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Article history: The present work reports the successful synthesis of nano-hydroxyapatite, Ca10 (PO4 )6 (OH)2 (denoted
Received 5 January 2009 HAP) from calcined eggshell by hydrothermal method using cationic surfactant (CTAB) as regulator of
Received in revised form 13 August 2009 nucleation and crystal growth. The reaction involved in the synthesis was studied elaborately. The influ-
Accepted 12 September 2009
ence of reaction temperature, ageing time and CTAB concentration on the synthesis of nano-HAP are also
studied in addition to the effect of sintering temperature on the crystal growth. Spectral characterization
Keywords:
involving Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) techniques were
Nano-hydroxyapatite
performed for functional group analysis and phase identification of the materials, respectively. Thermal
Eggshell
Cationic surfactant
stability of nano-HAP was investigated by thermal analysis (TG/DTA). The physical characteristics, such
Hydrothermal method as morphology and particle size of the synthesized nano-HAP were assessed thoroughly by scanning
electron microscopy (SEM) and atomic force microscopy (AFM) techniques. The results have revealed
that well-crystallized nano-HAP was synthesized by hydrothermal treatment at 160 ◦ C for 10 h with the
addition of CTAB at critical micelle concentration (CMC). It was also found that the synthesized nano-HAP
was thermally stable up to 1100 ◦ C.
© 2009 Elsevier B.V. All rights reserved.
1386-1425/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.saa.2009.09.021
1128 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134
vided there is a degree of complementarity in charge and size sample) was determined by EDAX. Atomic force microscopy (AFM)
between the guest ions and interstices in the structure of the to study the surface topography of the samples was performed
crystal boundary layers. However, the samples prepared at ambi- using a Picoscan 2100 (Molecular Imaging, USA) employing Au
ent temperature were generally poorly crystallized so that their coated silicon nitride pyramidal probes with tip size of ∼30 nm.
strength was not high. Therefore, it is believed that the combina- The as-prepared samples were ground into a fine powder and were
tion of the two methods might give a better result. In the present uniaxially pressed using a stainless steel die at 125 MPa to form
study, we explore the assistance of surfactant in HAP preparation a pellet of size 10 mm × 0.3 mm diameter and the pressed bodies
by hydrothermal method from eggshells, a waste material. Calcined were sintered at 900 ◦ C for 2 h for analyzing the surface topogra-
eggshell and phosphoric acid were used as calcium and phospho- phy using AFM. The weight loss and thermal stability of the samples
rous sources, respectively. A cationic surfactant, cetyltrimethyl or were also evaluated from the thermogravimetric analysis data. An
hexadecyltrimethylammonium bromide (CTAB), which is widely HETZSCH-Geratebau GmBH thermal analyser having STA (simulta-
used to achieve control of morphology in many aqueous synthetic neous thermal analyzer) 409C microbalance with a heating rate of
methods, was chosen as the regulator to control the crystalliza- 10 ◦ C min−1 was employed.
tion of HAP. The influence of reaction temperature, ageing time,
CTAB concentration and sintering temperature on the nucleation 3. Results and discussion
and crystal growth process is investigated.
