An HPLC method for determination of flavonol glycosides in Shuxuening for injection was presented.Dikma Diamonsil C18 (200 mm × 4.6 mm, 5 μm) column was used as chromatog. column, and methanol-0.5% acid water (54:46) as mobile phase.The detection wavelength was set at 360 nm, flow rate at 1.0 mL/min-1, and column temperature at 25°.There was a good linear relationship when Quercetin of Total Favonol Glycosides was in the range of 0.007 1-0.085 2 mg/mL-1 (r = 0.999 0, n = 5), Kaempferol in the range of 0.006 5-0.078 0 mg/mL-1 (r = 0.999 7, n = 5), and Isorhamnetin in the range of 0.001 5-0.018 0 mg/mL-1 (r = 0.998 1, n = 5).The average recoveries of Total Favonol Glycosides, Quercetin, Kaempferol and Isorhamnetin were resp. 101.2% (RSD = 0.5%), 99.2% (RSD = 0.9%), 103.8% (RSD = 0.8%), and 102.7% (RSD = 2.0%).This method is accurate in determination results, simple in operation, high in sensitivity and good in reproducibility, and can be used for the quality control of the preparation