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AbstractCalyx of Diospyros kaki Thunb. (DKC), a famous traditional Chinese medicine, has a remarkable therapeutic effect on hiccups but its quality control is insufficient. In the study, a method was established by ultra-high performance liquid chromatography – mass spectrometry (UHPLC-MS). 17 peaks were observed in the mass spectrum and their chemical structures were preliminarily identified, including 3 organic acids and 14 flavonoids. The reference UHPLC fingerprint was built based on 25 batches of DKC samples, containing 15 common peaks. 11 batches of DKC samples from Guangxi and Hebei were superior to other 14 batches, sharing high similarity from 0.966 to 0.988 with the reference fingerprint. The chemometric methods, including hierarchical clustering analysis (HCA), principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA), were used to distinguish the DKC samples. The recognition methods clearly distinguished the medicinal materials from different regions, indicating that the main chemical constituents leading to the differences were protocatechuic acid, 4-hydroxybenzoic acid, rutin (presumptive result based on MS data), isoquercitrin, kaempferol, and hyperoside. The established fingerprint method is simple, rapid, repeatable, and specific and provides a reference for the quality evaluation of DKC. [ABSTRACT FROM AUTHOR]