Synthesis of Biphasic Calcium Phosphate Made of Eggshells Through A Hydrothermal Method
SYNTHESIS OF BIPHASIC CALCIUM PHOSPHATE MADE
OF EGGSHELLS THROUGH A HYDROTHERMAL METHOD
NOLDY HUTAGALUNG
DEPARTMENT OF PHYSICS
FACULTY OF MATHEMATICS AND NATURAL SCIENCES
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
STATEMENT ON THE THESIS AND SOURCE OF
INFORMATION
I hereby declare that the thesis of “Synthesis of Biphasic Calcium
Phosphate Made of Eggshells Through A Hydrothermal Method” is my work with
the direction of supervising comitee and has not been submitted in any form to
any college. Source of information derived or quoted from the work or not
published by other authors mentioned in the text and listed in the refference at the
end of this thesis.
Bogor, September 2013
Noldy Hutagalung
Student ID G74090023
ABSTRACT
NOLDY HUTAGALUNG.Synthesis of Biphasic Calcium Phosphate Made of
Eggshells Through A Hydrothermal Method. Supervised by KIAGUS DAHLAN
and IRMANSYAH.
Biphasic Calcium Phosphate mixed of hydroxyapatite and tricalcium
phosphate. A hydrothermal method of synthesizing by mixing CaO as calcium
source and (NH4)2HPO4 as phosphate source with distilled water. CaO got from
calcination process made of eggshells. All samples were synthesized using
hydrothermal reactor then followed by sintering process. The molar ratio of Ca
and P for all samples was 0.5 M : 0.3 M. Hydrothermal temperature and sintering
temperature were variated they are 250 ºC, 300 ºC for hydrothermal and 800 ºC,
900 ºC, 1000 ºC for sintering, higher treatment temperature, tricalcium phosphate
appears. Final results of synthesis were characterized by using X-ray Diffraction
and Fourier Transform Infrared. X-ray Diffraction patterns was ecompared with
database Join Committee on Powder Diffraction Standards and found tricalcium
phosphate and hydoxyapatite.
Keywords : biphasic calcium phosphate, hydrothermal, hydroxyapatite, tricalcium
phosphate
NOLDY HUTAGALUNG.Sintesis Biphasic Calcium Phosphate Berbahan
Cangkang Telur dengan Metode Hidrotermal.Dibimbing oleh KIAGUS
DAHLAN dan IRMANSYAH.
Biphasic Calcium Phosphate terdiri dari hidroksiapatit dan trikalsium
posfat. Metode hidrotermal merupakan metode untuk mensintesis CaO sebagai
sumber kalsium dan (NH4)2HPO4 sebagai sumber posfat dengan aquades sebagai
pelarut. Semua sampel disintesis menggunakan reaktor hidrotermal kemudian
dilakukan proses sintering. Perbandingan molar antara Ca dan P untuk semua
sampel adalah 0.5 M : 0.3 M. Temperatur hidrotermal dan temperatur sintering
dilakukan variasi yakni 250 ºC, 300 ºC untuk temperatur hidrotermal dan 800 ºC,
900 ºC, 1000 ºC untuk proses sintering, semakin tinggi suhu maka fase trikalsium
posfat
semakin banyak jumlahnya. Hasil akhir dari sintesis kemudian
dikarakterisasi menggunakan X-ray Diffraction dan Fourier Transform Infrared.
Hasil karakterisasi menggunakan X-ray Diffraction dibandingkan dengan database
Join Committee on Powder Diffraction Standards kemudian diperoleh fase
trikalsium posfat dan hidroksiapatit.
