臺灣博碩士論文加值系統

1,N2-ethenoguanine (1,N2-eGua)是高度突變性的DNA傷害，它可由體內DNA與體內脂質過氧化形成的化學物質反應形成。其中1,N2-eGua 可經由人體酵素alkylpurine-DNA-N-glycosylase 從組織切除修復排至尿液。我們開發了一個高靈敏度和高特異性的同位素稀釋氣相層析負離子化學游離質譜法來分析人體尿液中1,N2-eGua。在樣品前處理部分，先使用強陽離子交換和C18-OH兩種固相萃取管柱濃縮樣品後，以親電性衍生化試劑（PFB-Br）衍生，再以Si固相萃取管柱純化樣品PFB2-1,N2-eGua，接著以氣相層析負離子化學游離質譜搭配選擇離子追蹤分析；並以結構相同但分子量較高的穩定同位素[13C1,15N2] 1,N2-eGua作為內標準物。此方法在0.5 mL人體尿液樣品中偵測到1,N2-eGua的存在，並準確地定量。本研究也提供了一個可信的方法來測量人體尿液中1,N2-eGua作為脂質過氧化和氧化壓力的生化指標。
無鹼基位置（apurinic/apyrimidinic sites）是體內鹼基切除修復機制切除由外來環境污染物和內生性化學物質所修飾之鹼基，但未接上正常鹼基前的過渡狀態。也會因硝化性修飾鹼基，在生理條件下醣苷鍵水解而硝化鹼基自發性的掉落而生成。我們在此描述兩個定量DNA中無鹼基位置的方法：螢光和表面電漿共振(LSPR)。以具專一性之胺氧官能基和DNA上無鹼基位置之開環去氧核醣醛類反應形成穩定的oxime ether，在含胺氧官能基之探針另一端接上螢光集團後以螢光定量分析。第二個方法為合成一端含胺氧基，另一端含硫醇的化合物。硫醇可與金奈米結合成奈米探針，當它與DNA上無鹼基位置之醛基反應成oxime ether，會有LSPR訊號增加產生。

1,N2-Ethenoguanine (1,N2-eGua) is promutagenic DNA lesions, known to derive from endogenous lipid peroxidation. Moreover, 1,N2-eGua is reported to be a primary substrate of human alkylpurine-DNA-N-glycosylase. Thus, this DNA lesion could be excised from tissue DNA and released into urine. In this study, a highly sensitive and quantitative assay based on isotope dilution gas chromatography/ negative ion chemical ionization/ mass spectrometry (GC/NICI/MS) was developed for analysis of 1,N2-eGua in human urine. The assay involved adduct enrichment by consecutive strong cation exchange and reversed phase solid phase extraction columns. The enriched sample was derivatized by pentafluorobenzyl bromide (PFB-Br) and PFB2-1,N2-eGua was analyzed by GC/NICI/MS after a Si SPE clean-up. The stable isotope of the analyte, [13C1,15N2] 1,N2-eGua, was used as internal standard for accurate quantification of urinary 1,N2-eGua in as little as 0.5 mL of human urine samples. This assay provides a valuable tool for measuring urinary 1,N2-eGua as a potential biomarker of DNA damage and repair derived form lipid peroxidation and oxidative stress in general.
Apurinic (AP) sites are important DNA repair intermediates in mutagenesis and carcinogenesis. These lesions are generated by hydrolysis of the N-glycosidic bonds. We have described two methods for quantification of DNA apurinic sites, i.e. fluorescence and localized surface plasma resonance (LSPR). Based on the specific reaction of the amionooxy group and the aldehyde moiety of the open ring deoxyribose of DNA apurinic sites, a stable oxime ether is formed. A fluorescence probe is synthesized with an aminooxy group at one end to react with the apurinic sites of DNA. Another approach is to synthesize a nano-probe with an aminooxy group on one end and thiol group on the other. The thiol is attached to gold nanoparticles. When it reacts with the aldehydic group of DNA apurinic sites, formation of the oxime ether should lead to signal enhancement in LSPR.

