Abstract: [Objective]To establish a HPLC method for determination of lignan in flaxseed. [Methods]The samples were treated with liquid-liquid extraction; the standard substance of lignan was used to conduct quantitative analysis; Agilent Cls Sym chromatographic column （250 mm×4.6 mm） was used to isolate; aqueous solution of methanol （methanol：water=4：6） was used as mobile phase; lignan were detected by UV detector with wavelength of 290 nm; methodology validation were conducted by using standard addition recovery test and precision test. [Results]The recovery of lignan in flaxseed ranged from 95% to 108%, and the RSD of precision was 7.43%. The rechecked results of determination of lignan in flaxseed by the established HPLC method from different laboratories were similar, which proved the good repeatability of the method. [ Conclusion ]The established HPLC method could be used for determination of lignan in flaxseed and provided reliable experimental basis for the industrial lignan purification.