QuEChERS and 96-well plate solid phase extraction for determination of vancomycin and norvancomycin in fish meat by UPLC–MS/MS

•Vancomycin and norvancomycin are antibiotics indiscriminately used in aquaculture.•QuEChERS/96-well SPE method was used to extract the drugs in fish meat.•The drugs were analyzed by UPLC–MS/MS with validation in linearity, precision, etc.•This method meets the daily monitoring requirements for vancomycin and norvancomycin. Vancomycin and norvancomycin are glycopeptide antibiotics for gram-positive bacteria infection, but indiscriminately used in aquaculture. In this study, a QuEChERS (quick, easy, cheap, effective, rugged, and safe)/96-well solid-phase extraction (SPE) plate method was used to extract vancomycin and norvancomycin in fish meat samples, and the drugs were further analyzed by ultra-performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS). The parameters, such as the sorbent of cation exchange resin, the proportion of acetonitrile (15%) in extractant, the mobile phase of water (0.1% formic acid)/acetonitrile, were optimized. The method was validated in terms of linearity (0.9990–0.9994), LOD (0.51 μg·kg−1), LOQ (1.73 μg·kg−1), intra-dayprecision (