目錄
目錄 Ⅰ
圖表目錄 IV
附圖目錄                   V

第一章 序論 1
1-1 去氧核醣核酸(DNA)的簡介 1
1-2 DNA加成產物的種類 3
1-3 外環性乙烯基加成產物對生物體的影響 4
1-4 外環性乙烯基加成產物的來源 6
1-5 DNA加成產物的生化意義 12
1-6 DNA上無嘌呤位置的形成 15
1-7 負離子化學游離質譜法（negative ion chemical
ionization Mass Spectrometry, NICI/MS） 17
1-8 研究動機 18

第二章 實驗部分 22
2-1 儀器 22
2-2 材料 22
2-3 藥品 23
2-4 氣相層析儀系統條件 23
2-5 質譜儀系統條件 24
2-6 高效能液相層析儀（HPLC）的分析條件 24
2-7 乙烯基鳥糞嘌呤的合成 24
2-8 尿液中1,N2-乙烯基鳥糞嘌呤之分析 25
2-9 利用SCX固相萃取管柱收集0.5毫升人體尿液中的
1,N2-εGua之條件 26
2-10 利用C18-OH固相萃取管柱收集經由SCX純化之1,N2-εGua之條件 27
2-11 PFB衍生化反應的條件 27
2.-12 Si固相萃取PFB-εGua的條件 28
2-13 利用氣相層析儀搭配負離子化學游離質譜儀對人體尿液中1,N2- εGua含量偵測 29
2-14 1,N2- Gua定量曲線 29
2-15 螢光感測器與金奈米感測器的合成 29
2-16 DNA上無鹼基位置的形成 31
2-17 螢光感測的反應條件 32
2-18 螢光感測的定量曲線 32

第三章 實驗結果 34
3-1-1 合成1,N2-εGua的結果 34
3-1-2 以SCX和C18-OH SPE收集人體尿液中之1,N2-εGua方法之確立 35
3-1-3 以Si SPE收集PFB2-1,N2-eGua方法之最佳化 36
3-1-4 1,N2-εGua定量曲線的確立 36
3-1-5 利用GC/NICI/MS法量測人體尿液中1,N2-eGua含量 37
3-2-1 螢光感測器與金奈米感測器的合成結果 38
3-2-2 DNA上無鹼基位置的形成 40
3-2-3 螢光感測的最佳反應條件 41
3-2-4 螢光感測的定量曲線 41
3-2-5 螢光探針偵測AP-sites DNA  42

第四章 討論 43
第五章 結論 46
參考文獻 48
中英對照表 61
























圖表目錄
圖一. DNA上的鹼基結構和Watson-Crick之DNA中鹼基結構與配對方式 2
圖二.單股DNA之鏈狀結構              3
圖三. 外環乙烯基加成產物結構 5
圖四. vinyl chloride及ethyl carbamate在生物體內的代謝物與DNA之作用 7
圖五. HNE和DNA作用生成etheno adducts 8
圖六. 生物體內氧化壓力生成外環性DNA adducts 9
圖七. 維生素C誘導脂質過氧化產生DNA加成產物示意圖 11
圖八. 3’-乙醛磷酸及CAA和Ade作用，生成εAde的推測機制 11
圖九. 無鹼基位置上的半縮醛與開放形式的醛基與含有胺基親核反應機制 16
圖十. 乙烯基鳥糞嘌呤的合成機制圖 25
圖十一. 人體尿液中的1,N2- eGua 之分析流程圖 26
圖十二. PFB-εGua衍生化反應 28
圖十三. DNA上無鹼基位置分析流程圖 32
圖十四. 螢光探針的合成流程 39
圖十五. 奈米探針的合成流程 40
圖十六. 螢光探針和去氧核糖穩定的oxime ether產物 42
圖十七. 螢光探針偵測AP-sites DNA的線性關係 42
圖十八. 螢光探針偵測DAN螢光圖譜 44
表一. 尿液中1,N2-εGua的含量結果 (尿液樣品量為0.5 mL) 38
表二. 與NO2Cl反應之DNA中8NG和8NX之含量 41

附圖目錄
附圖一. 1,N2- eGua之HPLC-UV層析圖譜 64
附圖二. 1,N2- eGua以PFB-Br衍生化後的層析圖譜
   （a）為HPLC圖譜（b）為GC/MS圖譜 65
附圖三. 1,N2- eGua以PFB-Br衍生化後的層析圖譜HPLC圖譜 65
附圖四. 1,N2- eGua以PFB-Br衍生化後的GC/MS層析圖譜 66
附圖五. GC/NICI/MS分析1,N2- eGua以PFB-Br衍生化後的層析圖譜 67
附圖六. 10pg 1,5-PFB2-1,N2- eGua之GC/NICI/MS質譜圖 68
附圖七. 1,5-PFB2-1,N2- eGua 之GC/NICI/MS的LOD圖 69
附圖八. 1,N2- eGua 之GC/NICI/MS定量曲線 69
附圖九. GC/NICI/MS分析1,N2- eGua的控制組層析圖譜 70
附圖十. GC/NICI/MS分析吸煙者尿液樣品中1,N2- eGua的層析圖譜 71
附圖十一. GC/NICI/MS分析非吸煙者尿液樣品中1,N2- eGua的層析圖譜 72
附圖十二. 螢光探針3 400MHz NMR圖譜 73
附圖十三. 螢光探針3 ESI/MS圖譜 73
附圖十四. 金奈米探針10在有機相中表面電漿共振圖譜(a)為未接上9之金奈米粒子LSPR圖譜；(b)為接上9後之金奈米粒子10 LSPR圖 74
