杜仲补天素丸的HPLC指纹图谱建立、化学模式识别分析及含量测定

目的:建立杜仲补天素丸的指纹图谱并进行化学模式识别分析,测定杜仲补天素丸中7个成分的含量,为该制剂的质量控制提供参考.方法:采用高效液相色谱法(HPLC),以Pntulips BP-C18 Plus为色谱柱,以0.2％磷酸水溶液-乙腈为流动相进行梯度洗脱,流速为1.0 mL/min,检测波长为330 nm,柱温为35℃...

Establishment of HPLC Fingerprint,Chemical Pattern Recognition Analysis and Content Determination of Duzhong Butiansu Pill

OBJECTIVE:To establish fingerprint of Duzhong butiansu pills,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-C18 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 20μL.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 S11 were classified as one category,S2,S10 and S12 were clustered into one category,S6 was one category and S8 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 contents of above 7 components were 18.1962-31.9513,0.0006-0.0494,0.2348-0.4159,0.0395-0.0791,0.0535-0.2493,0.0005-0.0008,0.6464-1.1469 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.