title: Identification of N2 as a metabolite of acetylhydrazine in the rat.
abstract: In the pathogenesis of isoniazid-induced hepatic injury, cytochrome P450-dependent metabolic activation of the metabolite, acetylhydrazine (AcHz), is the crucial step. Exhalation of [14C]-carbon dioxide has previously been used to quantify indirectly this pathway. In contrast, according to the current concept of AcHz bioactivation, molecular nitrogen is produced directly, but has not yet been identified. Here, we measured [15N]-nitrogen and 14CO2 exhalation, after the administration of [15N2]-[14C]-AcHz, in rats. Laser magnetic resonance (LMR) spectroscopy, a new sensitive and specific technique for the measurement of 15N and 14N in gas samples, was used. To demonstrate the involvement of cytochrome P450, rats were treated with phenobarbital (PB) or PB+cobalt(II) chloride (CoCl2) (n = 3 in each group). Time-dependent 15N2 exhalation differed significantly between treatment groups (p < 0.001). At 240 min, cumulative exhalation of 15N was 1.92 +/- 0.43% (mean +/- SE) of the dose in the control group, 2.53 +/- 0.23% in the PB group, and 1.00 +/- 0.15% in the PB+CoCl2 group (p < 0.05 compared to controls, p < 0.01 compared to PB). Cumulative exhalation of 14CO2 in 24 h ranged from 15.1 to 21.9%, with no significant difference between treatment groups. In conclusion, N2 is a metabolite of AcHz. N2 formation reflects the cytochrome P450-mediated activation of AcHz and can be used as an index of this pathway. Generally, LMR spectroscopy is valuable for monitoring any N2-liberating process in vivo.
authors.affiliation: Dr. Margarete Fischer-Bosch-Institut für Klinische Pharmakologie, Stuttgart, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Mörike
authors.forename: M
authors.initials: M
authors.lastname: Koch
authors.forename: P
authors.initials: P
authors.lastname: Fritz
authors.forename: W
authors.initials: W
authors.lastname: Urban
authors.forename: M
authors.initials: M
authors.lastname: Eichelbaum
mesh_terms: D000818:Animals; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D006834:Hydrazines / Q000493:pharmacokinetics* / Q000633:toxicity; D007834:Lasers; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D009584:Nitrogen / Q000378:metabolism*; D009587:Nitrogen Isotopes; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D006834:Hydrazines; D009587:Nitrogen Isotopes; D009569:Nitric Oxide; D003577:Cytochrome P-450 Enzyme System; D009584:Nitrogen; C014949:acetylhydrazine