Kata kunci : biphasic calcium phosphate, hidroksiapatit, hidrotermal, trikalcium
posfat
SYNTHESIS OF BIPHASIC CALCIUM PHOSPHATE MADE
OF EGGSHELLS THROUGH A HYDROTHERMAL METHOD
NOLDY HUTAGALUNG
Thesis
Submitted in Partial Fulfillment of the Requirements
for the Degree of Sarjana Sains
Department of Physics
DEPARTMENT OF PHYSICS
FACULTY OF MATHEMATICS AND NATURAL SCIENCES
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
Research Tittle : Synthesis of Biphasic Calcium Phosphate Made of Eggshells
Through A Hydrothermal Method
Name
: Noldy Hutagalung
Student ID
: G74090023
Approved by
Dr Kiagus Dahlan
Supervisor
Dr Irmansyah, MSi
Co Supervisor
Endorsed by
Dr Akhiruddin Maddu, MSi
Head of Physics Department
Date of graduate :
PREFACE
Praise to my Almighty God for His blessing I could finish this research
succesfully. Love and hug to my lovely family for support and pray me all the
time specially to my big mommy and daddy, to my sisters Helga, Maria, Yana,
and Eve, to my brohers Rommy, David, Elnoa for everything that you gave to
make me laugh. My respect and honour to my supervisors : Dr. Kiagus Dahlan,
Dr. Irmansyah, M.Si, and Setia Utami, M.Si for all moment to teach me how to do
this research well. I thank God to have friends like Jise Hari, Margaretha Septiana,
Ratna Sari, Saima Mega, Nasib Pols and PARTARU members for support me all
the time. My big thank to 46 Physics student and biomaterialler : Upri, Indri,
Budi, Kak Aisyah, Mita for helping me while doing this research, then to Pena
stationary : Dede, Vina, Rady and Kania, my dorm mates : Ade, Muga, and Desy.
The author hopes much suggestions to make this research will be better and author
do hope this research will be usefull to everyone.
Bogor, September 2013
Noldy Hutagalung
CONTENTS
LIST OF TABLES
vi
LIST OF FIGURES
vi
LIST OF APPENDICES
vi
INTRODUCTION
1
Background
3
Objectives of Research
3
Purposes of Research
3
Usefulness of Research
3
Scope of Research
3
Hypothesis
3
EXPRIMENTAL METHOD
3
Material and Equipment
3
Preparation of Eggshell
4
Preparation of Sample
4
Hydrothermal Method
5
Sintering Process
6
Sample Characterization by x-ray diffraction (XRD)
7
Sample Characterization by fourier transform infrared (FTIR)
7
RESULT AND DISCUSSION
8
Calcination Process and synthesis of BCP
8
Hydrothermal and Sintering Process
8
Phase, average crystallite size (ACS), and lattice parameter of BCP 9
Phase Groups of BCP
12
Phase of HA with and without Sintering Process
13
CONCLUSION AND SUGGESTIONS
15
Conclution
15
Suggestions
15
REFERENCES
16
APPENDICES
18
CURICULUM VITAE
29
LIST OF TABELS
1 Variation of hydrothermal temperature, sintering temperature and
sintering time
2 Mass of CaO after sintering process
3 Mass of all samples after sintering
4 Ratio of HA and -TCP
5 Parameter lattice of HA
6 Atomic crystall size of all sample
6
8
9
11
12
12
LIST OF FIGURES
1 Preparation then followed by calcination process
2 Solution and suspension preparation followed by stirring and dropping
process
3 Hydrothermal process followed by aging
4 Filtration followed by sintering process
5 XRD patterns of A1-A3 samples with variations of treatment
6 XRD patterns of A4-A6 samples with variations of treatment
7 FTIR spectra of A1-A3 samples with variations of treatment
8 FTIR spectra of A4-A6 samples with variations of treatment
9 XRD patterns of HA1
10 XRD patterns of HA2
4
5
6
7
10
11
13
13
14
15
LIST OF APPENDICES
11 Flowchart
12 Data JCPDS (Join Committee on Powder Diffraction Standards)
13 XRD patterns and FTIR spectra
19
20
22
INTRODUCION
Background
The bone damage is a serious health problem because bone is one of the
most important organs of human. However, bone damage is not only caused by
work accidents but diseases can cause bone damage. Some deseases caused bone
damage are osteoporoses, bone tumors, certain cancer or a brittle bone desease
called osteogenesis.1,2 The bone implantation is the way to treat bone damage and
while doing this we need appropriate materials. The composition of material that
we will use has to similar with bone composition.
In this era, the development of biomaterial not only talked about synthesis
but and characterization but new challanges also. Researchers not only interest
focuses primarily on synthesis and characterization, biological behavior of
material and porouess are important to know. In order to develop material must
have support knowledge of different disciplines and made a colaboration.3 In the
following some interdisiplinaries research that support are biochemistry,
biomedical engineering and medical sciences.
An ideal biomaterials that used for implantation should be has biological
behaviors such as bioactive, biodegradable, non-toxic and biocompatible with
human body.1,2
A biomaterial is a synthetic material used to replace part of living system or
to function in intimate contact with living tissue.3 Biomaterial has been
acknowledged as a bone graft material in a range of medical and dental
applications due to their similar chemical composition with natural bones.
