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Nguyen, N.P.U.; Dang, N.T.; Doan, L.; Nguyen, T.T.H. Synthesis of Silver Nanoparticles. Encyclopedia. Available online: (accessed on 14 June 2024).
Nguyen NPU, Dang NT, Doan L, Nguyen TTH. Synthesis of Silver Nanoparticles. Encyclopedia. Available at: Accessed June 14, 2024.
Nguyen, Ngoc Phuong Uyen, Ngoc Tung Dang, Linh Doan, Thi Thu Hoai Nguyen. "Synthesis of Silver Nanoparticles" Encyclopedia, (accessed June 14, 2024).
Nguyen, N.P.U., Dang, N.T., Doan, L., & Nguyen, T.T.H. (2023, September 13). Synthesis of Silver Nanoparticles. In Encyclopedia.
Nguyen, Ngoc Phuong Uyen, et al. "Synthesis of Silver Nanoparticles." Encyclopedia. Web. 13 September, 2023.
Synthesis of Silver Nanoparticles

Silver nanoparticles, also known as AgNPs, have been extensively researched due to their one-of-a-kind characteristics, including their optical, antibacterial, and electrical capabilities. In the era of the antibiotics crisis, with an increase in antimicrobial resistance (AMR) and a decrease in newly developed drugs, AgNPs are potential candidates because of their substantial antimicrobial activity, limited resistance development, and extensive synergistic effect when combined with other drugs.

AgNPs biological synthesis silver nanoparticles

1. Physical Methods

Laser ablation, irradiation, evaporation and condensation, and lithography are the most important physical processes for producing AgNPs [1]. Energy used in this approach can be light energy (laser ablation method), electrical energy (electrical arc-discharge method), thermal energy (physical vapor deposition) and mechanical energy (ball milling method). The benefits of these procedures are a fast synthesis reaction rate, high purity due to the lack of solvent contamination, and an absence of dangerous compounds (i.e., reducing agents/stabilizers). The physical synthesis of AgNPs employs an atmospheric pressure tube furnace to create NPs by evaporation–condensation [2][3][4][5][6][7]. However, this process has several disadvantages. For instance, a tube furnace requires a large amount of storage space, a substantial amount of energy for heating, and considerable time to achieve thermal stability [8][9]. Laser ablation of metallic bulk materials in solution, electrical irradiation of an Ag target in pure water with a 532 nm laser beam, or nanosphere lithography can create small NPs with a narrow size distribution in pure water without the use of any chemical additives, and the benefit of being a simple and low-cost nanofabrication approach [10][11][12][13][14][15][16]. Aside from the benefits highlighted above, some of the disadvantages of these techniques are poor potential yields and significant energy consumption [15][17][18][19]. Overall, physical methods generate AgNPs with a fine size distribution. They are suitable for the large-scale production of a singular product while producing AgNPs in ash form efficiently [20].

2. Chemical Methods

To date, the most common methods for producing AgNPs are still based on chemical synthesis. Under certain conditions, Ag+ from a silver salt precursor, through electron transfer, is reduced to elemental silver (AgNPs) [21]. The processes of nucleation and growth take place to generate final the AgNP product. In brief, the concentration of silver element in the solution swiftly exceeds the supersaturation critical level, resulting in “burst nucleation” and precipitation, which leads to nucleus formation. Apart from nucleation, increased silver addition induces nuclei growth and the formation of larger NPs. These chemical AgNP synthesis techniques can produce NPs with no aggregation, a high yield, despite the high production costs and hazardous consequences [22][23]. In chemical synthesis procedures, the production of NPs requires three reactant components: a silver salt precursor, reducing agents, and a stabilizing chemical.
The function of stabilizing agents is to prevent agglomeration after synthesis. These reagents are usually surfactants containing functionalities serving as protectors, especially polymeric compounds, to coat AgNPs and protect the NP surface, prevent other AgNPs from absorbing on or binding to the NP surfaces leading to agglomeration [24]. However, Amir et al. demonstrated that the higher molar ratios for the stabilizer result in lower AgNPs due to an excess of stabilizer, preventing it from forming a complex with Ag+ [25].

