Green Synthesis and Characterization of Zinc Oxide Nanoparticles Using Hedysarum Varium Wild. and Its Antibacterial Activities

Year 2025, Volume: 20 Issue: 2, 194 - 205, 26.11.2025

Ece Ozkan , Batuhan Ozturk , Gizem Arik , Fatma Taeb Dişli , Mehmet Bay , Suzan Biran Ay , Nihan Kosku Perkgöz , İsmail Murat Palabıyık

Abstract

The increasing interest in nanomaterials has accelerated the search for sustainable and eco-friendly synthesis routes. In this study, zinc oxide nanoparticles (ZnO NPs) were successfully synthesized for the first time using Hedysarum varium extracts through a green synthesis approach. The plant material was extracted using a methanol–water mixture, and the resulting extract was incorporated into a zinc nitrate solution, followed by incubation at 60 °C for 4 hours under alkaline conditions (pH 10). After purification and drying at 90 °C, pure ZnO NPs were obtained. Structural and morphological properties were characterized by UV-Vis spectroscopy, optical microscopy, Raman spectroscopy, XRD, and atomic force microscopy (AFM), confirming the formation of crystalline ZnO NPs with average sizes ranging from 10–20 nm, while optimization studies revealed that a 25 mg extract concentration provided the most stable and controlled nanoparticle formation. Antibacterial activity was evaluated against Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Staphylococcus aureus ATCC 25923, and the nanoparticles exhibited the strongest effect against Gram-positive S. aureus (MIC: 0.25 mg/mL), indicating potential for biomedical applications. Compared with previously reported green synthesis methods using Aloe vera or Thymbra spicata, the use of H. varium extract offers a novel, cost-effective, and sustainable alternative with antibacterial properties. This study contributes to the literature by presenting the first report of ZnO NP synthesis mediated by H. varium, highlighting its promising role as a green source for nanomaterial production, and suggesting that future studies should expand microbial spectrum testing, explore cytotoxicity and biocompatibility, and evaluate potential industrial-scale applications.

Keywords

Green synthesis , Hedysarum varium , Zinc oxide , Nanoparticles , Plants extract

Ethical Statement

No human or animal subjects were used in this study.

Thanks

Ece Özkan thanks the financial support from the Scientific and Technological Research Council of Türkiye (TUBITAK) under the BIDEB/ 2218 National Postdoctoral Research Scholarship Program (project no. 122C249). Batuhan Öztürk thanks the financial support from the Scientific and Technological Research Council of Türkiye (TUBITAK) under the BIDEB/ 2210 MSc/MA Scholarship Programs.

