Impact of Heat Treatment on the Physical Properties of Noncrystalline Multisolute Systems Concentrated in Frozen Aqueous Solutions

The purpose of this study was to elucidate the effect of heat treatment on the miscibility of multiple concentrated solutes that mimic biopharmaceutical formulations in frozen solutions. The first heating thermal analysis of frozen solutions containing either a low‐molecular‐weight saccharide (e.g., sucrose, trehalose, and glucose) or a polymer (e.g., polyvinylpyrrolidone and dextran) and their mixtures from −70°C showed a single transition at glass transition temperature of maximally freeze‐concentrated solution (Tg′) that indicated mixing of the freeze‐concentrated multiple solutes. The heat treatment of single‐solute and various polymer‐rich mixture frozen solutions at temperatures far above their Tg′ induced additional ice crystallization that shifted the transitions upward in the following scan. Contrarily, the heat treatment of frozen disaccharide‐rich solutions induced two‐step heat flow changes (Tg′ splitting) that suggested separation of the solutes into multiple concentrated noncrystalline phases, different in the solute compositions. The extent of the Tg′ splitting depended on the heat treatment temperature and time. Two‐step glass transition was observed in some sucrose and dextran mixture solids, lyophilized after the heat treatment. Increasing mobility of solute molecules during the heat treatment should allow spatial reordering of some concentrated solute mixtures into thermodynamically favorable multiple phases.