Abstract: A method for simultaneous determination of multiple antibiotic residues in honey was developed by means of a series mass spectrometry(MS)methods.After the sample was extracted from 1%acetonitrile formate-Na2EDTA aqueous solution,the containing salt was removed from the water,and the C18 adsorbent was added to the upper liquid to purify it.After concentrated and dissolved,the solution was filtered by a membrane filter.The prepared samples were separated by the C18 chromatographic column(2.1 mm×100 mm,1.7μm),and 0.1%formic acid aqueous solution and methanol solution were used as the flow phase for gradient elution.The UPLC-MS/MS method was used to qualitatively and quantitatively determine multiple antibiotic residues in honey simultaneously.The results showed that the linear relationship of the drugs to be tested was good in the range of 0.01μg/mL to 5.00μg/mL,and the correlation coefficient(r)was all above 0.99.Addition recovery experiments were conducted at 1,5 and 10μg/kg concentration levels,the average recovery rate was between 68.0%and 104.0%,and the relative standard deviation ranged from 3.2% to 15.5%.The detection limit of the developed method reached up to 1μg/kg.The method has good reproducibility,high sensitivity,short analysis time and strong corroboration ability,which is recommended to be used in simultaneous determination of multiple antibiotics residues in honey.