2.1. Chemical Reduction

Numerous techniques have been devised for the chemical synthesis of AgNPs, including the chemical reduction method, the polyol method, and the radiolytic process. Chemical reduction using inorganic and organic reducing agents is the finest and simplest method for producing AgNPs without aggregation, with a high yield and minimal preparation cost [26]. Several reducing agents, including ascorbate, elemental hydrogen, polyol sodium borohydride (NaBH4), sodium citrate, and the Tollens reagent, are applied to reduce silver ions (Ag+) in aqueous or nonaqueous solutions. Principally, two elements are required for NP growth in this method: a silver salt and a reducing agent [27]. The silver ions are derived from silver salt such as silver nitrate, silver citrate and silver acetate. A reducing agent first reduces the ions to atoms: Ag+ (aq) + e → Ag (s) [28], then atoms are nucleated and grow into particles. The concentration ratio of silver salt to reducing agent determines the availability of atoms, which governs the size and morphology of the NPs. Higher silver salt concentrations reportedly generate more NPs for 24 h reaction times but if the reaction time increases to 96 h, AgNP population will not be homogenous. In this instance, the preponderance of Ag atoms is encapsulated within sizable NPs [29].
In case of mild reducing agent, the remaining Ag+ ions continued generated to Ag0 and firmly attached to the surface of existing Ag particles, causing the product’s morphology to change, quasi-spherical to polygonal shape for instances [27]. In other words, a sluggish rate of reaction leads to particle agglomeration, whereas a stronger reducing agent produces smaller AgNPs [30].The appearance of black sediment can indicate that AgNPs undergo agglomeration [31]. Recently, scientists have evaluated the use of cationic exchange resins to separate free silver ions from suspensions of synthesized AgNPs, thereby reducing the Ag+ content of an unprocessed suspension of AgNPs while preserving their integrity [32].

2.2. Microemulsion Techniques

Microemulsion techniques refer to the synthesis of AgNPs using surfactant for dispersing two immiscible liquids, such as oil and water, water and superficial CO2, a mixture of oil, one or few surfactants, and water [30]. This method can create homogeneous AgNPs with controllable size [24][33].
Basis for the preparation of AgNPs in two-phase aqueous organic systems is the initial spatial separation of reactants (Ag precursor and reducing agent) in two immiscible phases [34]. “Ready-to-use” surfactants could be anionic, cationic, zwitterionic, and nonionic reagents including bis(2-ethyhexyl) sulfosuccinate, lauryl sodium sulphate, sodium dodecylbenzene sulfonate (SDS) (anionic), cetyltrimethylammonium bromide (CTAB) and polyvinylpyrrolidone (PVP) (cationic) and Triton X-100 (non-ionic) [35]. The selection of surfactants should be determined by the requirements of the experiment and the reaction conditions. Different surfactants, or microemulsion systems, used in the fabrication process will produce AgNPs with distinct diameters or morphologies. The rate of interactions between metal precursors and reducing agents is influenced by the interface between the two liquids as well as the intensity of interphase transport, which is mediated by ammonium salt. The formation of silver clusters at the interface is stabilized by the transfer of non-polar aqueous medium stabilizer molecules to the organic medium by the interphase transporter [24][30][36].
There are several influent parameters having ability to affect the shape or the size of AgNPs: the type of continuous phase, the amount of precursor dissolved within the nanodroplets, and the amount of water, referred to as the molar ratio of water to surfactant (W), etc. [37]. A large number of NPs with small diameters are created by the high exchange rate between micelles. In contrast, slow material exchange between micelles results in the formation of fewer nuclei and a larger ultimate particle size [38]. For instance, using borohydrate as a reducing agent and a biosurfactant extracted from Pseudomonas aeruginosa MKVIT3 resulted in the generation of cubic AgNPs having size of 17.89 and 8.74 nm [39]. Meanwhile, it was also reported that at 70 °C, a combination of silver acetate and the reducing agent oleylamine can yield highly monodisperse AgNPs, which have a size ranging from 10 to 20 nm and a storage stability of 6 months [30][39][40].

