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GREEN SYNTHESIS OF SILVER NANOPARTICLES FROM AQUEOUS STEM

EXTRACT OF CERIOPS TAGAL

Article  in  International Research Journal of Pharmacy · January 2018

DOI: 10.7897/2230-8407.0812258

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INTERNATIONAL RESEARCH JOURNAL OF PHARMACY

www.irjponline.com
ISSN 2230 – 8407

Research Article
GREEN SYNTHESIS OF SILVER NANOPARTICLES FROM AQUEOUS STEM EXTRACT OF
CERIOPS TAGAL
Larkins Ramteke *, B.L. Jadhav, Poonam Gawali
Department of Life Sciences, University of Mumbai, Kalina Campus, Santacruz- E, Mumbai, India
*Corresponding Author Email: [email protected]

Article Received on: 13/12/17 Approved for publication: 22/12/17

DOI: 10.7897/2230-8407.0812258

ABSTRACT

Biosynthesis of reliable and eco-friendly nanoparticles is an important branch of green nanotechnology. Ceriops tagal, a mangrove plant has various
medicinal properties. Synthesis of C.tagal mediated nanoparticles may further enhance its medicinal potency. The aqueous stem extract of C.tagal
was used as a bio-reducing agent for synthesis of silver nanoparticles Reduction of metal ions was confirmed by Ultraviolet-visible absorption
spectroscopy, transmission electron microscopy, energy dispersive spectroscopy, and x-ray diffraction. Before and after bio-reduction changes in
functional groups of plant extract was recorded by FTIR spectra. The kinetics of particle formation was dependent on time, temperature and metal salt
concentration. UV-Visible spectroscopy demonstrated peak at 455 nm. TEM and XRD results revealed shapes, size and structure of synthesized
nanoparticles. These results have indicated the formation of silver nanoparticles.

Keywords: Mangroves, Ceriops tagal, aqueous extract, silver nanoparticles.

INTRODUCTION the major species is C.tagal. Among the different mangrove

plants, the chemical constituents and bioactivities of Ceriops
Nanotechnology has wide applications in almost every field tagal have been studied extensively. Dolabrane diterpenes from
with the most exploitable properties being mechanical, thermal, C. tagal has exhibited significant anti-tumor effect15. Bark of C.
chemical, electrical, optical, magnetic, specific surface area etc 1. tagal is a powerful astringent and is used for treating
It is playing an important role in global manufacturing along hemorrhage in defecation and malignant ulcers whereas leaves
with rapid commercialization. Beside medical and are used to heal paludism and malaria; ethanolic extract of stem
pharmaceutical applications, noble metals nanoparticles are and twigs has strong feeding deterrent activity against T.
widely applied in shampoos, soaps, detergent, shoes, cosmetic castaneum adults16. Few of the compounds isolated from
products, and toothpaste2. Silver has been used as a healing and ethanolic embryo extract were effective to inhibit proliferation
antibacterial agent since ages3. Nano silver has found and growth of H-7402 and Hela cells17. It has been reported to
application in diverse areas like catalysis, textile engineering, contain condensed and hydrolysable tannins, aliphatic
water treatment, therapeutics etc furthermore it is used as a carboxylic acids, indole alkaloids, polyphenols, proteins,
strong antimicrobial, antifungal and anti-inflammatory agent4. It tannins, fatty acids, hydrocarbons, inorganic salts, inositols,
has been used in the formulation of dental resin composites5,6,7 steroids, carotenoids, chlorophyll a, b, a+b, etc18. Thus it is a
and ion exchange fibers8 and in coatings of medical rich source of bio-reductant and stabilizers. There are reports of
devices9,10,11. Nanocrystalline silver dressing is effective for monometallic nanoparticles synthesis from mangrove species-
treating wounds especially burns and chronic wounds which Rhizophora mucronata19 Avicennia marina20 Sonneratia
slowly releases small and highly reactive silver particles12. apetala21 etc. Here in, we report for the first time synthesis of
Biological effectiveness of nanoparticles is directly proportional silver nanoparticles employing aqueous stem extract of C. tagal
to its surface area. Nanoparticles exhibit new and improved (CTSE) by reduction of aqueous Ag+ ions. We also investigated
properties based on their size and shape. Chemical synthesis of effects of time course, temperature and metal salts on the rate of
nanoparticles possesses the potential hazards like synthesis.
carcinogenicity, genotoxicity, cytotoxicity, and general
toxicity13. Therefore, there is a current drive to produce ‘green’ MATERIALS AND METHODS
nanoparticles from plants because of its eco friendliness, energy Plant collection and extract preparation
efficiency and convenience. Mangroves are salt tolerant plant
species surviving under stressful conditions such as high tides C.tagal stem was collected from the Gorai creek, Mumbai,
and salinity, extreme temperatures, anaerobic & unstable India. It was chopped, dried at 40º C and pulverized. Ultra pure
substrates thus forming unique environments and floral-faunal water produced by MilliQ system was used throughout the
assemblages. India has a total mangrove cover of only 4,628 experiment. Aqueous stem extract was prepared by soaking 5 g
km2, 3% of the global mangrove area and 8% of Asia’s of stem powder in 100ml MilliQ water, for 5 min and then the
mangroves14. Around 20 out of the 35 species of true mangroves mixture was boiled at 100º C for 5 min. The freshly prepared
found in India have been identified along the Maharashtra coast extract was obtained by filtering it through Whatman filter paper
and 15 species of these are found in Mumbai; in which one of No.1 and used for further study.

