Antimicrobial and antioxidant properties of green synthesis CuO nanoparticles using Salvia officinalis hydroalcoholic extract
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Abstract
Background and Objectives: Following the emergence of microbial resistance to chemical antimicrobial agents, researchers have recently focused on the effects of nanoparticles and their antimicrobial activity. This research aimed to investigate the antimicrobial and antioxidant activity of copper oxide nanoparticles (CuO-NPs) produced using the hydroalcoholic extract of Salvia officinalis (S. officinalis).
Materials and Methods: In this study, after preparing the hydroalcoholic extract of S. officinalis and synthesizing CuO-NPs using this extract, the synthesized nanoparticles were evaluated. The antimicrobial properties of synthesized CuO-NPs were investigated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using the microbroth dilution method. The minimum biofilm inhibitory concentration (MBIC) and antioxidant activity of CuO-NPs and the hydroalcoholic extract of S. officinalis against Pseudomonas aeruginosa (P. aeruginosa) were investigated.
Results: CuO-NPs have a size in the range of 40 nm and a monoclinic crystal structure. Antimicrobial activity assessment by well diffusion showed that the synthesized CuO-NP nanoparticles at concentrations below 5 mg/mL had no antimicrobial effect on any of the bacteria studied. The lowest MIC of the synthesized CuO-NPs was observed against Staphylococcus saprophyticus (S. saprophyticus), and the highest against P. aeruginosa and Bacillus cereus (B. cereus). On the other hand, the highest MBC was related to B. cereus. The plant extract exhibited much weaker antimicrobial activity than CuO-NPs. The results obtained showed that the MBIC for CuO-NPs was 312 μg/mL. The half-maximal inhibitory concentration (IC50) obtained from CuO-NPs and the hydroalcoholic extract of S. officinalis was 2.79 and 1.97mg/ml, respectively.
Conclusion: Based on the results of this study, the green synthesis of CuO-NPs using hydroalcoholic extract of S. officinalis is a suitable candidate for the preparation of antimicrobial and antibiofilm compounds that also have antioxidant properties.
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2. Saxena A, Tripathi R, Zafar F, Singh P. Green synthesis of silver nanoparticles using aqueous solution of Ficus benghalensis leaf extract and characterization of their antibacterial activity. Mater Lett 2012; 67: 91-94.
3. Gour A, Jain NK. Advances in green synthesis of nanoparticles. Artif Cells Nanomed Biotechnol 2019; 47: 844-851.
4. Hussain I, Singh N, Singh A, Singh H, Singh S. Green synthesis of nanoparticles and its potential application. Biotechnol Lett 2016; 38: 545-560.
5. Villagrán Z, Anaya-Esparza LM, Velázquez-Carriles CA, Silva-Jara JM, Ruvalcaba-Gómez JM, Aurora-Vigo EF, et al. Plant-based extracts as reducing, capping, and stabilizing agents for the green synthesis of inorganic nanoparticles. Resources 2024; 13: 70.
6. Jadoun S, Arif R, Jangid NK, Meena R. Green synthesis of nanoparticles using plant extracts: A review. Environ Chem Lett 2021; 19: 355-374.
7. Nadaroglu H, GÜNGÖR AA, Selvi İ. Synthesis of nanoparticles by green synthesis method. Int J Innov Res Rev 2017; 1: 6-9.
8. Grdiša M, Jug-Dujaković M, Lončarić M, Carović-Stanko K, Ninčević T, Liber Z, et al. Dalmatian sage (Salvia officinalis L.): A review of biochemical contents, medical properties and genetic diversity. Agric Conspec Sci 2015; 80: 69-78.
9. Lu Y, Foo LY. Polyphenolics of Salvia—a review. Phytochemistry 2002 ; 59: 117-140.
10. Walch SG, Kuballa T, Stühlinger W, Lachenmeier D. Determination of the biologically active flavour substances thujone and camphor in foods and medicines containing sage (Salvia officinalis L.). Chem Cent J 2011; 5: 44.
