Green synthesis of mixed-phase copper nanostructures using Annona muricata extract: role of oxidation on selective antifungal activity
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Keywords:
Green synthesis, copper nanoparticles, Annona muricata, Phytochemical reduction, Antimicrobial activity
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Abstract
The green synthesis of copper-based nanomaterials utilizing plant extracts has attracted significant attention; however, controlling oxidation state and understanding its impact on biological performance remain challenging. In this present study, mixed-phase copper nanostructures were synthesized using aqueous stem bark extract of Annona muricata as a dual reducing and stabilizing agent under mild conditions. Phytochemical analysis revealed a high total phenolic content (310 mg GAE/g), thus confirming the strong redox capability for Cu2+ reduction. UV–Vis spectroscopy indicated the formation of nanoparticle with a characteristic absorption band at approximately 360 nm. XRD and EDS analyses were conducted to confirm the coexistence of Cu (0), Cu2O, and CuO phases. The crystallite sizes of these phases ranged from 22 to 28 nm, thereby suggesting partial oxidation during synthesis. The utilization of Fourier-transform infrared spectroscopy (FTIR) analysis has been demonstrated to be a valuable tool in the identification of the involvement of polyphenolic functional groups in reduction and surface stabilization processes. SEM and TEM observations revealed the presence of quasi-spherical nanoparticles with a mean diameter of 4 -16 nm, exhibiting signs of partial aggregation. The antimicrobial performance of the synthesized nanostructures exhibited a concentration-dependent response. While limited antibacterial activity was observed against Staphylococcus aureus and Escherichia coli, a pronounced antifungal effect was obtained against Candida albicans with an inhibition zone of 27.81 mm at 20% concentration. The pronounced antifungal activity exhibited a strong correlation with the presence of mixed copper phases, suggesting that the oxidation-induced surface chemistry and controlled Cu2+ ion availability may contribute significantly to the observed efficacy. These findings highlight the functional role of oxidation in plant-mediated copper nanostructures and demonstrate their potential as selective antifungal agents. This work provides insight into the relationship between phase composition and biological activity in green-synthesized copper nanomaterials. The remarkable selectivity exhibited by these mixed-phase nanostructures against fungal pathogens positions them as a viable, eco-friendly alternative for targeted antifungal applications, thus overcoming the limitations of conventional non-specific antimicrobial agents.
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Nurjanah, Mustafa, I., Ulfika, T., Ginting, B., & Maulana, I. (2026). Green synthesis of mixed-phase copper nanostructures using Annona muricata extract: role of oxidation on selective antifungal activity. Communications in Science and Technology, 11(1), 169–179. https://doi.org/10.21924/cst.11.1.2026.1967
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References
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2. B.D. Harishchandra, M. Pappuswamy, G. Shama, V.A. Arumugam, T. Periyaswamy, R. Sundaram, Copper nanoparticles: a review on synthesis, characterization and applications. Asian Pacific Journal of Cancer Biology 5 (2020) 201–210.
3. S. Sampaio, J.C. Viana, Optimisation of the green synthesis of Cu/Cu2O particles for maximum yield production and reduced oxidation for electronic applications. Materials Science and Engineering: B 263 (2021).
4. J. Leitner, D. Sedmidubský, M. Lojka, O. Jankovský, The effect of nanosizing on the oxidation of partially oxidized copper nanoparticles. Materials 13 (2020).
5. Z.X. Chen, Y.J. Song, R.Z. Li, S.J. Guo, L. Shi, Z.R. Yang, X.M. Xue, T. Zhang, Advances in copper nanocrystals: Synthesis, anti-oxidation strategies, and multiple applications. Coord. Chem. Rev. 529 (2025).
6. A.I. Osman, Y. Zhang, M. Farghali, A.K. Rashwan, A.S. Eltaweil, E.M. Abd El-Monaem, I.M.A. Mohamed, M.M. Badr, I. Ihara, D.W. Rooney, P.S. Yap, Synthesis of green nanoparticles for energy, biomedical, environmental, agricultural, and food applications: A review. Environ. Chem. Lett. 22 (2024) 841–887.
7. H. Singh, M.F. Desimone, S. Pandya, S. Jasani, N. George, M. Adnan, A. Aldarhami, A.S. Bazaid, S.A. Alderhami, Revisiting the Green Synthesis of Nanoparticles: Uncovering Influences of Plant Extracts as Reducing Agents for Enhanced Synthesis Efficiency and Its Biomedical Applications. Int. J. Nanomedicine 18 (2023) 4727–4750.
8. M. Bin Mobarak, M.F. Sikder, K.S. Muntaha, S. Islam, S.M.F. Rabbi, F. Chowdhury, Plant extract-mediated green-synthesized CuO nanoparticles for environmental and microbial remediation: a review covering basic understandings to mechanistic study. Nanoscale Adv. 7 (2025) 2418–2445.
