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Synthesis of nano-hydroxyapatite under a sonochemical/hydrothermal condition

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2008 Biomed. Mater. 3 025002

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IOP PUBLISHING BIOMEDICAL MATERIALS
Biomed. Mater. 3 (2008) 025002 (6pp) doi:10.1088/1748-6041/3/2/025002

Synthesis of nano-hydroxyapatite under a

sonochemical/hydrothermal condition
S A Manafi, B Yazdani, M R Rahimiopour, S K Sadrnezhaad, M H Amin
and M Razavi
Ceramic Department, Materials and Energy Research Center, PO Box 14155-4777, Tehran, Iran
E-mail: rahimi40@yahoo.com

Received 23 October 2007

Accepted for publication 12 February 2008
Published 25 March 2008
Online at stacks.iop.org/BMM/3/025002

Abstract
In this study, hydroxyapatite (denoted as HAp) nanostructure with uniform morphologies,
controllable size, nano-dispersion and narrow size distribution in diameter has been
synthesized successfully by low-temperature hydrothermal process, and the as-synthesized
powders were characterized by XRD, scanning electron microscopy, high-resolution
transmission microscopy, FT-IR, Zetasizer and inductively coupled plasma. In the present
work, a novel sonochemical technique using CaHPO4·2H2O/NaOH/distilled water with
cetyltrimethylammonium bromide ((CH3(CH2)15N+(CH3)3Br−) designated as CTAB) under a
hydrothermal condition to synthesize HAp nanostructure was described. Furthermore, the
usage of a high basic condition and a water environment are the two crucial keys in ensuring
the formation of HAp in the hydrothermal/sonochemical processes. However, the crystallite
size and crystallinity degree of the HAp increased with increasing annealing temperature.
Indeed, the present work will introduce a new method in synthesizing HAs for scientific and
medical engineering.
(Some figures in this article are in colour only in the electronic version)

1. Introduction dicalcium phosphate dihydrate (CaHPO4·2H2O, DCPD) and

hydroxyapatite with a hexagonal structure with space group
There is an escalating interest in calcium phosphates, P63/m and lattice parameters a = b = 9.4225 Å and c =
particularly apatites, which seems to be driven mainly by 6.8850 Å [17]. There are two formula units per unit cell
the requirements for the development, understanding and and the arrangement is typical of the calcium apatites and
manufacture of biomaterials [1]. Compounds with the can be viewed as consisting of unconnected, robust PO3− 4
apatite (Ca10(PO4)6(F, Cl, OH)2) structure have been widely tetrahedral with Ca2+ in the space between, and a chain of
studied due to their potential applications as biomaterials [2], OH− ions along the c-axis to balance the change (figure 1).
catalysts [3], ionic exchangers [4], oxide ion conductors and Monoclinic structures with four formula units per unit cell have
luminescent materials [5]. Many synthetic strategies have also been reported [18–22], where the monoclinic unit cell is
been developed to produce these nanostructures, such as the obtained from the hexagonal one by doubling the b lattice
solid-state reactions [6] and sol–gel method [7]. Recently, parameter and by having different arrangements of the anion
hydrothermal reactions have been used successfully as a chains. Difficulties in obtaining the stoichiometric compound,
non-traditional way of producing nanopowder, single crystals particularly with regard to the hexagonal phase of HAp, present
and nanostructure [8–16]. Depending on the supersaturation a problem. Following failed attempts to fix x-ray diffraction
level and the solution pH, a number of calcium phosphates data using the space group P63, the structure was found with
may be formed at ambient temperatures and pressure. In the space group P63/m through the addition of two mirror
an aqueous solution of pH > 4, the order of increasing planes perpendicular to [0 0 1] and passing through z = 1/4
solubility is as follows: tricalcium phosphate (Ca3(PO4)2, and z = 3/4. However, this structure has four possible sites
TCP), octa-calcium phosphate (Ca4H(PO4)3·2.5H2O, OCP), for the two OH groups per unit cell, a difficulty which was