11. Khakpour S, Khosravi M, Jafari Marandi S, Ahadi Ali M. The effect of hydroalcoholic extract of Salvia officinalis L. on the inflammatory reduction in male mice. AMU-TMUJ 2014; 24: 136-142.
12. Singh R, Shedbalkar UU, Wadhwani SA, Chopade B. Bacteriagenic silver nanoparticles: synthesis, mechanism, and applications. Appl Microbiol Biotechnol 2015; 99: 4579-4593.
13. Bogutska K, Sklyarov YP, Prylutskyy Y. Zinc and zinc nanoparticles: biological role and application in biomedicine. Ukr Bioorg Acta 2013; 1: 9-16.
14. Eivazihollagh A, Bäckström J, Dahlström C, Carlsson F, Ibrahem I, Lindman B, et al. One-pot synthesis of cellulose-templated copper nanoparticles with antibacterial properties. Mat Lett 2017; 187: 170-172.
15. Chatterjee AK, Chakraborty R, Basu T. Mechanism of antibacterial activity of copper nanoparticles. Nanotechnology 2014; 25: 135101.
16. Nabila MI, Kannabiran K. Biosynthesis, characterization and antibacterial activity of copper oxide nanoparticles (CuO NPs) from actinomycetes. Biocatal Agric Biotechnol 2018; 15: 56-62.
17. Shahmohamadi S, Khosravi M. Effect of hydroalcoholic extract of Salvia officinalis L. on the activity of catalase and superoxide dismutase in an oxidative stress model created by intracerebroventricular STZ injection in male rats. Physiol Pharmacol 2013; 17: 176-184.
18. Alnaddaf LM, Almuhammady AK, Salem KF, Alloosh MT, Saleh MM, Al-Khayri JM (2021). Green synthesis of nanoparticles using different plant extracts and their characterizations. In: Nanobiotechnology: Mitigation of Abiotic Stress in Plants. Cham: Springer; pp. 165-199.
19. Kaviya S, Santhanalakshmi J, Viswanathan B, Muthumary J, Srinivasan K. Biosynthesis of silver nanoparticles using Citrus sinensis peel extract and its antibacterial activity. Spectrochim Acta A Mol Biomol Spectrosc 2011; 79: 594-598.
20. Zangeneh MM, Joshani Z, Zangeneh A, Miri E. Green synthesis of silver nanoparticles using aqueous extract of Stachys lavandulifolia flower, and their cytotoxicity, antioxidant, antibacterial and cutaneous wound‐healing properties. Appl Organomet Chem 2019; 33(9): e5016.
21. Govindaraju K, Kiruthiga V, Kumar VG, Singaravelu G. Extracellular synthesis of silver nanoparticles by a marine alga, Sargassum wightii Grevilli and their antibacterial effects. J Nanosci Nanotechnol 2009; 9: 5497-5501.
22. Mirzaei SZ, Lashgarian HE, Karkhane M, Shahzamani K, Alhameedawi AK, Marzban A, et al. Bio-inspired silver selenide nano-chalcogens using aqueous extract of Melilotus officinalis with biological activities. Bioresour Bioprocess 2021; 8: 56.
23. Hosseinimehr SJ, Mahmoudzadeh A, Ahmadi A, Ashrafi SA, Shafaghati N, Hedayati N. The radioprotective effect of Zataria multiflora against genotoxicity induced by γ irradiation in human blood lymphocytes. Cancer Biother Radiopharm 2011; 26: 325-329.
24. Gawande MB, Goswami A, Felpin FX, Asefa T, Huang X, Silva R, et al. Cu and Cu-based nanoparticles: synthesis and applications in catalysis. Chem Rev 2016; 116: 3722-3811.
25. Naika HR, Lingaraju K, Manjunath K, Kumar D, Nagaraju G, Suresh D, et al. Green synthesis of CuO nanoparticles using Gloriosa superba L. extract and their antibacterial activity. J Taibah Univ Sci 2015; 9: 7-12.
26. Waris A, Din M, Ali A, Ali M, Afridi S, Baset A, et al. A comprehensive review of green synthesis of copper oxide nanoparticles and their diverse biomedical applications. Inorg Chem Commun 2021; 123: 108369.
