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Utilization of eggshell waste as raw material for synthesis of hydroxyapatite

Article  in  Colloid and Polymer Science · June 2015

DOI: 10.1007/s00396-015-3646-0

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Colloid Polym Sci
DOI 10.1007/s00396-015-3646-0
ORIGINAL CONTRIBUTION
Utilization of eggshell waste as raw material for synthesis
of hydroxyapatite
Sarute Ummartyotin 1 & Benchamaporn Tangnorawich 1
Received: 2 May 2015 / Revised: 24 May 2015 / Accepted: 27 May 2015
# Springer-Verlag Berlin Heidelberg 2015
Abstract Calcium oxide was successfully derived from egg-
shell waste. It was employed as a calcium precursor in order to
prepare hydroxyapatite. It can be noted that eggshell presented
high thermal stability, and it was calcined from 700 to 1000 °C
for 3 h for calcium oxide formation. Morphological properties
were investigated, and it presented the irregular shape with a
variation on size. The feasibility of calcium oxide for hy-
droxyapatite preparation was evaluated by the chemical pre-
cipitation with phosphoric acid. X-ray diffraction confirmed
that single phase of hydroxyapatite was obtained.
Keywords Eggshell waste . Calcium oxide . Hydroxyapatite
Introduction
In recent years, the utilization of bio-based materials with the
value-added concept has been extensively developed. Numer-
ous efforts have been improved on the use of bio-based mate-
rials with any versatility on application [1–3]. Up to the pres-
ent time, many aspects of bio-based materials have been
researched. One of the most effective bio-based materials
was focused on the development of bio-based polymer. For
example, cellulose and its derivatives have been extensively
Sarute Ummartyotin and Benchamaporn Tangnorawich contributed
equally to this work.
* Sarute Ummartyotin
sarute.ummartyotin@gmail.com
* Benchamaporn Tangnorawich
bencha@tu.ac.th
1
Department of Physics, Faculty of Science and Technology,
Thammasat University, Patumtani, Thailand
developed for many purposes of application [4–6]. It can be
employed in many sectors of application such as automotive
part, infrastructure, as well as electronic device [7–9]. The
other approach was considered on chitosan and its derivatives.
It can be employed in active packaging, drug delivery system,
as well as wound dressing [10, 11]. The attempt on the use of
bio-based materials was extent to polylactic acid,
polybutylene succinate, as well as starch. They were devel-
oped and investigated on the feasibility to be utilized in many
industrial sectors such as bio-cup, film for active packaging,
and tray for food. On the other hand, the development on bio-
based ceramic has been extensively pushed for both industrial
and academic researches. From the fundamental point of view,
bio-based ceramic was defined as ceramic derived from natu-
rally occurring resource. It can be involved not only on bio-
based ceramic but also on bio-based glass. The main applica-
tion on bio-based ceramic was related to biocompatibility for
human such as bone repairing, skin permeation, as well as bio-
based implant for organ and any tissue engineering. It was
important to note that bio-based ceramic was related to inert
material for human body and to the other extreme of resorba-
ble material.
In order to fulfill this criterion, one of the most effective
bio-based ceramic was considered on hydroxyapatite (HA)
[12, 13]. It was important to note that hydroxyapatite was
structurally formulated as Ca10(PO4)6(OH)2, and it gained
many attracted attentions on the feasibility as an implant ma-
terial because of its excellent biocompatibility and bioactivity
[14, 15]. Due to the involvement of the member of phosphate
mineral group, it can be therefore employed as the prototype
of the inorganic material for hard tissue engineering such as
bone and teeth, and subsequently, it was considered as an
effective resource for implant material with significant clinical
technology. Therefore, up to the present time, hydroxyapatite
gained many interests exponentially from materials science to

