| Summary: | <i>Aconitum chasmanthum</i> Stapf ex Holmes, an essential and critically endangered medicinal plant from Kashmir Himalayas, was studied for its antioxidant and antifungal properties. The shade-dried powdered rhizome was extracted sequentially with hexane, ethyl acetate, and methanol. These subsequent fractions were evaluated for total phenolic content (TPC); total flavonoid content (TFC); antioxidant assays, such as 1,1-diphenyl 1-2-picryl-hydrazyl (DPPH); ferric-reducing antioxidant power (FRAP); superoxide radical scavenging (SOR); hydroxyl radical scavenging (OH) and antifungal activity using the poisoned food technique. Highest TPC (5.26 ± 0.01 mg/g) and TFC (2.92 ± 0.04 mg/g) were reported from methanolic extracts. The highest values of radical scavenging activities were also observed in methanolic extracts with IC<sub>50</sub> values of 163.71 ± 2.69 μg/mL in DPPH, 173.69 ± 4.91 μg/mL in SOR and 159.64 ± 2.43 μg/mL in OH. The chemical profile of ethyl acetate extract was tested using HR-LCMS. Methanolic extracts also showed a promising inhibition against <i>Aspergillus niger</i> (66.18 ± 1.03), <i>Aspergillus flavus</i> (78.91 ± 1.19) and <i>Penicillium notatum</i> (83.14 ± 0.97) at a 15% culture filtrate concentration with minimum inhibitory concentration (MIC) values of 230 μg/mL, 200 μg/mL and 190 μg/mL, respectively. Overall, the methanolic fractions showed significant biological potential, and its pure isolates might be used to construct a potential new medicinal source.
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