Stachys Species: Comparative Evaluation of Phenolic Profile and Antimicrobial and Antioxidant Potential
Abstract
:1. Introduction
2. Results and Discussions
2.1. Chromatographic Analysis
2.2. Antioxidant Activity of Stachys sp. Extracts
2.3. Antimicrobial Activity of Stachys sp. Extracts
2.3.1. Antibacterial Activity Using the Agar-Well Diffusion Method
2.3.2. Antibacterial Activity Using the Broth Microdilution Method
2.3.3. Antibiofilm Activity
3. Materials and Methods
3.1. Plant Material
3.2. Chemicals
3.3. Preparation of Stachys sp. Extracts
3.4. LC-MS/MS Analysis of Polyphenols
3.5. LC-MS/MS Analysis of Rosmarinis Acid
3.6. Total Polyphenols Content
3.7. Flavonoid Content
3.8. Caffeic Acid Derivative Content
3.9. Antioxidant Activity
3.9.1. DPPH Radical Scavenging Assay
3.9.2. FRAP Assay
3.9.3. Nitrite-Induced Autooxidation of Hemoglobin
3.9.4. Inhibition of Lipid Peroxidation Catalyzed by Cytochrome c
3.9.5. Free Radical Generation Experiment
3.10. Determination of Antimicrobial Activity
3.10.1. Agar-Well Diffusion Method
3.10.2. Broth Microdilution Method
3.10.3. Anti-Biofilm Assay
3.11. Statistical Data Analysis
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Phenolic Compounds | m/z Value (Precursor) | m/z Value (Daughter Ions; Collision Energy) | tR ± SD (min) | S. officinalis | S. germanica | S. byzantina | S. sylvatica | S. palustris | S. recta |
---|---|---|---|---|---|---|---|---|---|
Gallic acid | 169 | 169 (0.5 V) | 1.50 ± 0.01 | - | - | - | - | 141.5 ± 8.4 | - |
Rosmarinic acid | 359 | 160.7; 178.6; 196.7 (0.9 V) | 2.20 ± 0.01 | 43.4 ± 0.5 | 7.8 ± 0.01 | 3.0 ± 0.02 | 1.9 ± 0.02 | 2.4 ± 0.01 | 2.6 ± 0.05 |
Protocatechuic acid | 153 | 153 (0.5 V) | 2.80 ± 0.01 | 94.6 ± 0.5 | 83.5 ± 0.4 | 28.6 ± 0.3 | 100.7 ± 9.2 | 94.2 ± 1.7 | 157.8 ± 2.1 |
Gentisic acid | 153 | 108.7 (0.9 V) | 3.52 ± 0.04 | <0.2 | <0.2 | <0.2 | <0.2 | <0.2 | <0.2 |
Caftaric acid | 311 | 148.6; 178.6 (0.9 V) | 3.54 ± 0.05 | 4.7 ± 0.2 | - | - | - | - | - |
Chlorogenic acid | 353 | 178.7; 190.7 (0.9 V) | 5.62 ± 0.05 | 1131.8 ± 15.8 | 4205.2 ± 44.7 | 2012.1 ± 37.8 | 5552.7 ± 47.2 | 3595.2 ± 24.7 | 6761.4 ± 58.5 |
Vanillic acid | 167 | 167 (0.5 V) | 6.70 ± 0.01 | 83.5 ± 0.7 | 21.1 ± 0.1 | 16.4 ± 0.3 | 16.6 ± 0.2 | 6.2 ± 0.08 | 20.3 ± 0.5 |
Syringic acid | 197 | 197 (0.5 V) | 8.40 ± 0.01 | 82.7 ± 0.2 | - | 7.0 ± 0.06 | 8.6 ± 0.1 | 4.4 ± 0.3 | - |
p-Coumaric acid | 163 | 118.7 (0.9 V) | 9.48 ± 0.08 | 27.8 ± 0.1 | 6.2 ± 0.08 | 4.4 ± 0.08 | 6.2 ± 0.3 | 14.0 ± 0.4 | 32.0 ± 0.9 |
Ferulic acid | 193 | 133.7; 148.7; 177.6 (0.9 V) | 12.80 ± 0.10 | 11.1 ± 0.4 | 5.0 ± 0.02 | 3.5 ± 0.04 | 3.0 ± 0.06 | 6.0 ± 0.2 | <0.2 |
