天然产物研究与开发 ›› 2022, Vol. 34 ›› Issue (6): 941-953.doi: 10.16333/j.1001-6880.2022.6.005

• 研究论文 • 上一篇    下一篇

基于指纹图谱及体外抗氧化活性试验的灯盏花标准汤剂的质量评价

李艳荣1,2，杜义龙1,2，赵胜男1,2，王子怡1,2，周维维1,2,3，潘海峰1,2*   

  1. 1承德医学院中药研究所；2河北省中药研究与开发重点实验室；3承德市食品药品检验检测中心，承德 067000
  • 出版日期:2022-06-28 发布日期:2022-06-30
  • 基金资助:
    河北省中药研究与开发重点实验室开放课题（ZYKF2020009）；河北省高校重点学科建设项目（冀教高［2013］4）

Quality evaluation of Erigerontis Herba standard decoction based on fingerprint and antioxidant activity test in vitro

LI Yan-rong1,2,DU Yi-long1,2,ZHAO Sheng-nan1,2,WANG Zi-yi1,2,ZHOU Wei-wei1,2,3,PAN Hai-feng1,2*   

  1. 1Institute of Traditional Chinese Medicine,Chengde Medical University;2Hebei Province Key Laboratory of Research and Development for Chinese Medicine;3Chengde Food and Drug Inspection and Testing Center,Chengde 067000,China
  • Online:2022-06-28 Published:2022-06-30

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摘要：

运用HPLC法测定灯盏花标准汤剂的指纹图谱，测定其体外抗氧化活性，采用双变量相关分析及化学计量学方法对灯盏花标准汤剂进行质量评价。以1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除率、2,2′-连氨-（3-乙基苯并噻唑啉-6-磺酸）二氨盐（ABTS）自由基清除率为抗氧化作用的药效指标测定其抗氧化能力。采用紫外-可见分光光度法总黄酮的含量；高效液相色谱法建立灯盏花标准汤剂的指纹图谱；运用双变量相关性分析筛选出与DPPH和ABTS自由基半数抑制率（IC50）显著相关的分析变量，进一步通过聚类分析和主成分分析对灯盏花标准汤剂进行质量评价。灯盏花标准汤剂DPPH的IC50值为1.00~3.39 mg/mL，对ABTS的IC50值为0.254~2.16 mg/mL；总黄酮的含量为2.85~11.3 mg/mL，IC50值与总黄酮的含量有显著的相关性（r=-0.718，r=-0.841）；指纹图谱的相似度0.770~0.993，共得到21个共有峰，用对照品指认了8个已知成分；双变量相关性分析共筛选出与DPPH的IC50、ABTS的IC50显著相关的共有峰分别为10、13个；聚类分析（CA）和主成分分析（PCA）将样品分成2类，建立主成分评分模型FDPPH=0.719 9F1+0.112 3F2+0.099 27F3，FABTS=0.700 5F1+0.131 9F2+0.086 57F3。在CA和PCA的基础上进一步采用OPLS-DA，找出了峰11、13、14、15是与清除DPPH自由基相关的主要差异峰；峰11、15、14、8是与清除ABTS自由基相关的主要差异峰。本文以“谱-效”结合的思路评价灯盏花标准汤剂的质量，可为灯盏花及其制剂的质量控制提供依据。

关键词: 灯盏花, 标准汤剂, 指纹图谱, 抗氧化活性, 质量评价

Abstract:

HPLC method was used to determine the fingerprint of Erigerontis Herba standard decoction.Its antioxidant activity in vitro was determined.The quality of Erigerontis Herba decoction was evaluated by bivariate correlation analysis and stoichiometry.The scavenging rate of 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radical and 2,2′-hydrazine - (3-ethylbenzothiazoline-6-sulfonic acid) diammonia salt (ABTS) free radical were used as pharmacodynamic indexes to determine the antioxidant capacity.The content of total flavonoids was determined by UV-visible spectrophotometry.The fingerprint of Erigerontis Herba standard decoction was established by HPLC.Bivariate correlation analysis was used to screen out the analysis variables that were significantly correlated with DPPH and ABTS radical 50% inhibition rate (IC50).Further,cluster analysis (CA) and principal component analysis (PCA) were used to evaluate the quality of Erigerontis Herba standard decoction.The IC50 values of Erigerontis Herba standard decoction DPPH were 1.00~3.39 mg/mL and 0.254~2.16 mg/mL for ABTS.The total flavonoid content was 2.85~11.3 mg/mL,and the IC50 value was significantly correlated with the total flavonoid content (r=-0.718,r=-0.841).The similarity of fingerprints was 0.770~0.993,21 common peaks were obtained,and 8 known components were identified with reference substance. A total of 10 and 13 common peaks were found to be significantly correlated with IC50 of DPPH and ABTS by bivariate correlation analysis.Cluster analysis and principal component analysis divided the samples into two categories and established the principal component scoring model.FDPPH=0.719 9F1+0.112 3F2+0.099 27F3,FABTS=0.700 5F1+0.131 9F2+0.086 57F3.Based on CA and PCA,OPLS-DA was further used to find that peaks 11,13,14 and 15 were the main difference peaks related to DPPH free radical scavenging.Peaks 11,15,14 and 8 were the main difference peaks associated with ABTS free radical scavenging.This paper evaluated the quality of Erigerontis Herba standard decoction by combining spectrum and efficiency,which could provide basis for the quality control of Erigerontis Herba and its preparations.

Key words: 灯盏花, 标准汤剂, 指纹图谱, 抗氧化活性, 质量评价

中图分类号:  R284.2

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