Biosynthesis of metallic nanoparticles using plant extracts and evaluation of their antibacterial properties

Document Type : Review Paper


1 Department of Chemistry, University of Zanjan, Zanjan, Iran

2 Department of Chemistry, University of Zanjan, Zanjan, Iran; Research Institute of Modern Biological Techniques, University of Zanjan, Zanjan, Iran


The increasing attention being paid to metallic nano particles (MNPs) is due to their intensive applications in different areas of science such as medicine, chemistry, agriculture, and biotechnology. The main methods for nanoparticle production are chemical and physical approaches that are often costly and potentially harmful to the environment. Since the eco-friendly synthesis of NPs with different chemical compositions, sizes, shapes and controlled dispersity is an important aspect of nano biotechnology and green nanotechnology, biosynthesis of nanoparticles has been proposed as a cost-effective and environmental-friendly alternative to chemical and physical methods. Plants contain abundant natural compounds such as alkaloids, flavonoids, saponins, steroids, tannins and other nutritional compounds. These natural products are derived from various parts of the plant such as leaves, stems, roots, shoots, flowers, barks, fruits and seeds. Since the plant extract contains various secondary metabolites, it acts as the reducing and stabilizing agent for the bioreduction reaction to synthesize the novel metallic nanoparticles. This approach has been actively pursued in recent years as an alternative, efficient, inexpensive, and environmentally safe method for producing nanoparticles with specified properties. The present review focuses on the synthesis of MNPs with particular emphasis on biological synthesis using plant extracts and most commonly proposed mechanisms regarding the antibacterial properties of nanoparticles.



Nanotechnology is “the design, characterization, production and application of structures, devices and systems by controlling shape and size at nanometer scale” [1]. Normally, in nanotechnology, the curiosity lies on the matter with at least one dimension measuring between 1 and 100 nm. Within this range, materials may have properties considerably different from those expected when they have larger dimensions. Nanoscience is a new interdisciplinary subject that depends on the fundamental properties of nano size objects [2, 3]. A nanometer (nm) is a billionth of a meter, 10-9. To demonstrate this, the size of a human hair is 80,000 nm wide and a virus is around 100 nm in diameter. About ten atoms in a line make up one nanometer because one atom is 0.1 nm in diameter.

Nanomaterials can show atom-like behaviors which result from higher surface energy due to their large surface area, while a bulk material has constant physical properties regardless of its size, at the nanoscale, this is often not the case [4]. Several well-characterized bulk materials have been found to possess most interesting properties when studied in the nanoscale. There are many reasons for this including the fact that nanoparticles possess a very high aspect ratio. Due to the specific characteristics such as size, distribution and morphology, nanoparticles exhibit completely new or improved property, which impact all aspects of human life. Nanotechnology has opened a wide opportunity in the area of materials science, and the incorporation of other branches, such as photochemistry and electrochemistry to better understand its properties [5].

The metallic nanoparticles have been studied expansively because of their unique physicochemical characteristics including catalytic activity, optical properties, electronic properties, antibacterial properties and magnetic properties [6-11]. Noble metallic NPs, such as Ag, Au, Pd, have enticed tremendous interest in the scientific community [12, 13]. These materials have given rise to a busy area of research, due to the wide variety of applications in fields such as biomedicine, catalysis, preparation of nanocomposites with tunable electrical conductivity, thermal conductivity, tensile strength, superior rigidity, hardness and erosion resistance. So, they can be used for manufacturing of satellite components, aircraft spares, industry parts and electronic segments such as microchips processors [14-17].

The metallic nanoparticles are most promising; they have good antibacterial properties due to their large surface-area-to-volume ratio, which is of much interest to researchers due to the growing microbial resistance against metal ions, antibiotics, and the development of resistant strains. Silver nanoparticles show effective antimicrobial property compared to other nanoparticles due to their extremely large surface area, which provides better contact with microorganisms. In this way, nanoparticles are attached to the cell membrane and penetrate into the bacteria. The bacterial membrane contains sulfur-containing proteins, and the silver nanoparticles interact with these proteins in the cell as well as with the phosphorus-containing compounds like DNA. The nanoparticles preferably attack the respiratory chain and cell division finally leading to cell death. The nanoparticles release silver ions in the bacterial cells enhancing their bactericidal activity [18].

The highlight of this review is related to the green synthesis using plant extract and characterization of Cu, Zn, Ag NPs techniques and their antibacterial properties.

Techniques for the synthesis of nanoparticles

Generally, the methods for the synthesis of nanoparticles are usually classified into two categories: the physical and chemical techniques [19]. Physical synthetic methods such as inert gas condensation, severe plastic deformation, high-energy ball milling and ultrasonic shot peeling can be used to synthesize metallic nanoparticles [20]. Several methods including the commonly grinding process and pyrolysis can be used for the physical synthesis of metallic nanoparticles. The grinding process is the most representative example of the physical methods, where highly efficient mills are used to separate the particles of nanometric sized. In grinding process, macroscale or microscale particles are ground by a size-reducing mechanism; e.g., an ordinary or a planetary ball mill. The resulting particles are subsequently air classified to recover oxidized nanoparticles. The factors that critically affect the properties of the resultant nanoparticles include the material and time of milling and the atmospheric medium. In pyrolysis, an organic precursor (either a liquid or a gas) is forced through an orifice at high pressure and is burned. The resulting ash is air classified to recover oxidized nanoparticles.

However, these techniques have not been a reliable system to obtain metallic nanoparticles because, generally, the obtained particles are larger than 100 nm, which could not be considered as nanometric size. Another disadvantage of physical methods is expensive and cumbersome for the large-scale production of nanoparticles [21]. A further drawback of “physical” approaches is enormous consumption of energy to maintain the high pressure and temperature used in the synthesis procedures.

Chemical methods involve the reduction of chemicals [22], electrochemical procedures [23], and reduction of phytochemicals [24], microemulsion, chemical coprecipitation, chemical vapor condensation, pulse electrodeposition [20]. A typical procedure involves growing nanoparticles in a liquid medium containing various reactants, in particular reducing agents, such as sodium borohydride or potassium bitartrate [25] or methoxy polyethylene glycol [26] or hydrazine [21]. To prevent the agglomeration of metallic nanoparticles, a stabilizing agent such as sodium dodecyl benzyl sulfate [21] or polyvinyl pyrrolidone [25] is also added to the reaction mixture. Most chemical methods used for the synthesis of nanoparticles are too expensive and also involved with the use of toxic and hazardous chemicals that are responsible for various biological risks.

Various chemical and physical methods have been reported in order to synthesis nanoparticles, but some of the chemical procedures involved in the synthesis of these nanoparticles use toxic solvents which could potentially generate unsafe and hazardous byproducts and often involve high energy consumption [27-29].

Nanoparticle synthesis methods can also be classified as “bottom-up” and “top-down” by the direction of the nanoparticle formation. The “bottom-up” reaction begins from atomic level through forming molecules; however, in the opposite technique described as “top-down” the scale of the resultant nanoparticles is larger, so that a mechanical process or the addition of acids is required to reduce the particle size. Usually, the “top-down” technique requires the use of complex and complicated instrumentation (Fig. 1) [30].

The development of green processes for the synthesis of nanoparticles has been evolving into an important branch of nanotechnology as green nanotechnology deals with the safe and eco-friendly methods for nanomaterials fabrication which is considered as an alternative for the conventional physical and chemical methods. Green nanotechnology is gaining importance due to the elimination of harmful reagents and provides the effective synthesis of expected products in an economical manner [11, 31-39]. Noble metallic nanoparticles (MNPs) such as silver, gold and platinum are widely applied in medicinal applications. There is a growing need to develop an environmentally friendly process for the synthesis of nanoparticles that does not employ toxic chemicals [40-42]. Thus, synthesizing nanoparticles by biological means, which has the advantages of nontoxicity, reproducibility in production, easy scaling-up, and well-defined morphology, has become a new trend in nanoparticle production. In particular, microorganisms and plants have been demonstrated as new resources with considerable potential for synthesizing nanoparticles. In the case of biological methods, nanoparticle synthesis using plant extracts is the most adopted method, because it is eco-friendly, the green production of nanoparticles, it can act as a source of several metabolites, it is much safer to handle, and easily available [37, 38, 43]. While fungi and bacteria require a comparatively longer incubation time in the growth media for reducing a metal ion, water-soluble phytochemicals do the same in a jiffy. Therefore, compared with bacteria or fungi, plant cells are among the suitable candidates for the synthesis of metallic nanoparticles. Plant-based synthesis of nanoparticles works at low temperatures and requires only modest and environmentally safe components [44]. In addition, the synthesis of nanoparticles using plants offers other advantages, such as the utilization of safer solvents, decreased use of dangerous reagents, milder response conditions, feasibility, and their adaptability in use for medicinal, surgical, and pharmaceutical applications [45]. Furthermore, physical requirements for their synthesis, including pressure, energy, temperature, and constituent materials, are trivial.

