中国中药杂志

2016, v.41(20) 3773-3781

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中药三七质量评价半微量方法及在三七连作障碍样品分析中的应用
A semimicroquality evaluation method on Panax notoginseng and its application in analysis of continuous cropping obstacles research samples

曹怡;王超群;徐风;贾秀虹;刘广学;杨生超;龙光强;陈中坚;魏富刚;杨绍周;福田浩三;王璇;蔡少青;
CAO Yi;WANG Chao-qun;XU Feng;JIA Xiu-hong;LIU Guang-xue;YANG Sheng-chao;LONG Guang-qiang;CHEN Zhong-jian;WEI Fu-zhou;YANG Shao-zhou;FUKUDA Kozo;WANG Xuan;CAI Shao-qing;Department of Chemical Biology,School of Pharmaceutical Sciences,Peking University;State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences,Peking University;Yunnan Provincial Research Center on Good Agricultural Practice for Dominant Chinese Medicinal Materials,Yunnan Agricultural University;WenshanS

摘要(Abstract):

该研究旨在建立三七质量评价半微量方法并应用于三七连作障碍样品分析。采用100倍量水饱和正丁醇超声提取0.1 g三七样品,所建立的半微量提取方法相比于甲醇常量提取法对5个主要皂苷类成分及三七皂苷K和Fa提取效率高9.6%~20.6%,且重复性良好,RSD<2.5%。LC-MS-IT-TOF技术指认指纹图谱16个皂苷类成分特征峰,包括8个人参二醇型皂苷和8个人参三醇型皂苷。利用HPLC指纹图谱相似度评价及对特征峰峰面积提取可进行三七的质量分析。应用所建立的方法对人工干预克服连作障碍三七样品质量评价的结果表明:指纹图谱相似度、三七皂苷Fa的相对含量及三七皂苷K与三七皂苷Fa的比值(N-K/Fa值)均可作为评价连作地三七质量恢复程度的指标,其中N-K/Fa值同样适用于不同生长年限三七样品的区分。正常(无连作栽培)组样品三七皂苷K与三七皂苷Fa的峰面积比值为0.45~1.33;连作组样品个头小于正常组,指纹图谱与正常组相似度均低于0.87,三七皂苷Fa的峰面积显著低于正常组,N-K/Fa值显著高于正常组,为2.35~4.74,与正常组三七样品质量存在差异。人工干预一组的样品与正常组相似度均高于0.87,且16个特征峰的峰面积和分布模式均与正常组无显著差异,N-K/Fa值为0.42~2.06。人工干预二组的样品除重量较小的样品外,其余均与正常组指纹图谱相似度较高,三七皂苷K显著高于正常组,N-K/Fa值为0.96~6.16。连作障碍经人工干预后三七的质量均有不同程度的恢复和提高,其中人工干预一组的三七质量恢复情况优于人工干预二组。该研究建立的三七质量评价半微量方法,采用指纹图谱及峰面积比值作为评价指标可避免含量测定的繁琐及昂贵,为连作障碍三七样品的质量分析提供了简便、快速和准确的评价方法,且可应用于三七不同年限等样品的质量评价。
Panax notoginseng is a commonly used traditional Chinese medicine with blood activating effect while has continuous cropping obstacle problem in planting process. In present study,a semimicroextraction method with water-saturated n-butanol on 0. 1 g notoginseng sample was established with good repeatability( RSD < 2. 5 %) and 9. 6 %-20. 6 % higher extraction efficiency of seven saponins than the conventional method. A total of 16 characteristic peaks were identified by LC-MS-IT-TOF,including eight 20( S)-protopanaxatriol( PPT) type saponins and eight 20( S)-protopanaxadiol( PPD) type saponins. The established method was utilized to evaluate the quality of notoginseng samples cultivated by manual intervened methods to overcome continuous cropping obstacles. As a result,HPLC fingerprint similarity,content of Fa and ratio of notoginsenoside K and notoginsenoside Fa( N-K / Fa) were found out to be as valuatable markers of the quality of samples in continuous cropping obstacle research,of which N-K / Fa could also be applied to the analysis of notoginseng samples with different growth years. Notoginseng samples with continuous cropping obstacle had HPLC fingerprint similarity lower than 0. 87,in consistent with normal sample,and had significant lower content of notoginsenoside Fa and significant higher N-K / Fa( 2. 35-4. 74) than normal group( 0. 45-1. 33). All samples in the first group with manual intervention showed high similarity with normal group( > 0. 87),similar content of common peaks and N-K / Fa( 0. 42-2. 06). The content of notoginsenoside K in the second group with manual intervention was higher than normal group. All samples except two displayed similarity higher than 0. 87 and possessed content of 16 saponins close to normal group. The result showed that notoginseng samples with continuous cropping obstacle had lower quality than normal sample. And manual intervened methods could improve their quality in different levels. The method established in this study was simple,fast and accurate,and the markers may provide new guides for quality control in continuous cropping obstacle research of notoginseng.

关键词(KeyWords): 三七;半微量高效提取;连作障碍;LC-MS-IT-TOF;HPLC指纹图谱;特征峰;三七皂苷K/三七皂苷Fa
Panax notoginseng;semimicro and highly efficient extraction method;continuous cropping obstacles;LC-MS-IT-TOF;HPLC fingerprint;characteristic peaks;notoginsenoside K/notoginsenoside Fa

Abstract:

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基金项目(Foundation): 国家“重大新药创制”科技重大专项(2014ZX09304307-001-012);; 国家中医药管理局中医药行业科研专项(201407003)

作者(Author): 曹怡;王超群;徐风;贾秀虹;刘广学;杨生超;龙光强;陈中坚;魏富刚;杨绍周;福田浩三;王璇;蔡少青;
CAO Yi;WANG Chao-qun;XU Feng;JIA Xiu-hong;LIU Guang-xue;YANG Sheng-chao;LONG Guang-qiang;CHEN Zhong-jian;WEI Fu-zhou;YANG Shao-zhou;FUKUDA Kozo;WANG Xuan;CAI Shao-qing;Department of Chemical Biology,School of Pharmaceutical Sciences,Peking University;State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences,Peking University;Yunnan Provincial Research Center on Good Agricultural Practice for Dominant Chinese Medicinal Materials,Yunnan Agricultural University;WenshanS

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