Isoquercitrin | 463 | 254.9; 270.9; 300.7; 342.8 (1.2 V) | 19.60 ± 0.1 | 18.9 ± 0.1 | 233.1 ± 6.8 | - | 86.2 ± 3.7 | - | 72.8 ± 2.1 |
Rutin | 609 | 254.9; 270.9; 300.7; 342.8 (1.2 V) | 20.20 ± 0.15 | <0.2 | - | - | - | - | - |
Quercitrin | 447 | 178.8; 300.7 (1.2 V) | 23.64 ± 0.13 | - | - | - | - | <0.2 | - |
Luteolin | 285 | 150.6; 174.6; 198.6; 240.7 (1.5 V) | 29.10 ± 0.19 | 15.7 ± 0.2 | 26.8 ± 0.1 | 12.2 ± 0.1 | <0.2 | 3.9 ± 0.2 | <0.2 |
Apigenin | 269 | 148.6; 150.6; 224.7; 226.7 (1.5 V) | 33.10 ± 0.15 | 84.2 ± 1.7 | 21.4 ± 0.2 | 27.3 ± 0.6 | 13.6 ± 0.2 | <0.2 | <0.2 |
Stachys sp. Extracts | TPC (mg GAE/mL) | Flavonoids (mg RE/mL) | Caffeic Acid Derivatives (mg CAE/mL) |
---|---|---|---|
S. officinalis (SO) | 2.51 ± 0.09 a | 1.18 ± 0.01 d | 0.20 ± 0.01 g |
S. germanica (SG) | 5.98 ± 0.21 a | 3.30 ± 0.11 f | 0.88 ± 0.01 i |
S. byzantina (SB) | 6.40 ± 0.11 | 3.90 ± 0.21 | 0.89 ± 0.05 |
S. sylvatica (SS) | 4.00 ± 0.13 a | 1.22 ± 0.06 d,f | 0.20 ± 0.03 h |
S. palustris (SP) | 5.35 ± 0.29 b,c | 3.14 ± 0.17 f | 0.72 ± 0.06 g,i |
S. recta (SR) | 3.78 ± 0.17 a,c | 1.13 ± 0.09 e | 0.23 ± 0.08 h,i |
Stachys sp. Extracts | DPPH (IC50 µg/mL) | FRAP (μM TE/mL) | NHA mg CATE/g |
---|---|---|---|
S. officinalis | 139.16 ± 3.80 a | 280.17 ± 1.83 b | 23.44 ± 3.24 |
S. germanica | 54.18 ± 1.16 a | 688.21 ± 5.91 | 66.59 ± 5.37 |
S. byzantina | 53.61 ± 1.19 a | 354.88 ± 3.11 b | 27.06 ± 3.00 |
S. sylvatica | 76.04 ± 0.35 a | 218.01 ± 0.98 b | 20.01 ± 6.78 |
S. palustris | 54.82 ± 1.33 a | 502.43 ± 5.56 b | 59.38 ± 2.54 |
S. recta | 84.94 ± 1.45 a | 305.15 ± 2.84 b | 37.49 ± 1.96 |
Trolox | 11.19 ± 0.09 | - | - |
Inhibition Zone Diameter (IZD, mm) | |||||
---|---|---|---|---|---|
Stachys sp. Extracts | Staphylococcus aureus | Listeria monocytogenes | Escherichia coli | Salmonella enteritidis | Candida albicans |
S. officinalis | 16 ± 0.50 b | 14 ± 1.00 a | 10 ± 1.00 a | 10 ± 0.25 a | 12 ± 0.50 d |
S. germanica | 20 ± 0.25 a | 18 ± 0.50 a | 8 ± 0.25 a | 10 ± 0.50 a | 10 ± 0.50 d |
S. byzantina | 20 ± 0.25 a | 18 ± 0.00 a | 8 ± 0.50 a | 8 ± 0.50 a | 12 ± 1.00 d |
S. sylvatica | 16 ± 0.75 b | 16 ± 0.25 a | 8 ± 1.00 a | 8 ± 1.00 a | 10 ± 0.50 d |
S. palustris | 18 ± 0.50 c | 16 ± 1.00 a | 8 ± 0.00 a | 8 ± 0.25 a | 12 ± 0.50 d |
S. recta | 16 ± 0.00 b | 16 ± 1.00 a | 10 ± 0.25 a | 12 ± 0.50 b | 12 ± 0.00 d |
Gentamicin | 18 ± 0.25 | 22 ± 0.50 | 18 ± 0.25 | 19 ± 1.00 | - |
Fluconazole | - | - | - | - | 21 ± 0.00 |