9. A. Dhaka, S. Chand Mali, S. Sharma, R. Trivedi, A review on biological synthesis of silver nanoparticles and their potential applications. Results Chem. 6 (2023).
10. H.R. El-Seedi, M.S. Omara, A.H. Omar, M.M. Elakshar, Y.M. Shoukhba, H. Duman, S. Karav, A.K. Rashwan, A.H. El-Seedi, H.A. Altaleb, H. Gao, A. Saeed, O.A. Jefri, Z. Guo, S.A.M. Khalifa, Updated Review of Metal Nanoparticles Fabricated by Green Chemistry Using Natural Extracts: Biosynthesis, Mechanisms, and Applications. Bioengineering 11 (2024).
11. S. Iravani, Green synthesis of metal nanoparticles using plants. Green Chemistry 13 (2011) 2638–2650.
12. I.L. Santos, A.M. da C. Rodrigues, E.R. Amante, L.H.M. da Silva, Soursop (Annona muricata) Properties and Perspectives for Integral Valorization. Foods 12 (2023).
13. M. Mutakin, R. Fauziati, F.N. Fadhilah, A. Zuhrotun, R. Amalia, Y.E. Hadisaputri, Pharmacological Activities of Soursop (Annona muricata Lin.). Molecules 27 (2022).
14. D. Letchumanan, S.P.M. Sok, S. Ibrahim, N.H. Nagoor, N.M. Arshad, Plant-based biosynthesis of copper/copper oxide nanoparticles: An update on their applications in biomedicine, mechanisms, and toxicity. Biomolecules 11 (2021).
15. S. Kayalvizhi, A. Sengottaiyan, T. Selvankumar, B. Senthilkumar, C. Sudhakar, K. Selvam, Eco-friendly cost-effective approach for synthesis of copper oxide nanoparticles for enhanced photocatalytic performance. Optik (Stuttg). 202 (2020).
16. S. Amaliyah, D.P. Pangesti, M. Masruri, A. Sabarudin, S.B. Sumitro, Green synthesis and characterization of copper nanoparticles using Piper retrofractum Vahl extract as bioreductor and capping agent. Heliyon 6 (2020).
17. E. Takele Assefa, G. Shumi, K. Mohammed Gendo, G. Kenasa, N. Roba, Review on green synthesis, characterization, and antibacterial activity of CuO nanoparticles using biomolecules of plant extract. Results Chem. 8 (2024).
18. X. Ma, S. Zhou, X. Xu, Q. Du, Copper-containing nanoparticles: Mechanism of antimicrobial effect and application in dentistry-a narrative review. Front. Surg. 9 (2022).
19. A.R. Noviyanti, E.N. Asyiah, M.D. Permana, D. Dwiyanti, Suryana, D.R. Eddy, Preparation of Hydroxyapatite-Titanium Dioxide Composite from Eggshell by Hydrothermal Method: Characterization and Antibacterial Activity. Crystals (Basel). 12 (2022).
20. L.K. Sakti, G.A.N. Sheha, C.P.S.M. Radhiyah, J. Nugraha, D.R. Eddy, M.D. Permana, I.P. Maksum, Y. Deawati, Cotton Fabric Coating by rGO and Polymethylsiloxane Layer with Antibacterial, Hydrophobic and Photothermal Properties. Trends in Sciences 20 (2023).
21. J.Y. Park, S. Lee, Y. Kim, Y.B. Ryu, Antimicrobial Activity of Morphology-Controlled Cu2O Nanoparticles: Oxidation Stability under Humid and Thermal Conditions. Materials 17 (2024).
22. L. Wang, C. Hu, L. Shao, The antimicrobial activity of nanoparticles: Present situation and prospects for the future. Int. J. Nanomedicine 12 (2017) 1227–1249.
23. K. Madeshwaran, R. Venkatachalam, Green synthesis of bimetallic ZnO–CuO nanoparticles using Annona muricata l. extract: Investigation of antimicrobial, antioxidant, and anticancer properties. Journal of Industrial and Engineering Chemistry 140 (2024) 454–467.
24. R.I. Mahmood, A.A. Kadhim, S. Ibraheem, S. Albukhaty, H.S. Mohammed-Salih, R.H. Abbas, M.S. Jabir, M.K.A. Mohammed, U.M. Nayef, F.A. AlMalki, G.M. Sulaiman, H. Al-Karagoly, Biosynthesis of copper oxide nanoparticles mediated Annona muricata as cytotoxic and apoptosis inducer factor in breast cancer cell lines. Sci. Rep. 12 (2022).
25. I. Maulana, B. Ginting, K. Azizah, Green synthesis of copper nanoparticles employing Annona squamosa L extract as antimicrobial and anticancer agents. S. Afr. J. Chem. Eng. 46 (2023) 65–71.