附圖十五. 金奈米探針10在水相中表面電漿共振圖譜(a)為未接上9之金奈米粒子LSPR圖譜；(b)為接上9後之金奈米粒子10 LSPR圖譜 74
附圖十六. pH值對以螢光法偵測AP sites的影響 75
附圖十七. 螢光感測之定量曲線(螢光強度為在激發波長323 nm，
   放射波長525 nm下偵測) 75

第六章 參考文獻
1.Fearon, E. R.; Vogelstein, B. (1990) A genetic model for colorectal tumorigenesis. Cell 61, 759-767.
2.Poirier, M. C.; Santella, R. M.; Weston, A. (2000) Carcinogen macromolecular adducts and their measurement. Carcinogenesis 21, 353-359.
3.Chen, H.-J.; Hong, C.-L. (2001) 內生的DNA加成產物之形成與其在體內含量之分析. Chemistry 59, 299-310.
4.Chen, H.-J. (2002) 質譜在分析DNA加成產物之重要性與其在毒物學上的意義. Chemistry 60, 189-209.
5.Weinberg, R.A. Oncogenes and the Molecular Origins of Cancer, Cold Spring Harbor Press, New York, 1989.
6.Hemminki, K.; Dipple, A.; Bartsch, H. DNA Adducts：Identification and Biological Significance, IARC, Lyon, 1994.
7. Hemminki, K. (1993) DNA adducts, mutations and cancer. Carcinogenesis 14, 2007-2012.
8. Miller, E. C.; Miller, J. A. (1966) Mechanisms of chemical carcinogenesis: nature of proximate carcinogens and interactions with macromolecules. Pharmacol. Rev. 18, 805-838.
9. Basu, A. K.; Wood, M. L.; Niedernhofer, L. A.; Essigmann, J. M. (1993) Mugatenic and genotoxic effects of three vinylchloride-induced DNA lesion : 1,N6-ethenoadenine, 3,N4-ethenocytosine, and 4-amino-5-(imidazol-2-yl)imidazole. Biochemistry 32, 12793-12801.
10.Cheng, K. C.; Preston, B. D.; Cahill, D. S.; Dosanjh, M. K.; Singer, B.; Loeb, L. A. (1991) The vinyl chloride DNA derivative N2,3-ethenoguanine produced G→A transitions in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 88, 9974-9978.
11. Chengt, K. C., Prestont, B. D., Cahillo, D. S., Dosanjh, M. K., Singer, B., Loebt, L. A., (1991) The vinyl chloride DNA derivative N2,3-ethenoguanine produces G - A transitions in Escherichia coli
Biochemistry, 88, 9974-9978.
12. Kronberg, L.; Sjöholm, R.; Karlsson, S. (1992) Formation of 3,N4-ethenocytidine, 1,N6-ethenoadenosine and 1,N2-ethenoguanosine in reactions of mucochloric acid with nucleosides. Chem. Res. Toxical. 5, 852-855.
13. Fernando, R. C.; Nair, J.; Barbin, A.; Miller, J. A.; Bartsch, H. (1996) Detection of 1,N6-ethenodeoxyadenosine and 3,N4-etheno¬-¬¬deoxy¬cytidine by immunoaffinity/ 32P-postlabelling in liver and lung DNA of mice treated with ethyl carbamate (urethane) or its metabolites. Carcinogenesis 17, 1711-1718.
