Author: W. O Yuliastri, Sanatang, A. R. Syaban, W. O. G Purnamasari, M. Isrul, A. Yuhadi, R. Iswanto , A. Anzalna, R. A. A. Burhan, R. Yuniar and S. Ramadani

DOI: http://doi.org/10.31788/RJC.2026.1939667

This study investigated the immunomodulatory activity of Nypa fruticans Wurmb leaf fractions and characterisedtheir major bioactive constituents using LC–MS. The ethanolic leaf extract was successively fractionated withn- hexane, ethyl acetate, and ethanol, and each fraction was orally administered to male mice for 14 days. Immunomodulatory activity was evaluated through macrophage phagocytic activity, serum IL-12 and IFN-γ levels, and the expression of CD4 + , CD8 + , and CD14 + markers. All fractions significantly enhanced immune responsescompared with the negative control. The ethanol and ethyl acetate fractions showed the most consistent effects oncytokine production and lymphocyte modulation, whereas the n-hexane fraction produced the highest macrophagephagocytic activity. LC–MS profiling revealed that the active fractions were enriched with phenolic and flavonoidrelated compounds, including emodin-8-O-glucoside, quercitrin, and 2,4-dihydroxybenzaldehyde, which maycontribute to the observed immunostimulatory effects. These findings provide fraction-level scientific evidencesupporting N. fruticans as a promising source of plant-derived immunomodulatory agents and help bridge traditional medicinal use with modern pharmacological and phytochemical validation. The study also offers a foundationfor future investigations on compound isolation, mechanism of action, standardisation, and safety evaluation. Practically, phenolic-rich fractions of N. fruticans may be further developed as candidates for standardised herbal immunomodulators or supportive immune-enhancing therapies.

Keywords: Nypa fruticans Wurmb, Immunomodulatory Activity, Macrophage Phagocytosis, IL-12, IFN-γ, LC–MS, Phenolic Compounds, Flavonoids.