Generally, bone substitution materials such as autograft, allograft and xenograft
are used to solve problems related to bone trauma and fractures. Material which
obtained from the human involved called autograft, from the other human called
allograft and from animal called xenograft.2 But, none of these materials provide a
perfect replacement of the bone due to mechanical and biological instability and
incompatibility. Recently, calcium phosphate bioceramics such as calcium
phosphate, tri-calcium phosphate and hydroxyapatite are identified as most
suitable bone substitution materials to serve the demand. Unlike other calcium
phosphates, HA does not break under physiological conditions. In fact, it is
thermodynamically stable at physiological pH and actively takes part in bone
bonding.4
A biomaterial is an inert material which implanted into living systems to
change living tissue or organ function. The purposes of biomaterials are used to
implant in living sytems’s bone are repair, replace and restore organ or tissue.
Biomaterials that developed widely are hydroxyapatite, -TCP and material
mixtures of HA and -TCP is called biphasic calcium phosphate.
2
phosphate, hydroxyapatite and biphasic calcium phosphate has difference
biological behavior each other.3
Hydroxyapatite is the most commonly used calcium phosphate in the
medical fields or dentistry, as it posseses excellent biocompatibility and
osteoconductive. It has chemical formula Ca10(PO4)6(OH)2and a Ca/P ratio of 1.67.
The density of this material in theoritical is 3.156 g/cm3. The crystalinity of this
material has hexagonal rhombic prisms with the space group being P63/m. Its
units cell contains a complete representation of the apatite crystal, consisting of
Ca2+, PO43-, and OH- groups.2,3 The biological behaviors of this material are bioinert and resorbable. Synthesis of hydroxyapatite can be prepared by using a
variety of method and precursor. The variation of method are wet method and dry
method. Wet method by precipitation by involving the neutral reaction of acid and
alkaline solutions or calcium salts and phosphate salts, dry method by heating,
and hydrothermal method.1
Tricalcium phosphate was used as material to synthesize BCP with HA.
The chemical formula of tricalcium phosphate is Ca3(PO4)2. TCP has four
polymorphs are α, , , and super-α. –TCP is stable up to 1125 °C but above
11β5 °C up to 14γ0 °C α becomes the stable phase. Super-α foms between
14γ0°C and the melting point at 1756 °C. –TCP has the rhombohedral space
group R3c with unit cell a=1.0439 nm, c=3.7375 nm (hexagonal setting) with 21
formula units per hexagonal unit cell. The dissolution rate of –TCP was three
times higher than that of HA. The dissolution rate increased shown like this : HA
< –TCP < α-TCP < TTCP.1,3,5 The biological behaviors of this material are
bioactive, bioresorbable, and biodegradable.
HA and TCP have been used, not only for bone implant substitute materials,
but have also been identified for applicability as drugs delivery, antiseptics, food
supplements, and hypothermia treatment agents. The potential of the material as
anticancer agents has been investigated and it has been found that nanoscale HA is
effective in inhibiting cancer cells growth.6 HA and TCP although they have
similar chemical composition but they different in their biological resorbing
capacity. The mixture material of HA and –TCP is called bihasic calcium
phosphate (BCP). Development of BCP especially with HA and TCP has drawn
considerable attention.7-10 In medical application they have found as bone graft
substitute. The resorbability of BCP ceramics depends on ratio of HA and –TCP,
hight ratio will produce high resorbability.
Some kinds of phosphate have been used to react with CaO of some sources
such as eggshell, fishbone, seashell, etc. Kinds of phospate which generally used
are H3PO4, (NH4)2HPO4, and Na2HPO4.1-12 The eggshell represents the 11% of
the total weight of the egg and is composed by calcium carbonate (94%), calcium
phosphate (1%), orghanic matter (4%) and magnesium carbonate(1%).12,13
3
Objectives of Research
1
2
What the effects of variation of hydrothermal-temperature, hydrothermalholding time and sintering-temperature to produce biphasic calcium
phosphate with ratio of hyrdoxyapatite and -tricalcium phosphate ?
Hows the final result of characterization by using X-ray diffraction (XRD)
and fourier transform infrared (FTIR) ?