14. Battaglia, R.; Conacher, H. B. S.; Page, B. D. (1990) Ethyl carbamate (urethane) in alcoholic beverages and foods: a review. Food Addit. Contam. 7, 477-496.
15. Ough, C. S. (1976) Ethyl carbamate in fermented beverages and foods. I. Naturally occurring ethyl carbamate. J. Agric. Food Chem. 24, 323-328.
16. Creech, J. L. Jr.; Johnson, M. N. (1974) Angiosarcoma of liver in the manufacture of polyvinyl chloride. J. Occup. Med. 16, 150-151.
17. Gedigk, P.; Muller, R.; Bechtelshelimer, H. (1975) Moephology of liver damage among polyvinyl chloride production workers. A report on 51 cases. Ann. N. Y. Acad. Sci. 246, 278-285.
18. Buffler, P. A.; Wood, S.; Eifler, C.; Suarez, L.; Kilian, D. J. (1979) Mortality experience of workers in a vinyl chloride monomer production plant. J. Occup. Med. 21, 195-203.
19. Guengerich, F. P. (1992) Roles of the vinyl chloride oxidation products 2-chlorooxiranes and 2-chloroacetaldehyde in the vitro formation of etheno adducts of nucleic acid bases. Chem. Res. Toxicol. 5, 2-5.
20. Zajdela, F.; Croisy, A.; Barbin, A.; Malaveille, C.; Tomatis, L.; Bartsch, H. M. (1980) Carcinogenicity of chloroethylene oxide, an ultimate reactive metabolite of vinyl chloride, and bis(chloromethyl)ether after subcutaneous administration and in initiation-promotion experiments in mice. Cancer Res. 40, 352-356.
21. Ames, B. N.; Gold, L. S. (1991) Endogenous mutagens and the causes of aging and cancer. Mutat. Res. 250, 3-16.
22. Lindahl, T. (1993) Instability and decay of the primary structure of DNA. Nature 362, 709-715.
23. Chung, F. L.; Chen, H. -J. C.; Nath, R. G. (1996) Lipid peroxidation as a potential endogenous source for the formation of exocyclic DNA adducts: a commentary. Carcinogenesis 17, 2105-2111.
24. Esterbauer, H.; Schaur, R. J.; Zollner, H. (1991) Chemistry and biochemistry of 4-hydroxynonenal, malondialdehyde and related aldehydes. Free Radic. Biol. Med. 11, 81-128.
25. Wu, H.-Y.; Lin, J.-K. (1995) Determination of aldehydic lipid peroxidation products with dabsylhydrazine by high-performance liquid chromatography. Anal. Chem. 67, 1603-1612.
26. Chen, H.-J. C.; Chung, F.-L. (1996) Epoxidation of trans-4-hydroxy-2-nonenal by fatty acid hydroperoxides and hydrogen peroxide. Chem. Res. Toxical. 9, 306-312.
27. Chen, H.-J. C.; and Chung, F.-L. (1994) Formation of etheno adducts in reactions of enals via autoxidation. Chem. Res. Toxical. 7, 857-860.
28. Chen, H.-J. C.; Gonzalez.; Shou, M.; Chung, F.-L. (1998) 2,3-Epoxy-4-hydroxynonanal, a potential lipid peroxidation product for etheno adduct formation, is not a substrate of human epoxide hydrolase. Carcinogenesis 19, 939-943.
29. Sodum, R. S.; Chung, F.-L. (1989) Structural characterization of adducts formed in the reaction of 2,3-epoxy-4-hydroxynonanal with deoxyguanosine. Chem. Res. Toxicol. 2, 23-28.
30. Sodum, R. S.; Chung, F.-L. (1991) Stereoselective formation of in vitro nucleic acid adducts by 2,3-epoxy-4-hydroxynonanal. Cancer Res. 51, 137-143.
31. Chung, F.-L.; Chen, H.-J. C.; Guttenplan, J. B.; Nishikawa, A.; Hard, G. C. (1993) 2,3-Epoxy-4-hydroxynonanal as a pontential tumor-initiating agent of lipid peroxidation. Carcinogenesis 14, 2073-2081.
