Spectrochimica Acta Part A 74 (2009) 1127–1134

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Spectrochimica Acta Part A: Molecular and

Biomolecular Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Spectroscopic investigations on the synthesis of nano-hydroxyapatite from

calcined eggshell by hydrothermal method using cationic surfactant as template
K. Prabakaran ∗ , S. Rajeswari
Department of Analytical Chemistry, University of Madras, Guindy Campus, Chennai, Tamilnadu 600 025, India

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

Article history: The present work reports the successful synthesis of nano-hydroxyapatite, Ca10 (PO4 )6 (OH)2 (denoted
Received 5 January 2009 HAP) from calcined eggshell by hydrothermal method using cationic surfactant (CTAB) as regulator of
Received in revised form 13 August 2009 nucleation and crystal growth. The reaction involved in the synthesis was studied elaborately. The influ-
Accepted 12 September 2009
ence of reaction temperature, ageing time and CTAB concentration on the synthesis of nano-HAP are also
studied in addition to the effect of sintering temperature on the crystal growth. Spectral characterization
Keywords:
involving Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) techniques were
Nano-hydroxyapatite
performed for functional group analysis and phase identification of the materials, respectively. Thermal
Eggshell
Cationic surfactant
stability of nano-HAP was investigated by thermal analysis (TG/DTA). The physical characteristics, such
Hydrothermal method as morphology and particle size of the synthesized nano-HAP were assessed thoroughly by scanning
electron microscopy (SEM) and atomic force microscopy (AFM) techniques. The results have revealed
that well-crystallized nano-HAP was synthesized by hydrothermal treatment at 160 ◦ C for 10 h with the
addition of CTAB at critical micelle concentration (CMC). It was also found that the synthesized nano-HAP
was thermally stable up to 1100 ◦ C.
© 2009 Elsevier B.V. All rights reserved.

1. Introduction Although the chemical synthesis of HAP came back to several

Hydroxyapatite [HAP, chemical formula Ca10 (PO4 )6 (OH)2 )] has
attracted attention of researchers during the past three decades as
an implant material because of its excellent biocompatibility and
bioactivity [1,2]. It is the most important member of phosphate
mineral group; mainly because it is the prototype of the inorganic
material in hard tissues such as bone and teeth and is also regarded
as an important implant material with significant clinical poten-
tial. Thus much research concerned with HAP has been carried out
in fields such as crystallography, medical science and material sci-
ence. Clinically, HAP has been used in a variety of physical forms;
as a sintered ceramic, such as in granular, block or porous form, as a
deposited coating, such as bioactive coating on a bioinert implant,
or as filler [3,4]. The properties of HAP powder, such as crystallinity,
morphology, surface area and particle size of the resulting HAP
powder will determine the effectiveness of the powder in its spe-
cific applications [5]. For applications where HAP powders sintered
to form a ceramic, the mechanical properties of HAP will be depen-
dant on the sintered density and microstructure of the final sintered
product [6–13]. Hence it is essential to understand the way in which
HAP powder synthesized to form ceramic.
∗ Corresponding author. Tel.: +91 44 2220 2719; fax: +91 44 2235 2494.
E-mail address: chitrapraba07@gmail.com (K. Prabakaran).
decades ago, the synthetic methodology has been developing over
the years in order to satisfy the requirements for various biomedi-
cal applications. Synthetic HAP is normally prepared either by wet
chemical route [14,15] or by solid state reaction method [16]. These
conventional methods, however, mostly prepare irregular forms
of powders with microsized particles. However, the hydrothermal
method is very versatile for the synthesis of nanophase mate-
rials and has been well established. Bone itself is a composite
consisting of HAP nano-rods embedded in the collagen matrix.
Hence nano-sized HAP particles are desirable when biocompatibil-
ity is considered. Moreover, new developments on the production
of nano-sized HAP particles have led to many new applications.
For example, nano-sized HAP particles can retard multiplication
of cancer cells and be used as an efficient drug delivery agent.
Hydrothermal method, which has proved to be a convenient way
to prepare nanophase materials, including salts, metal oxides, etc.,
has also been applied for the synthesis of nano-sized HAP parti-
cles, but the control on morphology (which largely determines the
behaviour of a certain material) is very poor.
It has been proved that the organized organic surface can control
the nucleation of inorganic materials by geometric, electrostatic,
and stereochemical complementarity between incipient nuclei and
functionalized substrates [17]. In the presence of extraneous addi-
tives, however, the role of crystal surface is more analogous to
the conventional view of host-guest systems. In such case, the
surface layers of the crystal can incorporate soluble additives pro-

