Green Chemistry Letters and Reviews

ISSN: 1751-8253 (Print) 1751-7192 (Online) Journal homepage: https://www.tandfonline.com/loi/tgcl20

Biosynthesis of silver nanoparticles with adiantum

capillus-veneris L leaf extract in the batch process
and assessment of antibacterial activity

Sariyeh Omidi, Sajjad Sedaghat, Kambiz Tahvildari, Pirouz Derakhshi &

Fereshte Motiee

To cite this article: Sariyeh Omidi, Sajjad Sedaghat, Kambiz Tahvildari, Pirouz Derakhshi &
Fereshte Motiee (2018) Biosynthesis of silver nanoparticles with adiantum�capillus-veneris�L leaf
extract in the batch process and assessment of antibacterial activity, Green Chemistry Letters and
Reviews, 11:4, 544-551, DOI: 10.1080/17518253.2018.1546410

To link to this article: https://doi.org/10.1080/17518253.2018.1546410

© 2018 The Author(s). Published by Informa Published online: 19 Nov 2018.

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GREEN CHEMISTRY LETTERS AND REVIEWS
2018, VOL. 11, NO. 4, 544–551
https://doi.org/10.1080/17518253.2018.1546410

Biosynthesis of silver nanoparticles with adiantum capillus-veneris L leaf extract in

the batch process and assessment of antibacterial activity
Sariyeh Omidia, Sajjad Sedaghatb, Kambiz Tahvildaria, Pirouz Derakhshia and Fereshte Motieea
a
Department of Chemistry, College of Science, North Tehran Branch, Islamic Azad University, Tehran, Iran; bDepartment of Chemistry, College of
Science, Shahr-e-Qods Branch, Islamic Azad University, Shahr-e-Qods, Iran

ABSTRACT ARTICLE HISTORY

The aqueous extract of Adiantum capillus-veneris L was used as a reducing agent in the batch Received 15 February 2018
method for the synthesis of silver nanoparticles (AgNPs). For improving AgNP synthesis, the Accepted 6 November 2018
concentrations of metal ions solution and leaf extract were optimized in 24 h. Using Fourier
KEYWORDS
transform infrared spectroscopy (FTIR), powder X-ray diffraction (XRD), transmission electron Biosynthesis; silver
microscopy (TEM), and UV-Vis spectroscopy, AgNPs were characterized. In the UV-Vis spectrum, nanoparticles; Adiantum
the peak was observed at 453 nm, indicating the surface plasmon resonance (SPR) of AgNPs. The capillus-veneris L;
purity and crystalline nature of AgNPs were confirmed based on the XRD assessment. In antibacterial activity; batch
addition, specific functional groups, which led to silver nitrate reduction in AgNP formation, were reactor
examined using FTIR. According to TEM, the average particle size was 18.4 nm in AgNPs.
Antibacterial activity assays with Escherichia coli and Staphylococcus aureus demonstrated that
AgNPs reduced bacterial growth and produced well-defined inhibition zones.

Introduction
Nanotechnology development and research have grown
rapidly around the world (1,2). Today, AgNPs are recog-
nized as commonly used NPs with a variety of applications
(3). They are used as optical receptors, coating agents in
solar energy absorption, and biological labels (4).
For the synthesis of metal NPs, different metals includ-
ing Ag, Pd, Zn, Au, and CdS have been applied (5–7).
Among metal NPs, AgNPs show significant physical,
chemical, and biological properties (8), with potential
antimicrobial and anticancer applications in different
industries (e.g. textile, food packaging, and cosmetic
industries) (9–12). Various chemical, physical, and bio-
logical approaches have been examined for the synthesis
of AgNPs (13). Although chemical synthesis is generally a
rapid process, it necessitates the use of toxic chemicals,
such as NaBH4, N2H4, NH2OH, and ethanol (3,14).
Moreover, control of the growth, stability, and aggrega-
tion of particles is difficult through chemical synthesis,
and for stabilizing the NP size, use of capping agents is
necessary (3). In recent years, use of plant extracts has
been highlighted for NP synthesis. Considering its non-
toxicity, simplicity, and large-scale production, appli-
cation of biosynthetic green metal NPs is growing
(1,15–19). Various parts of plants, including the stem,
root, leaf, flowers, and fruits, contain phytochemical sub-
stances and function as stabilizing and reducing agents
during NP formation (20). Recently, extracts of various
plant such as Nigella sativa (3), Tragia involucrata (4),
Ulva flexuosa (9), Achillea biebersteinii (15), Lonicera japo-
nica (16), pennyroyal (21), olive (22), Acalypha indica (23),
Azadirachta indica (24), Morinda pubescens (25), tea (26),
and Rosmarinus officinalis have been reported to aid in
the formation of NPs ranging in size from 10 to 33 nm

CONTACT Sajjad Sedaghat Sajjadsedaghat@yahoo.com

© 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.

