High-performance liquid chromatography with ultraviolet spectrometer (HPLC-UV) was used to simultaneously detect the four flavone C-glycosides, i.e. orientin, isoorientin, vitexin and isovitexin. Analytes were separated with Waters XTerra MS C18 column (250 mm 4.6 mm, 5 μm) using acetonitrile and 0.5% (φ) formic acid as mobile phase. The flow rate was set at 1.0 mL·min-1 with the column temperature at 30 ℃, and the detection wavelength was 360 nm. The calibration curve was linear over the concentration range of 0.1~10.0 mg·L-1 for the mixed standard solution. Analytes were separated in 22 minutes, and the relative standard deviation values were all above 0.999. LOD values of standards were found to be between 0.03 and 0.07 mg·L-1, and LOQ values were in the range of 0.04~0.08 mg·L-1. After comparing the spectra (240~400 nm) of four flavone C-glycosides in mixed standards and the final product purified by macroporous resin, respectively, the curve shape and characteristic ultraviolet absorption wavelength of each flavone C-glycoside including orientin, isoorientin, vitexin and isovitexin were fitted well. The bamboo leaves sample was extracted by ethanol under reflux, and then partitioned with water and petroleum ether. The aqueous phase was added onto macroporous resin(AB-8), and the fraction of ethanol-water (40%, φ) was concentrated. It was found that the contents of orientin, isoorientin, vitexin and isovitexin relative to the fraction of ethanol-water were 13.73, 49.68, 7.85 and 30.70 mg·g-1, respectively. In addition, the average recovery of the four flavone C-glycosides ranged from 34.90% to 87.64% with RSD values from 0.41% to 10.83%. The results showed that bamboo leaves sample had good stability and repeatability. The new method was used to analyze the four flavone C-glycosides quickly and provide quality control for commercial products.