Simultaneous determination of multi-class antibiotic residues in aquatic products based on rapid pretreatment and sensitive immunochromatographic assay

• A quick pretreatment protocol for multi-class antibiotic residues from aquatic products was introduced. • The overall pretreatment time was reduced by 50%. • Simultaneous determination of multi-class antibiotic residues based on rapid and sensitive ICA. • Method validated in actual samples and certified by LC-MS/MS. The extensive use of multi-class antibiotics to bolster disease prevention in aquaculture has precipitated the issue of multi-class antibiotic residues in aquatic products. Most reported rapid detection methods predominantly target a limited scope of analytes, typically one or two, rendering them inadequate for comprehensive residue analysis. The unique physicochemical properties and co-extraction efficiency of these antibiotics further complicate their simultaneous detection. For this, a novel pretreatment protocol to facilitate the concurrent extraction of 3-amino-2-oxazolidinone (AOZ), semicarbazide (SEM), chloramphenicol (CAP), and malachite green (MG) from aquatic products was introduced. Under optimal conditions, the derivative time was decreased from 20 to 5 min, and the overall pretreatment time was reduced by 50 %, compared with those reported in previous studies. Moreover, a single pretreatment realized the simultaneous extraction of four illegal antibiotic residues in on-site detection, saving time and minimizing the sample required in aquatic product detection. Based on this pretreatment process, immunochromatographic assays (ICAs) were developed to determine four antibiotics simultaneously. The ICAs demonstrated high recovery rates, precision, and selectivity. The sensitivities of ICAs are all below the national standard required or recommended level. This developed method represents a significant advancement in screening banned antibiotics in aquatic products, offering a streamlined solution to a complex analytical challenge at rapid screenings.