Identification and Antibacterial Evaluation of Bioactive Compounds from Garcinia kola (Heckel) Seeds
Abstract
:1. Introduction
2. Results and Discussion
2.1. Susceptibility Testing
Zones of inhibition (mm) at different concentrations (mg/mL) | |||||
---|---|---|---|---|---|
Organism | Extract | 50 | 100 | 200 | Ciprofloxacin |
Ethyl Acetate | 21 ± 1.3 | 23 ± 0.7 | 20 ± 1.2 | 25 | |
Acetone | 19 ± 0.8 | 22 ± 2.3 | 23 ± 1.6 | 24 | |
S. pyogenes | Ethanol | 19 ± 0.6 | 19 ± 1.3 | 22 ± 0.9 | 24 |
Methanol | 20 ± 1.0 | 24 ± 1.1 | 21 ± 2.3 | 23 | |
Water | 0 | 0 | 0 | 25 | |
Ethyl Acetate | 17 ± 0.6 | 17 ± 1.6 | 14 ± 1.8 | 26 | |
Acetone | 19 ± 1.6 | 19 ± 0.6 | 21 ± 1.1 | 24 | |
S. aureus | Ethanol | 18 ± 1.5 | 23 ± 0.8 | 20 ± 0.3 | 27 |
Methanol | 21 ± 1.1 | 22 ± 2.3 | 19 ± 1.3 | 23 | |
Water | 0 | 0 | 0 | 24 | |
Ethyl Acetate | 0 | 0 | 0 | 30 | |
Acetone | 0 | 15 ± 1.3 | 12 ± 1.8 | 30 | |
P. shigelloides | Ethanol | 10 ± 2.3 | 19 ± 1.9 | 11 ± 2.3 | 31 |
Methanol | 18 ± 1.5 | 21 ± 1.3 | 19 ± 1.6 | 30 | |
Water | 0 | 0 | 0 | 31 | |
Ethyl Acetate | 0 | 0 | 0 | 30 | |
Acetone | 16 ± 1.7 | 14 ± 2.3 | 0 | 30 | |
S. typhimurium | Ethanol | 0 | 18 ± 1.3 | 14 ± 1.8 | 29 |
Methanol | 17 ± 0.6 | 18 ± 1.3 | 15 ± 1.2 | 30 | |
Water | 0 | 0 | 0 | 31 |
2.2. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) Determination
Organism | MIC values of Extract (mg/mL) | MIC values of Ciprofloxacin(mg/mL) | MBC values of Extracts(mg/mL) | MBC values of Ciprofloxacin (mg/mL) |
---|---|---|---|---|
S. pyogenes | 0.04 | 0.0012 | 0.081 | 0.0781 |
S. aureus | 0.04 | 0.0024 | 0.25 | 0.1563 |
P. shigelloides | 1.25 | 0.0049 | 2.5 | 0.3125 |
S. typhimurium | 0.63 | 0.0195 | 1.25 | 0.1563 |
2.3. Phytochemical Analysis
2.3.1. Thin Layer Chromatography (TLC)
Solvent systems | |||||||
---|---|---|---|---|---|---|---|
Bands | BEA | CEF | EMW | ||||
50 | 100 | 50 | 100 | 50 | 100 | ||
1 | 0.08 | 0.01 | 0.09 | 0.08 | 0.03 | 0.09 | |
2 | 0.1 | 0.13 | 0.14 | 0.11 | 0.08 | 0.16 | |
3 | 0.12 | 0.16 | 0.39 | 0.19 | 0.11 | 0.19 | |
4 | - | 0.22 | 0.5 | 0.39 | 0.16 | 0.31 | |
5 | - | 0.28 | 0.52 | 0.53 | 0.31 | 0.49 | |
6 | - | 0.32 | 0.53 | 0.55 | 0.49 | 0.56 | |
7 | - | - | 0.63 | 0.63 | 0.56 | 0.64 | |
8 | - | - | 0.72 | 0.64 | 0.72 | 0.67 | |
9 | - | - | 0.83 | 0.72 | 0.75 | 0.76 | |
10 | - | - | 0.87 | 0.83 | - | 0.88 | |
11 | - | - | - | 0.85 | - | - | |
12 | - | - | - | 0.88 | - | - |
2.3.2. Antimicrobial Activity Assay by Bioautography
Organisms | a | b | Solvent system | ||
---|---|---|---|---|---|
(50 mg/mL | 100 mg/mL) | (50 mg/mL | 100 mg/mL) | ||
S. pyogenes | 0.53 | 0.53 | +++ | +++ | CEF |
0.64 | 0.64 | +++ | +++ | CEF | |
0.30 | 0.31 | + | + | EMW | |
origin | Origin | ++++ | ++++ | BEA | |
S. aureus | - | 0.53 | - | + | CEF |
0.56 | 0.63 | +++ | +++ | EMW | |
- | - | - | - | BEA | |
P. shigelloides | 0.63 | 0.63 | ++ | ++ | CEF |
0.73 | 0.72 | ++ | +++ | CEF | |
- | 0.85 | - | ++ | EMW | |
0.75 | 0.76 | ++ | ++ | EMW | |
- | 0.88 | - | ++ | EMW | |
- | - | - | - | BEA | |
S. typhimurium | 0.53 | 0.55 | ++ | ++ | CEF |
0.63 | 0.64 | ++++ | +++ | CEF | |
0.52 | 0.52 | ++ | +++ | EMW | |
origin | Origin | ++ | ++ | BEA |
2.4. Column Chromatography Analysis and MIC90 Determination of Fractions
Fraction | Rf value of fractionated compounds | MIC90 against test organisms | |||
---|---|---|---|---|---|
S. pyogenes | S. aureus | P. shigelloides | S. typhimurium | ||
C 1 | 0.008, 0.159, 0.31 | ND | ND | ND | ND |
C2 | 0.09, 0.12, 0.36 | ND | ND | ND | ND |
