Phytochemical Characterization and In Vitro Anti-Inflammatory, Antioxidant and Antimicrobial Activity of Combretum Collinum Fresen Leaves Extracts from Benin
Abstract
:1. Introduction
2. Results and Discussion
2.1. Structure Elucidation of Compounds in the 50% Ethanol Extract from C. collinum Leaves and Phytochemical Analysis of the n-Hexane Extract
2.2. In-Vitro Antioxidant Activity of C. collinum EtOH 50% Leaves Extract
2.3. In-Vitro Anti-Inflammatory Activity of C. collinum Aqueous Leaves Extract, Myricetin-3-O-rhamnoside and Myricetin
2.4. In-Vitro Antibacterial Activity of C. collinum EtOH 50% Leaves Extract
3. Materials and Methods
3.1. Plant Material
3.2. Phytochemical Analysis
3.2.1. GC-MS analysis of Hexane Extract
3.2.2. Chromatographic Separation and Isolation of Flavonoids
3.2.3. HPLC Analysis
3.2.4. Quantitative HPLC Analysis
3.2.5. LC-MS Analysis
3.2.6. NMR Experiments
3.2.7. DPPH Assay
3.2.8. ABTS Assay
3.3. Functional Investigations on HaCaT Cells
3.3.1. Material
3.3.2. Cell Culture
3.3.3. In-Vitro Anti-Inflammatory Assay
3.3.4. Cell Viability Assay
3.4. Antimicrobial Assays
3.4.1. Microbial Strains
3.4.2. Agar Well Diffusion Test
3.4.3. Microdilution Test
3.4.4. Bacterial Growth Curve
3.5. Data Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of Combretum collinum extracts and myricetin-3-O-rhamnoside are available from the authors. |
Peak | RT 1 [min] | λmax [nm] | m/z | Ion | Compound |
---|---|---|---|---|---|
1 | 20.177 | 264 350 | 319.096 481.189 479.193 | [A + H]+ [M + H]+ [M − H]− | Myricetin-3-O-glucoside |
2 | 23.219 | 262 350 | 319.094 465.179 463.188 | [A + H]+ [M + H]+ [M − H]− | Myricetin-3-O-rhamnoside |
Signal | Myricetin-3-O-rhamnoside (F31–F33) | Myricetin-3-O-rhamnoside [10] |
---|---|---|
H-3′′-6′′ | 0.95, d, J 6 Hz | 0.94, d, J 5.6 Hz |
sugar H | 3.3–3.8 | 3.3–3.8 |
H-2′′ | 4.25, s | 4.23, s |
H-1′′ | 5.34, brs | 5.31, brs |
H-6 | 6.22, s | 6.19, s |
H-8 | 6.39, s | 6.35, s |
H-2′ and H-6′ | 6.97, s | 6.95, s |
No. | RT 1 [min] | Compound | NIST Match Quality |
---|---|---|---|
1 | 12.62 | Malic acid | 97 |
2 | 12.73 | Tetradecanoic acid (syn. myristic acid) | 98 |
3 | 14.39 | Hexadecanoic acid (syn. palmitic acid) | 98 |
4 | 15.87 | Oleic acid | 98 |
5 | 15.92 | Octadecanoic acid (syn. stearic acid) | 99 |
6 | 17.64 | Eicosanoic acid (syn. arachidic acid) | 99 |
7 | 19.78 | Docosanoic acid (syn. behenic acid) | 99 |
8 | 22.40 | Tetracosanoic acid (syn. lignoceric acid) | 99 |
9 | 17.94 | Pentacosane | 96 |
10 | 18.99 | Hexacosane | 99 |
11 | 20.17 | Heptacosane | 99 |
12 | 21.48 | Octacosane | 99 |
13 | 22.92 | Nonacosane | 96 |
14 | 24.42 | Triacontane | 99 |
15 | 26.06 | Nonadecane | 96 |
16 | 23.22 | Squalene | 97 |
17 | 30.78 | Campestrol | 99 |
18 | 31.18 | Stigmasterol | 99 |
Species | Inhibition Zones (mm) | Inhibition Zones (Positive Controls) | MIC |
---|---|---|---|
S. epidermidis | 21.75 ± 0.43 | 28.67 ± 0.47 (G) | 275.0 µg/mL |
MRSA | 16.25 ± 0.43 | 18.67 ± 0.47 (V) | 385.5 µg/mL |
S. aureus | 15.75 ± 0.43 | 24.33 ± 0.47 (G) | n.d. |
K. pneumoniae | 14.00 ± 0.00 | 25.33 ± 0.94 (G) | n.d. |
P. aeruginosa | 11.00 ± 0.00 | 12.00 ± 0.00 (G) | n.d. |
E. coli | 11.25 ± 0.43 | 26.67 ± 0.47 (G) | n.d. |
Enterococcus | 0 | 16.67 ± 0.47 (G) | n.d. |
C. albicans | 0 | 26.67 ± 0.47 (N) | n.d. |
DMSO 10% | 0 | - | n.d. |
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Marquardt, P.; Seide, R.; Vissiennon, C.; Schubert, A.; Birkemeyer, C.; Ahyi, V.; Fester, K. Phytochemical Characterization and In Vitro Anti-Inflammatory, Antioxidant and Antimicrobial Activity of Combretum Collinum Fresen Leaves Extracts from Benin. Molecules 2020, 25, 288. https://doi.org/10.3390/molecules25020288
Marquardt P, Seide R, Vissiennon C, Schubert A, Birkemeyer C, Ahyi V, Fester K. Phytochemical Characterization and In Vitro Anti-Inflammatory, Antioxidant and Antimicrobial Activity of Combretum Collinum Fresen Leaves Extracts from Benin. Molecules. 2020; 25(2):288. https://doi.org/10.3390/molecules25020288
Chicago/Turabian StyleMarquardt, Peter, Rick Seide, Cica Vissiennon, Andreas Schubert, Claudia Birkemeyer, Virgile Ahyi, and Karin Fester. 2020. "Phytochemical Characterization and In Vitro Anti-Inflammatory, Antioxidant and Antimicrobial Activity of Combretum Collinum Fresen Leaves Extracts from Benin" Molecules 25, no. 2: 288. https://doi.org/10.3390/molecules25020288