3.1. Synthesis of HAP
2. Materials and methods
The present study employs hydrothermal method for the pro-
duction of HAP using calcined eggshell as calcium source and
2.1. Powder preparation
phosphoric acid as phosphorous source. The thermal treatment
used for eliminating the organic component of eggshell at 900 ◦ C
The surface area of the eggshells collected were mechanically
(1 h) converts calcite (CaCO3 ) into calcium oxide by freeing carbon
cleaned using double distilled water and were dried in an air oven
dioxide (CO2 ). The expected reaction is
for 1 h at 100 ◦ C. The dried eggshells were ground into fine powder
and calcined at 900 ◦ C for 1 h. The amount of calcium present in CaCO3 → CaO + CO2 ↑ (1)
calcined eggshell was estimated by conventional EDTA titrimetric
method. A stoichiometric amount of the calcined eggshell powder Fig. 1 shows the FT-IR spectra of raw, 800 and 900 ◦ C cal-
was dispersed in doubly distilled water (pH∼12). To the phospho- cined eggshells, respectively. The FT-IR spectra shown in Fig. 1a
ric acid solution (0.075 M), different concentrations of CTAB was and b indicate characteristics peaks corresponding to CO3 2− ion at
added and kept overnight (about 8 h) to ensure the co-operative 1420–1570 and 875 cm−1 . The carbonate ion peak for 800 ◦ C cal-
interaction and self-assembly process was completed. The phos- cined eggshell was found to be intense when compared to the raw
phate solution was then added drop wise to calcium suspension at eggshell and this elicits the evolution of free calcium carbonate after
a controlled flow rate under continuous but constant stirring. After calcination. The characteristic PO4 3− and OH− absorption bands
total addition of the reactants the suspension was transferred to a of HAP at 570, 600, 1033, 1085 and 3572 cm−1 , respectively were
stainless steel autoclave, sealed tightly and hydrothermally treated also observed for all samples. Dennis et al. [18] highlighted the fact
at 160 ◦ C for 10 h. The resultant precipitate was thoroughly washed that some cuticular vesicles of eggshell contain phosphate min-
with double distilled water, filtered and dried at 80 ◦ C for 2 h in an eral HAP (deposited by lipoprotein and protein–lipid complexes of
air oven. The dried precipitate was sintered at 400, 700 and 900 ◦ C, eggshell) and may be involved in the termination of calcite crys-
respectively for 2 h in an air furnace. tal growth. However, the intensities of these peaks are very less
since amount of HAP present in the crystal is very minimum (less
than 1%). The bending mode of water is positioned at 1650 and
2.2. Powder characterization 3440 cm−1 . It is very intense for raw eggshell and less intense for
the eggshell calcined at 800 and 900 ◦ C. It is also evident from the
FT-IR spectral analysis was performed for functional group anal- Fig. 1c that the 3 carbonate absorption band around 1400 cm−1
ysis of the samples by using Shimadzu model 8300. The FT-IR and 2 vibrational mode absorption at 875 cm−1 got disappeared
transmittance spectra of the samples were obtained in the range for the sample calcined at 900 ◦ C, which indicates the complete
400–4000 cm−1 using KBr pellet technique. All the samples were elimination of carbonate ion at this temperature.
identified by their X-ray diffraction pattern (XRD) using RICH- Fig. 2 shows the XRD pattern of eggshell calcined at 800 and
SEIFERT model 3000 X-ray diffractometer with Cu K␣ ( = 1.5406 Å) 900 ◦ C. The CaCO3 was completely decomposed and turned to CaO
incident radiation. The diffraction patterns were taken to the 2 at about 900 ◦ C. The phase analysis was performed using JCPDS card
ranges from 20◦ to 60◦ at a step sampling interval of 0.02◦ . Iden- no. 05-0586 for CaCO3 and 37–1497 for CaO. It is very obvious from
tification of phases were achieved by comparing the diffraction the figure that no other secondary phases were present except CaO
pattern of HAP with JCPDS standards of card no: 04-0932 for (Fig. 2) and CaCO3 (Fig. 2). Thus, the formation of CaO inferred from
HAP and 04-0733 for Ca(OH)2 . The size of the elementary crys- the corresponding XRD pattern elicits the loss of gaseous CO2 from
tallites was calculated using Scherrer’s formula. The full-width at CaCO3 at 900 ◦ C.
half-maximum of the 0 0 2 reflection was taken to measure the The formed calcium oxide when dispersed in double distilled
crystallite sizes of the HAP powder. water produces calcium hydroxide (confirmed by XRD shown in
The scanning electron microscopy (SEM) and energy disper- Fig. 2). The reaction can be represented as follows.
sive X-ray microanalysis (EDAX) analysis were performed for the
observation of microstructure and chemical constituent changes of CaO + H2 O → Ca(OH)2 (2)
the materials. The instrument Philips 501 scanning electron micro- The calcium hydroxide thus produced acts as one of the initial
scope equipped with EDAX was used for the microstructural and reactant (calcium source), which on reacting with orthophospho-
microanalysis. The samples were coated with a thin layer of gold ric acid to form HAP. The reaction theoretically expected for the
using an Edwards sputter coater S150B instrument. The EDAX spec- formation of HAP is
tra are acquired at an accelerating voltage of 100 kV. The elemental
composition (to provide Ca/P molar ratio of a selected area of the 10Ca(OH)2 + 6H3 PO4 → Ca10 (PO4 )6 (OH)2 + 18H2 O (3)
K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134 1129
Fig. 2. XRD pattern of (a) and (b) eggshell sintered at 800 and 900 ◦ C, respectively;
(c) Ca(OH)2 .