2.3. Photochemical Method

The photochemical process begins with metal precursors to be reduced from n+ valence (Mn+) to zero-valence (M0) via the photocatalytic action of a reducing agent. The M0 creates nucleation centers or nuclei, which proliferate and aggregate into metallic NPs [41][42]. Ultraviolet light, sunlight, and laser light are examples of light sources, with ultraviolet light being the most prevalent [43]. Photochemical synthesis permits the formation of NPs in a variety of media, including cells, emulsions, polymer films, surfactant micelles, and glassware [24]. Among the variables that can influence the synthesis of AgNPs are the light’s source, intensity, and wavelength, as well as the irradiation duration. For instance, increasing duration and intensity of irradiation have been shown to promote Ag+ reduction [41][42][43].
Photochemical routes, as shown in Figure 1, in nanotechnology take more advantages over other methods, because they do not use toxic or hazardous compounds or require expensive equipment and highly trained personnel. They can be conducted at room temperature and atmospheric pressure [41].
Figure 1. Photochemical formation of NPs [41]. Created with

2.4. Polymers and Polysaccharides

To create AgNPs, water was used as an eco-friendly solvent, and polysaccharides were used as capping/reducing agents. Starch (the capping agent) and D-glucose (the reducing agent) were used to synthesize starch-AgNPs in a mildly heated system. Due to the feeble binding between starch and AgNPs, this linking is subsequently reversible at higher temperatures, and AgNPs are separated [24][42][44]. One of the most common polymers to use as reducing agents in synthesizing AgNPs is polyvinyl alcohol (PVA), polyethylene glycol (PEG), polyacrylamide, or other biopolymers such as chitosan [45][46][47]. PEG-coated AgNPs were reported to be extremely stable in extremely salty solutions, whereas carboxyl-coated lipoic acid particles can be used for bioconjugation [48]. In the case of PEG, ascorbic acid, thiosulfate, and sodium citrate were commonly used as reducing agents combination to create a specific shape of AgNPs, typically spherical AgNPs [30]. Meanwhile, stabilizing agents usually include polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), bovine serum albumin (BSA), citrate, and cellulose [49].
Currently, several AgNP polymer hybridization strategies have been proposed to improve the antimicrobial activity of AgNPs. The most interesting and typical compound now is chitosan-AgNPs, since the chitosan’s amine groups can alter bacterial cell permeability, improving the antibacterial activity of AgNP at low cost [50][51]. Because of metal ion coordination by the amino groups on the biopolymer chain, the principal amino groups of chitosan interact with metal surfaces and serve as capping sites for NP stabilization [50][52]. Silver ion reduction can occur via oxidation of other functional groups on the chitosan structure (e.g., hydroxyl groups), according to the mechanism of synthesis. The affinity between chitosan and AgNPs may be influenced by the formation of chemical bonds between nitrogen atoms, which are electron-rich elements, and the lone pairs in the silver orbitals [50][51]. Recently, a novel polyquaternary phosphonium oligochitosans (PQPOCs) linked with AgNPs. PQPOCs–AgNPs exhibited greater antiviral activity against norovirus (Nov), hepatitis A virus (HAV), and coxsackievirus B4 (Coxb4) than PQPOCs alone. The combination of PQPOCs-AgNPs is supposed to enhance molecular interactions with viral glycoproteins and block viral penetration [53][54].

2.5. Electrochemical Synthetic Method

In this method, AgNPs are formed under conditions of electrochemical discharge/ plasma in aqueous solutions in four main steps or reactions. Reaction 1, metallic ions are formed. In reaction 2, electrons are generated in microplasmas to readily hydrate in aqueous medium and can subsequently reduce metallic ions in reaction 3, so called electrochemical reduction. In reaction 4, nucleation of NPs takes place where metal atoms are combined into nanoclusters, then it is followed by growth and agglomeration that leads to the formation of NPs [55].
Reaction 1: Mn+ + ne → M0
Reaction 2: e + nH2O → eaq
Reaction 3: Mn+ + neaq → M0
Reaction 4: M0 → M2 → … → Mx → … → Magglomerate