1386-1425/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.saa.2009.09.021
1128 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134
vided there is a degree of complementarity in charge and size
between the guest ions and interstices in the structure of the
crystal boundary layers. However, the samples prepared at ambi-
ent temperature were generally poorly crystallized so that their
strength was not high. Therefore, it is believed that the combina-
tion of the two methods might give a better result. In the present
study, we explore the assistance of surfactant in HAP preparation
by hydrothermal method from eggshells, a waste material. Calcined
eggshell and phosphoric acid were used as calcium and phospho-
rous sources, respectively. A cationic surfactant, cetyltrimethyl or
hexadecyltrimethylammonium bromide (CTAB), which is widely
used to achieve control of morphology in many aqueous synthetic
methods, was chosen as the regulator to control the crystalliza-
tion of HAP. The influence of reaction temperature, ageing time,
CTAB concentration and sintering temperature on the nucleation
and crystal growth process is investigated.
2. Materials and methods
2.1. Powder preparation
The surface area of the eggshells collected were mechanically
cleaned using double distilled water and were dried in an air oven
for 1 h at 100 ◦ C. The dried eggshells were ground into fine powder
and calcined at 900 ◦ C for 1 h. The amount of calcium present in
calcined eggshell was estimated by conventional EDTA titrimetric
method. A stoichiometric amount of the calcined eggshell powder
was dispersed in doubly distilled water (pH∼12). To the phospho-
ric acid solution (0.075 M), different concentrations of CTAB was
added and kept overnight (about 8 h) to ensure the co-operative
interaction and self-assembly process was completed. The phos-
phate solution was then added drop wise to calcium suspension at
a controlled flow rate under continuous but constant stirring. After
total addition of the reactants the suspension was transferred to a
stainless steel autoclave, sealed tightly and hydrothermally treated
at 160 ◦ C for 10 h. The resultant precipitate was thoroughly washed
with double distilled water, filtered and dried at 80 ◦ C for 2 h in an
air oven. The dried precipitate was sintered at 400, 700 and 900 ◦ C,
respectively for 2 h in an air furnace.
2.2. Powder characterization
FT-IR spectral analysis was performed for functional group anal-
ysis of the samples by using Shimadzu model 8300. The FT-IR
transmittance spectra of the samples were obtained in the range
400–4000 cm−1 using KBr pellet technique. All the samples were
identified by their X-ray diffraction pattern (XRD) using RICH-
SEIFERT model 3000 X-ray diffractometer with Cu K␣ (␭ = 1.5406 Å)
incident radiation. The diffraction patterns were taken to the 2
ranges from 20◦ to 60◦ at a step sampling interval of 0.02◦ . Iden-
tification of phases were achieved by comparing the diffraction
pattern of HAP with JCPDS standards of card no: 04-0932 for
HAP and 04-0733 for Ca(OH)2 . The size of the elementary crys-
tallites was calculated using Scherrer’s formula. The full-width at
half-maximum of the 0 0 2 reflection was taken to measure the
crystallite sizes of the HAP powder.
The scanning electron microscopy (SEM) and energy disper-
sive X-ray microanalysis (EDAX) analysis were performed for the
observation of microstructure and chemical constituent changes of
the materials. The instrument Philips 501 scanning electron micro-
scope equipped with EDAX was used for the microstructural and
microanalysis. The samples were coated with a thin layer of gold
using an Edwards sputter coater S150B instrument. The EDAX spec-
