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
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
复制标题
基于液-液精制萃取和高速逆流色谱的策略用于生物测定引导下蒲公英活性成分的分离
DOI：
10.1016/j.chroma.2019.460727
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发表时间：
2020
影响因子：
4.1
通讯作者：
Gu Dongyu
中科院分区：
文献类型：
--
作者：
Yang Yi;Wang Yunxiao;Zeng Wenqiang;Tian Jing;Zhao Xuan;Han Jinlong;Huang Dezhi;Gu Dongyu
天然活性物质的研究由于天然提取物组成复杂，面临着分离效率低、活性成分损失等问题。本研究建立了一种基于液-液-精制萃取和高速逆流色谱的方法来解决这一问题。蒲公英中α-淀粉酶抑制活性成分的分离- 马兹作为一个例子。采用乙酸乙酯萃取法从T.结果表明，蒙古黄芪对α-淀粉酶有较好的影响，经液-液-液精制提取，可将其分为8个组分（FB-FI）。结果表明，FE的活性最高。根据液-液-精制萃取的结果，选择极性稍强的两相溶剂体系，采用HSCCC进行分离，采用连续分离模型，从900 mg FA中分离出110 mg化合物。经1H NMR和13 C NMR确证为木犀草素。木樨草素对α-淀粉酶的IC_（50）为42.33±0.82 μg/mL。然后，利用分子对接的方法研究了其活性与结构的关系。结果表明，木犀草素通过氢键、货车范德华力和疏水相互作用包合在α-淀粉酶的催化位点，从而抑制了酶的活性。
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 fromTaraxacum mongolicumHand. -Mazz. was presented as an example. The ethyl acetate extract (FA) fromT. mongolicumexhibited 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 by1H NMR and13C NMR. The IC50of 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.