Fig. 3.

Chromatograms are the result of product Total Ion Current (TIC) or multiple reaction monitoring (MRM) of fenbendazole (FBZ) parental drug and its metabolites identified in the supernatant of cultures of C. elegans benzimidazole resistant allele CB3474 ben-1(e1880)III (shown in green) or of adult H. contortus drug sensitive MHco3(ISE) allele (shown in orange). The first (top) chromatogram corresponds to the culture containing benzimidazole but no nematodes, the second chromatogram corresponds to the culture containing either C. elegans or H. contortus respectively, exposed to the benzimidazole, and the third chromatogram corresponds to the control culture containing C. elegans or H. contortus but with no benzimidazole. C. elegans was exposed to 56.5 μM BZ for 3 days, and H. contortus was exposed to 10 μM BZ for 16 h respectively. A. TIC scan of C. elegans exposed to fenbendazole (FBZ), B. TIC scan of H. contortus exposed to fenbendazole (FBZ), C. An extracted-ion chromatogram (EIC) of C. elegans exposed to fenbendazole (FBZ), D. MRM scan of H. contortus exposed to fenbendazole (FBZ). Metabolites and their LC-MS/MS fragmentation patterns identified in supernatant of the FBZ-treated C. elegans or adult H. contortus cultures. These fragmentation patterns were extracted from a Total Ion Current (TIC) or multiple reaction monitoring (MRM) and resulted from a fragmentor setting of 150 V and a collision energy of 30eV. E. Metabolite 5b from C. elegans and its LC-MS/MS fragmentation pattern, F. Metabolite 6 and its LC-MS/MS fragmentation pattern, G. Metabolite 5b from H. contortus and its LC-MS/MS fragmentation pattern, H. Metabolite 7 and its LC-MS/MS fragmentation pattern. I. A schematic summary of FBZ oxidative biotransformation is shown. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article).