tra are acquired at an accelerating voltage of 100 kV. The elemental
composition (to provide Ca/P molar ratio of a selected area of the
sample) was determined by EDAX. Atomic force microscopy (AFM)
to study the surface topography of the samples was performed
using a Picoscan 2100 (Molecular Imaging, USA) employing Au
coated silicon nitride pyramidal probes with tip size of ∼30 nm.
The as-prepared samples were ground into a fine powder and were
uniaxially pressed using a stainless steel die at 125 MPa to form
a pellet of size 10 mm × 0.3 mm diameter and the pressed bodies
were sintered at 900 ◦ C for 2 h for analyzing the surface topogra-
phy using AFM. The weight loss and thermal stability of the samples
were also evaluated from the thermogravimetric analysis data. An
HETZSCH-Geratebau GmBH thermal analyser having STA (simulta-
neous thermal analyzer) 409C microbalance with a heating rate of
10 ◦ C min−1 was employed.
3. Results and discussion
3.1. Synthesis of HAP
The present study employs hydrothermal method for the pro-
duction of HAP using calcined eggshell as calcium source and
phosphoric acid as phosphorous source. The thermal treatment
used for eliminating the organic component of eggshell at 900 ◦ C
(1 h) converts calcite (CaCO3 ) into calcium oxide by freeing carbon
dioxide (CO2 ). The expected reaction is
CaCO3 → CaO + CO2 ↑ (1)
Fig. 1 shows the FT-IR spectra of raw, 800 and 900 ◦ C cal-
cined eggshells, respectively. The FT-IR spectra shown in Fig. 1a
and b indicate characteristics peaks corresponding to CO3 2− ion at
1420–1570 and 875 cm−1 . The carbonate ion peak for 800 ◦ C cal-
cined eggshell was found to be intense when compared to the raw
eggshell and this elicits the evolution of free calcium carbonate after
calcination. The characteristic PO4 3− and OH− absorption bands
of HAP at 570, 600, 1033, 1085 and 3572 cm−1 , respectively were
also observed for all samples. Dennis et al. [18] highlighted the fact
that some cuticular vesicles of eggshell contain phosphate min-
eral HAP (deposited by lipoprotein and protein–lipid complexes of
eggshell) and may be involved in the termination of calcite crys-
tal growth. However, the intensities of these peaks are very less
since amount of HAP present in the crystal is very minimum (less
than 1%). The bending mode of water is positioned at 1650 and
3440 cm−1 . It is very intense for raw eggshell and less intense for
the eggshell calcined at 800 and 900 ◦ C. It is also evident from the
Fig. 1c that the ␯3 carbonate absorption band around 1400 cm−1
and ␯2 vibrational mode absorption at 875 cm−1 got disappeared
for the sample calcined at 900 ◦ C, which indicates the complete
elimination of carbonate ion at this temperature.
Fig. 2 shows the XRD pattern of eggshell calcined at 800 and
900 ◦ C. The CaCO3 was completely decomposed and turned to CaO
at about 900 ◦ C. The phase analysis was performed using JCPDS card
no. 05-0586 for CaCO3 and 37–1497 for CaO. It is very obvious from
the figure that no other secondary phases were present except CaO
(Fig. 2) and CaCO3 (Fig. 2). Thus, the formation of CaO inferred from
the corresponding XRD pattern elicits the loss of gaseous CO2 from
CaCO3 at 900 ◦ C.
The formed calcium oxide when dispersed in double distilled
water produces calcium hydroxide (confirmed by XRD shown in
Fig. 2). The reaction can be represented as follows.
CaO + H2 O → Ca(OH)2 (2)
The calcium hydroxide thus produced acts as one of the initial
reactant (calcium source), which on reacting with orthophospho-
ric acid to form HAP. The reaction theoretically expected for the
formation of HAP is
10Ca(OH)2 + 6H3 PO4 → Ca10 (PO4 )6 (OH)2 + 18H2 O (3)
 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134 1129

