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If a 90% ethanol-water solution shall be mixed with clean water to achieve a freezing point of -20 o C (ethanol concentration 36.25% (0.3625) ) - the amount of added water can be calculated with volume balance - the amount of ethanol before mix is the same as after the mix: C V s = C m (V s + V w ) (2) where V s = volume of the ethanol - water solution (liter, gallon) C m = concentration in mix V m = volume of mix (liter, gallon) V w = volume of the added clean water (liter, gallon) Rearranging the equation to express the volume of water added to the mixture V w = (C - C m ) V s / C m (2b) Substituting with values V w = (0.9 - 0.3625) V s / 0.3625 = 1.48 V s - for every liter 90% ethanol-water solution 1.48 liter of clean water must be mixed in to achieve an ethanol concentration of 36.25 % and freezing point -20 o C .

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Wen S, Zhang T, Tan T (2004) Utilization of amino acids to enhance glutathione production in Saccharomyces cerevisiae