Specific protection mechanism of oligosaccharides on liposomes during freeze-drying

Liposomes have been received much attention during the past decades as bioactive compounds carriers in food field. However, the application of liposomes is extremely limited by the structural instability during processing such as freeze-drying. In addition, the protection mechanism of lyoprotectant for liposomes during freeze-drying remains controversial. In this study, lactose, fructooligosaccharide, inulin and sucrose were used as lyoprotectants for liposomes and the physicochemical properties, structural stability and freeze-drying protection mechanism were explored. The addition of oligosaccharides could significantly suppress the changes in size and zeta potential, and the amorphous state of liposomes was negligible changed from XRD. The T_g of the four oligosaccharides, especially for sucrose (69.50 °C) and lactose (95.67 °C), revealed the freeze-dried liposomes had formed vitrification matrix, which could prevent liposomes from fusion via increasing the viscosity and reducing membrane mobility. The decrease in T_m of sucrose (147.67 °C) and lactose (181.67 °C), and the changes in functional group of phospholipid and hygroscopic capacity of lyophilized liposomes indicated oligosaccharides replaced water molecules to interact with phospholipids by hydrogen bonds. It can be concluded that the protection mechanism of sucrose and lactose as lyoprotectant was attributed to the combination of vitrification theory and water replacement hypothesis, while the water replacement hypothesis was dominated by fructooligosaccharide and inulin.