Isolation and identification of non-chlorinated phenylbenzotriazole (non-ClPBTA)-type mutagens in the Ho River in Shizuoka Prefecture, Japan.
We previously identified 2-[2-(acetylamino)-4-amino-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA) congeners as major mutagens in water concentrates from several rivers that flow in three different areas, i.e. Kyoto, Aichi, and Fukui Prefectures, in Japan. In synthesis studies, these PBTAs were shown to be formed from corresponding dinitrophenylazo dyes via non-chlorinated derivatives (non-ClPBTAs). However, only non-ClPBTA-1, i.e. 2-[2-(acetylamino)-4-[bis(2-methoxyethyl)amino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole, had been detected as a minor contaminant in the Nishitakase River in Kyoto. In this study, analysis of mutagens in water concentrate from the Ho River, which flows through an area with a textile dyeing industry in Shizuoka Prefecture, Japan, allowed the isolation of four compounds (I, II, III, and IV). These four mutagens were identified as 2-[2-(acetylamino)-4-[N-(2-cyanoethyl)ethylamino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-2), 2-[2-(acetylamino)-4-[(2-hydroxyethyl)amino]-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-3), 2-(2-acetylamino-4-amino-5-methoxyphenyl)-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-4), and 2-[2-(acetylamino)-4-(diethylamino)-5-methoxyphenyl]-6-amino-4-bromo-2H-benzotriazole (non-ClPBTA-7) by spectral data and co-chromatography using synthesized standards. Non-ClPBTA-3 and -7 were highly mutagenic in Salmonella typhimurium YG1024, inducing 159,000 and 178,000 revertants/microg, respectively, in the presence of S9 mix. Like PBTAs, non-ClPBTAs might have been produced from azo dyes during industrial processes in dyeing factories and released into rivers.
Watanabe
Tetsushi
T
Department of Public Health, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan. watanabe@mb.kyoto-phu.ac.jp
Ohba
Hiroshi
H
Asanoma
Masaharu
M
Hasei
Tomohiro
T
Takamura
Takeji
T
Terao
Yoshiyasu
Y
Shiozawa
Tatsushi
T
Hirayama
Teruhisa
T
Wakabayashi
Keiji
K
Nukaya
Haruo
H
D002851:Chromatography, High Pressure Liquid; D004396:Coloring Agents / Q000737:chemistry / Q000302:isolation & purification / Q000633:toxicity; D005618:Fresh Water / Q000737:chemistry; D007220:Industrial Waste / Q000009:adverse effects; D007564:Japan; D009152:Mutagenicity Tests; D009153:Mutagens / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics; D021241:Spectrometry, Mass, Electrospray Ionization; D013784:Textiles; D014230:Triazoles / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity; D014874:Water Pollutants, Chemical / Q000302:isolation & purification* / Q000633:toxicity
D004396:Coloring Agents; D007220:Industrial Waste; D009153:Mutagens; D014230:Triazoles; D014874:Water Pollutants, Chemical