27. Murugan B, Rahman MZ, Fatimah I, Lett JA, Annaraj J, Kaus NH, et al. Green synthesis of CuO nanoparticles for biological applications. Inorg Chem Commun 2023; 155: 111088.
28. Nethravathi P, Kumar MP, Suresh D, Lingaraju K, Rajanaika H, Nagabhushana H, et al. Tinospora cordifolia mediated facile green synthesis of cupric oxide nanoparticles and their photocatalytic, antioxidant and antibacterial properties. Mater Sci Semicond Process 2015; 33: 81-88.
29. Saif S, Tahir A, Asim T, Chen Y. Plant mediated green synthesis of CuO nanoparticles: comparison of toxicity of engineered and plant mediated CuO nanoparticles towards Daphnia magna. Nanomaterials (Basel) 2016; 6: 205.
30. Noor S, Shah Z, Javed A, Ali A, Hussain SB, Zafar S, et al. A fungal based synthesis method for copper nanoparticles with the determination of anticancer, antidiabetic and antibacterial activities. J Microbiol Methods 2020; 174: 105966.
31. Sathiyavimal S, Vasantharaj S, Veeramani V, Saravanan M, Rajalakshmi G, Kaliannan T, et al. Green chemistry route of biosynthesized copper oxide nanoparticles using Psidium guajava leaf extract and their antibacterial activity and effective removal of industrial dyes. J Environ Chem Eng 2021; 9: 105033.
32. Khairy T, Amin DH, Salama HM, Elkholy IMA, Elnakib M, Gebreel HM, et al. Antibacterial activity of green synthesized copper oxide nanoparticles against multidrug-resistant bacteria. Sci Rep 2024; 14: 25020.
33. Chandrakanth RK, Ashajyothi C, Oli AK, Prabhurajeshwar C. Potential bactericidal effect of silver nanoparticles synthesised from Enterococcus species. Orient J Chem 2014; 30: 3.
34. Das PE, Abu-Yousef IA, Majdalawieh AF, Narasimhan S, Poltronieri P. Green synthesis of encapsulated copper nanoparticles using a hydroalcoholic extract of Moringa oleifera leaves and assessment of their antioxidant and antimicrobial activities. Molecules 2020; 25: 555.
35. Rajivgandhi G, Maruthupandy M, Muneeswaran T, Ramachandran G, Manoharan N, Quero F, et al. Biologically synthesized copper oxide nanoparticles enhanced intracellular damage in ciprofloxacin resistant ESBL producing bacteria. Microb Pathog 2019; 127: 267-276.
36. Prabhu Y, Rao KV, Sai VS, Pavani T. A facile biosynthesis of copper nanoparticles: A micro-structural and antibacterial activity investigation. J Saudi Chem Soc 2017; 21: 180-185.
37. Agarwala M, Choudhury B, Yadav R. Comparative study of antibiofilm activity of copper oxide and iron oxide nanoparticles against multidrug resistant biofilm forming uropathogens. Indian J Microbiol 2014; 54: 365-368.
38. Mohanta YK, Biswas K, Jena SK, Hashem A, Abd Allah EF, Mohanta TK. Anti-biofilm and antibacterial activities of silver nanoparticles synthesized by the reducing activity of phytoconstituents present in the Indian medicinal plants. Front Microbiol 2020; 11: 1143.
39. Shakib P, Mirzaei SZ, Sharafi Z, Saki R, Goudarzi GR, Sepeavand A, et al. Biofabrication of copper oxide nanoparticles mediated with Echium amoenum petal extract for evaluation of biological functions. Biomass Convers Biorefin 2024; 14: 25651-25661.
40. Baharara J, Ramezani T, Mousavi M, Asadi-Samani M. Antioxidant and anti-inflammatory activity of green synthesized silver nanoparticles using Salvia officinalis extract. Ann Trop Med Public Health 2017; 10: 1265-1270.
