Abstract: Objective : A reversed- phase high performance liquid chromatographic method ( RP- HPLC) was developed for determination of different configuration of vitamin E in Euryale Semen. Methods:RP-HPLC determination was performed on a Waters Xbridge C18column (4.6mm×150mm, 5μm) , Methanol-water (97∶3) was used as the mobile phase at the flow rate of 1.00mL/min, the column temperature was kept as 30℃ and the detection wavelength was set as 295nm. Results : α-tocopherol, β +γ-tocopherol, δ-tocopherol were separated in baseline, the linear range was 0.12¹.2g/L for α-tocopherol, 0.056⁰.56g/L for β+γ-tocopherol and 0.0096⁰.096g/L for δ-tocopherol. R2were 0.9998, 0.9999, 0.9998, the average recoveries were 100.47%, 100.04%, 99.10%. RSD were 0.27%, 4.65%, 1.92%. The total Vitamin E content of the sample of Anhui was the highest with 2.2633mg/g. Conclusion:This method was sample, accurate and with good reproducibility, which could be used as Euryale Semen quality control of medicine and also provides a scientific reference for the rational use of Euryale Semen.