C3 | 0.13, 0.17, 0.29 | 0.0195 | 0.00781 | ND | ND |
C4 | 0.156, 0.286, 0.294 | 0.00871 | 0.0195 | ND | ND |
CEF 1 | 0.2, 0.366, 0.42 | 0.049 | 0.0098 | 0.625 | 0.3125 |
CEF 2 | 0.34, 0.4, 0.52 | 0.625 | 0.024 | 0.625 | 1.25 |
CEF 3 | 0.153 | 0.1563 | 0.0781 | 0.0781 | 0.1563 |
CEF 4-6 | 0.13, 0.226, 0.3 | ND | 0.024 | 2.5 | 2.5 |
CEF 7 | 0.2, 0.33 | 0.625 | 0.0049 | ND | 0.1563 |
CEF 8-10 | 0.16, 0.306, 0.36 | 0.0195 | 0.024 | 2.5 | 2.5 |
CEF 11 | 0.13 | 0.049 | 0.0006 | 2.5 | 0.012 |
CEF 12 | 0.15, 0.18 | 0.3125 | 0.635 | 0.0781 | 1.25 |
CEF 13 | 0.13, 0.18, 0.28 | 0.0049 | 0.024 | 1.25 | ND |
CEF 14-16 | 0.15, 0.2 | 0.3125 | 1.25 | 1.625 | ND |
CEF 17 | 0.11, 0.38 | 0.0024 | 0.049 | 0.325 | ND |
CEF 18 | 0.12, 0.27 | 0.0012 | 0.1563 | 0.0195 | ND |
Ciprofloxacin | 0.0012 | 0.0049 | 0.012 | 0.0781 |
2.5. Gas-Chromatography/Mass-Spectrometry (GC-MS)
Fraction | Peak numbers | RT (mins) | Compounds identified | % |
---|---|---|---|---|
1 | 29.89 | Hexadecanoic acid, methyl ester | 0.73 | |
2 | 30.42 | Hexadecanoic acid | 4.41 | |
3 | 31.93 | Hexadecanoic acid, Palmitic acid | 25.07 | |
4 | 31.22 | Hexadecanoic acid, ethyl ester | 0.83 | |
CEF 3 / F3 | 5 | 33.19 | 9-Octadecenoic acid, methyl ester | 0.91 |
6 | 33.71 | Heptadecene-(8)-carbonic acid | 6.63 | |
7 | 34.10 | Linoloic acid | 26.60 | |
8 | 34.24 | 9-Octadecenoic acid | 24.81 | |
9 | 34.42 | 14-Pentadecanoic acid | 2.62 | |
10 | 41.20 | Octadenoic acid, Stearic acid | 7.49 | |
Total | 100 | |||
CEF 11 / F11 | 1 | 41.20 | 1.2-Benzenedicarboxylic acid | 100 |
1 | 6.63 | Formamide, N,N-Diethyl | 2.31 | |
2 | 8.45 | 1-Butanol | 15.72 | |
3 | 9.95 | 3-Isothiazolecarboxamide | 3.03 | |
4 | 12.07 | 2.3-Dihydro-3,5-dihydeoxy-6-methyl ester | 24.16 | |
5 | 15.04 | 2.5-Di(hydroxymethyl)-furan | 7.06 | |
6 | 30.78 | Palmitic acid | 12.91 | |
CEF 12 / F12 | 7 | 33.94 | 9-Octadecanoic acid | 4.08 |
8 | 34.02 | Cyclohexadecane | 1.84 | |
9 | 34.36 | Stearic acid | 3.11 | |
10 | 34.68 | n-Tetradecanoic acid amide | 2.55 | |
11 | 37.75 | 9-Octadecenamide | 13.82 | |
12 | 38.11 | Hexadecanamide | 1.59 | |
13 | 40.34 | 3, 4,8-Trimethyl-2-nonenal | 2.07 | |
Total | 94.24 |
3. Experimental
3.1. Bacterial Strains
3.2. Resuscitation of Bacterial Strains
3.3. Preparation of Plant Extract
3.4. Antimicrobial Susceptibility Testing
3.5. Determination of Minimum Inhibitory Concentration (MIC90)
3.6. Determination of Minimum Bactericidal Concentration (MBC)
3.7. Phytochemical and Antimicrobial Analysis
3.7.1. Thin Layer Chromatography (TLC)
3.7.2. Antimicrobial Activity Assay by Bioautography
3.8. Column Chromatography
3.9. Determination of Minimum Inhibitory Concentration (MIC90) of Fractions
3.10. Gas-Chromatography/Mass-Spectrometry (GC-MS)
3.11. Statistical Analysis
4. Conclusions
Acknowledgements
References and Notes
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Seanego, C.T.; Ndip, R.N. Identification and Antibacterial Evaluation of Bioactive Compounds from Garcinia kola (Heckel) Seeds. Molecules 2012, 17, 6569-6584. https://doi.org/10.3390/molecules17066569
Seanego CT, Ndip RN. Identification and Antibacterial Evaluation of Bioactive Compounds from Garcinia kola (Heckel) Seeds. Molecules. 2012; 17(6):6569-6584. https://doi.org/10.3390/molecules17066569
Chicago/Turabian StyleSeanego, Christinah T., and Roland N. Ndip. 2012. "Identification and Antibacterial Evaluation of Bioactive Compounds from Garcinia kola (Heckel) Seeds" Molecules 17, no. 6: 6569-6584. https://doi.org/10.3390/molecules17066569