26. I. Maulana, D. Fasya, B. Ginting, Biosynthesis of Cu nanoparticles using Polyalthia longifolia roots extracts for antibacterial, antioxidant and cytotoxicity applications. Materials Technology 37 (2022) 2517–2521.
27. G.I. Edo, A.N. Mafe, A.B.M. Ali, P.O. Akpoghelie, E. Yousif, E.F. Isoje, U.A. Igbuku, K. Zainulabdeen, J.O. Owheruo, A.E.A. Essaghah, H. Umar, D.S. Ahmed, A.A. Alamiery, Eco-friendly nanoparticle phytosynthesis via plant extracts: Mechanistic insights, recent advances, and multifaceted uses. Nano TransMed 4 (2025).
28. A.N. Labaran, Z.U. Zango, G. Tailor, A. Alsadig, F. Usman, M.T. Mukhtar, A.M. Garba, R. Alhathlool, K.H. Ibnaouf, O.A. Aldaghri, Biosynthesis of copper nanoparticles using Alstonia scholaris leaves and its antimicrobial studies. Sci. Rep. 14 (2024).
29. R. Amjad, B. Mubeen, S.S. Ali, S.S. Imam, S. Alshehri, M.M. Ghoneim, S.I. Alzarea, R. Rasool, I. Ullah, M.S. Nadeem, I. Kazmi, Green synthesis and characterization of copper nanoparticles using fortunella margarita leaves. Polymers (Basel). 13 (2021).
30. H.T. Gebrie, M.A. Assege, D.S. Meshesha, B.A. Tebeje, G.W. Moges, A.T. Wodajo, G.M. Manahelohe, A.A. Belew, Biosynthesis and characterization of copper oxide nanoparticles from Plumbago zeylanica leaf extract for antibacterial and antioxidant activities. Sci. Rep. 15 (2025).
31. M. V. Faustino, M.A.F. Faustino, H. Silva, Â. Cunha, A.M.S. Silva, D.C.G.A. Pinto, Puccinellia maritima, Spartina maritime, and Spartina patens Halophytic Grasses: Characterization of polyphenolic and chlorophyll profiles and evaluation of their biological activities. Molecules 24 (2019).
32. Y. Yu, L. Zhang, J. Wang, Z. Yang, M. Long, N. Hu, Y. Zhang, Preparation of hollow porous Cu2O microspheres and photocatalytic activity under visible light irradiation. Nanoscale Res. Lett. 7 (2012).
33. N. Munandar, H.F. Aritonang, R. Bonaventura, D.P. Wijaya, Synthesis of Cu Nanoparticles using Anredera cordifolia Extract and their Potential as Antidiabetic with Alpha Amylase Enzyme Inhibition. Communications in Science and Technology 10 (2025) 422–430.
34. S. Nareshkumar, K. Madhivanan, A.K. Sundramoorthy, Development of a hybrid nanomaterial-based electrochemical sensor for the determination of L-tyrosine using copper oxide and SWCNTs. Journal of SolidState Electrochemistry (2026).
35. E.A. Mohamed, Green synthesis of copper & copper oxide nanoparticles using the extract of seedless dates. Heliyon 6 (2020).
36. H.M. Albert, C.A. Gonsago, Green Procedure for the synthesis of Copper Nanoparticles using Nerium oleander Leaf Extract: Characterizations and Applications. Oriental Journal Of Chemistry 39 (2023) 792–797.
37. akl awwad and mohammad amer, Green Synthesis of Copper Nanoparticles by Citrus Limon Fruits Extract, Characterization and Antibacterial Activity. International Scientific Organization 7 (1) (2021) 1-8.
38. H. Koohestani, A. Balooch, H. Staji, Biosynthesis of CuO-nanoparticles using extracts of Mespilus germanica, Crataegus spp., and Wild barberry and their synergistic antibacterial activity. Giant 26 (2025).
39. M.J. Javid-Naderi, Z. Sabouri, A. Jalili, H. Zarrinfar, S. Sammak, M. Darroudi, Green synthesis and characterization of Ag/CuO nanoparticles: Exploring their antifungal, antimicrobial, and cytotoxic properties. Environ. Technol. Innov. 38 (2025).
40. M.D. Moroda, T. Leta Deressa, A.H. Tiwikrama, T.F. Chala, Green synthesis of copper oxide nanoparticles using Rosmarinus officinalis leaf extract and evaluation of its antimicrobial activity. Next Materials 7 (2025).
41. I. Maulana, B. Ginting, I. Mustafa, R.A.N. Islami, Green Synthesis of Copper Nanoparticles Using Polyalthia longifolia Roots and their Bioactivities Against Escherichia coli, Staphylococcus aureus, and Candida albicans. J. Pharm. Bioallied Sci. 16 (2024) S2218–S2223.