an approach on medical technology including drug delivery
system, organ implant and repair, as well as the further ap-
proaches on dental materials [16, 17].
Moreover, in order to encourage on the use of waste-based
product with higher efficiency, the concept of value-added
should be strongly performed. Eggshell was considered as
an effective source for calcium. It was commonly known that
egg was commonly considered as food which was versatile in
many products such as cake, desserts, as well as soup. How-
ever, due to the growth on population, the production and
consumption on food, especially on egg, were considered as
an important issue. Also, due to the requirement on production
of egg, eggshell was subsequently similarly increased and it
was considered as disposal or waste. It was therefore more
difficult to manage [18, 19].
In order to support on the value-added concept with sus-
tainable and renewable aspect, eggshell was therefore being
pushed to be researched as the effective source of calcium. In
this research work, we wish to present the role of eggshell as
waste material as an effective source for hydroxyapatite syn-
thesis. Characterization on structure and properties was there-
fore investigated.
Experimental
Materials
Eggshell was collected from canteen, Faculty of Science and
Technology, Thammasat University. Sodium hydroxy, phos-
phoric acid, ammonia, and methanol of analytical grade were
purchased from Sigma-Aldrich, Thailand, and they were used
as received without further purification.
Methods
Synthesis and characterization of calcium oxide derived
from eggshell waste
Eggshell waste was collected from the canteen and it was
washed with methanol. After that, it was dried for 24 h in an
oven at 100 °C in order to eliminate contaminants and odor.
After that, it was milled and pressed through a sieve with a
25-μm mesh. Eggshell particle was investigated on its thermal
stability and morphological properties by thermogravimetric
analysis and scanning electron microscope, respectively. Fine
particle of eggshell was therefore calcined at 700, 800, 900,
and 1000 °C for 3 h. After that, it was evaluated on structure
and properties by X-ray diffraction, scanning electron micro-
scope, transmission electron microscope, UV–vis spectrosco-
py, Fourier transform infrared, and dynamic light scattering.
Finally, calcium oxide particle was obtained.
Colloid Polym Sci
Synthesis of hydroxyapatite from calcium oxide as a precursor
derived from eggshell
Hydroxyapatite was then synthesized using the calcium oxide
derived from eggshell and phosphoric acid by a wet chemical
precipitation technique. The molar ratio of calcium oxide and
phosphoric acid was estimated to be 2:1. During reaction,
calcium oxide was suspended into water and phosphoric acid
was added to the solution at a rate of 10 drops/min. The pH of
the solution was controlled to be greater than 10 by using
ammonium solution. The reaction was operated at 120 °C
for 6 h.
After the reaction was complete, the suspension was fil-
tered and washed with distilled water before being dried in
an oven at 80 °C for 24 h. The dried powder was milled and
pressed through a sieve with a 25-μm mesh to produce a
uniform particle. After that, it was investigated on structure
by X-ray diffraction.
Characterizations
X-ray diffraction
Calcium oxide particle was investigated on crystal structure
by X-ray diffraction (XRD, Phillips P.W. 1830 diffractome-
ter). It was employed by using nickel-filtered CuKa radiation.
Diffraction patterns were recorded over a range of 25–80.
Prior to investigation, the sample should be stored in a desic-
cator for moisture absorption prevention.
Scanning electron microscope
Eggshell and calcium oxide particles were investigated on
morphological properties by SEM (a JOEL JSM-6301F scan-
ning microscope). The machine was operated at an accelera-
tion voltage of 1.5 kV at a working distance of 15 mm to
identify the morphological properties of powders. Before in-
vestigation, the samples were sputter-coated with Au to en-
hance the electrical conductivity.
Transmission electron microscope
Eggshell and calcium oxide particles were investigated on
morphological properties by TEM, Hitachi H-7000. The
solution was suspended in methanol and dropped on a
molybdenum grid. After that, the grid was dried at
50 °C for methanol evaporation and kept into the TEM
chamber. The image thus obtained was processed with
computer for identification of the domains in which cer-