Purposes of Research
1
2
3
To synthesize biphasic calcium phosphate using eggshells
To characterize biphasic calcium phosphate by using XRD and FTIR
To get BCP with ratio of HA and -TCP are 60 : 40 and 70 : 30
Usefulness of Research
The useful of this research was to developed biomaterial as bone graft and
mixed two different phase which is called biphasic and obtained new calcium
source from eggshells.
Scope of Research
The aim of this research was to synthesize biphasic calcium phosphate by
hydrothermal method. Calcium source got from eggshells by calcination process
by freeing CO2 and (NH4)2HPO4 as phosphate source. Sample was characterized
by X-ray Diffraction and Fourier Transform Infrared.
Hypothesis
Variation of hydrothermal-temperature, hydrothermal holding time and
sintering temperature can produce BCP with ratio of HA and -TCP are 60 : 40
and 70 : 30. The ratio as mentioned above can produce an ideal material to
implant to human or other living system.
EXPERIMENTAL METHOD
Material and Equipment
The materials which used in this research made of eggshells in chemical
form CaCO3 was calcinated by freeing CO2 with final result of calcination was
CaO as calcium source, (NH4)2HPO4 as phosphate source, aquades (H2O). These
are the equipment : beaker glass, stirrer equipment, furnace, precipitation.
4
equipment, and hydrothermal reactor. Samples was characterized by using X-ray
Difraction and Fourier Transform Infrared.
Preparation of Eggshell
Washing eggshell to eliminate inner membrane of eggshells by using
aquades, then use furnace to calcinate it in 1000°C at heating rate 5°C/minute for
5 hours. At this process eggshel transform from calcium carbonate into calcium
monoxide by freeing carbon dioxide (CO2).14 The material which produce of
calcination process is CaO (calcium monoxide). Sample preparation and
calcination process are shown by the Figure 1 and the chemical reaction of this
process is shown below.
CaCO3 → CaO + CO2
Eggshells putted
into crucible
Calcination T=1000 °C
for 5 hours
CaO after
calcination process
CaO after grinding
Figure 1 Preparation then followed by calcination process
Preparation of Sample
This reasearch starting from suspens of calcium preparation from 0.5 M
CaO was mixed with 100 ml water destilled. Then phosphate solution preparation
from 0.3 M (NH4)2HPO4 was mixed with 100 ml water destilled. Suspens of
5
calcium was stirred by using stirrer equipment at 300 rpm for 20 minutes. At 5
minutes left after stirrer was ran then mix calcium solution and phosphate solution
by using dropping equipment until stirrer stop. Calcium solution, phosphate
suspensions followed by stirring and dropping process are shown by the Figure 2
and the chemical reaction of mixing process is shown bellow :
10CaO + 14H2O + 6(NH4)2HPO4 → Ca10(PO4)6(OH)2 + 18H2O
0.3 M (NH4)2HPO4
+ H2O
0.5 M CaO + H2O
Stirring and
dropping procces
Mixed of calcium suspension
and phosphate solution
Figure 2 Solution and suspension preparation followed by stirring and dropping
process
Hydrothermal Method
The mixed of solution and suspension moved in hydrothermal reactor.
Hydrothermal method is one of methods which considered by researchers to
synthesize calcium monoxide be a biomaterial which has high quality, high
crystalinity and high reactivity. Hydrothermal temperature and sintering
temperature was variated while synthesizing sample. The variation of treatment is
shown in Table 1.
6
Tabel 1 Variation of hydrothermal temperature, sintering temperature, and
sintering time
Sample
code
A1
A2
A3
A4
A5
A6
HA1
HA2
Hydrothermal
temperature
(°C)
250
250
250
300
300
300
150
200
Hydrothermal
rpm (rpm)
300
300
300
300
300
300
300
300
Sintering
temperature
(°C)
800
900
1000
800
900
1000
800
-
Sintering
time
(hours)
5
5
5
5
5
5
5
-
Hydrothermal
pressure (psi)
245
245
245
510
510
510
242
246
The holding time for all samples were 10 hours but special for HA2 was not
sintered and the holding time was 24 hours followed by drying process in 50 ºC
for 8 hours. After all process done then solutions was kept in hydrothermal tube
for overnight or generally this process is called “aging”. Hydrothermal process
and aging are shown by the Figure 3.