Stachys sp. Extracts | Staphylococcus aureus | Listeria monocytogenes | Escherichia coli | Salmonella enteriditis | Candida albicans | |||||
---|---|---|---|---|---|---|---|---|---|---|
MIC | MBC | MIC | MBC | MIC | MBC | MIC | MBC | MIC | MFC | |
S. officinalis | 0.705 | 1.411 | 1.411 | 1.411 | 1.411 | 1.411 | 1.411 | 1.411 | 0.352 | 1.411 |
S. germanica | 0.423 | 0.423 | 0.847 | 1.695 | 3.39 | 3.39 | 3.39 | 3.39 | 1.695 | 3.39 |
S. byzantina | 0.468 | 0.468 | 1.875 | 3.75 | 3.75 | 3.75 | 3.75 | 3.75 | 0.937 | 3.75 |
S. sylvatica | 1.20 | 2.40 | 1.20 | 2.40 | 2.40 | 2.40 | 2.40 | 2.40 | 1.20 | 2.40 |
S. palustris | 0.817 | 1.635 | 1.635 | 3.27 | 3.27 | 3.27 | 3.27 | 3.27 | 0.817 | 3.27 |
S. recta | 1.12 | 2.24 | 1.12 | 2.24 | 2.24 | 2.24 | 2.24 | 2.24 | 0.56 | 2.24 |
Gentamicin MIC (mg/L) | 3 | 4 | 3 | 3 | - | |||||
Fluconazole MIC (mg/L) | - | - | - | - | 8 |
% Inhibition | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|
Stachys sp. Extracts | Staphylococcus aureus | Listeria monocytogenes | Escherichia coli | Salmonella enteriditis | Candida albicans | |||||
T0 | T24 | T0 | T24 | T0 | T24 | T0 | T24 | T0 | T24 | |
S. officinalis | + | - | + | - | + | - | - | + | ++ | + |
S. germanica | ++ | ++ | ++ | + | + | - | - | + | + | + |
S. byzantina | ++ | ++ | + | + | + | - | - | ++ | ++ | + |
S. sylvatica | ++ | + | + | + | ++ | + | - | - | ++ | - |
S. palustris | ++ | + | ++ | + | + | - | - | + | ++ | - |
S. recta | + | + | + | - | + | - | - | - | ++ | - |
Gentamicin | + | ++ | + | ++ | - | ++ | - | ++ | - | - |
Fluconazole | - | - | - | - | - | - | - | - | ++ | ++ |
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Benedec, D.; Oniga, I.; Hanganu, D.; Tiperciuc, B.; Nistor, A.; Vlase, A.-M.; Vlase, L.; Pușcaș, C.; Duma, M.; Login, C.C.; et al. Stachys Species: Comparative Evaluation of Phenolic Profile and Antimicrobial and Antioxidant Potential. Antibiotics 2023, 12, 1644. https://doi.org/10.3390/antibiotics12111644
Benedec D, Oniga I, Hanganu D, Tiperciuc B, Nistor A, Vlase A-M, Vlase L, Pușcaș C, Duma M, Login CC, et al. Stachys Species: Comparative Evaluation of Phenolic Profile and Antimicrobial and Antioxidant Potential. Antibiotics. 2023; 12(11):1644. https://doi.org/10.3390/antibiotics12111644
Chicago/Turabian StyleBenedec, Daniela, Ilioara Oniga, Daniela Hanganu, Brîndușa Tiperciuc, Adriana Nistor, Ana-Maria Vlase, Laurian Vlase, Cristina Pușcaș, Mihaela Duma, Cristian Cezar Login, and et al. 2023. "Stachys Species: Comparative Evaluation of Phenolic Profile and Antimicrobial and Antioxidant Potential" Antibiotics 12, no. 11: 1644. https://doi.org/10.3390/antibiotics12111644