Analysis of Sea Cucumber (Stichopus japonicas) Enrofloxacin Metabolites by Liquid Chromatography Coupled with Quadrupole-linear Ion Trap Mass Spectrometry
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Abstract:
Homogenized sea cucumber (Stichopus japonicas) samples were collected after incubating in medicated enrofloxacin bath. Metabolites were extracted from samples using acidified acetonitrile to identify the major metabolites of enrofloxacin in sea cucumber. The extracts were concentrated and purified using hexane, and analyzed by liquid chromatography coupled with quadrupole-linear ion trap mass spectrometry. Multiple reaction monitoring-information dependent acquisition-enhanced product ion (MRM-IDA-EPI) scanning was performed using an electrospray ionization source operated in positive mode to elucidate the structures of the major sea cucumber enrofloxacin metabolites. Ten potential metabolites were found after 6 h of drug administration: an isomerization product of enrofloxacin (M3); a deethylation product, ciprofloxacin (M1), and its isomer (M2); a hydrogenation reduction product and its isomers (M4, M5, and M6); a hydroxylation product and its isomer (M7 and M8); and an oxygenation product and its isomers (M9 and M10). The peak areas of sea cucumber enrofloxacin metabolites M2~M5 were higher than that of ciprofloxacin (M1). Metabolism of enrofloxacin primarily occurred by deethylation and hydrogenation reduction reactions, and M2 and M4 were the major enrofloxacin metabolites identified in sea cucumber.
