Bioassay-Guided Separation Approach for Characterization of New Antibacterial Fractions from the Stem Roots Extracts of Archidendron jiringa

Abstract

Infectious diseases caused by bacteria has become a global health issue, especially antibacterial drug resistance. The most serious concern with antibacterial resistance is that some bacteria became resistant to almost all antibacterial drugs, which makes them less effective. Archidendron jiringa is one of the most potent medicinal plants to be developed as a new source of antibacterial components. In current study, based on the antibacterial assay-guided approach, the separation of bioactive fractions of A. jiringa stem roots was carried out through several stages including isolation, fractionation, and characterization. The stages of isolation of secondary metabolites were conducted by gradually extraction followed by fractionation using chromatographic methods. The antibacterial potential of extracts was evaluated by the disc diffusion and microdilution methods employing the resazurin assay against one Gram-negative resistant bacteria, Escherichia coli, and one Gram-positive bacteria, Bacillus subtilis. Among three extracts obtained, ethyl acetate and methanol extracts demonstrated the most significant antibacterial effects, while no antibacterial activity was showed on the hexane extract. The fractionation of ethyl acetate extract led to the isolation of the most bioactive fractions (E2815 and E2816) with the MIC’s values ranging of 12.5–25 µg/mL for both resistant bacteria. Due to less quantity, only the fraction E2816 was subjected to analysis by 1H-NMR spectroscopy. The results exhibited that the bioactive fraction was obtained as a mixture of at least three major constituents. However, the purification of the bioactive fraction is required, to further clarify the antibacterial compound that can be utilized as a new promising antibacterial agent. The bioassay-guided separation approach and the dye resazurin as an indicator of the growth of bacteria are applied for the first time for the phytopharmacological investigation from this plant. The present study represented the most effective method for subsequent finding and isolation of potential novel antibacterial constituents from A. jiringa stem roots, in particular against the multi-drug resistant strains.

Keywords

• A. jiringa
• antibacterial assay
• bioassay-guided separation
• medicinal plant

Supporting Institution

Universitas Lampung

Thanks

The authors would like to thank to the Institute of Research and Community Services, Universitas Lampung that provided fund for this project to be undertaken through Penelitian Kerjasama Internasional/International Research Collaboration 2019. The authors would like to thank Enago (www.enago.com) for the English language proofread and review.

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