Fig. 2. XRD pattern of (a) and (b) eggshell sintered at 800 and 900 ◦ C, respectively;
(c) Ca(OH)2 .

The morphological changes of HAP precipitates subjected to

Fig. 1. FT-IR spectra of (a) raw eggshell, (b) eggshell calcined at 800 ◦ C and (c)
eggshell calcined at 900 ◦ C.
hydrothermal treatment at different ageing time (say, 1, 5, 10
and 15 h) were observed by SEM technique (see supplementary
information). It is then noted that samples subjected to hydrother-
mal treatment for 1 and 5 h formed large aggregates, but for those
samples treated 10 and 15 h exhibited uniformly nano-sized HAP

It is noted that there is no by-product derived in the above reac-

tion except water. Thus phase pure HAP can be produced by this
method.
The effect of reaction temperatures was examined and the XRD
patterns of HAP precipitates synthesized at different temperatures
are shown in Fig. 3. All the samples were found to comprise of
HAP (JCPDS card No. 09-432) phase. The diffraction spectrum at
2 = 32◦ consisted of two peaks, and the diffraction peaks at 2 = 33◦ ,
34◦ and 49.5◦ , respectively, became narrower and sharper, and the
diffraction peaks at 2 = 26◦ , 28.8◦ and 48.1◦ began to appear with
increasing reaction temperatures. These suggested that higher tem-
perature produced HAP with larger proportion of crystalline phase.
The FT-IR spectra of HAP samples synthesized at 80, 160 and 200 ◦ C
are shown in Fig. 4. The intensity of the OH stretching vibration
at 3571 cm−1 and OH structural vibration at 631 cm−1 for 200 ◦ C
heated samples was higher when compared to the sample heated at
160 ◦ C. This suggested that higher temperature produced HAP with
larger proportion of crystalline phase, which was consistent with
the results from the XRD patterns as shown in Fig. 3. The absorption
peaks at 2920 and 2851 cm−1 were assigned to CH3 and CH2 groups,
respectively, got eliminated when the sample heated at 200 ◦ C. This
implied that the amount of CTAB was lower in samples synthesized
at higher temperature. Hence, the heating temperature at 160 ◦ C Fig. 3. XRD patterns of HAP synthesized at the different temperature for 10 h with
was chosen as the optimum temperature for HAP synthesis. the CTAB (at CMC).
1130 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134
Fig. 4. FT-IR spectra of HAP synthesized using CTAB at the different temperatures for 10 h.
about same size. So it is reasonable to propose that when the reac-
tion time is long enough, the final products would be invariably
nano. Hence 10 h was chosen as the optimum ageing time for HAP
synthesis.
The effect of CTAB on HAP phases is shown in Fig. 5. The OH
absorption peaks at 3570 and 630 cm−1 , OH stretching vibration
and OH structural vibration, decreased and disappeared with the
decrease in CTAB into the phosphate solution (Fig. 5a and b). The
amount of CTAB remaining in the calcium phosphate samples
was higher for CTAB at critical micelle concentration (CMC) with
CTAB:PO4 3− ratio = 1:1 as indicated by the intensity of absorption
peaks at 2920 and 2851 cm−1 , and the intensity of OH absorp-
tion bands got increased, which implied that CTAB concentrations
below CMC could reduce the crystallization of HAP. The SEM pic-
tures of HAP synthesized at CMC and below CMC of CTAB (see
supplementary information) confirmed that the synthesis at CMC
improves crystallinity of the HAP powder. The results are in good
agreement with the results observed in FT-IR. Thus, it can be con-
cluded that the micelles act as nucleating points for the growth of
HAP crystals and hence CMC with CTAB:PO4 3− ratio = 1:1 was cho-
sen as the optimum CTAB concentration for the synthesis of highly
crystalline HAP.
3.2. Effect of sintering temperature
Fig. 6 shows FT-IR absorption spectra of HAP powders heated
at 80, 400, 700 and 900 ◦ C. The infrared band positions and their
assignments are summarized in Table 1. All the spectral sam-
ples show vibrational modes corresponding to the phosphate and
hydroxyl except carbonate groups. A broad envelope between
3200 and 3800 cm−1 due to the O–H stretch of water and HAP
was observed for the sample heated at 80 ◦ C. The O–H groups
are hydrogen bonded. The peak at around 1650 cm−1 assigned to
bending mode of water was clearly visible for the sample heated
at 80 ◦ C. However, the intensities of these peaks were found to
decrease for those samples heated at higher temperatures. The
sharp peak at 3572 cm−1 observed for those samples heated at
higher temperatures is assigned to unhydrogen bonded free O–H
 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134 1131

Fig. 5. FT-IR spectra of HAP synthesized at 160 ◦ C for 10 h with the different
CTAB:PO4 3− ratio (a) 0.01:1, (b) 0.1:1 and (c) 1:1 (CTAB at CMC).

