A comparative study of eco-friendly silver nanoparticles synthesis using Prunus domestica plum extract and sodium citrate as reducing agents

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Advanced Powder Technology journal homepage: www.elsevier.com/locate/apt

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Original Research Paper

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A comparative study of eco-friendly silver nanoparticles synthesis using Prunus domestica plum extract and sodium citrate reducing agents

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Seraj Mohaghegh a,b,1, Karim Osouli-Bostanabad a,b,1, Hossein Nazemiyeh a, Yousef Javadzadeh c, Alireza Parvizpur c, Mohammad Barzegar-Jalali d,⇑, Khosro Adibkia a,c,⇑ a

Research Center for Pharmaceutical Nanotechnology, and Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran Students Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran d Pharmaceutical Analysis Research Center, Tabriz University of Medical Sciences, Tabriz, Iran b c

a r t i c l e

i n f o

Article history: Received 28 August 2019 Received in revised form 2 November 2019 Accepted 25 December 2019 Available online xxxx Keywords: Biosynthesis Environmental benign Prunus domestica Silver nanoparticles

a b s t r a c t Through the current comparative study, colloidal silver nanoparticles (AgNPs) synthesized with various morphologies and sizes using Prunus domestica (P-dom) extract and sodium citrate as green and chemical reducing agents, respectively. AgNPs were synthesized employing different concentrations of the reducing agents in an aqueous solution at various pH values (3–10) and temperatures (25–85 °C). The UV–visible absorption spectrum indicated characteristic SPR peaks of AgNPs at 380–450 nm. Fourier transform infrared spectroscopy revealed aqueous-soluble polyols (such as glycosides, phenols, and flavanols) participation in Ag ions reduction to the corresponding AgNPs at various pH values. The crystallinity of AgNPs detected by an X-ray diffractometer. Different morphologies (polygonal, oval, and spherical) of AgNPs with varying pH values were confirmed conducting transmission electron microscopy (TEM). Average particle sizes of 16–50 nm were determined using scanning electron microscopy, TEM, and dynamic light scattering assessments for AgNPs synthesized at various reaction conditions. This study is a demonstration for a facile, cheap, and eco-friendly stimuli-sensitive synthesize of AgNPs. Ó 2020 Published by Elsevier B.V. on behalf of The Society of Powder Technology Japan. All rights reserved.

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1. Introduction

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Around five decades ago, it was proposed that the material properties could be enhanced by miniaturization. In ancient times, nanomaterials were employed with inadequate facilities to probe at the nano-size and limited grasp of nanotechnology. These particles indicate augmented and novel properties based on their unique characteristics such as morphology, size, and specific surface area [1,2]. The utilization of nanoscale structures and materials, commonly in the range of 1–100 nm (nm) with the ability to probe, is an emerging domain of nanotechnology and nanoscience. Nanomaterials may offer solutions to environmental and technological challenges in the fields of catalysis, solar energy conversion, water treatment, and medicine [3,4]. Nanomaterials synthesis can be conducted by numerous physical and chemical approaches

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⇑ Corresponding authors at: Faculty of Pharmacy, Tabriz University of Medical Sciences, Golgasht Street, Daneshgah Ave, Tabriz, Iran (K. Adibkia). E-mail addresses: [email protected] (M. Barzegar-Jalali), [email protected]. ir (K. Adibkia). 1 Both authors equally contributed to this study.

including, but not restricted to hydrothermal, microwave assisted, chemical reduction, heat evaporation, electrochemical reduction, photochemical reduction, and so on [5–10]. While these techniques successfully can be applied to the synthesis of nanomaterials, the increasing demand regards of nanomaterials applications must be accompanied by eco-friendly synthesis approaches. In the worldwide efforts to minimize produced hazardous byproducts or wastes from chemical/physical processes, these procedures are increasingly integrating with the latest evolutions in science and technology. Implementation of these continual procedures should adopt the twelve primary rules of green chemistry [11,12]. These principles are adapted to lead in reducing unsafe product usage and increasing the effectiveness of chemical processes. Therefore, any synthetic method or chemical procedure should obey these principles by employing nontoxic chemicals and environmentally benign solvents. Previous investigations have been concentrated on the significance of metal nanoparticles including palladium (Pd), platinum (Pt) copper (Cu), gold (Au), and silver (Ag) because of their exceptional nanoscale size, physicochemical and surface plasmon properties [1,13,14]. Among these, Ag nanoparticles (AgNPs) show an

https://doi.org/10.1016/j.apt.2019.12.039 0921-8831/Ó 2020 Published by Elsevier B.V. on behalf of The Society of Powder Technology Japan. All rights reserved.