1748-6041/08/025002+06$30.00 1 © 2008 IOP Publishing Ltd Printed in the UK

Biomed. Mater. 3 (2008) 025002 S A Manafi et al

experiment, a solution was first prepared by dissolving

Ca(NO3)2, (NH4)2HPO4 and distilled water in amounts
corresponding to the molar ratio of calcium to phosphorus of
1.67. Subsequently, some NaOH was added to the solution
to keep its pH around 10. Before being transferred to a
Teflon-lined autoclave, the solution mixture was pretreated
in an ultrasonic water bath for 30–40 min. The hydrothermal
syntheses were conducted at 150 ◦ C for 25 h in an electric oven.
After the reactions, white paste products (HAp nanorods)
were harvested by and decanted with distilled water. The
obtained HAp nanorods were characterized with scanning
electron microscopy, energy-dispersive x-ray spectroscopy
(SEM/EDX, XL30) and transmission electron microscopy.
Figure 1. Hydroxyapatite structure viewed along the c axis.
The size distribution and morphology of the samples were
analyzed by a field emission gun (FEG) transmission
resolved by assuming 0.5 occupancy per site resulting from electron microscope and selected area electron diffraction
disorder in every anion column or disorder from column to (TEM/SAED observation on a Philips CM200 transmission
column [23]. Among them, HAp, thermodynamically the most electron microscope operated at 200 kV. X-ray diffraction
stable calcium phosphate salt, has been extensively studied patterns were recorded in the angular range 2θ = 20–60◦ .
because it is the main inorganic compound of hard tissues in For these experiments, a Siemens diffractometer (30 kV
vertebrates. Although the precipitation of HAp is of particular and 25 mA) with the Kα1 radiation of copper (λ = 1.5406 Å),
importance in the biomineralization processes, very little is was used. The powder product was further investigated using
known about the crystal growth mechanism. In the present Fourier transform infrared (FTIR) spectroscopy in a Bruker-IR
work, the hydrothermal technique was developed for the spectrometer from 500 to 4000 cm−1 using the KBr technique
formation of HAp nanostructures. The nano-hydroxyapatite and operating in the transmittance mode. Particle size of
is interestingly crystalline with a high yield. These high- HAp has been monitored by a Malvern 3000 HSA Zetasizer
quality HAp nanorods represent a well-defined nanoscale operating at a wavelength of 633.0 nm at ambient temperature.
structure needed for both fundamental studies and clinical Finally, the crystallite size of the powder was evaluated from
applications. the peak broadening of XRD patterns based on Scherrer’s
formula as follows [24]:
2. Experimental procedures 0.9λ
D= , (1)
FWHM · cos θ
The synthesis of stoichiometric HAp nanopowders was where D is the crystallite size (nm), λ is the wavelength
performed in a 25 mL Teflon-lined stainless steel autoclave. of the monochromatic x-ray beam (λ = 0.154 056 nm for
All the reactants, Ca(NO3)2 (99% MERCK), (NH4)2HPO4 Cu Kα radiation), FWHM is the full width at half-maximum
(99% Alfa) and NaOH (96% Aldrich), were of reagent for the diffraction peak under consideration (rad) and θ is the
grade and used without further purification. In a typical diffraction angle (deg).

2500

2000 211

1500
Counts

112

1000
002

312 213

500 300 310

210 321 004

320
102

0
20 25 30 35 40 45 50 55 60
2 Theta

Figure 2. X-ray diffraction pattern of the hydroxyapatite.

2
Biomed. Mater. 3 (2008) 025002 S A Manafi et al

Size distribution (s)

40
% in class

30

20

10

(a)

5 10 50 100 500 1000

Diameter (nm)

Figure 3. The nanoparticle size of the hydroxyapatite product

measured by Zetasizer.

3. Results and discussion

The XRD patterns in figure 2 show that the as-synthesized

HAp sample at 150 ◦ C for 18 h is the hexagonal phase
with cell constants a = 9.22 Å and c = 6.885 Å, which
are very close to the values in the literature [25]. It is
interesting to note that HAp was obtained at such a low
(b)
temperature. According to recent research the temperature at
which a HAp nanocrystal changes from the amorphous phase Figure 4. SEM images of (a) as-synthesized HAp obtained under a
to the hexagonal phase when treated with the hydrothermal hydrothermal condition and (b) high aspect ratio nanorods.
method was up to 150 ◦ C, and a nearly complete hexagonal
phase was obtained at 150 ◦ C. The shape of the strong
diffraction peaks indicates that the samples are fairly highly
crystallized. The crystallinity of HAp powders synthesized
via a hydrothermal method was much higher than those
synthesized via normal chemical precipitation [26], sol–
gel [27] or normal micromulsion methods [28], which was
attributed to the hydrothermal treatment in the synthetic
process [29]. The XRD results showed that the bioceramics
were composed of highly crystalline and single-phase HAp,
and no obvious impurity phase could be found. The shape of
the diffraction peaks suggests that the sample could be well
crystallized. The broadened nature of these diffraction peaks
implies that the grain sizes of the sample are nanometer scale.
Estimating from the Debye–Scherrer formula, the average
grain size is 15 ± 5 nm.
The nanoparticle size of the HAp product was analyzed
using a Zetasizer method. These measurements reveal the Figure 5. HRTEM images of HAp nanorods.
particles to have a wide distribution (figure 3).
Figure 4(a) shows the SEM image of HAp nanostructures. nanostructures, because long-time ultrasonic treatment could
The typical spherical HAp nanostructures consisted of rod-like not destroy the nanostructures. SEM again reveals many
HAp 15–50 nm in diameter and several micrometers in length. samples to be made up of 1 µm dimension hexagonal-shaped
The definition of the rod-like HAp nanostructures comes from crystallites, as shown in figure 4(b).
the geometrical similarity to flowers. In our experiments, In figure 5, the TEM image reveals the morphology of
it is impossible that the aggregation resulted in the rod-like nanostructures formed at 150 ◦ C for 25 h with diameter