32. Sodum, R. S.; Chung, F.-L. (1988) 1,N2-Ethenodeoxyguanosine as a potential marker for DNA adduct formation by trans-4-hydroxy-2-
nonenal. Cancer Res. 48, 320-326.
33. Nair, V.; Offerman, R. J. (1985) Ring-extended products from the reaction of epoxy carbonyl compounds and nucleic acid bases. J. Org. Chem. 50, 5627-5632.
34. Chen, H.-J. C.; Wu, C.-F.; Hong, C,-L.; Chang, C.-M. (2004) Urinary excretion of 3,N4-Etheno-2’-deoxycytidine in humans as a biomarker of oxidative stress : association with cigarette smoking. Chem. Res. Toxicol. 17, 896-903.
35. Nair, J.; Gal, A.; Tamir, S.; Tannenbaum, S. R.; Wogan, G.. N., Bartsch, H. (1998) Etheno adduct in spleen DNA of SJL mice stimulated to overproduce nitric oxide. Carcinogenesis 19, 2081-2084.
36. Djuric, Z.; Heilbrun, L. K.; Reading, B. A.; Boomer, A.; Valeriote, F. A.; Martino, S. (1991) Effect of a low-fat diet on levels oxidative damage to DNA to human peripheral nucleated blood cells. J. Natl. Cancer Inst. 83, 766-769.
37. Badwey, J. A.; Curnutte, J. T.; Robinson, J. M.; Berde, C. B.; Karnovsky, M. J.; Karnovsky, M. L. (1984) Effects of free fatty acids on release of superoxide and on change of shape by human neutrophils. J. Biol. Chem. 259, 7870-7877.
38. Chen, H.-J. C.; Gonzalez; Shou, M.; Chung,-F. L. (1996) 2,3-Epoxy-4-hydroxynonanal is not a substrate for human epoxide hydrolase. Carcinogenesis. 19, 939-943.
39. Laib, R. J. (1986) The role of cyclic base adducts in vinyl-chloride-induced carcinogenesis: studies on nucleic acid alkylation in vivo. In the role of cyclic nucleic acid adducts in carcinogenesis and mutagenesis. IARC Sci. Publ. 70, 101-108.
40. Bartsch, H.; Barbin, A.; Marion, M. J.; Nair, J.; Guichard, Y. (1994) Formation, detection and role in carcinogenesis of etheno bases in DNA. Drug Metab. Rev. 26, 349-356.
41. Leonard, N. J. (1992) Etheno-bridged nucleotides in structure diagnosis and carcinogenesis. Chemtracts Biochem. Mol. Biol. 3, 273-297.
42. Bartsch, H.; Montesano, R. (1975) Mutagenic and carcinogenic epoxide of vinyl chloride. Mutat. Res. 32, 93-114.
43. Proteggente, A. R.; Rehman, A.; Hallwell, B.; Rice-Evens, C. A. (2000) Potenetial problems of ascorbate and iron supplementation: pro-oxidant effect in vivo ? Biochem. Biophys. Res. Commun. 277, 535-541.
44. Lee, S. H.; Oe, Tomoyuki; Blair, I. A. (2001) Vitamin C-induced decomposition of lipid hydroperoxides to endogenous genotoxins. Science 15, 2083-2086.
45. Jones, W. R.; Dedon, P. C. (1999) DNA oxidation as a source of endogenous electrophiles: Formation of ethenoadenine adducts in γ-irradiated DNA. J. Am. Chem. Soc. 121, 9231-9232.
46. Nair, J.; Carmichael, P. L.; Fernando, R. C.; Phillips, D. H.; Strain, A. J.; Bartsch, H. (1998) Lipid peroxidation-induced etheno-DNA adducts in the liver of patients with the genetic metal storage disorders Wilson’s disease and primary hemochromatosis. Cancer Epidemiol. Biomarkers Prev. 7, 435-440.
47. Nair, J.; Vaca, C. E.; Velic, I.; Mutanen, M.; Valsta, L. M.; Bartsch, H. (1997) High dietary ω-6 polyunsaturated fatty acids drastically increase the formation of etheno-DNA base adducts in white blood cell of female subjects. Cancer Epidemiol. Biomarkers Prev. 6, 597-601.
48. Chen, H.-J.; Hong, C.-L.; Wu, C.-F.; Chiu, W.-L. (2003) Effect of smoking on urinary 3,N4-ethenocytosine levels measured by gas chromatography/mass spectrometery. Toxicol. Sci. 76, 321-327.
