Isolation and activity of two antibacterial biflavonoids from leaf extracts of Garcinia livingstonei (Clusiaceae).

The antibacterial activity of the acetone extract of Garcinia livingstonei leaves was studied using bioautography and by determining the minimum antibacterial concentration against four nosocomial pathogens. Bioautograms showed that two compounds were mainly responsible for the antibacterial activity. These were isolated by a combination of solvent-solvent fractionation and bioassay-guided chromatographic fractionation and were characterized by 1H NMR, 13C NMR and 2D NMR spectroscopy as amentoflavone and 4"-methoxy amentoflavone. The antibacterial activity of the isolated compounds was determined against Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeruginosa. Three of the tested organisms were sensitive to both compounds with MIC values ranging from 8-100 microg/mL. P. aeruginosa was resistant with MICs > 100 microg/mL. The safety of the two compounds was assessed with a tetrazolium based colorimetric assay (MTT assay) using Vero monkey kidney cells. The compounds had low toxicity against the cell line with cytotoxic concentrations to 50% of the cells (LD50) of 386 microg/mL and > 600 microg/mL for amentoflavone and 4"-methoxy amentoflavone, respectively. Berberine, the positive control had a CC50 of 170 microg/mL. 4"-Methoxy amentoflavone was more active and much less toxic than amentoflavone.