Objective: To establish an HPLC chromatographic fingerprint analysis method for the quality evaluation of Calonyctii Semen. Method: The analysis was carried out by Agilent Extend-C18 column (4.6 mm×250 mm

5 μm)

with acetonitrile (A)-0.1% formic acid solution (B) as the mobile phase for gradient elution.The temperature of column was 30℃ at a flow rate of 1.0 mL · min-1; the detection wavelength was set at 300 nm

and the injection volume was 10 μL.The HPLC fingerprint was analyzed and the similarity was evaluated with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (version 2012).The comparison with reference drugs

cluster analysis

principal component analysis and partial least square discriminant analysis (PLS-DA) were used for analysis. Result: The precision

repeatability and stability of the established method can meet the requirements of fingerprint chromatography analysis.There were 14 common peaks in the HPLC fingerprint.The similarity of 10 batches of samples from Guangxi was greater than 0.70

indicating great differences in quality of Calonyctii Semen from different areas of Guangxi.Through the cluster analysis

principal component analysis and partial least square discriminant analysis (PLS-DA)

the samples were divided into three categories and showed that the producing origins and collection time were correlated with the quality of medicinal herbs. Conclusion: The developed HPLC fingerprint method is quick

simple

and could comprehensively reflect the chemical components information of alkaloids for Calonyctii Semen.Therefore

it could be used in quality evaluation of Calonyctii Semen.The samples collected in Yulin city and Guilin city have better quality

so these two cities can be the producing origins for Calonyctii Semen.