In the biosynthesis of metallic nanoparticles using plant extract, three important parameters are (1) metal salt, (2) a reducing agent, and (3) a stabilizing or capping agent for controlling the size of nanoparticles and preventing their aggregation [46]. Many biomolecules in plants such as proteins/enzymes, amino acids, carbohydrates, alkaloids, terpenoids, tannins, saponins, phenolic compounds, reducing sugar and vitamins [47] could be involved in bioreduction, formation and stabilization of metal nanoparticles. The reduction potential of ions and reducing the capacity of plants which depend on the presence of polyphenols, enzymes, and other chelating agents present in plants have critical effects on the amounts of nanoparticle production (Fig. 2).

Antibacterial activity of metallic nanoparticles

Metal-based nanoparticles represent an alternative for biomedical treatments, mainly in the fabrication of biomedical devices with antimicrobial coatings. A high antimicrobial activity of nanoparticles depends on the particle size that allows greater surface contact and a direct interaction with the membranes of pathogenic microorganisms. The antibacterial activities of metallic nanoparticles were found to be related to their shapes and sizes. Nanoparticles smaller than 10 nm interact with bacteria and produce electronic effects, which improve the reactivity of nanoparticles. Thus, it is proven that the bactericidal effect of nanoparticles is size dependent [48].

The cell membranes of the microorganisms interact with the medium, so metal NPs will have some interactions to release metal ions that interfere with the processes of the DNA replication, cell membrane formation, cell division, and so forth, of certain microorganisms such as bacteria, which results in an antimicrobial effect [48, 49].

Several studies propose that nanoparticles attach to the surface of cell membrane disturbing the permeability and respiration function of the cell [18, 24]. The damage to the cell may be caused by the interaction of nanoparticles with sulfur or phosphorus-containing biomolecules in the cell such as DNA. Therefore, sulfur-containing proteins in the membrane or inside cells and phosphorus-containing elements like DNA are likely to be preferential sites for binding Ag and Zn [50-52] or CuNPs. Other studies suggest that when bacteria are treated with silver ions, DNA tends to lose its ability to replicate. Lokini et al. [53] showed that AgNPs could destabilize the outer membrane and rupture the plasma membrane, thereby depleting intracellular ATP. The excellent antibacterial activity of silver nanoparticles is mainly attributed to their high surface area-to-volume ratio that enables greater presence of atoms on the surface and, in turn, greater contact with the environment. The smaller nanoparticles have more antibacterial activity that provide more surface exposure to the bacterial membrane [54]. In addition , these nanosized particles penetrate through cell membrane easier, interacting with intracellular materials and finally resulting in cell destruction in the process of multiplication. Luo et al. [55] documented that the nanoparticles induce oxidative stress to bacteria and induce ROS production. For example, the antibacterial activity could be explained based on reactive oxygen species (ROS) such as H2O2, hydroxyl radicals, singlet oxygen, and Zn2+ ions released on the surface of ZnO which cause severe damage to bacteria [56-58]. The generation of hydrogen peroxide (H2O2) from the surface of ZnO is considered as an effective mean for the inhibition of bacterial growth according to some studies [59]. It has been reported that both UV and visible light can activate ZnO and consequently, electron-hole pairs (e-/h+) can be created. The generation of H2O2 is explained as follows: the holes split the H2O molecule from the suspension of ZnO into OH- and H+. Furthermore, dissolved oxygen molecules are converted to superoxide radical anions (O2•-) which react with hydrogen ion (H+) to produce HO2 radicals. The collision of these hydroxyl radicals with electrons will produce hydrogen peroxide anions HO2-, which react with hydrogen to generate H2O2 molecules. Therefore, the H2O2 molecules generated can penetrate into the cell membrane and kill the bacteria [60, 61]. The e-/h+ pair recombination minimizes the chances of ROS generation. Lattice defects play an important role in the inhibiting e-/h+ pair recombination process [62]. These defects may act as trapping centers and inhibit photo induced e-/h+ pair recombination [63] resulting in higher antibacterial activity. Further, under normal circumstances, cells are able to defend themselves against ROS damage with antioxidant enzymes. However, when the nanoparticles are inside the cell, the nanoparticles could restrain antioxidative enzymes to inhibit the capability of removing ROS. Also reported that the nanoparticles break the balance of oxidant/antioxidant and generate the accumulation of ROS in bacteria. However, exact mechanism of AgNPs on different bacterial cells needs a further study. Priester et al. [64] suggested that the action mechanism of the copper NPs occurs through the interaction of enzymes and -SH groups causing damage in the DNA and therefore oxidative stress generation [65-67]. The mechanism of penetration of nanoparticles to the bacteria is not understood completely, but studies suggest that when bacteria are treated with zinc nanoparticles, changes taken place in its membrane morphology cause a significant increase in its permeability affecting proper transport through the plasma membrane [68, 69], leaving the bacterial cells incapable of properly regulating transport through the plasma membrane, resulting into cell death [70]. In addition, ZnO nanoparticles could be attributed to the damage of the bacterial cell membrane and extrusion of the cytoplasmic contents thereby resulting in the death of the bacterium [71]. Several investigations have suggested the possible mechanisms involving the interaction of nano-materials with the biological macromolecules. It is believed that microorganisms carry a negative charge while metal oxides carry a positive charge. This creates an ‘‘electromagnetic’’ attraction between the microbe and treated surface [72]. However, to understand the mechanisms of action of these agents, more detailed chemical structure elucidation of the bioactive components followed by therapeutic investigations and toxicological assessment are required.

Techniques for characterization of metallic nanoparticles

The development of materials at nanometric scale is being increased in different fields. The properties of these nanomaterials are critical for the technological revolution worldwide, which mainly depend on the methods of synthesis for the potential applications such as the bactericidal and antifungal effect [73]. For observation of formation and characterization of metallic nanoparticles, several experimental techniques are applied [74-76].

UV-Visible spectroscopy (UV-VIS) is a technique used to quantify the light that is absorbed or scattered by a sample. It is generally recognized that UV–Vis spectra could be used to examine the size and shape controlled nanoparticles in aqueous suspensions [77]. Some of the colloidal metal materials are different under the macroscopic scale and show distinct absorption peaks in the visible region; copper, silver, and gold are metal with prominent absorption peaks. The optical absorption spectra of noble metallic nanoparticles are known to exhibit unique optical properties due to the property of surface plasmon resonance (SPR), which shift to longer wavelengths with increasing the particle size. On the other hand, SPR of a multi-nanoparticle aggregate will be red-shifted to a longer wavelength compared with SPR of the individual particles. SPR is the effect of the oscillation of the conducting electrons aligned in resonance to the wavelength of the irradiated light [78]. The size and shape, the spacing between the metallic nanoparticles and dielectric constant of the medium and surface adsorbed species determine the spectral position of plasmon band absorption as well as its width [79]. According to Mie’s theory, only a single SPR band is expected in the absorption spectra of spherical nanoparticles, whereas anisotropic particles could give rise to two or more SPR bands depending on the shape of the particles. The number of SPR peaks increases as the symmetry of the nanoparticle decreases [80-82]. The color appearing is due to excitation of surface plasmons in metallic nanoparticles [83]. Formation of MNPs by using plant extracts as a result of the reduction of the metal ions to metal is followed by color change and thus UV-Vis spectrum [84, 85].

