This work is focused on the study of the effect of cholesterol on the properties of vesicular membranes of ionic amphiphilic pairs at different temperatures. The hexadecyltrimethylammonium-dodecyl sulfate ionic amphiphilic pair system with the addition of 10 mol % dioctadecyldimethylammonium chloride was chosen for a detailed study of vesicle properties. A large range of cholesterol concentrations (0–73 mol %) in the temperature range 10–80 °C was studied. Under these conditions, the size distribution, the membrane fluidity, and the surface layer were monitored together with the change in the mobility of water in the surface layer. Obtained quantities were correlated with each other and combined into appropriate graphs. It was found that in stable systems that meet the condition of unimodal size distribution with a PDI value lower than 0.3, temperature has virtually no effect on the size of vesicular systems. On the contrary, when studying the hydration and fluidity of the membrane, significant changes in these parameters were found, which, however, do not affect the short-term stability of these vesicular systems. The presented results thus indicate the possibility of adjusting the composition of the vesicular system in terms of fluidity and membrane hydration while maintaining short-term stability and size distribution.

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