(27), and have been further explored as a source for the
synthesis of AgNPs. Also previous studies have been
reported synthesis of silver nanoparticles via Adiantum
capillus-veneris L (28). Active compounds found in Adian-
tum capillus-veneris L include flavonoids, terpenoids, phe-
nolic acids, alkaloids, fiber (29,30). In our rapid and simple
method, Adiantum capillus-veneris L leaf extract was used
for the synthesis of AgNPs as reducing and capping
agents under normal atmospheric conditions in the
batch reactor. This is a report on Adiantum capillus-
veneris L-assisted synthesis of AgNPs in large-scale, pre-
pared using low amounts of leaf extracts, without requir-
ing any extra compounds or physical processes.
In many countries, the significant biological activity,
extensive availability, and easy collection of Equisetum
arvense have led to its medicinal use. Recently, various
antimicrobial activities have been attributed to Adiantum
capillus-veneris L (31). Use of the extracts of Adiantum
capillus-veneris L does not require any intermediate
base groups and is considered cost-effective. In addition,
it is not time-consuming, does not require expensive pre-
cursors or equipment, and yields a pure product without
impurities.
Adiantum capillus-veneris L is recognized as the best
source of NP synthesis, as it leads to large-scale pro-
duction of stable NPs in different sizes and shapes. The
leaf extract and metal solution concentrations were opti-
mized to improve AgNP synthesis. AgNPs were charac-
terized using several types of spectral analysis. For
evaluating the antibacterial activity of silver NPs, the
disc diffusion method was applied.
Materials and methods
Preparation of Adiantum capillus-veneris L
aqueous extract
Green Adiantum capillus-veneris L leaves were collected
from North of Iran in June 2016. After washing the
leaves, they were dried under indirect light for 7 days
and ground into powder; the powder was stored at a
temperature of 25°C. Merck Co. provided AgNO3
(99.80%). By adding double-distilled water, the aqueous
solution was prepared. Adiantum capillus-veneris L leaf
powder (50 g) was incubated for 15 min in a 70°C
water bath. Afterwards, the extract was centrifuged for
15 min at 4000 rpm to eliminate heavy biomaterials.
Synthesis of silver nanoparticles
The Adiantum capillus-veneris L aqueous extract was
mixed with 1, 2, or 3 mM AgNO3 (1000 mL), at 25°C for
24 h via mechanical Stirring at 750 rpm. At room temp-
erature, the color changed from yellow to dark brown.
GREEN CHEMISTRY LETTERS AND REVIEWS 545
Purification of silver nanoparticles
AgNPs were collected gradually during the reaction. For
the removal of silver colloid residues, centrifugation was
carried out for 15 min using double-distilled water at
4000 rpm; then, the solution was rinsed with deionized
water three times. For further characterization, the
dried AgNP powder was collected following incubation
for three hours at 65°C (32,33).
Ultraviolet–visible (UV–Vis) absorption
spectroscopy
Formation of AgNPs was monitored at 300–800 nm,
based on the UV-Vis spectra of the reaction medium.
For recording the spectra, a UV-Vis spectrophotometer
(BioTek) was employed. Before every measurement, the
samples were diluted in water.
Fourier transform infrared (FTIR) spectroscopy
Spectroscopy was performed to determine the func-
tional groups in AgNP synthesis, which were bound dis-
tinctively to the AgNP surface. Before and after AgNP
synthesis, the Adiantum capillus-veneris L extract was
vacuum-dried to prepare the samples. Afterwards, the
dried samples were mixed with potassium bromide
and pressed into a sheer slice. An FTIR spectrometer
(Perkin Elmer, USA) was used to examine the samples.
X-ray diffraction (XRD)
The crystal structure of AgNPs was determined and
confirmed via XRD analysis. After placing the air-dried
NPs on an XRD grid, they were assessed in terms of
AgNP formation, using an X-ray diffractometer (Cu Kα
radiation, θ–2θ configuration; PANalytical) with an
X’Pert Pro generator (30 mA; 40 kV). For measuring the
crystallite domain size, the XRD peak width was deter-
mined under the assumption that non-uniform strains
are absent. The Scherrer equation was calculated as
follows (Equation 1):
0.9l
D=
bcosu
where D denotes the average crystallite domain size per-
pendicular to the reflecting planes, λ presents the X-ray
wavelength, β represents the full width at half
maximum (FWHM), and θ indicates the diffraction angle.
Transmission electron microscopy (TEM)
A Zeiss TEM microscope (EM 10C/CR, Zeiss, Germany)
was used for the morphological assessment of AgNPs
546 S. OMIDI ET AL.