Please cite this article as: S. Mohaghegh, K. Osouli-Bostanabad, H. Nazemiyeh et al., A comparative study of eco-friendly silver nanoparticles synthesis using Prunus domestica plum extract and sodium citrate reducing agents, Advanced Powder Technology, https://doi.org/10.1016/j.apt.2019.12.039

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outstanding and unique size and morphology-dependent thermal, electrical, catalytic, optical, optoelectronic, bio-sensing, anticancer, biological, medicinal, and antiviral characteristics. AgNPs are an exceptional candidate for numerous applications in a various disciplines and fields such as anticancer, drug delivery, dentistry, antioxidant, antimicrobial, cosmetic, water treatment, agriculture, food science, larvicides, clothing, forensic science, waste management, pollution control, chemistry, photovoltaics, and biomedical industry [1,15–17]. The established procedures for AgNPs synthesis use nonenvironment friendly and toxic reagents. Therefore, alternatives to these chemical techniques have been introduced recently, replacing the conventional reducing compounds with natural antioxidant agents. These approaches are known as ‘‘green/ biological synthesis” due to the following principles of green chemistry. As non-pathogenic methods to synthesis AgNPs, employing fruits, plants, and their extracts are straightforward, one-step processes with a higher bio-reduction capability compared to the templates and microorganisms usage [18]. Green reducing agents have various functional groups such as terpenoids, ketones, phenolic compounds, carboxylic acids, aldehydes, alkaloids, amines, and co-enzymes that can be corresponded to a different metal ions reduction to metal atoms [19]. Organic acids, flavones, and quinones are engaged in Ag+ ions reduction immediately to Ag° in a reaction medium [20]. Recognizing the activity of each abovementioned group in the nucleation, reduction, and growth of correspondent metal nanostructures from metal ions would facilitate it to prognosticate the applicability of other new green reducing compounds for the synthesis of metal nanostructures. Furthermore, this study would be necessary for realizing how the morphology and size of the synthesized NPs could be regulated. Many. Different kinds of plants have been successfully applied in the synthesis of biocompatible, and stable bioactive AgNPs with potential applications in therapeutic areas [21,22]. Prunus domestica (P-dom) (plum) usually called European plum, is also a stone fruit harvest having prehistoric derivation and cultivation alike as an apple. P-dom has been discovered at the same time as fig and grapes in archaeological explorations [23] that has ample amounts of antioxidants (i.e., (+)-abscisic acid, chlorogenic acid (5-O-caffeoylquinicacid), (6S,9R)-roseoside, neochlorogenic acid (3-Ocaffeoylquinic acid), cryptochlorogenic acid (4-O-caffeoylquinic acid), (+)-b-D-glucopyranosyl abscisate, and two lignan glucosides). Furthermore, it contains abundant of bioactive agents such as anthocyanins, flavonoids (kaempferol, quercetin, and myricetin), carbohydrates (sucrose, fructose, sorbitol, glucose), phenols, organic acids (malic acid, citric acid), minerals (potassium, sodium, calcium, magnesium, zinc, iron) and vitamins (c-tocopherol, a-tocopherol, b-carotene). In turn, provide it therapeutic merits in natural drugs syntheses and being used in disorders related to the female reproductive cycle [24–27]. It has been presumed that P-dom could be used as a therapeutic compound to decrease chronic health ailments risk like cardiovascular disease and cancer [28]. Studies have shown that the properties of as-synthesized AgNPs are primarily connected to their preparation techniques. The structure, size and corresponding chemical, physical, and biological features are highly dependent on the synthetic procedure [29–31]. Therefore, the design of a synthesis process in which the stability, morphology, size, and properties are controlled is a crucial aspect of nanotechnology. The reaction mechanism realization makes it feasible to understand how the morphology and size of NPs can be tuned and controlled. Appropriate characterizations within the reaction can also be employed to explore the occurring reaction kinetics. For example, in the majority of green synthesis methods of metal NPs, at the start of the reaction NPs nucleation and growth

are slow, but are quickened once the first seeds form that fit to the sigmoidal kinetic [32]. To the best of our knowledge, there has been no effort to use Prunus domestica (P-dom) fruit extract as a reducing and capping agent to prepare AgNPs. Since we further concentrated on the employing of P-dom for the synthesis of AgNPs with controllable morphology and size. The main aims of the current study were: (I) to synthesis AgNPs using P-dom extract via green nanotechnology process, and (II) employing a wet chemical method (citrate as a reducing agent); (III) to apply various temperatures, pH values, and concentrations of the reducing agent for optimizing the synthesizing procedure of AgNPs; and (IV) to assess the synthesized nanoparticles physicochemical characteristics. In a word, this study would be worthwhile in providing a comparison between two methods of synthesizing AgNPs (i.e., green and wet chemical approaches) as well as introducing a new reducing agent for the synthesis and stabilization of AgNPs.

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2. Materials and methods

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2.1. Materials and chemicals

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All the chemicals and solvents employed in the current study were of analytical grade and used without further purification supplied by Sigma Aldrich. Ultrapure Milli-Q water (at ambient temperature with the specific resistivity of 18.2 MX.cm) was utilized as a solvent in all experiments.

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2.2. Preparation of Prunus domestica (plum) extract

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An appropriate amount of fruits was thoroughly washed with distilled water to remove any impurities and dust. Fresh fruits of P-dom (2000 g) were chopped and added in ethanol (70%) with a ratio of 1:3 w/v (P-dom: ethanol) at ambient temperature for 72 h under shaking at 300 rpm. The extracts were removed and filtered using a filter paper of Whatman No. 1. Subsequently, the filtered extract was condensed by removing ethanol under reduced pressure at 40 °C, employing a rotary evaporator (100 rpm, low pressure) (Heidolph Persia, Iran). The condensed and dried crude extract was meticulously weighed and kept in a refrigerator at 4 °C for further study.

