Liquid Chromatography–High‐Resolution Mass Spectroscopy‐Based Metabolomics for Identification Cytotoxic Compounds From <i>Acalypha Indica</i> L. on MCF‐7 Breast Cancer Cells as Potential Anticancer Agents

Acalypha indica is a weed used in traditional cancer treatments. In this study, we present the first attempt to use a metabolomic approach to identify cytotoxic compounds from A. indica extract with potential anticancer properties. The leaves of A. indica were extracted using ethanol, ethyl acetate, chloroform, and n-hexane. The cytotoxic activity against MCF-7 cells was evaluated, revealing 50% inhibition concentration (IC₅₀) values ranging from 51.88 ± 7.80 to 226.86 ± 13.27 µg/mL. Metabolite profiling using liquid chromatography-high-resolution mass spectroscopy identified 27 metabolites and principal component analysis successfully differentiated the extracts, indicating variability in the compounds extracted using each solvent. To identify the cytotoxic compounds, orthogonal partial least-squares discriminant analysis was used to correlate metabolite profiles with IC₅₀ values from the cytotoxic assay. From the results obtained, we successfully predicted six compounds contributing to the anticancer activity of A. indica, namely hernanol, 13(S)-HpOTrE, (±)9-HpODE, (+)-catechin, traumatic acid, and one compound as assumed to be theobromine. In silico analysis predicted that (+)-catechin and hernanol bind to the alpha-estrogen receptor with an affinity similar to that of doxorubicin.