Green Synthesis of Silver Nanoparticles Using  Pistacia eurycarpa  Leaf Extract: Optimization, Characterization, and  In Vitro  Antioxidant Activity Evaluation

Israa Issa Hassan; Najlaa Kadhim Issa; Omer Hasan Azeez

doi:10.22146/ijc.116833

Green Synthesis of Silver Nanoparticles Using Pistacia eurycarpa Leaf Extract: Optimization, Characterization, and In Vitro Antioxidant Activity Evaluation

https://doi.org/10.22146/ijc.116833

Israa Issa Hassan^(1*), Najlaa Kadhim Issa⁽²⁾, Omer Hasan Azeez⁽³⁾

(1) Basic Medical Science Unit, College of Nursing, University of Duhok, Zakho Street 38, 1006 AJ Duhok, Kurdistan Region 42001, Iraq
(2) Department of Chemistry, College of Science, University of Duhok, Zakho Street 38, 1006 AJ Duhok, Kurdistan Region 42001, Iraq
(3) Department of Obstetrics, College of Veterinary Medicine, University of Duhok, Zakho Street 38, 1006 AJ Duhok, Kurdistan Region 42001, Iraq
(*) Corresponding Author

Abstract

Green synthesis of silver nanoparticles (AgNPs) using plant extracts offers a sustainable alternative to conventional chemical methods; however, studies on Pistacia eurycarpa remain limited, particularly in terms of mechanistic insights and process optimization. In this study, AgNPs were synthesized using P. eurycarpa leaf extract as a reducing and stabilizing agent. Optimal conditions were achieved at 75 °C, pH 11, 45 min, and 10 mM silver nitrate. UV-vis spectroscopy confirmed nanoparticle formation with a surface plasmon resonance peak at 420 nm. FTIR spectroscopy confirmed phenolic and flavonoid functional groups involved in nanoparticle formation, while HPLC identified rutin (88.9 ppm) and gallic acid (80.2 ppm) as major compounds responsible for silver ion reduction through electron donation and nanoparticle stabilization via surface capping. SEM showed spherical, well-dispersed nanoparticles with sizes of 6.2–36.9 nm, while XRD confirmed a crystalline face-centered cubic structure with crystallite sizes of 3.7–7.4 nm. The nanoparticles exhibited enhanced antioxidant activity, including a 57% increase in total antioxidant capacity, a 37% increase in reducing power, 94.15% radical scavenging activity, and an 11% improvement in iron chelation. This enhancement is attributed to increased surface area, improved electron transfer, and synergistic effects between nanoparticles and bioactive compounds.

Keywords

Pistacia eurycarpa; silver nanoparticles; green synthesis; characterization; optimization

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References

[1] Eker, F., Akdaşçi, E., Duman, H., Bechelany, M., and Karav, S., 2025, Green synthesis of silver nanoparticles using plant extracts: A comprehensive review of physicochemical properties and multifunctional applications, Int. J. Mol. Sci., 26 (13), 6222.

[2] Gonçalves, A.C., Pinto, A.R., Cima, A., Olo-Fontinha, E., Martins, J.C.L., Garcia, J., Lemos, A., Saavedra, M.J., Pintado, M.M., and Alves, M.J., 2026, Plant-derived bioactive compounds: One health perspective, Appl. Sci., 16 (1), 327.

[3] Fatiqin, A., Alfanaar, R., Rahman, S., Febrianto, Y., Citrariana, S., Arsana, M.P., Suprayogi, T., and Kurniawan, Y.S., 2024, Catalytic reduction of 4-nitrophenol and methylene blue with silver nanoparticles decorated with Drymoglossum piloselloides extract, J. Multidiscip. Appl. Nat. Sci., 4 (2), 249–261.

[4] Kakodkar, S., Dhawal, P., Kadam, J., Khan, R., Shewale, P., and Chaukekar, T., 2025, Biosynthesis of silver nanoparticles from Callistephus chinensis flower waste: Evaluation of antibacterial, anticancer, and contaminated water remediation applications, J. Multidiscip. Appl. Nat. Sci., 5 (3), 763–782.

[5] Anugrahwati, M., Wahyuni, E.T., and Sudiono, S., 2026, Green synthesis of AgNPs from argentometric waste using Sandoricum koetjape pericarp extract: Structural characterization and photocatalytic activity, J. Multidiscip. Appl. Nat. Sci., 6 (1), 91–107.

[6] Wang, X., Wang, W., Ejaz, A., Mehmood, M., Ahmad, N., Nazir, I., Shahzad, Y., and Bai, T., 2025, Green synthesis of Colocasia esculenta based silver nanoparticles: Characterization and transdermal delivery for anti-inflammatory applications, Front. Pharmacol., 16, 1611507.

