Glycerin is used in different concentrations according to the intended use such as an antimicrobial preservative, emollient, aqueous and non-aqueous gel vehicle, humectant, and plasticizer in tablet film coating and sweetening agent in alcoholic elixirs. A precise, accurate, and reversed-phase high-performance liquid chromatographic (HPLC) method was developed for the determination of glycerin in different pharmaceutical preparations. Analysis was performed on Microsorb-MV 100-5 NH2 (150 x 4.6 mm) or equivalent column with an isocratic mobile phase consisting of hexane buffer 0.005M (pH 3), Acetonitrile, and Methanol (12:78:10 v/v/v) at a flow rate of 1.0 mL/min and ambient temperature using photodiode array (PDA) detector at 202 nm. The retention time of the drug was found to be 3.011 ± 0.007 min. The injection volume was 20 µL. The mobile phase was used as a diluent during the standard and test sample preparation. The developed method was found to be linear over the range of 3.75-22.5 mg/mL for glycerin. The currently developed method can be easily performed in quality control laboratories for simultaneous determination of glycerin, where no expensive tools are available.