Ice Nucleation in Emulsified Aqueous Solutions of Antifreeze Protein Type III and Poly(vinyl alcohol)

Antifreeze protein (AFP) III and poly(vinyl alcohol) (PVA) are known as anti-ice nucleating agents (anti-INAs), which inhibit heterogeneous ice nucleation. However, the effectiveness of these anti-INAs in inhibiting ice nucleation in water-in-oil (W/O) emulsions, in which homogeneous ice nucleation can be experimentally simulated, is unclear. In this study, the ice nucleation temperature in emulsified solutions of AFP III, PVA, and other nonanti-INA polymers was measured, and then the nucleation rate was analyzed based on classical nucleation theory. Results showed that ice nucleation was surface-initiated and, except for PVA solutions, probably caused heterogeneously by the emulsifier, SPAN 65, at the droplet surfaces. In this nucleation mode, AFP III had no significant effect on the ice nucleation rate. In contrast, PVA exhibited ice-nucleating activity only at the droplet surfaces, suggesting that the nucleation is due to the interaction between PVA and SPAN 65.