<p>An online turbulent flow chromatography-liquid chromatography-tandem mass spectrometry (online TFC-LC-MS/MS) method was created to quantify 14 sedatives and anesthetics residues in aquaculture products. Sample preparation, including extraction with acetonitrile with 1% ammonium hydroxide solution (v/v), concentration, and filtration, was followed by online TFC cleanup. The developed method was validated in accordance with the guidelines of SANTE/11312/2021. The matrix-match calibration standard curves developed for quantitative analysis had linear regression coefficients (<i>R</i>^<i>2</i>) higher than 0.9944. The LODs and LOQs of sedatives and anesthetics in aquaculture products ranged from 0.02 to 0.03 µg/kg and 0.05 to 0.10 µg/kg, respectively. The recovery rates at various spiking levels ranged from 79.4% to 119.5%. Both intra-day and inter-day RSDs were lower than 19.3%. Method accuracy was further verified using certified reference materials. These results indicate that the established approach is a sensitive, rapid, reliable, and promising tool for monitoring sedative and anesthetic residues in aquaculture products.</p>