Colloids and Surfaces A: Physicochem. Eng.

Aspects 296 (2007) 149–153

Synthesis of hydroxyapatite nanoparticles using an

emulsion liquid membrane system
Somnuk Jarudilokkul a,∗ , Wiwut Tanthapanichakoon b , Virote Boonamnuayvittaya a
a Department of Chemical Engineering, King Mongkut’s University of Technology, Thonburi, Bangkok 10140, Thailand
b Department of Chemical Engineering, Chulalongkorn University, Pattumwan, Bangkok 10330, Thailand

Received 30 May 2006; received in revised form 19 September 2006; accepted 22 September 2006
Available online 28 September 2006

Abstract
Hydroxyapatite (HAp) has been synthesized by various methods, such as co-precipitation, sol–gel, and microemulsion. However, particle size,
particle size distribution and cost were drawbacks of those techniques. In this study, HAp nanoparticles were prepared by means of emulsion liquid
membrane system (ELM) or water-in-oil-in-water system (W/O/W). The aim of the work was to evaluate the preparation of HAp nanoparticles
using ELM consisting of Span 20 and Tween 80 as a mixture of biodegradable surfactant and caproic acid as an extractant. The effect of Ca/P ratio,
reaction temperature, and calcination temperature on the characteristics of synthesized particles was investigated. The synthesized HAp particles
were amorphous calcium deficient apatites. Ca/P molar ratio of the synthesized particle (Ca/Pp ) increased with increasing initial Ca/P molar ratio
(Ca/Pi ) and reach maximum at Ca/Pp of 1.65. We found significant amount of ␤-TCP in the particles prepared with high initial Ca/Pi molar ratio
of 2.00 at which the undesired reactions at high Ca2+ concentration occurred. Increasing of reaction and calcinations temperature resulted in the
reduction of surface area. After thermal treatment at temperatures ranged from 450 to 750 ◦ C, the specific surface area reduced from 227 to 58 m2 /g.
At temperature of 750 ◦ C, we found that the morphology of microsized particles instead of nanosized ones.
© 2006 Elsevier B.V. All rights reserved.
Keywords: Hydroxyapatite; Emulsion liquid membrane; Nanoparticle; Biodegradable surfactant; Characterization

1. Introduction tors, and recently bioceramics but such technique is not the cost
effective one.
Hydroxyapatite (HAp), with the structural formula of An emulsion liquid membrane (ELM, water-in-oil-in-water
Ca10 (PO4 )6 (OH)2 , is the major constituent of human bones and (W/O/W) emulsion) system has been studied for the selective
teeth. Synthetic HAp is now widely used in medical applica- separation of heavy metals, in which the metal ions in the exter-
tions, such as implants or as coatings on prostheses [1,2] and nal water phase are extracted into the organic membrane phase,
used in Michael addition reaction as the basic catalyst [3]. and then stripped and concentrated into the internal water phase.
Several chemistry-based processing routes have been reported Recently, it has been found that the internal water phase is capa-
for preparing fine and agglomerate-free ceramic hydroxyapatite ble of being used for the preparation of size-controlled and
powders, although the degree of success varies considerably morphology-controlled fine particles, since the micron-sized
from one route to another. Moreover, other synthesis tech- internal water droplet has a restricted reaction area [5,6]. For
niques are co-precipitation technique, sol-gel processing route, such reason, ELM technique is a promising method for the
pyrolysis of aerosols, hydrothermal processing routes, post- preparation of HAp fine particles with narrow size distribution
reaction refinement by emulsion, inverse microemulsion route, range.
and microemulsion processing route [4]. Microemulsion pro- In this present work, the ELM system was prepared from
cessing route has been used successfully to prepare nanoparticles biodegradable surfactant and fatty acid as an extractant. It was
of metals, oxides, halides, sulfides, carbonates, superconduc- performed in order to prepare nanosized hydroxyapatite particle.
The effect of initial Ca/P molar ratio in the feed solution, reaction
temperature and calcination temperature were investigated. The
∗ Corresponding author. Tel.: +66 2470 9221 30x221; fax: +66 2428 3534. textural characteristics and morphology of HAp particles were
E-mail address: [email protected] (S. Jarudilokkul). finally examined by XRD and SEM.

