Journal of Alloys and Compounds 430 (2007) 330–333

Synthesis of nanocrystalline hydroxyapatite by using precipitation method

I. Mobasherpour a,∗ , M. Soulati Heshajin b , A. Kazemzadeh a , M. Zakeri c
a Ceramics Department, Materials and Energy Research Center, P.O. Box 31787-316, Tehran, Iran
b Biomaterials Engineering Department, Amir Kabir University of Tehran, Tehran, Iran
c Materials Engineering Department, Islamic Azad University of Saveh, Saveh, Iran

Received 12 February 2006; received in revised form 1 May 2006; accepted 4 May 2006
Available online 23 June 2006

Abstract
In this investigation, hydroxyapatite powder has been synthesized from the calcium nitrate hydrated and di-ammonium hydrogen phosphate
solution by precipitation method and heat treatment of hydroxyapatite powders. In order to study the structural evolution, the Fourier transform
infrared spectroscopy (FTIR), the X-ray diffraction (XRD) and simultaneous thermal analysis (STA) were used. Transmission electron microscopy
(TEM) and scanning electron microscopy (SEM) were used to estimate the particle size of the powder and observe the morphology and agglomeration
state of the powder. Results show that hydroxyapatite nanocrystalline can successfully be produced by precipitation technique from raw materials.
Hydroxyapatite grain gradually increased in size when temperature increased from 100 to 1200 ◦ C, and the hydroxyapatite hexagonal-dipyramidal
phase was not transformed to the other calcium phosphates phases up to 1200 ◦ C.
© 2006 Elsevier B.V. All rights reserved.

Keywords: Hydroxyapatite; Precipitation; Nanocrystallization; X-ray diffraction

1. Introduction upon the technique, materials with various morphology, stoi-

Hydroxyapatite (HAP: Ca10 (PO4 )6 (OH)2 ) has attracted
much attention as a substitute material for damaged teeth or
bones over the past several decades, because of its crystallo-
graphical and chemical similarity with various calcified tissues
of vertebrates [1,2].
Hydroxyapatite is the main mineral constituent of teeth and
bones. HAP ceramics does not exhibit any cytoxic effects. It
shows excellent biocompatibility with hard tissues and also with
skin and muscle tissues. Moreover, HAP can directly bond to
the bone. Unfortunately, due to low reliability, especially in wet
environments, the HAP cannot presently be used for heavy load-
bearing applications, like artificial teeth or bones [1].
Multiple techniques have been used for preparation of HAP
powders, as reviewed in several works [3]. Two main ways for
preparation of HAP powders are wet methods and solid state
reactions. In the case of HAP fabrication, the wet method can be
divided into three groups: precipitation, hydrothermal technique,
and hydrolysis of other calcium phosphates [3,4]. Depending
∗ Corresponding author. Tel.: +98 261 6204131.
E-mail address: I Mabasherpour@merc.ac.ir (I. Mobasherpour).
chiometry, and level of crystallinity can be obtained. Solid state
reactions usually give a stoichiometric and well-crystallized
product, but they require relatively high temperatures and long
heat treatment times. Moreover, sinterability of such powders is
usually low. In the case of precipitation, where the temperature
does not exceed 100 ◦ C, nanometric-size crystals can be pre-
pared. They have shapes of blades, needles, rods, or equiaxed
particles. Their crystallinity and Ca/P ratio depend strongly upon
the preparation conditions and are in many cases lower than
those for well-crystallized stoichiometric hydroxyapatite. The
hydrothermal technique usually gives HAP materials with a high
degree of crystallinity and with a Ca/P close to the stoichiometric
value. Their crystal size is in the range of nanometers to millime-
ters. Hydrolysis of tricalcium phosphate, monetite, brushite, or
octacalcium phosphate requires low temperatures and results in
HAP needles or blades having the size of microns. However, in
most cases, the hydrolysis product is highly nonstoichiometric
[1].
It is possible to improve the properties of HAP ceramic by
controlling important parameters of powder precursors such as
particle size and shape, particle distribution and agglomeration
[5]. Nanocrystalline HAP powders exhibit greater surface area
[6]. It can provide improved sinterability and enhanced densifi-

