Abstract: Luminol solution of 2.0×10^-5mol·L^-1 was oxidized by potassium permanganate solution of 1.5×10^-4mol·L^-1 in strong alkaline condition to make chemiluminescence reaction, which could be sensitized by rutin, and the quantification of rutin was carried out by detecting its chemiluminescence intensity. Based on this fact, a method for the determination of rutin by high performance liquid chromatography coupled with chemiluminescence detection was established. Rutin tablets were grinded into powder and dissolved with methanol. The sample solution was prepared by reflux heating, 0.45 μm filter membrane filtration and ultrasonic oscillation. BPCL weak luminescence measuring instrument was modified, and coupled with high performance liquid chromatography and chemiluminescence detection system. In the chemiluminescence condition, the flow rate was adjusted to 4 cm·s^-1, and the length of the detected coil was 10 cm. Using C₁₈ column as stationary phase and a mixture of methanol and 2% (φ) glacial acetic acid solution at volume ratio of 1∶1 as mobile phase, rutin could be separated well. The results showed that linear relationship between mass concentration of rutin and its chemiluminescence intensity was found in the range of 1.00-150.00 μg·L^-1, with detection limit of 1.00×10^-6g·L^-1. RSDs (n=11) of the determined values was 2.7%.