*2.5. Preparation of Spirulina Hydrolysate (HS)-Loaded Liposomes*

HS-encapsulated nanoliposomes were prepared using the thin layer hydration method as described by Mohammadi et al. [7] with slight modifications. For this procedure, 1.2 % (*w*/*v*) Phospholipon 90 G (soybean lecithin of ~90% phosphatidylcholine; Lipoid GmbH, Ludwigshafen, Germany) and 2 stabilizing agents (cholesterol or 0.15% (*w*/*v*) γ-oryzanol) were dissolved in 15 mL of 96% ethanol and stirred on a hotplate at 50 ◦C for complete solubilization. Subsequently, the solvent was evaporated using a rotary evaporator (Heidolph, Germany) at 50 ◦C until a thin film was formed in the roundbottomed flasks. The resulting lipid films were hydrated with 15 mL of DW containing HS at 0.3% (*w*/*v*) with continuous agitation on a rotary evaporator at 55 ◦C, followed by sonication using a sonication probe (130 W, 20 kHz; Vibra-Cell Sonics & Materials, Newtown, CT, USA) at 80% sonication strength for 10 min. During sonication, the sample was placed into an ice bath to avoid overheating of dispersion. To prepare the empty liposomes, the same method was applied, except HS was excluded in the hydration step and the thin layer was hydrated only with DW.
