アセトアルデヒドを由来とするDNA損傷の分析法に関する研究

概要

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Acetaldehyde reacts with exocyclic amino groups of guanine in DNA to form N^2-ethylffuanine (N^2-Et-Gua) and cyclic 1,N^2-propanoguanine adduct (CPr-Gua). Until now, the ^<32>P-postlabeling assay is one of the most useful methods for the detection of DNA adducts. However, it is not a safe one because it requires radioactive compounds. In this study, more safe and precise methods for the detection of DNA adducts using capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC) were developed. Using CE, we succeeded to develop a separation system that enabled us to control the migration time of nucleotides by changing the pH of the running buffer solution and the concentrations of buffer additives. Also, we developed an electrochemical detection cell for CE, which carried out the selective detection of normal and damaged guanines. Using liquid chromatography-mass spectrometry (LC-MS), a highly sensitive detection of DNA adducts was achieved, and it could be used to analyze real DNA samples. We considered about damage in DNA through the LC-MS analysis of DNA of cultured human cells and calf thymus DNA that were exposed to acetaldehyde. The developed detection methods are expected to replace the ^<32>P-postlabeling assay, or to be complementary to the assay. These results will contribute for DNA adducts studies in connection with carcinogenesis.
社団法人日本分析化学会の論文
2004-10-05