Glycerol is a polyol hydrophilic compound which is used in pharmaceutical formulations. It is also a byproduct of biodisel production through transesterification. Glycerol has three hydroxyl groups, making it a very polar compound. Method using periodate/acetylacteone derivatization was developed using UV. Method is fast and shows good linearity within explored range of concentrations. This general HPLC approach can be used for precise quantitation of glycerol.