A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for the determination of vancomycin, norvancomycin and teicoplanin in milk samples. Selective reaction monitoring was performed and the external standard method was used for quantification. Among various solvents tested, 0.1% formic acid-acetonitrile (85:15, V/V) was selected. Samples were ultrasonically extracted and vancomycin and norvancomycin were cleaned up by cation exchange column chromatography, while teicoplanin was purified using a C18 solid-phase extraction (SPE) cartridge. Acetonitrile-0.1% formic acid solution was chosen as mobile phase for gradient elution. The analytes were separated on a reversed-phase C18 column before being analyzed by mass spectrometry. The limits of detection (LOD) for vancomycin, norvancomycin and teicoplanin were 2, 1 and 2 μg/kg, and the limits of quantitation (LOQ) were 4, 2 and 4 μg/kg, respectively. The recoveries of the method ranged from 77.3% to 84.5% at spiked levels equal to LOQ, 5 × LOQ and 10 × LOQ, with relative standard deviations (RSD) of 4.7%–7.2%. The method could be used in the qualitative and quantitative detection of vancomycin, norvancomycin and teicoplanin in milk samples.