The invention provides a construction method of a rheum lhasaense fingerprint spectrum and a standard fingerprint spectrum of rheum lhasaense. High performance liquid chromatograph is conducted on a rheum lhasaense sample through a chromatographic column taking octadecylsilane chemically bonded silica as packing, the fingerprint spectrum is constructed by taking piceatannol 3'-O-glucoside as a reference peak, and the fingerprint spectrum is evaluated on this basis. The obtained rheum lhasaense fingerprint spectrum can comprehensively, systematically and characteristically reflect composition ingredients in rheum lhasaense, comprehensively and effectively evaluate the quality of rheum lhasaense and objectively reflect authenticity and merits of a product and is beneficial for product quality monitoring. The method is easy and convenient to operate, stable, high in precision and good in reproducibility.