To establish a method to determine the content of Zhikang Granules with the naphthopyranone reference extract(NRE) and reference substance in Cassiae Semen and compare and analyze the results to explore the applicability and feasibility of the application of Chinese medicinal reference extract instead of monomer reference substance in the quality control of Chinese medicinal compound.Methods:Four reference substances,rubrofusarin-6-O-β-D-gentiobioside,cassiaside B2,cassiaside C,and cassiaside C2,and NRE with known content were used as the controls,and the content of the four components in Zhikang Granules was determined by HPLC.The results obtained by the two methods were analyzed by the t-test.Results:The experimental results showed that the content of the four components were significantly different between the reference extract method and the reference substance method(P>0.05).Conclusion:NRE can be used as the reference substance in the quality control of Zhikang Granules.The findings of this study are expected to provide a scientific basis for the application of NRE in the quality evaluation of Chinese medicinal compounds containing Cassiae Semen,and verify the feasibility and applicability of Chinese medicinal reference extracts instead of chemical controls used in the quality control of Chinese medicinal compounds.