An ultra performance liquid chromatography-tandem mass spectrometric(UPLC-MS/MS) method was developed for the simultaneous determination of spiramycin(e.g.spiramycin Ⅰ，SPM-Ⅰ) and neospiramycin(e.g.neospiramycin Ⅰ，NSPM-Ⅰ) residues in antibiotic production wastewater.The factors influencing the recoveries of antibiotics，including organic extractant type，water sample pH value，extraction time and ratio of water sample to organic extractant，were investigated.The target compounds were separated on an ACQUITY UPLC BEH C18 column(100 mm × 2.1 mm i.d.，1.7 μm) by gradient elution using 0.1% formic acid(A) and acetonitrile(B) as mobile phases at a flow rate of 0.2 mL/min.The qualification and quantification of target antibiotics were carried out by MS/MS with positive electrospray ionization under multiple reaction monitoring(MRM) mode.The calibration curves were linear over an antibiotic concentration range of 0.02-1.00 mg/L with correlation coefficients(r2) more than 0.999.The recoveries of antibiotics ranged from 70% to 88% with relative standard deviations(RSD，n=6) less than 10%.The detection limits(S/N=3) for SPM-Ⅰ and NSPM-Ⅰ were 0.03 μg/L and 0.06 μg/L，respectively.The developed method was applied in the determination of antibiotic production wastewater from a pharmaceutical manufacturing factory in Wuxi City，with SPM-Ⅰ of (115.6±2.7) mg/L and NSPM-Ⅰ of(5.3±0.4) mg/L.