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흰쥐의 적출 간 관류법을 이용한 벤지딘 대사에 관한 연구
조영봉,김치년,김춘성,배문주,노재훈,전미령 한국산업위생학회 1996 한국산업보건학회지 Vol.6 No.1
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 analysing 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 ㎖/min. Optimal detective conditions were electrochemicaldetector(ECD) at 0,75 V for benzidine and N-acetylbenzidne and ultravioletdetector(UVD) at 287 nm for N,N¹-diacetylbenzidine. The separation system was composed of a guard column and a separation column(Polymer C 18, 4.6 × 250㎝) at a temperature of 40℃. 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,N¹-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,N¹-diacetylbenzidine increased the 0-90 minutes period, remained constant during the 90-120 minutes period.
Synthesis and Characterization of N-Methylamidinoglycine : an Isomer of Creatine
조영봉,백운기,Young-Bong Cho,R. John Stedman,Woon Ki Paik Korean Chemical Society 1985 대한화학회지 Vol.29 No.4
N-Methylamidinoglycine, an isomer of creatine which was postulated to be formed enzymatically in vitro, has been synthesized by coupling glycine with N, S-dimethylthiopseudouronium iodide in a yield of approxirnately 60%. On heating in acidic solution, it was converted to a cyclized form (isocreatinine) in analogy with the conversion of creatine to creatinine(anhydrous form). Structures were confirmed by an elemental analysis and proton NMR spectroscopy. Further studies on their characteristics were compared with those of creatine and creatinine in regard to isoelectric points(pI), retardation coefficients(Rf) on thin layer chromatography, and elution profiles on amino acid analyzer. In order to facilitate the comparison, $^{14}C$-labeled creatine, creatinine, isocreatine and isocreatinine were also synthesized. 시험관내에서 효소에 의해 생성될 수 있을 것으로 생각되는 N-Methylamidinoglycine(isocreatine)을 glycine과 N, S-dimethylthiopseudouronium iodide로 부터 약 60% 수득율로 합성하였고, isocreation의 산성수용액을 가열하여, creatine이 creatinine으로 탈수 고리화되는 것처럼, 고리화된 isocreatinine도 얻었다. 한편 이들 화합물에 대해 원소분석, nmr 스펙트럼, 박층 크로마토그피(Rf) 및 아미노산분석기에서의 elution rate도 검토하였으며, 등전점을 측정하기 위해서 $^{14}C$-creatine, $^{14}C$-creatinine, $^{14}C$-isocreatine 및 $^{14}C$-isocreatinine도 합성하였다.
적출 흰쥐 간 관류법에 의한 이황화탄소 대사에 관한 연구
조영봉,최홍순,노재훈,배문주 한국산업위생학회 1996 한국산업보건학회지 Vol.6 No.2
The purposes of this study are the identification and determination of metabolites in the isolated rat liver perfusate of carbon disulfide by two-dimentional thin-layer chromatography and high performance liquid chromatography for understanding the metabolism of carbon disulfide. 2-Thio-thiazolidine-4-carboxylic acid(TTCA) was synthesized by the reaction of carbon disulfide and cysteine, and confirmed by two-dimentional thin-layer chromatography, high performance liquid chromatography, UV spectroscopy, and IR spectroscopy. The absorbance of UV detector for the simultaneous determination of TTCA and thiocarbamide was 254 ㎚ although their maximum spectra were 273 ㎚ and 237 ㎚, respectively. Two kinds of the developing solvent in the two-dimentional thin-layer chromatography were 2-butanol : 80% HCOOH : H₂O(7 : 2 : 1) as the first developing solvent and 2-propanol : H₂O (4 : 1) as the second developing solvent. After perfusion of carbon disulfide (8274.23 umol), the amount of TTCA and thiocabamide of the perfusate(100 ㎕) were 12.02 - 16.43 umol and 5.25 - 8.l5㎍, respectively. The mean amount of them were 14.08 umol and 6.41 umo1 respectively, and the former was 2.20 times greater than the latter. For conforming the mechanism of formations of TTCA and thiocarbamide in vivo, we have to clarify whether the reactions between carbon disulfide and ammonia, ammonium salts, amides, cysteine, cystine, or proteins will be formed in vitro.
흰쥐에서 NG - Monomethyl - L - arginine 으로부터 methylamine 의 생성
조영봉,최홍순,김춘성,안영곤 한국산업위생학회 1996 한국산업보건학회지 Vol.6 No.1
After oral administration of 14C-labelled N^G-mono[methyl-14C]-L-arginine into rats, 38.2 % and 14.7 % of the administered radioactivity had been recovered in the urine and stool during 10 days. In the urine, 59.4 % of the radioactivity was recovered in the first 24-hours and used for the identification of the formation of methylamine. The strong canon-exchange resin column chromatography showed 6.3 %, 7.4 %, 4.9 %, and 81.5 % of the distributions of radioactivity of the neutral, monomethylamine, basic, and uneluted portions, respectively. The radioactivity of monomethylamine portion reeluted into the column chromatography was 39.5 %. The radioactivities corresponding monomethylamine in the column chromatography, thin-layer chromatography, and thin-layer electrophoresis were 39.5 %, 37.3 %, and 28.8 % of the recovered radioactivity, respectively.
흰쥐에서 N<sup>G</sup>-Monomethyl-L-arginine으로부터 methylamine의 생성
조영봉,안영곤,최홍순,김춘성,Cho, Young Bong,Ahn, Young Kon,Choi, Hong Soon,Kim, Choon Sung 한국산업보건학회 1996 한국산업보건학회지 Vol.6 No.1
After oral administration of 14C-labelled $N^G$-mono[methyl-14C]-L-arginine into rats, 38.2 % and 14.7 % of the administered radioactivity bad been recovered in the urine and stool during 10 days. In the urine, 59.4 % of the radioactivity was recovered in the first 24-hours and used for the indentification of the formation of methylamine. The strong cation-exchange resin column chromatography showed 6.3 %, 7.4 %, 4.9 %, and 81.5 % of the distributions of radioactivity of the neutral, monomethylamine, basic, and uneluted portions, respectively. The radioactivity of monomethylamine portion reeluted into the column chromatography was 39.5 %. The radioactivities corresponding monomethylamine in the column chromatography, thin-layer chromatography, and thin-layer electrophoresis were 39.5 %, 37.3 %, and 28.8 % of the recovered radioactivity, respectively.