IN VITRO INVESTIGATION OF ANTIOXIDANT AND ANTICOAGULANT PROPERTIES OF Calocybe indica (MILKY MUSHROOM)

Abstract

The growing demand for natural therapeutic compounds has intensified the search for bioactive molecules with diverse therapeutic properties, including antioxidant and anticoagulant activities. Calocybe indica (milky mushroom), an edible mushroom widely cultivated on a large scale in South Asia, is known for its richness in phytochemicals. However, research on C. indica is relatively less, particularly regarding its bioactivity and the majority of the studies were carried out on its nutrient content. Therefore, this study was conducted to evaluate and compare the antioxidant and anticoagulant activities of aqueous, methanol, dichloromethane (DCM), and hexane extracts of C. indica fruiting bodies. Authenticated fruiting bodies of C. indica were freeze-dried, cold macerated, and concentrated. Total phenolic content (TPC) was determined using the Folin–Ciocalteu method, while antioxidant activity was measured using the 2, 2-diphenyl-1 picrylhydrazyl (DPPH) radical scavenging assay. Among the extracts, the methanol and aqueous extracts exhibited the highest antioxidant activity, with the lowest IC₅₀ values of 561.7 mg/mL and 549.8 mg/mL, respectively, indicating strong free radical scavenging potential. These were followed by the hexane (788.3 mg/mL) and DCM (961.6 mg/mL) extracts. A similar trend was observed in TPC values, further confirming these activities, with the aqueous extract showing the highest TPC (176.96 mg GAE/g), demonstrating a strong correlation between phenolic content and antioxidant activity. Anticoagulant activity was evaluated by measuring the prolongation of prothrombin time (PT). IBM-SPSS version 26 was used for statistical analysis. The bar chart analysis indicated that all four extracts, at varying concentrations, caused a significant increase in PT values compared to the control, confirming anticoagulant potential. All extracts contributed to prolonged coagulation time, highlighting the potential of C. indica as a natural anticoagulant. This study highlights the promising antioxidant and anticoagulant potential of C. indica, supported by its high phenolic content and ability to prolong prothrombin time. These findings are consistent with previous studies that report the therapeutic effects of mushroom-derived compounds such as polysaccharides and phenolics. Further research is needed to isolate the active constituents and confirm their efficacy and safety through in vivo studies. C. indica shows strong potential for development as a natural therapeutic agent against oxidative stress and thrombotic disorders