[Determination of 5 kinds of fish anesthetics residues in fish by ultra-high performance liquid chromatography-tandem mass spectrometry].

OBJECTIVE: A method for the simultaneous determination of 5 kinds of fish anesthetics residues in fish has been developed by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Eugenol, methyl-eugenol, methyl-isoeugenol, acetyl-isoeugenol and tricaine methanesulfonate(MS-222) were concerned. METHODS: After homogenization fish samples were extracted by acetonitrile-water(80↿0, V/V), purified by Oasis PRiME HLB solid-phase extraction column. Then after centrifuged and concentrated, the samples were separated by Waters ACQUITY UPLC BEH Phenyl column(2. 1 mm×100 mm, 1. 7 μm). The detection was confirmed and quantified by mass spectrum of triple quadrupole in the multiple reaction monitoring(MRM) mode. RESULTS: The calibration curves showed good linearity in each range with correlation coefficients greater than 0. 995. Three levels spiked recovery experiments were carried out using blank fish mud extraction as substrate, the recoveries ranged from 72. 6% to 106. 0%, the relative standard deviations(RSDs) ranged from 2. 2% to 20. 1%(n=6). The qualitative limits of detections(S/N>3) were 0. 14-0. 30 μg/kg and the quantitative limits(S/N>10) were 0. 5-1. 0 μg/kg. CONSLUSION: The method is simple and easy to operate, with less organic reagent, high sensitivity and good stability. The isomers of methyl eugenol and methyl isoeugenol were successfully separated. It is suitable for the detection of 5 kinds of fish anesthetics in fish.