Transmission Electron Microscopy Analysis (TEM) is the most common characterization technique to determine the size, shape and size distribution of the MNPs.

Scanning Electron Microscopy (SEM)

The SEM image provided insight into the
morphology and size of the synthesized nanoparticles.

X-Ray Diffraction (XRD)

The crystal structure and average particle size of the nanoparticles were analyzed by XRD system [86]. Generally, the narrow and strong diffraction peaks indicate the well crystalline nature of synthesized nanoparticles. The mean size of nanocrystals was measured from the broadening of the diffraction peaks corresponding to the most intensive reflections according to the JCPDS (Joint Committee on Powder Diffraction Standards) database. Scherrer equation [87] was used to determine the crystallite size from XRD diffraction pattern measured for nanoparticles:

d = Kλ/ B cosθ


d– the average dimension of crystallites in nanometers,

λ – the wavelength of the X-ray radiation

K– the Scherrer constant (shape factor, usually taken as 0.94),

B– the line width at half-maximum height (FWHM) in radians,

θ– the Bragg angle, (the position of the diffraction peak maximum).

Energy Dispersive X-Ray Spectroscopy (EDS or EDX)

EDX spectroscopy is used for identification, purity and the elemental composition of the synthesized nanoparticles.

Scanning Electron Microscopy (SEM)–Energy Dispersive X-RAY Spectroscopy (EDX)

The morphology and chemical composition of the synthesized nanoparticles were examined by scanning electron microscopy (SEM) equipped with an energy-dispersive X-ray spectrometer (EDX or EDS). The main use of high-resolution EDS/SEM (∼100 A˚) is the ability to obtain three-dimensional images with large depth fields by a simple sample preparation [88].

Dynamic Light Scattering (DLS) Analysis

The particle size distribution and zeta potential analysis of NPs were evaluated via dynamic light scattering (DLS) and zeta potential was analyzed using Malvern Zetasizer Nano range instrument. The particle size distribution spectra of the metallic nanoparticles were recorded as diameter (nm) versus frequency (%/nm) spectra with diameter (nm) on x-axis and frequency (%/ nm) on y-axis. The zeta potential spectra of the metallic nanoparticles were recorded zeta potential versus intensity spectra; zeta potential (mV) on x-axis and intensity (a.u) on y-axis. Zeta potential values reveal details about the surface charge and predicting the long-term stability of the dispersion of synthesized metallic nanoparticles [89]. Nanoparticles with zeta potential values greater than +25 mV or less than -25 mV typically have high degrees of stability. If the hydrosols have a large negative or positive zeta potential then they will tend to repel each other and there will be no tendency of the particles to agglomerate. On the other hand, the particles have low zeta potential values then there will be no force to prevent the particles coming together and flocculating [90]. The zeta potential of nanoparticles strongly depends on pH and electrolyte concentration of the dispersion [91]. The particle size and zeta potential values of the synthesized AuNPs using mushroom (Agaricus bisporus) extract were 32.1 nm, –45.8 mV respectively. Zeta potential measurements reveal the NPs are highly stable and have an average surface charge of –45.8 mV. As mentioned, the higher zeta-potential value is a key parameter to maintain the stability of suspension through the electrostatic repulsion between particles, which results in a high stability of suspension [92].

Fourier Transform Infrared (FTIR) Spectroscopy

Fourier transform infrared (FTIR) spectroscopy, 
used to evaluate chemical bonds in surface atoms and functional atoms on the surface of nanoparticles, can be used to characterize physical properties of nanomaterials and their functions [93]. In the biosynthesis of MNPs using plant extracts, FTIR spectroscopic measurements were carried out to identify the possible biomolecules in extracts responsible for capping leading to efficient stabilization of MNPs. For instance, FT-IR spectra of as synthesized CuNPs (biocapped) [94] shows major peaks at 2362 cm-1 and 1382 cm-1 corresponding to aldehydic C__H stretching and C__N stretching vibration of the aromatic amine, respectively. These two peaks are absent when CuNPs washed in acetone/methanol/water in the ultrasonic bath. However, the remaining broadband around 3413 cm-1 in both cases indicates the presence of O__H stretching corresponding to poly phenols (flavonoids) present in the plant extract [95]. Also, the bands at 1714 cm-1 and 1108 cm-1 ascribed to C=O stretching and C__O bending indicate the presence of flavonoids and terpenoids which may be responsible for the reduction/stabilization process (Fig. 3) [96].

Synthesis of Nanoparticles Using Plant Extracts

Generally, nanoparticles are prepared by a variety of chemical and physical methods which are not environmentally friendly. Green synthesis of MNPs is an economical, eco-friendly and simple method in the synthesis route [32, 97-99]. A number of bio-molecules act as reducing and protecting agents in the green synthesis of MNPs. Green/biosynthesis of MNPs were performed by using bacteria, fungi and plant extract [100-102]. Green synthesis appears to be a cost-effective alternative to conventional physical and chemical method of MNPs synthesis and would be suitable for developing a biological process for large-scale production. Nowadays, plant extracts act as reducing and capping agents for the synthesis of nanoparticles, which is more advantageous than chemical, microbial synthesis [103-107].

The rate of nanoparticle growth depends upon various variables, including the concentration of metal ions, amount of plant extract, pH and temperature [108]. Time is also a key parameter in the synthesis of nanoparticles. The availability of an enormous number of nuclei at a given time resulted in decreasing the size of nanoparticles, because smaller metal nuclei grow and use metal ions at the same time [109].

Vilchis-Nestor et al. [110] reported the synthesis of gold nanoparticles and silver nanostructures by using green tea (Camellia sinensis) extract in aqueous solution at ambient conditions. They also investigated the control of size, morphology, and optical properties of the nanostructures and reported initial concentrations of metal ions and plant extract as controlling factors. It was investigated that when the amount of C. sinensis extract is increased, the resulted nanoparticles are slightly bigger and more spherical. In another study, the amount of plant material was found to play a critical role in controlling the size and size disparity of metallic nanoparticles. Accordingly, smaller metallic nanoparticles and narrow size distribution occur when more plant extract is added in the reaction mediumThe particle size distribution varied with the variation in the dosage of C. zeylanicum bark extract. The number of particles increased with increasing dosage due to the variation in the number of reductive biomolecules [111].

As metal particles are generated in the aqueous phase, they are unstable by nature, and these metal atoms tend to agglomerate to decrease the total surface energy. In addition, some metals serve as nuclei for others to grow on. This agglomeration, which can be caused by attractive van der Waals forces between crystals, should be repressed to limit the final particle size at the nanometric scale [109]. The plant extract serves as a reducing and dispersing agent to separate metal ions from each other and hence provides better size control of nanoparticles. It also remains on the metal NPs as the capping agent and improves the biological activity. The plant extracts/bioextract often contains metabolites such as flavonoids, proteins, terpenoids, and polyphenols. These biomolecules not only act as reducing agents for metal ion reduction but also (remains on the metallic NPs) as the capping agents which helps to minimize the agglomeration of NPs thereby controlling the morphology and also helping to protect/stabilize the NPs, thus improving the biological potential. G. Sangeetha et al. [112] reported the formation of ZnONPsin the biosynthesis procedures by aloe barbadensis miller leaf extract. The overall observation proved the existence of some phenolic compounds, terpenoids or proteins that were bound to the surface of ZnO nanoparticles. The stability of ZnO nanoparticle could be due to the free amino and carboxylic groups interacted with the zinc surface. The bonds of functional groups such as –CO–C–, –C–O– and –C=C– were derived from heterocyclic compounds and the amide bands derived from the proteins were present in the leaf extract and were the capping ligands of the nanoparticle [113-115]. Moreover, the proteins presented in the medium prevented agglomeration and aided the stabilization by forming a coat, covering the metallic nanoparticles.

