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Open Access Full Text Article O ri g inal R esearc h

Synthesis of silver nanoparticles by using tea leaf

extract from Camellia Sinensis
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International Journal of Nanomedicine
1 August 2012
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Yuet Ying Loo 1 Abstract: The development of the biological synthesis of nanoparticles using microorganisms
Buong Woei Chieng 2 or plant extracts plays an important role in the field of nanotechnology as it is environmentally
Mitsuaki Nishibuchi 3 friendly and does not involve any harmful chemicals. In this study, the synthesis of silver
Son Radu 1 nanoparticles using the leaves extract of Chinese tea from Camellia sinensis is reported. The
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synthesized nanoparticles were characterized using UV-vis spectroscopy, X-ray diffraction

1
Centre of Excellent for Food Safety
Research, Faculty of Food Science and (XRD), transmission electron microscopy (TEM), and Fourier transform infrared (FTIR)
Technology, Universiti Putra Malaysia, spectroscopy. The XRD analysis shows that the synthesized silver nanoparticles are of face-
Selangor, Malaysia; 2Department
centered cubic structure. Well-dispersed silver nanoparticles with an approximate size of 4 nm
of Chemistry, Faculty of Science,
Universiti Putra Malaysia, Selangor, were observed in the TEM image. The application of the green synthesized nanoparticles can
Malaysia; 3Center for South East Asian be used in many fields such as cosmetics, foods, and medicine.
Studies, Kyoto University, Kyoto, Japan
Keywords: silver, nanoparticles, green synthesis, leaf extract, transmission electron microscopy,
nanotechnology

Introduction
The development of toxicity free metal nanoparticles has become a great challenge
in recent times. The primary challenge in this focal area is the maximization of the
usage of environmental friendly materials in the generation of metal nanoparticles.
Various chemical and physical methods have been employed to prepare silver
nanoparticles with different sizes and shapes, such as UV irradiation,1,2 microware
irradiation,3,4 chemical reduction,5–7 photochemical method,8,9 electron irradiation,10,11
and sonoelectrochemical method.12 However, most of the reported methods involve
more than one step, high energy requirement, low material conversions, difficulty in
purification, and hazardous chemicals. The chemical synthesis of nanoparticles may
lead to the presence of some toxic chemical species adsorbed on the surface that may
have adverse effects in its application. The synthesis of nanoparticles by green method
can potentially eliminate this problem. Therefore, there is an urgent need to develop
a green process of nanoparticle synthesis.
Green synthesis methods employing either biological microorganisms or plant
Correspondence: Son Radu extracts have emerged as a simple and alternative to chemical synthesis. Green synthesis
Centre of Excellent for Food Safety provides advancements over chemical methods as it is environment friendly, cost
Research, Faculty of Food Science and
Technology, Universiti Putra Malaysia,
effective, and easily scaled up for large scale synthesis. Generally, the green synthesis
43400 UPM Serdang, Selangor, Malaysia method involves three main steps, (1) solvent medium selection, (2) environmental
Tel +603 89468361
Fax +603 89423552
benign reducing agent selection, and (3) non-toxic substances for nanoparticles stability
Email son@putra.upm.edu.my selection.13 The synthesis of nanoparticles by using plant extracts can be advantageous

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Dovepress © 2012 Loo et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article
http://dx.doi.org/10.2147/IJN.S33344 which permits unrestricted noncommercial use, provided the original work is properly cited.
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over other biological processes because it eliminates the Erlenmeyer flask at room temperature. Any color changes of
elaborate process of maintaining cell cultures and can be the solution were observed.
suitably scaled up for large scale production under non-
aseptic environments. Characterization
Silver nanoparticles are well known as one of the most The crystallinity and phases of the Ag nanoparticles
universal antimicrobial substances in the field of biology were characterized by X-ray diffractometer (XRD-6000,
Shimadzu, Japan) with Cukα radiation (λ = 1.5412 Å) in the
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and medicine due to their strong biocidal effect against

