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ZoP Characterization of the Bioactives from Mira-cle Fruit (Synsepalum dulcificum) against Klebsiella pneumoniae

autorzy: MARIA LEONORA L. DELOS SANTOS & CLINT ALLEN MARGALLO

fectious diseases are now at risk with the emerging mul-ti-drug resistant (MDR) pathogens such as Klebsiella pnuemoniae. The current investigation aims to charac-terize the bioactive compounds from Synsepalum dul-cificum leaves against Klebsiella pneumoniae. Liq-uid/liquid separation, open-column chromatography and thin layer chromatography was the method used in puri-fication. Bioautography was the method for screening the antibacterial activity of compounds present on TLC plate was conducted for bioassay against the test organ-ism. Characterization of the potent fraction was done us-ing infrared spectroscopy and nuclear magnetic reso-nance spectroscopy. Based on results F3 (135 ml tolu-ene: 15 ml acetone) and F4 (120 ml toluene: 30 ml ace-tone) fractions showed antibacterial activity in the bioau-togram after it was stained with MTT. Both fractions F3 & F4 have five positives (+++++ zone of inhibition is highly visible) meaning the compounds present in these fractions are effective against the test organism. Based on spectroscopic results, F3 contains both aromatic and aliphatic functions, with main absorptions of greater and below 3000 cm-1 . During NMR analysis,the sample was found to be impure thus the structure of the com-pounds cannot be elucidated yet. However, distinct sig-nals are still evident due to the 1H NMR signals at 7.40-7.43 ppm, 7.53-7.55 ppm, and 8.03-8.05 ppm. The sig-nals in this region are for the aromatic benzene.The sig-nal at 0.74-2.37 ppm indicate aliphatic methyls (CH3), methylenes (CH2), and methyne (CH) protons.The 13C spectrum also corroborated the signals of the 1H NMR. The peaks at 128.4 and 129.7 ppm indicate aromatic carbons. The peaks of 64.0 ppm and 69.2 ppm indicate methylene or methyne carbons which are directly at-tached to an oxygen or nitrogen. The result on bioassay proves that Synsepalum dulcificum leaves are potential source of antibiotics against K. pnuemoniae. Further pu-rification and spectroscopic analysis must be done to confirm the result of the study.