Effects of pH and Structure on the Mutagenic Activity of N-Nitroso Compounds

  • Joseph B. Guttenplan
Part of the Topics in Chemical Mutagenesis book series (TCM, volume 1)


N-nitroso compounds comprise a class of chemicals containing a wide variety of carcinogens and mutagens.(1–4) These compounds have been of theoretical and practical interest to researchers for a number of reasons, such as their organo-tropic tumor induction, the large variability in the potencies and target organs of various N-nitroso compounds, the relatively simple metabolism and breakdown of the lower molecular weight compounds, and the fact that some members of this class of compounds are environmental chemicals or can be formed in vivo from nitrite and secondary or tertiary amines under acidic conditions.(1–4)The contribution to human cancer caused by continued exposure to low levels of N-nitroso compounds is uncertain, but exposure of pregnant mice to levels of 10 ppb of DMN resulted in significant lung tumor formation in the male progeny.(5) A generalized scheme for the metabolism and decomposition of N-nitroso compounds is given in Figure 1.(2)


Mutagenic Activity Chemical Carcinogen Nitroso Compound Urea Urethane Mutagenesis Assay 
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Abbreviations used in this chapter




colony-forming units




















mixed function oxidase(s)
















9000 × g supernatant


sodium dodecylsulfate


0.15 M NaCI + 0.15 M Na3 citrate. pH 7.0.


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  1. 1.
    P. N. Magee, and J. M Barnes, Carcinogenic nitroso compounds, Adv. Cancer Res. 10, 163–256 (1967).PubMedCrossRefGoogle Scholar
  2. 2.
    H. Druckrey, R. Preussmann, S. Ivankovic, and D. Schmäht, Organtrope carcinogene wirkungen bei 65 verschiedene N-nitroso-verbindungen an BD-Ratten, Z. Krebsforsch. 64, 103–201 (1967).Google Scholar
  3. 3.
    P. N. Magee, R. Montesano, and R. Preussmann, N-Nitroso compounds and related carcinogens, in: Chemical Carcinogens (C. Searle, ed.), ACS Monograph 173, pp. 491–625, American Chemical Society, Washington, D.C. (1976).Google Scholar
  4. 4.
    E. J. Olajos and F. Coulston, Comparative toxicology of N-nitroso compounds and their carcinogenic potential to man, Ecotoxic. Enriron. Safety 2, 317–367 (1978).CrossRefGoogle Scholar
  5. 5.
    L. M. Anderson, L. J. Priest, and J. M. Budinger, Lung tumorigenesis in mice after chronic exposure in early life to a low dose of dimethylnitrosamine, J. Natl. Cancer Inst. 62, 1553–1555 (1979).PubMedGoogle Scholar
  6. 6.
    P. D. Lawley, Carcinogenesis by alkylatingagents. in: Chemical Carcinogens(C. E. Searle, ed.), ACS Monograph 173. pp. 83–244, American Chemical Society, Washington, D. C. (1976).Google Scholar
  7. 7.
    B. Singer, N-Nitroso alkylating agents; formation and persistence of alkyl derivatives in mammalian nucleic acids as contributing factors in carcinogenesis, J. Natl. Cancer Inst. 62, 1329–1339 (1979).PubMedGoogle Scholar
  8. 8.
    J. H. Weisburger, Bioassays and tests for chemical carcinogens, in: Chemical Carcinogens (C. E. Searle, ed.). ACS Monograph 173, pp. 1–23, American Chemical Society, Washington, D. C. (1976).Google Scholar