microbial species, which has been used for centuries to range of 10°–80° with 2°/min scanning rate. The functional
prevent and treat various diseases, most notably infections.14 and composition of Ag nanoparticles were characterized by
Silver nanoparticles also reported to possess anti-fungal,15 Fourier-Transform Infrared (FTIR, Perkin Elmer, Spectrum
anti-inflammatory,16 anti-viral,17 anti-angiogenesis18 and BX) spectroscopy in the range 4000–280 cm-1. In addition,
anti-platelet activity.19 Recently, the development of silver the optical property of prepared Ag nanoparticles was
nanoparticles is expanding. They are now used as part of analyzed via UV-visible (UV-Vis, Perkin Elmer, Lambda 35)
clothing, food containers, wound dressings, ointments, and absorption double beam spectrophotometer with a deuterium
implant coatings. Some silver nanoparticles’ applications and tungsten iodine lamp in the range from 300–600 nm at
have received approval from the US Food and Drug room temperature.
Administration.20 The morphology of the prepared Ag nanoparticles was
Pu-erh teas are known as medicinal tea in China. Pu-erh observed by Transmission Electron Microscopy (TEM,
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teas are reported to have high antioxidant properties and the Hitachi, H7100). Ag nanoparticles were sonicated for 15 min
ability to decrease the cholesterol, lower blood pressure, by a sonicator (50  Hz, Soniclean). Then, the dispersed
prevent cardiovascular diseases and enhance the normal solution was dipped to a copper grid at room temperature.
flora in our gastrointestinal tract.21 In the present work, we After drying, sample was analyzed at 80 kV. The particle
have proposed a green chemical method for synthesizing Ag size distributions were determined using UTHSCSA Image
nanoparticles using pu-erh tea leaves extract from Camellia Tool Program (version 3.00; Dental Diagnostic Science,
sinensis as reducing and capping agents. To the best of UTHSCSA, San Antonio, TX).
our knowledge, this study is new and presents a simple
methodology to synthesize Ag nanoparticles efficiently at Results and discussion
room temperature. The color change was noted by visual observation in the
Erlenmeyer flask which contains AgNO3 solution with
Materials and methods pu-erh tea extract. The color of the AgNO3/tea extract
solution changed from colorless to light brown after 5 min
Materials
and eventually to dark brown (Figure 1). This color change
Pu-erh tea is traditionally made by the leaves of old wild tea
indicates the formation of Ag nanoparticles in the solution.
trees of Camellia sinensis var. assamica, which are found in
Tea extract without AgNO3 did not show any color changes.
south-western China as well as the bordering tropical regions
in Burma, Vietnam, Laos and eastern parts of India. The The formation of Ag nanoparticles was further confirmed by
leaves are artificially fermented for 6 months to a year with
microorganisms to produce pu-erh tea. Commercial pu-erh
(A) (B)
tea was purchased from local tea shop in Malaysia. Silver
nitrate (AgNO3) was purchased from Merck, Germany.

Synthesis of silver nanoparticles

The pu-erh tea leaves extract was prepared by weighing 10 g
of pu-erh tea leaves in 500 mL beaker along with 100 mL
of distilled water and maintained at 60°C for 10 min before
decanting it. The solution was filtered by 0.45 µm Milipore
membrane filter and followed by 0.2 µm Millipore membrane
filter. For synthesis of silver nanoparticles, 100  mL of Figure 1 Aqueous solution of 10−3 M AgNO3 with pu-erh tea leaves extract (A) before
AgNO3 (1 mM) was reacted with 12 mL of the tea extract in adding the tea extract and (B) after addition of tea extract at 5 minutes.

