Materials Characterization 47 (2001) 83 – 87

Hydrothermal conversion of coral into hydroxyapatite

Ye Xu, Dazhi Wang*, Lan Yang,
Honggao Tang
Department of Materials Science and Engineering, University of Science and Technology of China, Hefei, Anhui, 230026,
People’s Republic of China
Received 21 November 2000; accepted 31 May 2001

Abstract

Porous coralline hydroxyapatite (CHA) was prepared by the hydrothermal conversion of Porites skeleton
(aragonite) with and without the use of the mineralizer KH2PO4. X-ray diffraction (XRD) analysis, Fourier
transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) were employed to characterize
the exchange process. The complete conversion was achieved after 10 and 17 days, respectively, with and without
the mineralizer. When the mineralizer was present, the exchange process involved only the direct conversion of
aragonite into HA. Without the presence of the mineralizer, there were, in addition, intermediary conversions of
aragonite into calcite and calcite into b-tricalcium phosphate (b-TCP). The research study also showed that the
presence of the mineralizer accelerated the conversion process and preserved the interconnecting microstructure of
the coral better. D 2001 Elsevier Science Inc. All rights reserved.

Keywords: Hydroxyapatite; XRD

1. Introduction achieved at elevated temperatures and pressures,

Hydroxyapatite [Ca10(PO4)6(OH)2; HA] has been
widely investigated as a bone substitute, because it
has been shown to form a direct bond with adjacent
hard tissue [1 – 6]. Current methods of preparing HA
range from an aqueous solution [7 – 9], solid-state
reaction [10], to hydrothermal methods [11]. Cor-
alline HA (CHA), prepared by the hydrothermal
conversion of marine coral, has been used extensively
in clinical applications in recent years [12]. Coral
exists in very large quantities and has a cancellous
pore structure essential for the in-growth of bone
tissue [13]. The hydrothermal conversion to HA is
insuring more than adequate sterilization and preserv-
ing the original structure of coral. However, the
complicated exchange process has not yet been
investigated in any detail. In this paper, we report
on the conversion of the calcium carbonate (aragon-
ite) skeleton of the marine coral Porites into HA, both
with and without the use of the mineralizer KH2PO4,
and the subsequent study of the exchange process.
2. Experimental procedure
2.1. Sample preparation

Porous CHA with micrometer-range intercon-

* Corresponding author. Tel.: +86-551-36011701. nected pores was prepared by mixing the Porites
E-mail address: dzwang@ustc.edu.cn (D. Wang). and (NH4)2HPO4, with or without the mineralizer

1044-5803/01/$ – see front matter D 2001 Elsevier Science Inc. All rights reserved.
PII: S 1 0 4 4 - 5 8 0 3 ( 0 1 ) 0 0 1 5 4 - 1
84 Y. Xu et al. / Materials Characterization 47 (2001) 83–87

KH2PO4, through the following procedure. First,

coral blocks obtained from the coast were cleaned,
dried and then sliced into average pieces (10
10

5 mm). The pieces of Porites coral together with
equal quantities of phosphate and water were placed
in an autoclave, occupying three-quarters of the
autoclave volume. The autoclave was then sealed,
heated at a specified temperature and pressure for
periods of time varying from 2 to 17 days, cooled,
and the products collected. In another autoclave,
under the same reaction conditions, a small quantity
of KH2PO4 was added as the mineralizer. The
following main exchange reaction took place:

10CaCO3 þ 6ðNH4 Þ2 HPO4 þ 2H2 O

) Ca10 ðPO4 Þ6 ðOHÞ2 þ 6ðNH4 Þ2 CO3 þ 4H2 CO3

2.2. Sample characterization

The phases of the products were studied using X-

ray diffraction (XRD) analysis (Kingaku D/Maz-g),
Fourier transform infrared (FTIR) spectroscopy
(using a Bruker Vector-22 spectrometer) and a Hita-
chi X-650 scanning electron microscope (SEM). The
Fig. 2. XRD spectra of CHA produced with mineralizer
KH2PO4 present for (a) 2, (b) 6, (c) 8, and (d) 10 days.

microstructural observations in the SEM were per-

formed on gold-coated specimens.

