Abstract: An analytical method was developed for determination of 7 aminoglycosides antibiotics in bear bile powder by hydrophilic interaction liquid chromatography tandem mass spectrometry. The samples were purified by mix-mode weak cation exchange and reversed-phase SPE. Waters ACQUITY UPLC BEH Amide column (100 mm × 3.0 mm, 1.7 μm) was used with 0.2% formic acid aqueous solution-0.2% formic acid acetonitrile solution as mobile phases by gradient elution. The aminoglycosides were detected by electrospray ionization mass spectrometry in positive mode with multiple reaction monitoring (MRM) mode. Spectinomycin, streptomycin, amikacin, kanamycin, tobramycin, apramycin and neomycin possessed good linear correlation in the respective concentration ranges, with the correlation coefficients more than 0.99. The mean recoveries at 3 spiked levels were in the range of 61.3%~127.3%, and the RSDs were 0.1%~1.9%. The limits of quantification were 0.2~1.0 mg·kg^-1. The method had been applied to the analysis of actual samples.