Suspension
Hydrothermal
process + aging
Figure 3 Hydrothermal process followed by aging
Sintering Process
Sintering process starting from dropping process. Aging process produced
final solution then solution was filtrated. The residue of solution then moved into
crucible then put in to furnace at 800°C, 900°C, 1000°C at heating rate
5 °C/minute for 5 hours but sample must be dried first at 110 °C with heating rate
5 °C/minute for 5 hours. Filtration and sintering process are shown by the Figure
4.
7
Filtration
Sintering process
Sample after
sintering process
Sample after grinding
process
Figure 4 Filtration followed by sintering process
Sample Characterization by X-ray Diffraction
All samples characterized by using X-ray diffraction GBC EMMA. 2 gr of
each sample putted on holder then setted sample’s name, initial angel 10º, final
angle 80º, CuKα = 1.54056 and analysis speed of computer. The purpose of this
process was to identify crystal structure and lattice parameter of each sample.
Sample Characterization by Fourier Transfrorm Infrared
All samples charcterized by using Fourier Transform Infrared ABB MB
3000. The FTIR spectra used KBr pellet technique. 200 mg KBr mixed with 2 mg
of each sample. The fourier transform infrared spectroscopy employed to
characterize the different functional groups of hydroxyapatite and tricalcium
phosphate. The FTIR used vibration energy of functional groups of HA and TCP
such as PO43-groups, CO32- groups and OH- groups.
8
RESULTS AND DISCUSSION
Calcination Process and synthesis of BCP
The variety of method to synthesize powder are wet and dry method. Wet
method by precipitation by involving the neutral reaction of acid and alkaline
solutions or calcium salts and phosphate salts, dry method by heating, and
hydrothermal method.1 Two kinds of raw material which commonly used to
synthesize BCP are synthetic material and natural material. In this research as
calcium source was waste material it was eggshells.
Calcination was the process to get CaO from CaCO3 by using furnace in
1000 °C at heating rate 5°C/minute for 5 hours called optimum condition of
caltination. At this process eggshells transform from calcium carbonate into
calcium monoxide by freeing carbon dioxide (CO2). Chemical reaction of
calcination process is shown below :
CaCO3 → CaO + CO2
CO2 can be eliminated at higher temperature than 750 ºC, Magnesium (Mg)
can be eliminated in 650 ºC, and organic composition can be eliminated in 80 ºC –
100 ºC.1
The synthesis of BCP produced calcium phosphate ceramics which are
have an important position among other biomaterials, because they are considered
to be almost fully biocompatible with living bodies when replacing the hard bone
tissues, they are hydroxyapatite (Ca10(PO4)6(OH)2) and tricalcium phosphate
(Ca3(PO4)2 ) are currently recognized as ceramic materials that significantly
simulate the mineralogical structure of bone. In this research the molarity ratio of
Ca and P was 0.5 M : 0.3 M because concentration ratio of calcium and phosphate
of HA was 1.67. The reaction of synthesis in hydrothermal reactor as shown
below:
10CaO + 14H2O + 6(NH4)2HPO4 → Ca10(PO4)6(OH)2 + 18H2O
Product of calcination process was CaO and the mass of CaO is shown in Table 2.
Table 2 Mass of CaO after calcination process
Mass of Ca(CO)3 (gr)
21.896
22.722
25.172
22.963
21.666
21.395
20.266
24.716
27.059
27.185
18.954
19.963
Mass of CaO (gr)
11.524
12.083
13.592
12.453
11.453
11.693
10.893
13.294
14.546
14.500
9.914
10.651
9
Hydrothermal and Sintering Process
As mentioned before two kinds of powder synthesis are wet method by
precipitation by involving the neutral reaction of acid and alkaline solutions or
calcium salts and phosphate salts, dry method by heating, and hydrothermal
method. HA prepared by the wet method in atmosphere consist of very small
crystal with lattice deffects. The way to obtain large, perfect, single crystal of HA,
it is preferable to use the hydrothermal technique.3
Hydrothermal synthesis is characterized by the reaction of aqueous
solution in closed recipients under controlled temperature and/or pressure. The
temperature can be elevated above the boiling point of water, reaching the
pressure of vapor saturation. Hydrothermal method is the best method to get
biomaterial high quality, homogenous, high purity, high crystalinity, and high
reactivity. There are three main variables in hydrothermal process they are
temperature, pressure, and chemist potential. The lattice parameter of HA
prepared by the hydrothermal method depend on the reaction temperature and
pressure.1
Sintering was the process in high temperature to appear phase of material
but not all powder synthesises has to passed this process. The use of natural
material such as seashells, eggshells and coral for synthesizing are commonly has
to passed sintering process. Although sample had passed hydrothermal process
with variety of temperature but it had no effect to appeared phase of material
because hydrothermal process just for creating ion groups of HA and -TCP.
