D. Ramya

K. Manivannan

P. Anantharaman

R. Arumugam

R. Ragupathi Raja Kannan

Applied Nanoscience

Springer - Publisher Connector

Green synthesis of silver nanoparticles using marine macroalga Chaetomorpha linum

ORIGINAL ARTICLE
Green synthesis of silver nanoparticles using marine macroalga
Chaetomorpha linum
R. Ragupathi Raja Kannan • R. Arumugam •
D. Ramya • K. Manivannan • P. Anantharaman
Received: 7 March 2012 / Accepted: 17 April 2012 / Published online: 18 May 2012
 The Author(s) 2012. This article is published with open access at Springerlink.com
Abstract The present investigation demonstrates the
formation of silver nanoparticles by the reduction of the
aqueous silver metal ions during exposure to the seaweed
(Chaetomorpha linum) extract. The silver nanoparticles
obtained were characterized by UV–visible spectrum,
FTIR and scanning electron microscopy. The characteristic
absorption peak at 422 nm in UV–vis spectrum confirmed
the formation of silver nanoparticles. The colour intensity
at 422 nm increased with duration of incubation. The size
of nanoparticles synthesized varied from 3 to 44 nm with
average of *30 nm. The FTIR spectrum of C. linum
extract showed peaks at 1,020, 1,112, 1,325, 1,512, 1,535,
1,610, 1,725, 1,862, 2,924, 3,330 cm-1. The vibrational
bands corresponding to the bonds such as –C=C (ring),
–C–O, –C–O–C and C=C (chain) are derived from water-
soluble compounds such as amines, peptides, flavonoids
and terpenoids present in C. linum extract. Hence, it may
be inferred that these biomolecules are responsible for
capping and efficient stabilization. Since no synthetic
reagents were used in this investigation, it is environmen-
tally safe and have potential for application in biomedicine
and agriculture.
Keywords Marine algae  Green chemistry 
Silver nanoparticles  Flavonoids
Introduction
Recently, silver nanoparticles (AgNPs) are widely investi-
gated to owing their broad range of applications as antibac-
terials (Venkatpurwar and Pokharkar 2011; Wang Yh et al.
2008), biosensors (Chen et al. 2007) and in plant growth
metabolism (Krishnaraj et al. 2012). The green synthesis of
inherently safer AgNPs depends on the adoption of the basic
requirements of green chemistry: the solvent medium, the
benign reducing agent and the non-hazardous stabilizing
agent (Vigneshwaran et al. 2006). Compared to microbe-
assisted synthesis, plant-mediated synthesis of nanoparticles
is a relatively underexploited field and is recently gaining
wide attention. AgNPs have been successfully synthesized
using several plant extracts (Shankar et al. 2003; Amkamwar
et al. 2005; Chandran et al. 2006; Li et al. 2007; Venkatpurwar
and Pokharkar 2011). All these investigations are restricted to
terrestrial plants, but only limited reports are available for
synthesis of nanoparticles from marine plants (Govindaraju
et al. 2009; Nabikhan et al. 2010; Venkatpurwar and Po-
kharkar 2011). Marine macroalgae produce a great variety of
secondary metabolites that showed therapeutic potential
(Tierney et al. 2010; Mayer et al. 2011). Chaetomorpha linum
(Muller) Kutzing is a green seaweed mainly acknowledged
for its ecological role as possible regulator of nutrient avail-
ability in estuarine habitats (Krause-Jensen et al. 1999). Many
researchers (Heo et al. 2005; Patra et al. 2009; Prabhahar et al.
2011) have reported that C. linum exhibits medicinal prop-
erties with high-value phytochemicals, which motivated us to
carry out the present investigation on the synthesis of AgNPs
using C. linum biomass.
R. R. R. Kannan (&)  R. Arumugam  D. Ramya 
K. Manivannan  P. Anantharaman (&)
Faculty of Marine Sciences, CAS in Marine Biology, Annamalai
University, Parangipettai, 608 502 Tamil Nadu, India
e-mail: panantharaman@gmail.com
Present Address:
R. R. R. Kannan
Research Centre for Plant Growth and Development, School
of Life Sciences, University of KwaZulu-Natal Pietermaritzburg,
Private Bag X 01, Scottsville 3209, South Africa
123
Appl Nanosci (2013) 3:229–233
DOI 10.1007/s13204-012-0125-5
Materials and methods
Chemicals
Silver nitrate (AgNO3) was purchased from Merck, India.
All other reagents used in this present investigation are of
analytical grade.
Sample collection and preparation
The green seaweed C. linum (Muller) Kutzing was col-
lected from the intertidal region of Kanyakumari coast
located at southeast coast of India. Collected sample was
immediately brought to the laboratory in new plastic bags
containing natural sea water to prevent evaporation. Plants
were washed thoroughly with tap water to remove extra-
neous materials and shade-dried for 5 days and oven-dried
at 60 C until constant weight was obtained, then ground
into fine powder using electric mixer and stored at 4 C for
future use.
Preparation of seaweed extract
Dried finely powdered C. linum (10 g) was boiled with
100 ml of distilled water at 60 C for 15 min. The extract
was filtered through nylon mesh, followed by Millipore
filter (0.45 lm) and used for further experiments.
Synthesis of silver nanoparticles
The Erlenmeyer flask containing 100 ml of AgNO3
(1 mM) was reacted with 20 ml of the aqueous extract of
C. linum. This setup was incubated in dark (to minimize the
photoactivation of silver nitrate) at 37 C under static
condition. A control setup was also maintained without C.
linum extract.
Characterization of silver nanoparticles
The bioreduction of AgNO3 was confirmed by sampling
the reaction mixture at regular intervals and the absorption
maximum was scanned by UV–vis spectra, at the wave-
length of 300–700 nm in Perkin Elmer Lamda 25 spec-