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2.3. Green synthesis procedure

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Through the current study, to investigate and optimize the effectiveness of synthesizing methods with biological and chemical reducing agents at different pH values (i.e. 3, 5, 7 and 10), various experiments were carried out for a predetermined time (20 min) using different concentrations of the extract (1, 2.5, 5 and 7.5 % w/v) at 25, 45, 60 and 85 °C. Briefly, 10 ml of AgNO3 solution (1 mM) equilibrated at the desired temperature before adding the solution of P-dom extract. To maintain the pH at the appropriate value, hydrochloric acid, and sodium hydroxide 0.1 N solutions were used (the pH of the prepared AgNO3 solution was 5). After adjusting the pH, 1 ml of P-dom extract was added dropwise to the aforementioned solution under continues agitation, and the solution was left for 20 min in this condition to complete the reduction of Ag+ ions to Ago. The particles formation was tracked visually by the color change of the solution from yellowish to colloidal brown signifying the formation of Ag particles. The acquired suspension was centrifuged at 13,000 rpm for 15 min (three times) and used for further analyses and applications.

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2.4. Wet chemical synthesis procedure

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Sodium citrate as the common reducing compound of the wet chemical technique was used to synthesize AgNPs and compared with the conducted bio-synthesizing method. In this regard, a solution of sodium citrate 1% w/v was employed as reducing agent, where to prepare it, 1 g of tri-sodium citrate was dissolved in 100 ml of distillated water and the experiments for synthesizing Ag particles were done at different pH values (i.e., 3, 5, 7 and 10). Various experiments were carried out for a predetermined time (20 min) at 85 °C.

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2.5. Characterization

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2.5.1. UV–vis study The appearance of excitation spectra (localized surface plasmon resonance) were recorded with a scan rate of 60 nm/min conducting Shimadzu UV–visible spectrometry (Shimadzu Spectrophotometer, Japan) in the range of 300–700 nm. The synthesized colloidal solutions were poured in quartz cuvettes with a length path of 1 cm and diluted using milli-Q water once the sample had a too high optical density (i.e., the absorbance was higher than 2.5 in the range of 300–700 nm).

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2.5.2. Fourier-transform infrared analysis Fourier-transform infrared (FT-IR) spectra of as synthesized particles using green synthesis were recorded on Shimadzu FTIR spectrophotometer (Shimadzu 43000, Kyoto, Japan). For doing the survey, the synthesized particles were compacted in a disc shape by KBr disk method and studied at a resolution of 2 cm
1 with an average spectra of 32 scans in the scanning range of 4000–500 cm
1. 2.5.3. X-ray diffraction examination and Energy-dispersive X-ray spectroscopy The X-ray diffraction patterns of as synthesized particles were measured conducting X-ray diffractometer D 5000 (Siemens, Munich, Germany) at a step size of 0.02° in 2h angle range of 10– 80° with a scanning rate of 0.6°/min. The operational parameters were Cu Ka radiation (k = 1.5405 Å) at 40 kV, 30 mA. The elemental composition of the bio-synthesized silver nanoparticles (P-dom-AgNPs) was determined by EDX attached to a field emission scanning electron microscope (FE-SEM, MIRA3, Tescan Co., Brno, Czech) at an operational 20 kV condition. 2.5.4. Field emission scanning, transmission electron microscopies and dynamic light scattering assessments The synthesized samples morphology and size were assessed employing a field emission scanning electron microscope (FE-SEM, MIRA3, Tescan Co., Brno, Czech) at an operational 20 kV condition and Transmission electron microscopy (TEM, 20 G2 Tecnai, 80 kV). The synthesized samples were coated with a thin gold layer (about 150 Å in thickness) using gold sputtering apparatus (Emitech K550, Kent, UK) before evaluation by FE-SEM. Samples for TEM analysis prepared by putting a drop of the P-dom-AgNPs suspension on a carbon-coated copper grid (300 meshes) and drying it for 30 min at room temperature (25 ◦C). A dynamic light scattering instrument (Nanotrac Wave, Microtrac, USA) was employed to estimate the average particles size and the index of polydispersity (PDI) for the synthesized AgNPs at room temperature. To estimate an average diameter of as-synthesized samples directly from FE-SEM images, Digimizer image analyzer was conducted to calculate the diameter of the particles at above 40 points. The evaluated diameters were represented as ‘‘average Feret diameter ± standard deviation”. A particle size assessment along

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a particular direction is known as the Feret/Feret’s diameter. Typically, this can be outlined as the distance between two tangential parallel lines that perpendicularly restrict the particle in that direction. This method is employed to calculate particle sizes in microscopy, where a three-dimensional (3D) particle is shown on a 2D plane [33].

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3. Results and discussion

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3.1. UV–vis absorption analysis

