Abstract
Purpose: Ocimum basilicum L., a medicinal herb from the Lamiaceae family, has been traditionally used to treat various diseases. Plant extracts are valued in therapeutic medicine for their rich bioactive compounds. Selecting the appropriate extraction method and solvent is crucial for obtaining superior-grade plant extracts. This research is aimed at characterizing the phytochemical profiles and potential biological activities of hydromethanolic (HME) and n-hexane (HE) extracts derived from Ocimum basilicum L. leaves using the soxhlation extraction method. Methods: Extracts were subjected to physiochemical analysis (FTIR, GC-MS, and TLC/HPTLC), quantitative analysis (total phenolic, flavonoid, and tannin content), antioxidant activity (DPPH, ABTS, NO, and SO assays), toxicity (brine shrimp and hemolysis), and anticancer activity (MTT). Results: FTIR analysis revealed major functional groups in HME and fewer in HE. GC-MS analysis identified various compounds in HME, including estragole, beta-farnesene, alpha-bergamotene, alpha-copaene, eicosapentaenoic acid, and neophytadiene, while HE contained estragole, 1-heptatriacotanol, alpha-bergamotene, and tau-cadinol. TLC/HPTLC analysis reflects that both extracts contain complex mixtures of bioactive compounds. HME exhibited significantly higher levels of phenolics (122.72 ± 4.02 mg GAE/g), flavonoids (42.47 ± 0.59 mg QE/g), and tannins (30.91 ± 0.31 mg TAE/g) compared to HE (75.94 ± 0.93, 26.65 ± 1.17, and 1.99 ± 0.07 mg/g, respectively). In addition, HME exhibited significantly stronger antioxidant activity, as evidenced by lower IC50 values across all assays (DPPH, ABTS, NO, and SO) compared to HE. Both HME and HE exhibited notable cytotoxic activity against HCT 116, MCF-7, and HeLa cancer cell lines in MTT assays. HE showed significantly greater cytotoxicity than HME at both 24 and 48 h. However, HME showed no toxicity in the brine shrimp lethality assay even at 2000 μg/mL and exhibited no hemolytic activity, highlighting its strong biocompatibility. Conversely, HE presented mild toxicity (LC50 = 992 ± 88.34 μg/mL) and significant hemolytic effects, indicating potential risks to erythrocytes, limiting its therapeutic use unless mitigated. Conclusion: Integrating metabolomics and pharmacognostic studies provided a comprehensive understanding of Ocimum basilicum L. extracts, highlighting their quality, safety, and medicinal potential.