Fig. 4. FT-IR spectra of HAP synthesized using CTAB at the different temperatures for 10 h.
about same size. So it is reasonable to propose that when the reac- HAP crystals and hence CMC with CTAB:PO4 3− ratio = 1:1 was cho-
tion time is long enough, the final products would be invariably sen as the optimum CTAB concentration for the synthesis of highly
nano. Hence 10 h was chosen as the optimum ageing time for HAP crystalline HAP.
synthesis.
The effect of CTAB on HAP phases is shown in Fig. 5. The OH 3.2. Effect of sintering temperature
absorption peaks at 3570 and 630 cm−1 , OH stretching vibration
and OH structural vibration, decreased and disappeared with the Fig. 6 shows FT-IR absorption spectra of HAP powders heated
decrease in CTAB into the phosphate solution (Fig. 5a and b). The at 80, 400, 700 and 900 ◦ C. The infrared band positions and their
amount of CTAB remaining in the calcium phosphate samples assignments are summarized in Table 1. All the spectral sam-
was higher for CTAB at critical micelle concentration (CMC) with ples show vibrational modes corresponding to the phosphate and
CTAB:PO4 3− ratio = 1:1 as indicated by the intensity of absorption hydroxyl except carbonate groups. A broad envelope between
peaks at 2920 and 2851 cm−1 , and the intensity of OH absorp- 3200 and 3800 cm−1 due to the O–H stretch of water and HAP
tion bands got increased, which implied that CTAB concentrations was observed for the sample heated at 80 ◦ C. The O–H groups
below CMC could reduce the crystallization of HAP. The SEM pic- are hydrogen bonded. The peak at around 1650 cm−1 assigned to
tures of HAP synthesized at CMC and below CMC of CTAB (see bending mode of water was clearly visible for the sample heated
supplementary information) confirmed that the synthesis at CMC at 80 ◦ C. However, the intensities of these peaks were found to
improves crystallinity of the HAP powder. The results are in good decrease for those samples heated at higher temperatures. The
agreement with the results observed in FT-IR. Thus, it can be con- sharp peak at 3572 cm−1 observed for those samples heated at
cluded that the micelles act as nucleating points for the growth of higher temperatures is assigned to unhydrogen bonded free O–H
K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134 1131
Fig. 5. FT-IR spectra of HAP synthesized at 160 ◦ C for 10 h with the different
CTAB:PO4 3− ratio (a) 0.01:1, (b) 0.1:1 and (c) 1:1 (CTAB at CMC).
Table 2 of the particles during sintering. The particle sizes are ranging from
Elemental composition of nano-HAP powder.
100 to 300 nm and are largely agglomerated. The result is in good
Element Weight % Atom % agreement with the results obtained from SEM analysis. The reason
O 41.76 61.31 for the formation of nano-HAP with controlled microstructure can
Na 0.18 0.18 be explained as follows.
Mg 0.23 0.12 The addition of surfactant causes the controlled nucleation and
P 18.15 14.29 crystallization of the HAP during synthesis. The behavior of CTAB
Ca 39.18 23.85
was considered to correlate with the charge and stereochemistry
C 0.50 0.25
properties. In an aqueous system, CTAB would ionize completely
Total 100.00 100.00
and result in a cation with tetrahedral structure and forms micelles
at CMC. These micelles act as nucleating points for the growth of
HAP crystals with controlled structure. Meanwhile, the phosphate
where I3 0 0 is the intensity of (3 0 0) diffraction peak and V1 1 2/3 0 0 anion is also possessing tetrahedral structure. It has been reported
is the intensity of the hollow between (1 1 2) and (3 0 0) diffraction by several authors that a process called molecule recognition could
peaks of HAP. The evaluated degrees of crystallinity for the samples be realized at the inorganic/organic interface by the charge and
heated at 80 and 900 ◦ C was found to be 0.94 and 0.99. It is inferred stereochemistry complementarity. Hence it can be proposed that
from the resulted values that the increase in temperature improves the same charge and structure complementarity. The added surfac-
crystallinity of HAP. tant was then supposed to bind to certain faces of crystals and bind
The peak broadening of XRD reflection can be used to estimate to certain ions as well, so that ions could be incorporated to the
the crystallite size in a direction perpendicular to the crystallo- existing nuclei at a steady rate and the final shape and size could
graphic plane based on Sherrer’s formula as follows [21,22]: be well controlled. Further, the anisotropy of CTAB itself probably
0.9 induced the axis orientation growth of HAP.
