Establishment of HPLC Fingerprint ，Chemical Pattern Recognition Analysis and Content Determination of Duzhong Butiansu Pill / 中国药房

ABSTRACT

OBJECTIVE：To estab lish fingerprint of Duzhong butiansu pill s，analyze its chemical pattern recognition ，and determine the contents of 7 components in Duzhong butiansu pills ，so as to provide reference for the quality control of the preparation. METHODS ：HPLC method was adopted. The determination was performed on Pntulips BP-C 18 Plus column with 0.2％ phosphoric acid water-acetonitrile as mobile phase （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 330 nm，and column temperature was 35 ℃. The sample size was 20 μL. With paeonol as the reference，the HPLC fingerprints of 12 batches of Duzhong butiansu pills （S1-S12） were established with Similarity Evaluation System for TCM Chromatographic Fingerprint （2012 edition）； common peaks were determined and the similarity was evaluated. The chromatographic peaks were identified by comparing with the reference substance. SPSS 21.0 and SIMCA 13.0 software were used for cluster analysis and principal component analysis ，and 22 common peaks were evaluated. The contents of the identified components in 12 batches of samples was determined by the above HPLC method. RESULTS ：A total of 22 common peaks were identified in the HPLC fingerprint of 12 batches of Duzhong butiansu pills ，and the similarity was no loss than 0.960. There were 7 chemical components identified ，which were gallic acid （peak 1），chlorogenic acid （peak 3），liquiritoside（peak 6），hyperoside （peak 7），verbascoside（peak 8），icariin（peak 14）and paeonol （peak 15）. Among the 12 batches of samples ，S1，S3-S5，S7， S9 and S 11 were classified as one category ，S2，S10 and S 124Y091 were clustered into one category ，S6 was one category and S was one category. The 22 common peaks were divided into three principal components. The characteristic value （15.130） and contribution rate （68.775％） of principal component 1 were the largest ，and the score coefficients of peak 3（0.305）and peak 4（0.298）were the highest. Among 12 batches of samples，the cont ents of above 7 components were 18.196 231.951 3，0.000 6-0.049 4，0.234 8-0.415 9，0.039 5-0.079 1，0.053 5-0.249 3，0.000 5-0.000 8，0.646 4-1.146 9 mg/g，respectively. CONCLUSIONS：HPLC fingerprint of Duzhong butiansu pills is established successfully. Twelve batches of samples are clustered into 4 category. Peak 3（chlorogenic acid ）and peak 4（unknown）may be the important factors causing the difference of samples. The content of gallic acid is the highest among the 7 components.