Isolation of scopoletin from ethyl acetate extract of Subang-Subang (Spilanthes paniculata Wall. Ex. DC.) was successfully established respectively by column and preparative thin layer chomatography. Ethyl acetate extract was separated by column chromatography using silica gel as stationary phase. Several organic solvent such as n- Hexane, ethyl acetate and methanol were used as mobile phase and the isolated compound was purified by preparative thin layer chromatography using n-Hexane : ethyl acetate (1:1) as the eluent. Isolated compound obtained was 11 mg of white solid form. Purification test of isolated compound which was effectued by TLC (Thin Layer Chromatography) using 10% NaOH gave blue colour fluorecence and the UV-Vis spectrum shown a weak absorption peak at 260 nm and 357 nm. Isolated compound which was characterized by FT-IR spectroscopic method indicated respectively the -OH , C-H, C=O, and C=C (aromatic) stretching at 3435 cm-1, 2917 cm-1, 1654 cm-1, 1535 cm-1, and only at 1462 cm-1 which was shown the C-H bending. Absorption bands at wave numbers 1241 cm-1 and 1165 cm-1 indicated the presence of ester groups. Based on the results of characterization, the isolated compound was classified into scopoletin compound. Bioactivity test as antibacterial of ethyl acetate extract and scopoletin compound by disk diffusion method had shown that the ethyl acetate extract and scopoletin compound are inactive against Neisseria sp bacteria.