The rise of aquaculture has necessitated the use of antibiotics and other agents in these densely populated and often closed environments. An enantioselective depletion study of four chiral 4-hydroxypraziquantel metabolites (4-OH-PZQ) in perch, tilapia, and ricefield-eel muscles was done using a simple, sensitive, and reliable liquid-chromatography−tandem-massspectrometry (LC-MS/MS) method. These metabolites result from the uptake of the drug praziquantel (PZQ), which is used in aquaculture. A novel strategy of using a C 18 short column in tandem with a chiral hydroxypropyl-β-cyclodextrin superficially porous particle (CDShell-RSP) column produced the optimal separation for both the enantiomers and diastereoisomers of 4-OH-PZQ. The method was linear over the concentration range of 1−250 μg L −1 (r 2 ≥ 0.99) for R-trans-4-OH-PZQ, S-trans-4-OH-PZQ, R-cis-4-OH-PZQ, and S-cis-4-OH-PZQ. The average recoveries of four analytes at three spiked levels of 1, 10, and 100 μg kg −1 ranged from 84.2 to 93.1%, and the intraday and interday relative standard deviations were less than 7.9%. The limits of quantification of the four 4-OH-PZQ metabolites in perch-, tilapia-, and ricefield-eel-muscle matrices were 1.0 μg kg −1 . The method was utilized to monitor the depletion of trans-and cis-4-OH-PZQ enantiomers in perch, tilapia, and ricefield-eel muscle following oral administration (medicine bath for ricefield eel). Species-specific differences in the PZQ metabolism of isomers were observed. In addition, new metabolites of PZQ were observed: 3-hydroxypraziquantel diastereomers.
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