A method for the determination of both norepinephrine bitartrate injection and phenylephrine hydrochloride injection and their related substances was established.A HPLC was performed on a column of Kromasil C18 (250 mm×4.6 mm, 5 μm) with the mobile phase of methanol-acetonitrile-sodium heptane solution (1: 1: 8) at detection wavelength of 280 nm, flow rate of 1.0 mL/min and column temperature of 30°C.For norepinephrine bitartrate, the calibration curve was linear (r=0.999 8) over the range of 80-320 μg/mL, the average recovery was 101.2% with RSD 0.18% and the limit of detection was 152.3 ng/mL.For phenylephrine hydrochloride, the calibration curve was linear (r=0.999 6) over the range of 100-400 μg/mL, the average recovery was 98.1% with RSD 0.051% and the limit of detection was 103.8 ng/mL.This method is simple, accurate, sensitive and good reproducible, which can be applied in the determination of both norepinephrine bitartrate injection and phenylephrine hydrochloride injection and their related substances.