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Colloids and Surfaces B: Biointerfaces 87 (2011) 159–163

Contents lists available at ScienceDirect

Colloids and Surfaces B: Biointerfaces

journal homepage: www.elsevier.com/locate/colsurfb

Facile green synthesis of gold nanoparticles using leaf extract of antidiabetic

potent Cassia auriculata
V. Ganesh Kumar a,∗ , S. Dinesh Gokavarapu b , A. Rajeswari b , T. Stalin Dhas a , V. Karthick a ,
Zainab Kapadia b , Tripti Shrestha b , I.A. Barathy b , Anindita Roy b , Sweta Sinha b
a
Nanoscience Division, Centre for Ocean Research, Sathyabama University, Chennai 600 119, India
b
Department of Biotechnology, Sathyabama University, Chennai 600 119, India

a r t i c l e i n f o a b s t r a c t

Article history: A simple biological method for the synthesis of gold nanoparticles (AuNPs) using Cassia auriculata aqueous
Received 8 February 2011 leaf extract has been carried out in the present study. The reduction of auric chloride led to the formation
Received in revised form 5 May 2011 of AuNPs within 10 min at room temperature (28 ◦ C), suggesting a higher reaction rate than chemical
Accepted 8 May 2011
methods involved in the synthesis. The size, shape and elemental analysis were carried out using X-ray
Available online 13 May 2011
diffraction, TEM, SEM-EDAX, FT-IR and visible absorption spectroscopy. Stable, triangular and spherical
crystalline AuNPs with well-defined dimensions of average size of 15–25 nm were synthesized using C.
Keywords:
auriculata. Effect of pH was also studied to check the stability of AuNPs. The main aim of the investigation
Biosynthesis
Gold nanoparticles
is to synthesize AuNPs using antidiabetic potent medicinal plant. The stabilizing and reducing molecules
Cassia auriculata of nanoparticles may promote anti-hyperglycemic if tested further.
Leaf extract © 2011 Elsevier B.V. All rights reserved.
Electron microscopy

1. Introduction ties. Based on the capping agents, environmentally benign AuNPs

Gold nanoparticles (AuNPs) are very important nanoscale mate-
rials that have been studied extensively as they exhibit completely
new and improved properties when compared to the bulk met-
als. Novel methods involved in the preparation of nanoparticles of
noble metals like gold has gained importance due to its remarkable
size-dependant optical and electronic properties. Gold nanoparti-
cles are unique due to their tunable Surface Plasmon Resonance
(SPR); hence they are used in bio-labeling [1], biosensors devices
for the detection of viruses and bacteria [2], biomedical sci-
ence including drug delivery [3], tissue/tumor imaging, photo
thermal therapy and immuno-chromatographic identification of
pathogens in clinical specimens [4]. Due to the recent awareness
of green chemistry, it has been concluded that gold nanoparti-
cles are biocompatible, inert [5], bind readily to a large range
of biomolecules such as proteins/enzymes [6,7], DNA [8], amino
acids [9,10] and expose large surface area for the immobilization
ion of such biomolecules and environmentally friendly chemicals.
Biosynthesis of metal nanoparticles using natural products is an
emerging area of nanoscience research. Natural environment is a
rich source of crude untreated extracts of plants, marine organisms
and microorganisms and it is crucial to investigate their proper-
∗ Corresponding author. Tel.: +91 44 24500646.
E-mail address: ganesv@gmail.com (V. Ganesh Kumar).
using biological components will have excellent effect in biomedi-
cal applications.
There are several reports on the use of natural material sources
like plants [11,12] and algae [13,14] in the synthesis of gold
nanoparticles. The microorganisms such as bacteria [15], fungi
[16,17] and yeast [18] have been exploited for toxic metals recovery
via reduction of the metal ions. The integration of green chem-
istry principles to nanotechnology is one of the key issues in
nanoscience research. In continuation of the efforts for synthesiz-
ing gold nanoparticles by green route, a facile, rapid and single-pot
aqueous biosynthesis of gold nanoparticles using the leaf extract
of Cassia auriculata (Tanners cassia) has been reported here. C.
auriculata, a sacred shrub of India, is well-known for its medicinal
use which is used for the treatment of diabetes, urinary disorders,
ulcers, skin diseases and chronic fever [19]; its anti-hyperglycemic
and anti-hyperlipidemic properties are also well-known [20].
The important ingredients of the extract are flavoniods, tannins,
steroids, carbohydrates and amino acids. Functionalized AuNPs
have applications in various fields such as biomedical, catalysis
and DNA detection [21–23]. Application of AuNPs in these fields
is dependent on the functionalization to synthesize nanoparti-
cles with desired shape, size and monodispersity. Functionalized
nanoparticles should be stable without undergoing degradation or
partial oxidation. There are several physical and chemical methods
for the synthesis of metallic nanoparticles that are followed by the
material scientists currently [24] but the importance of the stabi-