[7] Almatroudi, A., 2020, Silver nanoparticles: Synthesis, characterisation and biomedical applications, Open Life Sci., 15 (1), 819–839.

[8] Fatiqin, A., Amrulloh, H., Simanjuntak, W., Apriani, I., Amelia, R.A.H.T., Syarifah, S., Sunarti, R.N., and Raharjeng, A.R.P., 2021, Characteristics of nano-size MgO prepared using aqueous extract of different parts of Moringa oleifera plant as green synthesis agents, AIP Conf. Proc., 2331 (1), 040001.

[9] Fatiqin, A., Alfanaar, R., Rahman, S., Febrianto, Y., Citrariana, S., Arsana, M.P., Suprayogi, T., Kurniawan, Y.S., and Amrulloh, H., 2026, Antibacterial activities of silver nanoparticles prepared using extract of Drymoglossum piloselloides, J. Multidiscip. Appl. Nat. Sci., 6 (1), 455–465.

[10] Singhal, G., Bhavesh, R., Kasariya, K., Sharma, A.R., and Singh, R.P., 2011, Biosynthesis of silver nanoparticles using Ocimum sanctum (Tulsi) leaf extract and screening its antimicrobial activity, J. Nanopart. Res., 13 (7), 2981–2988.

[11] Gil-Korilis, A., and Ergui-Arbizu, J., 2024, A ride on the current state of silver nanoparticles in health: What is the next stop?, Bioactivities, 2 (2), 96–106.

[12] Abdel-Alim, M.E., Samaan, K., Guillaume, D., and Amla, H., 2023, Green synthesis of silver nanoparticles using Egyptian date palm (Phoenix dactylifera L.) seeds and their antibacterial activity assessment, Bioactivities, 1 (1), 1–8.

[13] Giri, A.K., Jena, B., Biswal, B., Pradhan, A.K., Arakha, M., Acharya, S., and Acharya, L., 2022, Green synthesis and characterization of silver nanoparticles using Eugenia roxburghii DC. extract and activity against biofilm-producing bacteria, Sci. Rep., 12 (1), 8383.

[14] Rauf, A., Patel, S., Uddin, G., Siddiqui, B.S., Ahmad, B., Muhammad, N., Mabkhot, Y.N., and Ben Hadda, T., 2017, Phytochemical, ethnomedicinal uses and pharmacological profile of genus Pistacia, Biomed. Pharmacother., 86, 393–404.

[15] Karcı, H., and Kafkas, S., 2024, The first complete chloroplast genome sequences of Pistacia eurycarpa Yalt, a horticultural important rootstock and industrial tree, Genet. Resour. Crop Evol., 71 (7), 3959–3972.

[16] Elez Garofulić, I., Kruk, V., Martić, A., Martić, I., Zorić, Z., Pedišić, S., Dragović, S., and Dragović-Uzelac, V., 2020, Evaluation of polyphenolic profile and antioxidant activity of Pistacia lentiscus L. leaves and fruit extract obtained by optimized microwave-assisted extraction, Foods, 9 (11), 1556.

[17] Salhi, A., Bellaouchi, R., El Barkany, S., Rokni, Y., Bouyanzer, A., Asehraou, A., Amhamdi, H., Zarrouk, A., and Hammouti, B., 2019, Total phenolic content and antimicrobial activities of Pistacia lentiscus leaves, Caspian J. Environ. Sci., 17 (3), 189–198.

[18] Singh, P., Kim, Y.J., Zhang, D., and Yang, D.C., 2016, Biological synthesis of nanoparticles from plants and microorganisms, Trends Biotechnol., 34 (7), 588–599.

[19] Mittal, A.K., Chisti, Y., and Banerjee, U.C., 2013, Synthesis of metallic nanoparticles using plant extracts, Biotechnol. Adv., 31 (2), 346–356.

[20] Whelan, L.C., Geary, M., and Healy, J., 2022, A novel, simple rapid reverse-phase HPLC-DAD analysis for the simultaneous determination of phenolic compounds and abscisic acid commonly found in foodstuffs and beverages, J. Chromatogr. Sci., 60 (7), 648–654.

[21] Mazarakioti, E.C., Zotos, A., Thomatou, A.A., Kontogeorgos, A., Patakas, A., and Ladavos, A., 2022, Inductively coupled plasma–mass spectrometry (ICP-MS), a useful tool in authenticity of agricultural products’ and foods’ origin, Foods, 11 (22), 3705.

[22] Gündeşli, M.A., 2020, Determination of sugar, total phenol contents-and antioxidant activity of various parts of “Uzun” pistachio cultivar (Pistacia vera L.), Int. J. Agric. Environ. Food Sci., 4 (1), 62–69.

