Non-invasive Biological Monitoring of DNA Adducts Formed at Workers Handling 3,3-Dichlorobenzidine(DCB) by Using GC/MS

  • Lee, Jin-Heon (Dept. of Environmental Education, Kongju National University)
  • Published : 2003.12.01


We examine the metabolites(DCB and acetyl DCB) extracted from exfoliated urothelial cells of 33 workers who employed DCB-handling industries. The characteristics of workers submitted urine, whose age, working years and smoking persons were 41.9$\pm$11.1, 8.7$\pm$5.5 and 25(32.0%), respectively. DNA adduct was isolated from the exfoliated urothelial cells by applying $^{32}$ p-postlabeling procedure. Metabolites(DCB and acetyl DCB) were extracted from DNA adducts by hydrolyzing and N-glycosylase. Concentrations of DCB and acetyl DCB were 28.6$\pm$5.25 ng/g DNA, and 17.0$\pm$3.73 ng/g DNA, respectively. The regression between DCB level and exposure years of workers is y = 1.668 + 2.588x(p = 0.005, $r^2$= 0.394). The regression between acetyl DCB level and exposure years of workers is y = 8.071 + 1.325x(p = 0.076, $r^2$= 0.222). Smoking workers are significantly higher than non-smoking workers on DCB and acetyl DCB level(p = 0.065 and 0.021, respectively). DCB level was 33.9$\pm$7.14 ng/g DNA on smokers, and 23.1$\pm$9.97 ng/g DNA on non-smokers. Acetyl DCB was 25.1$\pm$5.27 ng/g DNA on smokers, and 8.92$\pm$7.22 ng/g DNA on non-smokers.


3,3-dichlorobenzidine(DCB);acetyl DCB;GC/MS;DNA adduct Exfoliated urothelial cells


  1. Lee, J.H. and Shin, H.S. : Determination of hemoglobin adducts formed in rats exposed orally with 3,3'-dichlorobenzidine by GS/MS-SIM, Toxicology and Industrial Health, 18, 191-199, 2002
  2. Lee, J.H., Shin, H.S., Jung, D.G. and Lee, Y.E.: Urinary monitoring method of 3,3'-dichlorobenzidine (DCB) and its metabolites, N-acetyl DCB and N,N'-diacetyl DCB, Industrial Health(Japane), 41, 242-248, 2003
  3. Talaska, G., Dooley, K.L. and Kadlubar, F.F.: Detection and characterization of carcinogen-DNA adducts in exfoliated urothelial cells from 4-aminobiphenyl-treated dogs by 32P-postlabeling and subsequent thin-layer and high-pressure liquid chromatography, Carcinogenesis, 11(4), 639-646, 1990
  4. Tanaka K.: Urinary metabolites of 3,3'-dichlorobenzidine and their mutagenicity, Sangyo Igaku (Japanese), 23, 426-427, 1981
  5. DHHS: Toxicological profile for 3,3'-dichlorobenzidine, US Department of Health & Hurnan Services, Public Health Services, Agency for Toxic Substances and Disease Registry, Draft for Public Comment, 1-123, 1998
  6. Matanoski, G.M. and Elliot, E.A. : Bladder cancer epidemiology, Epidemiol. Rev. 3, 203-229, 1981
  7. Birner, G., Albrecht, W. and Neumann, H.G.: Biomonitoring of aromatic amines: iii hemoglobin binding of benzidine and some benzidine congeners, Arch Toxickol, 64(2), 97-102, 1990
  8. Gupta, R.C. : Non-random binding of the carcinogen N-hydroxy-2-acetyl aminofluorene to repetitive sequences of rat liver DNA in vivo, Proc. Natl. Acad. Sci. USA, 81, 6943-6947, 1984
  9. Bi, W., Hayes, R.B., Feng, P., Qi, Y., You, X., Zhen, J., Zhang, M., Qu., B.Q., Fu., Z., Chen, M., Chien, H.T.C and Blot, W. : Mortality and incidence of bladder cancer in benzidine-exposed workers in China, American Journal of Industrial Medicine, 21, 481-489, 1992
  10. 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'-methylene-bis(2-chloroani line). Toxicol Appl Pharmacol, 31, 159-176, 1975
  11. Pliss, G.B.: Dichlorobenzidine as blastomogenic agent. Vapr Oncol, 5, 524-533, 1959
  12. IARC: Monographs of carcinogenic risk of chemicals to human: some industrial chemicals and dyestuffs 3,3'-dichlorobenzidine and its hydrochloride. Int Agency Res Cancer, 100, 1982
  13. Synthesis and isolation of monoacetyl DCB and diacetyl DCB from 3,3'-dichlorobenzidine, Kor. J Env. Hlth., 29(2), 50-55, 2003
  14. Sellakumar, A.R., Montesano, R. and Saffiotti, U.: Aromatic amines carcinogenicity in hamsters. Proc Am Assoc. Cancer Res., 10, 78, 1969
  15. Lee, J.H., Shin, H.S., Kim, Y.H., Roh, J.H. and Lee, B.G. : A study on biomarker for bladder cancer screening workers employed for dyes and pigments manufacturing industries by GC/MS-SIM and 32P-postlabeling method, Kor. J. Env. Hlth. Soc., 26(1), 36-44. 2000