0927-7765/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.colsurfb.2011.05.016
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160 V. Ganesh Kumar et al. / Colloids and Surfaces B: Biointerfaces 87 (2011) 159–163

Fig. 1. (A) Auric chloride, (B) C. auriculata extract, (C) ruby red color indicating the formation of gold nanoparticles. (For interpretation of the references to color in this figure
legend, the reader is referred to the web version of the article.)

lizing biomolecules in the green synthesis and its stability has not of the respective solutions drop coated onto glass substrate on a
been studied intensively. In the present investigation, the synthesis Rich Seifert P3000 instrument operated at a voltage of 40 kV and
of gold nanoparticles emphasizes the potentiality of C. auriculata in a current of 30 mA with Cu K␣1 radiations. FT-IR of Perkin Elmer
reducing and stabilizing gold nanoparticles. The gold nanoparticles spectrophotometer was used to identify the possible biomass at
synthesized were characterized using UV–vis spectroscopy, FT-IR, a resolution of 4 cm−1 in the range of 4000–450 cm−1 and the FT-
XRD, SEM-EDAX and TEM analysis. The synthesis was carried out IR spectrum was recorded by employing KBr pellet technique using
at room temperature (28 ◦ C) and the nanoparticles formed were Perkin Elmer model-983/G detector double beam spectrophotome-
around 15–25 nm in size. ter. TEM studies were prepared by drop coating Au nanoparticles
onto carbon-coated TEM grids using Philips Technai-10. The film on
the TEM grids were allowed to dry, further the extra solution was
2. Materials and methods
removed using a blotting paper. SEM-EDAX was studied to check
the chemical composition atoms count and the surface morphology
2.1. Materials
of the AuNPs formed using XL30 FSEM, Philips.
Chloroauric acid (HAuCl4 ·3H2 O) was obtained from Loba
Chemie, India and used as received. All other reagents used in the 3. Results
reaction were of analytical grade with maximum purity. C. auricu-
lata leaves were collected from forest area of Vellore, Tamilnadu, UV–vis spectroscopy was ascertained to check the formation
India, and was cleaned with double distilled water and shade-dried and stability of AuNPs in aqueous solution. The scale of wavelength
for a week at room temperature and further C. auriculata leaves was fixed between 200 and 800 nm, the SPR of the gold nanopar-
were ground to powder and stored for further study. ticles formed corresponded to 536 nm and there was an increase
in intensity till 10 min as a function of time without any shift in
the peak wavelength (Fig. 2.) [25]. The interaction of nanoparti-
2.2. Synthesis and characterization of gold nanoparticles
cles with biomolecules of C. auriculata showed intense peaks at
2884, 1600,1507,1387,1074 and 1335 cm−1 relative shift in posi-
C. auriculata leaf extract was prepared by taking 3 g of dry leaf
tion and intensity distribution were confirmed with FT-IR (Fig. 3A
powder with 30 mL of distilled water in a conical flask along with
3 mL of methanol (minimum methanol was added in order to initi-
ate the isolation of compounds). The extract was placed in orbital
shaker for 1 h and the extract was filtered. For the synthesis of gold
nanoparticles various concentrations of leaf extracts were tried and
then the extract to be used was optimized to 1 mL. Further, 1 mL of
the extract was added to 10 mL of 1 mM auric chloride solution
and the solution was placed in orbital shaker at room temperature,
for reduction of Au3+ to Au0 . Fig. 1(A–C) shows the color change
involved in the formation of gold nanoparticles. The reaction was
rapid as the ruby red color appeared within 10 min and appear-
ance of ruby red color in the reaction confirmed the formation of
gold nanoparticles and there was no color change further. The light
absorption pattern of the plant biomass was kinetically monitored
and was in the range of 536 nm recorded using Varian-Cary 100
UV–vis spectrophotometer.
The experiment was duplicated thrice to confirm the reduc-
tion reaction. C. auriculata reduced AuNPs were centrifuged at
10,000 rpm for 15 min to prepare pellet, and it was washed with
deionized water to remove the remaining biomass. XRD measure- Fig. 2. UV–vis spectrum recorded as a function of time of reaction of aqueous solu-
ment of the C. auriculata reduced AuNPs was carried out on films tion of chloroauric acid with C. auriculata extract.
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V. Ganesh Kumar et al. / Colloids and Surfaces B: Biointerfaces 87 (2011) 159–163 161

