Abstract:This study developed an ultra-high performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） method based on multiple secondary interactions of partition， which was dominated， dipole interaction， hydrogen bonding， adsorption， and ion interaction for retention and separation to detect the residues of 16 aminoglycoside veterinary drugs in pork. The samples were extracted by phosphate buffer solution（pH4.0） containing 2% trichloroacetic acid， 0.04 mmoL/L ethylenediaminetetraacetic acid disodium salt（EDTA-2Na）， 0.05 moL/L sodium pentanesulfonate， and 0.05 moL/L sodium hexanesulfonate， purified by HLB solid-phase extraction column （6 mL/200 mg）， retained and separated by an Agilent Hilic plus column（2.1 mm × 100 mm， 3.5 μm） with the use of hydrophilic interaction， and detected by a triple quadrupole tandem mass spectrometer. There was a good linear correlation in the range of 50-1 500 ng/mL with correlation coefficients ≥ 0.993. The limit of detection was 3.8-19.7 μg/kg， and the limit of quantitation was 12.5-65.8 μg/kg.At the spiking levels of 50， 250， 500 μg/kg， the average recovery was 79.6%-115.3%， and the relative standard deviation （RSD） was 0.8%-14.6%. This method can simultaneously detect 16 aminoglycoside antibiotics in pork with simple pretreatment， good stability， high accuracy， and high sensitivity， which can meet the regulatory requirements for the detection of aminoglycoside residues in pork.