Abstract: The presence of sulfonamides, quinolones and macrolides antibiotic residues in environment is a major concern for both aquatic ecosystems and human health, particularly because they can induce drug-resistant bacteria and resistance genes. An efficient method was developed for the determination of sulfonamides, quinolones and macrolides antibiotics in sediment from aquaculture environment by QuEChERS (quick, easy, cheap, effective, rigged and safe)-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The target analytes in sediments were ultrasonically extracted with 20 mL phosphate buffer solution and acetonitrile mixture (1∶1,V/V) and salted out with sodium chloride (NaCl), followed by 0.15 g Na₂EDTA complexation. After centrifugation, the supernatant extract was cleaned up with magnesium sulfate (MgSO₄), primary secondary amine (PSA), C18 sorbent and graphitized carbon black. The analytes were separated on an Ultimate XB-C18 column (2.1 mm×150 mm×5 μm) by gradient elution with 4 mmol/L ammonium acetate solution containing 0.1% formic acid-0.1% formic acid methanol as mobile phase. The formic acid played a key role in mobile phase as modifier to improve the ionization efficiency of antibiotics. The column temperature was 40 ℃ and the flow rate was 0.3 mL/min. The analytes was performed using electrospray ionization (ESI) in positive ion mode and multiple reaction monitoring (MRM), quantified with external standard method. In order to achieve more accurate quantitation, a matrix-matched curve was used to eliminate the matrix effects. The results showed that the calibration curves are linear (R²>0.99) over a concentration range of 1.0-200 μg/L for 26 antibiotics. The limits of quantification are 1.2 μg/kg for all antibiotics. At three spiked levels, the average recoveries ranged from 60.4% to 113% with the relative standard deviations (RSDs) of 1.5%-13.6%. This method has high sensitivity, good reproducibility and wide applicability, which is suitable for rapid screening and detection of 26 antibiotics. Sulfonamides, quinolones and macrolides antibiotics were determined in 20 sediment samples collected from aquaculture ponds in southern Fujian Province. Sulfamerazine, sulfamethazine, sulfamethoxazole, sulfadiazine, erythromycin, roxithromycin and clarithromycin were detected, and the detection rates of sulfadiazine, sulfamethazine and sulfamethoxazole were the highest.