stretch which may be present at the surface of the crystallites. The

peaks at 630 and 3572 cm−1 assigned to characteristic OH group
peaks for stoichiometric HAP was well distinguished for all the
samples heated at different temperatures. This implies that HAP
synthesized by this method exhibits greater crystallinity and high Fig. 6. FT-IR spectra of HAP synthesized by hydrothermal method heated at (a) 80,
crystalline HAP powder was formed even at 80 ◦ C. Carbonate bands (b) 400, (c) 700 and (d) 900 ◦ C.
are usually identified as sharps peaks at around 1420, 1480 and
875 cm−1 . According to van der Houwen et al. [19], these peaks
cess was carried out in alkaline range (pH 10.5). However, it is
are attributed to the dissolving of atmospheric CO2 as the pro-
interesting to note that the CO3 2− ion substitution is completely
absent in all the four samples including the sample heated at low
Table 1 temperature. The reason could be attributed to the closed reaction
Assignments of the observed vibrational frequencies of HAP heated at various condition of hydrothermal method during synthesis, which pre-
temperatures. vents the interference of atmospheric CO2 with the reactants. All
Assignments Observed vibrational frequencies (cm−1 ) the four vibrational modes of phosphate ion (␯1 , ␯2 , ␯3 and ␯4 )
are IR active and are observed in HAP spectra for samples heated
Heating temperatures
at different temperatures. The ␯1 and ␯3 phosphate bands in the
80 ◦ C 400 ◦ C 700 ◦ C 900 ◦ C region of 900–1100 cm−1 and ␯4 absorption bands in the region of
PO4 bend ␯4 565 563 562 562 500–700 cm−1 are used to characterize apatite structure. The spec-
PO4 bend ␯4 603 605 602 602 tral bands in the range 900–1100 cm−1 containing symmetric ␯1
Structural OH 632 631 631 630 and asymmetric ␯3 , P–O stretching modes of the phosphate groups
CO3 2− group – – – –
were observed. The symmetric P–O stretching mode for HAP occurs
PO4 stretch ␯1 962 962 961 962
PO4 bend ␯3 1028 1035 1042 1050 at 962 cm−1 , which indicates typical HAP structure. The stretching
PO4 bend ␯3 1090 1093 1098 1099 and the bending modes of PO4 3− appeared at 602 and 562 cm−1 as
CO3 2− group (␯3 ) – – – – intense sharp peaks for all the samples, which elicits the crystalline
CO3 2− group (␯3 ) – – – – nature of the samples. van der Houwen et al. [19] reported identifi-
H2 O adsorbed (␯2 ) 1628 1636 1624 1630
H2 O adsorbed 3418 3448 3426 3441
cation of HPO4 2− based on its water absorption bands at 1210 cm−1
Structural OH 3572 3570 3572 3572 together with one at 1130 cm−1 (shoulder to the absorption band
OH stretch 3740 3744 3744 3744 at 1099 cm−1 ) and a clear absorption band at 879 cm−1 . But these
CH3 group 2920 2921 – – features are not clearly evident in the spectrum, as the absorption
CH2 group 2851 2851 – –
band due to PO4 3− of HAP is intense and broad. It is also interest-
1132 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134

Fig. 7. Thermal analysis of HAP synthesized by hydrothermal method.

