HR-LCMS-Guided Phytochemical Profiling, Multi-Target Bioactivities, and In Silico Studies of Calophyllum apetalum Willd. Leaf Extracts.

This study provides a comprehensive investigation of the phytochemical composition and biological activity of Calophyllum apetalum leaf extract. Among the solvents examined, ethanol and methanol extracts possess major phytoconstituents, with methanol revealing the higher extractive value (10.45%). Several bioactive substances were found through high-resolution liquid chromatography mass spectrometry (HR-LCMS) analysis. The ethanolic extract had higher total phenolic (218.6 ± 4.16 mg gallic acid equivalent [GAE]/g) and flavonoid (256.5 ± 3.13 mg quercetin equivalent [QE]/g) contents, whereas the methanolic extract had higher terpenoid content (144.64 ± 2.03 mg LNE/g). The ethanolic extract had high antioxidant activity, including DPPH scavenging (58.47% ± 0.18%), phosphomolybdate (60.54% ± 0.15%), and metal chelation (64.91% ± 0.54%). The BSA assay demonstrated anti-inflammatory activity (59.07% ± 0.02%). At 100 µg/mL, the ethanolic extract demonstrated antibacterial action against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Bioassays using brine shrimp and hemolysis showed moderate toxicity. HeLa (54.38% ± 0.81%) and MCF-7 (51.37% ± 1.22%) cells showed reduced viability in the cytotoxicity assay by ethanolic extract. In silico docking confirmed strong binding affinities of the identified anticancer compounds to target proteins. This work presents novel HR-LCMS profiling and anticancer evaluation of C. apetalum leaf metabolites.