Indian Journal of Biotechnology

Vol 15, July 2016, pp 382-391

Utilization of the pernicious aquatic weed salvinia (Salvinia molesta D S Mitchell)

in generating gold nanoparticles
Tasneem Abbasi, J Anuradha and S A Abbasi*
Centre for Pollution Control and Environmental Engineering, Pondicherry University, Puducherry 605 014, India

Received 19 October 2014; revised 20 June 2015; accepted 18 July 2015

A biomimetic method of gold nanoparticle (AuNP) synthesis utilizing the highly invasive aquatic weed salvinia
(Salvinia molesta D S Mitchell) is presented. In an attempt to utilize the entire plant for the synthesis, the extracts of all its
parts – aerial as well as submerged – were employed in various proportions with the gold (III) solution. The formation of
AuNPs was tracked using UV-Visible spectrophotometry. The electron micrographs revealed the presence of AuNPs of
monodispersed spherical and polydispersed triangular, pentagonal and nanoflowershapes in sizes ranging 7-25 nm,
20-50 nm and 75-175 nm, respectively. The presence of gold atoms was confirmed from the EDAX and X-Ray diffraction
studies. The FTIR spectral study indicated that the proteins and polyols in the plant extract could have been responsible for
the reduction of the gold ions to gold nanoparticles and their subsequent stabilization.