3. Results and discussions

3.1. XRD analysis

Figs. 1 and 2 show the XRD patterns obtained

Fig. 1. XRD spectra of CHA produced without mineralizer
present for (a) 2, (b) 6, (c) 8, (d) 10, (e) 13, and (f) 17 days.
from the hydrothermal conversion products at various
stages during the conversion process, both without
and with the mineralizer. Fig. 1a, the XRD pattern of
the CHA produced after 2 days with no mineralizer
present, shows a well-crystallized pattern of HA with
sharp peaks (Fig. 1a). The original aragonite phase is
also detectable. When the reaction time was extended
to 6 days (Fig. 1b), more HA was produced and less
aragonite remained. After 8 days (Fig. 1c), the
aragonite diffraction lines have disappeared com-
pletely and new diffraction lines are seen at 2q values
of 31.1, 34.1 and 27.8. These reflections are
attributed to the formation of b-tricalcium phosphate
(b-TCP). Calcite diffraction lines were also detected
at this stage. The XRD patterns obtained after 10
(Fig. 1d) and 13 days (Fig. 1e) show the gradual
disappearance of b-TCP and its replacement by HA.
After 17 days, only HA was detected (Fig. 1f). From
 Y. Xu et al. / Materials Characterization 47 (2001) 83–87 85

these observations, a complex transformation process

is inferred. This involves parallel reactions, one in
which the aragonite is converted directly into HA and
a second in which there is an intermediate trans-
formation of aragonite into calcite, a more thermo-
dynamically stable form of calcium carbonate [14]. In
the early stages of the conversion, aragonite trans-
formed to HA faster than to calcite, so the main
product detected was HA. As the change from
aragonite to calcite being slower than that to HA,
the calcite phase diffraction lines began to appear
later in time. The calcite was subsequently trans-
formed into b-TCP because of their structural sim-
ilarity [15]. However, HA is more thermodynamically
stable than b-TCP [16], so after 17 days, only pure
HA was produced.
By contrast, no conversion of aragonite into
calcite and formation of b-TCP were observed in
the experiment incorporating the mineralizer KH2PO4
(Fig. 2), all the aragonite being converted directly
into HA. From this, it was inferred that the mineral-
izer KH2PO4 accelerated the direct conversion of

Fig. 4. IR spectra of CHA produced with mineralizer

KH2PO4 present for (a) 2, (b) 6, (c) 8, and (d) 10 days.

aragonite into HA and inhibited the conversion of

aragonite into calcite.

3.2. FTIR spectroscopy

FTIR spectroscopy furnished additional informa-

Fig. 3. IR spectra of CHA produced without mineralizer
present for (a) 2, (b) 6, (c) 8, (d) 10, (e) 13, and (f) 17 days.
tion of the structure of CHA. Fig. 3 illustrates the
spectra of CHA formed without the mineralizer being
present. The complex changes detected in the XRD
spectra were also found in the FTIR spectra. The IR
absorption spectrum of the CHA was characterized by
three main regions, which are related to the vibra-
tional frequencies of the CO32  , OH  and PO43 
ions [17]. The peaks seen at 712.4 and 699.0 cm  1
in Fig. 3a and b were due to CO32  in aragonite,
while that seen at 712.2 cm  1 in Fig. 3c – e was due
to CO32  in calcite [18]. This confirmed the change
of aragonite to calcite observed in XRD patterns.
When the reaction time was increased, these carbon-
ate bands became less intense then disappeared
completely (Fig. 3f). In addition, there are CO32 
bands at 1422, 1453 and 1548 cm  1 in Fig. 3f. This
suggests that a small amount of carbonate remained
and took up the partial positions of OH  and PO43 
in the HA lattice. Types A and B carbonated HA were
produced, as revealed by the CO32  bands at 1548
and 1460 cm  1, which correspond to Type A carbo-
nated HA, and by those at 1460 and 1415 cm  1,
86 Y. Xu et al. / Materials Characterization 47 (2001) 83–87

which correspond to Type B carbonated HA [19]. At
the same time, the bands that corresponded to the
stretching (3565 cm  1) and librational (633 cm  1)
modes of the OH  groups became more intense with
duration, which means that OH  groups had been
gradually incorporated into the HA lattice [17,20].
The FTIR spectra obtained from CHA formed in
the presence of the mineralizer are shown in Fig. 4
and it is apparent that there are some differences from
the spectra of Fig. 3. At the beginning of the
exchange process, the peak at 855 cm  1 is very
strong, which suggests that CO32  mostly occupied
the PO43  position in the HA lattice and thus
represents Type B carbonated HA. This peak gradu-
ally weakened as the exposure time increased, so that
after 10 days, at the end of the reaction, it had almost
disappeared. On the other hand, the peak at 877
cm  1, representing Type A carbonated HA, strength-
ened with time. This indicates that the Type B form
gradually transformed to Type A HA. The mech-
anism of this change is presently under study.
3.3. Scanning electron microscopy
Fig. 5a is a scanning electron micrograph of the
original Porites. The skeleton is permeated by inter-
connecting channels forming a fenestrate microstruc-
ture that has the advantage of allowing circulation of
body fluids and of increasing the potential for firm
attachment of body tissue. Fig. 5b and c show the