0925-8388/$ – see front matter © 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.jallcom.2006.05.018
 I. Mobasherpour et al. / Journal of Alloys and Compounds 430 (2007) 330–333 331

cation to reduce sintering temperature [7]. Moreover, nanometer

sized HAP is also expected to have better bioactivity than coarser
crystals [8,9], nanophase ceramics clearly represent a unique
and promising class of orthopedic/dental implant formulations
with improved osseointegrative properties [9,10]. Recently, the
use of precipitation processes for synthesis of HAP becomes an
important research objective [11].
In this investigation, the precipitation method has been
adapted to synthesize nanocrystalline HAP powder. Powder
characterization including phase composition, morphology and
distribution of grain size has been performed. In this method
offers a molecular-level mixing of the calcium and phosphorus
precursors, which is capable of improving chemical homogene-
ity and increasing phase transformation temperature to the other Fig. 1. FTIR spectroscopy analysis of the powder after drying at 80 ◦ C.
calcium phosphates phase of resulting HAP in comparison with
conventional method.
The DTA and TGA (STA) curves for the hydroxyapatite pow-
2. Experimental der after drying are illustrated in Fig. 2. The first endothermic
region range from 90 to 295 ◦ C with a peak at about 250 ◦ C,
Hydroxyapatite compounds were prepared by solution-precipitation method which corresponds to the dehydration of the precipitating com-
using Ca(NO3 )2 ·4H2 O (Analar No. 10305) and (NH4 )2 HPO4 (Merck No. 1205) plex and the loss of physically adsorbed water molecules of the
as starting materials and ammonia solution as agents for pH adjustment. A sus-
hydroxyapatite powder. The weight loss in this region is 16%.
pension of 0.24 M Ca(NO3 )2 ·4H2 O (23.61 g Ca(NO3 )2 ·4H2 O in 350 ml distilled
water) was vigorously stirred and its temperature was maintained at 25 ◦ C. A With increasing temperature from 295 to 1200 ◦ C no peak has
solution of 0.29 M (NH4 )2 HPO4 (7.92 g (NH4 )2 HPO4 in 250 ml distilled water) been observed, except a weight loss of 6% is observed at the
was slowly added dropwise to the Ca(NO3 )2 ·4H2 O solution. In all experiments TGA curve in the temperature range which is assumed to be
the pH of Ca(NO3 )2 ·4H2 O solution by ammonia solution was 11. This can be resulted gradual dehydroxyllin in hydroxyapatite powder. This
explained by the following reaction:
can be explained by the following reaction [12]:
10Ca(NO3 )2 ·4H2 O + 6(NH4 )2 HPO4 + 8NH4 OH Ca10 (PO4 )6 (OH)2 → Ca10 (PO4 )6 (OH)2−2x Ox x + xH2 O
→ Ca10 (PO4 )6 (OH)2 + 20NH4 NO3 + 20H2 O
In order to investigate the effect of heat treatment on nanocrys-
In the next step the precipitated HAP has been removed from solution by cen- tallization and phase transformation in hydroxyapatite powder
trifugation method at the rotation speed of 3000 rpm. The resulting powder was by precipitation method. Fig. 3 presents the XRD spectra at
dried at 80 ◦ C and then calcined at 100, 450, 900 and 1200 ◦ C for 1 h. various temperatures for hydroxyapatite calcined powders by
The structure of the resulting powder after drying was evaluated by Fourier precipitation method. Particularly noteworthy, the hydroxyap-
transform infrared (FT-IR) spectroscopy. The phase transformation during heat
atite peaks gradually increased in intensity when the sample
treatment and crystallite size evolutions were carried out by X-ray diffraction
analysis using a siemens diffractometer (30 kV and 25 mA) with Cu K␣ radiation was heated from 100, 450, 900, up to 1200 ◦ C (1 h holding time
(λ = 1.5405 Å). The scherrer method was used to evaluate the crystallite size. at each temperature), indicating further nucleation/growth of the
DTA and TG were performed simultaneously in an STA (Polymer Labora- hexagonal-dipyramidal nanocrystals inside the powder particles.
tories PL-1640) using alumina pans as sample holders. The sample used was in At room temperature the stable phase is hexagonal-dipyramidal.
bulk and weighed 8 g. The heating rate was 10 ◦ C/min and air was used as the
This hexagonal-phase-growth XRD data with increasing tem-
purging gas.
The morphology of the selected resulting powder was examined with a Cam- perature is reported here for the first time, but the hexagonal-
bridge scanning electron microscope (SEM) operating at 25 kV.
Finally, transmission electron microscopy (TEM) was used in characterizing
the particles. For this purpose, particles were deposited onto Cu grids, which
support a “holey” carbon film. The particles were deposited onto the support
grids by deposition from a dilute suspension in acetone or ethanol. The particle
shapes and sizes were characterized by diffraction (amplitude) contrast and, for
crystalline materials, by high resolution (phase contrast) imaging.

