Anti-solvent effect in the production of lysozyme nanoparticles by supercritical fluid-assisted atomization processes

Particles of lysozyme in the range of 0.1-5 μm were generated by high pressure CO₂ or N₂ (at pressures between 8 MPa and 25 MPa) from aqueous ethanol solutions using an atomization process similar to the supercritical assisted atomization technology. Perfect nanosized spheres of lysozyme were produced using both supercritical fluids. However, while N₂ assisted atomization-produced spheres at all experimental conditions reported here, supercritical CO₂ assisted atomization produced particles of two distinct morphologies depending on the pre-mixing conditions. This work shows that CO₂ assisted atomization produces particles by two different mechanisms depending on the mixture pre-expansion phase equilibria conditions: anti-solvent crystallization and spray drying crystallization. Depending on the governing precipitation mechanism (anti-solvent or spray drying), fibers or spherical particles were obtained with CO₂. Lysozyme activity was severely affected by pure anti-solvent processing, while N₂ processed lysozyme conserved mostly its activity.