Development and Application of a Rapid Screening SPE-LC-QTOF Method for the Quantification of 14 Anesthetics in Aquatic Products

Anesthetics are widely applied in cultivation and transportation of aquatic products to reduce the metabolism of fish and avoid some stress reactions. The overdose of using anesthetics or the occurrence of their residues in the aquatic products may negatively affect human health after consumption. A rapid screening procedure and a new analytical method for the simultaneous determination of 14 anesthetics was established in this study. The compounds were extracted from fish and shrimp matrices by sodium acetate buffer solution, cleaned up via solid-phase extraction, and then were analyzed using an ultrahigh-performance liquid chromatography system coupled with a tandem quadrupole-time-of-flight mass spectrometer (UPLC-Q-TOF–MS) in a positive-ion mode. Methodological verification analyses, including linearity, precision, recovery, limit of detection (LOD), and limit of quantification (LOQ), were conducted to validate the method. The correlation coefficient of the calibration curves of each anesthetic exceeded 0.99. The average recoveries of this analysis method varied from 72.36 to 111.27% with relative standard deviations less than 5%. The detection limit of anesthetics reached 0.3–0.7 μg/kg. Agilent Mass Hunter Personal Compound Database Library (PCDL) Manager was used to establish a database of 14 anesthetics and 8 isotope internal standards, which enabled a quick screening and analysis without standard substances. The results of the spiked blank samples demonstrated reliability of the screening method with mass deviations less than 5 ppm and the scores determined by relative molecular mass, retention time, isotope ratio, and distribution all greater than 85. In addition, according to the fragmentation information of mass spectrometry, the fragmentation rules of some anesthetics were summarized. This method was validated with reliable results in quantitative and qualitative analysis and could be used as regulatory measures for anesthetics in aquatic products.