  9. 9.
    E. C. Miller and J. A. Miller, The metabolism of chemical carcinogens to reactive electrophiles and their possible mechanisms of action in carcinogenesis, in: Chemical Carcinogens (C. E. Searle, ed.), ACS Monograph 173, pp. 736–762, American Chemical Society, Washington, D. C. (1976).Google Scholar
  10. 10.
    L. L. Gerchman and D. B. Ludlam, The properties of 06-methylguanine in templates for RNA polymerase, Biochim. Biophrs. Acta 308, 310–316 (1973).Google Scholar
  11. 11.
    P. J. Abbott and R. Saffhill, DNA synthesis with methylated poly (dC-dG) templates: Evidence for a competitive nature to miscoding by O6methylguanine, Biochim. Biophrs. Acta. 562, 51–61 (1979).Google Scholar
  12. 12.
    J. W. Nicoll, P. F. Swann, and A. E. Pegg, Effects of dimethylnitrosamine on persistence of methylated guanines in rat liver and kidney DNA, Nature (London) 254, 261–262 (1975).CrossRefGoogle Scholar
  13. 13.
    R. Montesano, H. Bresil, P. M. Ghyslaine, G. P. Margison, and A. E. Pegg, Effect of chronic treatment of rats with dimethylnitrosamine on the removal of O6-methylguanine from DNA, Cancer Res. 40, 452–458 (1980).PubMedGoogle Scholar
  14. 14.
    J. Buechler and P. Kleihues, Excision of O6-methylguanine from DNA of various mouse tissues following a single injection of N-methyl-N-nitrosourea, Chem.-Biol. Interactions 16, 325–333 (1977).CrossRefGoogle Scholar
  15. 15.
    P. Kleihues, S. Bamborschke, and G. Doerjer, Persistence of alkylated DNA bases in Mongolian gerbil (Meriones unguiculatus)following a single dose of methylnitrosourea, Carcinogenesis 1, 111–113 (1980).PubMedCrossRefGoogle Scholar
  16. 16.
    B. L. Van Duuren, Tumor-promoting and co-carcinogenic agents, in: Chemical Carcinogens (C. E. Searle, ed.), ACS M onograph 173, pp. 24–51, American Chemical Society, Washington. D. C. (1976).Google Scholar
  17. 17.
    H. C. Pitot, Biological and enzymatic events in chemical carcinogenesis, Ann. Rev. Med. 30, 25–29 (1979).PubMedCrossRefGoogle Scholar
  18. 18.
    D. Jenssen, B. Beije, and C. Ramel, Mutagenicity testing on Chinese Hamster V79 cells treated in the in vitro liver perfusion system. Comparative investigation of different in vitro metabolizing systems with dimethylnitrosamine and benzo(a)pyrene, Chem.-Biol. Interactions 27, 27–39 (1979).CrossRefGoogle Scholar
  19. 19.
    K. S. Bakshi and D..I. Brusick, Bioactivation of DMN in intrasanguinous host-mediated assay in male and female mice, Environ. Mutagenesis 1, 150 (1979).Google Scholar
  20. 20.
    J. McCann, E. Choi, E. Yamasaki, and B. N. Ames, Detection of carcinogens as mutagens in the Salmonella, microsome test: Assay of 300 chemicals. Proc. Natl. Acad. Sci. USA 72,5135–5139(1975).PubMedCrossRefGoogle Scholar
  21. 21.
    B. N. Ames, J. McCann, and E. Yamasaki, Methods for detecting carcinogens and mutagens with the Salmonella,mammalian microsome mutagenicity test, Mutat. Res. 31.347–364(1975).PubMedGoogle Scholar
  22. 22.
    J. McCann and B. N. Ames, Detection of carcinogens as mutagens in the Salmonella micro-some test: Assay of 300 chemicals: Discussion, Proc. Natl. Acad. Sci. USA 73, 950–954 (1976).PubMedCrossRefGoogle Scholar
  23. 23.
