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Figure 1.

Nicotine metabolism.

Following nicotine uptake in the body, nicotine is metabolized to six primary metabolites (cotinine, hydroxycotinine, norcotinine, nornicotine, cotinine oxide, and nicotine oxide). Nicotine, cotinine, and trans-3′-hydroxycotinine are subsequently glucuronidated at significant rates.

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Figure 1 Expand

Table 1.

Standard Concentrations and Reportable Ranges (sample volume 200 µL urine–1∶5 dilution factor).

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Table 2.

HPLC Gradient Elution Table.

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Table 2 Expand

Figure 2.

Representative chromatograms.

(A) Standards Analysis (cotinine, 200 ng/mL); (B) Smoker Urine Sample (“free” cotinine, 14.1 ng/mL); (C) Smoker Urine Sample (“free” cotinine, 2767 ng/mL); (D) Smoker Urine Sample (“total” cotinine, 4195 ng/mL). Abbreviations: Cotinine-oxide (COX); Nicotine-oxide NOX); Hydroxycotine (HCT); Norcotinine (NCT); Cotinine (COT); Nornicotine (NNC); Anatabine (ANT); Anabasine (ANB); Nicotine (NIC). The fourth letter “T” in the abbreviations in Figure 2(D) represents the “total” concentrations for measured analytes.

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Figure 2 Expand

Table 3.

Accuracy and Precision from the Repetitive Analysis of Five Fortified Urine Pools.

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Table 4.

Limits of Detection (LOD) for “free” and ”total” analytes.

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Table 5.

Comparison of standard curves generated by urine or enzyme standards to water standard curve.

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Table 6.

Dilution influence on ion suppression. Abbreviations are according to Figure 2.

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Table 7.

The molar percent each analyte contributed to the combined concentrations of nicotine and 6 nicotine metabolites in this study (n = 94).

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Table 7 Expand