0927-7757/$ – see front matter © 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.colsurfa.2006.09.038
150 S. Jarudilokkul et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 296 (2007) 149–153

2. Materials and methods

The chemicals used in this work include CaCl2 ·2H2 O,

(NH4 )2 HPO4 , iso-octane and ammonia solution (25%) which
were used as a Ca2+ source, a PO4 3− source, a OH− source and
an organic membrane phase, respectively. They were AR grade
and obtained from Merck. The biodegradable nonionic surfac-
tants, Span 20 and Tween 80, were supplied by Fluka. Fatty
acid, n-caproic acid was used as an extractant (carboxylic acid
extractor) was supplied by Sigma.

2.1. Particle synthesis

An internal water phase of W/O emulsion was 30 mL of 0.5 M

(NH4 )2 HPO4 solution solubilized in the organic membrane
phase at volume ratio of 3:4. The membrane phase comprises
iso-octane, n-caproic acid (1.5 mL) and 5 g of surfactant mixture Fig. 1. Scanning electron micrographs for particles prepared at Ca/Pi = 1.67 in
(1:1) of Span20 and Tween80. They were emulsified mechani- ELM system.
cally by mixing with a homogenizer at the rotation of 14,000 rpm
for 5 min. The obtained W/O emulsion was added to an exter- the preparation of size-morphology-controlled fine particles.
nal water phase containing 1.0 M CaCl2 ·2H2 O (15–30 mL) and The SEM and TEM images of the particles obtained at the ini-
50 mL of 25% ammonia solution. The total volume of external tial Ca/Pi molar ratio of 1.67 in the ELM system are shown in
water phase volume was controlled at constant volume of 80 mL Figs. 1 and 2, respectively. The ELM systems yielded nanoparti-
by adding deionized water. cles (smaller than 70 nm) with spherical morphology and narrow
Stirring at vigorously speed was required to disperse W/O particle size range which are significantly different from those
emulsion droplet to form W/O/W emulsion. After stirring for that prepared by precipitation method. In precipitation tech-
30 min, the W/O/W emulsion was left to allow aging and sepa- nique, the calcium solution was directly mixed with phosphate
rating the W/O emulsion from external water phase. The upper solution under same condition and reaction time, the larger size
emulsion phase was washed with deionized water, followed particles with irregular shapes were obtained.
by centrifugation (12,000 rpm 30 min at 25 ◦ C) to remove both The nanometer particle size was confirmed by measurement
residue external phase and organic membrane phase. Then, the of emulsion droplet size in external water phase. Fig. 3 shows the
solid particles were washed with ethanol followed by centrifuga- emulsion droplet size evolution within 0–60 min. The emulsion
tion in order to completely remove the organic membrane phase. droplet size distributions were in the range of 1–100 ␮m. Within
The particles obtained were finally dried at 105 ◦ C for 2 h. The 30 min, about 60% of emulsion, the droplet size was lower than
effects of initial Ca/P molar ratio (Ca/Pi ), reaction temperature 10 ␮m and no significant variation in size distribution can be
and calcination temperature in the range of 1.00–2.00, 30–80 identified. The increase of emulsion droplet size was observed
and 450–750 ◦ C were studied, respectively. after 1 h of reaction. In general, the size of internal water droplets
inside the emulsion phase should be comparatively smaller than
2.2. Characterization that of the emulsion droplets in the external water phase. There-

The synthesized particles were characterized by means of

scanning electron microscopy (SEM), JSM631F, transmission
electron microscopy (TEM), TEOL 2010, powder X-ray diffrac-
tion (XRD), (40 kV-30 mA, Cu K␣, Rigaku). A Laser Scattering
Particles Size Distribution Analyzer, Malvern Master Sizer
2000, was used for measuring the size of W/O emulsion droplet
size. The molar ratio Ca/Pp in the synthesized particles was
determined using an inductive coupled plasma atomic emission
spectrometer (ICP-AES 8440 Plasmalab).

