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Monitoring of dimet...
Monitoring of dimethyl sulphate-induced N3-methyladenine, N7-methylguanine and O6-methylguanine DNA adducts using reversed-phase high performance liquid chromatography and mass spectrometry
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Chadt, J (författare)
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Sykora, D (författare)
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- Nilsson, Robert (författare)
- Stockholms universitet,Institutionen för genetik, mikrobiologi och toxikologi
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(creator_code:org_t)
- Elsevier BV, 2008
- 2008
- Engelska.
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Ingår i: Journal of chromatography. B. - : Elsevier BV. - 1570-0232 .- 1873-376X. ; 867, s. 43-48
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- 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.
Nyckelord
- N3-methyladenine; N7-methylguanine; O6-methylguanine; DNA methylated adducts; Dimethyl sulphate; HPLC; LC–MS–ESI(+)
- NATURAL SCIENCES
- NATURVETENSKAP
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- art (ämneskategori)
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