3. Results and discussion

The structure of the powder was analyzed using FT-IR spec-

troscopy after dried at 80 ◦ C, as shown in Fig. 1. In the FT-IR
analysis, mainly the peaks for PO4 3− and OH− groups in the
hydroxyapatite can be identified in Fig. 1. Peaks at 560–610 and
1000–1100 cm−1 must be due to PO4 3− . For the OH− group the
peak positions are 636 and 3572 cm−1 . Fig. 2. DTA and TGA traces of the hydroxyapatite powder after drying at 80 ◦ C.
332 I. Mobasherpour et al. / Journal of Alloys and Compounds 430 (2007) 330–333
Fig. 3. XRD patterns of heat treatment hydroxyapatite powders for different
temperatures.
dipyramidal phase was not transformed to the other phases up
to 1200 ◦ C.
The patterns due to the as-prepared HAP at 100 ◦ C bears with
it the characteristic patterns of HAP but not with much resolution
and intensity. It contains no other crystalline phase other than
HAP. The broad patterns around at (2 1 1) and (0 0 2) indicate
that the crystallites are very tiny in nature with much atomic
oscillations. The XRD patterns of the heat treatment HAP at
450 and 900 ◦ C show increase in intensity due to planes around
(2 1 1), (0 0 2), (3 0 1), (2 2 2) and (2 1 3). Again it rules out the
formation of any new crystalline phase other than HAP. Again
at the heat treatment temperature of 1200 ◦ C, new crystalline
phases are not seen. This precludes even amorphous phases, as
the patterns appear very well resolved. As the increase in inten-
sity is noticed for every pattern with increase in heat treatment
temperature.
The decomposition of HAP partly into tricalcium phosphate
was also reported by Raynaud et al. [13]. This decomposi-
tion into tricalcium phosphate was also not observed in the
present study when the HAP was heat treatment at 1200 ◦ C.
Reported transformation of HAP into oxyapatite between 1200
and 1400 ◦ C [14], but at 1200 ◦ C no such transformation is
observed in the present study.
The apparent size of hydroxyapatite crystallites obtained
from XRD profile analysis by Scherrer method [15–17] is shown
in Table 1 and Fig. 4. In this method of broadening contribution
due to the crystallites size are taken into account. A gradual
increase in crystallite size was observed for hydroxyapatite with
increasing in heat treatment temperature.
The microstructure of the powder prepared by the present
process after calcined at 1200 ◦ C for 1 h, examined by SEM,
Table 1
Calculated crystallite size from XRD pattern by Scherrer method
Heat treatment temperature (◦ C) Grain size (nm)
100 7.75
450 8.01
900 27.79
1200 59.06
Fig. 4. The hydroxyapatite crystallites obtained from XRD profile analysis by
Scherrer method.
as shown in Fig. 5. As it can be seen from the morphologies
of particles, there is a distribution of small particles and large
agglomerates. These agglomerates consist of very fine particles
that are cold welded together.
TEM micrographs of the hydroxyapatite powders before heat
treatment and after heat treatment are seen in Figs. 6 and 7.
The microstructure of the hydroxyapatite particles after dry-
ing is observed to be almost like spherical, with particle size
