1999 Fiscal Year Final Research Report Summary
Development of analytical methods for DNA-carcinogen adducts
| Project/Area Number |
10044290
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | OKAYAMA UNIVERSITY |
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Principal Investigator |
KOBAYASHI Sakae (有元 佐賀惠) Okayama university, Faculty of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (90212654)
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| Co-Investigator(Kenkyū-buntansha) |
NEGISHI Tomoe Okayama university, Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助教授 (80116491)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Carcinogens / DNA adducts / Highly sensitive analysis / Instrumental analysis / Heterocyclic amine / Alkylative agent |
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| Research Abstract |
We investigated the quantitative analysis of IQ adducts using liquid chromatography-mass-spectrometry (LCMS). The synthesis of the adduct N-(deoxyguanosin-8-yl)-IQ (C8-IQdG) was achieved by a method modified from Snyderwine et al (1988). LCMS was performed on a Quattro BQ tandem quadrupole mass spectrometer coupled to an HPLC system fitted with a Hypersil BDS C18 column. The retention time of C8-IQdG was 7.8 min, using an isocratic separation. Multiple Reaction Monitoring was used to detect the ion of C8-IQdG (m/z 464.6) and the product ion m/z 348.6 The detection limit of C8-IQdG using was 2.5 〜10 fmol / injection, in contrast that of C8-IQdG by UV-absorption at 342 nm was 10 pmol/injection. The amounts of C8-IQdG in DNA samples can be detectable in this method. We also investigate the quantitative analysis of alkylguanine in calf thymus DNA treated with a nitrosamine plus ultra-violet irradiation (UNA) using LCMS. We observed the formation of 【O!-】ィイD16ィエD1-methylguanine (OィイD16ィエD1-mdG), 【N!-】-methylguanine (NィイD17ィエD1-meG) and 7, 8-dihydro-8-oxodeoxygaunine (8-oxodG) in calf thymus DNA treated with NDMA plus UVA. We also observed mutagenicity of NDMA under irradiation of natural sunlight in 【S!-】.typhimurium. Furthermore, we detected the formation of OィイD16ィエD1-meG, NィイD17ィエD1-meG and 8-oxodG in calf thymus DNA treated with NDMA plus simulated sunlight. Regarding the mutagenesis of 【S!-】.typhimurium by NDMA plus UVA, we have now identified and quantified OィイD16ィエD1-meG formed in the genomic DNA of the bacteria under conditions of the mutagenesis.
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[Publications] Watanabe, T., Mazumder, T. K. Yamamoto, A., Nagai, S., Arimoto-Kobayashi, S., Hayatsu, H. and Terabe, S.: "A simple and rapid method for analysis the Monascuc pigment-mediated degradation of mutagenic 3-hydroxyamino-1-methyl-5H-pyrido[4.3-b] indole (Trp-P2(NHOH)) by incapillary micellar electrokinetic chromatography"Mutation Research. 444. 75-83 (1999)
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