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The size of NPs and their quantitative development can be evaluated and recorded applying UV–vis absorption spectroscopy. In this study, UV–vis spectroscopy was employed to monitor the chemical and environmental benign syntheses of the silver particles using citrate and P-dom extract solutions. The spectral UV–vis absorption data of the synthesized Ag particles with solutions of the sodium citrate 1% w/v and P-dom extract at various pH values in AgNO3 solution (1 mM) for incubation time of 20 min at regulated temperatures (25, 45, 65 and 85 °C) are shown at Figs. 1–3, respectively. Where labels (a, b, c and d) represent the recorded UV–vis spectra from the AgNO3-P-dom extract reaction colloidal suspension as a function of extract solution concentrations (i.e. 1, 2.5, 5 and 7.5 %w/v, respectively) at the regulated temperature and pH. Through the current study, various factors including pH values (3, 5, 7 and 10), concentrations of P-dom extract (1, 2.5, 5 and 7.5 %w/v), and solutions temperature (25, 45, 65 and 85 °C) were optimized, which had been determined as parameters affecting the morphology, size and yields of AgNPs. In the current work, after adding the solution of reducing agents in AgNO3 solution, a color change from yellowish to colloidal brown was observed that visually confirmed Ag+ ions reduction to Ag° and AgNPs formation. The radiation absorption by AgNPs usually takes place in the electromagnetic spectrum visible range of 380–450 nm, due to the surface plasmon resonance (SPR) transition corresponding to the yellowish- colloidal brownish colors. This is confirmed in our study, where P-dom extract and citrate solutions did not represent any remarkable absorptions in the visible region, while bio- and chemo-synthesized AgNPs indicated intensive and distinct absorption peaks in the range of 380–450 nm that is the AgNPs SPR presence verification. Previous studies already proposed the formation of AgNPs using plant extracts/ citrate solutions and it has been shown that AgNO3 reduction to AgNPs by appropriate reducing compounds led to a distinctive surface SPR band formation, which is indicating the reduction power of these constituents. The SPR absorption peaks in the range of 400–430 nm for AgNPs with the size of around 2– 300 nm were reported in prior studies [21,29,34]. Furthermore, various investigations have revealed that silver clusters show distinct optical spectra in accordance with the number of silver atoms in that cluster. Ag° formed during the reduction of Ag ions may encounter some transformation across cluster formation. For example, the corresponding optical band to Ag2+ 8 in the absorption range varies from the typical bands for Ag2+ (310 nm) dimers, silver (360 nm), and disappears by cluster transformations of Ag2+ 8 in a silver sol [18,35–37]. Considering Fig. 1a (C = 7.5 %w/v) it is obvious that the SPR band of AgNPs centered at 450 ± 1.0 nm (pH = 10), 435 ± 1.0 nm (pH = 7), 425 ± 3.0 nm (pH = 5), and 420 ± 2.0 nm (pH = 3). The values of the SPR band of AgNPs in different pH conditions are illustrated in Fig. 3. The same trend was observed in other samples prepared at T = 65 °C (Fig. 1e–h), T = 45 °C (Fig. 2a–d), and T = 25 °C (Fig. 2e and f). It is obvious from these figures that the intensity of the SPR bands enhances as the concentration of P-dom extract and pH values raise in the reaction mediums. The

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Fig. 1. UV–vis absorption spectra of the synthesized AgNPs using the solutions of Prunus domestica extract under various pH values and extract concentrations for 20 min at treatment temperatures of 85 °C (a–d); (a) C = 7.5 %w/v, (b) 5 %w/v, (c) 2.5 %w/v, (d) 1 %w/v, and 65 °C (e–h); (e) C = 7.5 %w/v, (f) 5 %w/v, (g) 2.5 %w/v, (h) 1 %w/v.

Fig. 2. UV–vis absorption spectra of the synthesized AgNPs using the solutions of Prunus domestica extract under various pH values and extract concentrations for 20 min at treatment temperatures of 45 °C (a–d); (a) C = 7.5 %w/v, (b) 5 %w/v, (c) 2.5 %w/v, (d) 1 %w/v, and 25 °C (e–h); (e) C = 7.5 %w/v, (f) 5 %w/v. 325 326 327

noted wide SPR peaks can be originated from the broad size distribution of as-synthesized AgNPs. Wider SPR peaks at the reactions start stage, represent the large size distribution in the reactions

primary stages that become narrower as the reactions proceed. Additionally, as mentioned, the intensity enhancement in the spectrum is because of the continual Ag+ ions reduction to Ag° and

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Fig. 3. The graphs of pH values effect on the surface plasmon resonance band wavelength of the as-synthesized AgNPs using two different reducing agents (Prunus domestica extract and sodium citrate (1% w/v)); (a) T = 85 °C, (b) T = 65 °C, (c) T = 45 °C, and (d) T = 25 °C.

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increasing in AgNPs yield. Sometimes, absorption values reduction and their shifting to wavelengths with higher numbers are an evidence of aggregation as the synthesis procedures continue. It was found that augmenting the extract concentration leads to a rise in the deposited Ag yield up to 7.5 %w/v (Figs. 1 and 2). Slight shifts in the characteristic SPR peaks of this study, were implying that the primary size of the colloidal AgNPs was not impressed roughly by the enhancement in extract concentration. These results may be clarified by the fact that the carboxylate (COOA) groups of P-dom at higher pH were reduced the Ag+ ions more efficient than the lower pH, leading to AgNPs augmentation in the aqueous solution (Fig. 4). Another important affecting parameter is the temperature in metal NPs synthesis that investigated in this study. Our results showed that the reduction of Ag+ ions to Ag° could be performed at a low temperature (25 °C) in the green approach employing Pdom extract as a reducing agent. It was found that a higher temperature (80 °C) could enhance the ability of P-dom extract as reducing compounds in the reduction of Ag+ ions and formation of AgNPs. The NPs formation rate was reported to be corresponded with the reaction medium temperature, such that an enhanced temperature permitted particles to grow at a quick rate [16,22,30]. As the other influencing parameter i.e., enhancing pH values of the reaction solutions, augmented the colloidal suspension absorbance revealing higher yield of Ag-NPs (Fig. 3). Another crucial factor in the synthesis of metal NPs is the concentration of the metal precursor and its ratio to the reducing compound. We used various concentrations (ratios) of P-dom extract, while the metal precursor concentration was constant. The SPR

