Green Synthesis of Silver Nanoparticles Using Cressa Cretica Leaf

Transcription

International Journal of Advanced Chemical Science and Applications (IJACSA)
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Green Synthesis of Silver Nanoparticles Using Cressa Cretica Leaf
Extract and its Antibacterial Efficacy
1
S. Balasubramanian, 2U. Jeyapaul, 3A.John Bosco, 4S.Mary Jelastin Kala
1
Department of Chemistry, St.xaviers College, Palayamkottai, Tirunelveli-627002, India
Department of chemistry, St.xaviers College , Palayamkottai , Tirunelveli-627002 , India
3
Department of chemistry ,SRM University, Faculty of Engineering and Technology, Kattankulathur, Kancheepuram603 203, India
Email : 1bensosurya@yahoo.com
2,4.
[Received:26th March 2015;
Accepted:7th April 2015]
Revised: 6th April 2015;
Abstract : Nanomaterials are widely used in a variety of
fields. Nanomaterials are commonly synthesized using
physical, chemical and biological methods. The biological
green synthesis methods are rapid, cost effective and ecofriendly . The present investigation deals with biosynthesis
of silver nanoparticles from silver nitrate using cressa
cretica leaf extract. The biosynthesized silver nanoparticles
were characterized by UV-Visible spectrophotometer, XRay Diffraction, FTIR spectra, EDX and Scanning
Electron Microscopy. The antibacterial activity of
biosynthesized silver nanoparticles was investigated by
diffusion disk method. It is suggested that the biologically
synthesized silver nanoparticles are more effective against
various disease causing pathogens.
Key words: silver nanoparticles , cressa cretica , uv-visble
spectra, XRD , antibacterial activity
I. INTRODUCTION
Plants have flavonoids, alkaloids and polyphenolic
compounds which may reduce the silver ions to silver
nanoparticles and acts as capping and stabilizing agent
[9].
The silver nanoparticles are highly toxic to several
pathogenic organisms and hence play a vital role in
treatment of many diseases [10,11].Silver has long been
recognized as having an inhibitory effect towards many
bacterial strains and microorganisms [12]. Antibacterial
activity of the silver containing materials is used in
medicine to reduce infections in burn treatment [13] and
arthroplasty [14], as well as to prevent bacteria
colonization on prostheses [15], catheters [16],vascular
grafts, dental materials [17], stainless steel materials
[18], and human skin [19]. Silver nanoparticles also
exhibit a potent cytoprotective activity towards HIVinfected cells[20].
Cressa
cretica
(Linn)
belonging
to
family
Convolvulaceae, commonly known as Rudravanti is a
erect, small, dwarf shrub, [21] usually grows in sandy or
muddy saline habitats [22]. Cressa cretica is a
remarkable salt tolerant plant, common in coastal areas
[23] usually occurring in mono specific stands along the
landward edge of marshes [24] .This plant is distributed
throughout India, Timor, and Australia (Western
Australia, Northern Territory, Southern Australia,
Queensland, New South Wales, Victoria). The plant is
used as antibilious, antitubercular ,expectorant [25]-[26],
antihelmintic, stomachic, tonic and aphrodisiac
purposes,and enriches the blood and is useful in treating
constipation, leprosy, asthma, urinary discharges, and as
an appetizer[27]. Dry leaves of Cressa cretica crushed
with sugar are used as emetic in Sudan[28]
Nanotechnologies are among the fastest growing areas
of scientific research and have important applications in
a wide variety of fields. Nanomaterials have different
physical, chemical and biological properties than their
macro scaled counterparts [1]-[2]. Nanoparticles are
commonly synthesized using two steps: top-down and
bottom-up [3]. The bulk materials are gradually broken
down to nanosized materials in top-down approach.
Atoms or molecules are assembled to molecular
structures in nanometer range in bottom-up approach
[4]. In bottom-up approach, silver nanoparticles are
synthesized by various chemical, physical and biological
methods. Biological synthesis of nanoparticles is gaining
importance because it is safe, cost-effective,
biocompatible, non-toxic and eco-friendly whereas
physical and chemical processes are expensive, toxic
and hazardous to environment [5]. The natural sources
like plants and microorganisms are used for synthesizing
silver nanoparticles by biological approach [6].