References

• Y. Cao, B. Liu, Z. Xia, S. Ge, and R. Huang, "Flash synthesis of flower-like ZnO nanostructures by microwave-induced combustion process," (in English), Materials Letters, vol. 65, no. 2, pp. 160-163, 2011, doi: 10.1016/j.matlet.2010.09.072.
• F. Shabnam, J. Mina, and F. Hassan Kabiri, "Green synthesis of zinc oxide nanoparticles: a comparison," in Green Chemistry Letters and Reviews vol. 12, ed: Taylor & Francis Group, 2019, pp. 19-24.
• H. Abdul Salam, R. Sivaraj, and R. Venckatesh, "Green synthesis and characterization of zinc oxide nanoparticles from Ocimum basilicum L. var. purpurascens Benth.-Lamiaceae leaf extract," (in English), Materials Letters, vol. 131, pp. 16-18, 2014, doi: 10.1016/j.matlet.2014.05.033.
• R. Yuvakkumar, J. Suresh, A. J. Nathanael, M. Sundrarajan, and S. I. Hong, "Novel green synthetic strategy to prepare ZnO nanocrystals using rambutan (Nephelium lappaceum L.) peel extract and its antibacterial applications," (in English), Materials Science & Engineering C, vol. 41, pp. 17-27, 2014, doi: 10.1016/j.msec.2014.04.025.
• H. Agarwal, S. Venkat Kumar, and S. Rajeshkumar, "A review on green synthesis of zinc oxide nanoparticles – An eco-friendly approach," (in English), Resource-efficient technologies, vol. 3, no. 4, pp. 406-413, 2017, doi: 10.1016/j.reffit.2017.03.002.
• E. Boissier and R. Buser, Flora Orientalis : sive, Enumeratio plantarum in Oriente a Graecia et Aegypto ad Indiae fines hucusque observatarum. Basileae: H. Georg, 1867, p. 31.
• "Legumes of Northern Eurasia. A Checklist G. P. Yakovlev A. K. Sytin Yu. R. Roskov," in Kew Bulletin vol. 51, ed, 1996, pp. 622-623.
• H. Hoşgören and A. S. Ertekin, "A new record for the Flora of Turkey, Hedysarum singarense Boiss. & Hausskn. (Fabaceae)," (in en), International Journal of Nature and Life Sciences, vol. 2, no. 1, pp. 10-16, June 2018. [Online]. Available: https://dergipark.org.tr/en/pub/ijnls/issue/43939/541103.
• T. Ertugrul, B. Yildiz, M. G. Ozturk, A. Fisne, E. Bag, and Z. Aytac, "Hedysarum vuralii (Fabaceae), a new species from Turkey," (in English), PHYTOTAXA, vol. 641, no. 1, pp. 47-56, 2024, doi: 10.11646/phytotaxa.641.1.4.
• B.-H. Choi and H. Ohashi, "Generic Criteria and an Infrageneric System for Hedysarum and Related Genera (Papilionoideae-Leguminosae)," (in English), Taxon, vol. 52, no. 3, pp. 567-576, 2003, doi: 10.2307/3647455.
• G. Zengin, G. O. Guler, A. Aktumsek, R. Ceylan, C. M. N. Picot, and M. F. Mahomoodally, "Enzyme Inhibitory Properties, Antioxidant Activities, and Phytochemical Profile of Three Medicinal Plants from Turkey," (in eng), Advances in Pharmacological Sciences, pp. 1-8, 2015, doi: 10.1155/2015/410675.
• J. Santhoshkumar, S. V. Kumar, and S. Rajeshkumar, "Synthesis of zinc oxide nanoparticles using plant leaf extract against urinary tract infection pathogen," (in English), Resource-efficient technologies, vol. 3, no. 4, pp. 459-465, 2017, doi: 10.1016/j.reffit.2017.05.001.
• G. Sangeetha, S. Rajeshwari, and R. Venckatesh, "Green synthesis of zinc oxide nanoparticles by aloe barbadensis miller leaf extract: Structure and optical properties," (in English), Materials Research Bulletin, vol. 46, no. 12, pp. 2560-2566, 2011, doi: 10.1016/j.materresbull.2011.07.046.
• T. Gur, I. Meydan, H. Seckin, M. Bekmezci, and F. Sen, "Green synthesis, characterization and bioactivity of biogenic zinc oxide nanoparticles," (in English), Environmental Research, vol. 204, 2022, doi: 10.1016/j.envres.2021.111897.
• E. Turunc, R. Binzet, I. Gumus, G. Binzet, and H. Arslan, "Green synthesis of silver and palladium nanoparticles using Lithodora hispidula (Sm.) Griseb. (Boraginaceae) and application to the electrocatalytic reduction of hydrogen peroxide," (in English), Materials Chemistry and Physics, vol. 202, pp. 310-319, 2017, doi: 10.1016/j.matchemphys.2017.09.032.
• M. Fayadoglu, E. Fayadoglu, S. Er, A. T. Koparal, and A. S. Koparal, "Determination of biological activities of nanoparticles containing silver and copper in water disinfection with/without ultrasound technique," (in English), Journal of Environmental Health Science and Engineering, vol. 21, no. 1, pp. 73-83, 2023, doi: 10.1007/s40201-022-00839-6.
• S. Pehlivanoglu, C. A. Acar, and S. Donmez, "Characterization of green synthesized flaxseed zinc oxide nanoparticles and their cytotoxic, apoptotic and antimigratory activities on aggressive human cancer cells," (in eng), Inorganic & Nano-Metal Chemistry, vol. 53, no. 9, pp. 1022-1031, 2023, doi: 10.1080/24701556.2021.1980034.
• M. Silambarasan, S. Saravanan, and T. Soga, "Raman and photoluminescence studies of Ag and Fe-doped ZnO nanoparticles," (in English), International Journal of ChemTech Research, vol. 7, no. 3, pp. 1644-1650, 2015. [Online]. Available: https://research.ebsco.com/linkprocessor/plink?id=c796b505-73d3-3a21-adfd-fd73e12ecbce.
• S. Martin, H. Takao, K. Tomoyuki, and H. Yoshikuni, "Determination of SiO2 Raman spectrum indicating the transformation from coesite to quartz in Gföhl migmatitic gneisses in the Moldanubian Zone, Czech Republic," vol. 103, ed, 2008, p. 105.
• K. P. Ashok, D. Sarita, and T. Sachin, "Synthesis and Characterization of Zinc Oxide Nanoparticles for Ethanol Detection," in Physics of Semiconductor Devices ISBN: 9783319030012, ed: Springer International Publishing, 2014.
• A. Sirelkhatim et al., "Review on Zinc Oxide Nanoparticles: Antibacterial Activity and Toxicity Mechanism," (in English), Nano-Micro Letters, vol. 7, no. 3, pp. 219-242, 2015, doi: 10.1007/s40820-015-0040-x.
• C. R. Mendes et al., "Antibacterial action and target mechanisms of zinc oxide nanoparticles against bacterial pathogens," (in English), Scientific Reports, vol. 12, no. 1, 2022, doi: 10.1038/s41598-022-06657-y.
• V. G. Sergey, E. B. Dmitriy, A. S. Dmitriy, B. R. Maxim, A. S. Anastasia, and B. L. Andrey, "A Mini Review of Antibacterial Properties of ZnO Nanoparticles," in Frontiers in Physics vol. 9, ed: Frontiers Media S.A., 2021.
• S. Hamed and M. Emara, "Antibacterial and Antivirulence Activities of Acetate, Zinc Oxide Nanoparticles, and Vitamin C Against <i>E. coli</i> O157:H7 and <i>P. aeruginosa</i>," (in English), Current Microbiology, vol. 80, no. 2, 2023, doi: 10.1007/s00284-022-03151-6.
• J. Bindia et al., "Pharmacological assessment of Co3O4, CuO, NiO and ZnO nanoparticles via antibacterial, anti-biofilm and anti-quorum sensing activities," in Water Science and Technology vol. 87, ed: IWA Publishing, 2023, pp. 2840-2851.