49. Basu, A. K.; Wood, M. L.; Niedernhofer, L. J.; Ramos, L. A.; Essigmann, J. M. (1993) Mutagenic and genotoxic effect of three vinyl chloride-induced DNA lesions : 1N6ethenoadenine, 3,N4-ethenocytosine, and 4-amino-5-(imidazol-2-yl)imidazole. Biochemistry 32, 12793- 12801.
50. Moriya, M.; Zhang, W.; Johnson, F.; Grollman, A. P. (1994) Mutagenic potency of exocyclic DNA adducts: marked differences between
Escherichia coli and simian kidney cells. Proc. Natl. Acad. Sci. U.S.A. 91, 11899-11903.
51. Pandya, G. A.; Moriya, M. (1996) 1,N6-ethenodeoxyadenosine,a DNA adduct highly mutagenic in mammalian cells. Biochemistry 35, 11487-11492.
52. Saparbaev, M.; Langoue, S.; Privezentzev, C.; Guengerich, P. F.; Cai, H,; Elder R,; and Laval, J, (2002) 1,N2-ethenoguanine, a mtagenic DNA adduct, Is a primary substrate of escherichia coli mismatch-specific uracil-DNA glycosylase and human alkylpurine-DNA-N-glycosylase J. Biol. Chem. 277, 26987–26993.
53. Bartsch, H.; Nair, J.; Velic, I. (1997) Etheno-DNA base adducts as tools in human cancer aetiology and chemoprevention. Eur. J. Cancer Prev. 6, 529-534.
54. Ames, B. N.; Gold, L. S.; Willett, W. C. (1995) The causes and prevention of cancer. Proc. Natl. Acad. Sci. U.S.A. 92, 5258-5265.
55.Gackowski, D., Speina, E., Zielinska, M., Kowalewski, J., Rozalski, R., Siomek, ., Paciorek, T., Tudek, B., and Olinski, R. (2003) Products of oxidative DNA damage and repair as possible biomarkers of susceptibility to lung cancer. Cancer Res. 63, 4899-4902.
56.Speina, E., Zielinska, M., Barbin, A., Gackowski, D., Kowlewski, J., Graziewicz, M. A., Siedlecki, J. A., Olinski, R., and Tudel, B. (2003) Decreased repair activities of 1,N6-ethenoadenine and 3,N4-ethenocytosine in lung adenocarcinoma patients. Cancer Res. 63, 4351-4357.
57. Matijasevie, Z.; Sekiguchi, M.; Ludlum, D. B. (1992) Release of N2,3-ethenoguanine from chloroacetaldehyde-treated DNA by Escherichia coli 3-methyladenine DNA glycosylase II. Proc. Natl. Acad. Sci., 89, 9331-9334.
58. Hang, B.; Chenna, A.; Rao, S.; Singer, B. (1996) 1,N6-Ethenoadenine and 3,N4-ethenocytosine are excised by separate human DNA glycosylases. Carcinogenesis 17, 155-157.
59. Saparbaev, M.; Laval, J. (1998) 3,N4-Ethenocytosine, a highly mutagenic adduct, is a primary substrate for Escherichia coli double-stranded uracil-DNA glycosylase and human mismatch-specific thymine-DNA glycosylase. Proc. Natl. Acad. Sci. U.S.A. 95, 8508-8513.
60. Hang, B.; Medina, M.; Fraenkel-Conrat, H.; Singer, B. (1998) A 55-kDa protein isolated from human cells shows DNA glycosylase activity toward 3,N4-ethenocytosine and the G/T mismatch. Proc. Natl. Acad. Sci. U.S.A. 95, 13561-13566.
61.Fortini, P., Parlanti, E., Sidorkina, O.M., Laval, J., Dogliotti, E., (1999) The type of DNA glycosylase determines the base excision repair in mammalian cells. J. Biol. Chem. 274, 15230–15236.
62.Kochetkov, N.K., Shibaev, V.N., Kost, A.A., (1971) New reaction of adenine and cytosine derivatives, potentially useful for nucleic acid modifications. Tetrahedron Lett. 22, 1993–1996.
63.Bartsch, H., Barbin, A., Marion, M.-J., Nair, J., Guichard, Y., (1994) Formation, detection, and role in carcinogenesis of ethenobases in DNA. Drug Metab. Rev. 26, 349–371.
