Microwave conversion of eggshells into ower-like hydroxyapatite nanostructure for

biomedical applications
G. Suresh Kumar
a
, A. Thamizhavel
b
, E.K. Girija
a,

a
Department of Physics, Periyar University, Salem 636 011, India
b
Department of Condensed Matter Physics, Tata Institute of Fundamental Research, Colaba, Mumbai 400 005, India
a b s t r a c t a r t i c l e i n f o
Article history:
Received 2 February 2012
Accepted 22 February 2012
Available online 28 February 2012
Keywords:
Biomaterials
Bioceramics
Eggshell
Hydroxyapatite
In the present work, the eggshell biowaste has been effectively utilized to produce ower-like hydroxyapa-
tite nanostructure by using a simple and rapid microwave irradiation method with the help of ethylenedia-
minetetraacetic acid (EDTA) as a chelating agent. Conversion of eggshell into HA occurs via the formation of
CaEDTA complex which subsequently reacted with phosphate under microwave irradiation. The obtained
product is Mg containing b-type carbonated HA and it can be a potential material for biomedical applications.
2012 Elsevier B.V. All rights reserved.
1. Introduction
Hydroxyapatite (HA, Ca
10
(PO
4
)
6
(OH)
2
) is the major inorganic
component of bone and teeth. It has been extensively used in various
forms for bone repairs and as coatings for metallic prostheses owing
to its biocompatibility, bioactivity and osteoconductivity [1]. Over
the past years, biologically derived natural materials such as corals,
nacre, sh bone, bovine bone, eggshells etc., have been converted
into useful biomaterials like HA and F-substituted HA [212]. These
materials often exhibit a ne-scale microstructure with variable po-
rosity, unusual crystal habits and morphologies, remarkable mechan-
ical and physical properties. Among the several natural calcium
resources, eggshell is a composite material consisting of calcium
carbonate (94%), calcium phosphate (1%), organic matter (4%) and
magnesium carbonate (1%) [8]. Everyday million tons of eggshells
are produced as biowaste around the world. Important issues in the
industrial process are minimization of wastes and recycling them
into useful products. Hence, developing a simple method to convert
these eggshells into HA is essential from the view point of material
science and engineering.
There are several reports on the synthesis of HA from eggshells
using direct or indirect conversion methods [812]. Direct conversion
process is done by time consuming hydrothermal method whereas
the indirect process involves the preparation of calcium precursor
from eggshells as a rst step and then the HA preparation by the
conventional methods like wet precipitation, sol-gel, microwave
irradiation, etc., as a second step [912]. On the other hand, HA
nanostructures with different morphologies have been synthesized
using organic modiers such as ethylenediaminetetraacetic acid
(EDTA), citric acid and polystyrene sulfonate [1315]. The aim of
the present work is to propose a simple and rapid microwave irradi-
ation method for recycling the eggshell biowaste to obtain ower-like
HA nanostructure using EDTA as a complexing agent.
2. Experimental
All the chemicals used for the experiments were analytical grade
obtained from Merck, India. Eggshells were collected and immersed
in boiling water to remove the inner membrane and surface contam-
inants. After drying, they were ground into powder by hand in a
mortar and immersed in sodium hypochlorite to remove organic
components. Then, they were extensively washed with Millipore
water and dried in vacuum oven for 5 h at 110 C. 1 g of eggshell
powder was then mixed with 0.1 M of EDTA solution to form Ca
EDTA complex. Then, 0.06 M of Na
2
HPO
4
solution was slowly added
with obtained CaEDTA complex and stirred for 30 min. The prelimi-
nary pH of the reaction mixture was about 8. The pH of the reaction
mixture was adjusted to 13 by the addition of NaOH solution. Then,
the prepared reaction mixture was put in a microwave oven
(2.45 GHz, 600 W, LG, India) and irradiated with microwave for
10 min. The white precipitate obtained was washed and dried in hot
air oven at 110 C for 5 h.
The X-ray diffraction (XRD) pattern of eggshell powder and syn-
thesized sample were obtained using Schimadzu XRD-6000 diffrac-
tometer in the range between 20 260 using Cu K radiation
(1.5406 ). FT-IR spectrum of synthesized sample was recorded in
Materials Letters 76 (2012) 198200
Corresponding author. Tel.: +91 9444391733; fax: +91 427 2345124.
E-mail address: girijaeaswaradas@gmail.com (E.K. Girija).
0167-577X/$ see front matter 2012 Elsevier B.V. All rights reserved.
doi:10.1016/j.matlet.2012.02.106
Contents lists available at SciVerse ScienceDirect
Materials Letters
j our nal homepage: www. el sevi er . com/ l ocat e/ mat l et
the 4000400 cm
1
region with 4 cm
1
resolution using a Perkin
Elmer RX1 FT-IR spectrometer by KBr pellet technique. The morphology
of eggshells and synthesized sample was examined using Zeiss Ultra
plus scanning electron microscope and the elemental analysis was
done using the Oxford INCA energy dispersive X-ray uorescence
(EDX) microanalysis.
3. Results and discussion
Fig. 1(a) shows the XRD pattern of eggshells and is in good agree-
ment with ICCD data for calcite (JCPDS le No. 05-0586). The average
crystallite size of the eggshell powder was calculated as 78 nm using
DebyeScherrer formula. SEM image (Fig. 1(b)) shows the surface
morphology of the eggshells. EDX analysis indicates the presence of
Ca, P, C, O and Mg in eggshells and the obtained elemental composi-
tion is shown in Table.1.
Fig. 2(a) shows the XRD pattern of synthesized powder. The X-ray
diffraction peaks matched well with ICDD standard data for HA
(JCPDS File No: 09-0432) which indicates the formation of phase
pure HA. The lattice constants and unit cell volume were calculated
as a=b=9.4509 , c=6.9270 and V=535.80
3
, respectively.
The FT-IR spectrum of the obtained product is shown in Fig. 2(b).
The characteristic PO
4
3
(
4
) vibrations of HA appeared at 563 and
601 cm
1
along with other
1
,
2
and
3
phosphate peaks at 472,
961, 1032 and 1108 cm
1
respectively. The bands at 3573 and
632 cm
1
are the characteristic OH stretching and liberational
modes of HA respectively [16]. The broad band extending from
2500 to 3600 cm
1
is attributed to the
3
and
1
stretching modes
of the H
2
O molecules. The band at 1642 cm
1
is attributed to the
2
bending mode of the H
2
O molecules [16]. Besides, it showed addition-
al peaks at 875 cm
1
(
2
), 1415 cm
1
(
3
) and 1459 cm
1
(
3
). The
positions of these peaks indicate the formation of b-type carbonated
HA [16].
SEM observation of the product obtained after microwave irradia-
tion is shown in Fig. 2(c). It exhibited typical ower-like
nanostructured HA. The ower-like nanostructure is composed of
leaf-like akes having 100200 nm width and 0.51 m length
which are extending radially from center. The corresponding EDX
taken from individual leaf-like ake indicates the presence of Ca, P,
O, C and Mg in the obtained product and its elemental composition
is shown in Table.1.
When the eggshell powder was introduced into EDTA solution it
dissolved with the evolution of carbonate according to the following
reaction:
CaCO
3
EDTA
4
CaEDTA
2
CO
3
2
When the CaEDTA solution was titrated against phosphate the
formation of a clear solution was observed. This is due to the en-
hanced stability of the CaEDTA complex as the working pH was
13. EDTA can form stable complex with calcium ions especially
under base condition. In the complex, EDTA acts as a hexadentate
unit by packaging itself around Ca
2+
with four atoms of oxygen
and two atoms of nitrogen and forms several chelate rings [13].
Under this condition, a three dimensional cluster of critical size
is formed which acts as nucleus for HA crystals and develop into
crystallite under the microwave irradiation. However further
growth of HA crystallites will be controlled by the stability of
CaEDTA complex as well as hindrance effect of adsorbed OH

