Academic Sciences Asian Journal of Pharmaceutical and Clinical Research

Vol 5, Suppl 1, 2012 ISSN - 0974-2441

Vol. 4, Issue 3, 2011 Research Article

ISSN - 0974-2441
GREEN SYNTHESIS AND CHARACTERIZATION OF NANO SILVER USING LEAF EXTRACT OF
MORINDA PUBESCENS
JANCY MARY E AND INBATHAMIZH L*
Department of Biochemistry, Meenakshi College for Women, Chennai, Tamilnadu, India, E mail: [email protected]
Received: 08 November 2011, Revised and Accepted: 11 January 2012
ABSTRACT
Wide application of nanoparticles stimulates the need for synthesizing them. But, the conventional methods are usually hazardous and energy
consuming. This leads to focus on “Green synthesis” of nanoparticles which seems to be an easy, efficient and eco-friendly approach. In this study,
the green synthesis of silver nanoparticles was carried out using leaf extract of Morinda pubescens as reducing agent. The nano synthesis was
monitored under different range of temperatures, radiations, time periods and concentrations of the leaf extract. The optimized nano silver thus
obtained was quantified and characterized using UV-Visible spectroscopy, Scanning Electron Microscopy, Transmission Electron Microscopy, X-ray
Diffraction and Fourier Transform Infrared spectroscopy. The results emphasized the potent application of Morinda pubescens leaves in the
synthesis of nano silver with economic viability and ease in scaling up for mass production.
Key words: Nanoparticles, Green synthesis, Morinda pubescens, UV-Visible spectroscopy, Scanning Electron Microscopy, Transmission Electron
Microscopy, X-ray Diffraction, Fourier Transform Infrared spectroscopy.
INTRODUCTION
Synthesis of nanoparticles to have a better control over particle size, The present study deals with the successful synthesis of silver
distribution, morphology, purity, quantity and quality, by employing nanoparticles from silver nitrate through a simple green route using
environment friendly economical processes has always been a the leaf extract of Morinda pubescens as the reducing agent.
challenge for the researchers1. Chemical reduction is the most
frequently applied method for the preparation of silver MATERIALS
nanoparticles as stable, colloidal dispersions in water or organic Silver nitrate solution was purchased from HiMedia Laboratories
solvents. Commonly used reductants are borohydride, citrate, Pvt. Limited, Mumbai, India. The Leaves of Morinda pubescens Linn.
ascorbate, and elemental hydrogen2. The synthesis of nanoparticles were collected from the forest region of IIT Campus, Chennai and
by chemical reduction methods are often performed in the presence identified by the Department of Plant Biology and Plant
of stabilizers in order to prevent unwanted agglomeration of the Biotechnology, Meenakshi College, Chennai.
colloidal nanoparticle solution. Also, some chemical methods cannot
avoid the use of toxic chemicals in the synthesis protocol. Since METHODS
noble metal nanoparticles such as silver nanoparticles are widely
Preparation of aqueous silver nitrate
applied to human contacting areas, there is a growing need to
develop environmentally friendly processes for their synthesis that 1 mM Silver nitrate solution was prepared and stored in amber
do not use toxic chemicals. Biological methods of nanoparticle coloured bottle.
synthesis using microorganism3, enzyme4 and plant or plant extract5
have been suggested as possible ecofriendly alternatives to chemical Preparation of extract by conventional method
and physical methods. Using plant for nanoparticle synthesis can be
The Morinda pubescens leaves were washed several times with
advantageous over other biological processes as it eliminates the
deionised water. 100gm of finely cut Morinda pubescens leaves was
elaborate process of maintaining cell cultures. It can also be suitably
taken and boiled in 300ml of double distilled water for 3mins and
scaled up for large-scale synthesis of nanoparticles.
filtered. After centrifugation at 10,000rpm for 15mins, the
Silver nanoparticles have more applications in many areas, including supernatant was collected and stored at 4°C.
biomedical, materials science, and catalysis. A single silver
Preparation of extract by homogenization method
nanoparticle interacts with light more efficiently than a particle of
the same dimension composed of any known organic or inorganic The Morinda pubescens leaves were washed several times with
chromophore6. Silver is also the only material whose plasmon deionised water. 100g of the leaves was homogenized in 300ml
resonance can be tuned to any wavelength in the visible spectrum. water with the help of mortar and pestle and filtered. Then the
Fundamental studies carried out in the last three decades show that filtrate was centrifuged for 15min at 10,000rpm, supernatant was
silver nanoparticles exhibit a rare combination of valuable collected and stored at 4°C.
properties, namely, unique optical properties associated with the
surface Plasmon resonance (SPR), well-developed surfaces, catalytic Optimization and synthesis of silver nanoparticles
activity, high electrical double layer capacitance, etc 7. Nanosilver Different concentrations of leaf extracts (1ml, 3ml and 5ml) were
has been used extensively as anti-bacterial agent in the health taken separately and to this 10ml of 1mM Siver nitrate solution was
industry, food storage, textile coatings and a number of added with constant stirring and exposed to different conditions like
environmental applications. It is important to note that despite of sunlight radiation,UV radiation, several short burst of microwave
decades of use, the evidence of toxicity of silver is still not clear. irradiation at a frequency of 2.45GHz in a domestic microwave oven
Products made with nanosilver have also been approved by a range (National Model N N-GD 576M) , at a power output of about 100W in
of accredited bodies8. a cyclic mode (on 15s, off 15s) to prevent overheating as well as
Morinda pubescens commonly known as Indian mulberry or Noni or aggregation of metals, Room temperature and Direct boiling. The
Bartondi belongs to family Rubiaceae. The plants are found as weeds colour change of the solution was checked periodically. The colour
in the dried region of Maharashtra9. The ethnobotanical uses of M. change of the leaf extract from yellow to dark brown indicated the
pubescens have been reported10. The bark of M. pubescens is useful in silver nanoparticles were synthesized from the leaves. Bioreduction
eczema, fever due to primary complex, ulcer and glandular swellings of silver ions in the solution was monitored using Genesys 10 UV-VIS
while leaves are useful in digestive disorders especially in children spectrophotometer. Then the conical flasks were incubated at room
and in venereal diseases. temperature for 48 hours. The contents were centrifuged at
10,000rpm for 15 minutes. The supernatant was used for the
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Asian J Pharm Clin Res, Vol 5, Suppl 1, 2012, 159-162

