Bioassay-guided In vitro Study of the Antimicrobial and Cytotoxic Properties of the Leaves from Excoecaria Lucida Sw

摘要

Background: Excoecaria lucida Sw. ( Euphorbiaceae ) is a plant conventionally used throughout the Caribbean in the treatment of infectious diseases. Objective: To evaluate, using bioassay-guided fractionation, the in vitro cytotoxicity and antimicrobial activity of E. lucida leaves. Materials and Methods: A 95% ethanol crude extract was dried and fractionated by solid-liquid separation in four phases (hexane, dichloromethane, ethyl acetate, and butanol). Antimicrobial activity (3 bacteria, 6 yeasts, and 2 fungi) was evaluated by the dilution method with resazurin (2048, 512, 128, 32, and 8 μg/mL). The cytotoxicity assays were evaluated in two cell lines: MRC-5 and RAW 264.7; calculating the selectivity index. Assays were performed for the total extract, the isolated compound with the highest yield, and the ethyl acetate and butanol phases. Isolated compounds were characterized by nuclear magnetic resonance and mass spectrometry techniques. Results: Fractionation process led to the isolation of ellagic acid (784.29 mg), 3,3',4'-tri-O-methyl ellagic 4-O-β-D-glucopyranoside acid (6.1 mg), and corilagin (6.91 mg). The most active were ethyl acetate phase and ellagic acid with IC₅₀= 128 μg/mL against seven and five different species of microorganisms, respectively. The total extract (IC₅₀=512 μg/mL) and the ethyl acetate phase (IC₅₀=128 μg/mL) were cytotoxic in both cell lines, while butanol phase and ellagic acid both with IC₅₀₆: Deuterated dimethyl sulfoxide, LC.MS: Liquid Chromatography-Mass Spectrometry, [α]_D: Optical rotation, EL1: ellagic acid, EL2: 3,3’,4’-tri-O-methyl ellagic 4-O-β-D-glucopyranoside acid, EL3: corilagin, Active (+), inactive (-). Key words: Corilagin, ellagic acid, ethnopharmacological use, selectivity index, tanninsSUMMARY