1. The amount of free unfrozen water, i.e. water acting as normal solvent, in frog's muscle at temperatures below the initial freezing-point has been calculated from the vapour pressure isotherm of the muscle.
2. Significant amounts of free water are present at –20°C. The total amount of unfrozen water at –20°C. cannot, therefore, be taken as a measure of the bound water in muscle.
3. The calculated values of free water, when compared with experimentally determined values of total unfrozen water, indicate that the amount of bound water in muscle at various temperatures is small.
4. A temperature considerably below –20°C., roughly between –40° and –60°C., is required to freeze completely the free water in muscle.