41. Zamanian Z, Tajbakhsh E, Arbab Soleimani N, Ghasemian A. Aqueous extract‐mediated green synthesis of CuO nanoparticles: Potential anti‐tuberculosis agents. Food Sci Nutr 2024; 12: 5907-5921.
42. Barani M, Mir A, Roostaee M, Sargazi G, Adeli-Sardou M. Green synthesis of copper oxide nanoparticles via Moringa peregrina extract incorporated in graphene oxide: evaluation of antibacterial and anticancer efficacy. Bioprocess Biosyst Eng 2024; 47: 1915-1928.
43. Ranjbar M, Khakdan F, Mukherjee A. In vitro analysis of green synthesized CuO nanoparticles using Tanacetum parthenium extract for multifunctional applications. Environ Sci Pollut Res Int 2023; 30: 60180-60195.
2. Saxena A, Tripathi R, Zafar F, Singh P. Green synthesis of silver nanoparticles using aqueous solution of Ficus benghalensis leaf extract and characterization of their antibacterial activity. Mater Lett 2012; 67: 91-94.
3. Gour A, Jain NK. Advances in green synthesis of nanoparticles. Artif Cells Nanomed Biotechnol 2019; 47: 844-851.
4. Hussain I, Singh N, Singh A, Singh H, Singh S. Green synthesis of nanoparticles and its potential application. Biotechnol Lett 2016; 38: 545-560.
5. Villagrán Z, Anaya-Esparza LM, Velázquez-Carriles CA, Silva-Jara JM, Ruvalcaba-Gómez JM, Aurora-Vigo EF, et al. Plant-based extracts as reducing, capping, and stabilizing agents for the green synthesis of inorganic nanoparticles. Resources 2024; 13: 70.
6. Jadoun S, Arif R, Jangid NK, Meena R. Green synthesis of nanoparticles using plant extracts: A review. Environ Chem Lett 2021; 19: 355-374.
7. Nadaroglu H, GÜNGÖR AA, Selvi İ. Synthesis of nanoparticles by green synthesis method. Int J Innov Res Rev 2017; 1: 6-9.
8. Grdiša M, Jug-Dujaković M, Lončarić M, Carović-Stanko K, Ninčević T, Liber Z, et al. Dalmatian sage (Salvia officinalis L.): A review of biochemical contents, medical properties and genetic diversity. Agric Conspec Sci 2015; 80: 69-78.
9. Lu Y, Foo LY. Polyphenolics of Salvia—a review. Phytochemistry 2002 ; 59: 117-140.
10. Walch SG, Kuballa T, Stühlinger W, Lachenmeier D. Determination of the biologically active flavour substances thujone and camphor in foods and medicines containing sage (Salvia officinalis L.). Chem Cent J 2011; 5: 44.
11. Khakpour S, Khosravi M, Jafari Marandi S, Ahadi Ali M. The effect of hydroalcoholic extract of Salvia officinalis L. on the inflammatory reduction in male mice. AMU-TMUJ 2014; 24: 136-142.
12. Singh R, Shedbalkar UU, Wadhwani SA, Chopade B. Bacteriagenic silver nanoparticles: synthesis, mechanism, and applications. Appl Microbiol Biotechnol 2015; 99: 4579-4593.
13. Bogutska K, Sklyarov YP, Prylutskyy Y. Zinc and zinc nanoparticles: biological role and application in biomedicine. Ukr Bioorg Acta 2013; 1: 9-16.
14. Eivazihollagh A, Bäckström J, Dahlström C, Carlsson F, Ibrahem I, Lindman B, et al. One-pot synthesis of cellulose-templated copper nanoparticles with antibacterial properties. Mat Lett 2017; 187: 170-172.
15. Chatterjee AK, Chakraborty R, Basu T. Mechanism of antibacterial activity of copper nanoparticles. Nanotechnology 2014; 25: 135101.
16. Nabila MI, Kannabiran K. Biosynthesis, characterization and antibacterial activity of copper oxide nanoparticles (CuO NPs) from actinomycetes. Biocatal Agric Biotechnol 2018; 15: 56-62.
