Spectrum-effect Relationship Analysis Between HPLC Fingerprint and Immunomodulatory Activity of Zhenqi Fuzheng Granules
10.13422/j.cnki.syfjx.20211853
- VernacularTitle:贞芪扶正颗粒HPLC指纹图谱与其增强免疫功能的谱效关系分析
- Author:
Wen-yue JIANG
1
;
Shu-li HAN
1
;
Ming-zhe TANG
1
;
Gui-fang ZHAO
1
;
Lu GAO
2
;
Jing-wei MIAO
1
;
Peng-dong LI
1
Author Information
1. The Sixth Affiliated Hospital of Guangzhou Medical University， Qingyuan People′s Hospital，Qingyuan 511518，China
2. Jilin Modern Chinese Medicine Engineering and Research Center Co. Ltd.，Changchun 130012，China
- Publication Type:Journal Article
- Keywords:
Zhenqi Fuzheng granules;
immune regulation;
high performance liquid chromatography （HPLC）;
fingerprint;
mass spectrometry （MS）;
chromatographic pharmacodynamics;
material basis
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(8):123-130
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the pharmacodynamic ingredients of Zhenqi Fuzheng granules （ZFG） for immunomodulatory through spectrum-effect relationship analysis， which provides experimental basis for improving the quality standard of ZFG. MethodEighteen batches of ZFG from six manufacturers were collected for analysis. The fingerprints were established by high performance liquid chromatography （HPLC）. Acetonitrile （A）-0.1% formic acid aqueous solution （B） were adopted as the mobile phase with gradient elution （0-15 min， 5%A； 15-23 min， 5%-8%A； 23-30 min， 8%-11%A； 30-45 min， 11%-18%A； 45-60 min， 18%-21%A； 60-67 min， 21%-23%A； 67-90 min， 23%-37%A）， the detection wavelength was 220 nm. Chemometric analysis such as similarity analysis and hierarchical cluster analysis （HCA） were subsequently used to analyze the similarities and chemical differences among these samples. A cyclophosphamide-induced immunodeficiency mouse model was used to evaluate the immune-enhancing effects of the products from different manufacturers. The spectrum-effect relationship between HPLC fingerprints and the immunomodulatory effects was examined using Spearman bivariate correlation analysis. HPLC coupled with mass spectrometry （HPLC-MSn） was used to identify the spectrum-effect related peaks with electrospray ionization， positive and negative ion modes， and scanning range of m/z 100-1 500. ResultThe HPLC fingerprint of ZFG was established， and twenty peaks with good resolution were selected as common peaks. The results of quality analysis and pharmacodynamic test showed there were significant differences in both ingredients content and immune-enhancing effects of ZFG from different manufacturers. Through spectrum-effect relationship study， twelve peaks were screened as bioactive ingredients peaks. Thereafter， eight peaks among them were subsequently identified by HPLC-MSn. They were salidroside （peak 2）， echinacoside （peak 5）， calycosin-7-glucoside （peak 6）， isomer of specnuezhenide （peak 7）， isonuezhenide （peak 9）， calycosin （peak 11）， nuezhenide G13 or oleonuezhenide （peak 14）， and formononetin （peak 18）， respectively. ConclusionThere are differences in quality and efficacy of ZFG produced by different manufacturers. Through spectrum-effect relationship analysis， the medicinal ingredients of ZFG for immune-enhancing effects are screened， which can provide reference for the improvement of its quality standard.