Keywords: EDAX, FT-IR, gold nanoparticles, Salvinia molesta, XDR

Introduction by a simple, rapid, non-hazardous, energy saving,

Biomimetic nanoparticle (NP) synthesis, which is
based on mimicking nature’s ways to achieve energy-
efficient and eco-friendly NP production, has emerged
as a highly promising and exciting branch of
nanotechnology1. It enables the synthesis of nano
materials under ambient conditions and with little
pollution in contrast to energy intensive and/or highly
polluting chemical and physical methods. Biomimetic
routes also enable good control over the NP shapes
and sizes2,3.
Gold nanoparticles (AuNPs) are highly sought after
and find wide applications in different types of
chemical, biochemical, pharmaceutical and electronic
industries 4. Most reported work on plant-based
biological synthesis of AuNP has involved plants,
which have well-established uses as food or as
medicines, cosmetics or ornamentals5-7. Moreover, in
most such studies only one or the other part of the
plant, mainly the leaves, has been employed 2,3.
In contrast, the present study aims to utilize the
whole plant of the pernicious aquatic weed salvinia
(Salvinia molesta D S Mitchell) for the synthesis
of AuNPs. The distinctive attribute of the present
work is that it facilitates the synthesis of AuNPs
——————
*Author for correspondence:
Tel: +91-413-2654398
prof.s.a.abbasi@gmail.com
single-pot, cost-effective and environmentally safe
method. Concurrently, it enables the gainful
utilization of an obnoxious invasive aquatic weed
salvinia, which, in addition to being worthless, causes
damage to the aquatic environment and costs
enormously to eradicate8-11.
A survey of the state-of-the-art of AuNP synthesis
using plants indicates that there has been no attempt
so far to use salvinia for AuNPs synthesis. Given the
special structural, electronic, magnetic, optical,
catalytic and biocompatible properties of AuNPs, it is
imperative to develop techniques by which AuNPs of
diverse shapes and sizes can be synthesized in as
green, clean, and inexpensive manner as possible.
Hence we have explored the possible use of salvinia
in AuNP synthesis and have identified factors that can
help in directing the morphology and sizes of the
synthesized AuNPs.
Materials and Methods
Preparation of Aqueous Extracts of Aerial and Submerged
Parts of Salvinia
Salvinia was collected from its natural habitat
near Pondicherry (Central) University, Puducherry.
The fresh, mature, and disease-free plant parts (aerial
& submerged) were washed thoroughly with water,
and then dipped in saline water to sterilize their
surface, followed by washing them liberally before
 ABBASI et al: SYNTHESIS OF GOLD NANOPARTICLES BY S. MOLESTA
blotting them dry. A known quantity of plant samples
was dried at 105°C to a constant weight. On the basis
of dry wt thus obtained, extracts of plant parts (aerial
& submerged) were made by boiling 1.0 g dry wt of
the plant material with 100 mL of sterile distilled
water for 5 min. The contents were filtered through a
Whatmann no. 42 filter paper and the filtrate was
stored under refrigeration at 4°C2,3. Reconnoitery
experiments indicated that the extracts retained their
integrity upto 3 d, as evident by the extent of intensity
of nanoparticles generated by them. Hence in all the
experiments the extracts were used within 3 d of
preparation.
Au (III) solution
A 10 -3 M aqueous solution of Au (III) was prepared
using analytical reagent grade HAuCl4. The stock
solution was stored in amber bottles covered with
black sheets.
Nanoparticle Synthesis
The plant extracts (aerial & submerged) were
mixed with Au (III) solution at ambient temperature.
The AuNPs began forming almost immediately as
indicated by the appearance of pinkish red or purple
colour, which grew in intensity with time. The
developed colour and its intensity depended on the
stoichiometric ratio, in which the plant extract and the
metal ion had been mixed. The spectra of the reaction
mixtures were recorded using UV-visible
spectrophotometer. In a typical experiment the plant
extract was mixed with Au (III) solution in the
proportions as presented in Table 1.
Characterization of Synthesized AuNPs
UV-Visible Spectroscopy
UV-Visible (UV-Vis) spectroscopy is an essential
means to track the synthesis of the aqueous metal
nanoparticles. The bioreduction of the gold ions was
monitored by recording the UV-Visible spectra in the
range of 190-1100 nm for the reaction mixture filled
in quartz cuvettes with 1 cm path length. De-ionized
double distilled (in all-glass stills) water was used as
blank. The spectra were recorded by employing
Labindia (model UV 3000 +) and ELICO (model SL
164) double beam UV-Visible spectrophotometers
operated at 1 nm resolution.
SEM/TEM Studies
SEM (scanning electron microscopy) and TEM
(transmission electron microscopy) were performed to
determine the size and morphology of the synthesized
383
AuNPs. The AuNPs were centrifuged at 12,000 rpm
for 20 min using Remi C24 centrifuge. The resulting
pellets were washed thrice with de-ionized distilled
water to remove the unreacted constituents and were
re-dispersed in similar water.
The samples for SEM were prepared by placing a
drop of suspension on a carbon-coated SEM grid. For
high-resolution SEM (HR-SEM) studies, the samples
were prepared by placing dried pellets on a carbon
coated aluminium stub. The AuNPs were sampled for
TEM by first pelletizing the nanoparticles by diluting
and through sonication. The micrographs were
recorded by depositing a drop of the well-dispersed
samples on carbon coated 300 mesh placed on copper
TEM grids and excess liquid was wiped off with filter
paper.
Energy Dispersive X-Ray (EDAX) Studies
The identification of the elemental composition of
the synthesized AuNPs was performed using the
EDAX equipment attached with the SEM/HRSEM
microscopes. The EDAX spectrum was recorded
after documenting the electron micrographs in the
spot-profile mode by focusing on the densely occupied
gold nanoparticle region.
Selected Area Electron Diffraction (SAED) Studies
The SAED pattern recorder was equipped with
TEM-instrument, hence the SAED patterns were
documented along with the TEM micrographs.
X-Ray Diffraction (XRD) Studies
The powder XRD spectrum of the AuNPs was
recorded to analyse the crystallinity of the material.
An aliquot of the pelletized AuNPs was drop-casted
into thin film on a glass slide and the spectral
recording was executed by scanning in the
2θ region, from 0° to 80°, at 0.02° per min, and with
the time constant of 2 sec. The crystalline pattern of
the nanoparticles was recorded using Cu Kα1 radiation
with a wavelength (λ) of 1.5406Å at a tube voltage of
40 kV and a tube current of 30 mA.
Fourier Transform Infrared (FTIR) Spectroscopic Studies
FTIR spectroscopy was used to help identify the
functional groups involved in the reduction,
stabilization and capping of the gold nanoparticles. For
performing the FTIR study, the sample was dried
completely and ground with potassium bromide. The
spectrum was recorded at diffuse reflectance mode
with 4 cm-1 resolution in the mid-IR region between
the Wave numbers 4000 and 400 cm-1.
384 INDIAN J BIOTECHNOL, JULY 2016

