Objective: To establish a method for the determination of Panax notoginseng saponins and icariin in Guyu Injection.Method: Diamonsil C18 column(4.6 mm × 250 mm,3.5μm) was used with acetonitrile and water as mobile phase with gradient elution;the flow rate was 1 mL.min-1;detection wavelength was set at 203 nm;the column temperature was maintained at 25 ℃.Result: The linear range of icariin was within 1.0610.60 μg and the average recovery of assay was 100.2%(RSD 1.54%);the linear range of notoginsenoside R1 was within 0.42-4.20 μg and the average recovery of assay was 100.6%(RSD 1.89%);the linear range of ginsenoside Rg1 was within 1.90-18.97 μg and the average recovery of assay was 99.9%(RSD 1.09%);the linear range of ginsenoside Re was within 0.18-1.78 μg and the average recovery of assay was 100.8%(RSD 1.96%);the linear range of ginsenoside Rb1 was within 2.23-22.34 μg and the average recovery of assay was 101.4%(RSD 1.83%);the linear range of ginsenoside Rd was within 0.45-4.53 μg and the average recovery of assay was 101.2%(RSD 2.37%).Conclusion: The method is accurate and reproducible.It can be used for determination of Panax notoginseng saponins and icariin in Guyu Injection.