Bio-Guided Study on Melastoma malabathricum Linn Leaves and Elucidation of its Biological Activities

The present study aimed to isolate and identify constituents with antibacterial activity from the Methanol extract of Melastoma malabathricum leaves (MMML) through bioassay guided fractionation. Furthermore, the study scoped to evaluate the cytotoxic effects of M. malabathricum leaves fractions for possible protective effect of antoxidative constituents present in the fractions. The MMML extract was fractionated by Vacuum Liquid Chromatography (VLC) to afford M. malabathricum leaves fractions (ML1-ML6) of increasing polarities. Antibacterial activity of plant extract, six fractions and two bioactive constituents against Staphylococcus aureus reference strain, methicillin-resistant Staphylococcus aureus reference strain (MRSA), eleven clinical MRSA isolates, three clinical Pseudomonas aeruginosa isolates and P. aeruginosa reference strain was evaluated using Minimum Inhibitory Concentration (MIC). The bioactive constituents responsible for antibacterial activity of MMML, fractions were detected by direct TLC-bioautography. The data obtained from MIC assay showed that ML5 effectively inhibited growth of all test bacterial pathogens. Direct TLC-bioautography revealed that ML5 had the highest number of antibacterial compounds. Following bioassay-guided fractionation, Kaempferol-3-O-(2", 6"-di-O-p-trans-coumaroyl)-β-glucopyranoside and Kaempferol were isolated from ML5. The structures of bioactive compounds were elucidated using Nuclear Magnetic Resonance (NMR) spectroscopy from 1HNMR and 13CNMR. In addition, the cytotoxic effect of M. malabathricum leaves fractions (ML1-ML6) revealed that ML5 had the highest Cytotoxicity Concentration (CC50) at 0.75 mg mL-1, an observation traceable to the presence of flavonoids constituents with antioxidant properties. Therefore, the results clearly indicate that MMML, fractions and bioactive constituents have profound antibacterial activity. Furthermore, antioxidative compounds from ML5 fraction exert their effects by enhancing the level of CC50.

• Publication year

  2013

• Venue

  American Journal of Applied Sciences

• Publication date

  2013-07-24

• Fields of study

  Biology, Medicine, Chemistry, Environmental Science

• Identifiers
  DOI 10.3844/AJASSP.2013.767.778
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• ML5 fraction showed the highest CC50 value, 0.75 mg mL-1, among the leaf fractions.

  Confidence 0.93

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• bioassay-guided fractionation of MMML yielded Kaempferol-3-O-(2", 6"-di-O-p-trans-coumaroyl)-β-glucopyranoside and Kaempferol from ML5 fraction, and NMR spectroscopy was used to elucidate their structures.

  Confidence 0.97

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• direct TLC-bioautography showed ML5 fraction contained the highest number of antibacterial compounds.

  Confidence 0.95

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• ML5 fraction inhibited growth of all tested bacterial pathogens in the MIC assay.

  Confidence 0.98

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review

• bioassay-guided fractionation

  method, process

  An iterative separation approach in which fractions are purified based on biological activity measurements.

  Aliases: bioassay guided fractionation

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• cytotoxicity concentration (cc50)

  measurement, assay outcome

  The concentration of a sample that produces 50% cytotoxicity in the reported cytotoxicity assay.

  Aliases: CC50

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• direct tlc-bioautography

  assay, method

  A thin-layer chromatography bioassay used to localize antibacterial spots directly on chromatographic plates.

  Aliases: TLC-bioautography

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• kaempferol

  compound, flavonoid

  A flavonoid compound isolated from ML5 fraction during the guided purification workflow.

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• kaempferol-3-o-(2", 6"-di-o-p-trans-coumaroyl)-β-glucopyranoside

  compound, flavonoid

  A flavonoid glycoside isolated from ML5 fraction during the guided purification workflow.

  Aliases: Kaempferol-3-O-(2'', 6''-di-O-p-trans-coumaroyl)-β-glucopyranoside

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• melastoma malabathricum leaves extract (mmml)

  extract, material

  The methanol extract prepared from Melastoma malabathricum leaves and used as the starting material for fractionation and bioactivity testing.

  Aliases: MMML

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• minimum inhibitory concentration (mic)

  assay, measurement

  The antibacterial assay used to determine the lowest concentration of an extract or fraction that prevents visible bacterial growth.

  Aliases: MIC

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• ml5 fraction

  fraction, material

  The VLC-derived leaf fraction with intermediate polarity among ML1-ML6 that was tested and further purified.

  Aliases: ML5

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• nuclear magnetic resonance (nmr) spectroscopy

  method, spectroscopy

  A spectroscopic technique used here with 1H and 13C NMR to determine the structures of isolated compounds.

  Aliases: NMR spectroscopy, 1H NMR, 13C NMR

  박진우 (dztg5apj7m) extractionAK (4715169a40) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review

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