Artemisia rutifolia (Asteraceae) had been used in traditional medicines for the treatment of different ailments. In the current study, an effort was made to explore the phenolic composition, antioxidant, and antimicrobial activities of different solvent extracts obtained from A. rutifolia leaves. The reverse-phase high-performance liquid chromatographic (RP-HPLC) analysis revealed the higher extent of polyphenolic compounds (i.e., gallic acid, caffeic acid, chlorogenic acid, syringic acid, sinapic acid, p-coumaric acid, m-coumaric acid, ferulic acid, vanillic acid, myricetin, and quercetin) in methanol extract. Methanol extract consistently showed the highest total phenolic contents (98 ± 2 µg GAE/mg of plant extract), total flavonoid contents (28 ± 0.0 µg QE/mg of plant extract), antimicrobial activity, free radical (DPPH) scavenging (IC 50 = 39 µg/mL) activity, and reducing power (18.3 ± 0.2 mg GAE/g of plant extract) followed by those of chloroform and hexane extracts, respectively. The current study concluded that extracts of A. rutifolia are novel natural source of antioxidative and antimicrobial agents for the treatment of oxidative stress-related disorders and microbial infections.