Biogenic Silver Nanoparticles (AgNPs) from Marphysa moribidii Extract: Optimization of Synthesis Parameters

Interest in biogenic silver nanoparticles (AgNPs) is steadily increasing due to the cost-effective, easy, and environmentally friendly way in which they are synthesized. Synthesis using polychaete (Marphysa moribidii) extract as a reducing agent is particularly new and has the potential of being applied in various industries. However, biogenic AgNPs require synthesis optimization to increase their stability, yield, and characteristics. To meet these requirements, several synthesis parameters (such as polychaete size (body width), silver nitrate (AgNO₃) concentration, pH of polychaete crude extract, and the temperature during pre-incubation) and storage conditions were optimized in this study. The optimized conditions for obtaining high yield and stable AgNPs were polychaetes with a body width of 6–8 mm, 1 mM AgNO₃ with polychaete crude extract of pH 9, preheated at 90°C for 15 min before incubation at 30°C (150 rpm) for 24 hours, and stored at 4°C for long-term stability. The formation of AgNPs was confirmed through observation of a color transition (from pinkish to yellowish-brown) and analysis of UV-Vis spectra (between 398 and 400 nm). Scanning electron microscopy and transmission electron microscopy revealed the formation of spherical AgNPs with an average size of approximately 40.19 nm. Further, the optimized AgNPs demonstrated high storage stability for up to 6 months without any agglomeration. It is believed that these parameters are eminently suitable for the production of stable biosynthesized AgNPs.

Biosynthesis; Marphysa moribidii; Optimization; Polychaetes; Silver nanoparticles

        Research into nanotechnology has garnered significant interest worldwide over the last few decades due to its superior physical, chemical, and biological properties compared to its bulk form (Usman et al., 2018; Khalil et al., 2019). One of the most promising nanoparticles in the scientific world is silver nanoparticles (AgNPs). 

These are 10 to 100 nm in size and have superior physicochemical and antimicrobial properties, mainly due to their large surface area per volume ratio (Jeevanandam et al., 2018). Accordingly, AgNPs have been incorporated in various fields, including diagnosis, therapeutics, electrical, antimicrobial action, and catalysis (Zhang et al., 2016; Lee and Jun, 2019; Nakamura et al., 2019).

Conventionally, AgNPs have been synthesized by utilizing physical and chemical methods, which are costly and produce substances that are harmful to organisms and the environment (Zhang et al., 2016). However, numerous chemical and physical methods have been substituted by biological approaches in the last few years (Nakamura et al., 2019) that are environmentally sustainable and economical, as no toxic or expensive chemicals are utilized in the synthesis (Lee and Jun, 2019). In biological synthesis, two main components are required to initiate synthesis: a silver precursor such as silver nitrate (AgNO₃) and a biomolecule cocktail extracted from living organisms that concurrently act as reducing and stabilizing agents (Siddiqi et al., 2018). These biomolecules react with silver ion (Ag⁺) and are reduced to AgNPs of various forms and sizes and act as capping agents to stabilize AgNPs (Zhang et al., 2016).

Microorganisms and plants are commonly exploited as reducing agents in this method since they are widely available in nature (Lee and Jun, 2019; Nakamura et al., 2019). Synthesizing AgNPs from invertebrate marine sources (especially polychaetes) is a relatively recent process. The polychaete is commonly referred to as a segmented worm (or marine worm) and belongs to the class of Polychaeta (Phylum Annelida). Polychaetes are generally used as pollution indicators, fish broodstock, and fish bait (Gaete et al., 2017; Cole et al., 2018; Masimen et al., 2020). Previous studies have indicated that some marine polychaetes can synthesize AgNPs (Singh et al., 2014; Hussain et al., 2018). However, synthesis was initiated utilizing species other than Marphysa moribidii (locally known as ‘ruat bakau’). This is from the Eunicidae family and is harvested locally to be used as bait worm for recreational or artisanal fisheries (Idris et al., 2014). Using M. moribidii as a reducing agent in AgNP synthesis can increase the polychaete’s commercial value, especially in Malaysia.

    This research has demonstrated the successful optimization of biogenic AgNPs synthesized from marine polychaete (M. moribidii). The protocol was optimized, offering rapid production of AgNPs with increased stability and characteristics. The present investigation concluded that the green synthesis of AgNPs utilizing marine invertebrates (M. moribidii) as reducing and stabilizing agents offered many advantages. These included being economical, straightforward, and eco-friendly, which can scale up economic viability. Future studies could consider exploring possible applications of biosynthesized AgNPs, especially in healthcare. The findings of this research can be useful as a reference for biogenic AgNPs and green nanotechnology research, with the aim of discovering more marine polychaetes with the potential to be utilized as reducing and stabilizing agents in AgNP synthesis.