    E. C. Miller and J. A. Miller, The mutagenicity of chemical carcinogens, correlations, problems and interpretations, in: Chemical Mutagens, Principles and Methods for Their Detection (A. Hollaender, ed.), Vol. 1, pp. 83–119, Plenum, New York (1971).Google Scholar
  24. 24.
    H. V. Mailing, Dimethylnitrosamine: Formation of mutagenic compounds by interaction with mouse liver microsomes, Mutat. Res. 13, 425–429 (1971).CrossRefGoogle Scholar
  25. 25.
    P. Czygan, H. Greim, A. J. Garro, F. Hutterer, F. Schaffner, H. Popper, O. Rosenthal, and D. Y. Cooper, Microsomal metabolism of dimethylnitrosamine and the cytochrome P-450 dependency of its activation to a mutagen, Cancer Res 33, 2983–2986 (1973).PubMedGoogle Scholar
  26. 26.
    J. C. Arcos, D. L. Davies, C. F. L. Brown, and M. F. Argus, Repressible and inducible enzymic forms of dimethylnitrosamine-demethylase. Z. Krehs%orsch 86, 171–183 (1976).Google Scholar
  27. 27.
    L G. Sipes, M. L. Slocumb, and G. Holtzman, Stimulation of microsomal dimethylnitrosamine-N-demethylase by pretreatment of mice with acetone, Chem. Biol. Interactions 21. 155–166(1978).Google Scholar
  28. 28.
    A. Wood, W. Levin, A. Y. Lu, H. Yagi, O. Hernandez, D. M. Jerina, and A. H. Conney. Metabolism of benzo(a)pyrene and benzo(a)pyrene derivatives to mutagenic products by highly purified hepatic microsomal enzymes, J. Biol. Chen. 251, 4882–4890 (1976).Google Scholar
  29. 29.
    S. Milstein, and J. B. Guttenplan, unpublished results.Google Scholar
  30. 30.
    J. B. Guttenplan, Comutagenic effects exerted by N-nitroso compounds, Mutai. Res. 66, 25–32 (1979).CrossRefGoogle Scholar
  31. 31.
    J. B. Guttenplan, Detection of trace amounts of dimethylnitrosamine with a modified Salmonella’ microsome mutagenicity assay, Mutai. Res. 64, 91–94 (1979).Google Scholar
  32. 32.
    J. B. Guttenplan, Enhanced mutagenic activities of N-nitroso compounds in weakly acidic media, Carcinogenesis 1, 439–444 (1980).PubMedCrossRefGoogle Scholar
  33. 33.
    T. Yahagi, M. Nagao, Y. Seino, T. Matsushima. T. Sugimura. and M. Okada. Mutagenicities of N-nitrosamines on Salmonella, Mutai. Res. 48, 121–130 (1977).CrossRefGoogle Scholar
  34. 34.
    S. Milstein and J. B. Guttenplan, Near quantitative production of molecular nitrogen from metabolism of dimethylnitrosamine, Biochem. Biophrs. Res. Commun. 87, 337–342 (1979).CrossRefGoogle Scholar
  35. 35.
    J. L. Frei, D. H. Swenson, W. Warren, and P. D. Lawley, Alkylation of deoxyribonucleic acid in vira in various organs of C57BL mice by the carcinogens N-mcthyt-N-nitrosourea. N-ethylN-nitrosourea and ethylmethanesulfonate in relation to induction of thymic lymphoma, Biochem. J. 174, 1031–1044 (1978).PubMedGoogle Scholar
  36. 36.
    E. R. Garrett, S. Goto, and J. B. Stubbins, Kinetics of solvolyses of various N-alkyl-Nnitrosoureas in neutral and alkaline solutions, J. Pharm. Sci. 54, 119–l23 (1965).PubMedCrossRefGoogle Scholar
  37. 37.
    M. Mochizuki, T. Ant o, T. Sone, and M. Okada. Isolation and characterization of alphahydroxydialkylnitrosamines, in: Proceedings of the 38th Annual Meeting of the Japanese Cancer Association. p. 321. Japanese Cancer Association, Tokyo (1979).Google Scholar
  38. 38.
