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This method determines glycerol and other polyalcohols containing three or more adjacent hydroxyl groups. The glycerol reacts with the sodium periodate in an acid solution, forming aldehydes and formic acid. The latter is a measure of the glycerol in the sample. The sodium periodate method replaces the acetin method and the dichromate method because it has been found to be more accurate and more specific for the determination of glycerol as well as being simpler and more rapid. Applicable to any glycerin solution, but particularly useful in analyses of samples containing oxidizable organic impurities and certain hydroxylated compounds which interfere with the dichromate and acetin procedures. Trimethylene glycol and other polyalcohols in which the hydroxyl groups are not adjacent do not react at room temperature. This method is not applicable to samples that contain, in addition to glycerol, polyhydric compounds with three or more adjacent hydroxyl groups, unless these compounds are removed prior to testing with the sodium periodate reagent.