Simultaneous determination of eugenol, isoeugenol and methyleugenol in fish fillet using gas chromatography coupled to tandem mass spectrometry

•The fish anesthetics of eugenol, isoeugenol‚ and methyleugenol in fish fillet were simultaneously determined using GC–MS/MS.•Only three simple steps for sample preparation.•Be free of such steps as rotary evaporation and nitrogen blowing in sample preparation procedure.

Gas chromatography (GC) coupled with triple quadrupole tandem mass spectrometry (MS/MS) operated in electron ionization mode (EI) has been shown to have advantages in the trace analysis of chemical compounds. Employing the instrument, a method has been built to simultaneously determine eugenol, isoeugenol‚ and methyleugenol, which have been widely used as fish anesthetic, in the fish fillet. Procedure for the sample preparation was achieved by using hexane extraction followed by phenyl solid phase extraction (SPE) cleanup, which was free of such steps as rotary evaporation and nitrogen blowing by taking the volatility of eugenol and its isomers into consideration. The method was validated by conducting recovery studies on fortified fish fillet samples at four concentrations. The linearity in the range of 5–500 μg·L−1 was forced through the origin giving a coefficient of determination (r2) greater than 0.9982. Limits of detection (LODs) for eugenol, isoeugenol‚ and methyleugenol were 0.4, 1.2‚ and 0.2 μg·kg−1, respectively. The limits of quantification (LOQs) were 1.2, 4‚ and 0.7 μg·kg−1 for eugenol, isoeugenol‚ and methyleugenol, respectively. The recoveries for eugenol and its isomers ranged from 76.4 to 99.9% with relative standard deviations (RSD) in a range from 2.18 to 15.5%. This method is quick, simple and suitable for determining the residues of eugenol, isoeugenol and methyleugenol simultaneously in batch samples of fish fillet.