Objective The aim of this study was to construct high performance liquid chromatography (HPLC) fingerprint of Seabuckthorn granules, and quantitatively analyze the key active components by one test and multiple evaluation (QAMS) technique, so as to achieve systematic evaluation of seabuckthorn granules quality.Methods The fingerprints of Seabuckthorn granules were established by HPLC, and analyzed by similarity evaluation and principal component analysis (PCA). Using quercetin as internal reference, the QAMS model of kaempferol and isorhamnetin content was constructed, and the results were compared with those of external standard method (ESM).Results There were 13 common peaks in the HPLC fingerprints of 9 batches of Seabuckthorn granules, and quercetin, kaempferol and isorhamnetin were identified with similarity ranging from 0.92 to 1.0. PCA analysis showed that the main differences of sea buckthorn granules were peak 1(quercetin), peak 2(kaempferol) and peak 3(isorhamnetin). In addition, there was no significant difference between QAMS method and ESM method in the analysis results of the three components.Conclusion The HPLC fingerprint and QAMS method established in this study are simple, efficient, accurate and durable, and provide a reliable reference for the quality evaluation of Seabuckthorn granules.