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배문주; 노재훈; 조영봉; 김춘성; 전미령; 김치년

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자료유형: 학술저널

저자정보: 배문주 (연세대학교 의과대학 산업보건연구소) 노재훈 (연세대학교 의과대학 산업보건연구소) 조영봉 (연세대학교 보건과학대학 산업환경학과) 김춘성 (연세대학교 의과대학 산업보건연구소) 전미령 (연세대학교 의과대학 산업보건연구소) 김치년 (연세대학교 의과대학 산업보건연구소)

저널정보: 한국산업보건학회 (구 한국산업위생학회) 한국산업보건학회지 한국산업보건학회지 제6권 제1호

발행연도: 1996.1

수록면: 28 - 37 (10page)

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Benzidine, an aromatic amine used primarily in the manufacture of azo dyes, is recognized as a urinary bladder carcinogen in humans. In rats, mice, and hamsters, chronic exposure to benzidine resulted in tumors of the liver. The present study was undertaken to suggest analyzing the metabolites of benzidine with the optimal condition, identify the metabolites of benzidine, and observe time variance of the metabolites in the isolated perfusated rat liver. N-acetylbenzidine was synthesized by acetylation of benzidine with acetic anhydride and separated by thin layer chromatography(TLC) and high performance liquid chromatography(HPLC). To analysis benzidine and the metabolites of benzidine, HPLC operating condition has been optimized by means of preliminary experiment. The mobile phase consisted of acetonitrile(37%) in phosphate buffer, flow rate maintained at 1.0 ml/min. Optimal detective conditions were electrochemicaldetector(ECD) at 0.75 V for benzidine and N-acetylbenzidine and ultravioletdetector(UVD) at 287 nm for N,N'-diacetylbenzidine. The separation system was composed of a guard column and a separation column(Polymer C18, $4.6{\times}250cm$) at a temparature of $40^{\circ}C$. The perfusion system was equilibrated for 30 minutes before addition of benzidine to the perfusate. Samples of the perfusate were collected at time intervals(0, 10, 20, 30, 60, 90, 120 min) during the 2 hour perfusion. Before analyzing samples by HPLC/ECD/UVD, samples had been treated with sep-pak. Samples of perfusate analyzed by HPLC/ECD/UVD and the metabolites of benzidine in the isolated perfused rat liver were N-acetylbenzidine and N,N'-diacetylbenzidine. Benzidine metabolized over 60% during the initial 30 minutes of perfusion, extensively by 1 hour, and was undetectable in the perfusate. N-acetylbenzidine increased by 30 minutes of perfusion, declined. N,N'-diacetylbenzidine increased the 0-90 minutes period, remained constant during the 90-120 minutes period.

#Isolated liver perfusion #High Performance Liquid Chromatography #Electrochemicaldetector #Ultravioletdetector #Benzidine #N-acetylbenzidine #N,N'-diacetylbenzidine

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