band blue shift, as well as the oscillator strength augmentation, are representative of a reduction in particle size because of the quantum confinement influence [38] that is evidence in Figs. 1 and 2. This can be assigned to the extract molecules inadequacy to cap the extra developed Ag nanoclusters. This shows the significance of the quantity optimizing of both AgNO3 and extract solutions added in a particular pH. In the current study, Ag+ ion was reduced to Ag° by some yet-to-be recognized biomolecules in Pdom extract. Our results revealed that at ambient temperature only solutions containing 7.5%w/v and 5%w/v of P-dom extract yield to AgNPs formation in the given reaction time (20 min) (Fig. 2e and f). While enhancing the reaction mediums temperature led to AgNPs formation at all the investigated extract concentrations. When the extract (P-dom) concentration is low, it cannot provide sufficient capping agents for nucleation of seeds. However, by increasing its concentration, the yield of the synthesized silver NPs enhance, but up to a limit. Further evaluations are needed to discover this limit via the design of experiment (DoE) to optimize the reaction. Herein, the effect of using the solution of sodium citrate 1% as a chemo-synthesizing method also investigated on the growth of AgNPs. As represented in Fig. 3a, the SPR absorbance wavelengths in UV–Vis spectra of these samples raised clearly by increasing the pH values from 3 to 10 with a red-shift at the position of the peaks. The diameter enlargement, increase in agglomeration and polydispersity of the synthesized NPs will red-shift this band to longer wavelengths, while the diameter reduction of the synthesized NPs will blue-shift this band to lower wavelengths (Fig. 3). These data were in good conformity with the previous studies

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Fig. 4. FT-IR spectra of Prunus domestica extract before and after treatment under various pH values at 85 °C.

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[21,29,36,39]. Clearly, the bio-synthesized and chemo-synthesized AgNPs, formation procedure and particle size were strongly depended on the reaction temperatures, extract concentrations, and pH values of the synthesizing solutions. Consequently, pH 10 at 85 °C was found to be the optimum value at both bio- (C = 7.5 %w/v) and chemo-synthesizing methods for achieving AgNPs with a quite uniform particles size in spherical morphology, which was additionally confirmed via the microscopic and dynamic light scattering assessments (Figs. 6–10).

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3.2. FTIR data analysis

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FTIR spectroscopy is a valuable method to study existence probabilities of various functional groups from the peak position in the IR spectrum. Fig. 4 shows the FTIR spectrum of the pure extract of P-dom and the samples prepared using 7.5 %w/v extract solution at 85 °C in pH values of 3–10 for 20 min. In this work, to discover which functional groups were concerned with Ag+ ions reduction to Ag°, the FTIR data prior and later the reduction were compared. Several functional groups of P-dom extract before bioreduction were detected at approximately 691, 1037, 1373, 1417, and 1638 cm
1 originated from various present biomolecules in the aqueous extract of P-dom. These spectral bands could be ascribed to the NAH wag of amines, amines CAN stretching/CAO stretching vibration, OAH bending, CAH bond bending vibration, (NH)C@O group stretching vibration, respectively. Furthermore, the other spectral bands at 2250, 2840, 2891, 3300 and 3450 cm
1 could be ascribed to glycosidessy CAN triple bonds (nitriles groups) stretching, CAH stretching merged with a broad carboxylic acid peak, aldehydes CAH stretching vibrations, a broad OAH stretching band (including hemiacetal, ACOOH, and ether groups), and polyols OAH stretching vibrations, respectively. The secondary metabolites existence like terpenoids, amino acids, glycosides,

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Fig. 5. (a) XRD patterns of synthesized Ag nanoparticles using Prunus domestica extract and citrate solutions under various pH values at 85 °C; (b) typical EDX pattern of AgNPs synthesized by P-dom extract solution at pH 5.

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Fig. 6. FE-SEM images of the as-synthesized Ag nanoparticles at 85 °C by Prunus domestica extract under controlled pH = 3 and various extract concentrations (a) 1 %w/v, (b) 2.5 %w/v, (c) 5 %w/v, and (d) 7.5 %w/v; (a0 ), (b0 ), (c0 ) and (d0 ) show the correspondent size distribution histograms (DLS data) of the represented images, respectively.

Fig. 7. FE-SEM images of the as-synthesized Ag nanoparticles at 85 °C by Prunus domestica extract under controlled pH = 5 and various extract concentrations (a) 1 %w/v, (b) 2.5 %w/v, (c) 5 %w/v, and (d) 7.5 %w/v; (a0 ), (b0 ), (c0 ) and (d0 ) show the correspondent size distribution histograms (DLS data) of the represented images, respectively.

Fig. 8. FE-SEM images of the as-synthesized Ag nanoparticles at 85 °C by Prunus domestica extract under controlled pH = 7 and various extract concentrations (a) 1 %w/v, (b) 2.5 %w/v, (c) 5 %w/v, and (d) 7.5 %w/v; (a0 ), (b0 ), (c0 ) and (d0 ) show the correspondent size distribution histograms (DLS data) of the represented images, respectively.

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Fig. 9. FE-SEM images of the as-synthesized Ag nanoparticles at 85 °C by Prunus domestica extract under controlled pH = 10 and various extract concentrations (a) 1 %w/v, (b) 2.5 %w/v, (c) 5 %w/v, and (d) 7.5 %w/v; (a0 ), (b0 ), (c0 ) and (d0 ) show the correspondent size distribution histograms (DLS data) of the represented images, respectively.