3. Results and discussion

3.1. HAp particles characteristics

In ELM system, the cationic species are extracted from an

external water phase into an internal water phase and then formed
particles. The internal water phase, therefore, is being used for Fig. 2. TEM micrographs of HAp particles prepared by ELM technique.
 S. Jarudilokkul et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 296 (2007) 149–153 151

Table 1
The BET surface area of HAp particle at different feed concentration

Ca/Pi A (m2 /g)a VBJH (cm3 /g)b Dpore (nm)c

1.00 180.2 0.7625 15.7

1.33 198.5 0.8344 15.9
1.67 227.2 0.8225 16.1
2.00 181.6 0.7611 16.2
a External surface area by BET method.
b BJH method cumulative pore volume.
c BJH method pore diameter.

The relationship between calcium to phosphate initial molar

ratio or Ca/Pi and Ca/Pp in the synthesized particles is shown in
Fig. 5. The Ca/Pp molar ratio increased with increasing Ca/Pi
and reached the maximum at the Ca/Pp around 1.65. Then, Ca/Pp
Fig. 3. Time-course variation for an emulsion droplet size distribution within declined to 1.60 when the Ca/Pi was around 2.00. We considered
the reaction time. that the HAp particles prepared by ELM technique with Ca/Pi
ratio from 1.60 to 2.00 should be calcium deficient apatites.
fore, the data confirms that particles obtained in the ELM system The same trend was also reported in the preparation of calcium
should be smaller than the internal water droplet size. phosphate particles using ELM consisting sorbitan sesquioleate
The XRD patterns of the particles prepared by ELM system and 2-methyl-2-ethyl heptanoic acid [5,6].
and simple precipitation are shown in Fig. 4. It was verified The XRD patterns of obtained HAp particles prepared at dif-
that the particles obtained by the ELM system were amor- ferent Ca/Pi ratio are shown in Fig. 6. It was found that the
phous hydroxyapatites whereas the morphology of the particle synthesized HAp particles at various Ca/Pi ratio were low crys-
prepared by simple precipitation technique was relatively crys- talline. At the highest Ca/Pi ratio of 2.00, the XRD pattern of
talline. HAp shows board and lowest intensity peak that implies the
relatively low in crystallinity or they are amorphous materials.
3.2. Effect of initial Ca/Pi molar ratio Fig. 7 presents the XRD pattern of HAp particles after calci-
nation at temperature of 650 ◦ C. It was found that the preparation
Table 1 shows the effect of Ca/Pi molar ratio on the BET sur-
of HAp particles at high Ca/Pi molar ration of 2.00 induced
face area of the particles. The BET surface area of HAp particles
the significant amount of ␤-tricalcium phosphate (␤-TCP). We
prepared were in the range of 180–227 m2 /g. We found that the
consider that at high Ca/Pi ratio, the transport rate of Ca2+ is
HAp particles had very high surface area compared to those
comparatively higher than those of low Ca/Pi ratio, therefore
prepared by simple precipitation technique which was in the
the excess amount of Ca2+ ions in the internal water phase react
range of 30–108 m2 /g [2,4,7]. Since the surface area of particles
with PO4 3− within internal water phase and possibly form oth-
directly related to its particles size therefore, we can estimate the
ers type of calcium phosphates. Hirai et al. [5,6] reported that
particles size to be around 8–10 nm based on the BET surface
the value of Ca/P ratio of ␤-TCP was 1.50. Therefore, the prepa-
area. We conclude that the variation of initial Ca/P molar ratio
ration of HAp particles at Ca/Pi of 2.00 had Ca/Pp lower than
had almost no effect on the sizes of formed particles.
1.67 which indicated the present of ␤-TCP.

