Liquid-liquid phase separation precedes crystallization in supercooled water-glycerol solutions

Abstract: Understanding the structural evolution of supercooled water-glycerol solutions is important for cryopreservation, yet distinguishing liquid-state transformations from ice crystallization remains challenging. Here, we investigate a deeply supercooled water-glycerol solution by X-ray photon correlation spectroscopy (XPCS) in ultra-small-angle X-ray scattering (USAXS) geometry, combined with wide-angle X-ray scattering (WAXS). This combination simultaneously captures the structural and dynamical evolution of the supercooled liquid upon quenching to cryogenic temperatures (172 K). We observe discontinuous changes in the liquid structure on molecular length scales and formation of microscale domains. The dynamics slow down during this stage and exhibit hyper-diffusive, ballistic-like relaxation. This transformation precedes ice crystallization, which we identify from the emergence of ice Bragg peaks in WAXS, allowing the two processes to be temporally separated. Phase-field (Cahn-Hilliard) simulations qualitatively reproduce the experimental observations and show that a spinodal-decomposition scenario is consistent with the measured scattering evolution. These findings are consistent with a liquid-liquid phase separation scenario preceding ice crystallization and provide a route to disentangle the two processes in supercooled aqueous systems.