Melasma is an abnormal acquired skin hyperpigmentation disorder, typically on the face, of unknown origin. It is considered a single disease and very little has been found regarding its pathogenesis. Hydroquinone, an aromatic organic molecule, has been considered as the gold standard substance for dermatological melasma treatment. This substance at high concentrations being absorbed by the skin may produce counterproductive disorders, such as blue or brownish-blue colored skin. In recent years, optical techniques based on the interaction of light with biological samples have become innovative methods for medical applications. We used Raman spectroscopy and surface-enhanced Raman spectroscopy (SERS), to evaluate hydroquinone cation radicals in vitro at relatively low concentrations, laying the foundation for future biomedical applications. We present the experimental and simulated Raman signal in the presence of hydroquinone at concentrations as low as 0.1 M, as well as the experimental SERS signal assisted by gold nanorods obtained for the same molecule, which presents an electromagnetic enhancement factor of ∼10⁴.