Available online at www.sciencedirect.

com

Procedia Engineering 42 (2012) 1816 – 1821

20th International Congress of Chemical and Process Engineering CHISA 2012

25 – 29 August 2012, Prague, Czech Republic

Synthesis of sphere-like nanoparticles of hydroxyapatite

T. Dědourkováa a*, J. Zelenkab, M. Zelenkováb,L. Benešc, L. Svobodaa
a
University of Pardubice, FCHT, Department of Inorganic Technology, Doubravice 41, 532 10 Pardubice, Czech Republic
b
Synpo, a.s., S. K. Neumanna 1316, 532 07 Pardubice, Czech Republic
c
University of Pardubice, FCHT, Joint Laboratory of Solid State Chemistry, Studentská 84, 532 10 Pardubice, Czech Republic

Abstract

Hydroxyapatite due to its excellent biocompatibility, bioactivity and osteoconductivity is widely used in orthopedic,
dental, and maxillofacial applications for bone reconstruction. Nanoparticles of hydroxyapatite, which are prepared
synthetically, have chemical and structural similarity with nature hydroxyapatite of bones and teeth. Sphere-like
nanoparticles of hydroxyapatite were synthesized by wet chemical precipitation method under atmospheric pressure,
using calcium nitrate and diammonium phosphate as reactants and ammonia was chosen as pH adjusting agent. The
crystal morphology and particle size were analyzed by atomic force microscopy and X-ray disc centrifuge system.
Composition and chemical structure were identified by X-ray diffraction, Fourier transform infrared spectroscopy and
thermogravimetric analysis.

© 2012 Published by Elsevier Ltd. Selection under responsibility of the Congress Scientific Committee
(Petr Kluson) Open access under CC BY-NC-ND license.

Keywords: Nanoparticles; hydroxyapatite; biomedical application

1. Introduction

The natural bone is a composite mainly consisted of nano-sized hydroxyapatite crystals well arranged
within the polymeric matrix of collagen type I. The mineral part of human bone is especially made of
nano-sized carbonated hydroxyapatite crystals, which represent 65 % of its total weight [1].
Synthetic nano-hydroxyapatite (n-HAp) is known to be one of the most important implantable
materials because of its biocompatibility, bioactivity and osteoconductivity coming from the analogy to

* Corresponding author. Tel.: +420466037171; fax: +420466037275.

E-mail address: st10009@student.upce.cz.

1877-7058 © 2012 Published by Elsevier Ltd. Open access under CC BY-NC-ND license.
doi:10.1016/j.proeng.2012.07.576
 T. Dědourková et al. / Procedia Engineering 42 (2012) 1816 – 1821 1817

the mineral components of natural bones that is why it is used as a substitute material for human hard
tissues. Hydroxyapatite has attracted much attention as a material for orthopedic, dental and maxillofacial
applications for bone reconstruction [2, 3]. Synthetic n-HAp exhibits strong affinity to host hard tissues
due to the chemical similarity between n-HAp and mineralized bone of human tissue. The recent trend in
bioceramic research is focused on improving mechanical and biological properties of hydroxyapatite
using nanotechnology because of poor mechanical properties [4].

2. Experiment

Nanoparticles of hydroxyapatite were prepared by wet chemical precipitation method. 0.3 M water
solution of calcium nitrate (Ca(NO3)2.4H2O, Penta, Czech Republic) and 0.3 M water solution of
diammonium phosphate ((NH4)2HPO4, Lachema, Czech Republic) were used as initial solutions. Solution
of (NH4)2HPO4 was added drop by drop in solution of Ca(NO 3)2 under vigorous stirring at 40 °C.
Ammonia was used for adjustment of pH value. Suspension was stirred 1.5 h after addition of initial
solution. The resultant suspension was filtered and washed with distilled water to remove the residual
impurities. The final product was atmospherically dried at room temperature.
The crystal morphology and particle size were analyzed by atomic force microscopy (AFM, Scanning
Peak Microscopy, Dimension Icon, Bruker, Germany) under ambient conditions by mode Peak Force
QNM (Quantitative NanoMechanics). The silicon nitrid probe having a resonant frequency 45-95 kHz, a
force constant 0.4 N/m and a radius of curvature 2 nm was used.
Particle size was studied by X-ray disc centrifuge system (BI-XDC Particle Sizer, Brookhaven
Instruments Corporation, USA), speed of disc rotation was 600 rpm. The XRD pattern was recorded
using X-ray diffraction (D8 ADVANCE Diffractometer, Bruker AXS, Germany) with CuK α radiation (λ
= 1.5418 Å). The sample was further characterized by Fourier transform infrared spectroscopy (FTIR
Spectroscope Nicolet IMPACT 400D, USA) in the range of 400-4000 cm-1 using KBr pellet technique.
Thermogravimetric analysis (TGA Q500, TA Instruments, USA) of the material was performed between
25-900 °C in air at a heating rate of 10 °C per minutes.

