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Artemisia herba-alba L. (A. herba-alba), growing in the Mediterranean regions, contains various phytochemical compounds with exceptional pharmacological and chemical properties. The present study aims to characterize chemical and biological activities of A. herba-alba's methanolic extract according to total protein, total phenols, antioxidant, anti-inflammatory, and anti-hemolytic activities. The study also evaluates the extract's anti-proliferative activity of diverse cancer cell lines in vitro. Liquid chromatography-mass spectrometry (LC-MS) was used to conduct phytochemical analysis of A. herba-alba's extract. The total phenolic amount, total proteins, anti-inflammatory, and anti-hemolytic activity was evaluated in vitro alongside antioxidant activity by various spectrophotometric methods. LC-MS analysis of crude extract reveals that kaempferol, quercetin, geranyl acetate, dihydrolinalool, furanone, vetivenic acid, limonene, eugenol, p-cymene, and naringin represent the major composition (81.5%). A. herba-alba's extract showed 72.07 % antioxidant activity through DPPH radical scavenging with IC50 value 0.67 μg/ml. Furthermore, the extract from A. herba-alba had a 68.71% antihemolytic impact and a 97.55% anti-inflammatory effect. These different bioactivities were linked and correlated to the total phenolic content in the extract (1.41 mg/ml). The correlation between phenolic contents and activity of DPPH scavenging, anti-inflammatory, and anti-hemolytic was expressed with Pearson correlation coefficient and found to be a strong correlation of the extract with R² close to 0.90. Furthermore, strong cytotoxic activity was shown by the extract against tested cancer cell lines. The IC50 values for lung cancer HCC95 was 7.0 μg/mL, breast cancer MDA-MB-231 was 6.9 μg/mL, prostate DU-145 was 6.8 μg/mL, and breast cancer 600MPE was 6.7 μg/mL. According to the research, the methanolic extract of A. herba-alba plant contains numerous phytochemicals, primarily phenolic compounds. The extract exhibits antioxidant, anti-inflammatory, and anti-hemolytic activities, as well as strong cytotoxic activity. [ABSTRACT FROM AUTHOR]