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 Dovepress Synthesis of silver nanoparticles

using UV-visible spectroscopy (UV-vis), X-ray diffraction 3

(XRD), Fourier-Transform infrared spectroscopy (FTIR) and

436 nm
transmission electron microscopy (TEM).
2

Absorbance
Figure  2  shows XRD patterns for Ag nanoparticles
synthesized by pu-erh tea leaves extract. Five main
characteristic diffraction peaks for Ag were observed at 1
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2θ = 38.4, 44.5, 64.8, 77.7 and 81.7, which correspond to

the (111), (200), (220), (311), and (222) crystallographic
planes of face-centered cubic (fcc) Ag crystals, respectively 0
300 350 400 450 500 550 600 650 700
(JCPDS 00-004-0783). No peaks from any other phase were Wavelength (nm)
observed showing that single phase Ag with cubic structure
Figure 3 UV-vis spectrum of Ag nanoparticles.
nanoparticles have been obtained directly.
In general, the width of XRD peaks is related to crystallite
size. Debye-Scherrer equation was used to determine average from the C = O stretching mode in amine I group which is
crystallite diameter from half width of the diffraction peaks: commonly found in the protein,22 indicating the presence
D = (kλ)/(β cos θ), where D is mean crystallite size of the of proteins as capping agent for silver nanoparticles which
powder, λ is the wavelength of Cukα, β is the full width at half- increases the stability of the nanoparticles synthesized.
maximum, θ is the Bragg diffraction angle and k is a constant. On the other hand, the intense and broad peak at 386 cm-1
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The (111) plane was chosen to calculate crystalline size. From corresponded to the Ag metal.
Debye-Scherrer equation, the average crystallite size of silver Transmission electron microscopy (TEM) has been
nanoparticles synthesized is found to be 3.42 nm. employed to characterize the size, shape and morphology of
Figure 3 shows the UV-vis absorption spectrum of the synthesized silver nanoparticles. The TEM image of silver
synthesized Ag nanoparticles. Silver nanoparticles have nanoparticles is shown in Figure 5A. From the image, it is
free electrons, which give surface Plasmon resonance evident that the morphology of silver nanoparticles is spherical
(SPR) absorption band, due to the combined vibration of which is in agreement with the shape of SPR band in the UV-vis
­electrons of silver nanoparticles in resonance with light wave. spectrum. Figure 5B shows the histogram of size distribution
A broad absorption peak was observed at 436 nm, which is a of silver nanoparticles. The average particles size measured
­characteristic band for the Ag. No other peak was observed from the TEM image is 4.06 nm which is in good agreement
in the spectrum which confirms that the synthesized products with the particle size calculated from XRD analysis.
are Ag only. The environmentally benign silver nanoparticles
FTIR measurement was carried out to identify the possible synthesis process have potential applications in verious
biomolecules responsible for capping and reducing agent for fields. Silver nanoparticles can be applied in coating for
the Ag nanoparticles synthesized by tea leaf extract. Three solar energy, bio-labelling, food packaging, antimicrobial
obvious infrared bands are observed at 3271 cm-1, 1637 cm-1 agent and drug delivery.23,24 Several studies on the usage of
and 386 cm-1 (Figure 4). The intense broad band at 3271 cm-1 metal nanoparticles in the water filter have been carried out
is due to N−H and O−H stretching mode in the linkage of due to its antibacterial and as pesticide removal properties.25
the proteins. The medium intense band at 1637 cm-1 arises Besides that, silver nanoparticles play an important role in
(111)

Transmittance (%)
Intensity (au)

1637
(200)

(220)

(311)

3271
(222)

386

20 30 40 50 60 70 80 90 4000 3000 2000 1000 0

2θ (°) Wavenumber (cm−1)

Figure 2 XRD patterns of Ag nanoparticles. Figure 4 FTIR spectrum of Ag nanoparticles.

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A B 18
Mean = 4.06 nm
16

14

12

Frequency
10

8
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0
0 1 2 3 4 5 6 7 8 9

50 nm
Particles size (nm)

Figure 5 (A) TEM image and (B) particles size distribution of Ag nanoparticles synthesized by pu-erh tea leaves extract.

the medical area. Silver nanoparticles act as biomarker in 3. Yin H, Yamamoto T, Wada Y, Yanagida S. Large-scale and size-con-
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