Investigation of PEG Crystallization in Frozen PEG-Sucrose-Water Solutions. I. Characterization of the Nonequilibrium Behavior during Freeze-Thawing

Our objective was to characterize the nonequilibrium thermal behavior of frozen aqueous solutions containing PEG and sucrose. Aqueous solutions of (i) sucrose (10%, w/v) with different concentrations of PEG (1–20%, w/v), and (ii) PEG (10%, w/v) with different concentrations of sucrose (2–20%, w/v), were cooled to −70°C at 5°C/min and heated to 25°C at 2°C/min in a differential scanning calorimeter. Annealing was performed at temperatures ranging from −50 to −20°C for 2 or 6 h. Similar experiments were also performed in the low-temperature stage of a powder X-ray diffractometer. A limited number of additional DSC experiments were performed wherein the samples were cooled to −100°C. In unannealed systems with a fixed sucrose concentration (10%, w/v), the T′g decreased from −35 to −48°C when PEG concentration was increased from 1% to 20% (w/v). On annealing at −25°C, PEG crystallized. This was evident from the increase in T′g and the appearance of a secondary melting endotherm in the DSC. Low- temperature XRD provided direct evidence of PEG crystallization. Annealing at temperatures ≤−40°C did not result in crystallization and a devitrification event was observed above the T′g. In unannealed systems with a fixed PEG concentration (10%, w/v), the T′g increased from −50 to −40°C when sucrose concentration was increased from 5% to 50%, w/v. As the annealing time increased (at −25°C), the T′g approached that of a sucrose-water system, reflecting progressive PEG crystallization. A second glass transition at ~−65°C was evident in unannealed systems [10%, w/v sucrose and 10 (or 20%), w/v PEG] cooled to −100°C. Investigation of the nonequilibrium behavior of frozen PEG-sucrose-water ternary system revealed phase separation in the freeze-concentrate. Annealing facilitated PEG crystallization.