and B) recorded for dry powder of C. auriculata, where the strong

bands were observed at 2832,1731,1612 and 1403 cm−1 . Compar-
ing both FT-IR spectra it can be identified that the changes in the
–COOH group for –OH, i.e., hydroxyl group the peak appeared at
3316 cm−1 in raw material, but after encapsulation of nanoparti-
cles, the peak is narrower and shifted to 3635 cm−1 and also for –C–
of carboxylic group the peak intensity reduced after encapsulation
of nanoparticles. The band appearing at 1387 cm−1 corresponds to
C–N stretching of amine group [26] and in the raw extract the peak
was broad and blend, but after encapsulation of nanoparticles the
peak was narrow and sharper. This implies that –COOH group in
the compound is attached to the gold nanoparticles and there in a
clear change in the spectra. 1731 cm−1 in –C– bond stretching after
the encapsulation this stretching is masked or disappeared.
The morphology and size of the particles is determined by
the TEM images and they are shown in Fig. 4(A–D). The parti-
cles formed were spherical, hexagonal and triangular in shape. The
nanotriangles formed where shown to have high surface area. The
nanoparticles formed were in the range of 15–25 nm in size, ca.
of 17 nm. The particles were monodisperse, with only a few parti-
cles of different size. The crystalline nature of AuNPs was carried
out using XRD where three diffraction peaks were observed in the
2 range of 30–80◦ , which can be indexed as (1 1 1),(2 0 0),(2 2 0)
reflections of fcc structure metallic gold respectively similar to Joint
Committee on Powder Diffraction Standards(JCPDS) file no:04-
0784 revealing that synthesized AuNPs are of pure crystalline gold.
The XRD patterns in (Fig. 5) of AuNPs obtained were similar to the
results reported earlier [26] The particle size of the AuNPs formed
were calculated using Debye–Scherrer equation which was around Fig. 3. FT-IR spectra of (A) C. auriculata extract, (B) gold nanoparticles synthesized
17 nm, were good in agreement with TEM results also. SEM micro- by C. auriculata extract.

