Mechanisms for the emergence of chemical defense are of significant interest to chemical ecologists. In particular, acquired chemical defense through diet requires progressive development of resistance to chemical toxins to allow increased consumption thereof and/or active sequestration. Central to any study of these traits is the identification and quantitation of toxic compounds and their metabolites. To this end, we examined extracts of <i>D. melanogaster</i> flies raised on nicotine-containing media to assess uptake and metabolism. We identified nicotine and its oxidation product cotinine by gas chromatography-unit resolution mass spectrometry in an ion trap instrument. Using ultra high pressure liquid chromatography-high resolution mass spectrometry with an orbitrap instrument, we were able to identify compounds corresponding to both oxidation and glucuronidation of nicotine. Tandem mass spectrometry allowed for unambiguous identification of metabolites in the absence of analytical standards based on fragments obtained through higher energy collision induced dissociation (HCD). Our results are consistent with literature precedent in other organisms with regard to retention time and observed fragments, though a clear energy dependence on the fragments observed was noted. Optimization of sample preparation, chromatography and mass spectral parameters associated with this study will be presented as well as preliminary quantitative results of these metabolites in flies at various developmental stages.

Analysis of nicotine and metabolites in <i>Drosophila melanogaster</i> raised on nicotine-containing media

ACS Spring 2022

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Analysis of nicotine and metabolites in Drosophila melanogaster raised on nicotine-containing media