64.Ranasinghe, A., Scheller, N., Wu, K.Y., Upton, P.B., Swenberg J.A., (1998) Application of gas chromatography/electron capture negative chemical ionization on high-resolution mass spectrometry for nalysis of DNA and protein adducts. Chem. Res. Toxicol. 11 (5). 520–526.
65.Swenberg, J.A., Bogdanffy, M., Ham, A., Holt, S., Kim, A., Morinello, E.J., Ranasinghe, A., Scheller, N., Upton, P.B., (1999) Formation and repair of DNA adducts in vinyl chloride- and vinyl fluoride-induced carcinogenesis. In: Singer, B., Bartsch, H. Eds., Exocyclic DNA Adducts in Mutagenesis and Carcinogenesis, vol. 150. IARC Science Publisher, IARC, Lyon, pp. 29–43.
66.Holt, S., Yen, T.Y., Sangaiah, R., Swenberg, J.A., (1998) Detection of 1,N6-ethenoadenine in rat urine after chloroethylene exposure. Carcinogenesis 19, 1763–1769.
67.Nair, J., Barbin, A., Guichard, Y., Bartsch, H., (1995) 1,N6- Ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine in liver DNA from humans and untreated rodents detected by immunoaffinity/32P-postlabeling. Carcinogenesis 16, 613– 617.
68.Nair, J., Barbin, A., Velic, I., Bartsch, H., (1999) Etheno DNA-base adducts from endogenous reactive species. Mutat. Res. 424, 59–69.
69.Ham, A.J., Ranasinghe, A., Koc, H., Swenberg, J.A., (2000) 4-hydroxy-2-nonenal and ethyl linoleate form N2,3- ethenoguanine under peroxidizing conditions. Chem. Res. Toxicol. 13 (12), 1243–1250.
70.Singer, B., Spengler, S.J., Chavez, F., Kusmierek, J.T., (1987) The vinyl chloride-derived nucleoside, N2,3-ethenoguanosine, is a highly efficient mutagen in transcription. Carcinogenesis 8, 745–747.
71. Gonzalez-Reche, L.M., Koch H.M., Weiß T., Mu¨ ller J., Drexler H., Angerer J.,* (2002) Analysis of ethenoguanine adducts in human urine using high performance liquid chromatography–tandem mass spectrometry. Toxicology Letters 134, 71–77.
72. Chen, H. J. C., Chiu, W. L., (2003) Detection and Quantification of 1,N6-Ethenoadenine in Human Urine by Stable Isotope Dilution Capillary Gas Chromatography/Negative Ion Chemical Ionization/Mass Spectrometry. Chem. Res. Toxicol. 16, 1099-1106
73.Lindahl, T., Nyberg, B., (1972) Rate of depurination of native deoxyribonucleic acid. Biochemistry. 11, 3610-3618.
74.Singer, B., Grunberger, D., (1983) Depurination and depyrimidination. Molecular Biology of Mutagens and Carcinogens, pp 16-19, Plenum Press, New York.
75.Boiteuz, S., Laval, J., (1982) Coding properties of poly-(deoxycytidylic acid) templates containing uracil or apyrimidinic sites: in vitro modulation of mutagenesis by deoxyribonucleic acid repair enzymes. Biochemistry. 21, 6746-6751.
76.Loeb, L., Preston, B. D., (1986) Mutagenesis by apurinic/apyrimidinic sites. Ann. Rev. Genetics. 20, 201-230.
77. Wilde, J. A., and Bolton, P. H. (1989) Characterization of the equilibrating forms of the aldehydic abasic site in duplex DNA by 17O NMR. J. Am. Chem. Soc. 111, 1894-1896.
78. Talpaert-Borle´, M., and Liuzzi, M. (1983) Reaction of apurinic/ apyrimidinic sites with [14C]methoxyamine, a method for the quantitative assay of AP sites in DNA. Biochim. Biophys. Acta 740, 410-416.
79.Talpaert-Borle´, M., and Liuzzi, M. (1984) A process for directly determining apurinic and apyrimidinic sites in DNA. Eur. Pat. Appl. 84103189.1.
80.Livingston, D. C. (1964) Degradation of apurinic acid by condensation with aldehyde reagents. Biochim. Biophys. Acta 87, 538-540.