Fig. 4. TEM images indicating the presence of hexagonal, spherical and triangular nanoparticles recorded at various magnifications (A–D).
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162 V. Ganesh Kumar et al. / Colloids and Surfaces B: Biointerfaces 87 (2011) 159–163
Fig. 5. XRD pattern of gold nanoparticles obtained using C. auriculata.
graphs show aggregates of gold nanoparticles and the particles are
in the range of 15–25 nm and there are not in direct contact even
within the aggregates indicating the stabilization of nanoparticles
by capping agents (Fig. 6A). In EDAX strong signals were observed
from the gold atoms in the nanoparticles and weaker signals for
carbon, oxygen and chloride were provenients from biomolecules
of C. auriculata (Fig. 6B).
Fig. 6. (A) SEM and (B) EDAX images showing the presence of gold nanoparticles
and bioorganic components of C. auriculata.
The pH of AuNPs synthesized using C. auriculata was 3.4 and
these nanoparticles were stable with no major shift when sodium
hydroxide of 0.1 M was added to it. The SPR was studied before and
after the addition of sodium hydroxide (base) to the nanoparticles.
The change in pH does not affect the stability of the AuNPs in the
range of 3.4–10.2.
4. Discussion
Concerning the synthesis of nanoparticles using biological com-
ponents, the potential applications of biosynthesized nanoparticles
as new advanced nanomaterials are currently studied [27]. In this
view, the locally available plant with highest antidiabetic potential
C. auriculata [28] has been selected in the study to synthesize AuNPs
in simple and eco-friendly method. During the synthesis increase
in concentration of C. auriculata leaf extract led to aggregation of
gold nanoparticles, so the condition was maintained such that 1 mL
was added to 10 mL of 1 mM auric chloride during synthesis. It is
well-known that AuNPs exhibit ruby red color in aqueous solu-
tion due to excitation of surface plasmon vibrations in AuNPs [29].
Therefore the reduction of AuCl4 − ions during exposure to the C.
auriculata extract may be easily followed by UV–vis spectroscopy.
The appearance of the ruby red color and the maximum absorbance
were indications of the formation of colloidal gold particles in
the medium absorbance were around 536 nm. Then the intensity
increased till 10 min without any change in the peak wavelength,
which was corresponding to the results published earlier [30].
The presence of polysaccharides and flavoniods in C. auriculata
leaves must have influenced the formation of gold nanoparticles
and the antidiabetic potential [31]. The stabilization of nanoparti-
cles and the functional groups of the compounds responsible were
identified using FT-IR studies and the results were similar to the
biosynthesis carried out earlier to obtain AuNPs [32]. Shiv Shankar
et al. reported that the formation of pure metallic nanoparticles and
bimetallic nanoparticles by reduction of the metal ions is possibly
facilitated by reducing sugars and/or terpenoids present in Neem
leaf broth [33]. The presence of extracellular polysaccharides and
tannins may facilitate the stabilization of nanoparticles [34] and
plant leaf has high level of oxalic acid and its oxalate can also act
as reducing agent [35]. The carbonyl groups from the amino acid
residues and peptides of proteins have a strong affinity to bind met-
als; hence the protein can act as encapsulating agent and protect
the nanoparticles from agglomeration [36].
The exact nature and size of the AuNPs formed can be stud-
ied by measuring the XRD. The XRD spectrum measured in this
case resulted in three intense peaks observed in the spectrum
which agrees to the Bragg’s reflection of AuNPs. The average size of
the AuNPs was calculated using Debye–Scherrer as reported ear-
lier [37]. The results obtained from the TEM study gives a clear
indication regarding the shape and size of the nanoparticles. The
AuNPs formed were predominantly monodisperse with diameter of
15–25 nm. On careful observation of various magnifications of TEM
images of gold nanoparticles it is noted that the particles are of uni-
form size ca. around 17 nm. The results obtained from the EDAX give
a clear indication regarding the other elements involved in the syn-
thesis of nanoparticles. The presence of some carbon and oxygen
along with gold signals suggests the presence of biomolecules in the
gold nanoparticles, which were not observed in many biosynthesis
of nanoparticles. This infers the presence of stabilizing molecules
on nanoparticles. On studying the effect of pH, no major change in
the wavelength confirmed the stability of the nanoparticles syn-
thesized in the present investigation [38]. This property of stability
maintenance at a wide range of pH infers the possibility to use C.
auriculata mediated AuNPs in drug delivery applications and syn-
thesis is termed as facile since synthesis of gold nanoparticles is
carried out using a too easy method.
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V. Ganesh Kumar et al. / Colloids and Surfaces B: Biointerfaces 87 (2011) 159–163
5. Conclusion
Facile synthesis of AuNPs using antidiabetic potent plant
resulted in gold nanoparticles of average size 15–25 nm, stable at
a wide range of pH [3.4–10.2] which may have the potential to be
used in antidiabetic treatment. In future, selection of such plants
with therapeutic value will create a new platform to utilize the
potential of herbal medicine in nanoscience for drug delivery or
biomedical applications.
Acknowledgements
We thank DST-Nanomission, Government of India for its
financial support for the project (SR/NM/NS-06/2009) and the man-
agement of Sathyabama University, Chennai for its stanch support
in research activities.
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