Fig. 10. FE-SEM images of the as-synthesized Ag nanoparticles at 85 °C by citrate 1%w/v under controlled pH values (a) 3, (b) 5, (c) 7, and (d) 10; (a0 ), (b0 ), (c0 ) and (d0 ) show the correspondent size distribution histograms (DLS data) of the represented images, respectively.

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phenols, and flavanols was reported in P-dom fruit extract [14,29,35]. Taking into account the FTIR data, make it feasible to realize not only the possibility of surrounding synthesized NPs by some proteins or metabolites (such as carboxylate, amine, alcohol, aldehyde, ketone, or other functional compounds), but also NPs stabilization caused by functional groups of the reducing agents. Mostly, fractions of the green reducing agents that are soluble in water play an intricate role in bioreduction of metal ions and related morphology development of the metal NPs that can be evaluated by FTIR. Furthermore, this can be conducted to investigate the role of the bioreductants (capping ligand and reducing agent) in metal NPs stabilization in the solvent [40]. Considering the FTIR spectra of the samples after reduction Ag ions using P-dom extract showed that biosynthesized samples have the alkanes stretching vibrations of ACH groups, polyphenolic AOH and stretching of the carboxylic acid group with slight reduction in intensity and shifts. This observation signifies the involvement of functional groups of P-dom extract biomolecules in Ag ions reduction to the corresponding AgNPs. Similar results have been published for biofabrication of Au ions, declaring these func-

tional groups involved in Au ions reduction to AuNPs using appropriate extracts [14,41]. The observed minor shifts of the bands may be related to the Ag ions interaction with the functional groups of P-dom extract. Furthermore, appearing the characteristic FTIR peaks of P-dom in the samples treated at 85 °C with various pH values (pH 3–10) in approximately identical to the original P-dom peaks revealed that no significant alterations occurred in the principal FTIR peaks of P-dom after treatment. These results suggest that the main chemical structure of P-dom was practically preserved. In the current study, bioreduction reactions were developed in an aqueous medium. Since biomolecules with low aqueous solubility (terpenoids) were not the main involved moieties in the bioreduction procedure. It was deduced that bio-organics of P-dom extract, for instance, various enzymes using free amino groups of proteins bind to the Ag atoms and are corresponding for NPs synthesis and their stabilization. Amino groups functioned as a thin film on molecules and averted NPs agglomeration. In other words, the suggested mechanism for synthesis of AgNPs is attributed to the hydroxyl groups, flavonoids, and alkaloids. Where hydroxyl groups form hydrated electrons and they are responsible for Ag+

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ions reduction to Ag0 [42]. Furthermore, it has mentioned that the Ag+ ions are caught on the alkaloids surface and are subsequently reduced via the other biomolecules resulting in the Ag nuclei materialization. These produced Ag nuclei accumulate and then grow in size leading to the formation of AgNPs [35]. The polyhydroxy group, with flavonoids as a secondary metabolite are the other main reducing agents of aqueous plant extracts participating in the metal ions reduction into the corresponding nanoparticles [43]. The active hydroxyl groups existing in flavonoids have a crucial role in their antioxidant characteristics with free radicals scavenging or the metal ions chelating [34,43]. In another word, the reducing or antioxidant behaviors of flavonoids are linked to their hydrogen atoms or electron donation ability. Consequently, the AgNPs formation employing these compounds of P-dom might be because of Ag+ ions chelation that directed to easy electron transmit from the oxidant state to the reductant state [39]. The overall reaction of Ag+ ions reduction to Ag0 in chemosynthesizing method while sodium citrate 1% solution is blended with the solution of silver nitrate at a high temperature can be outlined by the following equation:

482 2
þ þ 0 C 6 H5 O3
7 þ 2Ag ! C 5 H 4 O5 þ H þ CO2 þ 2Ag

484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510



ðC 5 H4 O2
5 ¼ OOCCH2 COCH2 COO Þ

Citrate plays numerous roles in the formation of AgNPs. At the ambient temperature, it forms an Ag+-citrate complex (pK1 = 7.1) by firmly coordinating with Ag+ ions. At an elevated temperature acting as a reducing agent, it generates Ag metal particles. Citrate also acts as a capping compound to preserve the lately formed Ag nuclei. The kinetic characteristics of the chemo-synthesized AgNPs and their homogeneous nucleation have been explained in literature [36,44,45]. Briefly, the colloidal particles formation in a solution under homogeneous nucleation occurs in a series of steps, which can be clarified as the following steps: (I) Nucleation process, namely the formation of Ag0 atoms with the creation of nuclei i.e., nAg þ ! nAg 0 ! Ag 0n And based on the interrupted (step by step) mechanism 2þ 0 i.e., Ag þ ! Ag 0 ! Ag þ 2 ! Ag 4 ! :::: ! Ag n The rate of this nucleation stage will be controlled by chemically reduction rate of Ag ions. (II) (Growth stage) following the new phase nucleus formation, NPs growth has begun This can happen both in correspondence with a coagulative mechanism i.e., mAg 0n ! Ag 0n:m And Ag ions reduction on the nucleus surface i.e., Ag 0n þ Ag þ þ e
! Ag 0nþ1 Definitely, in actual systems, all these procedures will occur simultaneously and complete each other.

