Glutathione (GSH) is an intracellular antioxidant that plays vital role in maintaining redox homeostasis. As a result, GSH serves as an important biomarker for understanding various physiological disorders, including cancer as well as multiple cardiovascular and liver-related diseases. Therefore, the sensitive detection of GSH is crucial in fields such as drug delivery and biological responses. In recent years, the use of transition metal oxides for GSH monitoring has gained considerable attention. In this work, we investigate the potential of hydrothermally synthesized α-MnO2 nanostructures for GSH sensing. X-ray diffraction, FTIR, and Raman spectroscopy were employed to gain insights into the structural characteristics of the material. FESEM images revealed that the MnO2 nanostructures exhibit an elongated nanowire morphology. Optical analysis using UV-visible spectroscopy confirmed the semiconducting nature of the sample, with a bandgap of 2.75 eV. The MnO2 solution was prepared in citrate buffer under acidic pH conditions and upon the addition of various concentrations of GSH, a noticeable change in the color intensity of the MnO2-citrate buffer solution was observed. This shift, as evident from the UV-visible absorption spectra, enables the colorimetric detection of GSH. These findings clearly establish α-MnO2 nanorods as a highly effective and promising material for the sensitive detection of GSH.