3. Results and discussion

The AFM images of n-HAp are shown in Fig.1 and Fig. 2. The size of hydroxyapatite crystal was
nano-grade. The sphere-like primary crystals of n-HAp were 30-50 nm in diameter and some aggregates
could be observed.
XDC method was used for measurement of particle size. This method measures sedimentation of
nanoparticles in a gravitational or a centrifugal field. X-rays from a low power X-ray tube are passed
through the disc. The intensity of the transmitted beam is measured with a detector consisting of a
scintillation counter whose output is recorded by the computer as a function of time. Curve of particle
size distribution can be observed in Fig.3. Determined n-HAp particle size distribution was 240-410 nm.
It is evident difference between results from AFM and XDC methods. XDC showed bigger size of
nanoparticles because of possible formation of aggregates.
XRD diffractogram (Fig. 4) confirmed hexagonal structure of hydroxyapatite with P63/m space group
and cell dimensions of a = b = 9.43 Å; c = 6.88. Å.
FTIR analysis, which is shown in Fig. 5, displayed specific vibrations for bands present in
hydroxyapatite crystals. Phosphate groups were detected at 960 cm-1, 1033 cm-1, 575 cm-1 and 601 cm-1;
OH- band was identified at 3575 cm-1 inside the H2O region. Furthermore, carbonate bands were
observed at 1425 cm-1 and 875 cm-1. X-ray diffraction and infrared spectroscopy confirmed structure of
hydroxyapatite. These carbonate substitutions of hydroxyl or phosphate groups appeared during wet
1818 T. Dědourková et al. / Procedia Engineering 42 (2012) 1816 – 1821

chemical precipitation because of ambient carbon dioxide solubilization related to vigorous stirring. This
modification of stoichiometric n-HAp is similar to natural n-HAp in bones.
TGA analysis showed 8.5 % loss of weight, which was a result of removal of adsorbed and possible
lattice water, decarboxylation of HAp or condensation of HPO42- releasing water, as can be seen from Fig.
6.

Fig. 1. AFM image of n-HAp on mica, primary nanoparticles 30-50 nm, map of adhesion, scan 2 μm

Fig. 2. AFM image of n-HAp on mica, aggregates (A: 140x120 nm, B: 214x100 nm, C: 190x170 nm), topography, scan 2 μm
 T. Dědourková et al. / Procedia Engineering 42 (2012) 1816 – 1821 1819

110

100

90

differential
80

70

60

50

40

30

20

10

0,2 0,3 0,4 0,5

Pm]
d [P

Fig. 3. Particle size distribution of sphere-like nanoparticles

HA14
350
d=2,817

340
330
320
310
300
290
280
270
260
250
85
2,7

240
d=3,450

d=

230
220
d=2,723

210
Lin (Counts)

200
190
180
170
160
150
d=1,843

140
d=1,946

130
120
d=2,635

110
100
d=2,266

d=1,724

90
d=1,782
d=1,808
d=3,087

d=1,757

80
d=8,174

d=1,893
d=3,178

70
d=1,456
d=1,505

60
d=2,301

d=2,065
d=2,155
d=2,533

d=1,434
d=3,892

d=1,475
d=5,277

d=4,084

d=1,646

d=1,544

50
d=2,004

d=1,613

40
30
20
10
0

5 10 20 30 40 50 60 7

2-Theta - Scale
HA14 - File: 12189-KAnT.RAW - Type: 2Th/Th locked - Start: 5.000 ° - End: 70.000 ° - Step: 0.020 ° - Step time: 4. s - Temp.: 25 °C (Room) - Time Started: 17 s - 2-Theta: 5.000 ° - Theta: 2.500 ° - Phi: 0.00 ° - Aux1: 0.0 -
Operations: Fourier 20.000 x 1 | Strip kAlpha2 0.500 | Import
04-007-5086 (A) - Hydroxylapatite, syn - Ca5(PO4)3(OH) - d x by: 1. - Hexagonal - P63/m (176) - a 9.43200 - b 9.43200 - c 6.88100 - alpha 90.000 - beta 90.000 - gamma 120.000 - Primitive - 2 - 530.139 - WL: 1.5406 -

Fig. 4. X-ray diffraction of powder of n-HAp

1820 T. Dědourková et al. / Procedia Engineering 42 (2012) 1816 – 1821

Fig. 5. FTIR spectrum of n-HAp

100

99

98
weight [%]

97

96

95

94

93

92

91

0 100 200 300 400 500 600 700 800 900

T [°C]

Fig. 6. TGA analysis of nano-powder of HAp

4. Conclusions

This work was focused on preparing sphere-like nanoparticles of hydroxyapatite. Nanoparticles were
successfully prepared via wet chemical precipitation method using calcium nitrate tetrahydrate and
diammonium phosphate as precursors. Shape, size and chemical structure of nano-hydroxyapatite ware
 T. Dědourková et al. / Procedia Engineering 42 (2012) 1816 – 1821 1821

studied. AFM showed that the primary particles have spherical shape and diameter 30-50 nm. FTIR
spectra and XRD diffractogram confirmed structure of hydroxyapatite.

Acknowledgements

This work was supported by IGA University of Pardubice in terms of project SG FChT04.

References

[1] Cengiz B, Gokce Y, Yildiz N, Aktaz Z, Calimli A. Synthesis and characterization of hydroxyapatite nanoparticles. Colloids
Surf A; Physicochem Eng Asp 2008;322:29-33.
[2] Catros S, Guillemot F, Lebraud E, Chanseau C, Perez S, Bareille R, Amédée J, Fricain JC. Physico-chemical and biological
properties of a nano-hydroxyapatite powder synthesized at room temperature. IRBM 2010;31:226-33.
[3] Sanosh KP, Chu MC, Balakrishnan A, Lee YJ, Kim TN, Cho SJ.Synthesis of nano hydroxyapatite powder that simulate teeth
particle morphology and composition. Curr Appl Phys 2009;9:1459-62.
[4] Kuriakose TA, Kalkura SN, Palanchamy M, Arivuoli D, Dierks K, Bocelli G, Betzel C. Synthesis of stoichiometric nano
crystalline hydroxyapatite by ethanol-based sol-gel technique at low temperature. J Cryst Growth 2004;263:517-23.