Abstract:In order to detect the metabolic residues of eugenol anesthetics during the transportation of fresh aquatic products, a high performance liquid chromatography (HPLC) method for the simultaneous determination of six eugenol compounds in the muscle tissue of catfish (Ietalurus punetaus) was developed. The freeze-dried fish muscle tissue was ultrasonically extracted with a mixture of 15 mL methanol and acetonitrile (V/V, 2:1), purified by 100 mg of hydrophobic gas phase nano-silica, and filtered. The mobile phase was a mixture of methanol and 0.1% acetic acid solution (V/V, 58:42) with a flow rate of 0.8 mL/min and ultraviolet detection wavelength at 272 nm, the column temperature was 35 ℃, the injection volume was 10 μL, and the external standard method was developed. The linear relationship of six eugenol compounds was good in the range of 0.02~100.0 ug/mL with a linear regression coefficient more than 0.999. The average intra-recovery was between 50.52% and 97.98% and the relative standard deviation was between 1.25 and 14.56% at three different addition levels. The stability and repeatability of the method were good. The detection limit of the method is 0.026~0.121 mg/kg, which is lower than the residue limit standard of eugenol in Japan. It can be used for the detection of various eugenol compounds in fish samples in batches.
