The flavonoids derived from Eugenia polyantha leaves were characterized by rapid and low cost approach. The aim of this research is to characterize secondary metabolite profile of flavonoids in the n-butanol fraction of E. polyantha using thin layer chromatography and ultraviolet-visible spectrophotometry. The n-butanol fraction was separated by using silica gel 60 GF254 as the stationary phase; chloroform: ethanol: glacial acetic acid (9.4: 0.5: 0.1) as the mobile phase; and visualized by using UV light 366 nm. Five isolates were obtained from the separation, but there were only three isolates (Rf 0.26; 0.44; 0.77) respectively identified as flavonoid compounds. The characterization of the isolates by UV-Vis spectrophotometry showing that the ranges of λmax were 250-280 nm (band II) and 310-360 nm (band I), which indicate the existence of flavone compounds. Further characterization of the three isolate using the AlCl3 5%/HCl 6 M solution showed that the λmax shifted from band I to the higher wavelength (bathochromic). The λmax shift indicated the existences of ketone at C-4, hydroxyl group at C-5, and orthodihydroxyl at ring B. According to the maximum wavelength, the result of the characterization showed that the flavonoid compounds of the n-butanol were 5,3’,4’-trihydroxyflavone3-C-glycoside or 5,4’,5’-trihydroxyflavone-3-C-glycoside; 5,6,3’,4’-tetrahydroxyflavone or 5,6,4’,5’-tetrahydroxyflavone; and 5,3’,4’-trihydroxyflavone or 5,4’,5’-trihydroxyflavone.



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