in the range 8–20 nm. When temperature increases to 1200 ◦ C,
the particle size of hydroxyapatite is found to be 40–50 nm.
Although the morphology of the particles is observed to be
almost like irregular hexagonal. When the sample was heated
from 100 to 1200 ◦ C, indicating further nucleation/growth of
the nanocrystals inside the powder. The exact particle nucleation
and growth mechanisms are not clear. Nucleation occurs proba-
bly by either hydrolytic reactions or a salting-out phenomenon.
Growth could be via diffusive molecular deposition or an aggre-
gation of primary particles by increase temperature. This TEM
observation on the shape of particles agrees with our earlier XRD
studies.
R2HAP
0.96
0.92
0.99
0.98 Fig. 5. SEM micrograph of hydroxyapatite particles after calcined at 1200 ◦ C
for 1 h.
 I. Mobasherpour et al. / Journal of Alloys and Compounds 430 (2007) 330–333
Fig. 6. TEM micrograph of the hydroxyapatite powder before heat treatment.
Fig. 7. TEM micrograph of the hydroxyapatite powder after heat treatment.
4. Conclusions
Hydroxyapatite compound with nanoparticle can success-
fully be produced by precipitation technique from Ca(NO3 )2 ·
4H2 O and (NH4 )2 HPO4 solution as starting materials for
333
the first time. Heat treatment was carried out on hydrox-
yapatite samples with different temperature. The hydroxyap-
atite grain gradually increased in size when the sample was
heated from 100 to 1200 ◦ C, and the hydroxyapatite hexagonal-
dipyramidal phase was not transformed to the other calcium
phosphates phases up to 1200 ◦ C. In contrast, after heat treat-
ment hydroxyapatite particles obtained from the homogeneous
precipitation are perfect monodispersed with sizes ranging from
nanometers.
Acknowledgments
The authors would like to acknowledge Mr. S. Razavi for
helping in preparing of this paper and Mr. Kaviani for doing
XRD experiments.
References
[1] W. Suchanek, M. Yoshimura, J. Mater. Res. 13 (1) (1998).
[2] L.L. Hench, J. Am. Ceram. Soc. 74 (1991) 1487.
[3] H. Aoki, Science and Medical Applications of Hydroxyapatite, JAAS,
Tokyo, 1991.
[4] R.Z. LeGeros, Calcium Phosphate in Oral Biology and Medicine, Karger
AG, 1991.
[5] S. Best, W. Bonfield, J. Mater. Sci. Mater. Med. 5 (1994) 516.
[6] R. Legeros, Clin. Mater. 14 (1993) 65.
[7] K.C.B. Yeong, J. Wang, S.C. Ng, Mater. Lett. 38 (1999) 208.
[8] S.I. Stupp, G.W. Ciegler, J. Biomed. Mater. Res. 26 (1992) 169.
[9] T.J. Webster, C. Ergun, R.H. Doremus, R.W. Siegel, R. Bizios, Biomaterials
22 (2001) 1327.
[10] T.J. Webster, R.W. Siegel, R. Bizios, Biomaterials 20 (1999) 1221.
[11] Y. Han, S. Li, X. Wang, X. Chen, Mater. Res. Bull. 39 (2004) 25–
32.
[12] M.G.S. Murray, J. Wang, C.B. Pontoon, P.M. Marquis, J. Mater. Sci. 30
(1995) 3061.
[13] S. Raynaud, E. Champion, D. Bernache-Assollant, P. Thomas, Biomaterials
23 (2002) 1065.
[14] J. Zhou, J. Chen, X. Zhang, K. De Groot, J. Mater. Sci. Mater. Med. 4
(1993) 83.
[15] B.D. Cullity, Elements of X-ray Diffraction, second ed., Addison-Wesley
Publishing, 1977.
[16] G.K. Williamson, W.H. Hall, Acta Metall. (January) (1953) 22–31.
[17] M. Zakeri, R. Yazdani-Rad, M.H. Enayati, M.R. Rahimipour, J. Alloys
Compd. 403 (2005) 258–291.