Preparation and characterization of microparticles containing peptide produced by a novel process: spray freezing into liquid

Abstract

The objective of this study is to evaluate excipient type on the physicochemical properties of insulin microparticles produced by spray freezing into liquid (SFL). A novel process was developed to produce microparticles containing bioactive peptides and proteins. The microparticles were formed by atomization of an aqueous feed solution containing insulin beneath the surface of a cryogenic liquid (e.g. liquid nitrogen). In this study, bovine insulin was dissolved in deionized water alone or with tyloxapol, lactose or trehalose. The aqueous solution was sprayed directly into liquid nitrogen through a polyetheretherketone capillary nozzle under high pressure to form frozen microparticles. Lyophilization was used to sublime the ice. The SFL insulin powder was characterized by different techniques, including X-ray diffraction, reverse-phase high pressure liquid chromatography, size exclusion chromatography, scanning electron microscopy (SEM), particle size distribution and surface area. The mean diameter of the insulin microparticles was 5–7 μm. SEM revealed that the microparticles were highly porous, and the morphology of the microparticles was influenced by the excipient type. The total surface area of the insulin microparticles ranged from 20 to 40 m 2/g, and the magnitude depended on the specific composition and total solids content of the aqueous feed solution. X-ray diffraction results indicated lack of crystallinity. No change in the level of the degradation product, A-21 desamido insulin, was found in the SFL insulin samples processed alone or with trehalose or tyloxapol. Similarly, no change in formation of high molecular weight transformation products (e.g. covalent insulin dimer) was detected in the samples processed with excipients. The results demonstrated that SFL is a feasible technique for forming porous microparticles containing insulin. The physicochemical properties of insulin were preserved by the SFL technique.