Freeze‐Drying of Mannitol–Trehalose–Sodium Chloride‐Based Formulations: The Impact of Annealing on Dry Layer Resistance to Mass Transfer and Cake Structure

The objective of this article was to study the mechanism by which annealing increases the primary drying time in mannitol-trehalose-sodium chloride-based formulations. The thermal events occurring during annealing and the glass transition of the frozen solutions were monitored with differential scanning calorimetry (DSC). Manometric temperature measurement was used to evaluate the dry layer resistances during primary drying. The morphologies of the freeze-dried cakes were examined by scanning electron microscopy (SEM). The degrees of crystallinity of mannitol and sodium chloride (NaCl) in freeze-dried cakes were determined by powder X-ray diffraction (XRD). DSC results indicated that annealing during freezing did not increase the glass transition temperature (Tg') significantly, but there was a distinct decrease of deltaCp at Tg' with annealing, suggesting a decrease in amorphous content. SEM revealed that most mannitol crystallized as the delta-form during annealing at -23 degrees C, and further crystallized as the alpha-form, together with NaCl crystallization, during subsequent annealing at -33 degrees C. The powder XRD results demonstrated that annealing caused crystal growth of mannitol and NaCl, and thus prevented the partial collapse observed without annealing. However, the highly crystallized mannitol blocked the pathways for water vapor escape, contributing to the increase in the dry layer resistance and thus the longer times for primary drying. Freeze-dried cakes without annealing had lower dry layer resistances because partial collapse created larger channels for water vapor escape. Therefore, two-step annealing in freezing makes mannitol-trehalose-sodium chloride-based formulations robust in freeze-drying, but annealing increases the dry layer resistances, thereby extending primary drying.