Xs = (6) The high temperature and pressure of hydrothermal treatment
(ˇ cos )
could exert both effects on the final product. Firstly, they might
where Xs is the crystallite size (nm); the wavelength of monochro- cause some fibers to aggregate, but the interaction between the
matic X-ray beam (nm) ( = 0.15406 nm for Cu K␣ radiation); ˇ the aggregates is so weak that they tend to dissolve in the aqueous
full-width half-maximum for the diffraction peak under consid- system. Secondly, they might raise the solubility of HAP to some
eration (rad); and the diffraction angle (◦ ). The reflection plane extent and accelerate the dissolution and crystallization process.
(0 0 2) was chosen for calculation of the crystallite size since the Since the crystallization process is under critical control of CTAB,
peak appears at this Miller’s family plane is sharper and isolated the resulting HAP was invariably nano-sized.
from others and shows the crystal growth along the c-axis of the
HAP crystalline structure. The calculated crystallite size, Xs for HAP 4. Conclusions
powder heated at 80 ◦ C and 900 ◦ C at this diffraction peak using
Scherrer’s formula was found to be 34 and 88 nm, respectively. In the present work, nano-HAP with the particle size ranging
Thus, both the XRD and TG-DTA results have depicted that the sta- from about 100–300 nm was synthesized by hydrothermal method
bility of the HAP synthesized by hydrothermal method using CTAB from calcined eggshell using CTAB as regulator. Functional group
as a regulator was extended up to 1100 ◦ C and thereafter it was and phase identification corresponding to eggshell and HAP were
decomposed into -TCP. investigated by FT-IR and XRD studies. SEM and AFM techniques
SEM pictures of HAP powder heated at different tempera- (see supplementary information) employed to study the mor-
tures revealed that the synthesized powder appeared to be white phology of HAP have exhibited the formation of well-crystallized
crystals, bouncy and free flowing powder (see supplementary nano-particles with uniform shape. TG/DTA results have shown
information). The initial powders consist of small agglomerated that the formed nano-HAP was thermally stable up to 1100 ◦ C.
rod-like crystals, which upon sintering became deagglomerated The Ca/P ratio obtained from EDAX analysis confirmed the value of
and appeared as nano-rods. The morphology of HAP powder 1.67 corresponding to stoichiometric HAP. The observations further
sintered at 900 ◦ C indicates that most of the HAP particles are non- revealed that the crystallization process under critical micelle con-
agglomerated and monodispersive. It can be inferred that majority centration (CMC) of CTAB resulted in the production of nano-HAP
of the particles are of single crystals with regular shape and cleaner with uniform size. Moreover, the use of eggshell as a calcium source
contours. The phase constitution and chemical homogeneity of HAP represents a potential diminution in the costs of HAP production
were examined by EDAX analysis. Table 2 illustrates the elemen- with the additional advantage of recycling a waste product.
tal composition of the synthesized nano-HAP powder. The Ca/P
molar ratio was found to be 1.67, which indicates the formation Acknowledgement
of stoichiometric HAP. The result shows that the HAP is primarily
composed of Ca, P and O along with the trace amount of Na, Mg and The authors acknowledge the financial support rendered by
C. University Grants Commission (UGC) through University With
The AFM images (both 2D and 3D view) of the HAP synthe- Potential For Excellence (UWPFE) programme to carryout this
sized using CTAB by hydrothermal method have allowed making work.
