Journal of Environmental Health Sciences (한국환경보건학회지)

Korean Society of Environmental Health (한국환경보건학회)
권호 표지

A Study on 10 Metabolites Separated from DNA Adduce of Blood Lymphocytes in Rats Exposed Orally with 3,3-dichlorobenzidine(DCB) by GC/MS-SIM

  • Published : 2002.11.01
Download PDF
⟨ Previous Next ⟩

Abstract

3.3'-Dichlorobenzidine(DCB) has be shown carcinogenic in several animals, and the development of non-invasive biomonitoring method in workers exposed with it is a very important subject. DNA adduct is a good biomarker for biomonitoring about carcinogens exposure, and lymphocytes is a good non-invasive samples. So we studied to analyze metabolites in blood lymphocytes of female Sprague-Dawley rats exposed orally with DCB(20, 30, and 40 mg/kg wt.) for 3 weeks. For analysis of them, we isolated DNA adducts from blood lymphocytes by using the enzymes method in /sup 32/P-postlabeling, and measured them by using gas chromatography/mass spectrometry-selected ion monitoring(GC/MS-SIM). 4-aminobiphenyl and phenanthrene-d/sub 10/ were added as internal standard for blank sample. Standard metabolites of DCB were synthesized with using pyridine and acetic acid which were promoter and controller in acetylation of DCB. And they were used for calibration curve. Our results showed two kinds of metabolites in DNA adducts of blood lymphocytes. They were N-acetyl 3,3'-dichlorobenzidine(acDCB) and N,N'-diacetyl 3,3'-dichiorobenzidine(di-acDCB ). They were combined with DNA at the same time as an acetyl of it was removed. So we measured DCB and acDCB for two kinds of metabolites in DNA adducts of blood lymphocytes. Our results showed the levels of DCB were 1.46∼2.26 times more than that of acDCB. And also the levels of metabolites in 20, 30 and 40 mg/kg wt. were gradually increased with going days from 1st to 3rd week. They are 1.66, 1.38 and 0.90 times in total metabolites, 1.76, 1.49 and 1.02 times in DCB, and 1.51, 1.22 and 1.28 times in acDCB. In conclusion, the results of this study showed DCB exposed to rats formed DNA adduct in blood lymphocytes after acetylated to N-acetyl 3.3'-dichloro benzidine(acDCB) and N,N'-diacetyl 3,3'-dichlorobenzidine(di-acDCB), and they could be analyzed by us ing gas chromatography/mass spectrometry-selected ion monitoring(GC/MS-SIM).

Keywords

References

  1. Albrecht, W. and Neumann, H.G. : Biomonitoring of aniline and nitrobenzene. Arch. Toxicol., 57, 1-5, 1985 https://doi.org/10.1007/BF00286566
  2. Birner, G., Albrecht, W. and Neumann, H.G.: Biomonitoring of aromatic amines: iii hemoglobin binding of benzidine and some benzidine conge-ners. Arch. Toxicol., 64(2), 97-102, 1990 https://doi.org/10.1007/BF01974393
  3. DHHS : Toxicological profile for 3,3'-dichloroben-zidine, US Department of Health & Human Ser-vices, Public Health Services, Agency for Toxic Substances and Disease Registry, Draft for Public Comment, p.1-123, Feb. 1998
  4. IARC : Monographs of carcinogenic risk of chem-icals to human: some industrial chemicals and dye-stuffs. 3,3'-dichlorobenzidine and its hydrochloride. Int. Agency Res. Cancer, 1982
  5. Eyer, G. and Gallemann, D. and Patai, S.: The chemistry of amino, nitroso, nitro and related groups, Wiley, New York, p.999, 1996
  6. Joppich-Kuhn, R., Hanggi, R., Sagelsdorff, P., Smith, A.E., Weide1i, H.J. and Joppich, M.: Deter-mination of dichlorobenzidine-hemoglobin adducts by GC/MS-NCI. Int. Arch. Occup. Environ. Health, 69, 240-246, 1997 https://doi.org/10.1007/s004200050142
  7. Lee, I.M. : Human physiology. Yongsul ed. 128-155, 1992
  8. Lee, J.H., Shin, H.S. and Jung, D.G. : Synthesis and analysis of N-acetyl-DCB and N,N'-diacetyl-DCB, which were urinary metabolites of 3,3'-dichloroben-zidine(DCB). Not Yet Published, 2002
  9. Neumann, H.G. in Berlin A, Draper M., Hem-minki, K., Vainio, H.(Eds) : Monitoring human exposure to carcinogenic and mutagenic agents (IARC Scientific Publications No.59). International Agency for Research on Cancer, Lyon, p.115(1984)
  10. Pliss, G.B.: Dichlorobenzidine as blastomogenic agent. Vapr Oncol., 5, 524-533, 1959
  11. Reh, B.D., DeBord, D.G., Butler, M.A., Reid, T.M., Mueller, C. and Fajen, J.M.: O super(6)-meth-ylguanine DNA adducts associated with occupa-tional nitrosamine exposure. Carcinogenesis, 21(1), 29-33, 2000 https://doi.org/10.1093/carcin/21.1.29
  12. Reid, T.M., Morton, K.C. and Wang, C.Y. : Mutage-nicity of some benzidine congers and their N-acety-lated and N,N-diacetylated derivatives in different strains of Salmonella typhimurium. Environ. Mutagen., 6, 145-151, 1984 https://doi.org/10.1002/em.2860060205
  13. Sabbioni, G. and Beyerbach, A. : 'Hemoglobin adducts of aromatic amines: diamines and polyaromatic amines. J. of Chromatography B, 744, 377-387, 2000 https://doi.org/10.1016/S0378-4347(00)00265-6
  14. Sellakumar, A.R., Montesano, R. and Saffiotti, U. : Aromatic amines carcinogenicity in hamsters. Proc. Am. Assoc. Cancer Res., 10, 78, 1969
  15. Stula, E.F., Sherman, H., Zapp, J.A. and Clayton J.W. : Experimental neoplasia in rats from oral administration of 3,3'-dichlorobenzidine, 4,4'-meth-ylene-bis(2-chloroani line) and. Toxicol. Appl. Phannacol., 31, 159-176, 1975 https://doi.org/10.1016/0014-2999(75)90090-4
  16. Stula, E.F., Barnes, J.R., Sherman, H., Reinhardt, C.F., Zapp, J.A, : Liver and urinary bladder tumors in dogs from 3,3'-dichlorobenzidine. J. Environ. Pathol. Taxicol., 1, 475-490, 1978
  17. Talaska, G., Underwood, P., Maier, A., Lewtas, J., Rothman, N. and Jaeger, M. : Polycyclic aromatic hydrocarbons(PAHs), nitro-PAHs and related envi-ronmental compounds: biological markers of expo-sure and effects. Environmental Health Perspectives, 104(5), 901-906, 1995 https://doi.org/10.2307/3433008
  18. Tanaka, K. : Urinary metabolites of 3,3'-dichloro-benzidine and their mutagenicity. Sangyo Igaku (Japanese)', 23, 426-427, 1981 https://doi.org/10.1539/joh1959.23.426
  19. Zharkov, D.O., Gilboa, R., Yagil, I., Kycia, J.H., Gerchman, S.E., Shoham, G. and Grollman, A.P. : Role for lysine 142 in the excision of adenine from A:G mispairs by MutY DNA glycosy1ase of Escherichia coli. Biochemistry, 39, 14768-14778, 2000 https://doi.org/10.1021/bi001538k