通过高效凝胶排阻色谱法研究囊泡通透性的方法学。

Methodology for vesicle permeability study by high-performance gel exclusion chromatography.

作者信息

Andrieux K, Lesieur S, Ollivon M, Grabielle-Madelmont C

机构信息

Equipe Physicochimie des Systèmes Polyphasés, Université Paris-Sud, URA CNRS 1218, Châtenay-Malabry, France.

出版信息

J Chromatogr B Biomed Sci Appl. 1998 Feb 27;706(1):141-7. doi: 10.1016/s0378-4347(97)00517-3.

DOI:10.1016/s0378-4347(97)00517-3

PMID:9544816

Abstract

A methodology based on high-performance gel exclusion chromatography (HPLC-GEC) has been developed to perform permeability studies of vesicles. Encapsulation of two marker isothiocyanate fluorescein (FITC) dextrans of 4400 and 40,500 molecular mass was used as a model system. Combination of two TSK-PW columns, one efficient in vesicle sizing (G6000 PW), the other in that of dextrans (G4000 PW), was required to achieve complete particle separation and to remove entirely the unentrapped dextran after encapsulation into vesicles. Coupling fluorescence and light scattering detection allowed to control the efficiency of the separation, to quantify the vesicle leakage and to follow both the integrity of the vesicles and changes in their size. This methodology can be applied to other fields such as encapsulation of water soluble compounds and drug delivery systems.

摘要

已经开发出一种基于高效凝胶排阻色谱法（HPLC - GEC）的方法来进行囊泡的通透性研究。使用包封两种分子量分别为4400和40500的异硫氰酸荧光素（FITC）标记葡聚糖作为模型系统。需要将两根TSK - PW柱组合使用，一根用于高效测定囊泡大小（G6000 PW），另一根用于测定葡聚糖大小（G4000 PW），以实现完全的颗粒分离，并在将葡聚糖包封到囊泡后完全去除未包封的葡聚糖。荧光检测与光散射检测相结合，可以控制分离效率，量化囊泡泄漏，并跟踪囊泡的完整性及其大小变化。这种方法可应用于其他领域，如水溶性化合物的包封和药物递送系统。