Synthesis of nanoparticles using microorganisms
consume more processing time, whereas using the plant
extract mediated methods require less processing time
[7] and also suitable for large scale production [8].
Fig:1 Cressa cretica
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ISSN (Print):2347-7601, ISSN (Online): 2347-761X, Volume -3, Issue -1, 2015
65
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Vernacular names[29]-[32]
Hindi – Rudravanti
Tamil – Uppu Marikkozhundu
Telugu – Uppusanaga
Sanskrit – Rudanti
Bengali – Rudravanti
Cressa cretica leaves contain bioactive constituents such
as carbohydrates, flavonoids, phytosterols, terpenes,
tannins, glycosides, fixed oil and sugars[33,34]
Based on the importance of green synthesis of silver
nanoparticles from leaf extract, the present study was
carried out to synthesize and characterize the silver
nanoparticles using Cressa cretica (Linn) leaf extract.
II. MATERIALS AND METHODS :
2.1.Collection of plant samples:
Cressa cretica plants were collected from Tuticorin
District, Tamilnadu, India. The plant leaves were
thoroughly washed thrice with tap water and then with
double distilled water to remove dust particles, air-dried
for a week under shade at room temperature, finely cut,
milled into a fine powder and was stored in airtight
containers for later analysis.
2.2.Preparation of aqueous plant leaf extracts:
15 gm of the leaf powder was mixed well with 100 ml of
double-distilled water and boiled at 60 0 C for 30 min.
After boiling, the extract was filtered through Whatman
No.1 filter paper. The supernatant was collected and
stored at 4 ºC for further nanoparticles process[35].
2.3.Synthesis of silver nanoparticles
1mM aqueous solution of Silver nitrate (AgNO3) was
prepared and used for the synthesis of silver
nanoparticles. 1 ml of Cressa cretica leaf extract was
added into 100 ml of aqueous solution of 1 mM silver
nitrate for reduction of Ag+ ions and kept at room
temperature for 24 hours[36]. Formation of reddish
brown colour confirmed the silver nanoparticles.
III. CHARACTERIZATION STUDIES:
The green synthesized silver nanoparticles were
characterized by the following methods:
3.1.Visual Observation:
A change of colour from pale yellow to reddish brown
was observed in the solution after visible irradiation.
3.2.UV Spectrophotometric analysis:
3.3. Fourier Transform Infra Red Spectroscopy
Analysis:
The functional groups in the biosynthesized Ag NPS
solution were analyzed by FTIR spectroscopy. These
measurements were carried using a perkin Elmer
spectrum RX I FTIR instrument with a wavelength
range of 4000 to 400 nm .The results were compared for
shift in functional peaks.
3.4.Field Emission Scanning Electron Microscopy :
FESEM was used to characterize the mean particle size,
morphology of the AgNPs. A small drop of
biosynthesized Ag NPS solution was placed on glass
slide and allowed to dry. The samples were analyzed by
using FEI Quanta 200 FEG machine at a low vacuum
in the range 10-20 Kv
3.5. Energy Dispersive X-ray Analysis:
The elemental composition of the synthesized
nanoparticles was analyzed with energy dispersive
spectroscopy coupled to scanning electron microscope.
3.6.XRD analysis :
The silver nanoparticles solution thus obtained was
purified by repeated centrifugation at 10,000 rpm for 20
min. The purified Ag nanoparticles are dried. The
structure and composition of Ag nanoparticles were
studied by XRD (XPERT-PRO Machine). The data was
collected in the 2Ө range. The crystalline domain size
was calculated from the width of XRD peaks using
Scherrer’s equation.
Dabye- Scherrer’s equation, D = K λ/ β Cos Ө ;
Where, D = average crystalline domain size; β is the
Full Width at Half Maximum (FWHM), K= 0.94, λ
= wave length of X ray, and Ө is the diffraction angle.
