不同粉碎粒度对人参败毒标准...图谱和多指标成分含量的影响_张利恒.pdf

中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 1429 不同粉碎粒度对人参败毒标准汤物理指纹图谱和多指标成分含量的影响 张利恒，陈 新*，邱智东 长春中医药大学 长白山道地药材药效物质重点研究室，吉林 长春 130117 摘 要：目的 建立人参败毒标准汤的物理指纹图谱和多指标成分含量测定方法，考察不同粉碎粒度对人参败毒散煎出物的影响。方法 分别以过 10、24、50 目筛的人参败毒散制备标准汤，测定盐度（S）、溶解性总固体（total dissolved solids，TDS）、电导率（G）、电位（V）、pH 值、动力黏度（）、固含量（SC）、表面张力（）、折光率（n）9 个物理参数，构建其物理指纹图谱，并进行相似度分析（similarity analysis，SA）、层次聚类分析（hierarchical cluster analysis，HCA）、主成分分析（principal component analysis，PCA）；通过 HPLC-DAD 对标准汤中蛇床子素、二氢欧山芹醇当归酸酯、甘草苷、柚皮苷、新橙皮苷的含量进行测定，利用 HCA 法形成指标成分含量聚类分析图，对物理指纹图谱分析结果进行验证，并考察 9 个物理参数与 5 个指标成分的相关性。结果 15 批人参败毒标准汤未出现异常批次，标准汤 S1S5（过 10 目筛）、S6S10（过24 目筛）、S11S15（过 50 目筛）各聚为一类，不同粉碎粒度批次间的物理指纹图谱相似度均小于 0.95，物理参数 TDS、G、V、pH、SC、n 为人参败毒标准汤物理质量的差异性指标，24 目煮散的 TDS、G、SC、均相对较高；指标成分含量 HCA 结果与物理指纹图谱分析结果一致，过 24 目筛的标准汤指标成分含量均高于过 10、50 目筛的煮散；物理指标TDS、G、V、pH、SC、n 与蛇床子素、二氢欧山芹醇当归酸酯、甘草苷、柚皮苷、新橙皮苷的含量之间显著相关。结论 粉碎粒度对人参败毒散煎出物的物理属性和指标成分含量具有明显影响，人参败毒散的最佳粉碎粒度为过 24 目筛，为其质量标准研究和制剂开发提供理论基础。关键词：人参败毒散；粉碎粒度；物理指纹图谱；HPLC-DAD；相似度分析；层次聚类分析；主成分分析；蛇床子素；二氢欧山芹醇当归酸酯；甘草苷；柚皮苷；新橙皮苷 中图分类号：R283.6 文献标志码：A 文章编号：0253-2670(2023)05-1429-10 DOI:10.7501/j.issn.0253-2670.2023.05.009 Effects of different particle sizes on physical fingerprints and contents of multi-index components in Renshen Baidu standard decoction ZHANG Li-heng,CHEN Xin,QIU Zhi-dong Key Laboratory for Effective Substances of Genuine Medicinal Material in Changbai Mountain,Changchun University of Chinese Medicine,Changchun 130117,China Abstract:Objective To establish the physical fingerprints of Renshen Baidu standard decoction and the determination method of multi-index components,and investigate the effect of different crushing particle sizes on the decoction of Renshen Baidu powder.Methods The standard decoction was prepared with Renshen Baidu powder that passed through 10,24,and 50 mesh sieve,and nine physical parameters including salinity(S),total dissolved solids(TDS),conductivity(G),potential(V),pH value,dynamic viscosity(),solid content(SC),surface tension(),refractive index(n)were measured.The physical fingerprints were established,and similarity analysis,hierarchical cluster analysis,and principal component analysis were performed;HPLC-DAD was used to determine the contents of osthol,dihydroeosanthinol-angelate,liquiritin,naringin and neohesperidin in the standard decoction,and the hierarchical cluster analysis method was used to form a cluster analysis chart of the content of the index components,and the results of physical fingerprint analysis were verified.The correlation between nine physical parameters and five index components was investigated.Results There were no abnormal batches of 15 batches of Renshen Baidu standard decoction.The standard decoctions 收稿日期：2022-09-26 基金项目：重大疫情防治中药方剂储备库建设项目（吉中医药发202111 号）作者简介：张利恒（1996），女，硕士研究生，研究方向为中药及复方药效物质基础和质量标准。Tel:17710904859 E-mail:*通信作者：陈 新，博士，教授，从事中药及复方药效物质基础和质量标准研究。Tel:13596000134 E-mail: 1430 中草药 中草药 2023 年年 3 月月 第第 54 卷卷 第第 5 期期 Chinese Traditional and Herbal Drugs 2023 March Vol.54 No.5 S1S5(passed through 10 mesh sieve),S6S10(passed through 24 mesh sieve),and S11S15(passed through 50 mesh sieve)were grouped together.The similarity of physical fingerprints between batches of different crushed particle sizes is less than 0.95,and the physical parameters TDS,G,V,pH,SC,n are the differences in the physical quality of Renshen Baidu standard decoction,The TDS,G,SC,and of Renshen Baidu powder through 24 mesh sieve are relatively high;The cluster analysis results of index component content were consistent with the results of physical fingerprint analysis.The index component content of the standard decoction through 24 mesh sieve was higher than the 10 mesh sieve and 50 mesh sieve;The physical indexes TDS,G,V,pH,SC,n were significantly correlated with the contents of osthol,dihydropicolin-angelate,liquiritin,naringin and neohesperidin.Conclusion The crushing particle size has obvious effect on the physical properties and the content of index component of the decoction of Renshen Baidu powder.The best crushing particle size of Renshen Baidu powder is the drug particle passing through the 24 mesh sieve,which provides a theoretical basis for its quality standard research and preparation development.Key words:Renshen Baidu powder;particle size;physical fingerprint;HPLC-DAD;similarity analysis;hierarchical cluster analysis;principal component analysis;osthol;dihydroeosanthinol-angelate;iquiritin;naringin;neohesperidin 人参败毒散始载于 太平惠民和剂局方，由羌活、独活、柴胡、川芎、桔梗、前胡、茯苓、枳壳、人参、甘草、生姜、薄荷组成1-2，此方兼顾正邪，可补不滞邪，祛邪不伤正，为治疗疫病的经典古方之一3-4。方中化学成分繁多复杂，主要有效成分为香豆素类、黄酮类、皂苷类等化合物。其中，香豆素类成分蛇床子素、二氢欧山芹醇当归酸酯具有较明显的抗炎、镇痛、平喘、抗病毒等药理作用5；黄酮苷类成分新橙皮苷、柚皮苷具有抗氧化、抗炎等药效作用6-7，甘草苷具有扩张外周血管、调节免疫功能等8；皂苷类成分柴胡皂苷 a、b 具有抗炎、解热、抗病毒、增强免疫力的功效9-10。课题组参考了中国药典2020 年版和相关文献报道11-12，进行前期研究，发现人参败毒散中蛇床子素、二氢欧山芹醇当归酸酯、甘草苷、柚皮苷、新橙皮苷的含量较高且稳定。物理指纹图谱法，可以直观反映中药方剂不同批次间各物理指标的相似及差异程度，同时能综合表征标准汤的物理属性，可用于中药方剂的质量评价13。在物理指标中，盐度（S）为液体中盐类物质的质量分数，是与药物吸收相关的影响因素；溶解性总固体（total dissolved solids，TDS）值反映了水溶液中所有固体物质的总量，具体指水中钾、钠、钙等矿物质离子和有机物等；电