A selective and sensitive UPLC-MS/MS method for determination of Galanthamine hydrobromide in the dog plasma was established.BEH C18 (50 mm×2.1 mm, 1.7 μm) was used as the solid phase, the mobile phase consisted of acetonitrile-0.01 mol·L-1 ammonium acetate (contained 0.1% formic acid) (40: 60).The flow rate was 0.1 mL·min-1, column temperature was 40°C, and inject volume was 3 μL.The monitoring ion of Galanthamine hydrobromide and internal standard clenbuterol hydrochloride was m/z 288.23→198.06, m/z 277.08→202.93, resp.A good linearity was shown in the range of 2.38-609.60 μg·L-1 (r=0.998, n=5).The average recovery was 102.3% with intra-day RSD and inter-day RSD less than 15%.Plasma matrix had no effect on quantification of Galanthamine hydrobromide.A simple, rapid, sensitive, selective and accurate UPLC-MS method was developed for the determination of galanthamine hydrobromide in plasma.