3.7. Antibacterial activity :
The antimicrobial activity of AgNPs was estimated
against pathogenic bacteria such as Enterococcus
faecalis, Staphylococcus aureus (gram- positive
bacterias), Pseudomonas aeruginosa, E. coli (gramnegative bacterias) by disc diffusion method[37].The
bacterial cultures were grown in Brain Heart Infusion
liquid medium at 37 °C. After 12 hrs of growth, each
microorganism, at a concentration of 1 x 106 cells/mL
equivalent to 0.5 Mc Farland Standard was spread on the
surface of Mueller-Hinton agar plates. The dilutions
were made in sterile low glucose Nutrient broth. Test
pathogens were spread on the test plates- Muller Hinton
agar for bacteria. The synthesized Ag nanoparticles and
positive control were loaded onto 6 mm diameter sterile
discs and placed in the plates respectively. After 24hrs
of incubation, the zone of inhibition (mm in diameter)
was measured and taken as the activity against the test
pathogen.[38]
The formations of leaf extract mediated silver
nanoparticles were confirmed by the spectral analysis.
The UV spectra of the biosynthesized silver
nanoparticles were recorded using shimadzu UV-1800
Spectrophotometer by continuous scanning from 200nm
to 900nm and distilled water was used as the reference
for the baseline correction.
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66
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Fig -3: UV Spectral analysis of silver nanoparticles
synthesized from cressa cretica leaf extract
IV. RESULTS AND DISCUSSION :
4.1. UV- Visible Spectroscopy analysis:
The absorption spectrum of Ag NPs from cressa cretica
leaf extract is shown in fig.3. A well defined peak at
444.5nm exhibited by the nano metallic Ag particles and
broadening of peak indicated that the particles are
polydispersed[39]-[40]. The colour change from the
colourless to reddish brown due to the excitation of free
electrons in the nanoparticles[41].It confirmed the
successful synthesis of Ag NPs .
The nanoparticles were primarily characterized by UVVis spectroscopy for the analysis of nano particles.
4.2. FT-IR analysis:
Fig.4. shows the FT-IR spectrum of synthesized Ag NPS
from leaf extract of cressa cretica. FT-IR measurement
was carried out to identify the possible bio molecules
responsible for capping and reducing agent for the
AgNPs synthesized by cressa cretica leaf extract. The
peak at 3437.43 cm-1 corresponds to O-H stretching of
phenols and alcohols and N-H stretching of amines. The
peak at 2360.37 cm-1 corresponds to C≡N stretching of
nitriles and C≡C stretching of alkynes , the peak at
2068.86 cm-1 corresponds to CO2 molecule ,the peak at
1643.80 cm -1 corresponds to C═O stretching of
aromatic ester and enolic ketones , the peak at 1312.60
cm-1 corresponds to C-O stretching of ester and phenols
and C-N stretching of amines, peaks at 671.40 cm-1
corresponds to C-Cl stretching of aryl halides. These
bio molecules reduced Ag + to Ag as well as stabilizing
Ag NPs.
Fig:2 Colour change indicating formation of Ag NPS
Fig-2: solution of (A) Cressa cretica leaf extract (B)1m
M AgNO3 Solution without leaf extract (c) 1m M
AgNO3 Solution with leaf extract after 5 mins (D) 1m
M AgNO3 Solution with leaf extract after 24hrs
ACIC
St.Joseph's College ( Autonomous)
Trichy-2
Spectrum Name: Bot-Liq-1.s p
100.0
95
90
2360.37
1312.60
85
2068.86
80
75
70
671.40
65
60
55
50
%T
45
1643.80
40
35
30
25
20
15
3437.43
10
5
0.0
4000.0
3600
3200
2800
2400
2000
1800
1600
1400
1200
1000
800
600
400.0
cm-1
Fig - 4 : FT-IR Spectra of silver nanoparticles synthesized from cressa cretica leaf extract
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67
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4.3. XRD Analysis:
Fig-5 : XRD patterns of silver nanoparticles using cressa cretica leaf extract
The green synthesized silver nanoparticles are highly crystalline with diffraction peaks could be obviously assigned to
the face-centered cubic phase of metallic silver. Fig. 5 shows main characteristic diffraction peaks for Ag observed at
2θ values of 38.03°, 44.46°, 64.65°and 77.32° are indexed to the (100), (200), (220)and (311) reflections of the fcc
structure of metallic silver respectively. The average grain size of the silver nanoparticles formed in the bio reduction
process was determined using Scherer’s formula.
