Analysis of Pharmacodynamic Material Basis of Qingyan Formula Based on UPLC-Q-Orbitrap-MS and Molecular Docking
10.13422/j.cnki.syfjx.20220353
- VernacularTitle:基于UPLC-Q-Orbitrap-MS和分子对接技术的青盐方药效物质基础分析
- Author:
Zhao ZOU
1
;
Jiashan LI
2
;
Ying XU
2
;
Yibo YANG
2
;
Panyu XU
2
;
Hanqian DU
2
;
Weifeng ZHU
1
;
Na LIN
2
Author Information
1. Jiangxi University of Chinese Medicine，Nanchang 330004，China
2. Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences，Beijing 100700，China
- Publication Type:Journal Article
- Keywords:
Qingyan formula;
serum pharmacochemistry;
molecular docking;
prototype components;
fingerprint;
estrogen receptor （ER）;
ultra-high performance liquid chromatography-quadrupole electrostatic field-orbital trap high resolution mass spectrometry （UPLC-Q-Orbitrap-MS）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(22):159-166
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo study the constituents migrating to blood of Qingyan formula by serum pharmacochemistry， and investigate the binding energy between these constituents and estrogen receptor （ER）， so as to confirm the pharmacodynamic material basis of Qingyan formula in rats. MethodUltra-high performance liquid chromatography-quadrupole electrostatic field-orbital trap high resolution mass spectrometry （UPLC-Q-Orbitrap-MS） was used to determine the constituents migrating to blood of Qingyan formula in rats by comparing the fingerprint differences of 70% ethanol extract of Qingyan formula， 70% ethanol extract of each single drug in this formula， blank serum and serum after administration of 70% ethanol extract of Qingyan formula， according to the retention time， relative molecular weight and the primary and secondary ion fragments provided by MS. Mobile phase was 0.1% formic acid aqueous solution （A）-acetonitrile（B） for gradient elution （0-5 min， 2%-20%B； 5-10 min； 20%-50%B； 10-15 min， 50%-80%B； 15-25 min， 80%-95%B； 25-26 min， 95%-2%B； 26-30 min， 2%B）， the flow rate was 0.3 mL·min-1 and the injection volume was 5 μL， electrospray ionization was used with detection range of m/z 150-2 000， positive and negative ion scanning modes. Molecular docking technology was used to characterize the binding energy of constituents migrating to blood with ERα and ERβ， and to confirm the material basis of this formula. ResultAfter oral administration of Qingyan formula， 30 components were detected in serum， of which 9 were prototype components and 21 were metabolites. Nine prototype components were identified as monotropein， asperuloside， verbascoside， β-ecdysone， allantoin， deacetyl asperuloside acid， echinacoside， betaine and caffeic acid， 21 metabolites mainly included organic acids， amino acids， cholines and so on. The binding energies of the above 9 prototype components with ERα were -6.7， -8.9， -6.0， -5.7， -5.3， -4.9， -7.3， -3.3， -6.3 kcal·mol-1 （1 kcal≈4 184 J）， and the binding energies of them with ERβ were -6.6， -7.2， -7.7， 8.0， -7.4， -5.5， -6.9， -3.6， -6.4 kcal·mol-1， respectively. ConclusionThese nine prototype components into blood are the active ingredients of Qingyan formula that play estrogen-like role in the body， which can provide experimental basis for the formulation of quality standards and subsequent research and development of Qingyan formula.