at 100 kV. After drop-casting the suspension on a carbon-

coated copper grid, it was left to dry overnight at 25°C.

Antibacterial activity
For determining the antibacterial activity of AgNPs, agar
disk diffusion was used according to the literature (34–
37). Briefly, using broth dilution, AgNPs at 5, 0.5, and
0.05 μg/mL were used to measure the minimum bacteri-
cidal concentration for preventing Escherichia coli (ATCC
25922) and Staphylococcus aureus (ATCC 25923) growth.
Bacterial cultures at a concentration of 1.5 × 108 CFU/mL
were inoculated with different concentrations of AgNPs.
After incubating the culture plates at 37°C for 24 h, the
bacterial growth inhibition zone was measured in centi-
meters. For each bacterial strain, three independent
experiments were performed (38,39).

Results and discussion

To investigate whether an aqueous extract of Adiantum
capillus-veneris L can be used in the green synthesis of
AgNPs, we combined the extract with 1, 2 and 3 mM of
AgNO3 for 24 h. The slow change of color in the solution Figure 1. UV visible absorption spectrum showing (a) Optimum
(from yellow to dark brown) was indicative of AgNP for- AgNO3 concentration, (b) Optimum concentration of Adiantum
mation. The dark brown color might be related to the capillus-veneris L extract.
surface plasmon resonance (SPR) effect and AgNO3
reduction (40).

UV–VIS spectroscopy
UV–Vis spectroscopy is described as a suitable tool for NP
assessment, especially in stability analysis of metal NPs in
aqueous solutions. To determine if the reaction had
reached completion, different concentration of AgNO3
(1, 2 and 3 mM) was reacted with different concentration
of the aqueous extract (5, 10 and 15 mL), and the spectra
were recorded at a range of 300–800 nm (41); then, the
wavelength of maximum absorption was identified Figure 2. UV–VIS spectra of synthesized AgNPs with using opti-
(Figure 1). Considering the color changes after the mized AgNO3 and Adiantum capillus-veneris L extract.
addition of AgNO3, AgNP biosynthesis by the Adiantum
capillus-veneris L extract was confirmed.
XRD analysis
According to the analysis, a sharp SPR related to 3 mM
AgNO3 and 5 mL aqueous extract was observed at nearly Using the XRD pattern of synthesized AgNPs, distinctive
453 nm (Figure 2), which is in line with reports of peaks in peaks were identified, indexed to the crystalline planes
the absorption spectra of 400–500 nm associated with of silver (Figure 3); these results conform to the JCPDS
SPR in AgNPs (14,42–44). The NPs were dispersed reports (No, 04-0783). The crystalline nature of syn-
without using any chemical or physical capping agents. thesized AgNPs was confirmed considering the sharp
AgNPs synthesized with Adiantum capillus-veneris L peaks (4). Intense NP peaks of (1 1 1), (2 0 0), (2 2 0),
extract showed stability in the solution. The reported and (3 1 1) emerged, indexed as the crystalline silver
size of these synthesized AgNPs was smaller than many face-centered-cubic (FCC) phase (46). The mean particles
earlier green synthesis reports (43,45). size of AgNPs was evaluated using the Debye–Scherer

