A metastable state of high surface activity produced by sonication of phospholipids

Sonication of phosphatidylcholine dispersions generates a metastable high energy assembly of molecules, the existence of which is revealed by its conspicuous surface activity. Freshly sonicated liposome dispersions release molecules to the air/water interface at rates sufficient to produce a close-packed monolayer within minutes. In contrast, monolayers at the surface of multilamellar and extruded vesicles take hours to form. The highly surface active species appears within the first few minutes of sonication, long before a major reduction in turbidity occurs, and accumulates over the course of a few hours of sonication. It disappears upon exhaustive sonication, extrusion, addition of extruded vesicles, or, more slowly, simply on standing. Tests for extraneous substances in the lipids before as well as after sonication revealed amounts of degradation products too small to represent the observed surfactant. Direct evidence that the metastable aggregate releases intact phospholipids was provided by a novel procedure to characterize monolayer composition by comparing surface tension with surface potential, both as a function of surface density. Centrifugation and gel filtration chromatography indicate that the surface activity is associated with a particle of diameter larger than a lysophosphatidylcholine micelle but not larger than limit sonicated vesicles. The metastable material appears to be lipid molecules in other than the normal stable vesicular state, perhaps an incompletely closed vesicle, one in which the inner and outer monolayers have not equilibrated, or possibly a micellar form.