Unravelling the Antioxidant Potential of Cordyceps militaris andCordycepin: A Comprehensive Assay Evaluation

Introduction: Cordyceps militaris is prized for its bioactive nucleoside cordycepin, but comparative antioxidant data for the entire-fungus preparations and the pure material are limited. Oxidative stress underlies numerous chronic conditions, underscoring the need for safe, natural antiglycative and antioxidant agents. In the current work, the antioxidant potential of the methanolic extract of C. militaris [CME] was compared with that of the principal nucleoside, cordycepin. Methods: The fruiting bodies were macerated and concentrated, and cordycepin was crystallised using sequential silica-gel chromatography. The identity of the isolate CR1 was confirmed by LC-MS/MS [m/z 252 → 136] and FTIR analyses, and its purity was confirmed by HPLC [97.15%]. Results: CME contained 21.45 ± 2.28 mg gallic-acid equivalents g⁻¹ phenolics and 10.44 ± 0.73 mg quercetin equivalents g⁻¹ flavonoids. Antioxidant capacity, as studied using DPPH, hydroxyl radical, and phosphomolybdenum tests, exhibited significantly greater IC₅₀ values for cordycepin [74.29 ± 0.855, 64.77 ± 1.227, 91.87 ± 0.579 µg mL⁻¹] compared to CME [663.62 ± 0.829, 696.25 ± 1.319, 718.64 ± 1.713 µg mL⁻¹]. Both samples exhibited strong dose-response relationships, and one-way ANOVA indicated significant treatment effects [p < 0.001]. Discussion: Analytical profiling [LCMS/MS/HPLC/FTIR] verified high purity cordycepin and, with phenolic predominance [TPC > TFC], explains the robust in vitro antioxidant effects seen for both CME and cordycepin across DPPH, hydroxyl radical, and phosphomolybdenum assays. Conclusion: The combined evidence indicates that CME and cordycepin are robust in vitro antioxidants, and the purified material exhibits near-standard activity