Formation of microparticulate protein powder using a supercritical fluid antisolvent

Gas antisolvent (GAS) expansion of dimethylsulfoxide (DMSO) and N,N‐dimethylformamide (DMFA) solutions with supercritical carbon dioxide was used to produce biologically active powders of insulin. Powders with 90% of the particles smaller than 4 μm and 10% smaller than 1 μm were obtained under all conditions tested when the process was operated continuously, with small liquid droplets sprayed into a flowing supercritical continuum. Slow pressurization of the stagnant protein solution resulted in larger particles. In vivo tests on rats revealed no differences between the biological activity of processed and unprocessed insulin, GAS processing of organic solution appears to be a reliable and effective method for the production of dry, biologically active microparticulate powders of peptides and proteins. © 1993 John Wiley & Sons, Inc.