    f. Wang, T. Kakizoe, P. Oion, T. Furrer, A. J. Varghese and W. R. Bruce. Volatile nitrosamines in normal human faeces, Nature 276, 280–281 (1978).PubMedCrossRefGoogle Scholar
  39. 39.
    J. B. Guttenplan, R. Bonomo, and.J. Fliessbach, Recovery of bento(a)pyrcne(BP) metabolites and dimethylnitrosamine in rat saliva. Proc. Am. Assoc. Cancer Res. 21. 124 (1980).Google Scholar
  40. 40.
    J. Fliessbach and J. B. Guttenplan, unpublished results.Google Scholar
  41. 41.
    M. J. Privai, V. D. King. and A. T. Sheldon, Jr., The mutagenicity of dialkylnitrosamines in the Salmonella plate assay, Environ. Mutagenesis 1. 95–104 (1979).CrossRefGoogle Scholar
  42. 42.
    H. Bartsch. A. Camus, and C. Malaveille, Comparative mutagenicity of N-nitrosamines in a semi-solid and in a liquid incubation system in the presence of rat or human tissue fractions. Mutai. Res. 37. 149–162 (1976).CrossRefGoogle Scholar
  43. 43.
    J. Unger and.J. B. Guttenplan. Kinetics of benzo(a)pyrene induced mutagenesis in a highly sensitive Salmonella; microsome assay, Mutai. Res. 77, 221–228 (1980).CrossRefGoogle Scholar
  44. 44.
    J. McCann, N. F. Spingarn, J. Kobari, and B. Ames, The detection of carcinogens as mutagens: Bacterial tester strains with R factor plasmids, Proc. Natl. Acad. Sci. (USA)271, 5–13 (1975).Google Scholar
  45. 45.
    W. Warren and P. D. Lawley, The removal of alkylation products from DNA of Esc/terichia coli cells treated with the carcinogens N-ethyl-N-nitrosourea and N-methyl-N-nitrosourea: Influence of growth conditions and DNA repair defects, Carcinogenesis 1. 67–78 (1980).PubMedCrossRefGoogle Scholar
  46. 46.
    G. P. Margison, J. A. Swindwell, L. H. Ockey. and A. W. Craig, The effects of a single dose of dimethylnitrosamine in the Chinese hamster and the persistence of DNA alkylation products in selected tissues, Carcinogenesis 1. 91–96 (1980).PubMedCrossRefGoogle Scholar
  47. 47.
    D. E. Ryan, P. F. Thomas, D. Kornzeniowski, and W. Levin, Separation and characterization of highly purified forms of liver microsomal cytochrome P-450 from rats treated with polychlorinated biphenyls, phenobarbitol and 3-methylcholanthrene, J. Biol. Chem. 254. 1365–1374 (1979).PubMedGoogle Scholar
  48. 48.
    J. B. Farrelly, A new assay for the microsomal metabolism of nitrosamines, Cancer Res. 40, 3241–3244 (1980).PubMedGoogle Scholar
  49. 49.
    T. K. Rao, A. A. Hardigree, J. A. Young, W. Lijinsky, and J. L. Epler. Mutagenicity of N-nitrosopiperidines with Salmonella irphimuriuml microsomal activation system, Mutat. Res. 56, 131–145 (1977).CrossRefGoogle Scholar
  50. 50.
    W. Lijinsky, Structure-activity relations in carcinogenesis by N-nitroso compounds. Chapter 10, this book.Google Scholar
  51. 51.
    T. K. Rao, J. A. Young. W. Lijinsky, and J. L. Epler. Mutagenicity of aliphatic nitrosamines in Salmonella ttphimurium, Mural. Res. 66,1–7 (1979).Google Scholar
  52. 52.
    W. Lijinsky, et al.,personal communication.Google Scholar

Copyright information

© Plenum Press, New York 1984

Authors and Affiliations

  • Joseph B. Guttenplan
    • 1
  1. 1.Department of BiochemistryNew York University Dental CenterNew YorkUSA

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