ing to note that the absorption peaks corresponding to CH3 and

CH2 groups of CTAB at 2920 and 2851 cm−1 was present for the
sample heated at 80 ◦ C as intense peaks. However, the intensity
of these peaks was decreased for the sample heated at 400 ◦ C. But
these peaks were not present for the samples sintered at higher
temperatures. The reason could be attributed to the elimination of
CTAB from HAP for the samples sintered beyond 400 ◦ C. The result
was well accompanied by the TG-DTA results, where an exothermic
peak was observed in DTA curve at around 500 ◦ C.
The thermal stability of HAP samples was investigated by
TGA/DTA. There are four regions in the TGA curves of the syn-
thesized HAP powders as shown in Fig. 7. These four regions can
be assigned based on the corresponding trend derived from DTA
results. Region I (30–200 ◦ C) corresponded to the loss of water
present in the HAP samples as reflected by the weight loss in
TGA curve and exothermic peak in DTA curve at 110 ◦ C. Region Fig. 8. XRD pattern of HAP synthesized by hydrothermal method heated at (a) 80,
II (200–600 ◦ C) signified the decomposition of CTAB present in (b) 400, (c) 700, (d) 900 and (e) 1100 ◦ C.
the HAP samples, exhibiting a large weight loss and prominent
The degree of crystallinity influences the bioresorbability and the
exothermal peak in DTA profile between 500 and 600 ◦ C. This
biological behavior of HAP. Recent studies have shown that more
suggested that CTAB could be readily removed from HAP sam-
crystalline structure induced less resorbability of HAP. With the
ples when sintered at higher temperatures. There were minimal
increase in sintering temperature these diffraction peaks become
changes in region III (600–700 ◦ C) and a small weight loss in region
sharp, which indicates the increase in crystallinity of HAP powder
IV (700–1050 ◦ C). Beyond this region, an exothermic peak in DTA
synthesized by this method. It can be inferred from these results
was observed at around 1100 ◦ C due to the decomposition of cal-
that the increase in temperature causes increase in crystallinity of
cium deficient HAP (CDHA) into HAP and ␤-TCP according to Eq. (4).
HAP. The sintering of HAP improves the crystallinity and density as
Thus, the TG-DTA results have revealed that the HAP was stable up
well.
to around 1100 ◦ C.
Heating HAP samples from room temperature to higher temper-
Ca(10−x) (HPO4 )x (PO4 )(6−x) (OH)(2−x) → Ca10 (PO4 )6 (OH)2 ature causes changes in phase transformations, loss of physically
and chemically bound H2 O molecules and OH groups occurs. This
+ Ca3 (PO4 )2 , where (0 ≤ x ≤ 1) (4)
was proved from the TGA curves in the interval of 100–800 ◦ C. The
Fig. 8 shows the XRD patterns of HAP powder synthesized by above quoted changes during the synthesis and processing of the
hydrothermal method heated at various temperatures. It can be HAP precipitate precursor display good reproducibility. The present
noted from figures that all the samples resulted in diffraction peaks investigation shows that HAP powder heated at 1150 ◦ C exhibits
corresponded only to the standard for HAP except the sample sin- other ␤-TCP in addition to HAP, which was attributed to the decom-
tered at 1150 ◦ C. Broad diffraction peaks were observed for the HAP position of HAP at this particular temperature. The results are in
powders heated at low temperatures (80 ◦ C). After sintering the good agreement with the TG-DTA results where an exothermic
material for 2 h at 400 ◦ C, better crystalline HAP is observed and peak was observed at this temperature.
the same trend is preserved for the sample heated at 900 ◦ C for The degree of crystallinity influences the bioresorbability and
2 h. The sharp reflection peaks appeared in the range 31.7–52.5◦ the biological behaviour of HAP. Recent studies have shown that
represents the characteristic peak of apatite phase. Reflections at more crystalline structure induced less resorbability of HAP. With
(0 0 2), (1 1 2), (3 0 0) and (2 1 1) planes are observed for powders the increase in sintering temperature these diffraction peaks
sintered at 900 ◦ C, which suggests that depending upon the sinter- become sharp, indicating increase in crystallinity of HAP powder.
ing temperature, the transformation of amorphous to crystalline The fraction of crystalline phase (Xc ) in the HAP powders was eval-
HAP phases are possible. The patterns demonstrated the stable uated by the following equation [20]:
nature of HAP with no extraneous peaks, indicating the formation  
V11 2/3 0 0
of stoichiometric HAP with Ca/P molar ratio of 1.67. It suggests that Xc = 1 − (5)
I3 0 0
pure monophasic crystalline HAP phase can be produced at 900 ◦ C.
 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134
Table 2
Elemental composition of nano-HAP powder.
Element Weight % Atom %
O 41.76
Na 0.18
Mg 0.23
P 18.15
Ca 39.18
C 0.50
Total 100.00 100.00
where I3 0 0 is the intensity of (3 0 0) diffraction peak and V1 1 2/3 0 0
is the intensity of the hollow between (1 1 2) and (3 0 0) diffraction
peaks of HAP. The evaluated degrees of crystallinity for the samples
heated at 80 and 900 ◦ C was found to be 0.94 and 0.99. It is inferred
from the resulted values that the increase in temperature improves
crystallinity of HAP.
The peak broadening of XRD reflection can be used to estimate
the crystallite size in a direction perpendicular to the crystallo-
graphic plane based on Sherrer’s formula as follows [21,22]:
0.9
Xs =
(ˇ cos )
where Xs is the crystallite size (nm);  the wavelength of monochro-
matic X-ray beam (nm) ( = 0.15406 nm for Cu K␣ radiation); ˇ the
full-width half-maximum for the diffraction peak under consid-
