Determination of DNA adducts by combining acid-catalyzed hydrolysis and chromatographic analysis of the carcinogen-modified nucleobases
| العنوان: | Determination of DNA adducts by combining acid-catalyzed hydrolysis and chromatographic analysis of the carcinogen-modified nucleobases |
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| المؤلفون: | Elvis M. K. Leung, Kailin Deng, Tin-Yan Wong, Wan Chan |
| المصدر: | Analytical and Bioanalytical Chemistry. 408:953-961 |
| بيانات النشر: | Springer Science and Business Media LLC, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | 0301 basic medicine, Chromatography, Hydrolysis, Biochemistry, Analytical Chemistry, Adduct, Nucleobase, DNA Adducts, 03 medical and health sciences, chemistry.chemical_compound, 030104 developmental biology, chemistry, Deoxyadenosine, DNA adduct, Carcinogens, Deoxyguanosine, Depurination, Acids, Chromatography, High Pressure Liquid, DNA |
| الوصف: | The commonly used method of analyzing carcinogen-induced DNA adducts involves the hydrolysis of carcinogen-modified DNA samples by using a mixture of enzymes, followed by (32)P-postlabeling or liquid chromatography (LC)-based analyses of carcinogen-modified mononucleotides/nucleosides. In the present study, we report the development and application of a new approach to DNA adduct analysis by combining the H(+)/heat-catalyzed release of carcinogen-modified nucleobases and the use of LC-based methods to analyze DNA adducts. Results showed that heating the carcinogen-modified DNA samples at 70 °C for an extended period of 4 to 6 h in the presence of 0.05% HCl can efficiently induce DNA depurination, releasing the intact carcinogen-modified nucleobases for LC analyses. After optimizing the hydrolysis conditions, DNA samples with C8- and N (2) -modified 2'-deoxyguanosine, as well as N (6) -modified 2'-deoxyadenosine, were synthesized by reacting DNA with 1-nitropyrene, acetaldehyde, and aristolochic acids, respectively. These samples were then hydrolyzed, and the released nucleobase adducts were analyzed using LC-based analytical methods. Analysis results demonstrated a dose-dependent release of target DNA adducts from carcinogen-modified DNA samples, indicating that the developed H(+)/heat-catalyzed hydrolysis method was quantitative. Comparative studies with enzymatic digestion method on carcinogen-modified DNA samples revealed that the two hydrolysis methods did not yield systematically different results. |
| تدمد: | 1618-2650 1618-2642 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::60b8e8548f210bef610d5f2637baad97 https://doi.org/10.1007/s00216-015-9186-0 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....60b8e8548f210bef610d5f2637baad97 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 16182650 16182642 |
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