摘要
高效液相色谱法(HPLC)测定对不同来源五倍子两种形态肚倍和角倍进行HPLC特征指纹图谱研究,并通过聚类分析和主成分分析深入研究五倍子药材不同形态中有效成分差异,为五倍子药材质量控制做奠基.采用HPLC梯度洗脱法对肚倍和角倍样品进行测定,并记录色谱图,按《中药色谱指纹图谱相似度评价系统(2012 版)》对色谱图进行匹配生成肚倍和角倍的特征图谱,进行相似度分析.采用化学计量学软件origin 2018 对肚倍和角倍进行聚类分析、主成分分析(PCA)研究.结果显示:分别建立了肚倍和角倍的 HPLC 特征指纹图谱共有模式,标识了没食子酸、没食子酸甲酯、1,2,3,6-四-O-没食子酰葡萄糖、1,2,3,4,6-五-O-没食子酰葡萄糖4 个特征峰.角倍与肚倍相似度低,可通过聚类分析、主成分分析将其分为 2 大类.从两者的HPLC特征指纹图谱相比,肚倍比角倍多了5 个峰,保留时间为 20.3、26.0、27.6、54.6、75·9 min,同时少了 49.1 min相对位置的保留峰.通过分析不同产地的五倍子角倍和肚倍中特征性化学成分,表明肚倍的有效成分多于角倍.利用相似度研究、聚类分析技术及主成分分析,可有效区分二者,为五倍子药材质量综合评价和利用提供参考.
Abstract
To use high-performance liquid chromatography(HPLC)to determine the HPLC characteristic fingerprints of two different forms of Galla chinensisfrom different producing areas,and to study the differences in effective components in different forms of Chinese gallnuts through cluster analysis and principal component analysis,lay the foundation for the quality control of gallnut.HPLC gradient elution method was used to determine the samples of horned gallnut and belly gallnut,and the chromatograms were recorded.The chromatograms were matched according to the " Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints(2012 Edition)" to generate characteristic spectra of horned gallnut and belly gallnut,and similarity analysis was performed.Cluster analysis and principal component analysis(PCA)were conducted on the abdomen and horn polyploids using the chemometric software Origin 2018.The results showed that a common pattern of HPLC characteristic fingerprints for horned gallnut and belly gallnut was established,identifying four characteristic peaks:gallic acid,methyl gallate,1,2,3,6-tetra-O-glycerol,and 1,2,3,4,6-penta-O-glycerol.The similarity between Jiaobei and Beibei is low,and they can be divided into two categories through cluster analysis and principal component analysis.Compared with the HPLC fingerprint of the two,belly gallnut has 5 more peaks,with retention times of 20.3 minutes,26.0 minutes,27.6 minutes,54.6 minutes,and 75.9 minutes,and a retention peak of 49.1 minutes in relative position is missing.By analyzing the characteristic chemical components in Chinese gallnut and Tripterygium from different regions,it is shown that belly gallnut has more effective components than horned gallnut.By using similarity research,cluster analysis techniques,and principal component analysis,the two can be effectively distinguished,providing reference for the comprehensive evaluation and utilization of the quality of Chinese gallnut medicinal materials.
基金项目
中央级公益性科研院所基本科研业务费专项(Y2022YJ14)
兰州市科技计划(2022-2-61)
湖北省揭榜制科技项目(2022BEC033)
兽用天然药物与抗生素替代创新工程项目(25-LZIHPS-03)