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Monitoring of dimethyl sulphate-induced N3-methyladenine, N7-methylguanine and O6-methylguanine DNA adducts using reversed-phase high performance liquid chromatography and mass spectrometry
Stockholm University, Faculty of Science, Department of Genetics, Microbiology and Toxicology.
2008 (English)In: Journal of chromatography. B, ISSN 1570-0232, Vol. 867, 43-48 p.Article in journal (Refereed) Published
Abstract [en]

This work describes the determination of N3-methyladenine, N7-methylguanine and O6-methylguanine adducts in dimethyl sulphate-treated salmon-testes DNA employing reversed-phase high performance liquid chromatography (RP-HPLC) with UV–vis detection, followed by mass-spectrometric verification using electrospray ionisation in positive mode ESI(+). Within validation parameters, accuracy, precision, calibration parameters, limit of detection (LOD) and quantitation (LOQ) as well as stability of standard stock solutions were tested and presented for UV/vis detection. The limit of detection (LOD) was found to be 0.1 ng/mL for N3-methyladenine and 0.2 ng/mL for both N7-methylguanine and O6-methylguanine (S/N = 3). The limit of quantitation (LOQ) was found to be 0.5 ng/mL for all measured compounds, (S/N = 10). Quantitative results were obtained for each substance based on eight-point calibration. Intra- and inter-day precisions were within 1.73–6.96 and 2.26–7.58%, respectively, and correlation coefficients of calibration curves (R2) ranged from 0.9992 to 0.9997. Relative proportion of N7-methylguanine was accounted for 61.53 ± 2.97% (R.S.D. = 4.8), N3-methyladenine for 38.19 ± 2.99% (R.S.D. = 9.6) and O6-methylguanine for 0.29 ± 0.02% (R.S.D. = 5.1), respectively. The application of the above-mentioned techniques provides a valuable contribution for simultaneous determination of methylated DNA adducts, and may represent a suitable approach for similar monitoring/screening studies.

Place, publisher, year, edition, pages
2008. Vol. 867, 43-48 p.
Keyword [en]
N3-methyladenine; N7-methylguanine; O6-methylguanine; DNA methylated adducts; Dimethyl sulphate; HPLC; LC–MS–ESI(+)
National Category
Natural Sciences
URN: urn:nbn:se:su:diva-31912DOI: 10.1016/j.jchromb.2008.03.008OAI: diva2:278988
Available from: 2009-12-01 Created: 2009-12-01 Last updated: 2009-12-03Bibliographically approved

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Nilsson, Robert
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