Simple coating of capillaries with anionic liposomes in capillary electrophoresis

被引:70
作者
Hautala, JT
Lindén, MV
Wiedmer, SK
Ryhänen, SJ
Säily, MJ
Kinnunen, PKJ
Riekkola, ML
机构
[1] Univ Helsinki, Dept Chem, Analyt Chem Lab, FIN-00014 Helsinki, Finland
[2] Univ Helsinki, Biomedicum, Inst Biomed, Helsinki Biophys & Biomembrane Grp, FIN-00014 Helsinki, Finland
基金
芬兰科学院;
关键词
coated capillaries; liposomes; electroosmotic flow; background electrolyte composition; phospholipids; hydroxyethylpiperazine-ethanesulfonic acid;
D O I
10.1016/S0021-9673(03)00570-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new and relatively simple method was developed for coating of capillaries in electrophoresis with liposomes. The liposomes, with a diameter of about 100 run, are large unilamellar vesicles prepared by extrusion. The liposomes contained 1-palmitoyl-2-oleyl-sn-glycero-3-phosphatidylcholine (POPC) or POPC with different proportions of bovine brain phosphatidylserine (PS) and cholesterol. They formed a bilayer structure on the silica surface enabling the separation of neutral compounds. The effectiveness of the coating in separation was evaluated with use of uncharged steroids as model compounds. The coating was also studied by measuring the electroosmotic flow. The best results, taking into consideration both separation and stability, were achieved with anionic 80:20 mol% POPC/PS liposomes. In addition, the effect of coating conditions on the results was investigated. Among the buffers studied [N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), phosphate, tris(hydroxymethyl)aminomethane (Tris) and N-tris(hydroxymethyl)methylglycine (Tricine)], HEPES seemed to have a significant effect on the success of the coating. Successful separation of steroids was achieved only when HEPES buffer was used in the coating procedure and in the background electrolyte solution for the separation. With all other buffers the peaks of the model compounds overlapped. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:81 / 90
页数:10
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