2.6. Microwave-Assisted Synthesis

Microwave-assisted synthesis was discovered for the first time in the early 1940s. The procedure entails swiftly heating the silver precursor with microwave irradiation, which may facilitate nuclei generation on-site. Thus, control of the nucleation and growth phases of AgNP synthesis is improved. Data showed that AgNPs produced by microwave-assisted synthesis have a narrow size distribution and a high degree of crystallization [30][56][57].
Several variables can impact the microwave-assisted synthesis of AgNPs. They include precursor concentration, stabilizer type and chirality of reducing agents. Water and alcohol are the optimal medium for microwave heating stabilizers due to their high dielectric losses. Polar molecules, such as water, attempt to align the electric field in a microwave. When dipolar molecules attempt to reorient themselves relative to an alternating electric field, they lose energy as heat, which may contribute to the reduction of Ag+ [24][43]. Further, microwave power input, irradiation time, dielectric constant and medium refractive index can also influence the outcome of the synthesis process.
In addition to silver, microwave energy can also be used to synthesize silver-doped lanthanum chromites. It is possible to produce AgNPs by combining microwave energy and thermal reduction, which can then be deposited on oxidized carbon paper electrodes. This procedure yields AgNPs with homogenous particle sizes that are evenly distributed across a carbon paper substrate [24][43][58]. Despite of various chemical methods to synthesize AgNPs, each method has its own advantages and disadvantages, as shown in Table 1.
Table 1. Advantages and disadvantages of different chemical methods.

3. Green Chemistry Approach for the Synthesis of AgNPs

Chemical remnants of the solvent are frequently found on the surface of the synthesized AgNPs such as ethylene glycol, sodium citrate, oleyl amine, liquid paraffin. As hazardous chemical carriers, these NPs are especially hazardous when used for drug delivery, antimicrobial action, or any other application requiring insertion of the NPs inside the human body [62]. The chemical synthesis of AgNPs on a large scale is not feasible in a world seeking to attain sustainable development objectives. Due to the danger of chemical methods, it is urgent to develop an alternative synthesis pathway that is economical and eco-friendly, biological or “green” method, as shown in Figure 2. Green synthesis can be classified as (a) the use of microorganisms such as fungi, yeasts (eukaryotes), bacteria, and actinomycetes (prokaryotes), (b) the use of vegetation and plant compounds, and (c) the utilisation of templates such as membranes, virus DNA, and diatoms [63].
Figure 2. Biological approaches in AgNP synthesis. Created with
For the ecological synthesis of AgNPs, a reducing biological agent is applied. In the majority of instances, different cell constituents serve as stabilizing and capping agents, eliminating the need for external capping and stabilizing agents [64][65].
The green synthesis mechanism can be described briefly as follows: Organic chemicals donate electrons for the reduction of Ag+ ions to Ag0. These organic chemicals can be proteins, enzymes & coenzymes, lipids, carbohydrates, alkaloids, flavonoids, phenols, terpenoids, and others. The active ingredient responsible for Ag+ ion reduction varies depending on the used organism or biological extract. For the nano-transformation of AgNPs, electrons are believed to be derived from dehydrogenation of acids and alcohols in hydrophytes, keto to enol conversions in mesophytes, or both mechanisms in xerophytes. Similar reduction processes can be performed by microbial cellular and extracellular oxidoreductase enzymes [65].
Mohan and colleagues described a simple method for synthesis of AgNPs using Acacia gum (0.5% w/v) in a mild environment [66]. In this method, the carboxylate group of the acacia gum glycoprotein was converted into COOAg through an ion-exchange process then acacia gum polysaccharide polymer chains will facilitate the stabilization of formed AgNPs [67].