tain lattice fringes appear. For this purpose, TEM image
was captured under ×50,000 magnification. The accelera-
tion voltage of electron beam was set at 100 kV.
Colloid Polym Sci
Dynamic light scattering
Dynamic light scattering (DLS) experiments were performed
with a light scattering apparatus from Malvern Instruments
(Model 4700), with an argon laser source (wavelength
520 nm). DLS measurements were made at a scattering angle
of 90 with a 200-mm pinhole to obtain the homodyne inten-
sity–intensity correlation.
Thermogravimetric analysis
Thermal degradation behavior of the eggshell particle was
determined by thermogravimetric analysis (TA-5000 TGA,
TA Instruments, DE, USA). Each sample (10 mg) was heated
with a heating rate of 10 °C/min in nitrogen atmosphere from
30 to 800 °C. The TGA balance flow meter was set at 20 psi
N2, while the purge flow meter was adjusted to 20 psi of
synthetic air.
Fourier transforms infrared spectroscopy
Fourier transform infrared (FTIR) was performed on a Bruker
Vector 22 mid-IR spectroscopy (Bruker, Germany); FTIR ab-
sorption spectra was recorded over 4000–400 cm−1 wavenum-
ber region at a resolution of 8 cm−1 with 1024 scans using a
deuterated triglycine sulfate (DTGS) detector. A straight line
between two lowest points in the respective spectra region was
chosen as a baseline. Potassium bromide (KBr) acting as a
non-absorbing medium was mixed with a solid sample (0.3–
0.5 wt%) by an agate mortar and pestle to prepare a pellet
specimen.
UV–vis spectroscopy
A Varian Cary 5000 UV–vis NIR spectrophotometer (Agilent
Technologies, CA, USA), equipped with transmittance acces-
sory, was employed to record the electronic spectrum of the
samples over the wavelength of 200–900 nm. This technique
allowed the study of the absorbance spectra of the samples.
The accessory consisted of a 110-nm diameter integrating
sphere and an in-built high-performance photomultiplier.
Each sample was placed in a sample cell specifically designed
for the instrument. A baseline was recorded and calibrated
using a polytetrafluoroethylene (PTFE) reference cell.
Results and discussion
Characterization on calcium oxide derived from eggshell
Prior to preparation of hydroxyapatite, eggshell was consid-
ered as an effective source of calcium. It was washed with
methanol and dried for 24 h at 100 °C in order to eliminate
contaminants and odor. From the structural point of view, it
was noted that the main composition of eggshell was calcium
carbonate with many sizes of particle. The structure was con-
sidered as a network of calcium carbonate with hierarchical
feature. In order to gain calcium oxide from calcium carbon-
ate, it was important to note that the decomposition of calcium
carbonate was related to calcium oxide formation. With the
increment on temperature, the decomposition of calcium car-
bonate can effectively provide calcium oxide. The chemical
equation of decomposition of calcium carbonate was related
to exothermic reaction, which was referred to CaCO3→CaO
and CO2 [18, 19].
Moreover, in order to evaluate the calcination temperature,
calcium carbonate derived from eggshell was investigated by
thermogravimetric analysis. Figure 1 exhibits the TGA ther-
mogram of eggshell. It can be found that the decomposition of
weight loss can be categorized into three different regions.
From room temperature to 600 °C, no weight loss was ob-
served, suggesting that no change on decomposition due to
increment of temperature. It was important to note that egg-
shell was facile to absorb water and any concern of humidity
might be detrimental on properties. To store eggshell in a
desiccator was one of the effective methods for water absorp-
tion prevention. Furthermore, at the temperature ranging from
600 to 800 °C, the wide region of weight loss was observed;
this was related to the heat combustion on eggshell or exother-
mic reaction. Significance on weight loss was due to the trans-
formation of calcium carbonate to calcium oxide. The loss of
weight was due to the evaporation of carbon dioxide gas.
After that, with the temperature from 800 °C to higher, no
weight loss was therefore observed, suggesting that complete
transformation was performed. To obtain calcium oxide from
eggshell, the calcination temperature should be set from
800 °C, and it was important to note that percent weight loss
was estimated to be 40 %, suggesting that calcium oxide can
be derived from eggshell only 40 % by weight.
Fig. 1 TGA thermogram of eggshell-based material
 Colloid Polym Sci

