In vitro metabolism of (S)-(-)-[2'-14C]nicotine, using various tissue preparations of marmoset

The nicotine metabolite profile produced by marmoset liver, lung and kidney preparations was investigated after 30 minutes incubation of (S)-(-)-[2'-14C]nicotine. Cation-exchange high performance liquid radiochromatography was employed to separate and quantify nicotine and its metabolites. Cotinine-N-oxide (CNO, 0.7%), 3'-hydroxy-cotinine (3'-OH-C, 0.2%), norcotinine (NORC, 0.9%) and nornicotine (NORN, 0.4%) were formed in the incubates of marmoset lung homogenates; when marmoset kidney homogenates were used, CNO, 0.4%; 3'-OH-C, 0.2%; NORC, 0.7%; NORN, 0.7%; and cotinine (COT, 0.4%) were detected in the incubates. These nicotine metabolites constituted only approximately 2.2% and 2.4% of the original nicotine substrate used by lung and kidney homogenates respectively. When marmoset hepatic homogenates and microsomes were used, both COT and NORN were detected as the major nicotine metabolites. In addition, traces of CNO and 3'-OH-C were also detected in both incubates. The amounts of COT (6.4%) and NORN (1.8%) in the hepatic homogenates were approximately twice that of those formed by hepatic microsomes (3.8% and 0.9%, respectively). Nicotine-1'-N-oxide (NNO, 1.1%) was only detected in the latter preparation. Under the experimental conditions, these nicotine metabolites constituted only 8.2% and 5.8% of the substrate nicotine used in the respective incubates. The present results showed that both primary C-oxidation pathways, i.e. cotinine formation and N-demethylation of nicotine, occurred in the lung, kidney and liver of marmoset in vitro. However, N-oxidation of nicotine was only observed when a marmoset hepatic microsomal preparation was used.