1,N²-ethenoguanine,εGua)和乙烯基胞嘧啶(3,N⁴-ethenocytosine,εCyt)的体内生成机制。讨论了目前检测乙烯基DNA加合物的方法,如³²P-标记法,气相色谱-质谱法,液相色谱-串联质谱法等。³²P-标记法在检测灵敏度方面表现较优；气相色谱-质谱法分析样品需要衍生化,因此易导致样品在前处理过程中损失；液相色谱-串联质谱法具有前处理方法简单、稳定性好、选择性强、灵敏度高等优点。乙烯基DNA加合物作为生物标志物对评价生物体脂质过氧化具有重要意义。液相色谱-串联质谱法是目前在检测乙烯基DNA加合物方面较为理想的方法。

Analytical Method for Etheno DNA Adducts

In order to deeply study etheno-DNA adducts, the variety of DNA oxidative damages, repair mechanisms, and their products are summarized. The generative mechanism in vivo and analytical method of etheno-DNA adducts, including 1, N⁶-ethenoadenine (εAde), 1, N²- ethenoguanine (εGua), and 3, N⁴-ethenocytosine (εCyt) are reviewed. The current analytical methods of etheno-DNA adducts have been discussed, which include Immunoaffinity Chromatography/³²P-postlabelling technique (IC-³²P), Gas Chromatogeraph-Mass Spectrometer (GC-MS). and Liquid Chromatography-tandem Mass Spectrometer (LC-MS/MS). IC-³²P has the excellent performance on the sensibility, but when determining etheno-DNA adducts, complex operations and many steps have been involved. As analytical devices, the characteristics of mass spectrometer make the device obtain ideal sensibility and specificity. GC-MS method has the lower limit of determination than that for LC-MS/MS method. As sample derivatization is needed, the sample usually cost more for the method of GC-MS during the pre-handling; LC-MS/MS method offers many practical advantages on the pre-handing, stability, selectivity, and sensibility. These advantages could enhance the efficiency of samples analysis. LC-MS/MS is the ideal analytical method for the research on etheno-DNA adducts. The mechanism of DNA oxidative damage is inconclusive. As the biomarkers of DNA oxidative damages, etheno-DNA adducts possess significant meanings on the risk evaluation of lipid peroxidation.