This study explores the phytochemical composition and biological activity of petroleum ether extract (PEEM) derived from the mace of Endocomia macrocoma. Methods employed include qualitative phytochemical screening, HPTLC, GC-MS, LC-MS, as well as free radical scavenging and antimicrobial assays. Preliminary screening revealed the presence of terpenoids, glycosides, alkaloids and tannins. Antioxidant activity was confirmed via HPTLC-DPPH and ABTS assays showing yielding IC₅₀ values of 143.68 ± 2.82 μg/mL and 105.28 ± 1.36 μg/mL, respectively. As the first comprehensive analysis of E. macrocoma mace, this study highlights its potential as a natural source of antioxidants and antimicrobial agents, supporting traditional medicinal use and offering prospects for pharmaceutical development.