Figure 3. XRD pattern of AgNPs, indicating crystal structure.
Equation 1(Equation 1):
Kl
d=
bcosu
where λ denotes the X–ray wavelength (1.540560 Å), β
represents the width of XRD peak, θ shows the Bragg
angle, d refers to the size of particle, and K describes
the Scherer constant (0.9) (14,47). The average size of
AgNPs was almost around 25 nm.
FTIR analysis
For determining the biomolecules, resulting in AgNP
reduction and stabilization, FTIR analyses were
Figure 4. (a) FTIR spectra of Adiantum capillus-veneris L extract, (b) Silver nanoparticle synthesized using Adiantum capillus-veneris L
extract.
GREEN CHEMISTRY LETTERS AND REVIEWS 547
performed. In Figure 4, the FTIR peaks of dried Adiantum
capillus-veneris L and AgNPs are presented (4). The FTIR
results revealed absorption bands at 3396.66, 1625.75,
and 1427.55 cm−1 (Figure 4(a)). The vibrational bands
corresponded to bonds of aminos (=N−H), alcohols
(−O−H), alkenes (>C=C), flavonoids, and amines
(−NH2), all in the range of 800–3442 cm−1. Bonded
hydroxyl (−OH) groups of carboxylic acids and phenols
could produce absorption bands at 3396.66 cm−1
(Figure 4(a)). In the AgNP spectrum, the new band at
1607.23 cm−1 might suggest a new C=O group (a
ketone or an aldehyde) (Figure 4(b)).
For identifying biomolecules, which may be involved
in the capping and reduction of synthesized AgNPs,
FTIR measurements were performed. The FTIR spectra
of the extract indicated absorption bands, corresponding
to functional groups including phenols, alcohol, amines,
and carbonyls. This strongly suggests that hydroxyl
groups reduce Ag+ and are oxidized to carbonyl
groups in the process (Figure 5) (48).
AgNPs show significant binding affinity to multiple
functional groups in the compounds of plants (e.g. poly-
phenolics and proteins); a layer of these functional
groups covers AgNPs. They act as capping agents, pro-
viding stability and preventing AgNP agglomeration, as
confirmed in the green synthesis of AgNPs with other
plant extracts (49,50). These findings confirm that pheno-
lic agents in the Adiantum capillus-veneris L extract are
responsible for the reduction of AgNO3.
548 S. OMIDI ET AL.

Figure 5. Mechanism of Ag NPs formation from flavonoids and polyphenols present in the leaf extract of Adiantum capillus-veneris L.

Figure 6. (a) TEM image of size of silver nanoparticles synthesized by Adiantum capillus-veneris L extract, (b) Particle size distributions of
biosynthesized AgNPs.

TEM analysis Antibacterial activity

TEM imaging was applied to measure AgNP size and
surface morphology in the solution. As shown in
Figure 6(a), the TEM study demonstrated that the
average size was 18.4 nm with interring particle dis-
tance. The morphology of Ag nanoparticles was
also spherical (Figure 6(a)). In another work, Devaraj
(51) synthesized AgNPs from Diospyros montana
extract and obtained AgNPs in variable size and
morphology.
For evaluating the antibacterial activity of AgNPs and
aqueous Adiantum capillus-veneris L extract, both Gram-
positive (S. aureus) and Gram-negative (E. coli) bacteria
were used (Figures 7 and 8).
The maximum inhibition zone in a highly concen-
trated solution (stock AgNPs) is listed in Table 1. Also,
the zone inhibition of two antibiotics Ciprofloxacin and
Gentamicin was compared with AgNPs (52). AgNPs
were as effective as Ciprofloxacin and Gentamicin. Also,

Figure 7. Antibacterial activity of AgNPs against (a) Gram negative (E. coli), (b) Gram positive (S.aureus) in different solutions.
Figure 8. Antibacterial activity of Adiantum capillus-veneris L extract against (a) Gram negative (E. coli), (b) Gram positive (S.aureus) in
different solutions.
silver not only have a close interaction with the surface of
the membrane, but can also penetrate deep inside the
bacteria. According to some studies, the antimicrobial
activity of silver is relatively better against Gram-negative
bacteria, compared to Gram-positive bacteria; this may
be related to the thinner layer of peptidoglycans and
beta barrel proteins (porins) in the cell wall structure
(53). This mechanism is classified broadly as the direct
effects of NPs on microbial cells. The cell wall becomes
permeable to the extracellular medium as soon as the
cell membrane is damaged, and bacterial cells become
susceptible to damage (54).
Conclusion
The biosynthesis of stable AgNPs from an aqueous
AgNO3 solution was revealed, using Adiantum capil-
lus-veneris L as the reducing and capping agent. Also
this method is easily adopted mass production of
GREEN CHEMISTRY LETTERS AND REVIEWS 549
nanoparticles without requiring any extra compounds
or physical processes and have well-defined size and
morphology. Optimization of AgNO3 and Adiantum
capillus-veneris L extract concentration was achieved
to obtain large amount of silver nanoparticles in
batch process. UV–Vis, FTIR, TEM, and XRD studies
were performed to confirm the AgNP formation. For
AgNPs, UV–Vis peak was observed at 453 nm. The crys-
talline structure of synthesized AgNPs was determined
using XRD analysis. The peaks in XRD patterns were
associated with those of the FCC form of metallic
silver. AgNPs had a quasi-spherical shape and an aniso-
tropic nature (average size, 18 nm). The strong antimi-
crobial activity of biosynthesized NPs was confirmed
against Gram-positive (S. aureus), as well as Gram-nega-
tive (E. coli) bacteria. The green synthesis of AgNPs
with the extract is a more cost-effective and eco-
friendly strategy, compared to physical and chemical
synthesis methods.
550 S. OMIDI ET AL.