[23] Chang, C.C., Yang, M.H., Wen, H.M., and Chern, J.C., 2002, Estimation of total flavonoid content in propolis by two complementary colorimetric methods, J. Food Drug Anal., 10 (3), 178–182.

[24] Abeysuriya, H.I., Bulugahapitiya, V.P., and Loku Pulukkuttige, J., 2020, Total vitamin C, ascorbic acid, dehydroascorbic acid, antioxidant properties, and iron content of underutilized and commonly consumed fruits in Sri Lanka, Int. J. Food Sci., 2020 (1), 4783029.

[25] Dawadi, S., Katuwal, S., Gupta, A., Lamichhane, U., Thapa, R., Jaisi, S., Lamichhane, G., Bhattarai, D.P., and Parajuli, N., 2021, Current research on silver nanoparticles: Synthesis, characterization, and applications, J. Nanomater., 2021 (1), 6687290.

[26] Özkan, S., Kabak, B., and Kendüzler, E., 2023, Optimization effect on green synthesis of silver nanoparticles (AgNPs) using Dimrit raisin extract and their antimicrobial activity, J. Inst. Sci. Technol., 13 (3), 1926–1936.

[27] Akhtar, W., Hamza, M.I., Qayyum, S., Khan, M.A., Mukhtar, N., Kamal, A., Sarwar, R., Nazish, M., Alrefae, A.F., Almutairi, M.H., and Naseem, M.T., 2025, Phyto-synthesis, characterization of silver nanoparticles from mint leaf extract and its evaluation in antimicrobial and pharmacological applications, BMC Plant Biol., 25 (1), 1072.

[28] Marinova, G., and Batchvarov, V., 2011, Evaluation of the methods for determination of the free radical scavenging activity by DPPH, Bulg. J. Agric. Sci., 17 (1), 11–24.

[29] Umamaheswari, M., and Chatterjee, T.K., 2008, In vitro antioxidant activities of the fractions of Coccinia grandis L., leaf extract, Afr. J. Tradit., Complementary Altern. Med., 5 (1), 61–73.

[30] Mathew, S., and Abraham, T.E., 2006, Studies on the antioxidant activities of cinnamon (Cinnamomum verum) bark extracts, through various in vitro models, Food Chem., 94 (4), 520–528.

[31] Gulcin, I., and Alwasel, S.H., 2022, Metal ions, metal chelators and metal chelating assay as antioxidant method, Processes, 10 (1), 132.

[32] de Carvalho, N.K.G., Duarte Leite, D.O., Colares, A.V., Souza, F.A., Calabrese, K.D., Torres Salazar, G.J., Nascimento, J.B., Pereira da Silva, M., Rodrigues, F.F., and Costa, J.G., 2025, Quantification of phenolic compounds by HPLC/DAD and evaluation of the antioxidant, antileishmanial, and cytotoxic activities of ethanolic extracts from the leaves and bark of Sarcomphalus joazeiro (Mart.), Plants, 14 (11), 1733.

[33] Van Acker, T., Theiner, S., Bolea-Fernandez, E., Vanhaecke, F., and Koellensperger, G., 2023, Inductively coupled plasma mass spectrometry, Nat. Rev. Methods Primers, 3 (1), 52.

[34] Chen, W., Yang, Y., Fu, K., Zhang, D., and Wang, Z., 2022, Progress in ICP-MS analysis of minerals and heavy metals in traditional medicine, Front. Pharmacol., 13, 891273.

[35] Garibo, D., Borbón-Nuñez, H.A., de León, J.N.D., García Mendoza, E., Estrada, I., Toledano-Magaña, Y., Tiznado, H., Ovalle-Marroquín, M., Soto-Ramos, A.G., Blanco, A., Rodríguez, J.A., Romo, O.A., Chávez-Almazán, L.A., and Susarrey-Arce, A., 2020, Green synthesis of silver nanoparticles using Lysiloma acapulcensis exhibit high-antimicrobial activity, Sci. Rep., 10 (1), 12805.

[36] Khan, M.Z.H., Tareq, F.K., Hossen, M.A., and Roki, M.N.A.M., 2018, Green synthesis and characterization of silver nanoparticles using Coriandrum sativum leaf extract, J. Eng. Sci. Technol., 13 (1), 158–166.

[37] Galatage, S.T., Hebalkar, A.S., Dhobale, S.V., Mali, O.R., Kumbhar, P.S., Nikade, S.V., and Killedar, S.G., 2021, “Silver Nanoparticles: Properties, Synthesis, Characterization, Applications and Future Trends” in Silver Micro-Nanoparticles - Properties, Synthesis, Characterization, and Applications, Eds. Kumar, S., Kumar, P., and Pathak, C.S., IntechOpen, London, UK.

