Isolation and characterization of antibacterial compounds from five selected plants used against bacteria which infects wounds

Abstract

Five plant species: Ziziphus mucronata, Senna italica, Lantana camara, Ricinus communis and Lippia javanica, were selected for this study based on their use in traditional medicine. In preliminary screening, crude extracts were prepared using hexane, dichloromethane (dcm), acetone and methanol. Phytochemical profiles on Thin Layer Chromatography plates of the extracts were obtained by developing the plates in mobile phases of varying polarity. Tests for compounds such as tannins, flavonoids, alkaloids, phlobatannins, terpenes, steroids, cardiac glycosides and saponins were carried out. Antibacterial activity of the extracts was carried out using microdilution assay for Minimum Inhibitory Concentration and bioautography against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis. Antioxidant activity of the extracts was performed using the 2, 2, diphenyl-1-picrylhydrazyl (DPPH) assay. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay and Phagoburst test were used to investigate the toxic effects and anti-inflammatory activity of the extracts on mouse Raw 264.7 macrophage cells, respectively. The presence of phytochemicals was observed on the chromatograms after the plates were sprayed with vanillin sulphuric acid reagent. The dcm extracts of the plants showed antibacterial activity against the selected bacterial species on the bioautograms. Senna italica and Z. mucronata showed the most activity bands on the bioautograms. Lippia javanica had the lowest MIC average of 0.56 mg/ml. Antioxidant activity was observed in the extracts of L. javanica and R. communis. The extracts promoted proliferation of the mouse macrophage cells Raw 264.7 at concentrations ranging from 0.31 mg/ml and 0.08 mg/ml. Senna italica leaves were selected for isolation of antibacterial compounds. The isolated compound was analysed on 1H and 13C nuclear magnetic resonance (NMR) and Mass Spectrometry (MS) for structural analysis. The structure could not be elucidated due to impurities in the compound but the tentative structure is a branched chain alkane with at least one ether linkage per repeating unit. Therefore the study shows that there are plant components with biological activities against wound infecting bacteria and a single lead compound was identified.