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Radioflow HPLC analysis of CYP2A13-mediated metabolism of nicotine Δ5′(1′)iminium ion on system I. CYP2A13 (3 pmol) was incubated with 20 μM [5-3H]nicotine Δ5′(1′)iminium ion (specific activity, 0.1 μCi/nmol) for 5 min (A) or 20 min (B). The retention times were confirmed by coelution with standards in each sample.