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3.3. X-ray diffraction and energy-dispersive X-ray studies

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The XRD measurements were performed through the current study to evaluate the crystalline structure and phase configuration of the bio/chemo-synthesized AgNPs. Fig. 5a illustrates the representative XRD patterns of AgNPs prepared by P-dom extract solution at pH 3–10 and citrate 1%. All the main peaks in Fig. 5a can be classified as face-centered cubic (fcc) structure of Ag crystals with a lattice constant of a = 4.08 Å, which are in good agreement with the database of the Joint Committee on Powder Diffraction Standard (JCPDS No. 04-0783 and No. 03-0921). The four typical peaks at 2h angles corresponding to 38.30◦, 46.60◦, 64.80◦, and 77.50◦ can be ascribed to diffraction from the planes of (1 1 1), (2 0 0), (2 2 0), and (3 1 1), respectively, caused by the Bragg’s

513 514 515 516 517 518 519 520 521 522 523

9

reflections of the fcc crystal structure of Ag. These are in good conformity with previous studies [21,34,46]. Furthermore, some minor peaks revealing the hexagonal crystal structure (JCPD No. 41–1402) at 2h angles corresponding to 44.16°, 57.46°, 67.36° can be ascribed to diffraction from the planes of (1 0 0), (1 0 3), and (0 0 6), respectively. It implies a biphasic nature of the synthesized AgNPs. Three yet unassigned peaks at 2h angles corresponding to 28.2◦, 32.6° and 57.8° in XRD pattern of the prepared AgNPs appeared which may be bound to the biological agents in P-dom extract as a stabilizing and capping compound. It suggests that the bioorganic phase crystallization takes place on the surface of AgNPs [21,34,46]. It should be noted that extremely sharp diffraction peaks could be attained from a perfect sample, where usually in practice diffraction patterns of powders show broad peaks that are evaluated in degrees hundredths. The peaks broadening is due to the distortion of the crystal lattice (microstrain caused by concentration gradients and dislocations), diffracting domain /crystallite size alterations, structural defects (i.e., stacking faults, twin faults) and etc. Fig. 5b illustrates the typical EDX spectrum of AgNPs synthesized by P-dom extract solution at pH 5, along with its elements weight percentage. It is obvious that a detected strong characteristic peak at approximately 3.0 keV is the silver distinctive absorption signal corresponding to AgNPs SPR. Furthermore, some peaks of O were also detected, possibly triggered by P-dom extract involvement in the AgNPs formation. The EDX spectrum evaluation also demonstrated that the AgNPs are in a metallic state with no Ag2O formation in them and free from any other impurities. XRD and EDX data are in good agreement with UV–vis data (Figs. 1 and 2), the morphological and particles size distributions assessments (Figs. 6–11) as well as with the previous studies that had been revealed similar results [21,29,34,43,46].

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3.4. Morphological and particles size distributions assessments

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It is vital to identify AgNPs size and morphology following production confirmation using P-dom extract and citrate solutions through the reactions to comprehend how various reducing compounds and reaction environments can influence the shape, size and synthesized AgNPs yield. In this regard, FE-SEM, TEM, and DLS analyses were conducted to evaluate morphologies as well as particles size and polydispersity index (PDI) of the resulting samples. Fig. 6 shows the obtained data of AgNPs evaluation prepared using various extract concentrations (a) 1 %w/v, (b) 2.5 % w/v, (c) 5 %w/v, and (d) 7.5 %w/v at 85 °C in pH value of 3. By considering Fig. 6a–d and Fig. 6a0 –d0 , it was found that the average particle sizes of AgNPs were 16.33 ± 5, 32.16 ± 4.8, 44.28 ± 3, and 46.86 ± 2.9 nm corresponded to different concentrations of P-dom extract (Fig. 6a–d, respectively). Where, the average particles size derived from DLS analysis and the PDIs of the biosynthesized AgNPs were found to be 84.9, 4.78 at C = 1%w/v; 118.4, 4.12 (2.5%w/v), 133.4, 1.31 (5%w/v), and 197.5, 1.08 (7.5% w/v), (Fig. 6a0 –d0 , respectively). Table 1 summarize the calculated average particles size from FE-SEM images (Feret diameter) of AgNPs as well as their mean hydrodynamic diameter and PDIs for Fig. 7 (pH = 5, biosynthesized), Fig. 8 (pH = 7, bio-synthesized), Fig. 9 (pH = 10, biosynthesized), and Fig. 10 (chemo-synthesized, citrate 1%). Considering Table 1, and Figs. 6–10 reveal the effect of various extract concentrations and reducing agents on morphology and particles size of AgNPs. It is obvious from Figs. 6–10 and Table 1 that, while the pH value of P-dom extract solution was decreased from 10 to 3, the prepared AgNPs were increased in the particles size. Furthermore, these data illustrate an enhancement in particle size of bio-

557

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Fig. 11. TEM images of the as-synthesized Ag nanoparticles by Prunus domestica extract under controlled pH values at 85 °C; (a) pH 3, (b) pH 5, (c) pH 7, and (d) pH 10.

Table 1 Feret diameter ± standard deviation (FE-SEM), mean hydrodynamic diameters (DLS) and polydispersity index (PDI) of bio- and chemo-synthesized AgNPs (based on Figs. 7–10).

