title: High-pressure liquid chromatographic analysis for identification of in vitro and in vivo metabolites of 4-phenethyl-5-[4-(1-(2-hydroxyethyl)-3,5-dimethyl-4-pyrazolylazo)phenyl]-2,4-dihydro-3H-1,2,4-triazole-3-thione in rats.
abstract: All azo colorants whose metabolism can liberate a carcinogenic arylamine, are suspected of having carcinogenic potential. Therefore, a new azo compound 4-phenethyl-5-[4-(1-(2-hydroxyethyl)-3,5-dimethyl-4-pyrazolylazo)phenyl]-2,4-dihydro-3H-1,2,4-triazole-3-thione (substrate) was prepared to investigate its in vitro and in vivo biotransformation in rats by HPLC. Chromatographic separation of substrate and its metabolites was performed using a Chromasil C(18) column. The mobile phase consisted of acetonitrile and water in a linear gradient system. From the biotransformation of this compound, the reduction metabolite 4-(2-phenethyl)-5-(4-aminophenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was identified by comparing it to reference standard by HPLC-DAD. In the in vivo study, identification of the unknown peak which was the N-acetylation metabolite was confirmed by LC-MS spectrometry. Besides this, the azo compound was reduced to its corresponding amine in intestinal and cytosolic parts. In addition, oxidation of the methyl group and the phenyl ring, and reduction of azo group to hydrazo were identified in the cytosolic part using LC-MS.
authors.affiliation: Department of Pharmaceutical Chemistry, Marmara University, Facult of Pharmacy, Tibbiye Street 49, Haydarpasa, Istanbul, Turkey.
authors.forename: Bedia Koçyiğit
authors.initials: BK
authors.lastname: Kaymakçioğlu
authors.forename: Emine Elçin
authors.initials: EE
authors.lastname: Oruç
authors.forename: Seda
authors.initials: S
authors.lastname: Unsalan
authors.forename: Levent
authors.initials: L
authors.lastname: Kabasakal
authors.forename: Sevim
authors.initials: S
authors.lastname: Rollas
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002853:Chromatography, Liquid; D003600:Cytosol / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013871:Thiones / Q000032:analysis* / Q000378:metabolism; D014230:Triazoles / Q000032:analysis* / Q000378:metabolism
chemical_list: C509053:4-phenethyl-5-(4-(1-(2-hydroxyethyl)-3,5-dimethyl-4-pyrazolylazo)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione; D013871:Thiones; D014230:Triazoles