辣椒药材的HPLC指纹图谱建立及聚类分析和主成分分析

Establishment of HPLC Fingerprint,Cluster Analysis and Principal Component Analysis of Capsicum annuum

目的:建立辣椒药材的高效液相色谱(HPLC)指纹图谱,并进行聚类分析和主成分分析。方法:采用HPLC法。色谱柱为Agilent C18,流动相为甲醇-0.2%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,检测波长为235 nm,柱温为30℃,进样量为15μL。以辣椒素峰为参照,绘制15批药材样品的HPLC指纹图谱,采用《中药色谱指纹图谱相似度评价系统》(2004 A版)进行相似度评价,确定共有峰,并采用SPSS 19.0软件进行聚类分析和主成分分析。结果:15批药材样品的HPLC图谱相似度均在0.95以上;有12个共有峰,并指认了辣椒素峰;聚类分析结果显示,15批药材样品可聚为3类,S1、S3～S5、S7、S9～S13聚为一类,S2、S14、S15聚为一类,S6、S8聚为一类。经主成分分析,4个主成分因子的累积方差贡献率为94.093%,以S5药材样品的主成分因子综合得分最高、整体质量最好。结论:所建HPLC指纹图谱及聚类分析和主成分分析结果可为辣椒药材的质量控制提供参考。

OBJECTIVE:To establish HPLC fingerprint of Capsicum annuum,and to conduct cluster analysis and principal component analysis. METHODS:HPLC method was adopted. The determination was performed on Agilent C18 column with mobile phase consisted of methanol-0.2% phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 235 nm,and column temperature was 30 ℃. The sample size was 15 μL. Using capsaicin peak as reference, HPLC fingerprints of 15 batches of C. annuum from different production areas were determined. The similarity evaluation of common peaks was evaluated by using the TCM Chromatographic Fingerprint Similarity Evaluation System(2004 A edition)to confirm common peaks. Cluster analysis and principal component analysis were performed by using SPSS 19.0 software. RESULTS:The similarities were more than 0.95 in HPLC chromatograms of 15 batches of C. annuum. There were 12 common peaks. Its HPLC fingerprint was in good agreement with that of control fingerprint. 15 batches can be divided into three sub-categories as S1,S3-S5,S7,S9-S13 sub-categorie,S2,S14,S15 sub-categorie and S6,S8 sub-categorie. Through the principal component analysis,the cumulative contribution rate of 4 main component factors was 94.093,and comprehensive score of S5 was the highest with the best quality. CONCLUSIONS:Established HPLC fingerprints,cluster analysis and principal component analysis results can provide reference for the quality control of C. annuum.