    We would like to thank the Ministry of Higher Education, Malaysia, and the Universiti Malaysia Terengganu for their funding (FRGS/1/2016/WAB09/UMT/02/2).

Ahmad, B., Ali, J., Bashir, S., 2013. Optimization and Effects of Different Reaction Conditions for the Bioinspired Synthesis of Silver Nanoparticles using Hippophae rhamnoides Linn. Leaves Aqueous Extract. World Applied Sciences Journal, Volume 22(6), pp. 836–843

Amini, N., Amin, G., Azar, Z.J., 2017. Green Synthesis of Silver Nanoparticles using Avena sativa Leaves Extract. Nanomedicine Research Journal, Volume 2(1), pp. 57–63

Ashraf, J.M., Ansari, M.A., Khan, H.M., Alzohairy, M.A., Choi, I., 2016. Green Synthesis of Silver Nanoparticles and Characterization of their Inhibitory Effects on AGEs Formation using Biophysical Techniques. Scientific Reports, Volume 6(20414), pp. 1–10

Ayad, Z.M., Ibrahim, O.M.S., Omar, L.W., 2019. Biosynthesis and Characterization of Silver Nanoparticles by Silybum marianum (silymarin) Fruit Extract. Advances in Animal and Veterinary Sciences, Volume 7(2), pp. 122–130

Bhatnagar, S., Kobori, T., Ganesh, D., Ogawa, K., Aoyagi, H., 2019. Biosynthesis of Silver Nanoparticles Mediated by Extracellular Pigment from Talaromyces purpurogenus and their Biomedical Applications. Nanomaterials, Volume 9(7), pp. 1–20

Chowdhury, S., Yusof, F., Faruck, M.O., Sulaiman, N., 2016. Process Optimization of Silver Nanoparticle Synthesis using Response Surface Methodology. Procedia Engineering, Volume 148, pp. 992–999

Cole, V.J., Chick, R.C., Hutchings, P.A., 2018. A Review of Global Fisheries for Polychaete Worms as a Resource for Recreational Fishers: Diversity, Sustainability and Research Needs. Reviews in Fish Biology and Fisheries, Volume 28, pp. 543–565

El-Seedi, H.R., El-Shabasy, R.M., Khalifa, S.A.M., Saeed, A., Shah, A., Shah, R., Iftikhar, F.J., Abdel-Daim, M.M., Omri, A., Hajrahand, N.H., Sabir, J.S.M., Zou, X., Halabi, M.F., Sarhan, W., Guo, W., 2019. Metal Nanoparticles Fabricated by Green Chemistry using Natural Extracts: Biosynthesis, Mechanisms, and Applications. RSC Advances, Volume 9(42), pp. 24539–24559

Gaete, H., Álvarez, M., Lobos, G., Soto, E., Jara-Gutiérrez, C., 2017. Assessment of Oxidative Stress and Bioaccumulation of the Metals Cu, Fe, Zn, Pb, Cd in the Polychaete Perinereis gualpensis from Estuaries of Central Chile. Ecotoxicology and Environmental Safety, Volume 145, pp. 653–658

Górska, B., Gromisz, S., W?odarska?Kowalczuk, M., 2019. Size Assessment in Polychaete Worms—Application of Morphometric Correlations for Common North Atlantic Taxa. Limnology and Oceanography: Methods, Volume 17(4), pp. 254–265

Hussain, N.S., Harun, N.A., Radzi, M.N.F., Idris, I., Wan Ismail, W.I., 2018. Biosynthesis of Silver Nanoparticles from Marine Polychaete Diopatra claparedii Grube, 1878. Jurnal Teknologi, Volume 80(6), pp. 181–187

Idris, I., Hutchings, P., Arshad, A., 2014. Description of a New Species of Marphysa Quatrefages, 1865 (Polychaeta: Eunicidae) from the West Coast of Peninsular Malaysia and Comparisons with Species from Marphysa Group A from the Indo-West Pacific and Indian Ocean. Memoirs of Museum Victoria, Volume 71, pp. 109–121

