High-performance liquid chromatography is employed to determine the contents of five mark components, safflor yellow A, puerarin, ferulic acid, ginsenoside Rg1, and Rb1, in the Traditional Chinese Medicinal preparation Naodesheng injection. The separation is performed on a C18 column by stepwise gradient elution with water (0.1%, v/v, phosphoric acid)-acetonitrile (0 min, 86:14; 48 min, 75:25; and 68 min, 50:50) as the mobile phase at a flow rate of 1.0 mL/min, with UV detection at 203 nm. Five regression equations show a good linear relationship between the peak area of each marker and concentration. The recoveries of the markers listed are 99.6%, 100.2%, 99.7%, 100.0%, and 99.7%, respectively. The repeatability and reproducibility (relative standard deviation) of the method are less than 1.4% and 1.8%, respectively.