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Synthesis of silver nanoparticles concentration resulted in precipitation. Maximum bio-reduction

of Ag+ to Ag0 was achieved at RT.
Synthesis of monometallic nanoparticles was initiated by
adding 5ml CTSE in 95 ml aqueous AgNO3 solution. The TEM and energy dispersive spectroscopy measurements
reaction was carried out at static condition. Reduction of Ag+
was monitored as a function of time by measuring UV-Vis TEM micrograph confirmed the synthesis of monometallic
spectra using UV-1650CP Schimadzu spectrophotometer nanoparticles. For elemental analysis of nanoparticles, a
operated at 1nm resolution. Effect of temperature on rate of signature spectrum corresponding to metal atom in the
synthesis was studied by carrying out the reactions at Room nanoparticles was obtained by EDS (Figure 1b). Weak signals
temperature (RT), 40ºC, 50ºC and 60ºC in water bath. from oxygen, chloride, carbon etc were also seen which may
Concentration of metal salt varied from 1mM – 5mM for have originated from the bio-molecules bound to the surface of
silver nitrate. the nanoparticles, while copper peaks may have origin from the
copper grid. TEM diffraction pattern of nanoparticles is shown
TEM and energy dispersive spectroscopy measurements in Figure 1c. All monometallic nanoparticles synthesized were
almost spherical with a small percentage being ellipsoidal
Size and surface morphology of bio reduced nanoparticles was (Table 2). From the few particle images we found that the size
determined by transmission electron microscope (TEM, Tecnai (diameter) varied from 5nm to 22nm for silver nanoparticles
12 Cryo, FEI, Eindhoven, The Netherlands) in the Department synthesized employing AgNO3 (Figure 1d). It was clear from
of Physics, Savitribai Phule Pune University, Pune as follows; the TEM measurements that the biosynthesized nanoparticles
Nanoparticle solution was drop coated on copper TEM grids, were poly-dispersed. TEM micrograph also indicates that the
after which film was allowed to stand for 2 min and excess nanoparticles are capped by a thin layer which is supposed to be
solution was blotted. The grid was dried properly prior to the organic material from CTSE.
measurement. An energy dispersive spectrum was recorded with
the same instrument at the energy range 0-20 keV. XRD