In green synthesis of CuNPs, biomolecules found in the Thymus vulgaris L. leaf extract induce the reduction of Cu2+ ions from CuSO4.5H2O to copper nanoparticles [116] or in another synthesis, the plant extract of Nephelium lappaceum acts as ligation, and the aromatic hydroxyl group present in polyphenolic ellagic acid ligate with zinc ions to form zinc-ellagate complex (pH 5–7). Calcification of this complex at 450 °C in static air leads to the formation of ZnO NPs [117]. (Scheme 1)

In some of the studies, the influence of pH on the biosynthesis of NPs has been investigated. It was suggested that different values of pH affect nanoparticle size and shape. In the synthesis of silver and gold nanoparticles by fruit extract of Tanacetum vulgare, larger particle size could be achieved by decreasing the pH [89]. In another study, Dwivedi and Gopal [118] determined that nanoparticles are more stable when exposed to higher pH conditions. Results of studies in the biosynthesis of AgNPs indicate that the size of nanoparticles decreases when pH increases. At lower pH, aggregation exceeds over nucleation to form large particles. Whereas, at higher pH, more functional groups are available for binding to silver leading to the synthesis of stable, small-sized nanoparticles. However, as stated by Gan and Li [119], plant extracts which can produce a large number of stable nanoparticles over wide pH range can be more suitable for application in which there is a change in environmental PH. However, previous studies have indicated that neutral pH is optimal for the synthesis of AgNPs . At this pH, little or no assembly of AgNPs into the particles of suitable size and shape occurs [120]. In another study, nano-crystalline palladium particles (10–15 nm) have been synthesized using Curcuma longa tuber extract as biomaterial that pH and temperature have no major effect on size and shape of the nanoparticles.

Investigations showed that the rate of synthesis of the nanoparticles was related to the reaction and incubation temperature, and an increase in temperature levels leads to nanoparticle growth at a faster rate and reducing their average particle size. The reason for a decrease in particle size with temperature can be discussed as follows. As the reaction temperature increases, the reaction rate increases and thus most metal ions are consumed in the formation of nuclei, stopping the secondary reduction process on the surface of the preformed nuclei. For instance, synthesis rate and final conversion to silver nanoparticles became faster when reaction temperature increased. However, the average particle sizes produced by D. kaki leaf broth decreased from 50 nm to 16 nm when temperature increased from 25 C to 95 C [84].

Table 1 summarizes the important examples of nanoparticle biosynthesis using various plant extracts. Moreover, some important features of the nanoparticles including size and morphology and a number of parameters such as temperature and time are mentioned.


It is known that green synthesis of MNPs is much safer and environmentally friendly compared to chemical and physical synthesis. The methodology employed by using plant extract is very simple, easy to perform, inexpensive, high efficient and eco-friendly. Plant extracts contain diverse chemical compounds such as proteins, carbohydrates, alkaloids, tannins, phenolics, oils, and saponins which have medicinal value and the same can act as reducing and capping agents for MNP synthesis.

The shape, size, and size distribution of MNPs can be controlled by optimization of reaction conditions, such as temperature, pH, and amount of plant material suggesting that extract can be used as both reducing and stabilizing agent for the preparation of MNPs. With the help of more detailed experimentation on reaction parameters such as pH, temperature, ratio and concentration of plant extract to the metal salt, it will be possible to optimize production process to obtain a large amount of stable, and small size MNPs.

Phytosynthesized MNPs have many applications such as antimicrobial, biomedical,
agriculture, bioinsecticides, catalyst, biosensor, etc. The antibacterial activities were inversely proportional to the average nanoparticle sizes. The studies indicated that biologically synthesized metallic nanoparticles using plant extract have higher antibacterial activity than metallic nanoparticles chemically synthesized. Future prospect of plant-mediated nanoparticle synthesis includes an extension of laboratory-based work to industrial scale, elucidation of phytochemicals involved in the synthesis of nanoparticles using bioinformatics tools and deriving the exact mechanism involved in inhibition of pathogenic bacteria.


The authors declare that there is no conflict of interests regarding the publication of this manuscript.



