The adsorption of insulin at an oil-water interface was studied with fluorescence correlation spectroscopy (FCS). FCS is able to measure diffusion properties of insulin at nanomolar concentrations, making it possible to detect the very early steps in the adsorption process. Below 20 nM bulk insulin concentration, the insulin molecules adsorbed to the surface diffuse freely at all times during the experiment (a few hours). At higher concentrations, a surprisingly abrupt transition to a slow diffusion phase is observed. Based on the information about both diffusion times and molecular brightness derived from the FCS experiments, we suggest that the transition represents the formation of a fractal network. FCS may be a valuable tool in pharmaceutical formulation science, because it provides information about concentration buildup and phase changes at interfaces formed in drug delivery systems.
The results indicate that Tween 80 at concentrations above critical micelle concentration can be used to protect insulin against surface adsorption. The presence of a lag phase in the displacement at low surfactant concentration indicates that the mechanism of action for Tween 80 to reduce adsorption of insulin may be by competing for sites at the surface.
Solid insulin formulations obtained by different methods of preparation were compared with respect to chemical stability and morphology. Spray- and freeze-drying, solution enhanced dispersion by supercritical fluids (SEDS) and precipitation into starch microspheres were the methods used for preparation of solid powders. The excipients applied were melezitose, starch, and sodium taurocholate. The stability of the samples was evaluated after storage in open containers at 25 degrees C and 30% RH for 6 months. All samples were amorphous after processing and storage as detected by XRD, except for the starch microspheres which were semi-crystalline. The spray- and freeze-dried samples containing melezitose and sodium taurocholate experienced a significant water uptake during storage, resulting in changes in morphology and disappearance of Tg. However, the chemical stability of these samples did not seem to be affected by the water uptake. Changes in morphology were not observed for the SEDS powders and the starch microspheres. The chemical stability of the samples was assessed by HPLC. In general, conventional spray- and freeze drying resulted in samples with higher chemical stability compared to SEDS powders and starch microspheres. Nevertheless, the excipients applied were observed to be of major importance, and further optimization of the formulation as well as processing conditions may lead to slightly different conclusions.
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