a morphological interpretation of the crystal growth occurred after
sintering at 900 ◦ C (see supplementary information). The topogra- Appendix A. Supplementary data
phy of the HAP crystals is relatively smooth with a height variation
of ∼2.5 nm. The image shows the relevant variations in the crystal Supplementary data associated with this article can be found, in
growth recorded on 0.2 m × 0.2 m area of the sintered HAP pel- the online version, at doi:10.1016/j.saa.2009.09.021.
let. Both the 3D view and the 2D view depict the presence of several
plateaus with approximately same heights. Especially the 3D view
References
illustrates the maximum height of the plateau, which in turn elic-
its the height of the crystal growth. The crystals were observed to [1] H. Aoki, Science and Medical Applications of Hydroxyapaptite, Japanese Asso-
present well-defined grain boundaries due to non-agglomeration ciation of Apatite Science, Tokyo, Japan, 1991.
1134 K. Prabakaran, S. Rajeswari / Spectrochimica Acta Part A 74 (2009) 1127–1134
[2] W. Suchanek, M. Yoshimura, J. Mater. Res. 13 (1998) 94–117. [13] A.J. Ruys, M. Wel, C.C. Sorrell, M.R. Dickson, A. Brandwood, B.K. Milthorpe,
[3] J.B. Park, R.S. Lakes, Biomaterials—An Introduction, 2nd ed., Plenum Press, New Biomaterials 16 (1995) 409–415.
York, 1992. [14] M. Jarcho, C.H. Bolen, M.B. Thomas, J. Bobick, J.F. Kay, R.H. Doremus, J. Mater.
[4] J.D. de Bruijin, Y.P. Bovell, C.A. Van Blisterswijk, Biomaterials 15 (1994) Sci. 11 (1976) 2027–2035.
543–550. [15] M. Akao, H. Aoki, K. Kato, J. Mater. Sci. 16 (1981) 809–812.
[5] I.R. Gibson, S. Ke, S.M. Best, W. Bonfield, J. Mater. Sci. Mater. Med. 12 (2001) [16] T.S.B. Narasaraju, V.L.N. Rao, M. Lal, U.S. Rai, Indian J. Chem. 13 (1975) 369–378.
163–171. [17] A.J. Ruys, C.C. Sorrell, A. Brandwood, B.K. Milthorpe, J. Mater. Sci. Lett. 14 (1995)
[6] W.R. Rao, R.F. Boehm, J. Dent. Res. 53 (1974) 1351–1354. 744–747.
[7] P.E. Wang, T.K. Chaki, J. Mater. Sci. Mater. Med. 4 (1993) 150–158. [18] J.E. Dennis, S.Q. Xiao, M. Agarwal, D.J. Fink, A.H. Heuer, A.I. Caplan, J. Morphol.
[8] A. Royer, J.C. Viguie, M. Heughebaert, J.C. Heughebaert, J. Mater. Sci. Mater. Med. 228 (1996) 287–306.
4 (1993) 76–82. [19] J.A.M. van der Houwen, G. Cressey, B.A. Cressey, E. Valsami-Jones, J. Cryst.
[9] J.C. Knowles, W. Bonfield, J. Biomed. Mater. Res. 27 (1993) 1591–1598. Growth 249 (2003) 572–583.
[10] N. Patel, I.R. Gibson, S. Ke, S.M. Best, W. Bonfield, J. Mater. Sci. Mater. Med. 12 [20] E. Landi, A. Tampieri, G. Celotti, S. Spiro, J. Eur. Ceram. Soc. 20 (2000) 2377–
(2001) 181–188. 2387.
[11] M.G.S. Murray, J. Wang, C.B. Panton, P.M. Marquis, J. Mater. Sci. 30 (1995) [21] L. Slavica, Z. Slavica, M. Nada, M. Slobodan, Thermochim. Acta 374 (2001) 13–
3061–3074. 22.
[12] P. Van Landuyt, F. Li, J.P. Keustermans, J.M. Streydio, F. Delannay, E. Munting, J. [22] S. Zhang, K.E. Gonslaves, J. Mater. Sci. Mater. Med. 8 (1997) 25–28.
Mater. Sci. Mater. Med. 6 (1995) 8–13.