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Updated: Jul 25, 2025

A Microfluidic Approach for the Study of Ice and Clathrate Hydrate Crystallization
Published on: August 18, 2022
Probing the Mechanism of Action of Small-Molecule Ice Recrystallization Inhibitors Using Proton Nuclear Magnetic
Leah E McMunn1, Adrian S D'Costa1, Nicolas Bordenave1,2,3
1Department of Chemistry and Biomolecular Sciences, Faculty of Science, University of Ottawa, STEM Complex, Room 342, 150 Louis-Pasteur Pvt, Ottawa, ON Canada, K1N 6N5.
Abstract:
N-2-Fluorophenyl-d-gluconamide (2FA) improves the recovery and function of cryopreserved biological materials by inhibiting ice recrystallization. However, as for many small-molecule ice recrystallization inhibitors, the mechanism of action of 2FA is not well-understood. In this study, the IC50 of 2FA for ice recrystallization was determined to be 3.5 mM (95% CI [3.41-3.52]). 1H transverse and longitudinal relaxations were then characterized by NMR at 2FA concentrations from 0 to 10 mM and at temperatures between -15 °C and +30 °C. Corresponding activation energy of water molecule motion (E) was calculated, showing that at each concentration 2FA did not affect E in the solid state, whereas in the liquid state E was significantly higher with 2FA than for pure water. Therefore, 2FA is excluded from the ice lattice upon freezing and concentrated in the interstitial liquid phase. This restricts the migration of water molecules between ice crystals via the liquid phase, inhibiting ice recrystallization.
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