    1. Nanoscience and Nanotechnologies: The Royal Society & The Royal Academy of Engineering; 2004.
    2. Abou El-Nour KMM, Eftaiha Aa, Al-Warthan A, Ammar RAA. Synthesis and applications of silver nanoparticles. Arabian Journal of Chemistry. 2010;3(3):135-40.
    3. Mohanpuria P, Rana NK, Yadav SK. Biosynthesis of nanoparticles: technological concepts and future applications. Journal of Nanoparticle Research. 2007;10(3):507-17.
    4. van Dijken A, Meulenkamp EA, Vanmaekelbergh D, Meijerink A. Identification of the transition responsible for the visible emission in ZnO using quantum size effects. Journal of Luminescence. 2000;90(3-4):123-8.
    5. Moritz M, Geszke-Moritz M. Zastosowanie nanomateriałów w wykrywaniu i usuwaniu zanieczyszczeń środowiska. Przemysł Chemiczny. 2012;91:2375-81.
    6. Zhao G, Stevens JSE. Biometals. 1998;11(1):27-32.
    7. Królikowska A, Kudelski A, Michota A, Bukowska J. SERS studies on the structure of thioglycolic acid monolayers on silver and gold. Surface Science. 2003;532-535:227-32.
    8. Crabtree JH, Burchette RJ, Siddiqi RA, Huen IT, Hadnott LL, Fishman A. The efficacy of silver-ion implanted catheters in reducing peritoneal dialysis-related infections. Peritoneal Dialysis International. 2003;23(4):368-74.
    9. Catauro M, Raucci MG, de Gaetano F, Marotta A. Antibacterial and bioactive silver-containing Na2O·CaO·2SiO2glass prepared by sol–gel method. Journal of Materials Science: Materials in Medicine. 2004;15(7):831-7.
    10. Fardood ST, Ramazani A, Moradi S. Green synthesis of Ni–Cu–Mg ferrite nanoparticles using tragacanth gum and their use as an efficient catalyst for the synthesis of polyhydroquinoline derivatives. Journal of Sol-Gel Science and Technology. 2017;82(2):432-9.
    11. Taghavi Fardood S, Ramazani A, Golfar Z, Joo SW. Green synthesis of Ni-Cu-Zn ferrite nanoparticles using tragacanth gum and their use as an efficient catalyst for the synthesis of polyhydroquinoline derivatives. Applied Organometallic Chemistry. 2017;31(12):e3823.
    12. Arvizo RR, Bhattacharyya S, Kudgus RA, Giri K, Bhattacharya R, Mukherjee P. Intrinsic therapeutic applications of noble metal nanoparticles: past, present and future. Chemical Society Reviews. 2012;41(7):2943.
    13. Beitollai H, Zaimbashi R. A New Sensor Based on Graphite Screen Printed Electrode Modified With Cu-Nanocomplex for Determination of Paracetamol. Nanochemistry Research. 2017; 2 (1): 151-8.
    14. Issa B, Obaidat I, Albiss B, Haik Y. Magnetic Nanoparticles: Surface Effects and Properties Related to Biomedicine Applications. International Journal of Molecular Sciences. 2013;14(11):21266-305.
    15. Rao CNR, Ramakrishna Matte HSS, Voggu R, Govindaraj A. Recent progress in the synthesis of inorganic nanoparticles. Dalton Transactions. 2012;41(17):5089.
    16. Pradeep T. Nano: The Essentials: McGraw-Hill Education; 2008.
    17. Zeiri Y, Elia P, Zach R, Hazan S, Kolusheva S, Porat Ze. Green synthesis of gold nanoparticles using plant extracts as reducing agents. International Journal of Nanomedicine. 2014:4007.
    18. Rai M, Yadav A, Gade A. Silver nanoparticles as a new generation of antimicrobials. Biotechnology Advances. 2009;27(1):76-83.
    19. Khodashenas B, Ghorbani HR. Synthesis of copper nanoparticles : An overview of the various methods. Korean Journal of Chemical Engineering. 2014;31(7):1105-9.
    20. Li X-q, Zhang W-x. Iron Nanoparticles:  the Core−Shell Structure and Unique Properties for Ni(II) Sequestration. Langmuir. 2006;22(10):4638-42.
    21. Li Y, Duan X, Qian Y, Yang L, Liao H. Nanocrystalline Silver Particles: Synthesis, Agglomeration, and Sputtering Induced by Electron Beam. Journal of Colloid and Interface Science. 1999;209(2):347-9.
    22. Maribel G. Guzmán JD, Stephan Godet. Synthesis of silver nanoparticles by chemical reduction method and their antibacterial activity. International Journal of Chemical and Biomolecular Engineering. 2009; 2: 104.
    23. Rodríguez-Sánchez L, Blanco MC, López-Quintela MA. Electrochemical Synthesis of Silver Nanoparticles. The Journal of Physical Chemistry B. 2000;104(41):9683-8.
    24. Sharma VK, Yngard RA, Lin Y. Silver nanoparticles: Green synthesis and their antimicrobial activities. Advances in Colloid and Interface Science. 2009;145(1-2):83-96.
    25. Tan Y, Dai X, Li Y, Zhu D. Preparation of gold, platinum, palladium and silver nanoparticles by the reduction of their salts with a weak reductant–potassium bitartrate. Journal of Materials Chemistry. 2003;13(5):1069-75.
    26. Mallick K, Witcomb MJ, Scurrell MS. Polymer stabilized silver nanoparticles: A photochemical synthesis route. Journal of Materials Science. 2004;39(14):4459-63.
    27. Osaka T, Matsunaga T, Nakanishi T, Arakaki A, Niwa D, Iida H. Synthesis of magnetic nanoparticles and their application to bioassays. Analytical and Bioanalytical Chemistry. 2006;384(3):593-600.
    28. Sun S, Zeng H, Robinson DB, Raoux S, Rice PM, Wang SX, et al. Monodisperse MFe2O4(M = Fe, Co, Mn) Nanoparticles. Journal of the American Chemical Society. 2004;126(1):273-9.
    29. Peng S, Wang C, Xie J, Sun S. Synthesis and Stabilization of Monodisperse Fe Nanoparticles. Journal of the American Chemical Society. 2006;128(33):10676-7.
    30. Raspolli Galletti AM, Antonetti C, Marracci M, Piccinelli F, Tellini B. Novel microwave-synthesis of Cu nanoparticles in the absence of any stabilizing agent and their antibacterial and antistatic applications. Applied Surface Science. 2013;280:610-8.
    31. Roopan SM, Rohit, Madhumitha G, Rahuman AA, Kamaraj C, Bharathi A, et al. Low-cost and eco-friendly phyto-synthesis of silver nanoparticles using Cocos nucifera coir extract and its larvicidal activity. Industrial Crops and Products. 2013;43:631-5.
    32. Roopan SM, Bharathi A, Prabhakarn A, Abdul Rahuman A, Velayutham K, Rajakumar G, et al. Efficient phyto-synthesis and structural characterization of rutile TiO2 nanoparticles using Annona squamosa peel extract. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2012;98:86-90.
    33. Roopan S, Nawaz Khan F. ZnO nanoparticles in the synthesis of AB ring core of camptothecin. Chemical Papers. 2010;64(6).
    34. Kalimuthu K, Suresh Babu R, Venkataraman D, Bilal M, Gurunathan S. Biosynthesis of silver nanocrystals by Bacillus licheniformis. Colloids and Surfaces B: Biointerfaces. 2008;65(1):150-3.
    35. Taghavi Fardood S, Ramazani A. Green Synthesis and Characterization of Copper Oxide Nanoparticles Using Coffee Powder Extract. Journal of Nanostructures. 2016;6(2):167-71.
    36. Taghavi Fardood S, Ramazani A, Moradi S, Azimzadeh Asiabi P. Green synthesis of zinc oxide nanoparticles using arabic gum and photocatalytic degradation of direct blue 129 dye under visible light. Journal of Materials Science: Materials in Electronics. 2017;28(18):13596-601.
    37. Fardood ST, Ramazani A, Moradi S. A Novel Green Synthesis of Nickel Oxide Nanoparticles Using Arabic Gum. Chemistry Journal of Moldova. 2017;12(1):115-8.
    38. Ali Ramazani STF, Zahra Hosseinzadeh, Fariba Sadri, Sang Woo Joo. Green synthesis of magnetic copper ferrite nanoparticles using tragacanth gum as a biotemplate and their catalytic activity for the oxidation of alcohols. Iranian Journal of Catalysis. 2017;7:181-5.
    39. Taghavi Fardood S, Ramazani A, Golfar Z, Joo SW. Green synthesis of Ni-Cu-Zn ferrite nanoparticles using tragacanth gum and their use as an efficient catalyst for the synthesis of polyhydroquinoline derivatives. Applied Organometallic Chemistry. 2017;31(12):e3823.
    40. Kowshik M, Ashtaputre S, Kharrazi S, Vogel W, Urban J, Kulkarni SK, et al. Extracellular synthesis of silver nanoparticles by a silver-tolerant yeast strain MKY3. Nanotechnology. 2002;14(1):95-100.
