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 Chengdu 611137,China.
College of Medicine,Southwest Jiaotong University Chengdu 610031,China.
Zhongguo Zhong Yao Za Zhi. 2019 Dec;44(23):5151-5158. doi: 10.19540/j.cnki.cjcmm.20190916.304.
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.
基于古代九蒸九晒法,研究了何首乌在不同炮制过程中的化学成分变化和质量特征。采用黑豆汁九蒸九晒法和药典方法测定其白藜芦醇苷、蒽醌类和多糖的含量。建立了HPLC化学指纹图谱,并使用SIMCA 14.1软件对不同炮制品进行正交偏最小二乘法判别分析(OPLS-DA),以评价样品间的质量差异。含量测定结果表明,在何首乌不同炮制品及药典品中,随着炮制次数和蒸制时间的增加,白藜芦醇苷和结合蒽醌呈下降趋势,游离蒽醌、总蒽醌和多糖呈上升趋势。六蒸六晒品可作为经典蒸制的终点。指纹图谱结果表明,何首乌在不同炮制过程中的化学成分存在显著差异。可鉴定出白藜芦醇苷(峰13)、大黄素(峰21)和大黄素甲醚(峰24)。通过对样品归一化后26个色谱峰的相对峰面积进行比较(以峰7为参照),发现炮制品中有12个峰的相对峰面积高于生品,13个峰降低;根据OPLS-DA统计分析,不同炮制程度的何首乌进一步分为三类,样品S1为I类,S2-S5为II类,S6-S11为III类。VIP值高于1.0的8个峰依次为峰13、21、4、3、11、14、5和峰24。这8种化学成分是区分何首乌九蒸九晒过程中差异的主要成分,表明以白藜芦醇苷、大黄素和大黄素甲醚作为何首乌质量控制指标具有合理性。本实验建立的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的方法,明确了六蒸六晒作为改进的经典炮制工艺,并更清晰地界定了何首乌九蒸九晒过程中化学成分的整体动态变化。为何首乌不同炮制品的化学质量评价模型提供了依据。