成颜芬;聂欣;谭睿;张良;吴亿晗;章津铭;傅超美;
CHENG Yan-fen;NIE Xin;TAN Rui;ZHANG Liang;WU Yi-han;ZHANG Jin-ming;FU Chao-mei;College Pharmacy,Chengdu University of Traditional Chinese Medicine;College of Medicine,Southwest Jiaotong University;

• 1:成都中医药大学药学院
• 2:西南交通大学医学院

摘要(Abstract)：

遵循古法九蒸九晒,探究何首乌不同炮制过程中化学成分的变化规律和质量概貌。采用传统经典黑豆汁九蒸九晒法和现行版药典法进行炮制,测定不同炮制程度何首乌成品中二苯乙烯苷、蒽醌、多糖含量;并建立HPLC化学指纹图谱,采用SIMCA 14. 1软件对不同炮制成品进行正交偏最小二乘法-判别分析(OPLS-DA),以评价各样品之间的质量差异。含量测定结果表明,随着炮制次数和蒸晒时间的增加,九蒸九晒炮制过程中的何首乌和药典品中二苯乙烯苷、结合蒽醌总体呈现降低趋势,游离蒽醌、总蒽醌、多糖呈现上升趋势,且发现六蒸六晒品作为何首乌九蒸九晒法的炮制终点。指纹图谱结果表明,不同炮制过程何首乌之间化学成分存在显著差异,指认出二苯乙烯苷(13号峰),大黄素(21号峰),大黄素甲醚(24号峰),通过比较样品中26个色谱峰经归一化处理后的相对峰面积(参照峰为7号峰),发现炮制品中12个色谱峰的相对峰面积较于生品升高,13个色谱峰则降低;结合相对峰面积的OPLS-DA统计分析,可进一步将不同炮制程度的何首乌样品分为三类,样品S1为Ⅰ类,S2～S5为Ⅱ类,S6～S11为Ⅲ类,且VIP大于1. 0的有8个色谱峰,按大小依次为13,21,4,3,11,14,5,24号色谱峰,该8种化学成分是区分九蒸九晒炮制过程中何首乌样品之间差异的主要成分,同时说明将二苯乙烯苷、大黄素、大黄素甲醚作为何首乌质量控制指标具有合理性。该文方法符合方法学验证要求,建立了多成分含量测定结合指纹图谱方法,明确六蒸六晒法作为何首乌经典炮制工艺的改良,更加清晰地掌握何首乌在九蒸九晒炮制中化学成分的动态变化全过程,为九蒸九晒法何首乌的化学质量评价模式提供依据。
Based on the ancient method of nine-steaming and nine-sun-curing,the chemical composition changes and quality profiles in different processes of Polygoni Multiflori Radix were studied. Their contents of stilbene glycoside,anthraquinones and polysaccharides were determined by nine-steaming and nine-sun-curing with black bean juice and pharmacopoeia method. HPLC chemical fingerprints were established,and orthogonal partial least squares-discriminant analysis( OPLS-DA) was performed on different processed products using SIMCA 14. 1 software to evaluate the quality difference between samples. The results of content determination show that,with the increase of the number of processing and steaming times,the stilbene glycoside and the combined anthraquinone showed a decreasing trend,and the free anthraquinone,total anthraquinone and polysaccharide showed an upward trend in the different preparations of Polygoni Multiflori Radix and Pharmacopoeia. Six-steamed and six-sun-cured products can be used as the finishing point for the classic steaming. Fingerprint results showed that there were significant differences in chemical composition in Polygoni Multiflori Radix at different processing processes. It can be identified stilbene glycoside( peak 13),emodin( peak 21),and physcion( peak 24). By comparing the relative peak areas of the 26 chromatographic peaks in the sample after normalization( the reference is peak 7),it was found that the relative peak areas of 12 peaks in the processed products were higher than the raw products,13 peaks were reduced; according to statistical analysis of OPLS-DA,Polygoni Multiflori Radix at different processing degrees was further divided into three categories,sample S1 was class I,S2-S5 were class Ⅱ,and S6-S11 were class Ⅲ. And 8 peaks with the VIP value higher than 1. 0 were peak 13,21,4,3,11,14,5,and 24 in order. The eight chemical components were the main components to distinguish the difference between Polygoni Multiflori Radix in the process of nine-steaming and nine-sun-curing,suggesting that it was rational to use stilbene glycoside,emodin and emodin methyl ether as quality control indicators of Polygoni Multiflori Radix. The method established in this experiment conformed to the methodological verification requirements,established a method of multi-component content determination combined with fingerprint,and clarified that six-steaming and six-sun-curing was used as an improved classical processing technology,and more clearly defined the whole dynamic change of chemical composition in Polygoni Multiflori Radix by nine-steaming and ninesun-curing process. It provides a basis for the chemical quality evaluation model about different processed products of Polygoni Multiflori Radix.

