Anionic liposomes in capillary electrophoresis: Effect of calcium on 1-palmitoyl-2-oleyl-sn-glycero-3-phosphatidylcholine / phosphatidylserine-coating in silica capillaries
2004
Hautala, Jari T. | Wiedmer, Susanne K. | Riekkola, Marja-Liisa
The effect of calcium on phospholipid coatings in fused silica capillaries used in capillary electrophoresis was studied. The anionic liposomes used for the coating consisted of 3 mM 1-palmitoyl-2-oleyl-sn-glycero-3-phosphatidylcholine and phosphatidylserine in the ratio 80/20 mol%. Coating was performed as part of the preconditioning, and the capillaries could be used for several runs without the need for liposomes in the background electrolyte solution or for liposome rinses between runs. Phospholipids could easily be flushed away by rinsing with a chloroform-methanol (2:1 v/v) mixture, which made it possible to recoat and reuse the capillaries. A calcium:phospholipid ratio of approximately 3 gave the most stable coating. The stability of the coating and success of the coating procedure were studied by measuring the electroosmotic flow and by separating uncharged steroids, which were used as model compounds. Many parameters that affect the coating, such as preconditioning (with different acids and bases), buffer, temperature during coating, and the physical structures of liposomes, were studied, with and without calcium in the liposome solution. The separation of steroids was improved and was less dependent on coating conditions when calcium was present during the coating. Capillaries optimally coated with anionic phospholipids were applied in the separation of phenols.
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