Preliminary Identification and Quantification of Four Secondary Metabolites, Total Tannin and Total Flavonoid Contents in Guava Fruit Ethanol Extract

Abstract

Introduction:The study on guava fruit ethanol extract from DukuhwaluhVillage, Purwokerto, Central Java, Indonesia showed increased megakaryocytes and platelet numbers in thrombocytopenic mice model. The study of acute oral toxicity of the extract did not show toxic effects in the kidney and liver at doses of 2000 and 5000 mg/kg b.w. The aim of the study was to determine the profile and quantity of four metabolite compounds and total tannin and flavonoid in the extract. Materials and Method: gallic acid, ellagic acid, rutin and kaemferol in the extract were identified and quantified by using high-performance liquid chromatography (HPLC) method with column LiChroCART 250-4,6 RP 18E, isocratic mobile phases with the composition of mixture:0.1% acetic acid, acetonitrile and methanol (40:50:10) respectively, and at temperature 300C. The total tannin and flavonoid were determined using the by spectrophotometry method, Fe(III) chloride and 1.10-phenanthroline at wavelength 510 nm for tannin and aluminium chloride and rutin at wavelength 422 nm for flavonoid, respectively. Results: Percentage of gallic acid, ellagic acid, rutin and kaempferol were 0.77%, 1.37 %, 0.41 % and 0.35 %, respectively. Total tannin and flavonoid contents were 1.20% (TAE) and, 1.18% (RE) respectively. Conclusion: The guava fruit ethanol extract contained gallic acid, ellagic acid, rutin, kaempferol, tannin and flavonoid.