characterisation of the silver nanoparticles. The same was electrons that are transmitted through the sample.
performed for both conventional and homogenization method of
extracts. Using Philips CM12 Transmission Electron Microscope, electrons in
the electron gun were accelerated across a potential difference of the
Production and Recovery of silver nanoparticles by order of 100,000 volts between the cathode and anode. The
centrifugation condenser lens was used to focus the electron beam emerging from
the electron gun onto the specimen to permit optimal illuminating
Among various concentrations and methods used, sunlight conditions for visualizing and recording the image. The specimen
irradiation method was very effective and 1ml of homogenized leaf image generated by the objective lens was subsequently magnified
extract had shown more synthesis of nanoparticles. Further it was in one or two more magnification stages by the intermediate and
chosen for bulk production as 10ml leaf extract in 100ml of 1mM projector lens and projected onto a photographic plate.
Siver nitrate. After bioreduction, the solution consisting of hydrosols
of silver nanoparticles was subjected to centrifugation at 10,000rpm Characterization of silver nanoparticles by X-Ray Diffraction
for 15 minutes. The pellet formed was dissolved in 0.1 ml of toluene (XRD)
water and air dried.
The air dried nanoparticles were coated onto XRD grid and analyzed
Characterization of silver nanoparticles by UV-Visible for the formation of silver nanoparticle by Philips X-Ray
spectroscopy Diffractometer with Philips PW 1830 X-Ray Generator operated at a
voltage of 40kV and a current of 30mA with Copper Potassium alpha
UV-Visible spectroscopy is used for monitoring the signature of radiation. The diffracted intensities were recorded from 10' to 80' of
silver nanoparticles. UV-Visible spectroscopy is a powerful tool for 20 angles.
the characterization of colloidal particles. Noble metal particles are
ideal candidates for study with UV- Vis spectroscopy, since they Characterization of silver nanoparticles by Fourier Transform
exhibit strong surface plasmon resonance absorption in the visible Infrared spectroscopy (FTIR)
region and are highly sensitive to the surface modification.
The dried silver nanoparticles were subjected to FTIR analysis by
Characterization of silver nanoparticles by Scanning Electron Potassium Bromide pellet (FTIR grade) method in 1: 100 ratios and
Microscopy (SEM) spectrum was recorded in Nicolet Impact 400 FT-IR
Spectrophotometer using diffuse reflectance mode.
This study was undertaken to know the size and shape of the silver
nanoparticles biosynthesized using Morinda pubescens. SEM analysis RESULTS
was done using FEI Quanta 200 SEM machine. Thin films of the
sample were prepared on a carbon coated copper grid by just Biosynthesis of silver nanoparticles
dropping a very small amount of the sample on the grid, extra Plant materials were collected and plant leaf extracts were prepared
solution was removed using a blotting paper. Then the film on the both by conventional and homogenization methods. Biosynthesis of
SEM grid was allowed to dry and the images of nanoparticles were silver nanoparticles by the filtrate of Morinda pubescens was
taken. confirmed by change in the colour of the filtrate to brown after
Characterization of silver nanoparticles by Transmission addition of silver nitrate and it was optimized under various
Electron Microscopy (TEM) conditions as Sun Light, UV, Microwave, Direct Boiling, and Room
Temperature (Fig. 1a, 1b, 1c, 1d, 1e). The obtained nanoparticles
TEM is a method of producing images of a sample by illuminating the were recovered and stored (Fig. 2). This resulted due to excitation of
sample with electronic radiation (under vacuum), and detecting the surface plasmon vibrations in the silver nanoparticles.

(a) (b) (c)

(d) (e)
Fig. 1: Synthesis of silver nanoparticles under (a)Sunlight irradiation (b)UV irradiation (c)Microwave irradiation (d)Direct Boiling
(e)Room temperature.