17. Shahmohamadi S, Khosravi M. Effect of hydroalcoholic extract of Salvia officinalis L. on the activity of catalase and superoxide dismutase in an oxidative stress model created by intracerebroventricular STZ injection in male rats. Physiol Pharmacol 2013; 17: 176-184.
18. Alnaddaf LM, Almuhammady AK, Salem KF, Alloosh MT, Saleh MM, Al-Khayri JM (2021). Green synthesis of nanoparticles using different plant extracts and their characterizations. In: Nanobiotechnology: Mitigation of Abiotic Stress in Plants. Cham: Springer; pp. 165-199.
19. Kaviya S, Santhanalakshmi J, Viswanathan B, Muthumary J, Srinivasan K. Biosynthesis of silver nanoparticles using Citrus sinensis peel extract and its antibacterial activity. Spectrochim Acta A Mol Biomol Spectrosc 2011; 79: 594-598.
20. Zangeneh MM, Joshani Z, Zangeneh A, Miri E. Green synthesis of silver nanoparticles using aqueous extract of Stachys lavandulifolia flower, and their cytotoxicity, antioxidant, antibacterial and cutaneous wound‐healing properties. Appl Organomet Chem 2019; 33(9): e5016.
21. Govindaraju K, Kiruthiga V, Kumar VG, Singaravelu G. Extracellular synthesis of silver nanoparticles by a marine alga, Sargassum wightii Grevilli and their antibacterial effects. J Nanosci Nanotechnol 2009; 9: 5497-5501.
22. Mirzaei SZ, Lashgarian HE, Karkhane M, Shahzamani K, Alhameedawi AK, Marzban A, et al. Bio-inspired silver selenide nano-chalcogens using aqueous extract of Melilotus officinalis with biological activities. Bioresour Bioprocess 2021; 8: 56.
23. Hosseinimehr SJ, Mahmoudzadeh A, Ahmadi A, Ashrafi SA, Shafaghati N, Hedayati N. The radioprotective effect of Zataria multiflora against genotoxicity induced by γ irradiation in human blood lymphocytes. Cancer Biother Radiopharm 2011; 26: 325-329.
24. Gawande MB, Goswami A, Felpin FX, Asefa T, Huang X, Silva R, et al. Cu and Cu-based nanoparticles: synthesis and applications in catalysis. Chem Rev 2016; 116: 3722-3811.
25. Naika HR, Lingaraju K, Manjunath K, Kumar D, Nagaraju G, Suresh D, et al. Green synthesis of CuO nanoparticles using Gloriosa superba L. extract and their antibacterial activity. J Taibah Univ Sci 2015; 9: 7-12.
26. Waris A, Din M, Ali A, Ali M, Afridi S, Baset A, et al. A comprehensive review of green synthesis of copper oxide nanoparticles and their diverse biomedical applications. Inorg Chem Commun 2021; 123: 108369.
27. Murugan B, Rahman MZ, Fatimah I, Lett JA, Annaraj J, Kaus NH, et al. Green synthesis of CuO nanoparticles for biological applications. Inorg Chem Commun 2023; 155: 111088.
28. Nethravathi P, Kumar MP, Suresh D, Lingaraju K, Rajanaika H, Nagabhushana H, et al. Tinospora cordifolia mediated facile green synthesis of cupric oxide nanoparticles and their photocatalytic, antioxidant and antibacterial properties. Mater Sci Semicond Process 2015; 33: 81-88.
29. Saif S, Tahir A, Asim T, Chen Y. Plant mediated green synthesis of CuO nanoparticles: comparison of toxicity of engineered and plant mediated CuO nanoparticles towards Daphnia magna. Nanomaterials (Basel) 2016; 6: 205.
30. Noor S, Shah Z, Javed A, Ali A, Hussain SB, Zafar S, et al. A fungal based synthesis method for copper nanoparticles with the determination of anticancer, antidiabetic and antibacterial activities. J Microbiol Methods 2020; 174: 105966.