    41. Rajakumar G, Rahuman AA, Roopan SM, Khanna VG, Elango G, Kamaraj C, et al. Fungus-mediated biosynthesis and characterization of TiO2 nanoparticles and their activity against pathogenic bacteria. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2012;91:23-9.
    42. Shahverdi AR, Minaeian S, Shahverdi HR, Jamalifar H, Nohi A-A. Rapid synthesis of silver nanoparticles using culture supernatants of Enterobacteria: A novel biological approach. Process Biochemistry. 2007;42(5):919-23.
    43. Ankamwar B, Chaudhary M, Sastry M. Gold Nanotriangles Biologically Synthesized using Tamarind Leaf Extract and Potential Application in Vapor Sensing. Synthesis and Reactivity in Inorganic, Metal-Organic, and Nano-Metal Chemistry. 2005;35(1):19-26.
    44. Goodsell DS. Bionanotechnology: John Wiley & Sons, Inc.; 2004 2004/01/02.
    45. Abdel-Halim ES, El-Rafie MH, Al-Deyab SS. Polyacrylamide/guar gum graft copolymer for preparation of silver nanoparticles. Carbohydrate Polymers. 2011;85(3):692-7.
    46. Ledwith DM, Whelan AM, Kelly JM. A rapid, straight-forward method for controlling the morphology of stable silver nanoparticles. Journal of Materials Chemistry. 2007;17(23):2459.
    47. Shankar SS, Rai A, Ahmad A, Sastry M. Controlling the Optical Properties of Lemongrass Extract Synthesized Gold Nanotriangles and Potential Application in Infrared-Absorbing Optical Coatings. Chemistry of Materials. 2005;17(3):566-72.
    48. Morones JR, Elechiguerra JL, Camacho A, Holt K, Kouri JB, Ramírez JT, et al. The bactericidal effect of silver nanoparticles. Nanotechnology. 2005;16(10):2346-53.
    49. Souli M, Galani I, Plachouras D, Panagea T, Armaganidis A, Petrikkos G, et al. Antimicrobial activity of copper surfaces against carbapenemase-producing contemporary Gram-negative clinical isolates. Journal of Antimicrobial Chemotherapy. 2012;68(4):852-7.
    50. He YT, Wan J, Tokunaga T. Kinetic stability of hematite nanoparticles: the effect of particle sizes. Journal of Nanoparticle Research. 2007;10(2):321-32.
    51. Moghaddam AB, Nazari T, Badraghi J, Kazemzad M. Synthesis of ZnO nanoparticles and electrodeposition of polypyrrole/ZnO nanocomposite film. Int J Electrochem Sci. 2009;4(2):247-57.
    52. Kirchner C, Liedl T, Kudera S, Pellegrino T, Muñoz Javier A, Gaub HE, et al. Cytotoxicity of Colloidal CdSe and CdSe/ZnS Nanoparticles. Nano Letters. 2005;5(2):331-8.
    53. Lokina S, Narayanan V. Antimicrobial and anticancer activity of gold nanoparticles synthesized from grapes fruit extract. Chemical Science Transactions. 2013;2(S1):S105-S10.
    54. Rai MK, Deshmukh SD, Ingle AP, Gade AK. Silver nanoparticles: the powerful nanoweapon against multidrug-resistant bacteria. Journal of Applied Microbiology. 2012;112(5):841-52.
    55. Luo Z, Wu Q, Zhang M, Li P, Ding Y. Cooperative antimicrobial activity of CdTe quantum dots with rocephin and fluorescence monitoring for Escherichia coli. Journal of Colloid and Interface Science. 2011;362(1):100-6.
    56. Jan T, Iqbal J, Ismail M, Mahmood A. Synthesis of highly efficient antibacterial agent Ag doped ZnO nanorods: Structural, Raman and optical properties. Journal of Applied Physics. 2014;115(15):154308.
    57. Huh AJ, Kwon YJ. “Nanoantibiotics”: A new paradigm for treating infectious diseases using nanomaterials in the antibiotics resistant era. Journal of Controlled Release. 2011;156(2):128-45.
    58. Feris K, Otto C, Tinker J, Wingett D, Punnoose A, Thurber A, et al. Electrostatic Interactions Affect Nanoparticle-Mediated Toxicity to Gram-Negative BacteriumPseudomonas aeruginosaPAO1. Langmuir. 2010;26(6):4429-36.
    59. Yamamoto O. Influence of particle size on the antibacterial activity of zinc oxide. International Journal of Inorganic Materials. 2001;3(7):643-6.
    60. Padmavathy N, Vijayaraghavan R. Enhanced bioactivity of ZnO nanoparticles—an antimicrobial study. Science and Technology of Advanced Materials. 2008;9(3):035004.
    61. Shah AH, Manikandan E, Basheer Ahamed M, Ahmad Mir D, Ahmad Mir S. Antibacterial and Blue shift investigations in sol–gel synthesized CrxZn1−xO Nanostructures. Journal of Luminescence. 2014;145:944-50.
    62. Yıldırım ÖA, Unalan HE, Durucan C. Highly Efficient Room Temperature Synthesis of Silver-Doped Zinc Oxide (ZnO:Ag) Nanoparticles: Structural, Optical, and Photocatalytic Properties. Journal of the American Ceramic Society. 2013;96(3):766-73.
    63. Zhao J, Wang L, Yan X, Yang Y, Lei Y, Zhou J, et al. Structure and photocatalytic activity of Ni-doped ZnO nanorods. Materials Research Bulletin. 2011;46(8):1207-10.
    64. Priester JH, Stoimenov PK, Mielke RE, Webb SM, Ehrhardt C, Zhang JP, et al. Effects of Soluble Cadmium Salts Versus CdSe Quantum Dots on the Growth of PlanktonicPseudomonas aeruginosa. Environmental Science & Technology. 2009;43(7):2589-94.
    65. Zain NM, Stapley AGF, Shama G. Green synthesis of silver and copper nanoparticles using ascorbic acid and chitosan for antimicrobial applications. Carbohydrate Polymers. 2014;112:195-202.
    66. Santo CE, Quaranta D, Grass G. Antimicrobial metallic copper surfaces killStaphylococcus haemolyticusvia membrane damage. MicrobiologyOpen. 2012;1(1):46-52.
    67. Ren G, Hu D, Cheng EWC, Vargas-Reus MA, Reip P, Allaker RP. Characterisation of copper oxide nanoparticles for antimicrobial applications. International Journal of Antimicrobial Agents. 2009;33(6):587-90.
    68. Auffan M, Rose J, Bottero J-Y, Lowry GV, Jolivet J-P, Wiesner MR. Towards a definition of inorganic nanoparticles from an environmental, health and safety perspective. Nature Nanotechnology. 2009;4(10):634-41.
    69. Brayner R, Ferrari-Iliou R, Brivois N, Djediat S, Benedetti MF, Fiévet F. Toxicological Impact Studies Based onEscherichiacoliBacteria in Ultrafine ZnO Nanoparticles Colloidal Medium. Nano Letters. 2006;6(4):866-70.
    70. Raffi M, Hussain F, Bhatti T, Akhter J, Hameed A, Hasan M. Antibacterial characterization of silver nanoparticles against E. coli ATCC-15224. Journal of materials science and technology. 2008;24(2):192-6.
    71. Divyapriya S, Sowmia C, Sasikala S. Synthesis of zinc oxide nanoparticles and antimicrobial activity of Murraya Koenigii. World J Pharm Pharm Sci. 2014;3(12):1635-45.
    72. Rezaei-Zarchi S, Javed A, Javeed Ghani M, Soufian S, Barzegari Firouzabadi F, Bayanduri Moghaddam A, et al. Comparative study of antimicrobial activities of TiO2 and CdO nanoparticles against the pathogenic strain of Escherichia coli. Iranian Journal of Pathology. 2010;5(2):83-9.
    73. Ruparelia JP, Chatterjee AK, Duttagupta SP, Mukherji S. Strain specificity in antimicrobial activity of silver and copper nanoparticles. Acta Biomaterialia. 2008;4(3):707-16.
    74. Ghatak KL. Techniques and Methods in biology: PHI Learning; 2011.
    75. Sareen K. Instrumental Methods of Environmental Analysis: IVY Publishing House; 2001.
    76. Inorganic Spectroscopy And Related Topics: sarup; 2008.
    77. W RR, R LJ, S KN, D MV, B KS. Phytosynthesis of Silver Nanoparticle Using Gliricidia sepium (Jacq.). Current Nanoscience. 2009;5(1):117-22.
    78. Parameshwaran R, Kalaiselvam S, Jayavel R. Green synthesis of silver nanoparticles using Beta vulgaris: Role of process conditions on size distribution and surface structure. Materials Chemistry and Physics. 2013;140(1):135-47.
    79. Smitha SL, Nissamudeen KM, Philip D, Gopchandran KG. Studies on surface plasmon resonance and photoluminescence of silver nanoparticles. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2008;71(1):186-90.
    80. Sosa IO, Noguez C, Barrera RG. Optical Properties of Metal Nanoparticles with Arbitrary Shapes. The Journal of Physical Chemistry B. 2003;107(26):6269-75.
    81. Caceres A, Lopez BR, Giron MA, Logemann H. Plants used in guatemala for the treatment of dermatophytic infections. 1. Screening for antimycotic activity of 44 plant extracts. Journal of Ethnopharmacology. 1991;31(3):263-76.