trophotometer. Further, the bioreduced reaction mixture
was subjected to centrifugation at 75,0009g for 30 min,
resulting pellet was dissolved in de-ionized water and fil-
tered through Millipore filter (0.45 lm). An aliquot of this
filtrate containing AgNPs was used for FTIR and SEM
studies. For electron microscopic studies, 25 ll of the
sample was sputter-coated on copper stub and the images
of nanoparticles were studied using SEM (JEOL, Model
JFC-1600).
Results and discussion
Reduction of AgNO3 was visually evident from the colour
change (brownish-yellow) of reaction mixture after 30 min
of reaction (Fig. 1). Intensity of brown colour increased in
direct proportion to the incubation period. It may be due to
the excitation of surface plasmon resonance (SPR) effect
and reduction of AgNO3 (Mulvaney 1996). The control
AgNO3 solution (without seaweed extract) showed no
colour change. Absorption spectrum of reaction mixture at
different wavelengths ranging from 400 to 700 nm
revealed a peak at 420 nm (Fig. 2). The characteristic
absorption peak at 422 nm in UV–vis spectrum (Figs. 2, 3)
confirmed the formation of AgNPs. In the present study the
Fig. 1 Colour intensity of the
Chaetomorpha linum extract
incubated with silver ions at the
beginning of reaction and after
48 h reaction
230 Appl Nanosci (2013) 3:229–233
123
colour intensity at 422 nm increased with duration of
incubation. This is similar to the surface plasmon vibra-
tions with characteristic peaks of silver nanoparticles pre-
pared by chemical reduction (Petit et al. 1993; Kong and
Jang 2006).
A scanning electron microscope was employed to analyse
the structure of the nanoparticles that were formed. From
Fig. 4 it is evident that the AgNPs coalesced to nano-clus-
ters (white marking). When reaction mixtures were incu-
bated for 48 h some nanoparticles aggregated. The size of
nanoparticles synthesized in the present study by C. linum
biomass varied from 3 to 44 nm with average of *30 nm.
Jain et al. (2009) reported particle size between 25 and
50 nm in cubic structure synthesized by papaya fruit extract
and Cataleya (2006) reported macrophages in comparison to
larger nanoparticles (i.e. 30–55 nm).
FTIR measurements were carried out to identify the
possible biomolecules responsible for the stabilization of
the newly synthesized AgNPs. Figure 5 represents the FTIR
spectrum of C. linum extract shows peaks at 1,020, 1,112,
1,325, 1,512, 1,535, 1,610, 1,725, 1,862, 2,924, 3,330 cm-1.
The FTIR spectra of dried C. linum extract showed strong
absorption band at 1,535 cm-1 corresponding to bending
vibration of secondary amine of proteins. Another band
observed at 1,610 cm-1 is assigned to the stretching
vibration of (NH) C=O group. After reduction of AgNO3
the decreases in intensity at 1,535 cm-1 signify the
involvement of the secondary amines in the reduction pro-
cess. On the other hand, the shift of band from 1,610 cm-1
is attributed to the binding of (NH) C=O group with
nanoparticles. Since a member of (NH) C=O group within
the cage of cyclic peptides is involved in stabilizing the
nanoparticles, the shift of (NH) C=O band is quite small.
Thus, the peptides play a major role for the reduction of Ag*
to Ag nanoparticles. Bands at 1,020 cm-1 can be assigned
as absorption peaks of –C–O–C. The bands at 1,325 cm-1
in silver nano may be attributed to –C–O stretching mode
(Shankar et al. 2003; Huang et al. 2007; Philip 2009a, b).
Strong IR band at 1,512 cm-1 in the spectrum of silver
nanoparticle arise from the stretching vibrations of C=C
Fig. 2 UV–visible spectrum of
C. linum extract treated with
and without AgNPs during
different wavelengths
Fig. 3 Colour intensity at
422 nm for AgNPs synthesis by
C. linum during different time
intervals
Appl Nanosci (2013) 3:229–233 231
123
chain (Schulz and Baranska 2007). The characteristic peak
at 1,725 cm-1 was noticed in the C. linum extract and this
may be due to the free amine groups which are used for the
stabilization of silver nanoparticles (Philip 2010). The
vibrational bands corresponding to the bonds such as –C=C
(ring), –C–O, –C–O–C and C=C (chain) are derived from
water-soluble compounds such as flavonoids and terpenoids
present in C. linum extract. Hence, it may be inferred that
these biomolecules are responsible for capping and efficient
stabilization.
Conclusion
In this present investigation, we described environment-
friendly synthesis of AgNPs using marine macroalgae
Fig. 4 SEM image of AgNPs
synthesized by C. linum
Fig. 5 FTIR spectrum of AgNPs synthesized by C. linum extract
232 Appl Nanosci (2013) 3:229–233
123
C. linum. The amines, peptide groups and secondary
metabolites flavonoids and terpenoids present in the
C. linum extract were involved in the bioreduction and
stabilization of AgNP. This method of AgNPs synthesis
does not use any toxic reagents and thus has the potential
for use in biomedical and agricultural applications.
Acknowledgments The authors are grateful to Prof. T. Balasubra-
manian, Dean, Centre of Advanced Study in Marine Biology, Faculty
of Marine Sciences and authorities of Annamalai University for
providing the necessary facilities.
Open Access This article is distributed under the terms of the
Creative Commons Attribution License which permits any use, dis-
tribution, and reproduction in any medium, provided the original
author(s) and the source are credited.
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Green synthesis of silver nanoparticles using marine macroalga Chaetomorpha linum

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