Bebuas (Premna serratifolia) traditionally is used by Malay people as a traditional medicine to restore health. This indicates it contains chemicals that potentially have an anti-inflammatory activity. The results of phytochemical screening showed that the ethanol extract contains phenolics, alkaloids, flavonoids, steroids and saponins. Meanwhile, hexane extract only contains steroids. Anti-inflammatory activity evaluation on crude extracts showed that the ethanol extract was more active than the hexane extract. Therefore, the isolation step was continued to ethanol extract with the targeted compound was an alkaloid. Extraction was conducted by using acid-base and partition techniques to give the alkaloids fraction (EaBW). EaBW is then further separated by liquid vacuum column technique using gradual polarity elution, with eluent of ethyl acetate (Ea) - methanol and obtained 5 fractions. The isolate was obtained from F.3. The isolate showed it had anti-inflammatory activity with ED50 = 5.45 mg/KgBW. The UV-Vis and FT-IR spectra patterns of the isolate have similarities with ciprofloxacin as well as its physical characteristics which are hygroscopic yellowish crystal, and fluorescence property. Therefore, the isolated compound is thought to have a similar structure to ciprofloxacin.