X-ray diffraction measurements The crystalline nature of nanoparticles was confirmed by X-ray
diffraction analysis. The crystallite size of the nanoparticles was
After complete reduction of metal ions by extracts, the solutions estimated from the Debye–Scherrer formula22.
were centrifuged at 10,000 rpm for 15 min at RT. The pellet
obtained was re-dispersed and centrifuged with MilliQ water. d = 0.9λ/β cos θ
This process was repeated three times to get rid of any free
entities. Phase formation of nanoparticles was studied by Where 0.9 is the shape factor, λ is x-ray wavelength, 1.54 Å, β
preparing thin film of thoroughly dried nanoparticles on glass full width at half the maximum intensity in radians, and θ is the
slides. Diffraction data was recorded on Schimadzu XRD 7000 Bragg angle.
diffractometer with Cu Kα radiation (1.54 Å) source operating at
40 kV voltage and a 30 mA current at Department of Chemistry, Strong Bragg reflections were obtained corresponding to (111),
University of Mumbai, Mumbai. (200) and (220) planes for silver nanoparticles, which are the
principal diffracting planes of face centered cubic symmetry
Fourier Transform Infrared (FTIR) Spectroscopy (Table 2). High degree of crystallinity of the nanoparticles was
reflected by the intensity of peaks. The crystallite size of silver
FTIR spectrum of extracts before and after reduction of metal nanoparticles synthesized using AgNO3 (Figure 1 e) was ∼6 nm.
ions was obtained using FTIR (Perkin Elmer) Frontier
spectrophotometer at Department of Chemistry, University of FTIR
Mumbai, Mumbai. 5% plant extract and supernatant of bio-
reduced samples was subjected to IR source 400 cm−1 - 4000 FT-IR spectroscopic studies were carried out to identify the
cm−1. functional group involved in capping and efficient stabilization
of the metal nanoparticles. Representative absorption peaks of
RESULTS the nanoparticles obtained in the present study is presented in
Visual observations and UV-Visible spectroscopy Table 4. The plant extract shows intense broad stretching at
3550-3200 cm-1 which arises due to the free O-H groups
Bio-reduction of salt to respective metal ions in presence of present in alcohols and phenols 22, weak stretch at 2260 – 2100
extract was monitored as a function of time using UV-Visible cm-1 is due to C≡C from alkynes while the IR peak at 1690-1630
spectroscopy. The surface plasmon resonance (SPR) peak of cm-1 could be assigned to characteristic asymmetrical stretch of
absorption spectra for CTSE was found at 455nm for carboxylate group23. The shifting of peaks occurred after
nanoparticles synthesized using AgNO 3 (Figure 1a) salt solution synthesis of silver nanoparticles (Figure 1f). Based on the band
(Indian patent application 1320/MUM/2013 A). The well- shift occurring at hydroxyl and carbonyl groups it can be
known intense brown colour was observed after silver concluded that both hydroxyl and carbonyl groups of C.tagal are
nanoparticles formation. Complete reduction of Ag+ metal ions involved in the synthesis of nanoparticles. As stated earlier,
was done in 120 min, while the colour change was observed CTSE is mainly composed of terpenoids, flavonoids, alkaloids
after 15 min. The pH of CTSE was 7. Optimization of different and ployphenols which may play an important role in reduction,
concentration of metal salts against kinetics of reaction was stabilization and assembly of synthesized nanoparticles.
observed (Table 1). The rate of synthesis was highest at 2mM
for silver nitrate solution. The lower salt concentration showed
comparatively low rate of synthesis whereas higher salt

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Figure 1a: UV visible spectra recorded as function of reaction time of Figure 1 b: Representative spot EDS profile confirming the presence
2mM AgNO3 solution with CTSE at RT. of silver nanoparticles biosynthesized by challenging CTSE with
AgNO3 salt.

Figure 1 c: TEM Diffraction pattern of silver nanoparticles. Figure 1.d: TEM micrograph of silver nanoparticles biosynthesized
by challenging CTSE with AgNO3 salt

Figure 1e: Representative XRD profile of silver nanoparticles. Figure 1.f: FTIR absorption spectra of CTSE before bioreduction
(a), after complete bioreduction of AgNO 3 salt (b)

Table 1: Optimization of nanoparticle synthesis

Sr. No Salts in mM conc. Synthesis time in min. Temp. Colour λ max

ºC
1. 2mM AgNO3 15-30 RT Intense brown 455 nm
(38º C)

Table 2: Size, shape and yield of NPs synthesized from C.tagal stem

Sr. No. NPs Size in nm Shape Yield mg/mL

1. Silver NPs from silver nitrate 5-22 Predominately Spherical, few ellipsoidal 620

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Table 3: XRD details of silver nanoparticles

Sr.No. Nanoparticles Crystal 2Ɵ of the hkl Crystallite size

synthesized from structure intense peak
salt (deg)
1. AgNO3 FCC a. 27.756 (111) ∼6 nm
b. 32.174 (200)
c. 46.300 (220)

Table 4: FTIR details of CTSE and bio-reduced AgNo3

Peaks and functional groups Free O-H groups from alcohols and phenols 3550 C≡C from alkynes Asymmetrical stretch of carboxylate
-3200 cm-1 2260 – group 1690- 1630 cm-1
2100 cm-1
C.tagal stem extract 3306.66 2119.25 1634.46
Bio-reduced AgNo3 3271.40 2103.33 1634.52

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