Acknowledgements

resistance to

resistance to
antibiotic

antibiotic
This work was supported by North Tehran branch and Shahr-e-

0.05
0

0
Qods Branch of Islamic Azad University of Iran. We thank these
two Universities for their kind Cooperation.

resistance to
Disclosure statement

sensitive to

antibiotic
0.5
5

0
AgNPs
AgNPs (μg/Ml) No potential conflict of interest was reported by the authors.
Gram negative (E.coli)

Funding

resistance to
sensitive to

antibiotic
12

0
5

This work was supported by North Tehran branch and Shahr-e-

AgNPs Qods Branch of Islamic Azad University of Iran.
Thick nano particle

Sensitive to AgNPs

Notes on contributors
suspention

resistance to

Sariyeh Omidi received M.Sc. degree in applied chemistry from

19

antibiotic
0

Islamic Azad University-Shahre Ray, Yadegare Emam Branch,

Tehran, Iran, in 2010. She is a Ph.D. Student in Applied Chem-
istry in Islamic Azad University-Tehran North Branch since
2014. Her main areas of interests are Nano technology, Green
Ciprofloxacin

chemistry and water treatment.

antibiotic

resistance to

sensitive to
antibiotic
0

28

Sajjad Sedaghat received the M.Sc. degree in organic chemistry

AgNPs

from Shahid beheshti University, Tehran, Iran, in 2002. He

received his PhD. in Applied chemistry from Islamic azad uni-
versity, north Tehran branch in nano catalyst for fuel cells in
2006. Since 2006, he has been with Islamic azad university,
Bacterial strains

Resistance to

resistance to
antibiotic

Shahr-e-Qods branch, Tehran, Iran as a lecturer in the green

0.05
0

0
AgNPs

chemistry group. His main areas interests are nano synthesis,

nanocomposites and Green chemistry.
Kambiz Tahvildari received M.Sc. degree in applied chemistry
from Islamic Azad University-Tehran North Branch, Tehran,
resistance to
sensitive to

antibiotic

Iran, in 1997. He received his PhD. in Applied Chemistry from

0.5
0

0
AgNPs

same university in 2006. Since 2006 he is chemistry faculty

Table 1. Antibacterial activity of AgNPs in dose of (5, 0.5 and 0.05 μg/mL).

member of this university and his main areas of interests are

AgNPs (μg/Ml)

Biofuels, Nano technology, Green chemistry and water

Gram positive (S.aureus)

treatment.
resistance to
sensitive to

antibiotic

Pirouz Derakhshi received M.Sc. degree in analytical chemistry

14

0
5

AgNPs

from Islamic Azad University-Tehran North Branch, Tehran, Iran,

in 1993. He received his PhD. in Applied Chemistry from same
university in 2009. Since 2009 he is chemistry faculty member
of this university and studied in Chemical Nano- Technology.
Thick nano particle

Sensitive to AgNPs
suspention

Fereshte Motiee received M.Sc. degree in analytical chemistry

resistance to

from Islamic Azad University-Tehran North Branch, Tehran,

antibiotic
30

Iran. He received his PhD. in Applied Chemistry from the

same university. He is expertise in Material Characterization,
Mechanical Properties, Thermal Analysis, Thermogravimetry,
Rubber, fillers and biofillers Polymer Chemistry, Materials
Gentamicin

Chemistry and Physical Chemistry.

antibiotic

Sensitive to

Sensitive to
antibiotic
47

27

AgNPs

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