不同产地显齿蛇葡萄以及近缘种指纹图谱的建立及有效成分分析  

HPLC fingerprint and chemical pattern recognition of Ampelopsis grossedentatafrom different producing areas

作　　者：凌伟红 陈燕 冯淼 王超纯 龚慧敏 田春莲[1,3,4] LING Wei-hong;CHEN Yan;FENG Miao;WANG Chao-chun;GONG Hui-min;TIAN Chun-lian(Hunan Key Laboratory of Forest Products and Chemical Industry Engineering,Jishou University,Zhangjiajie 427000,China;Zhangjiajie Yongding County Agricultural and Rural Bureau,Zhangjiajie 427000,China;College of Biology and Environmental Sciences,Jishou University,Jishou 416000,China;National and Local United Engineering Laboratory of Integrative Utilization Technology of Eucommia ulmoides,Jishou University,Jishou 416000,China)

机构地区：[1]吉首大学林产化工工程湖南省重点实验室,湖南张家界427000 [2]张家界市永定区农业农村局,湖南张家界427000 [3]吉首大学生物资源与环境科学学院,湖南吉首416000 [4]杜仲综合利用技术国家地方联合工程实验室,湖南吉首416000

出　　处：《中草药》2023年第17期5725-5733,共9页Chinese Traditional and Herbal Drugs

基　　金：湖南省科技厅资助项目(2019NK4164);林产化工工程湖南省重点实验室开放基金(LCHG21908)。

摘　　要：目的建立显齿蛇葡萄Ampelopsis grossedentata以及近缘种的高效液相色谱(HPLC)指纹图谱,并对有效成分进行定量分析。方法使用Ultimate XB-C18(250 mm×4.6 mm,5μm)色谱柱;流动相为乙腈-0.01%磷酸,检测波长为280、360 nm,体积流量为1 mL/min,进样量20μL,柱温30℃下梯度洗脱,建立显齿蛇葡萄以及近缘种HPLC指纹图谱,结合相似度评价、层次聚类分析(clustering analysis,CA)、主成分分析(principal component analysis,PCA)与正交偏最小二乘法-判别分析(orthogonal partial least squares method-discriminant analysis,OPLS-DA)等化学模式识别方法,评价不同产地显齿蛇葡萄及近缘种的质量。结果建立的HPLC指纹图谱方法符合方法学要求,18批显齿蛇葡萄属样品指纹图谱有31个共有峰,经对照品定性指认4、9、11、14、15和24号峰对应的化合物分别为没食子酸、儿茶素、二氢杨梅素、芦丁、杨梅苷和杨梅素,且相似度>0.8;所有样品按产地经CA分为2类,PCA结果与其一致;OPLS-DA筛选出不同产地显齿蛇葡萄及近缘种的13~14个差异标志物,以二氢杨梅素和杨梅素为主要差异标志物。结论建立的指纹图谱方法稳定、可靠、重复性好,结合化学模式识别可用于作为藤茶原料的蛇葡萄属植物的质量评价。Objective To establish the HPLC fingerprint of Xianchi Sheputao(Ampelopsis grossedentata)and perform chemical pattern recognition.Methods The gradient elution was carried out on an Ultimate XB-C18 column(250 mm×4.6 mm,5μm)with a mobile phase of acetonitrile-0.01%phosphoric acid,detection at 280 nm and 360 nm,a volume flow rate of 1 mL/min,an injection volume of 20μL and a column temperature of 30℃.HPLC fingerprints of A.grossedentata and closely related species was established and a combination of chemical pattern recognition methods such as similarity evaluation,cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA),were used to evaluate the quality of A.grossedentata and closely related species from different producing areas.Results The HPLC fingerprint method was developed in accordance with the methodological requirements,with 31 common peaks from 18 batches of A.grossedentata.The peaks 4,9,11,14,15 and 24 were qualitatively identified as gallic acid,catechin,dihydromyricetin,rutin,myricetrin andmyricetin,respectively,and the similarity was more than 0.8.The samples were divided into two categories according to different origins through CA and the results of PCA were consistent with it.Moreover,thirteen to fourteen markers of variation were identified in OPLS-DA,with dihydromyricetin and myricetin as the main markers of variation in the different origins of A.grossedentata and closely related species.Conclusion The established fingerprint method is stable,reliable and reproducible;Combined with chemical pattern recognition,it can be used for the quality evaluation of Ampelopsis genus as raw material plant of vine tea.

关 键 词：藤茶 显齿蛇葡萄 HPLC指纹图谱 化学模式识别 正交偏最小二乘法-判别分析 没食子酸 儿茶素 二氢杨梅素 芦丁 杨梅苷 杨梅素 

分 类 号：R286.2[医药卫生—中药学]