Results and Discussion

UV-Visible Spectra
The UV-Visible spectrophotometric studies
confirmed that AuNPs were formed on mixing the
plant extract with Au (III) solution at ambient
conditions of temperature and pressure. The reaction
mixtures exhibited pinkish red/violet colour within 10
min of the start of the reaction. It gave rise to the
surface plasmon resonance (SPR) absorbance band
attributable to the combined vibration of free
conducting electrons of AuNPs in resonance with the
electromagnetic field12.
Fig. 2—UV-Visible spectra of AE 2 gold nanoparticles synthesis.
SPR spectral bands obtained from the tracking of
the AuNPs synthesis varied depending on the plant
part used for making the extract and its proportions
with respect to the Au(III) solution in the reaction
medium (aerial: AE1 & AE 2; submerged: SE1 & SE2;
Table 1). The resulting spectra showed the presence
of two types of spectral patterns: one with a single
λmax in the visible region, and the other with two
λmax―one in the visible region and the other in the
NIR (near infra-red) region. The reaction components
of AE1 and SE1 (Figs 1, 3a, 4 & 6a) revealed the
presence of single peak, while the AE2 and SE2
(Figs 2, 3b, 5 & 6b) combinations showed the
presence of two peaks in their spectra. The presence
Table 1—Extract and metal solution combinations studied for
gold nanoparticles synthesis using S. molesta
Conc. of component in the
Plant part used for reaction mixture
Plant used (mg/ L)
preparing extract
Extract Au(III)
Aerial (AE 1) 1000 30
Aerial (AE 2) 7000 100
S. molesta
Submerged (SE 1) 1000 30
Submerged (SE 2) 1000 17 Fig. 3 (a & b)—Plot of intensity of surface plasmon resonances
(SPRs) of (a) AE1 and (b) AE 2 against the reaction duration.

Fig. 1—UV-Visible spectra of AE1 gold nanoparticles synthesis. Fig. 4—UV-Visible spectra of SE 1 gold nanoparticles synthesis.
 ABBASI et al: SYNTHESIS OF GOLD NANOPARTICLES BY S. MOLESTA 385

Table 2—Summary of results obtained from the UV-Visible

spectral studies for gold nanoparticles synthesized
using S. molesta extracts
Observation Spectra Extract+metal ion mixture
after observed
AE 1 AE 2 SE1 SE 2
(h)
0 λmax - - 531 -
Abs. 0.055
2 λmax 527 544 1087 543 580
Abs. 0.333 1.735 1.421 0.200 0.152
4 λmax 527 541 1099 543 580
Abs. 0.368 1.953 1.519 0.244 0.199
6h λmax 527 538 1082 540 577
Fig. 5—UV-Visible spectra of SE 2 gold nanoparticles synthesis.
Abs. 0.380 2.051 1.663 0.281 0.223
24 λmax 525 537 1090 531 577
Abs. 0.413 2.025 1.648 0.334 0.299
48 λmax 522 539 1087 531 566
Abs. 0.447 1.882 1.418 0.345 0.226

the synthesized nanoparticles are with an intrinsic

Fig. 6—Plot of intensity of surface plasmon resonances (SPRs) of
(a) SE 1 and (b) SE2 against the reaction duration.
of a single prominent peak in the visible region of
AE1 and SE1 spectra indicates that it might have
occurred due to the transverse plasmon resonance
(TPR), which is exhibited by spherical nanoparticles.
This observation was confirmed by the presence of
spherical particles in the electron micrographs. On the
other hand, the spectra of the AE2 reaction
combination had two clear absorption bands, a shorter
wavelength transverse absorption band in the visible
region (out of plane vibration band) and a longer
wavelength longitudinal absorption band in the NIR
region (in plane plasmon vibrations), illustrating that
anisotropy13-15. As the SPR (surface plasmon
resonance) band measured in SE2 extends from the
visible to the NIR region, the particles formed can be
regarded as complex hybrids of spheres and stars that
are not highly monodisperse and their individual
longitudinal plasmon resonance bands could have
been lost in ensemble measurements16. This was also
later confirmed from the SEM and TEM studies.
The results obtained from the spectrophotometric
studies for all the reaction combinations are
summarized in Table 2. It illustrates that the intensity
of the nanoparticles formation was found to increase
gradually with an increase in reaction duration, upto a
point. The rate of reduction of gold ions present in the
reaction medium was found to be more than 90%
complete within 6 h (AE2) and 24 h (AE1, SE1 & SE2)
of the reaction duration.
AuNPs generated from all the reaction
combinations were characterized for their size,
morphology, purity and crystal structure by
employing SEM, HR-SEM, TEM, EDAX, XRD and
SAED techniques. The FTIR spectroscopy was used
for the identification of biomolecules that could be
possibly involved in the bioreduction of the gold ions
into AuNPs and the consequent stabilization and
capping of the synthesized AuNPs.
Electron Microscopic (SEM, HR-SEM, TEM) and EDAX
studies
The electron micrographs (SEM & HR-SEM) of
AuNPs generated from the AE1 and SE1 reaction
combinations (Figs 7 & 9) had particles of a spherical
386 INDIAN J BIOTECHNOL, JULY 2016

Fig. 7 (a-c)—A composite visual of (a) scanning electron micrograph (inset is EDX spectrum); (b) & (c) high resolution scanning
electron micrographs of AE1 gold nanoparticles

Fig. 8 (a-c)—A composite visual of (a) scanning electron micrograph (inset is EDX spectrum); (b) & (c) high resolution scanning
electron micrographs of AE2 gold nanoparticles.

