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To control the quality of Euonymus fortunei (Turcz.) Hand.-Mazz., a simple and reliable method of high-performance liquid chromatography (HPLC) coupled with photodiode array detector (PAD) was developed for both fingerprint analysis and quantitative determination. Four representative flavonoids, namely, kaempferol-3-O-β-D-glucopyranosyl-(1→4)-α-L-rhamnopyranosyl-7-O-β-D-glucopyranosyl-(1→4)-α-L-rhamnopyranoside (I), kaempferol-3,7-O-α-dirhamnopyranoside (II), apigenin-7-O-β-D-glucopyranoside (III), and kaempferol-3-(4″-O-acetyl)-O-α-L-rhamnopyranoside-7-O-α-L-r hamnopyranoside (IV) isolated from E. fortunei, were used as reference compounds and simultaneously determined by the validated HPLC method. The unique properties of the chromatographic fingerprint were validated by analyzing 11 batches of E. fortunei, E. japonicus, E. laxiflorus, E. myrianthus, and E. hamiltonianus samples. Our results revealed that the chromatographic fingerprint combined with similarity measurement could efficiently identify and distinguish E. fortunei from the other investigated Euonymus species.

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