    82. Cáceres A, Menéndez H, Méndez E, Cohobón E, Samayoa BE, Jauregui E, et al. Antigonorrhoeal activity of plants used in Guatemala for the treatment of sexually transmitted diseases. Journal of Ethnopharmacology. 1995;48(2):85-8.
    83. Mulvaney P. Surface Plasmon Spectroscopy of Nanosized Metal Particles. Langmuir. 1996;12(3):788-800.
    84. Song JY, Kim BS. Rapid biological synthesis of silver nanoparticles using plant leaf extracts. Bioprocess and Biosystems Engineering. 2008;32(1):79.
    85. Das RK, Borthakur BB, Bora U. Green synthesis of gold nanoparticles using ethanolic leaf extract of Centella asiatica. Materials Letters. 2010;64(13):1445-7.
    86. Cassetta A. X-Ray Diffraction (XRD). In: Drioli E, Giorno L, editors. Encyclopedia of Membranes. Berlin, Heidelberg: Springer Berlin Heidelberg; 2015. p. 1-3.
    87. Holzwarth U, Gibson N. The Scherrer equation versus the’Debye-Scherrer equation’. Nature nanotechnology. 2011;6(9):534.
    88. Gaber M, El-Sayed YS, El-Baradie K, Fahmy RM. Cu(II) complexes of monobasic bi- or tridentate (NO, NNO) azo dye ligands: Synthesis, characterization, and interaction with Cu-nanoparticles. Journal of Molecular Structure. 2013;1032:185-94.
    89. Dubey SP, Lahtinen M, Sillanpää M. Tansy fruit mediated greener synthesis of silver and gold nanoparticles. Process Biochemistry. 2010;45(7):1065-71.
    90. Roy S, Mukherjee T, Chakraborty S, Das TK. Biosynthesis, characterisation & antifungal activity of Silver nanoparticles synthesized by the fungus aspergillus Foetidus mtcc8876. Digest Journal of Nanomaterials and Biostructures. 2013;8(1):197-205.
    91. Sukirtha R, Priyanka KM, Antony JJ, Kamalakkannan S, Thangam R, Gunasekaran P, et al. Cytotoxic effect of Green synthesized silver nanoparticles using Melia azedarach against in vitro HeLa cell lines and lymphoma mice model. Process Biochemistry. 2012;47(2):273-9.
    92. Eskandari-Nojedehi M, Jafarizadeh-Malmiri H, Rahbar-Shahrouzi J. Hydrothermal green synthesis of gold nanoparticles using mushroom (Agaricus bisporus) extract: physico-chemical characteristics and antifungal activity studies. Green Processing and Synthesis. 2018;7(1):38-47.
    93. Morais PC, Santos RL, Pimenta ACM, Azevedo RB, Lima ECD. Preparation and characterization of ultra-stable biocompatible magnetic fluids using citrate-coated cobalt ferrite nanoparticles. Thin Solid Films. 2006;515(1):266-70.
    94. Yallappa S, Manjanna J, Sindhe MA, Satyanarayan ND, Pramod SN, Nagaraja K. Microwave assisted rapid synthesis and biological evaluation of stable copper nanoparticles using T. arjuna bark extract. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2013;110:108-15.
    95. Raghunandan D, Mahesh BD, Basavaraja S, Balaji SD, Manjunath SY, Venkataraman A. Microwave-assisted rapid extracellular synthesis of stable bio-functionalized silver nanoparticles from guava (Psidium guajava) leaf extract. Journal of Nanoparticle Research. 2011;13(5):2021-8.
    96. Mallick K, Witcomb MJ, Scurrell MS. In situ synthesis of copper nanoparticles and poly(o-toluidine): A metal–polymer composite material. European Polymer Journal. 2006;42(3):670-5.
    97. Sondi I, Salopek-Sondi B. Silver nanoparticles as antimicrobial agent: a case study on E. coli as a model for Gram-negative bacteria. Journal of Colloid and Interface Science. 2004;275(1):177-82.
    98. Raffi M, Rumaiz AK, Hasan MM, Shah SI. Studies of the growth parameters for silver nanoparticle synthesis by inert gas condensation. Journal of Materials Research. 2011;22(12):3378-84.
    99. Kumar R, Roopan SM, Prabhakarn A, Khanna VG, Chakroborty S. Agricultural waste Annona squamosa peel extract: Biosynthesis of silver nanoparticles. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2012;90:173-6.
    100. Philip D, Unni C, Aromal SA, Vidhu VK. Murraya Koenigii leaf-assisted rapid green synthesis of silver and gold nanoparticles. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2011;78(2):899-904.
    101. Kaviya S, Santhanalakshmi J, Viswanathan B, Muthumary J, Srinivasan K. Biosynthesis of silver nanoparticles using citrus sinensis peel extract and its antibacterial activity. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2011;79(3):594-8.
    102. Husseiny MI, El-Aziz MA, Badr Y, Mahmoud MA. Biosynthesis of gold nanoparticles using Pseudomonas aeruginosa. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2007;67(3):1003-6.
    103. Smitha SL, Philip D, Gopchandran KG. Green synthesis of gold nanoparticles using Cinnamomum zeylanicum leaf broth. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2009;74(3):735-9.
    104. Bhainsa KC, D’Souza SF. Extracellular biosynthesis of silver nanoparticles using the fungus Aspergillus fumigatus. Colloids and Surfaces B: Biointerfaces. 2006;47(2):160-4.
    105. Ahmad A, Mukherjee P, Senapati S, Mandal D, Khan MI, Kumar R, et al. Extracellular biosynthesis of silver nanoparticles using the fungus Fusarium oxysporum. Colloids and Surfaces B: Biointerfaces. 2003;28(4):313-8.
    106. Sorbiun M, Shayegan Mehr E, Ramazani A, Taghavi Fardood S. Biosynthesis of Ag, ZnO and bimetallic Ag/ZnO alloy nanoparticles by aqueous extract of oak fruit hull (Jaft) and investigation of photocatalytic activity of ZnO and bimetallic Ag/ZnO for degradation of basic violet 3 dye. Journal of Materials Science: Materials in Electronics. 2018;29(4):2806-14.
    107. Shayegan Mehr E, Sorbiun M, Ramazani A, Taghavi Fardood S. Plant-mediated synthesis of zinc oxide and copper oxide nanoparticles by using ferulago angulata (schlecht) boiss extract and comparison of their photocatalytic degradation of Rhodamine B (RhB) under visible light irradiation. Journal of Materials Science: Materials in Electronics. 2018;29(2):1333-40.
    108. Han KN, Kim NS. Challenges and opportunities in direct write technology using nano-metal particles. KONA Powder and Particle Journal. 2009;27:73-83.
    109. Thi My Dung D, Thi Tuyet Thu L, Eric F-B, Mau Chien D. Synthesis and optical properties of copper nanoparticles prepared by a chemical reduction method. Advances in Natural Sciences: Nanoscience and Nanotechnology. 2011;2(1):015009.
    110. Vilchis-Nestor AR, Sánchez-Mendieta V, Camacho-López MA, Gómez-Espinosa RM, Camacho-López MA, Arenas-Alatorre JA. Solventless synthesis and optical properties of Au and Ag nanoparticles using Camellia sinensis extract. Materials Letters. 2008;62(17):3103-5.
    111. Sathishkumar M, Sneha K, Won SW, Cho CW, Kim S, Yun YS. Cinnamon zeylanicum bark extract and powder mediated green synthesis of nano-crystalline silver particles and its bactericidal activity. Colloids and Surfaces B: Biointerfaces. 2009;73(2):332-8.
    112. Sangeetha G, Rajeshwari S, Venckatesh R. Green synthesis of zinc oxide nanoparticles by aloe barbadensis miller leaf extract: Structure and optical properties. Materials Research Bulletin. 2011;46(12):2560-6.
    113. Jiale H, Qingbiao L, Daohua S, Yinghua L, Yuanbo S, Xin Y, et al. Biosynthesis of silver and gold nanoparticles by novel sundried Cinnamomum camphora leaf. Nanotechnology. 2007;18(10):105104.
    114. Sastry M, Ahmad A, Khan MI, Kumar R. Biosynthesis of metal nanoparticles using fungi and actinomycete. Current science. 2003;85(2):162-70.
    115. Sanghi R, Verma P. Biomimetic synthesis and characterisation of protein capped silver nanoparticles. Bioresource Technology. 2009;100(1):501-4.
    116. Issaabadi Z, Nasrollahzadeh M, Sajadi SM. Green synthesis of the copper nanoparticles supported on bentonite and investigation of its catalytic activity. Journal of Cleaner Production. 2017;142:3584-91.
    117. Karnan T, Selvakumar SAS. Biosynthesis of ZnO nanoparticles using rambutan (Nephelium lappaceumL.) peel extract and their photocatalytic activity on methyl orange dye. Journal of Molecular Structure. 2016;1125:358-65.