Fig. 5. Scanning electron micrographs of (a) Porites, (b) CHA completely exchanged without mineralizer present, and (c) CHA
completely exchanged with mineralizer KH2PO4 present.
 Y. Xu et al. / Materials Characterization 47 (2001) 83–87
structures of completely exchanged CHA without and
with mineralizer, respectively. In both cases, the porous
interconnecting structure of the Porites was preserved
but the size of pores has increased a little. In the
exchange process without mineralizer, the pores sub-
sided gradually with time. However, the metamorph-
osis of the pores was much smaller in the products
formed with mineralizer present. It appears that the
microstructure of the HA formed in the presence of the
mineralizer KH2PO4 was retained better than that
obtained without the mineralizer present.
4. Conclusions
The present study has revealed the complicated
processes involved in the hydrothermal conversion
of coral to HA. The conversions of aragonite into
calcite, of calcite into b-TCP and of b-TCP into
HA were found when the hydrothermal reaction
was conducted without the presence of a mineral-
izer. However, when the hydrothermal conversion
was performed with the mineralizer KH2PO4 pre-
sent, only the conversions of aragonite into HA and
Type B carbonated HA into Type A carbonated HA
were detected. It was found the mineralizer
KH2PO4 used in the exchange process simplified
the conversion, increased the conversion rate and
retained better the original interconnecting structure
of coral.
References
[1] Dennisen HW, de Groot K, Makkes PC, Hoff A, van
den Klopper PJ. Tissue response to dense apatite im-
plants in rats. J Biomed Mater Res 1980;14:713 – 21.
[2] Galgut PN, Waite IM, Tinkler SMB. Histological in-
vestigation of the tissue response to hydroxyapatite
used as an implant material in periodontal treatment.
Clin Mater 1990;6:105 – 21.
[3] Jianguo Z, Xingdong Z, Muller-Mai C, Gross U. The
early host and material response of hydroxyapatite/b-
tricalciumphosphate porous ceramics after implanta-
tion into the femur of rats. J Mater Sci: Mater Med
1994;5:243 – 51.
[4] Osborn JF, Kovacs E, Kallenberger A. Hydroxyapatite
87
— development of a new biomaterial and first exper-
imental results. Dtsch Zahnaerztl Z 1980;35:54 – 6.
[5] Reference deleted in proof.
[6] Furlong RJ, Osborn JF. Fixation of hip prostheses by
hydroxyapatite ceramic coating. J Bone Jt Surg 1991;
73-B:741 – 5.
[7] Arends J, Christoffersen J, Christoffersen MR, Eck-
ert H, Fowler BO, Heughebaert JC, Nancollas JH,
Yesinowski JP, Zawacki SJ. A calcium hydroxyapa-
tite precipitated from an aqueous solution: an interna-
tional multimethod analysis. J Cryst Growth 1987;84:
515 – 32.
[8] Osaka A, Miura Y, Takeuchi K, Asada M, Takahashi
K. Calcium apatite prepared from calcium hydroxide
and orthophosphoric acid. J Mater Sci: Mater Med
1991;2:51 – 5.
[9] Slosarczak A, Stobierska E, Paszkiewicz Z, Gaw-
licki M. Calcium phosphate materials prepared from
precipitates with various calcium: phosphorus molar
ratios. J Am Ceram Soc 1996;79:2539 – 44.
[10] Young RA, Holcomb DW. Variability of hydroxyapa-
tite preparations. Calcif Tissue Int 1982;34:17 – 32.
[11] Yoshimura M, Suda H, Okamoto K, Ioku K. Hydro-
thermal synthesis of biocompatible whiskers. J Mater
Sci 1994;29:3399 – 402.
[12] Suchanek W, Yoshimura M. Processing and properties
of hydroxyapatite-based biomaterials for use as hard
tissue replacement implants. J Mater Res 1998;13:
94 – 117.
[13] Roy DM, Linnehan SK. Hydroxyapatite formed from
coral skeletal carbonate by hydrothermal exchange.
Nature 1974;246:220 – 2.
[14] Holmes R, Mooney V, Bucholz R. A coralline hydrox-
yapatite bone graft substitute preliminary report. Clin
Orthop 1984;188:252 – 62.
[15] Lippmann F. Sedimentary carbonate minerals. New
York: Springer-Verlag, 1973. p. 97.
[16] Brown WE, Chow LC. Cements research progress.
Westerville, OH: American Ceramic Society, 1986.
p. 352.
[17] Vaidya SN, Karunakaran C, Pande BM, Gupta NM.
Pressure-induced crystalline to amorphous transition
in hydroxyapatite. J Mater Sci 1997;32:3213 – 7.
[18] Andersen FA, Brecevic L. Infrared spectra of amor-
phous and crystalline calcium carbonate. Acta Chem
Scand 1991;45:1018 – 24.
[19] Roy DM, Eysel W, Dinger D. Hydrothermal synthesis
of various carbonate-containing calcium hydroxyapa-
tite. Mater Res Bull 1974;9:35 – 40.
[20] Cao Y, Weng J, Cheng J, Zhang X. Water-vapor-treated
hydroxyapatite coatings after plasma-spraying and
their characteristics. Biomaterials 1996;17:419 – 24.