• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.10 no.1
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• pp.1-17
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• 2000

Carcinogen-DNA adduct analysis has potential for biomonitoring the earliest effects of exposure to many chemical carcinogens. They are the covalent reaction products of electrophiles and nucleophilic sites on DNA and the initial damage to DNA induced by many carcinogens. So many researchers begin to use them as biomarker for monitoring the earliest exposure of carcinogens and develop the effective analytical techniques about them. Randerath, Gupta and coworkers(1981, 1982) has also developed a $^{32}P$-postlabelling method as one among them. A major project for biomonitoring workers with carcinogen-DNA adducts is to develop non-invasive samples instead of tissues of target organs such as baldder and lung. This study use the exfoliated urothelial cells in urine for examine benzidine-DNA adducts. The content of exfoliated urothelial cells is not enough to significantly measure DNA content with spectrophotometer, and require the another way. So firstly washing the collected cells with PBS and 70% ethanol and centrifuge them for removing the crystals in urine, which block the isolation of DNA adducts. And then, measure the total nucleotide after $^{32}P$-postlabelling for calculating RAL. $[{\gamma}-^{32}P]ATP$ using for $^{32}P$-postlabelling, can synthesize with $[^{32}P]H_3PO_4$, and reagent and enzyme mixture (RM, EM), which is very economic in case of requiring a lot of them. Chromatography was composed of two steps. First step was to separate adduct ones from unadducted nucleotide, and secondary step was separate each adduct, which were performed with 4 kinds of solvents and different directions on TLC. With this procedure, we measure the DNA adducts in exfoliated urothelial cells of workers who were employed in benzidine and benzidine-dye company. RAL of adducts were $89.0{\times}10^7$ and $57.0{\times}10^7$ in them. In conclusion, we can significantly measure the DNA adduct in exfoliated urothelial cells by using the above $^{32}P$-postlabelling procedures, and use them to be biomonitoring workers who exposed carcinogens.

• Journal of Environmental Health Sciences
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• v.29 no.4
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• pp.21-26
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• 2003

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.