Figure 2 exhibits typical image on morphological proper-

ties of eggshell. It presented a cross-linked in 3D fiber-mesh
structure, similar to hierarchical feature. The diameter of fiber
was ranged from 10 nm–1 μm with a versatile on orientation.
It was remarkable to note that with the difference on size of
fiber and its hierarchical orientation, eggshell was therefore
considered as a high porosity material and it gain many inter-
ests on the development for membrane-based materials and
superabsorbent materials.
Figure 3 exhibits UV–vis spectra of the typical calcium
oxide derived from eggshell. The difference on calcination
on eggshell was therefore investigated. Weak absorption in
the visible light region was observed for all of the samples,
which was due to the uniformity on energy level or lattice
defects of calcium oxide derived from eggshell. It was impor- Fig. 3 UV–vis spectra of calcium oxide derived from eggshell: a 700 °C,
b 800 °C, c 900 °C, and d 1000 °C of calcination temperature
tant to note that the broad peak was observed in the UV region.
The peak was estimated to be 250 nm for all of the samples.
attributed to hydroxyl group. The vibration between O–H was
However, the intensity was different. This was probably due
therefore detected due to humidity absorption. The FTIR spec-
to the effect of crystallinity of calcium oxide. With the incre-
troscopy of calcined eggshells was also observed. Three fre-
ment on calcination of temperature, longer time was provided
quency bands inherent to the structure of CaO were clearly
and it was enough to be re-oriented. This experiment exhibited
illustrated. The broad bands at 1450 and 1000 cm−1 were
the similar trend as FTIR pattern. Absorption peak was there-
ascribed to the C–O stretching mode with respect to CO2
fore observed at the low region of wavelength.
adsorbed on the surface of CaO. However, it was important
The FTIR spectrum of calcined eggshells was investigated
to note that there is no different on result of FTIR spectra. The
as the preliminary experiment on chemical determination of
existence of peak was still similar for all experiments. Prelim-
calcium oxide derived from eggshell. Figure 4 exhibits FTIR
inary experiment on FTIR spectra can be indicated that calci-
pattern of calcium oxide derived from eggshell. It was impor-
um oxide was successfully derived from eggshell. More in-
tant to note that significant characteristic peaks at 1450, 900,
vestigation on chemical structure of eggshell should be pro-
and 700 cm−1 were corresponded to the carbonate group in
vided in order to evaluate the feasibility of eggshell as the
CaCO3, which was commonly referred to the main component
effective source for hydroxyapatite preparation.
of eggshell. Moreover, during measurement, due to high po-
The XRD experiment of calcium oxide was investigated.
rosity of calcium oxide, it was facile to absorb gas. Small
Figure 5 exhibits the XRD pattern of calcium oxide derived
frequency bands at 2900 and 2500 cm−1 were observed, and
from eggshell waste. In this experiment, variation on calcina-
they were attributed to the combination modes of different
tion temperature was investigated, and it was important to note
CO2. This discussion was strongly associated with previous
that predominant peak of CaCO3 was observed in the eggshell
article [20, 21]. Moreover, the peak position at 3650 cm−1 was

Fig. 4 FTIR pattern of calcium oxide derived from eggshell: a 700 °C, b
Fig. 2 Morphological properties of eggshell 800 °C, c 900 °C, and d 1000 °C of calcination temperature
Colloid Polym Sci
Fig. 5 XRD pattern of calcium oxide derived from eggshell: a 700 °C, b
800 °C, c 900 °C, and d 1000 °C of calcination temperature
(calcination temperature of 700 °C), suggesting that incom-
plete calcination was conducted. The existence of peak posi-
tion at 2θ of 28 was therefore observed, and it was investigated
as the calcium carbonate residual. From the fundamental point
of view, the main component of eggshell was referred to cal-
cium carbonate. With the increment on temperature, calcium
carbonate was therefore fired and it therefore obtained calcium
oxide. However, with the increment on calcination tempera-
ture to 800–1000 °C, no existence of calcium carbonate was
therefore observed, suggesting that complete calcination tem-
perature was operated. To prevent any residual, eggshell
should be investigated on its thermal stability. Moreover, in
Fig. 5, it can be noted that calcium oxide was formed in
Fig. 6 Morphological properties
of calcium oxide derived from
eggshell: a 700 °C, b 800 °C, c
900 °C, and d 1000 °C of
calcination temperature
between 800 and 1000 °C of calcination temperature. XRD
results indicated the single crystalline phase formation of
CaO. The strongest intensity CaO peak was observed at a 2θ
angle of 37°, which was corresponded to the (200) lattice
plane. At different calcination temperatures, the intensity of
the strongest CaO peak increased from 800 to 1000 °C. The
crystal size was estimated by the Scherrer formula: D=Kλ/
βcosθ, where D is crystallite size, K is a constant of 0.9, λ is
the X-ray wavelength, β is full width at half maximum
(FWHM), and θ is the diffraction peak. The (200) peak was
used to estimate the crystal, and it was found to be 70 nm.
Figure 6 exhibits the morphological properties of calcium
oxide derived from eggshell. The microstructure of calcium
oxide was presented with the magnification of ×10,000. It was
important to note that morphological properties of calcium
oxide were porous like with a variation on size. It exhibited
the irregular shape. With the calcination temperature of 700
and 800 °C, the presence of porous was due to the pathway of
solvent, water, and CO2 evaporation. It can be noted that re-
moval of CO2 can occur due to the increment on calcination
temperature. Moreover, with the calcination temperature of
900 and 1000 °C, removal of CO2 was therefore completed.
Growth of calcium oxide was subsequently performed. The
amount of porous was less; while the density was therefore
gained. It was important to note that appropriate calcination
temperature should be controlled due to the prevention on
agglomeration process.
After that, morphological properties of calcium oxide de-
rived from eggshell were therefore investigated by transmission
 Colloid Polym Sci

