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Background: Dexamethasone (DEXA) is a potent synthetic corticosteroid derived from the cyclopentanoperhydrophenanthrene nucleus known for its anti-inflammatory and immunosuppressive activities. Due to its therapeutic effects, several analytical methods have been used for its quantitative determination and for physicochemical characterization, as well as for evaluation of pharmacological and toxicological properties. Objective: This review was aimed to describe the principles and methods commonly used to identify and quantify DEXA in drug delivery systems and biological samples. The methods herein discussed are high performance liquid chromatography, nuclear magnetic resonance, x-ray diffraction, Fourier-transform infrared spectroscopy, differential scanning calorimetry, ultraviolet-visible spectrophotometry and thin layer chromatography. Conclusion: This review provided a wide variety of analytical methods that can be used for the quantification and identification of drugs, providing scientists with great support during the development of scientific research, as well as ensuring the quality of the manufacturing processes as well as of the resulting products. Therefore, the use of such analytical methods has become critical throughout the process of developing pharmaceutical formulation containing DEXA.