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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
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篇名: 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
TITLE: Establishment of HPLC Fingerprint ,Chemical Pattern Recognition Analysis and Content Determination of Duzhong Butiansu Pill
摘要: 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PntulipsBP-C18Plus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1.0mL/min,检测波长为330nm,柱温为35℃,进样量为20μL。以丹皮酚为参照,使用《中药色谱指纹图谱相似度评价系统App(2012版)》建立12批杜仲补天素丸(S1~S12)的指纹图谱,确定共有峰并进行相似度评价,通过与对照品比对进行色谱峰指认。采用SPSS21.0App和SIMCA13.0App进行聚类分析和主成分分析,对22个共有峰进行评价。采用上述HPLC法测定12批样品中指认成分的含量。结果:从12批杜仲补天素丸的HPLC指纹图谱中标定共有峰22个,相似度均不低于0.960;共指认化学成分7个,分别为没食子酸(峰1)、绿原酸(峰3)、甘草苷(峰6)、金丝桃苷(峰7)、毛蕊花糖苷(峰8)、淫羊藿苷(峰14)和丹皮酚(峰15)。12批样品中,S1、S3~S5、S7、S9、S11聚为一类,S2、S10、S12聚为一类,S6聚为一类,S8聚为一类;22个共有峰被分为3个主成分,主成分1的特征值(15.130)和方差贡献率(68.775%)最大,其中峰3(0.305)和峰4(0.298)对应成分的得分系数最高。12批样品中,上述7个成分的含量分别为18.1962~31.9513、0.0006~0.0494、0.2348~0.4159、0.0395~0.0791、0.0535~0.2493、0.0005~0.0008、0.6464~1.1469mg/g。结论:成功建立了杜仲补天素丸的HPLC指纹图谱。12批样品被聚类分成4类,峰3(绿原酸)和峰4(未知)可能是造成样品差异的重要因素。7个成分中没食子酸的含量最高。
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.
期刊: 2021年第32卷第08期
编辑: 刘敏,曹国琼,张仕林,葛秋平,刘娅,张永萍
AUTHORS: LIU Min,CAO Guoqiong ,ZHANG Shilin,GE Qiuping ,LIU Ya,ZHANG Yongping
关键字: 杜仲补天素丸;高效液相色谱法;指纹图谱;化学模式识别;含量测定
KEYWORDS: Duzhong butiansu pills ;HPLC;Fingerprint;Chemical pattern recognition ;Content determination
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