81.Coombs, M. M., and Livingston, D. C. (1969) Reaction of apurinic acid with aldehyde reagents. Biochim. Biophys. Acta 174, 161- 173.
82. Kubo, K., Ide, H., Wallace, S. S., and Kow, Y. W. (1992) A novel sensitive and specific assay for abasic sites, the most commonly produced DNA lesion. Biochemistry 31, 3703-3708.
83. Chen, B. X., Kubo, K., Ide, H., Erlanger, B. F., Wallace, S. S., and Kow, Y. W. (1992) Properties of a monoclonal antibody for the detection of abasic sites, a common DNA lesion. Mutat. Res. 273, 253-261.
84.Kubo, K., Ide, H., Wallace, S. S., and Kow, Y. W. (1992) A novel sensitive and specific assay for abasic sites, the most commonly produced DNA lesion. Biochemistry 31, 3703-3708.
85.Ide, H., Akamatsu, K., Kimura, Y., Michiue, K., Makino, K., Asaeda, A., Takamori, Y., and Kubo, K. (1993) Synthesis and damage specificity of a novel probe for the detection of abasic sites in DNA. Biochemistry 32, 8276-8283.
86.Shapiro, R., Pohl, S. H. (1968) The reaction of riosenucleosides with nitrous acid. Side products and kinetics. Biochem. 7, 448-455.
87.Burney, S., Caulfield, J. L., Niles, J. C., Wishnok, J. S., Tannenbaum, S. R. (1999) The chemistry of DNA damage from nitric oxide and peroxynitrite. Mutat. Res. 424, 37-49.
88.Morinello, E. J., Ham, A-J. L., Ranasinghe, A., Sangaiah, R., and Swenberg, J. A. (2001) Simultaneous Quantitation of N2,3-Ethenoguanine and 1,N2-ethenoguanine with an immunoaffinity/gas chromatography/high-resolution Mass spectrometry assay. Chem. Res. Toxicol. 14, 327-33
89.Chen, H.-J. C., Chiang, L. C., Txeng, M. C., Zhang, L. L., Ni, J., and Chung, F. L., (1999) Detection and quantification of 1,N6-ethenoadenine in human placental DNA by mass spectrometry. Chem. Res. Toxicol. 12, 1119-1126.
90.Feynman, R. P., (1991) Science, 254, 1300
91.Brus, L. E., Efros, A. L., Itoh, T., (1996) J. Lumin. 70, R7-R8.
92.Weller, H., (1996) Philo. Trans. R. Soc. London, Ser. A. 354, 757.
93.Kalyanasundaram, K., Borgarello, E., Duonghong, D., Gratzzel, M., (1981) Angew. Chem. Int. Ed. Engl. 20, 933.
94.Stpanyuk, V. A., Echer, Hergert, W., Rennert, P., Wilderger, K., Zeller, R., Dederichs, P. H., (1997) Surf. Sci. 377, 495.
95.Buffat, Ph., Borel, J. P., (1976) Phys. Rev. A 13, 2287.
96.Golden, A. N., Echer, C. M., Alivisatos, A. P., (1992) Science. 236, 1425.
97.Creighton, J. A., Eadon, D. G. J., (1991) Chem. Soc. Faraday. Trans. 87, 3881
98.張仕欣、王崇人, (1998) 化學, 56, 209.
99.Bruckner, C. A., Synovec, R. E., (1996) Talanta, 43, 901-907.
100.Okada, R., (1991) Phys.Chem. 58, 1662.
101.Henglein, A., (1993) J. Phys. Chem. 97, 5445.
102.Wang, D. S., Ketxer, M., Lee, C. L., Wang C. R. C., (1981) Phys. Rev. B. 24, 1777.
103.Boturyn, D., Boudali, A., Constant, J.-F., Dcfrancq, E., and Lhomme, J., (1997) Sythesis of Fluorescent Probes for the Detection of Abasic Sites in DNA. Tetrahedron. 53, 5485-5492.
104.Morrow, J. D.; Frei, B.; Longmire, A. W.; Gaziano, J. M.; Lynch, S. M.; Shyr, Y.; Strauss, W. E.; Oates, J. A.; Roberts, L. J. (1995) Increase in circulating products of lipid peroxidation (F2-isoprostanes) in smokers. Smoking as a cause of oxidative damage. N. Engl. J. Med. 332, 1198-1203.