[38] Szczyglewska, P., Feliczak-Guzik, A., and Nowak, I., 2023, Nanotechnology—General aspects: A chemical reduction approach to the synthesis of nanoparticles, Molecules, 28 (13), 4932.

[39] Iravani, S., Korbekandi, H., Mirmohammadi, S.V., and Zolfaghari, B., 2014, Synthesis of silver nanoparticles: Chemical, physical and biological methods, Res. Pharm. Sci., 9 (6), 385–406.

[40] Ahmed, S., Ahmad, M., Swami, B.L., and Ikram, S., 2016, A review on plants extract mediated synthesis of silver nanoparticles for antimicrobial applications: A green expertise, J. Adv. Res., 7 (1), 17–28.

[41] Velgosova, O., Mačák, L., Lisnichuk, M., and Varga, P., 2025, Influence of pH and temperature on the synthesis and stability of biologically synthesized AgNPs, Appl. Nano, 6 (4), 22.

[42] Alabrahim, O.A.A., Abdeldayem, A.M., and Azzazy, H.M.S., 2025, Green synthesis of metallic nanoparticles using Pistacia species: Improved stability and biological activities, Nanoscale Adv., 7, 5449–5478.

[43] Liaqat, N., Jahan, N., ur-Rahman, K., Anwar, T., and Qureshi, H., 2022, Green synthesized silver nanoparticles: Optimization, characterization, antimicrobial activity, and cytotoxicity study by hemolysis assay, Front. Chem., 10, 952006.

[44] Alim-Al-Razy, M., Asik Bayazid, G.M., Ur Rahman, R., Bosu, R., and Shamma, S.S., 2020, Silver nanoparticle synthesis, UV-vis spectroscopy to find particle size and measure resistance of colloidal solution, J. Phys.: Conf. Ser., 1706 (1), 012020.

[45] Flieger, J., Franus, W., Panek, R., Szymańska-Chargot, M., Flieger, W., Flieger, M., and Kołodziej, P., 2021, Green synthesis of silver nanoparticles using natural extracts with proven antioxidant activity, Molecules, 26 (16), 4986.

[46] Lina, J.M., Mostafa, M.S., Yeasmin, S., Tasnim, N., Dipto, A.R., Das, H., Amin, M.N., Khan, N.M.M.U., Habib, A., and Rahman, M., 2025, Green-synthesized silver nanoparticles for improved heat stress resilience and germination in potato seeds, RSC Adv., 15 (47), 39975–39987.

[47] Ali, M.H., Azad, M.A.K., Khan, K.A., Rahman, M.O., Chakma, U., and Kumer, A., 2023, Analysis of crystallographic structures and properties of silver nanoparticles synthesized using PKL extract and nanoscale characterization techniques, ACS Omega, 8 (31), 28133–28142.

[48] Asif, M., Yasmin, R., Asif, R., Ambreen, A., Mustafa, M., and Umbreen, S., 2022, Green synthesis of silver nanoparticles (AgNPs), structural characterization, and their antibacterial potential, Dose-Response, 20 (2), 15593258221088709.

[49] Amrulloh, H., Ichsan, C., Simanjuntak, W., Zuas, O., Kurniawan, Y.S., and Simonescu, C.M., 2026, Biological and photocatalytic activities of magnesium oxide nanoparticles prepared from seawater bittern by electrochemical method, J. Multidiscip. Appl. Nat. Sci., 6 (1), 481–495.

[50] Xu, L., Wang, Y.Y., Huang, J., Chen, C.Y., Wang, Z.X., and Xie, H., 2020, Silver nanoparticles: Synthesis, medical applications and biosafety, Theranostics, 10 (20), 8996–9031.

[51] Gul, A., Baig, M.N., Ahmed, D., Najam, Z., Aslam, T., and Ali, S., 2024, Green synthesis of silver nanoparticles from Spirulina platensis extract: Antibacterial and antioxidant potential, BioNanoScience, 14 (3), 2327–2336.

[52] Chaves, N., Santiago, A., and Alías, J.C., 2020, Quantification of the antioxidant activity of plant extracts: Analysis of sensitivity and hierarchization based on the method used, Antioxidants, 9 (1), 76.

[53] Balkrishna, A., Kumar, A., Arya, V., Rohela, A., Verma, R., Nepovimova, E., Krejcar, O., Kumar, D., Thakur, N., and Kuca, K., 2021, Phytoantioxidant functionalized nanoparticles: A green approach to combat nanoparticle-induced oxidative stress, Oxid. Med. Cell. Longevity, 2021 (1), 3155962.

DOI: https://doi.org/10.22146/ijc.116833