HPLC analysis of in vivo metabolites of 4-nitrobenzoic acid [(5-nitro-2-thiopheneyl)methylene]hydrazide in rats

Koçyiǧit-Kaymakçioǧlu B., Ünsalan S., Küçükgüzel Ş. G., Şener G., Rollas S.

European Journal of Drug Metabolism and Pharmacokinetics, vol.32, no.4, pp.197-203, 2007 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 32 Issue: 4
  • Publication Date: 2007
  • Doi Number: 10.1007/bf03191004
  • Journal Name: European Journal of Drug Metabolism and Pharmacokinetics
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.197-203
  • Keywords: hydrazone, in vivo metabolism, diode array detection
  • Istanbul Medipol University Affiliated: No


The in vivo metabolism of 4-nitrobenzoic acid [(5-nitro-2-thiopheneyl) methylene]hydrazide (substrate), a model that represents hydrazide hydrazone compounds, was investigated in the rat. The metabolites were monitored in rat plasma at certain time intervals. The substrate was dissolved in dimethylsulfoxide (DMSO)/water (1:4) and administered intraperitoneally at dose of 100 mg/kg and 500 mg/kg. Blood samples were collected at 30 min, then at 1, 2, 4, 8, 12 and 24 h post-administration. The chromatographic separation of the substrate and its metabolites was performed on a Novapak C18 (Phenomenex) (150 mm x 4.6 mm i.d., 5-μm particle size) using a mobile phase consisting of phosphate buffer: acetonitrile (90:10, v/v) with a linear gradient system. From the biotransformation of this compound, 4-nitrobenzoic acid (M3) was identified together with the substrate, as evidenced by high pressure liquid chromatography (HPLC)-UV/ diode array detection (DAD).