ions on the facets of the crystallites. At pH 13, each facet of the

crystallite has almost the same probability to adsorb OH

ions
which generates limited active sites for the further growth. Now,
Ca will be incorporated into the active sites in form of CaEDTA
complex because of its higher stability. However the absorbed
CaEDTA complexes undergo decomposition under microwave
irradiation and HA crystals grow resulting in the formation of
ower-like nanostructure due to the anisotropic growth habit of
HA [13].
The presence of Mg was identied in the obtained HA, which was
inherited fromthe eggshells. Mg is known to be an important trace el-
ement in bone and teeth. Moreover, Mg plays an important role in
bone metabolism and its depletion causes bone fragility and bone
loss [1719]. Consequently, the incorporation of Mg ions into the
HA structure is of great interest for the development of articial
bones. Besides, Wang et al. reported that the ower-like HA acts as ef-
fective carriers for drug delivery applications [15]. The present study
shows that biowaste eggshells can be effectively utilized to obtain
ower-like HA nanostructure by using simple and rapid microwave
irradiation method.
4. Conclusion
Eggshell biowaste has been successfully recycled to obtain ower-
like HA nanostructure by using a simple and rapid microwave irradi-
ation method with the help of EDTA as a complexing agent. Conver-
sion of eggshell into HA occurs via the formation of CaEDTA
complex which subsequently reacted with phosphate under micro-
wave irradiation. The obtained product was identied as Mg contain-
ing b-type carbonated HA and it seems to be a promising material for
biomedical applications.
Table 1
Elemental composition of raw eggshells and the nal product obtained from EDX.
Samples Elemental composition (wt.%)
Ca P C O Mg
Eggshells 35.11 3.66 15.28 44.59 1.37
HA 25.34 15.40 7.61 50.61 1.05
Fig. 1. (a) XRD pattern and (b) SEM image of eggshells.
199 G.S. Kumar et al. / Materials Letters 76 (2012) 198200
Acknowledgments
The authors (G.S. and E.K.G) express their sincere thanks to
Department of Science and Technology (DST), India (Project Ref.
No: SR/FTP/PS-24/2009) for nancial support.
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Fig. 2. (a) XRD pattern, (b) FT-IR spectrum and (c) SEM images of the obtained product.
200 G.S. Kumar et al. / Materials Letters 76 (2012) 198200