Osong Public Health Res Perspect.  2014 Oct;5(5):298-312. 10.1016/j.phrp.2014.08.001.

In Vitro Antibacterial Activity, Gas Chromatography–Mass Spectrometry Analysis of Woodfordia fruticosa Kurz. Leaf Extract and Host Toxicity Testing With In Vitro Cultured Lymphocytes From Human Umbilical Cord Blood

• Dubey D
• Patnaik R
• Ghosh G
• Padhy RN

• KMID: 2518155
• DOI: http://doi.org/10.1016/j.phrp.2014.08.001

Abstract

Objectives
To locate a plant with suitable phytochemicals for use as antimicrobial agents to control multidrug-resistant (MDR) bacteria as a complementary medicine, without host toxicity as monitored through cultured lymphocytes from human umbilical cord blood.
Methods
The methanol crude leaf extract of the plant Woodfordia fruticosa was subjected to antimicrobial assay in vitro with nine pathogenic MDR bacteria from clinical samples. This was followed by bioassay-guided fractionation with seven non-polar to polar solvents, gas chromatography–mass spectrometry analysis of the n-butanol fraction, and monitoring of the host toxicity of the leaf extract with in vitro grown lymphocytes from human umbilical cord blood.
Results
The leaf extract of W. fruticosa had a controlling capacity for MDR bacteria. The minimum inhibitory concentration and minimum bactericidal concentration of the n-butanol fraction were < 1.89 mg/mL extract and 9.63 mg/mL extract, respectively. The gas chromatography–mass spectrometry spectrum of the n-butanol fraction confirmed the presence of 13 peaks of different compounds with retention times of 9.11 minutes, 9.72 minutes, 10.13 minutes, 10.78 minutes, 12.37 minutes, 12.93 minutes, 18.16 minutes, 21.74 minutes, 21.84 minutes, 5.96 minutes, 12.93 minutes, 24.70 minutes, and 25.76 minutes. The six leading compounds were: diethyl phthalate: IUPAC name: diethyl benzene-1,2-dicarboxylate; 5-methyl-2-(1-methylethyl) phenol: IUPAC name: 5-methyl-2-propan-2-ylphenol; (E )-3,7-dimethylocta-2,6-diene-1-thiol: IUPAC name: (2Z)-3,7-dimethylocta-2,6-diene-1-thiol; 2,6,10-dodecatrien-1-ol, 3,7,11-trimethyl-, (E,E ): IUPAC name: 2,6,10-dodecatrien-1-ol; 3,7,11-trimethyl-, (E,E); 2-methoxy-4-(2-propenyl) phenol: IUPAC name: 2-methoxy-4-[(1E)-prop-1-en-1-yl]phenol; hexadecanoic acid: IUPAC name: hexadecanoic acid.
Conclusion
The presence of antimicrobial compounds that are therapeutically potent against MDR bacteria was confirmed in W. fruticosa. The crude leaf extract showed no host toxicity with human lymphocytes; the n-butanol fraction of the extract was the most suitable bioactive fraction. The terpenes isolated were: 5-methyl-2-(1-methylethyl) phenol, 2-methoxy-4-(2-propenyl) phenol, 2,6-octadien-1-ol, 3,7-dimethyl-(E)-2,6-octadienal, 3,7-dimethylcyclohexanol, and cyclohexanol, 2-methylene-5-(1-methylethenyl) which were reported to have specifically antimicrobial activity.

Keyword

gas chromatography–mass spectrometry analysis; human lymphocytes; multidrug-resistant bacteria