Fig. 7 Morphological properties

of calcium oxide derived from
eggshell: a 700 °C, b 800 °C, c
900 °C, and d 1000 °C of
calcination temperature

electron microscope. Figure 7 exhibits morphological proper- conducted at 800 °C. It should be noted that the role of calci-
ties of calcium oxide with a variation on calcination tempera- nation temperature should be optimized on the removal of re-
ture. The result was strongly associated with scanning electron sidual and the prevention of agglomeration among particles.
microscope with a high order of magnitude. Variation on size In the case of calcium oxide derived from eggshell, it was
and shape was therefore observed with the magnification of suspended in a water solution. It was important to note that
×50,000. The resolution was extend from microstructure to most of the particles were precipitates within a minute leaving
nanostructure level. However, due to the nature of calcium a clear aqueous supernatant immediately after the preparation.
oxide, it was facile to agglomerate. With agglomeration, the Table 1 exhibits the average diameter of calcium oxide particle
particle size was estimated to be 100 nm. Furthermore, it can with a standard deviation. The average particle size of calcium
be noted that all of the samples presented the crystallinity with a
spherical shape. The level of average particle was very high
with the increment on calcination due to the effect of material
formation or crystal growth. However, it is remarkable to note
that with the calcination temperature of 900 and 1000 °C, a
particle becomes agglomerate. To gain the most effective on
particle and its uniformity, calcination temperature should be

Table 1 Particle size distribution of calcium oxide derived from

eggshell

Sample Average particle Standard

size (nm) derivation (nm)

1. CaO calcined at 700 °C 196 12.3

2. CaO calcined at 800 °C 209 9.2
3. CaO calcined at 900 °C 249 11.5
4. CaO calcined at 1000 °C 662 67.5 Fig. 8 XRD pattern of hydroxyapatite prepared calcium oxide: a 700 °C,
b 800 °C, c 900 °C, and d 1000 °C of calcination temperature
Colloid Polym Sci
oxide was enhanced with increment on calcination tempera-
ture. It can be explained that with higher calcination temper-
ature, calcium oxide particle became agglomerate; it was
therefore presented as a bulky particle. However, it was com-
monly known that due to the nature of calcium oxide particle,
it presented the hydrophilic feature. To determine the particle
size of calcium oxide, the good distribution should be con-
trolled. This indicated that calcium oxide was facile to aggre-
gate in an aqueous suspension, suggesting that ultrasonic pro-
cess should be performed. It was important to note that with
the determination of particle of calcium oxide, good distribu-
tion should be performed in order to prevent any error from
measurement.
Preliminary investigation on the synthesis
of hydroxyapatite from eggshell
After calcium oxide preparation, the preliminary experiment as
the precursor source of hydroxyapatite was subsequently inves-
tigated. Impressive result was exhibited that calcium oxide de-
rived from eggshell presented the excellent performance on
hydroxyapatite formation. Figure 8 exhibits the XRD pattern
of as-synthesized hydroxyapatite powder. All patterns were
indexed to hexagonal hydroxyapatite. However, it can be noted
that the significance on intensity of peak was therefore observed
due to the variation on calcination temperature. In Fig. 8, it was
observed that the highest intensity peak existed at a 2θ of 32.
This result was associated with XRD standard of hydroxyapa-
tite (JCPDS: 01-084-1998). The crystallite size of the as-
synthesized hydroxyapatite powder was found to be a nano-
crystalline with a value of 45 nm. It can be noted that from
preliminary experiment, hydroxyapatite was successfully syn-
thesized for wet co-precipitation. Eggshell waste can be effec-
tively employed as a precursor for calcium source.
Conclusion
Calcium oxide was successfully derived from eggshell waste.
It was employed as a calcium precursor in order to prepare
hydroxyapatite. It can be noted that eggshell presented high
thermal stability, and it was calcined at different temperature
ranges from 700 to 1000 °C for 3 h. Morphological properties
were investigated, and it presented the irregular shape with a
variation on size. Furthermore, calcium oxide was employed
as an effective calcium for hydroxyapatite preparation. It was
successfully synthesized from wet chemical precipitation
route. X-ray diffraction confirmed that single phase of hy-
droxyapatite was obtained.
Acknowledgments The authors gratefully acknowledge the financial
support provided by Thammasat University under the TU Research
Scholar.
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