马齿苋中抗炎活性物质的提取、分离及结构鉴定

Extraction, Separation and Structural Identification of Anti-inflammatory Active Substances from Purslane (Portulaca oleracea L.)

摘要:

以活性物质示踪为导向，建立脂多糖诱导的RAW264.7巨噬细胞炎症模型对马齿苋中的抗炎物质进行跟踪，采用柱层析提取法、硅胶柱色谱分离法、制备液相色谱法及气相色谱- 质谱联用技术对抗炎物质进行提取分离和结构鉴定。结果表明，石油醚-乙醇、无水乙醇和纯水溶剂依次对马齿苋样品进行提取，三种粗提物将细胞中一氧化氮（Nitric Oxide，NO）的分泌量分别减少至33.13、25.83和20.53 μmol/L，其中石油醚相粗提物的抑制效果最强（P<0.05）。对石油醚相进一步分离得到四个组分，Fr.1、Fr.2和Fr.3组分具有较强的抗炎效果，但Fr.1 和Fr.2 组分含有潜在的毒性成分，选择Fr.3组分继续分离。Fr.3组分经硅胶柱分离得到三个组分，Fr.3.1组分表现出最强的抑制NO的分泌量效果（11.80 μmol/L）。经制备液相色谱进一步纯化及气质分析，确定Fr.3.1组分的主要成分为硬脂酸（47.09%）、邻苯二甲酸二(2-乙基己)酯（13.21%）和其他成分。该研究建立了一种从马齿苋中分离纯化出抗炎物质方法，为马齿苋的开发利用提供理论参考。

Abstract:

To track the anti-inflammatory substances in purslane, the lipopolysaccharide-induced RAW264.7 macrophage inflammation model was established, which was guided by the tracer of active substances. The extraction, separation and structural identification of anti-inflammatory substances in purslane were performed by column chromatography (for extraction), silica gel column chromatography (for separation), and preparative high performance liquid chromatography and gas chromatography-mass spectrometry (for analyses). The results showed that the three crude extracts obtained from purslane through sequential extractions with petroleum ether-ethanol, anhydrous ethanol and pure water solvents reduced the secretion of nitric oxide (NO) in the cells to 33.13, 25.83 and 20.53 μmol/L, respectively, with the crude petroleum ether extract exhibiting the strongest inhibitory effect (P<0.05). The petroleum ether phase was further separated into four fractions, with the Fr.1, Fr.2 and Fr.3 fractions had stronger anti-inflammatory effects, though the Fr.1 and Fr.2 fractions contained potential toxic components. Therefore, the Fr.3 fraction was selected for further separation. The Fr.3 fraction was separated through a silica gel column to obtain three fractions. The Fr.3.1 subfraction exhibited the strongest inhibitory effect against the NO secretion (11.80 μmol/L). The Fr.3.1 subfraction was further purified by the preparative liquid chromatography and GC-MS analysis, and the main components of the Fr.3.1 subfraction were identified as stearic acid (47.09%), di(2-ethylhexyl)phthalate (13.21%) and other components. This study established a method for separating and purifying anti-inflammatory substances from purslane, and provides a theoretical reference for the development and utilization of purslane.