4.4. SEM and EDX analysis:
Fig-6 : SEM image of Ag nanoparticles using cressa cretica leaf extract
Fig. 6 shows the morphology investigated using
scanning electron microscopy (SEM). This picture
reveals that the green synthesized silver NPs are
approximately rod like structure and the particles are
slightly agglomerated and the dimensions of nanorods
are between 50–200 nm in width and few microns in
length. The results of energy-dispersive spectroscopy
(EDX) analysis are shown in Fig. 7. It is confirmed that
the significant presence of elemental silver, which
indicates bio reduction of silver ion to elemental silver.
The strong signal in the silver region was observed at 3
keV for silver nanoparticles due to the Surface Plasmon
Resonance [42]-[43].
Fig-7 : EDX image of Ag nanoparticles using cressa cretica leaf extract
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68
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4.5.Anti bacterial Activity:
Silver is well known as one of the most universal
antibacterial substances. Antibacterial activity is
investigated
against
Enterococcus
faecalis,
Staphylococcus aureus (gram- positive ), Pseudomonas
aeruginosa, E. coli (gram-negative) by disc diffusion
method are shown in Fig. 8.[44]. The diameter of
inhibition zones (mm) around each well with silver
nanoparticles solution are represented in Table 1. The
table.1. shows that the zone of inhibition (ZOI) was
increased when increasing the concentration of Silver
nanoparticles[45]. The maximum antibacterial activity
in 500µg/ml concentration of the synthesized Ag
Table : 1 Zone inhibition of Silver nanoparticles
Serial.
No
1
2
3
nanoparticles was 14 mm for pseudomonas aeruginosa ,
11mm for E. coli, 10 mm for staphylococcus aureus and
14 mm for Enterococcus faecalis receptively. The silver
nanoparticles synthesized by cressa cretica leaf extract
are found to have highest antibacterial activity against
pseudomonas aeruginosa (14 mm) and Enterococcus
faecalis (14 mm) respectively and the lesser
antimicrobial activity of silver nanoparticles is found
against staphylococcus aureus(10 mm).The antibacterial
activity of Ag NPS studies confirmed that the Ag NPs
synthesized by leaf extract of cressa cretica exhibited
good antibacterial potential against gram-positive and
gram-negative bacterial strains.
Zone of inhibition of Ag NPS ( mm)
Strains Pseudomonas Staphylococcus
aeruginosa
aureus
µg/ml
500
14
10
250
10
125
10
-
E.coli
Enterococcus
faecalis
11
-
14
-
Fig. 8 : Antibacterial activity of siver nanoparticles synthesized by cretica leaf extract against human pathogens
V.CONCLUSION :
The present study concluded that the leaf cressa cretica
can be used as an excellent source for synthesizing the
silver nanoparticles. The primary confirmatory for the
silver nanoparticles was colour changes and Uv-Vis
absorption spectra of silver nanoparticles forming peak
at 444.5 nm. The SEM image confirmed that the green
synthesized silver nanoparticles are rod in shape. The
green synthesized nanoparticles have more effective
antibacterial activity to the pathogens. The present study
emphasizes the use of medicinal plants for synthesis of
silver nanoparticles with potent antibacterial effect. The
Applications of such eco-friendly nanoparticles in
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69
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bactericidal, wound healing and other medical and
electronic applications, makes this method potentially
exciting for the large-scale synthesis of other inorganic
materials (nanomaterials).
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[12]
The authors thank the
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Green Synthesis of Silver Nanoparticles Using Cressa Cretica Leaf

Transcription

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