31. Sathiyavimal S, Vasantharaj S, Veeramani V, Saravanan M, Rajalakshmi G, Kaliannan T, et al. Green chemistry route of biosynthesized copper oxide nanoparticles using Psidium guajava leaf extract and their antibacterial activity and effective removal of industrial dyes. J Environ Chem Eng 2021; 9: 105033.
32. Khairy T, Amin DH, Salama HM, Elkholy IMA, Elnakib M, Gebreel HM, et al. Antibacterial activity of green synthesized copper oxide nanoparticles against multidrug-resistant bacteria. Sci Rep 2024; 14: 25020.
33. Chandrakanth RK, Ashajyothi C, Oli AK, Prabhurajeshwar C. Potential bactericidal effect of silver nanoparticles synthesised from Enterococcus species. Orient J Chem 2014; 30: 3.
34. Das PE, Abu-Yousef IA, Majdalawieh AF, Narasimhan S, Poltronieri P. Green synthesis of encapsulated copper nanoparticles using a hydroalcoholic extract of Moringa oleifera leaves and assessment of their antioxidant and antimicrobial activities. Molecules 2020; 25: 555.
35. Rajivgandhi G, Maruthupandy M, Muneeswaran T, Ramachandran G, Manoharan N, Quero F, et al. Biologically synthesized copper oxide nanoparticles enhanced intracellular damage in ciprofloxacin resistant ESBL producing bacteria. Microb Pathog 2019; 127: 267-276.
36. Prabhu Y, Rao KV, Sai VS, Pavani T. A facile biosynthesis of copper nanoparticles: A micro-structural and antibacterial activity investigation. J Saudi Chem Soc 2017; 21: 180-185.
37. Agarwala M, Choudhury B, Yadav R. Comparative study of antibiofilm activity of copper oxide and iron oxide nanoparticles against multidrug resistant biofilm forming uropathogens. Indian J Microbiol 2014; 54: 365-368.
38. Mohanta YK, Biswas K, Jena SK, Hashem A, Abd Allah EF, Mohanta TK. Anti-biofilm and antibacterial activities of silver nanoparticles synthesized by the reducing activity of phytoconstituents present in the Indian medicinal plants. Front Microbiol 2020; 11: 1143.
39. Shakib P, Mirzaei SZ, Sharafi Z, Saki R, Goudarzi GR, Sepeavand A, et al. Biofabrication of copper oxide nanoparticles mediated with Echium amoenum petal extract for evaluation of biological functions. Biomass Convers Biorefin 2024; 14: 25651-25661.
40. Baharara J, Ramezani T, Mousavi M, Asadi-Samani M. Antioxidant and anti-inflammatory activity of green synthesized silver nanoparticles using Salvia officinalis extract. Ann Trop Med Public Health 2017; 10: 1265-1270.
41. Zamanian Z, Tajbakhsh E, Arbab Soleimani N, Ghasemian A. Aqueous extract‐mediated green synthesis of CuO nanoparticles: Potential anti‐tuberculosis agents. Food Sci Nutr 2024; 12: 5907-5921.
42. Barani M, Mir A, Roostaee M, Sargazi G, Adeli-Sardou M. Green synthesis of copper oxide nanoparticles via Moringa peregrina extract incorporated in graphene oxide: evaluation of antibacterial and anticancer efficacy. Bioprocess Biosyst Eng 2024; 47: 1915-1928.
43. Ranjbar M, Khakdan F, Mukherjee A. In vitro analysis of green synthesized CuO nanoparticles using Tanacetum parthenium extract for multifunctional applications. Environ Sci Pollut Res Int 2023; 30: 60180-60195.
| Files | ||
| Issue | Vol 18 No 3 (2026) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v18i3.21660 | |
| Keywords | ||
| Green synthesis Copper oxide nanoparticles (CuO-NPs) Antimicrobial Antioxidant Salvia officinalis | ||
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How to Cite
1.
Marzban A, Shakib P, Mirzaei SZ, Goudarzi G, Jahani F. Antimicrobial and antioxidant properties of green synthesis CuO nanoparticles using Salvia officinalis hydroalcoholic extract. Iran J Microbiol. 2026;18(3):345-356.
