Objective To determine the residual levels of fish sedatives and quinolones residues in commercially freshwater fish and evaluate their dietary exposure risk. Methods A total of 77 batches of freshwater fish were collected in 8 administrative regions of Wuhan City. High performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) was used to determine the content of fishing anesthetics (MS-222, benzocaine, eugenol) and quinolone antibiotics (enrofloxacin, ciprofloxacin, ofloxacin, levofloxacin, levofloxacin, norfloxacin), and the dietary exposure risk of each compound was analyzed using a quantitative analysis method. Results MS-222 and benzocaine were not detected in all samples, and 48 batches of eugenol were detected with a content range of 2.59-1927.30 μg/kg, with a failure rate of 29.8%. The dietary exposure risk values for all age groups were less than 0.4%, indicating low dietary risk. All samples did not detect peofloxacin and norfloxacin. Enrofloxacin was detected in 34 batches with a content range of 1.74-5585.00 μg/kg, and ofloxacin was detected in 2 batches with a content range of 36.30-65.60 μg/kg, the overall failure rate was 18.2%, and the dietary exposure risk of all age groups was less than 8.00%. According to species analysis, the content range of enrofloxacin in loaches and yellow catfish was as follows: 2.74-5585.00 μg/kg and 6.82-1014.00 μg/kg, with dietary risks of 55.4% and 9.0%, respectively. Conclusion The fishery anesthetic used in this region is eugenol, which is characterized by high detection rate and large residual amount, and requires the development of normative documents to guide its use. Enrofloxacin, as the main quinolone antibiotic used in the region, has two high-risk species: Loach and yellow catfish. Market regulatory authorities should focus on the supervision of these two species.