天然产物研究与开发 ›› 2022, Vol. 34 ›› Issue (8): 1301-1310.doi: 10.16333/j.1001-6880.2022.8.004

• 研究论文 • 上一篇    下一篇

UPLC指纹图谱结合多元统计分析的吴茱萸与制吴茱萸成分变化研究

杨文惠1,2,吕渭升1,2,孙冬梅1,3,罗文汇1,2,杨   洁1,2,何民友1,2,潘礼业1,2,李振雨1,2*   

  1. 1广东一方制药有限公司;2广东省中药配方颗粒企业重点实验室,佛山 528244;3江西一方天江药业有限公司,南昌 330000
  • 出版日期:2022-08-28 发布日期:2022-09-08
  • 基金资助:
    江西省中医药标准化项目(2020A10);湖南省自然科学基金-科药联合基金(2021JJ80057)

Study on the changes of components in Euodiae Fructus and prepared Euodiae Fructus by UPLC fingerprint combined with multivariate statistical analysis

YANG Wen-hui1,2,LYU Wei-sheng1,2,SUN Dong-mei1,3,LUO Wen-hui1,2,YANG Jie1,2,HE Min-you1,2,PAN Li-ye1,2,LI Zhen-yu1,2*
  

  1. 1GuangdongYifang Pharmaceutical Co.,Ltd.China;2Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Formula,Foshan 528244,China;3Jiangxi Yifang Tianjiang Pharmaceutical Co.,Ltd.,Nanchang 330000,China
  • Online:2022-08-28 Published:2022-09-08

摘要:

建立吴茱萸及制吴茱萸指纹图谱,并结合多元统计分析吴茱萸与制吴茱萸中化学成分的变化。采用超高效液相色谱(UPLC)技术。色谱柱为YMC Triart C18(100 mm×2.1 mm,1.9 μm),流动相为乙腈-0.1%磷酸水(梯度洗脱),流速为0.30 mL/min,柱温为30 ℃,检测波长为254 nm,进样量为1 μL。建立吴茱萸与制吴茱萸UPLC指纹图谱,并采用相似度计算和正交偏最小二乘法判别分析(OPLS-DA)对指纹图谱进行分析评价,以变量重要性投影(VIP)值大于1为标准筛选吴茱萸炮制前后的差异性成分。以平均峰面积为变量,对吴茱萸和制吴茱萸差异成分色谱峰的平均峰面积进行配对t检验。吴茱萸指纹图谱共标识出27个共有指纹峰,指认了其中8个化学成分,分别为新绿原酸、绿原酸、隐绿原酸、咖啡酸、金丝桃苷、去氢吴茱萸碱、吴茱萸碱和吴茱萸次碱。制吴茱萸经炮制后指纹图谱新增甘草酸色谱峰。相似度结果显示,江西产区相似度较低;湖南产区次之;广西产区较高。正交偏最小二乘法判别分析(OPLS-DA)共找到6个差异性标志物,差异显著性排序分别为甘草酸>吴茱萸碱>峰3>峰1>绿原酸>峰2。除甘草酸为新增成分外,吴茱萸炮制后其余5种化学成分的含量均有一定程度降低。吴茱萸炮制后,峰21(吴茱萸碱)平均峰面积明显降低,P<0.05,峰28(甘草酸)为新增成分,其余5个差异性化学成分的平均峰面积均有不同程度的降低,但无明显统计学意义。本分析方法专属性强,重复性好,初步探讨了吴茱萸与制吴茱萸药理作用与化学成分的相关性,一定程度上阐释了吴茱萸炮制前后药理作用改变与化学成分变化之间的联系,可为吴茱萸和制吴茱萸质量及炮制研究提供数据参考。

关键词: 吴茱萸, 制吴茱萸, UPLC指纹图谱, 相似度, 正交偏最小二乘判别分析

Abstract:

To establish the fingerprints of Euodiae Fructus and prepared Euodiae Fructus,the changes of chemical components in Euodiae Fructus and its preparation were analyzed by multivariate statistics. Using ultra high performance liquid chromatography (UPLC) technology. Chromatographic column was YMC Triart C18 (100 mm×2.1 mm,1.9 μm).The mobile phase consisted of acetonitrile-0.1% phosphoric acid water (gradient elution),the flow rate was 0.30 mL/min,the column temperature was 30 ℃,the detection wavelength was 254 nm,the injection volume was 1 μL.The UPLC fingerprints of Euodiae Fructus and its preparation were established,and the similarity calculation and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to analyze and evaluate the fingerprints,the variable importance projection (VIP) value greater than 1 was used as the criterion to screen the differential components before and after processing. Taking the average peak area as the variable,the average peak area of the chromatographic peaks of different components of Euodiae Fructus and prepared Euodiae Fructus was tested by paired t-test. A total of 27 common fingerprint peaks were identified in the fingerprint of Euodiae Fructus,Eight chemical components were identified as neocrogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,hyperin,dehydroevotaine,rutaecarine and rutaecarine. Glycyrrhizic acid chromatographic peak was added to the fingerprint of processed Euodiae Fructus. The results of similarity degree showed that the similarity degree of Jiangxi production area was low. Hunan production area was lower. Guangxi production area was relatively high. Orthogonal partial least squares discriminant analysis (OPLS-DA) found six markers of difference,and the order of significance of difference was glycyrrhizic acid > evocorine > peak 3> peak 1> chlorogenic acid > peak 2. Except that glycyrrhizic acid is a new component,the contents of the other five chemical components of Euodiae Fructus after processing are reduced to a certain extent. After the processing of Euodiae Fructus,the average peak area of peak 21 (evodiamine) decreased significantly (P<0.05),peak 28 (glycyrrhizic acid) was a new component,and the average peak areas of the other five differential chemical components decreased in varying degrees,but there was no significant statistical significance. This analysis method has strong specificity and good repeatability. It preliminarily discusses the correlation between the pharmacological effects and chemical components of Euodiae Fructus and prepared Euodiae Fructus,and explains the relationship between the changes of pharmacological effects and chemical components before and after Euodiae Fructus processing to a certain extent,which can provide data reference for the quality and processing research of Euodiae Fructus and prepared Euodiae Fructus.

Key words: Euodiae Fructus, prepared Euodiae Fructus, UPLC fingerprint, similarity, orthogonal partial least squares discriminant analysis

中图分类号:  R282.4