3.1. Plants

Plant-based synthesis of AgNPs is widely used in comparison to microorganism-based techniques because it is more effective, less bio-compromising, and does not require active cell cultures. Various plant parts such as bark, callus, flower, fruit, leaves, peel, rhizome, stem, and seed can be extracted to synthesize AgNPs [43][48] and AgNPs’ shapes produced from plants are commonly spherical or oval [68]. They contain a high concentration of carbohydrates, enzymes, flavonoids, polyphenols, and proteins. In the cell- free metallic NPs synthesis, these phytochemicals are extracted and used directly as reducing and stabilizing agents, thus supplanting potentially hazardous compounds such as sodium borohydride (NaBH4). Due to the presence of an extensive variety of phytoconstituents in the extracts, the precise mechanism underlying this phenomenon remains unknown. Although organic acids, polyphenols and proteins are believed to be the primary reducing agents, it is anticipated that the numerous phytochemicals collaborate. In general, this method is considered as a cost-effective solution for mass production [69][70][71]. Some of the active phytochemicals that may be responsible for the reduction of Ag+ to AgNPs are terpenoids, polysaccharides, phenolics, alkaloids, flavones, amino acids, alcoholic compounds, enzymes, and. With non-pathogenic nature, and eco-friendly reaction conditions, highly economical single step protocol, synthesizing AgNPs using plant extracts in green synthesis have been identified to be speedier than that of microorganisms such as bacteria and fungi and as an ideal candidate [63][68]. Furthermore, AgNPs synthesized from plants seemed to have higher bioactivity than the chemically synthesized one. Sreelekha et al. (2021) synthesized AgNPs from M. frondosa leaf extract and sodium citrate, DPPH (1, 1-Diphenyl-2-picrylhydrazyl) assay was used to evaluate the antioxidant activity of two methods. Green NPs exhibited 91% scavenging activity at a concentration of 5 μg/mL, whereas chemically synthesized NPs exhibited only 79% activity at the same concentration. The antioxidant properties of flavonoids and phenolic compounds—present on the surface of green synthesized NPs—make them useful for the prevention and treatment of degenerative diseases [72]. Moreover, AgNPs from Brachychiton populneus showed a significant effect on cancer cell, specifically, on the U87 and HEK 293 cell lines. As the concentration increases, the proportion of viable cancer cells decreases [73].

3.2. Microorganisms


Bacteria are of great interest for NP synthesis, even though this process is fraught with difficulties, such as culture contamination, long procedures, and limited control over NP size. Due to their exceptional ability to reduce heavy metal ions, microbes are regarded as one of the most promising candidates for NP synthesis [68][74]. Bacterial ability to survive in an extremely silver-rich environment may contribute to AgNP accumulation [75][76]. Because of the ease of recovery of AgNPs, the extracellular method is preferable to the intracellular method [43]. Exopolysaccharide, peptides, reductase, cofactors, c-type cytochromes, and silver-resistant genes are examples of organic substances that can be used as reducing agents in bacteria. Bacteria which have been used to synthesize AgNPs includes Lactobacillus bulgaricus [77], Rhodococcus, Brevundimonas and Bacillus [78], Staphylococcus aureus [79], and Escherichia coli [80]. Several enzymes, including nitrate reductase and lactate dehydrogenase, have been implicated in the formation of AgNPs. Special amino acids such as arginine, cysteine, lysine, methionine may adhere to the surface of the particle and function as reducing agents [43].

Algae and Fungi

Algae grow quickly, are simple to harvest and can be scaled up easily. Algae are rich in pigments, peptides, proteins, and secondary metabolites [30][43]. With abundance of organic compounds, algae are an excellent candidate for AgNP biosynthesis [81]. These active organic matters can be used as reducing agents to create spheres [82], triangles [83], cubes [84], hexagons [85] AgNPs, etc.
The ability of fungi to decompose organic matter is well known using a variety of enzymes and molecules [86]. A large number of fungal proteins and enzymes, including protease, cellulase, chitinase, and glucosidase, can directly accelerate and increase AgNP synthesis. NADH and NADH-dependent nitrate reductase are involved in the reduction of Ag+ ions to Ag0 by fungi. Fungi can also synthesize AgNPs intra- or extracellularly with no toxic chemicals required. NPs produced by intracellular synthesis are smaller than those produced by extracellular synthesis. Intracellular synthesis is preferable for several reasons, including fewer purification steps needed. Filamentous fungi are of higher interest than other fungi because AgNPs they produce have favorable morphological properties, stability, and a broad range of applications [30]. Depending on the fungus used, parameters such as agitation, temperature, light, and culture and synthesis periods can be altered to achieve the desired NP characteristics [87].


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