Izak-Nau, E., Huk, A., Reidy, B., Uggerud, H., Vadset, M., Eiden, S., Voetz, M., Himly, M., Duschl, A., Dusinska, M., Lynch, I., 2015. Impact of Storage Conditions and Storage Time on Silver Nanoparticles’ Physicochemical Properties and Implications for their Biological Effects. RSC Advances, Volume 5(102), pp. 84172–84185

Jain, S., Mehata, M.S., 2017. Medicinal Plant Leaf Extract and Pure Flavonoid Mediated Green Synthesis of Silver Nanoparticles and their Enhanced Antibacterial Property. Scientific Reports, Volume 7(1), p. 1–13

Jeevanandam, J., Barhoum, A., Chan, Y.S., Dufresne, A., Danquah, M.K., 2018. Review on Nanoparticles and Nanostructured Materials: History, Sources, Toxicity and Regulations. Beilstein Journal of Nanotechnology, Volume 9(1), pp. 1050–1074

Karekalammanavar, G., David, M., 2018. Optimization of Green Synthesis of Silver Nanoparticles from Leaf Extracts of Tabernaemontana heyneana and Evaluation of their Catalytic Activity on Reduction of Methylene Blue. Haya: The Saudi Journal of Life Sciences, Volume 3(3), pp. 248–254

Khalil, M., Rahmaningsih, G., Gunlazuardi, J., Umar, A., 2019. The Influence of Plasmonic Au Nanoparticle Integration on the Optical Bandgap of Anatase TiO₂ Nanoparticles. International Journal of Technology, Volume 10(4), pp. 808–817

Lee, S., Jun, B.H., 2019. Silver Nanoparticles: Synthesis and Application for Nanomedicine. International Journal of Molecular Sciences, Volume 20(4), pp. 865–885

Lee, S.W., Chang, S.H., Lai, Y.S., Lin, C.C., Tsai, C.M., Lee, Y.C., Chen, J.C., Huang, C.L., 2014. Effect of Temperature on the Growth of Silver Nanoparticles using Plasmon-Mediated Method under the Irradiation of Green LEDs. Journal of Material, Volume 7(12), pp. 7781–7798

Masimen, M.A.A., Harun, N.A., Misbah, S., Idris, I., Wan Ismail, W.I., 2020. Marine Resources: Potential of Polychaete Application in Combating COVID-19 Infection. Journal of Sustainability Science and Management, Volume 15(7), pp. 1–7

Mishra, A., Kaushik, N.K., Sardar, M., Sahal, D., 2013. Evaluation of Antiplasmodial Activity of Green Synthesized Silver Nanoparticles. Colloids and Surfaces B: Biointerfaces, Volume 111, pp. 713–718

Muñoz, R.V., Jimenez, M.J.A, Lopez, F.D., Ribot, J.L.L., 2019. Protocol Optimization for a Fast, Simple and Economical Chemical Reduction Synthesis of Antimicrobial Silver Nanoparticles in Non-specialized Facilities. BMC Research Notes, Volume 12(1), pp. 773–779

Nair, S.S., Rizvi, S.T., Anthappan, P.D., 2018. Optimization of Rapid Green Synthesis of AgNPs using Citrus Sinensis Peel Extract for Antibacterial Activity. International Journal of Advanced Scientific Research and Management, Volume 3(8), pp. 274–281

Nakamura, S., Sato, M., Sato, Y., Ando, N., Takayama, T., Fujita, M., Ishihara, M., 2019. Synthesis and Application of Silver Nanoparticles (AgNPs) for the Prevention of Infection in Healthcare Workers. International Journal of Molecular Sciences, Volume 20(15), pp. 3620–3638

Ndikau, M., Noah, N.M., Andala, D.M., Masika, E., 2017. Green Synthesis and Characterization of Silver Nanoparticles using Citrullus lanatus Fruit Rind Extract. International Journal of Analytical Chemistry, Volume 2017, pp. 1–9

Occhioni, G.E., Brasil, A.C.S., Araújo, A.F.B., 2010. Morphometric study of Phragmatopoma caudata (Polychaeta: Sabellida: Sabellariidae). Zoologia (Curitiba), Volume 26(4), pp. 739–746

Pei, A.U.E., Huai, P.C., Masimen, M.A.A., Wan Ismail, W.I., Idris, I., Harun, N. A., 2020. Biosynthesis of Gold Nanoparticles (AuNPs) by Marine Baitworm Marphysa moribidii Idris, Hutchings and Arshad 2014 (Annelida: Polychaeta) and its Antibacterial Activity. Advances in Natural Sciences: Nanoscience and Nanotechnology, Volume 11(1), pp. 1–10

