Abstract: A method was developed for the simultaneous determination of 3 Fluoroquinolones( FQs)( enrofloxacin,ciprofloxacin,ofloxacin) residues including enrofloxacin,ciprofloxacin,ofloxacin in seafood samples by new Qu ECh ERS and Enzyme- linked Immunosorbent Assay( ELISA). The approach involved the extraction of sample with acetonitrile containing 1%( v / v) acetic acid,and the supernatant liquid was directly purified with C18,meanwhile,was used for the precipitation of protein. The method was proved to be sensitive,rapid,precise,economical and applied. The calibration curve should be virtually linear in the range of 0.107 ~ 177.896 ng / m L,the limit of detection( LOD) was 0.0087μg / kg,and IC50 value was 4.34 ng / m L. Average recovery rate was 94.10% and93.70% separately in same batch and different batch,coefficient of variation was 4.09% ~8.41% and 2.78% ~7.99%separately in same batch and different batch.The results showed good coefficient with HPLC( R2= 0.9897),which proved to be suitable for the screening of seafood samples for the presence of FQs.