Fig. 9 (a-c)—A composite visual of (a) scanning electron micrograph (inset is EDX spectrum); (b) & (c) high resolution scanning
electron micrographs of SE1 gold nanoparticles.
shape. TEM images revealed that the size of
the synthesized particles were in the range of
16.8-23.6 nm and 7-25 nm in AE1 and SE1 reaction
combinations, respectively (Figs 11 & 13).
Figs 8, 10, 12 and 14 present the SEM, HR-SEM
and TEM micrographs of biogenic AuNPs obtained
from AE2 and SE2 reaction mixtures. The micrographs
reveal that the generated particles were anisotropic
with triangular, pentagonal and nanoflower shapes.
The sizes of the nanoparticles synthesized from AE2
and SE2 reaction combinations were observed to be in
the ranges from 20-50 nm and 75.5-175.8 nm,
respectively.
The EDAX spectrum recorded in spot-profile mode
confirmed the presence of gold atoms by exhibiting
strong signals from the AuNPs (insets of Figs 7-10).
 ABBASI et al: SYNTHESIS OF GOLD NANOPARTICLES BY S. MOLESTA 387

Fig. 10 (a-c)—A composite visual of (a) scanning electron micrograph (inset is EDX spectrum); (b) & (c) high resolution scanning
electron micrographs of SE2 gold nanoparticles.

Fig. 11 (a-f)—A composite visual of transmission electron Fig. 13 (a-f)—A composite visual of transmission electron
micrographs: (a-e) Showing spherical particles of AE 1 at different micrographs: (a-e) Showing spherical particles of SE1 at different
magnifications, & (f) SAED pattern of the gold nanoparticles. magnifications, & (f) SAED pattern of the gold nanoparticles.

Fig. 12 (a-f)—A composite visual of transmission electron Fig. 14 (a-f)—A composite visual of transmission electron
micrographs: (a-e) Showing spherical and anisotropic particles of micrographs: (a-e) Showing anisotropic nanoflowers of SE 2 at
AE2 at different magnifications, & (f) SAED pattern of the gold different magnifications, & (f) SAED pattern of the gold
nanoparticles. nanoparticles.

Weak signals were also witnessed from C, N and An optical absorption band characteristic of AuNPs
O atoms, which can be attributed to the X-ray was prominently seen at approx 2 keV position17,18.
emission from the biomolecules of residual plant Figs 11f-14f represent the SAED patterns
extracts present along with the synthesized AuNPs. recorded from the AuNPs. The presence of bright
388 INDIAN J BIOTECHNOL, JULY 2016

circular spots was in agreement with the Bragg’s obtaining the FWHM of the (111) Bragg’s reflection
planes representing the crystalline nature of the from the X-ray diffracto grams21.
synthesized AuNPs19. The electron micrographs The synthesized AuNPs were found to have
(Figs 11-14) show that the nanoparticles were stable average crystallite sizes of 13.2 to 19.3 nm.
and no agglomeration of the particles occurred in the The particle size as observed from XRD pattern
solution after centrifugation. (Figs 15a & c) are in close agreement with the
average size ca. 17.3 nm, indicated by the electron
XRD Studies micrographs. This denotes the formation of mono
The powder XRD patterns of AuNPs are shown in dispersed spherical particles. The particle size
Fig. 15. The diffraction spectra show intense peaks at calculated from XRD pattern (Figs 15b & d) was
2θ position that are characteristic of face-centered lesser compared to that of the size determined from
cubic structure of the AuNPs, corresponding to the the electron micrographs. This was perhaps due to the
(111), (200), (220) and (311) Bragg's planes20
Table 3—2θ position of the Bragg’s plane observed from the
(Table 3). The XRD patterns of the AuNPs, which X-ray diffractograms
match with the database of JCPDS file no. 04-0784,
Bragg’s plane (111) (200) (220) (311)
denote that the synthesized AuNPs are of pure
crystalline nature. Bragg’s peak widens with the AE1 38.11 44.31 64.45 77.37
2θ position

progress in nanoparticles formation. The crystalline AE2 38.87 44.74 64.87 78.17
domain size of the synthesized nanoparticles were SE1 38.21 44.57 64.87 78.33
calculated using the Debye-Scherrer's equation by SE2 38.79 44.75 65.05 78.17