Biosynthesized AgNPs

Chemosynthesized AgNPs

587 588 589 590 591 592 593 594 595 596 597 598 599 600 601 602 603 604

pH 5

%w/v

pH 7

%w/v

pH 10

%w/v

pH

3 5 7 10

synthesized AgNPs with augmenting concentrations of P-dom extract. As it can be inferred from the Table 1, changes in reducing agents altered AgNPs size proposing that P-dom and citrate must act as a nucleation driving/controller and stabilizer. In all the samples reduced with the high concentration (7.5 %w/v) of P-dom extract solution (Figs. 6d, 7d, 8d, and 9 d) anisotropic particles with a relatively large diameter (around 50 nm) were formed. While smaller ones were achieved at the low concentration of P-dom extract with an average diameter of about 20 nm. Clearly, these data revealed the remarkable particle size and morphology dependence of the as-synthesized AgNPs on the pH values and concentrations of P-dom extract and citrate solutions. This has been reported by other researchers that adjusting pH values of reducing solutions and their concentrations could affect the size and shape of the synthesized AgNPs [46,47]. The differences in the measured particles values raised from the mechanism of DLS technique that calculate a hydrodynamic diameter of particles.

1 2.5 5 7.5 1 2.5 5 7.5 1 2.5 5 7.5

FE-SEM

DLS

PDI

21.43 ± 4.00 35.67 ± 2.10 46.81 ± 3.00 50.31 ± 5.10 19.87 ± 3.00 35.55 ± 4.50 38.34 ± 2.85 51.43 ± 2.34 20.08 ± 2.15 39.21 ± 4.01 46.47 ± 2.14 58.17 ± 4.00 6.68 ± 1.12 24.32 ± 3.28 43.97 ± 5.01 59.95 ± 2.69

38.00 84.90 196.80 184.00 30.50 57.40 87.60 179.00 15.52 42.10 48.40 57.20 3.82 22.69 29.18 50.00

4.89 4.78 1.14 5.52 2.91 8.36 4.10 1.29 5.03 2.45 4.24 5.24 3.34 2.79 1.19 3.35

The synthesized AgNPs through employing P-dom extract (7.5% w/v at 85 °C) with the highest UV–vis peaks during the reaction was further analyzed by TEM technique. TEM assessments of biosynthesized samples using P-dom extract (7.5 %w/v) at 85 °C with varying pH values (a) 3, (b) 5, (c) 7, and (d) 10 are depicted in Fig. 11. TEM observations indicated that the bio-synthesized AgNPs morphology at pH = 3 was mainly polygonal with an average particles size of 16 nm (Fig. 11 a). The samples prepared at pH = 5 represented oval morphologies with an average particle size of 30 nm (Fig. 11 b), the synthesized AgNPs at pH = 7 showed semi-spherical morphologies with a mean diameter of 40 nm (Fig. 11c), and AgNPs synthesized at pH = 10 indicated spherical morphologies with an average diameter of 50 nm (Fig. 11d). As witnessed the effects of various pH values and the extract solution concentrations of P-dom on the UV–vis absorption spectra (Figs. 1–3) and the XRD results (Fig. 5), different sizes of assynthesized AgNPs were profoundly confirmed by FE-SEM, TEM and DLS observations (Figs. 6–11). While the pH value of P-dom

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extract solution was increased from 3 to 10, the prepared AgNPs were noticed to acquire an approximately spherical morphology with an increase in the particles size. These findings proposed that the pH values have tremendous effect on the morphology and size of the synthesized AgNPs that might be related to the chain conformation alterations of P-dom extract under different pH conditions (Fig. 4). As previously mentioned, a red shift in SPR peaks occurs when the particles diameter increase, as it is clear from Fig. 3 the SPR peaks of the bio-synthesized samples revealing a particles size distribution as witnessed by TEM and DLS data (Figs. 6 and 11). Furthermore, no variation in visual aggregation and colors has been witnessed even after storage for one month (data not represented). Consequently, the bio-synthesized AgNPs at various pH values (5–10) are well distributed in nature and highly stable. Through the current study, it was revealed that the pH 10 produce AgNPs with spherical morphology and relatively uniform size distribution with larger average particle size.

[7]

[8]

[9]

[10]

[11] [12] [13] [14]

[15] 640

4. Conclusion

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Comparative facile and inexpensive chemically as well as environmental benign AgNPs synthesizing procedures employing citrate and prunus domestica compounds under aqueous environments were outlined through the current work. The experimental setups under various conditions were provided to control the size and morphology of the synthesized AgNPs. Additionally, feasible functional groups and mechanisms ruling a stimuli-responsive AgNPs formation using citrate 1% and P-dom extract solutions were given. FTIR analysis proposed that the aqueous-soluble polyols such as glycosides, phenols, and flavanols were corresponded for Ag ions reduction to AgNPs. Bio-synthesized AgNPs indicated nearly spherical morphology and crystalline structure with average particle sizes of 20–50 nm (FE-SEM and TEM). The investigated colloidal AgNPs using DLS method showed a slightly higher average diameters. The as-synthesized AgNPs by both chemo- and bio-synthesizing methods had a quite well-dispersed spherical morphology and larger particle sizes under alkali conditions (pH 10) based on UV–vis spectroscopy, XRD graphs, FE-SEM/TEM observation, and DLS data. Our study demonstrates that the reaction mixtures initial pH and concentration of the used extract solution affects the SPR, stability, morphology, and size of colloidal AgNPs. The ability to modify the size and morphology of nanoparticles as presented in the current study for AgNPs opens up an interesting opportunity of further synthetic processes employing biological sources.

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Acknowledgments

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[27]

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The was study supported by Elite Researcher Grant Committee under award number (No. 963285) from the National Institutes for Medical Research Development (NIMAD), Tehran, Iran.

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