Fig. 4. X-ray diffraction patterns for particles prepared at Ca/Pi = 1.67 in ELM
system and simple precipitation technique (a) ELM system (b) simple precipi- Fig. 5. Relationship between the Ca/P molar ratio for hydroxyapatite prepared
tation. using the ELM technique, Ca/Pp , and the initial molar ratio, Ca/Pi .
152 S. Jarudilokkul et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 296 (2007) 149–153

Fig. 8. Specific surface area of apatite obtained at different temperatures.

Fig. 6. The X-ray diffraction patterns of hydroxyapatite particles prepared at
different Ca/Pi molar ratio using ELM technique.
of organic membrane phase, which was resemble to simple pre-
cipitation phenomenon. In general, the size of particles obtained
3.3. Influence of the reaction temperature (Tr ) by precipitation synthesis was found in the range of microsized
particles.
When the reaction temperature was increased from 30 to 50 In addition, the degree of supersaturation increased when
and 80 ◦ C, as presented in Fig. 8, it was found that the BET increasing temperature resulted in the solubility constant (Ksp )
surface area of the HAp particles with Ca/Pi of 1.67 exponen- of HAp decreased [9]. The formation of HAp particle from such
tially decreased from 227 to 147 and 98 m2 /g, respectively. At condition produced more crystallinity HAp particles. Therefore,
temperature of 80 ◦ C, the stability of emulsion was low result- the XRD patterns of HAp particles demonstrate the narrower
ing in the negatively charge ions transport out of the internal pattern, as shown in Fig. 9.
water phase. Therefore, the BET surface area of HAp particles
prepared by emulsion method was very close to that obtained 3.4. Influence of the calcinations temperature (Tc )
from simple precipitation technique [7]. Moreover, the reac-
tion temperature affects the emulsion stability and the transport The objective of calcination was to eliminate undesired
rate of ions. When reaction temperature increases, the emulsion extraneous compounds in the materials. However, excessive
stability should decrease and finally the emulsion will break [8]. calcination temperature will reduce the surface area of the mate-
For the transport rate of ions, the positively charge ions should rials. As we know that the nanosized particles are very sensitive
transport with relatively low velocity. Therefore, increasing tem- to heat, hence the sintering of nanosized particles probably occur
perature may cause more negatively charge ions transport out during the calcination. Fig. 10 shows the effect of calcination
of the internal water phase. Subsequently, the reaction to form temperature on BET surface area. It was found that the sur-
HAp particles should occur in bulk solution or external surface face area decreased with increasing calcination temperature.
In addition, the micro-sized particles were obtained instead of

Fig. 7. The X-ray diffraction pattern of hydroxyapatite prepared using ELM

technique after calcinations at 650 ◦ C. Fig. 9. XRD patterns of apatite synthesized at various reaction temperatures.
 S. Jarudilokkul et al. / Colloids and Surfaces A: Physicochem. Eng. Aspects 296 (2007) 149–153 153

amorphous phase and occurs at 635 ◦ C [2]. Thus, when the calci-
nation temperature was higher than crystallization temperature,
the impurity as ␤-TCP was observed as shown in Fig. 11.

4. Conclusions

The internal water phase of emulsion liquid membrane was

able to use for the preparation of size and morphology-controlled
fine particles. In this work, the preparation of nanosized calcium
phosphate particle was performed. The synthesized particles
were calcium deficient apatite in amorphous morphology. The
variation of initial Ca/P molar ratio had no effects on the parti-
cle sizes. However, preparation with high Ca/P ratio caused the
formation of ␤-TCP in the product. By increasing the reaction
temperature, the surface area of prepared particles decreased
Fig. 10. Specific surface area of HAp particle after calcination at various calci- which was related to the increasing in particles size. The refor-
nation temperatures. mation of HAp particles to ␤-TCP was also observed during the
calcinations.

Acknowledgement

This work received partial financial support from Thailand

Research Fund (TRF-RTA of Dr. Wiwut Tanthapanichakoon
2002-2005).

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