Analysis of the Particle Size Distribution and Internal Volume of Liposomal Preparations

In this work we studied the particle size distribution of three liposomal preparations by quasi-elastic light scattering spectroscopy. Sized unilamellar vesicles of small diameter (s-SUV) were prepared by ultrasonication and subsequent centrifugation followed by extrusion through polycarbonate membranes of 0.2-µm pore size. Large unilamellar vesicles were obtained by reversed-phase evaporation (REV) and extrusion through polycarbonate filters with or without preliminary freezing-thawing cycles (VET, and VETM, respectively). After preparation, REV were sized to small diameter REV (s-REV) by extrusion through 0.4- and 0.2-µm polycarbonate membranes.According to the results, the s-SUV preparations were made up of two subpopulations, the major of which consisted of vesicles that were 26nm in mean diameter and accounted for 95% of the overall s-SUV population. The s-REV dispersions always resolved into two populations centered at 120 and 380nm, the relative proportions of which depended on the pore size of the filters used. VET structures were composed of a single population of vesicles that were -100nm in mean diameter. Cholesterol inclusion into the bilayer composition extended the distribution without altering its mean value. On the other hand, the internal volumes calculated from mean diameters or assuming a Gaussian distribution were inconsistent with experimental data obtained by usual techniques.