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Asian J Pharm Clin Res, Vol 5, Suppl 1, 2012, 159-162

particle size was evident from the images. The TEM images indicated
equally spherical shaped orthorhombic crystals.

(a)

(b)
Fig. 2: Recovery of silver nanoparticles.
Fig. 4: SEM analysis of (a) silver nanoparticle in 1μm and (b)
UV-VIS spectral analysis silver nanoparticle in 5μm
The bioreduction of silver in the filtrate reaction solution was
monitored by periodic sampling of the reaction mixture at regular
intervals by using UV-Vis spectroscopy. Control flasks maintained
with silver nitrate solution (without plant filtrates) did not show any
change of colour and its absorbance maximum was found to be at
340nm, which was specific for silver nitrate solution. Sunlight
irradiated reaction mixture with plant filtrate and silver nitrate
showed a strong characteristic absorbance peak at around 430nm
(Fig. 3). Analysis by spectrophotometer was made up to 8hrs.

Fig. 5: TEM analysis of silver nanoparticle in 150nm.

XRD Analysis
XRD analysis showed distinct diffraction peaks which can be indexed
the angle vales of (111), (200), (220), (240) crystalline planes of
nano silver. This analysis revealed the orthorhombic crystals of
silver nanoparticles. The high peaks in the analysis indicated the
active silver composition with the indexing (Fig. 6).

Fig. 3: UV-Vis spectroscopy of silver nanoparticles.

CHARACTERIZATION OF SILVER NANOPARTICLES
SEM Analysis
Scanning electron microscopic analysis of the silver nitrate solution
(Control) and reduced form of silver nitrate solution were clearly
distinguishable owing to their size difference. It was clear from the
SEM pictures that control silver nitrate particles were more than
1000nm size, where as silver particles in the bioreduced colloidal
suspensions measured 15-20nm in size. Fig. 4 is the SEM of
bioreduced silver nitrate.
TEM Analysis Fig. 6: XRD pattern of the silver nanoparticles.

TEM analysis revealed that the synthesized nanoparticles were FTIR Analysis
stable in solution at room temperature. The size of nanoparticle The presence of some functional groups as revealed by FTIR spectral
ranged from 20-40nm (Fig. 5). The decrease in anisotropy and analysis is shown in Fig. 7. The FTIR Spectral analysis of silver

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nanoparticles showed certain common absorption bands at 3398 Reduction of silver ions present in the aqueous solution of silver
cm-1, characteristic of υ (O-H) and υ (N-H) vibrational frequencies. complex during the reaction with the ingredients present in the
Vibrational peaks between 2899 and 2977 cm-1 were characteristic Morinda pubescens plant leaf extract as observed by the UV-Vis
of a υ (C-H) symmetrical vibration of saturated hydrocarbon. The spectroscopy revealed the presence of silver nanoparticles . The
vibrational frequency υ (C-O) was observed in the spectra of the XRD, SEM, TEM analyses showed the particle size between 25-50nm
extracts at 1047 and 1087 cm-1. Deviation from this region to a as well the cubic structure of the nanoparticles. FTIR analysis
higher wave number was observed which was indicative of a confirmed that the bioreduction of silver ions to silver nanoparticles
secondary amide. These peaks were sharper than the υ (O-H) peaks was due to the reduction by capping material of plant extract. The
due to reduction in hydrogen bonds which increased with present study, thus showed a simple green route for rapid and
electronegativity. Vibrational peaks at 1654 cm-1 in extract signified economical synthesis of silver nanoparticles.
the possibility of an aromatic compound. Based on the physical state
of the extracts and the characteristic features of the infrared CONCLUSION
vibrational peaks in the spectra, terpenoids, long chain fatty acids The bio-reduction of aqueous silver ions by the leaf extract of the
and secondary amide derivatives were the possible compounds in Morinda pubescens plant had been demonstrated. The reduction of
the obtained nanoparticles. the metal ions through the leaf extract led to the formation of silver
nanoparticles of fairly well-defined dimensions. In the present study
it was found that leaves of Morinda pubescens could be a good source
for the synthesis of silver nanoparticles through a green chemistry
approach with several advantages.
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FTIR analysis was used further for the characterization of the
resulting nanoparticles. FTIR absorption spectra of silver
nanoparticles showed absorbance bands (before bioreduction) in
the region of 500–2000 cm-1 at 1697, 1618, 1514, 1332, 1226 cm-1.
These absorbance bands were known to be associated with the
stretching vibrations for –C C–C O, –C C– [(in-ring) aromatic], –C–C–
[(in-ring) aromatic], C–O (esters, ethers) and C–O (polyols),
respectively12. In particular, the 1226 cm-1 band arose most
probably from the C–O group of polyols such as hydroxyflavones and
catechins. The total disappearance of this band after the
bioreduction might be due to the fact that the polyols were mainly
responsible for the reduction of silver ions, whereby they
themselves got oxidized to unsaturated carbonyl groups leading to a
broad peak at 1650cm-1 for reduction of silver.

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