基于高效液相色谱和红外光谱的生三七及其炮制品的鉴别

Identification of Panax notoginseng and Its Processed Products Based on HPLC and IR Spectrum

目的:鉴别生三七及其炮制品。方法:采用高效液相色谱法(HPLC)建立指纹图谱。以人参皂苷Rb1为参照,采用《中药色谱指纹图谱相似度评价系统(2012版)》绘制15批生三七及其炮制品的HPLC指纹图谱并进行相似度评价,通过与对照品对比确定共有峰。采用SPSS 21.0、SIMCA 14.1软件进行聚类分析、主成分分析、正交偏最小二乘法分析,以变量重要性投影(VIP)值大于1为标准,筛选造成生三七及其炮制品质量差异的标志性成分。采用OMNIC 8.2.0软件建立生三七及其炮制品的红外(IR)指纹图谱并进行相似度评价,采用双指标序列分析法对15批生三七及其炮制品的IR指纹图谱吸收峰进行分析。结果:15批生三七有16个共有峰,相似度为0.911~1.000;炮制品有25个共有峰,相似度为0.862~1.000;二者共有12个相同的共有峰,并指认其中3个共有峰,分别为三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1。聚类分析结果显示,当距离为10时,15批生三七可聚为两类,SW1~SW5聚为一类,SH1~SH5、SQ1~SQ5聚为一类;15批炮制品ZW1~ZW5、ZH1~ZH5、ZQ1~ZQ5聚为一类。当距离为5时,15批生三七可聚为3类,SW1~SW5聚为一类,SH2~SH5、SQ2聚为一类,SQ1、SQ3~SQ5、SH1聚为一类;15批炮制品可聚为两类,ZW1~ZW5聚为一类,ZH1~ZH5、ZQ1~ZQ5聚为一类。主成分分析结果显示,前两个主成分的累计方差贡献率为80.104%。正交偏最小二乘法分析结果显示,5个峰的VIP值大于1,分别为峰H、峰G、峰J、峰F(人参皂苷Rg1)、峰I。15批生三七及其炮制品IR指纹图谱的相似度分别为0.8897~1.0000、0.9728~1.0000;共有峰率均为80%~100%,变异峰率分别为0~17.65%、0~18.75%。经吸收峰波数对比发现,生三七在3440、1450 cm^(-1)处,炮制品在1530、575 cm-1处存在差异。结论:所建HPLC指纹图谱和IR指纹图谱的相似度均较好,能有效区分生三七及其炮制品。不同产地生三七及其炮制品的共有峰率高、变异率低,二者化学成分不同;峰H、峰G、峰J、人参皂苷Rg1、峰I为引起生三七及其炮制品质量差异的标志性成分。

OBJECTIVE:To identify Panax notoginseng and its processed products.METHODS:The fingerprint was established by HPLC.Using ginsenoside Rb1 as reference,HPLC fingerprints of 15 batches of P.notoginseng and its processed products were drawn and the similarity evaluation was conducted by using the Similarity Evaluation System for TCM Chromatographic Fingerprints(2012 edition).The common peaks were confirmed by comparing with substance control.SPSS 21.0 and SIMCA 14.1 software were used to perform cluster analysis,principal component analysis and orthogonal partial least squares-discriminant analysis;taking the variable importance projection(VIP)value greater than 1 as the standard,the differential marker components causing the quality difference between P.notoginseng and its processed products were screened.IR fingerprints of P.notoginseng and its processed products were established by OMNIC 8.2.0 software,and the spectral similarity was evaluated;double index sequence analysis was used to analyze absorption peaks of IR fingerprints of 15 batches of P.notoginseng and its processed products.RESULTS:There were 16 common peaks in the fingerprints of 15 batches of P.notoginseng,and the similarities were 0.911-1.000;there were 25 common peaks in the fingerprints of processed products,and the similarities were 0.862-1.000.They had 12 identical common peaks,and three of them were identified as sanchinoside R1,ginsenoside Rg1 and ginsenoside Rb1.Results of cluster analysis showed that when the distance was 10,15 batches of P.notoginseng could be clustered into two categories,SW1-SW5 into one category,SH1-SH5 and SQ1-SQ5 into one category,ZW1-ZW5,ZH1-ZH5 and ZQ1-ZQ5 of 15 batches of processed products could be clustered into one category.When the distance was 5,15 batches of P.notoginseng could be clustered into three categories,SW1-SW5 into one category,SH2-SH5 and SQ2 into one category,SQ1,SQ3-SQ5 and SH1 into one category.Fifteen batches of processed products could be clustered into two categories,ZW1-ZW5 into one category,ZH1-ZH5 and ZQ1-ZQ5 into one category.The results of principal component analysis showed that the cumulative variance contribution rate of the first two principal components was 80.104%.The results of orthogonal partial least squares-discriminant analysis showed that the VIP values of the five peaks were greater than 1,which were peak H,peak G,peak J,peak F(ginsenoside Rg1)and peak I.The similarity of IR fingerprints of 15 batches of P.notoginseng and its processed products were 0.8897-1.0000 and 0.9728-1.0000;the common peak rates were 80%-100%,and the variation peak rates were 0-17.65% and 0-18.75%,respectively.By comparing the wave numbers of absorption peaks,it was found that there were differences between P.notoginseng at 3440 and 1450 cm^(-1) and processed products at 1530 and 575 cm-1.CONCLUSIONS:Established HPLC fingerprint and IR fingerprint have good similarity,and could effectively distinguish P.notoginseng and its processed products.P.notoginseng and its processed products from different habitats have high common peak rate and low variation rate,and their chemical components are different;peak H,peak G,peak J,ginsenoside Rg1 and peak I are differential marker components causing the quality difference between P.notoginseng and processed products.