Fig. 15 (a-d)—X-ray diffraction (XRD) spectrum of (a) AE 1, (b) AE 2, (c) SE1, & (d) SE2 (*peaks due the biomolecules found in the
vicinity of gold nanoparticles).
 ABBASI et al: SYNTHESIS OF GOLD NANOPARTICLES BY S. MOLESTA 389

polycrystalline nature of the synthesized gold
nanoparticle22. The ratio of intensity between the
(200) and (111) Bragg’s diffraction peaks are in the
range of 0.20-0.24. This is found to be lesser than
the intensity ratio (0.52) of conventional bulk gold,
confirming the presence of nanoparticles with (111)
facets
FTIR Spectroscopy
FTIR spectral studies were employed to identify
the biomolecules present in the plant extract, which
could have interacted with the gold ion solution and
would have been responsible for the bioreduction and
capping/stabilization of the AuNPs. The spectrum
(Fig. 16) indicates that proteins and polyols of
terpenoids, flavonoids and polysaccharides were
probably responsible for the bioreduction and
capping/stabilization of the synthesized nanoparticles.
The FTIR pattern of aerial extract showed the
presence of medium or strong absorption bands at
1662.9, 1541.8, 1314.4 and 1073 cm−1 in the region
1000-1800 cm−1. The band at 1662 cm-1 can be
assigned to C=O stretch of the amide I band23,24. The
bands at 1541 cm-1 and 1314 cm-1 correspond to
amide I and III bands of polypeptides/proteins. The
band at 1073.0 cm−1may be due to the C–N stretching

Fig. 16—FTIR spectrum of S. molesta extract and the synthesized gold nanoparticles.
390 INDIAN J BIOTECHNOL, JULY 2016
corresponding to amine group. The spectrum of
AuNPs showed the presence of absorbance bands
centered at 1910.7, 1738.1, 1631.6, 1525.1, 1275.6
and 1011.8 cm−1. The bands at 1738 and 1631 cm−1
can be attributed to polyols, such as, flavonoids,
terpenoids and polysaccharides. The bands at 1525,
1275 and 1011 cm−1 can be assigned to vibrations of
primary and secondary amines, polyols, etc. The
appearance of new bands at 1910 cm−1and 1738 cm−1
and shift in bands at 1662, 1541, 1314 and 1073 cm−1
can be attributed to the binding of amides and polyols.
This shows that amines and polyols found in the aerial
part of the plant could have been responsible for the
reduction of the gold ions atoms and also their
efficient capping when the atoms aggregated to
nanosizes, thereby ensuring their stability.
The FTIR spectrum of extract prepared from the
submerged part of the weed (Fig. 16) shows
absorbance bands centered at 1729.5, 1608.6, 1547.1,
1388.1 and 1110.8 cm−1. The spectrum after
bioreduction shows some absorbance bands centered
at 1632.2, 1383.1 and 1021.6 cm−1. The bands at
1632.2 and 1383.1 cm−1 can be attributed to polyols,
such as, flavones, terpenoids and polysaccharides 24.
The band at 1021.6 cm−1 can be attributed to aliphatic
amines or alcohols/phenols. The shift in bands at
1729.5, 1608.6, 1547.1, 1368.4 and 1110.8 cm−1 and
appearance of new bands at 1632.2, 1383.1 and
1021.6 cm−1 are likely due to the amides, sulfonyl
groups and polyols, which might have been involved
in the AuNPs formation. It is known that proteins can
bind to gold nanoparticles either through free amine
groups or cysteine residues in them23,24. Hence it can
be inferred that proteins and polyols of terpenoids,
flavonoids and polysaccharides could have been
involved in the bioreduction and capping/stabilization
of the gold nanoparticles.
Conclusion
Extracts of the aerial and the submerged parts of
salvinia (S. molesta) have been utilized in the
synthesis of gold nanoparticles (AuNPs). Based on
the plant part used for making the extract and its
proportion with reference to the gold (III) solution,
AuNPs of different sizes and morphology were
generated, ranging from isotropic spherical to
anisotropic triangular, pentagonal and flower shaped
particles of different sizes. The electron micrographs
revealed that the spherical particles were of size
ranging from 7 to 25 nm, and anisotropic particles had
sizes ranging from 20 to 175.8 nm. The synthesized
particles were shown to be pure and crystalline by
EDAX, XRD and SAED studies. FTIR spectral
studies indicated that proteins and polyols of
terpenoids, flavones and polysaccharides were
involved in the bioreduction and capping/stabilization
of gold ions into AuNPs.
Acknowledgement
SAA thanks the University Grant Commission,
Government of India, New Delhi for support in the
form of Emeritus Fellowship.
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