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Biomed. Mater. 3 (2008) 025002 S A Manafi et al

Figure 6. SAED of the hydroxyapatite nanostructure.

Figure 7. Energy dispersive x-ray analysis (EDAX) of the obtained hydroxyapatite products.

15–40 nm and length 70–150 nm. It is quite obvious that values are calculated from Bragg’s diffraction equation using
the morphologies of the products change considerably as a the diffraction ring diameter and the camera length of the TEM.
function of the residence time in the autoclave. On the basis The chemical stoichiometry of nanorods was investigated
of the morphologies observed by TEM, it can be concluded with EDX (figure 7) which indeed gave an atomic ratio of
that the size of the particles increases with increasing reaction HAp ∼1.67. At the same time, this result is consistent with
time, a fact that is consistent with the results of the XRD and the inductively coupled plasma (ICP) calculation. More details
Zetasizer. about the structure of HAp nanostructures were investigated
by the SAED pattern and high-resolution transmission
The selected area electron diffraction (SAED) pattern
microscopy (HRTEM).
(figure 6) taken from the as-prepared HAp nanostructure
Fourier transform infrared (FTIR) analysis revealed the
synthesized at 150 ◦ C for 25 h consists of a number of presence of carbonate on the surface of the HAp. Figure 8
rectangular and some distinct spots along the ring contours, shows the transmittance infrared spectrum of synthetic HAP
suggesting a hexagonal structure. The spots in an electron in the 4000–650 cm−1 region. The two sharp and weak bands
diffraction pattern arise due to the diffracted electron beam at 2824 cm−1 are attributed to a small amount of residual
from a set of lattice planes in the crystallites present in the CTAB, while a narrow band located near 965 cm−1 (962 cm−1
sample satisfying the Bragg diffraction condition. In other in figure 8) represents the υ 1 mode of PO3− 4 ions in apatite.
words, the ring is an envelope of all diffracted spots. Among The main signal of phosphate appears in the triply degenerate
the same rings a few spots appear to be prominent, which υ 3 domain (1000–1100 cm−1). The adsorption band at
indicates the formation of crystallites. The interplanar spacing 3500 cm−1 confirmed the presence of OH− groups. The υ 2

4
Biomed. Mater. 3 (2008) 025002 S A Manafi et al

1.4

1.2

Transmittance (%)
1

470

870
0.8
1614
1400
0.6

3570
0.4 965 3400-3500

0.2 609
559

1000-1100

0
100 600 1100 1600 2100 2600 3100 3600 4100
-1
Wavenumber (cm )
Figure 8. The FTIR spectrum of the HAp nanostructures.

−1
peak of CO2−3 is located at 875 cm ; this absorption results
[4] Ando J and Matsuno S 1968 Bull. Chem. Soc. Japan 41 342
[5] Blasse G 1987 Mater. Chem. Phys. 16 201
from out-of-plane stretching. The υ 3 mode, near 1400 cm−1,
[6] Nakayama S and Sakamoto M 1998 J. Eur. Ceram.
is the strongest IR peak for carbonate. This peak is actually Soc. 18 1413
composed of two bands (1421 cm−1, in figure 8) [30, 31]. The [7] Tao S and Irvine J T S 2001 Mater. Res. Bull. 36 1245
shape of the υ 3 signal and the absence of the C–O absorption [8] Xia Y, Yang P, Sun Y, Wu Y, Mayers B, Gates B, Yin Y,
bands at 700 cm−1 indicate that no calcite was associated Kim F and Yan H 2003 Adv. Mater. 15 353
with the HAp. Carbonate ions can substitute for either OH− [9] Patel N, Gibson I R, Ke S, Best S M and Bonfield W 2001
J. Mater. Sci., Mater. Med. 12 181
or PO3− 2−
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