Raja, S., Ramesh, V., Thivaharan, V., 2017. Green Biosynthesis of Silver Nanoparticles using Calliandra haematocephala Leaf Extract, their Antibacterial Activity and Hydrogen Peroxide Sensing Capability. Arabian Journal of Chemistry, Volume 10(2), pp. 253–261

Raman, P.R., Parthiban, S., Srinithya, B., Kumar, V.V., Anthony, S.P., Sivasubramanian, A., Muthuraman, M.S., 2015. Biogenic silver Nanoparticles Synthesis using the Extract of The Medicinal Plant Clerodendron serratum and Its In-Vitro Antiproliferative Activity. Materials Letters, Volume 160, pp. 400–403

Rosman, N.S.R., Harun, N.A., Idris, I., Wan Ismail, W.I., 2020. Eco-friendly Silver Nanoparticles (AgNPs) Fabricated by Green Synthesis using the Crude Extract of Marine Polychaete, Marphysa moribidii: Biosynthesis, Characterization, and Antibacterial Applications. Heliyon, Volume 6(11), pp. 1–9

Ruttkay-nedecky, B., Skalickova, S., Kepinska, M., Cihalova, K., Docekalova, M., Stankova, M., Uhlirova, D., Fernandez, C., Sochor, J., Milnerowicz, H., Beklova, M., Kizek, R., 2019. Development of New Silver Nanoparticles Suitable for Materials with Antimicrobial Properties. Journal of Nanoscience and Nanotechnology, Volume 19(5), pp. 2762–2769

Saxena, J., Sharma, P.K., Sharma, M.M., Singh, A., 2016. Process Optimization for Green Synthesis of Silver Nanoparticles by Sclerotinia sclerotiorum MTCC 8785 and Evaluation of Its Antibacterial Properties. SpringerPlus, Volume 5(1), pp. 1–10

Shahzad, A., Saeed, H., Iqtedar, M., Hussain, S.Z., Kaleem, A., Abdullah, R., Sharif, S., Naz, S., Saleem, F., Aihetasham, A., Chaudhary, A., 2019. Size-Controlled Production of Silver Nanoparticles by Aspergillus fumigatus BTCB10: Likely Antibacterial and Cytotoxic Effects. Journal of Nanomaterials, Volume 2019, pp. 1–14

Siddiqi, K. S., Husen, A., Rao, R. A. K., 2018. A Review on Biosynthesis of Silver Nanoparticles and their Biocidal Properties. Journal of Nanobiotechnology, Volume 16(1), pp. 14–42

Singh, R., Sahu, S.K., Thangaraj, M., 2014. Biosynthesis of Silver Nanoparticles by Marine Invertebrate (Polychaete) and Assessment of Its Efficacy against Human Pathogens. Journal of Nanoparticles, Volume 2014, pp. 1–7

Usman, A., Kusrini, E., Widiantoro, A.B., Hardiya, E., Abdullah, N.A., Yulizar, Y., 2018. Fabrication of Chitosan Nanoparticles Containing Samarium Ion Potentially Applicable for Fluorescence Detection and Energy Transfer. International Journal of Technology, Volume 9(6), pp. 1112–1120

Vinayagam, R., Varadavenkatesan, T., Selvaraj, R., 2018. Green Synthesis, Structural Characterization, and Catalytic Activity of Silver Nanoparticles Stabilized with Bridelia retusa Leaf Extract. Green Processing and Synthesis, Volume 7(1), pp. 30–37

Velgosova, O., ?ižmárová, E., Málek, J., Kavuli?ova, J., 2017. Effect of Storage Conditions on Long-term Stability of Ag Nanoparticles Formed Via Green Synthesis. International Journal of Minerals, Metallurgy, and Materials, Volume 24(10), pp. 1177–1182

Verma, A., Mehata, M.S., 2016. Controllable Synthesis of Silver Nanoparticles using Neem Leaves and their Antimicrobial Activity. Journal of Radiation Research and Applied Sciences, Volume 9(1), pp. 109–115

Zhang, X.F., Liu, Z.G., Shen, W., Gurunathan, S., 2016. Silver Nanoparticles: Synthesis, Characterization, Properties, Applications, and Therapeutic Approaches. International Journal of Molecular Sciences, Volume 17(9), p. 1534