PROCEDURE FOR PREPARATION OF LIPID VESICLES (LIPOSOMES) USING THE COACERVATION (PHASE-SEPARATION) TECHNIQUE

The preparation of lipid vesicles using simple coacervation is described in detail. The optimal coacervation conditions for the formation of lipid vesicles by the triangular phase diagram of the lipid-alcohol-water system were examined. Four different alcohols (methanol, ethanol, n-propanol, and 2-propanol) were used as a lipid solvent, and deionized, distilled water was used as a poor solvent for the lipids. The lamellarity and size homogeneity of the resulting lipid vesicles were observed by freeze-fracture electron microscopy and scanning electron microscopy, respectively, using a specific fixation technique with malachite green. The majority of vesicles formed by methanol as a lipid solvent appeared to be large and unilamellar, ranging from 100 to 1000 nm in diameter. However, when the vesicles were prepared using ethanol, concentric lamellae of multilamellar vesicles were recognizable. On the other hand, the use of propanol resulted in lipid vesicles that were homogeneous and unilamellar.