关键词(KeyWords)： 何首乌;九蒸九晒;二苯乙烯苷;蒽醌;多糖;含量测定;化学指纹图谱;正交偏最小二乘法-判别分析
Polygoni Multiflori Radix;nine-steaming and nine-sun-curing;stilbene glycoside;anthraquinones;polysaccharides;content determination;chemical fingerprint;OPLS-DA

Abstract:

Keywords:

基金项目(Foundation): 中国科协第四届青年人才托举工程项目(2018QNRC1-01)

作者(Author): 成颜芬;聂欣;谭睿;张良;吴亿晗;章津铭;傅超美;
CHENG Yan-fen;NIE Xin;TAN Rui;ZHANG Liang;WU Yi-han;ZHANG Jin-ming;FU Chao-mei;College Pharmacy,Chengdu University of Traditional Chinese Medicine;College of Medicine,Southwest Jiaotong University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20190916.304

参考文献(References)：

• [1]梁鹂，郑金生，赵中振．何首乌考辨[J]．中国中药杂志，2016,41(23):4456.
• [2]吴成胜，孙蓉．何首乌临床研究进展与安全应用思考[J]．中国中药杂志，2017,42(2):259.
• [3]王浩，杨健，周良云，等．何首乌化学成分与药理作用研究进展[J]．中国实验方剂学杂志，2019,25(13):192.
• [4]赵梦杰，龚小红，党珏，等．炮制时间对何首乌16个成分含量变化影响的研究[J]．中国中药杂志，2017,42(7):1344.
• [5] Lin L,Ni B,Lin H,et al. Traditional usages,botany,phytochemistry,pharmacology and toxicology of Polygonum multiflorum Thunb:a review[J]. J Ethnopharmacol,2015,15(159):158.
• [6]崔鹤蓉，柏兆方，宋海波，等．从古今炮制方法演变探讨何首乌毒性的潜在影响因素[J]．中国中药杂志，2016,41(2):333.
• [7]赵晖，苗艳艳，苗明三．中药九蒸九晒的分析与思考[J]．时珍国医国药，2018,29(11):2713.
• [8]李时珍．本草纲目[M]．刘衡如校注．北京:人民卫生出版社，1975:1288.
• [9]杨文宇，陈祥贵，石泓，等．何首乌九蒸九晒炮制过程中质量指标的变化[J]．时珍国医国药，2015,26(10):2414.
• [10] Meng Y K,Li C Y,Li R Y,et al. Cis-stilbene glucoside in Polygonum multiflorum induces immunological idiosyncratic hepatotoxicity in LPS-treated rats by suppressing PPAR-γ[J]. Acta Pharmacol Sin,2017,38(10):1340.
• [11]杨磊，张曼华，陈文明，等．基于经典与现代炮制方法对何首乌主要化学成分变化规律研究[J]．中华中医药杂志，2018,33(2):770.
• [12]中国药典．一部[S]. 2015:175.
• [13]杨磊，刘梦娇，陈文明，等．2种炮制方法对何首乌中6种成分变化的影响[J]．中成药，2019,41(1):119.
• [14]李旻，吴彬，黄丽凤．何首乌与制何首乌中多糖及水溶性总糖的含量测定[J]．四川中医，2010,28(10):52.
• [15]郑英，李玮，赵贵，等．基于过程控制的何首乌产地加工与炮制一体化方法分析[J]．中国实验方剂学杂志，2018,24(15):29.
• [16]肖榕，林艳，雷思敏，等．UPLC指纹图谱结合模式识别分析不同产地生/制何首乌[J]．中国中药杂志，2017,42(12):2305.
• [17]林艳，肖榕，尹林子，等．基于粗糙集理论构建“谱毒”关系研究何首乌特异质肝毒性物质基础[J]．中国中药杂志，2019,44(3):509.
• [18]严维花，曹虹虹，郭爽，等．当归不同炮制品的UPLC指纹图谱与多成分含量测定研究[J]．中国中药杂志，2019,44(12):2499.
• [19]郜丹，李晓菲，尹萍，等．基于炮制减毒思想的何首乌肝毒性物质基础初步研究[J]．中草药，2017,48(10):2044.