    118. Dwivedi AD, Gopal K. Biosynthesis of silver and gold nanoparticles using Chenopodium album leaf extract. Colloids and Surfaces A: Physicochemical and Engineering Aspects. 2010;369(1):27-33.
    119. Gan PP, Li SFY. Potential of plant as a biological factory to synthesize gold and silver nanoparticles and their applications. Reviews in Environmental Science and Bio/Technology. 2012;11(2):169-206.
    120. Iravani S, Zolfaghari B. Green synthesis of silver nanoparticles using Pinus eldarica bark extract. BioMed research international. 2013;2013.
    121. Vanathi P, Rajiv P, Narendhran S, Rajeshwari S, Rahman PKSM, Venckatesh R. Biosynthesis and characterization of phyto mediated zinc oxide nanoparticles: A green chemistry approach. Materials Letters. 2014;134:13-5.
    122. Dobrucka R, Długaszewska J. Biosynthesis and antibacterial activity of ZnO nanoparticles using Trifolium pratense flower extract. Saudi Journal of Biological Sciences. 2016;23(4):517-23.
    123. Supraja N, Prasad TNVKV, Krishna TG, David E. Synthesis, characterization, and evaluation of the antimicrobial efficacy of Boswellia ovalifoliolata stem bark-extract-mediated zinc oxide nanoparticles. Applied Nanoscience. 2016;6(4):581-90.
    124. Stan M, Popa A, Toloman D, Silipas T-D, Vodnar DC. Antibacterial and Antioxidant Activities of ZnO Nanoparticles Synthesized Using Extracts of Allium sativum, Rosmarinus officinalis and Ocimum basilicum. Acta Metallurgica Sinica (English Letters). 2016;29(3):228-36.
    125. Jafari A, Pourakbar L, Farhadi K, GHOLIZAD LM, Goosta Y. Biological synthesis of silver nanoparticles and evaluation of antibacterial and antifungal properties of silver and copper nanoparticles. Turkish Journal of Biology. 2015;39(4):556-61.
    126. Lee HJ, Song JY, Kim BS. Biological synthesis of copper nanoparticles using Magnolia kobus leaf extract and their antibacterial activity. Journal of Chemical Technology and Biotechnology. 2013;88(11):1971-7.
    127. Buazar F, Bavi M, Kroushawi F, Halvani M, Khaledi-Nasab A, Hossieni SA. Potato extract as reducing agent and stabiliser in a facile green one-step synthesis of ZnO nanoparticles. Journal of Experimental Nanoscience. 2016;11(3):175-84.
    128. Karimi J, Mohsenzadeh S. Rapid, Green, and Eco-Friendly Biosynthesis of Copper Nanoparticles Using Flower Extract of Aloe Vera. Synthesis and Reactivity in Inorganic, Metal-Organic, and Nano-Metal Chemistry. 2015;45(6):895-8.
    129. Buazar F, Baghlani-Nejazd MH, Badri M, Kashisaz M, Khaledi-Nasab A. 2 Facile one-pot phytosynthesis of magnetic nanoparticles using 3 potato extract and their catalytic activity. Starch/Stärke. 2016;68:1-9.
    130. Song JY, Jang H-K, Kim BS. Biological synthesis of gold nanoparticles using Magnolia kobus and Diopyros kaki leaf extracts. Process Biochemistry. 2009;44(10):1133-8.
    131. Moon SA, Salunke BK, Alkotaini B, Sathiyamoorthi E, Kim BS. Biological synthesis of manganese dioxide nanoparticles by Kalopanax pictus plant extract. IET Nanobiotechnology [Internet]. ٢٠١٥; ٩(٤):[٢٢٠-٥ pp.]. Available from:١٠.١٠٤٩/iet-nbt.٢٠١٤.٠٠٥١.
    132. Song JY, Kwon E-Y, Kim BS. Biological synthesis of platinum nanoparticles using Diopyros kaki leaf extract. Bioprocess and Biosystems Engineering. 2009;33(1):159.
    133. Li C-y, Zhang Z-c, Mao J-y, Shi L-f, Zheng Y, Quan J-l. Preparation of Tradescantia pallida-mediated zinc oxide nanoparticles and their activity against cervical cancer cell lines. Tropical Journal of Pharmaceutical Research. 2017;16(3):494-500.
    134. Song JY, Kwon E-Y, Kim BS. Antibacterial latex foams coated with biologically synthesized silver nanoparticles using Magnolia kobus leaf extract. Korean Journal of Chemical Engineering. 2012;29(12):1771-5.
    135. Salunke BK, Shin J, Sawant SS, Alkotaini B, Lee S, Kim BS. Rapid biological synthesis of silver nanoparticles using Kalopanax pictus plant extract and their antimicrobial activity. Korean Journal of Chemical Engineering. 2014;31(11):2035-40.
    136. Salunke BK, Sawant SS, Kim BS. Potential of Kalopanax septemlobus Leaf Extract in Synthesis of Silver Nanoparticles for Selective Inhibition of Specific Bacterial Strain in Mixed Culture. Applied Biochemistry and Biotechnology. 2014;174(2):587-601.
    137. Borase HP, Patil CD, Salunkhe RB, Suryawanshi RK, Kim BS, Bapat VA, et al. Bio-Functionalized Silver Nanoparticles: a Novel Colorimetric Probe for Cysteine Detection. Applied Biochemistry and Biotechnology. 2015;175(7):3479-93.
    138. Pedrycz W, Bortolan G, Degani R. Classification of electrocardiographic signals: a fuzzy pattern matching approach. Artificial Intelligence in Medicine. 1991;3(4):211-26.
    139. Mariselvam R, Ranjitsingh AJA, Usha Raja Nanthini A, Kalirajan K, Padmalatha C, Mosae Selvakumar P. Green synthesis of silver nanoparticles from the extract of the inflorescence of Cocos nucifera (Family: Arecaceae) for enhanced antibacterial activity. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2014;129:537-41.
    140. Ashokkumar S, Ravi S, Kathiravan V, Velmurugan S. RETRACTED: Synthesis of silver nanoparticles using A. indicum leaf extract and their antibacterial activity. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2015;134:34-9.
    141. Korbekandi H, Chitsazi MR, Asghari G, Bahri Najafi R, Badii A, Iravani S. Green biosynthesis of silver nanoparticles using Quercus brantii (oak) leaves hydroalcoholic extract. Pharmaceutical biology. 2015;53(6):807-12.
    142. Jha AK, Prasad K. Green synthesis of silver nanoparticles using Cycas leaf. International Journal of Green Nanotechnology: Physics and Chemistry. 2010;1(2):P110-P7.
    143. Kaur P, Thakur R, Chaudhury A. Biogenesis of copper nanoparticles using peel extract of Punica granatum and their antimicrobial activity against opportunistic pathogens. Green Chemistry Letters and Reviews. 2016;9(1):33-8.
    144. Ahmad N, Sharma S. Green synthesis of silver nanoparticles using extracts of Ananas comosus. Green and Sustainable Chemistry. 2012;2(04):141.
    145. Sharma G, Sharma A, Bhavesh R, Park J, Ganbold B, Nam J-S, et al. Biomolecule-Mediated Synthesis of Selenium Nanoparticles using Dried Vitis vinifera (Raisin) Extract. Molecules. 2014;19(3):2761.
    146. Rathnasamy R, Thangasamy P, Thangamuthu R, Sampath S, Alagan V. Green synthesis of ZnO nanoparticles using Carica papaya leaf extracts for photocatalytic and photovoltaic applications. Journal of Materials Science: Materials in Electronics. 2017;28(14):10374-81.
    147. Aminuzzaman M, Kei LM, Liang WH, editors. Green synthesis of copper oxide (CuO) nanoparticles using banana peel extract and their photocatalytic activities. AIP Conference Proceedings; 2017: AIP Publishing.
    148. Gallucci MN, Fraire JC, Ferreyra Maillard APV, Páez PL, Aiassa Martínez IM, Pannunzio Miner EV, et al. Silver nanoparticles from leafy green extract of Belgian endive (Cichorium intybus L. var. sativus): Biosynthesis, characterization, and antibacterial activity. Materials Letters. 2017;197:98-101.
    149. Patil MP, Jin X, Simeon NC, Palma J, Kim D, Ngabire D, et al. Anticancer activity of Sasa borealis leaf extract-mediated gold nanoparticles. Artificial Cells, Nanomedicine, and Biotechnology. 2018;46(1):82-8.
    150. Rajan A, Rajan AR, Philip D. Elettaria cardamomum seed mediated rapid synthesis of gold nanoparticles and its biological activities. OpenNano. 2017;2:1-8.
    151. Maruthupandy M, Zuo Y, Chen J-S, Song J-M, Niu H-L, Mao C-J, et al. Synthesis of metal oxide nanoparticles (CuO and ZnO NPs) via biological template and their optical sensor applications. Applied Surface Science. 2017;397:167-74.
    152. Jacob SJP, Prasad VLS, Sivasankar S, Muralidharan P. Biosynthesis of silver nanoparticles using dried fruit extract of Ficus carica - Screening for its anticancer activity and toxicity in animal models. Food and Chemical Toxicology. 2017;109:951-6.