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The tobacco alkaloids: nicotine, nornicotine, cotinine, anabasine, their protonated forms, and salts were analyzed by means of
Raman spectroscopy supported by the density functional theory/B3LYP/aug-cc-pVDZ calculations. The analyses were per-
formed based on Raman marker bands of neutral, monoprotonated, and diprotonated forms of tobacco alkaloids because
in different surroundings various forms have been suggested to either dominate or to coexist. The form and distribution of
nicotine directly in a plant and in phytopharmaceutical products were investigated by
in situ
Raman mapping. For the
fi
rst
time, the Raman optical activity spectrum of (
)-nicotine in aqueous solution was measured and interpreted by means of
the density functional theory calculations. The study provides a clear evidence that Raman spectroscopy techniques are pow-
erful in ef
fi
cient quality control and forensic and bioanalytical analyses of tobacco alkaloids.