Objective: To establish an analysis method for high performance liquid chromatography (HPLC) fingerprint of Aesculi Semen

detect the chemical constituents in Aesculi Semen from different origins and conduct cluster analysis

providing basis for its identification and quality control. Method: The conditions of separation and detection were achieved on an Agilent ZORBAX SB-C18 column(4.6 mm×250 mm

5 μm) with acetonitrile -0.2% phosphoric acid solution as the mobile phase for gradient elution. The flow rate was 1.0 mL·min-1; the detection wavelength was 220 nm; the column temperature was 30℃

and the injection volume was 10 μL. The similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A) was adopted for fingerprint analysis and similarity evaluation of the 36 batches of Aesculi Semen samples

and the identification method was discussed in combination with the characteristics of medicinal materials. SPSS 19.0 statistical software was used for cluster analysis. Result: A preferable method for fingerprint determination of Aesculus was established

with a similarity of 0.657-0.994.The medicinal materials were divided into four categories

including A. wilsonii

A. chinensis

A. chinensis var. chekiangensis and its adulterants A. wangii var. wangii. By comparing with control products

two chemical components were identified as escin A and escin B. 36 batches of these samples were also divided into 4 categories in cluster analysis

consistent with the similarity classification results. Conclusion: This study can be used for the determination of Aesculi Semen fingerprint and provides a theoretical basis for its further quality study.