A strategy based on liquid-liquid-refining extraction and high-speed counter-current chromatography for the bioassay-guided separation of active compound from Taraxacum mongolicum
| Autor: | Dezhi Huang, Dongyu Gu, Yunxiao Wang, Jing Tian, Jinlong Han, Yi Yang, Xuan Zhao, Wenqiang Zeng |
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| Rok vydání: | 2020 |
| Předmět: |
Magnetic Resonance Spectroscopy
Taraxacum Liquid-Liquid Extraction Ethyl acetate Fraction (chemistry) Acetates 010402 general chemistry 01 natural sciences Biochemistry Chemistry Techniques Analytical Analytical Chemistry Hydrophobic effect Inhibitory Concentration 50 chemistry.chemical_compound Countercurrent chromatography Luteolin Countercurrent Distribution Chromatography Plant Extracts 010401 analytical chemistry Organic Chemistry Extraction (chemistry) General Medicine Carbon-13 NMR 0104 chemical sciences Enzyme Activation Molecular Docking Simulation chemistry Proton NMR Biological Assay alpha-Amylases |
| Zdroj: | Journal of Chromatography A. 1614:460727 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/j.chroma.2019.460727 |
| Popis: | The research of natural active substances is facing the problems of low separation efficiency and active component loss due to the complex composition of natural extracts. In this study, a strategy based on liquid-liquid-refining extraction and high-speed counter-current chromatography was established to solve this problem. Separation of an active compound with the α-amylase inhibitory activity from Taraxacum mongolicum Hand. -Mazz. was presented as an example. The ethyl acetate extract (FA) from T. mongolicum exhibited the potential effect on α-amylase and was divided into 8 fractions (FB-FI) by liquid-liquid-refining extraction. The results showed that the activity of FE was higher than the others. According to the results of liquid-liquid-refining extraction, a two-phase solvent system with a slightly higher polarity was selected to separate the fraction by HSCCC, and 110 mg of compound was separated from 900 mg FA using the model of consecutive separation. The compound was identified as luteolin by 1H NMR and 13C NMR. The IC50 of luteolin against α-amylase was 42.33±0.82 μg/mL. Then, molecular docking was introduced to study the relationship between the activity and the structure. The results showed that luteolin enfolded in the catalytic site of α-amylase through hydrogen bonds, van der Waals force and hydrophobic interaction, thus inhibiting the activity of the enzyme. |
| Databáze: | OpenAIRE |
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