超临界CO_2提纯肺表面活性物质及其产物的化学成分和活性功能测定

Purifying Pulmonary Surfactant by Supercritical Carbon Dioxide Extraction and Determination of the Chemical Composition and the Surface Activity of the Production

目的建立超临界CO2流体萃取技术提纯肺表面活性物质的新方法,并对萃取所得产物进行化学成分分析和体外活性测定。方法运用超临界CO2流体萃取技术去除肺表面活性物质粗品中的中性脂质等低极性成分,以提纯肺表面活性物质。提纯获得的产物经薄层色谱和高效液相色谱对其化学成分进行分析,产物的体外活性功能用动态膜压记录仪测定。结果经测定,所得产物中总磷脂含量高达(91.33±1.58)%;其中含有6种磷脂成分,最主要的活性物质-磷脂酰胆碱的含量达(81.42±2.35)%;动态膜压记录仪测其生物学活性,可将最小表面张力降到10mN/m以下。结论该提纯方法简便、高效、有机溶剂污染少,且提纯过程不影响PS产物的活性功能。

Aim To develope a supercritical fluid extraction method for purifying natural pulmonary surfactant. Then to determine the chemical composition and the surface activity of the product. Methods The present research relates to a process for the purification of pulmonary surfactant by means of extraction and separation of undesired components consisting of lipidic substances of a low polar power. They were extracted and separated by SFE - CO2. The chemical composition of the product was analyzed by thin - layer chromatography and high performance liquid chromatograph. The surface activity of the product was determined by film balance. Results Phospholipids were found to be the main component, composing （91.33 ± 1.58） %. It was found that the product contained six kinds of phospholipids,among which phosphatidylcholine （PC） was the chief substance （81.42 ± 2.35）%. Surface activity of the product reduced minimal surface tension to less than 10mN/m. Conclution The new method for purifying natural pulmonary surfactant by SFE - CO2 can cause drastic reduction of organic solvents, being convenient to use and effective to purify. Furthermore, the novel purification procedure does not affect the biological properties in any way.