• 한국환경보건학회지
• /
• 제29권2호
• /
• pp.50-55
• /
• 2003

3,3'-디클로로벤지딘(DCB)는 실험동물에 발암물질로 밝혀졌고, 사람에게 암을 유발시킬 수 있는 발암물질로 의심되고있다. 많은 연구자들이 사업장에서 DCB에 폭로된 근로자들을 대상으로 뇨중에 배설된 대사물질, 헤모글로빈 부가체, 그리고 암 발생율 등에 대하여 연구를 하고 있다. 이러한 연구를 하기 위해서는 표준물질로 되어 있는 DCB의 대사물질이 꼭 필요하다. 따라서 본 연구의 목적은 DCB를 이용하여 이들의 대사물질을 합성하여 표준물질로 사용코자 합니다. DCB는 벤젠. 에테르, 에탄올, 메탄올 등에 부분적으로 용해되지만, 구연산이 1 % 이하로 함유된 70% 아세트산, 피리딘, 0.1N NaOH와 톨로엔이 1:2로 섞인 혼합물, 20 mM TRIZA염으로 포화된 페놀 등에는 완전히 용해되기 때문에 본 연구에서는 DCB를 피리딘에 녹여서 사용하였다. DCB와 대사물질인 mono-acetyl-DCB 및 diacetyl-DCB는 가스크로마노그래피(GC/MS)로 분석하였고, 검출기는 NPD와 SIM를 사용하였다. DCB의 기본피크는 252 m/z이었고, mono-acetyl-DCB의 기본피크는 252와 294 m/z로 구성되어 있었으며. diacetyl-DCB의 기본피크는 252, 294, 336 m/z로 구성되어 있었다. Discetyl-DCB는 피리딘에 용해된 DCB에 염소아세틸를 충분히 적정하여 합성하였다. 이렇게 얻은 diacetyl-DCB의 순도는 98.7%이었다. 침전물위에 있는 용해물질 속에는 DCB. mono-acetyl-DCB, diacetyl-DCB가 함유되어 있었는데, 아세트산을 아세틸화를 조절하는 물질로 사용하여 DCB를 모두 아세틸화시키었고, diacetyl-DCB로부터 mono-acetyl-DCB를 분리하여 추출하였다. 추출된 mono-acetyl-DCB는 아세톤으로 세척하여 98.8%의 순도를 얻었다.

• 한국환경보건학회지
• /
• 제28권1호
• /
• pp.21-29
• /
• 2002

To identify and evaluate the dichlorobenzidine(DCB)-DNA adducts in liver cell and bladder epithelial cells by $^{32}$ P-postlabeling and GC/MS-SIM, we orally exposed the dichlorobenzidine(20mg/kh body wt./day) to male Sprague-Dawley rats(l85$\pm$10g) for 14 days. Two kinds of DCB-DNA adduct(A1 and A2) were found at the same site of thin layer chromatogram of $^{32}$ P-postlabeling method in liver cells and bladder epithelial cells. In liver cells, relative adduct labeling(RAL) $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 34.1$\pm$3.71 and 69.9$\pm$5.02, that of adduct A2 were 74.1$\pm$10.1 and 105.1$\pm$10.1 on 10 and 14 days after treatment, respectively. And in bladder epithelia cells, RAL $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 5.92$\pm$1.60 and 15.9$\pm$1.31, that of adduct A2 were 9.81$\pm$2.81 and 22.8$\pm$1.79 on 10 and 14 days after treatment, respectively. DCB metabolites formed DNA adducts were monoacetyl-dichlorobenzidine(acDCB) and diacetyl-dichlorobenzidine(di-acDCB), which was identify by gas chromatography/mass spectrometry-scan ionization mode(GC/MS-SIM), after hydrolysis of DCB-DNA adducts isolated from live cells and bladder epithelial cells. The base peak of acDCB were 252 and 294 m/z, and that of di-acDCB were 252, 294 and 336 m/z. In conclusion, the exposed DCB formed two kinds of DCB-DNA adduct, the proximate materials of that were acDCB and di-acDCB in liver and bladder epithelial cells. And the above GC/MS-SIM method was found the DCB-DNA adducts could be monitoring by gas chromatography.

• 한국환경보건학회:학술대회논문집
• /
• 한국환경보건학회 2002년도 춘계 국제 학술대회
• /
• pp.49-51
• /
• 2002

To identify and evaluate the dichlorobenzidine(DCB)-DNA adducts in liver cell and bladder epithelial cells by $^{32}$ P-postlabeling and GC/MS-SIM, we orally exposed the dichlorobenzidine (20mg/kh body wt.,/day)to male sprague-dawley rats for 14 days. Two kinds of DCB-DNA adduct were found at the same site of thin layer chromatogram of $^{32}$ P-postlabeling method in liver cells and bladder epithelial cells. In liver cells, relative adduct labeling(RAL) $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 34.1$\pm$3.71 and 69.9$\pm$5.02, that of adduct A2 were 74.1$\pm$10.1 and 105.1$\pm$10.1 on 10 and 14 days after treatment, respectively. And in bladder epithelia cells, RAL $\times$ 10$^{12}$ of DCB-DNA adduct A1 were 5.92$\pm$1.60 and 15.9$\pm$1.31, that of adduct A2 were 9,81$\pm$2.81 and 22.8$\pm$1.79 on 10 and 14 days after treatment, respectively. DCB metabolites formed DNA adducts were monoacetyl-dichlorobenzidine(acDCB) and diacety1-dichlorobenzidine(di-acDCB), which was identify by gas chromatography/mass spectrometry-scan ionization mode(GC/MS-SIM), along with hydrolysis, extraction and TFA(trifluoroacetyl anhyride) derivatization with DCB-DNA adducts isolated from live cells and bladder epithelial cells. The base peak of acDCB were 252 and 294 m/z, and that of di-acDCB were 252, 294 and 336 m/z. In conclusion, the exposed DCB formed two kinds of DCB-DNA adduct, the proximate materials of that were acDCB and di-acDCB in liver and bladder epithlial cells. And the above GC/MS-SIM method was found the DCB-DNA adducts could be monitoring by gas chromatography.