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Purpose: To investigate the anti-inflammatory activity of the ethanol extract of the immature fruit of a citrus, Shiranuhi, and to identify the active ingredient. Methods: The immature Shiranuhi peel was extracted with 80 % ethanol, and the extract was fractionated with solvents (n-hexane, ethyl acetate and n-butanol) to afford the corresponding fractions and water residue. Among them, the EtOAc-soluble portion was subjected to medium pressure liquid chromatography (MPLC) over a reversed-phase SiO 2 column to give compound 1 . The isolated compound was identified based on the proton and carbon nuclear magnetic resonance (NMR) spectra. The release of nitric oxide, prostaglandin (PG)E2, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 secreted by mouse macrophages was measured using RAW264.7 cell culture supernatant. Results: Shiranuhi (Korean name, Hallabong) is an important citrus species cultivated in Jeju Island, Korea. A polymethoxyflavonoid (PMF), tetramethyl-O-scutellarin ( 1 ), was isolated from the peels of immature Shiranuhi fruit. Upon the evaluation of anti-inflammatory effects, the flavonoid 1 decreased the nitric oxide production in macrophage cells with high efficiency, viz, 50 % inhibition concentration, IC 50 of 57.4 μM. Subsequent studies demonstrated that PMF 1 effectively inhibited the generation of PGE2, TNF-α, IL-1β, and IL-6 cytokine in a dose-dependent manner. Conclusion: Tetramethyl-O-scutellarin ( 1 ) has been successfully isolated from Shiranuhi species for the first time. Thus, Shiranuhi fruit peel extract containing PMF 1 can potentially be applied as an antiinflammatory ingredient in food or cosmetic industries. Keywords: Shiranuhi fruit, Nitric oxide, Tetramethyl-O-scutellarin, Anti-inflammatory

Tetramethyl-O-scutellarin isolated from peels of immature Shiranuhi fruit exhibits anti-inflammatory effects on LPSinduced RAW264.7 cells