eration (rad); and  the diffraction angle (◦ ). The reflection plane
(0 0 2) was chosen for calculation of the crystallite size since the
peak appears at this Miller’s family plane is sharper and isolated
from others and shows the crystal growth along the c-axis of the
HAP crystalline structure. The calculated crystallite size, Xs for HAP
powder heated at 80 ◦ C and 900 ◦ C at this diffraction peak using
Scherrer’s formula was found to be 34 and 88 nm, respectively.
Thus, both the XRD and TG-DTA results have depicted that the sta-
bility of the HAP synthesized by hydrothermal method using CTAB
as a regulator was extended up to 1100 ◦ C and thereafter it was
decomposed into ␤-TCP.
SEM pictures of HAP powder heated at different tempera-
tures revealed that the synthesized powder appeared to be white
crystals, bouncy and free flowing powder (see supplementary
information). The initial powders consist of small agglomerated
rod-like crystals, which upon sintering became deagglomerated
and appeared as nano-rods. The morphology of HAP powder
sintered at 900 ◦ C indicates that most of the HAP particles are non-
agglomerated and monodispersive. It can be inferred that majority
of the particles are of single crystals with regular shape and cleaner
contours. The phase constitution and chemical homogeneity of HAP
were examined by EDAX analysis. Table 2 illustrates the elemen-
tal composition of the synthesized nano-HAP powder. The Ca/P
molar ratio was found to be 1.67, which indicates the formation
of stoichiometric HAP. The result shows that the HAP is primarily
composed of Ca, P and O along with the trace amount of Na, Mg and
C.
The AFM images (both 2D and 3D view) of the HAP synthe-
sized using CTAB by hydrothermal method have allowed making
a morphological interpretation of the crystal growth occurred after
sintering at 900 ◦ C (see supplementary information). The topogra-
phy of the HAP crystals is relatively smooth with a height variation
of ∼2.5 nm. The image shows the relevant variations in the crystal
growth recorded on 0.2 ␮m × 0.2 ␮m area of the sintered HAP pel-
let. Both the 3D view and the 2D view depict the presence of several
plateaus with approximately same heights. Especially the 3D view
illustrates the maximum height of the plateau, which in turn elic-
its the height of the crystal growth. The crystals were observed to
present well-defined grain boundaries due to non-agglomeration
1133
of the particles during sintering. The particle sizes are ranging from
100 to 300 nm and are largely agglomerated. The result is in good
agreement with the results obtained from SEM analysis. The reason
61.31 for the formation of nano-HAP with controlled microstructure can
0.18 be explained as follows.
0.12 The addition of surfactant causes the controlled nucleation and
14.29 crystallization of the HAP during synthesis. The behavior of CTAB
23.85
was considered to correlate with the charge and stereochemistry
0.25
properties. In an aqueous system, CTAB would ionize completely
and result in a cation with tetrahedral structure and forms micelles
at CMC. These micelles act as nucleating points for the growth of
HAP crystals with controlled structure. Meanwhile, the phosphate
anion is also possessing tetrahedral structure. It has been reported
by several authors that a process called molecule recognition could
be realized at the inorganic/organic interface by the charge and
stereochemistry complementarity. Hence it can be proposed that
the same charge and structure complementarity. The added surfac-
tant was then supposed to bind to certain faces of crystals and bind
to certain ions as well, so that ions could be incorporated to the
existing nuclei at a steady rate and the final shape and size could
be well controlled. Further, the anisotropy of CTAB itself probably
induced the axis orientation growth of HAP.
(6) The high temperature and pressure of hydrothermal treatment
could exert both effects on the final product. Firstly, they might
cause some fibers to aggregate, but the interaction between the
aggregates is so weak that they tend to dissolve in the aqueous
system. Secondly, they might raise the solubility of HAP to some
extent and accelerate the dissolution and crystallization process.
Since the crystallization process is under critical control of CTAB,
the resulting HAP was invariably nano-sized.
4. Conclusions
In the present work, nano-HAP with the particle size ranging
from about 100–300 nm was synthesized by hydrothermal method
from calcined eggshell using CTAB as regulator. Functional group
and phase identification corresponding to eggshell and HAP were
investigated by FT-IR and XRD studies. SEM and AFM techniques
(see supplementary information) employed to study the mor-
phology of HAP have exhibited the formation of well-crystallized
nano-particles with uniform shape. TG/DTA results have shown
that the formed nano-HAP was thermally stable up to 1100 ◦ C.
The Ca/P ratio obtained from EDAX analysis confirmed the value of
1.67 corresponding to stoichiometric HAP. The observations further
revealed that the crystallization process under critical micelle con-
centration (CMC) of CTAB resulted in the production of nano-HAP
with uniform size. Moreover, the use of eggshell as a calcium source
represents a potential diminution in the costs of HAP production
with the additional advantage of recycling a waste product.
Acknowledgement
The authors acknowledge the financial support rendered by
University Grants Commission (UGC) through University With
Potential For Excellence (UWPFE) programme to carryout this
work.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.saa.2009.09.021.
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