Abstract
The Philippines is a megadiverse biodiversity hotspot featuring an extraordinary richness of endemic flora, much of which remains unexplored within modern bioprospecting frameworks. Concurrently, traditional indigenous healthcare systems maintain extensive empirical knowledge regarding the anti-inflammatory utility of local botanicals. This study presents an integrative workflow combining quantitative ethnomedical documentation with untargeted metabolomics to accelerate natural product drug discovery. Ethnobotanical surveys conducted among traditional healers in Luzon and Visayas prioritized three endemic species with documented anti-inflammatory applications: Cinnamomum mercadoi, Syzygium nitidum, and Goniothalamus amuyon. Methanolic leaf extracts were subjected to liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) metabolomic profiling paired with multivariate statistical modeling (PCA and OPLS-DA) and in vitro bioactivity testing on lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. Results demonstrated significant, concentration-dependent suppression of nitric oxide (NO) production and downregulated expression of pro-inflammatory cytokines (TNF-α and IL-6) by C. mercadoi and S. nitidum extracts (IC50 < 14.8 µg/mL) without inducing cytotoxicity. Feature annotation matched specialized metabolites, including specific proanthocyanidins, novel kaempferol glycosides, and secoiridoids, directly with observed anti-inflammatory activities. This research validates traditional medicinal applications through rigorous chemometric and biological frameworks, establishing a scalable, ethical model for bioprospecting that bridges indigenous knowledge systems, analytical chemistry, and pharmacological science.