| 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 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)
TRACEY B. MRC Toxicology Chit, Biomonitoring and Mo, 上級研究員(研究職)
SHUKER D.E.G MRC Toxicology Chit, Biomonitoring and Mo, 上級研究員
FARMER P.B. MRC Toxicology Chit, Biomonitoring and Mo, 教授
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1998: ¥2,800,000 (Direct Cost: ¥2,800,000)
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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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