Sintering process starting from dropping process. Aging process produced
final solution then solution was filtrated. The residue of solution then moved into
crucible then put in to furnace at 800 °C, 900 °C, 1000 °C at heating rate
5 °C/minute for 5 hours but sample must be dried first to freeing H2O of
suspensions at 110 °C with heating rate 5 °C/minute for 5 hours.
Based on chemical reaction, the result of synthesis was HA but by
sintering with high temperature samples might contain TCP because with
increasing temperature, the apatite structure changes to a new space group and
TCP appears and if in sample contain two different phase it is called biphasic and
in this case two different phase were HA and -TCP which is called BCP. The
mass of all samples are shown in Table 3.
Table 3 Mass of all samples after sintering process
Sample Code
A1
A2
A3
A4
A5
A6
HA1
HA2
Mass (gr)
3.4005
3.3089
3.3330
3.9313
3.7102
3.7103
3.8345
3.3043
10
Phase, Lattice Parameter, and Average Crystallite Size of BCP
The result of XRD characterization for all samples were compared with
the data JCPD (Join Committee on Powder Diffraction Standards). The result of
compared for sample code A1- A6 are shown by the Figure 5 and 6.
Figure 5 XRD patterns of A1 – A3 samples with variations of treatment
Figure 6 XRD patterns of A4 – A6 samples with variations of treatment
11
All figures above show that the result of XRD were same generally. Figure
5 shows the result of sample A1, A2, A3, they were same in hydrothermal
temperature β50 ºC but they are different in sintering temperature. That’s why
their characteristic were different each other. According to thermal stability of
HA and –TCP, with increasing temperature, the apatite structure changes to a
new space group and TCP apprears.3 As mentioned before –TCP is stable up to
11β5°C which is means higher temperature, –TCP appears . Figure 6 shows the
result of sample A4, A5, A6, they were same in hydrothermal temperature at 300
ºC but they were different in sintering temperature.
Calcium phosphate bio ceramics such as calcium phosphate, tricalcium
phosphate and hydroxyapatite has been acknowledged as a bone graft material in
a range of medical and dental applications due to their similar chemical
composition with natural bones. Generally, bone substitution materials such as
autograft, allograft and xenograft are used to solve problems related to bone
trauma and fractures. They are identified as most suitable bone substitution
materials to serve the demand. According to result of evaluated based on data
JCPDS researcher got nothing sample which contained pure HA and –TCP ,
researcher got apatite calcium type A (CAA) and octacal cium phosphate (OCP)
but majority pahses in each sample were HA and –TCP. The existence of OCP
did not matter for ceramics used for implantation because synthetic OCP showed
its osteoconductive characteristics in the bone marrow space such as enhance bone
regeneration at the initial bone apposition stage and stimulate resorption of the
newly formed bone. In addition, biodegrade rate of synthetic OCP is faster than TCP ceramics and OCP enchance bone formation more than HA and -TCP
ceramics when implanted in living system.13
XRD patterns were used to find phase of material, parameter lattice and
atomic crystal size. XRD patterns were compared with data JCDPS and based on
result of XRD patterns above we knew that they are different each other specially
for ratio of HA and –TCP. The strongest peak of all samples is appears at 31.7º
and it is belong to HA. The result of calculated are shown in Table 4.
Table 4 Ratio of HA and –TCP
Sample Code
A1
A2
A3
A4
A5
A6
HA : –TCP
64: 36
66 : 34
62 : 38
74 : 26
63 : 37
63 : 37
The result of comparing XRD patterns and data JCPDS were used to
calculated parameter lattice. The crystallite size of all samples based on full width
half maximum (FWHM) of X-ray diffraction patterns were calculated by using
12
MDI Jade 6 program. The lattice parameter and average crystallite size are shown
in Table 5 and 6.
Table 5 Lattice parameter of HA
Sample code
A1
A2
A3
A4
A5
A6
HA1
HA2
a (Å)
10.056
9.519
9.741
9.427
11.738
9.407
9.414
9.468
Table 6 Average crystallite size of all samples
Sample code
A1
A2
A3
A4
A5
A6
HA1
HA2
c (Å)
7.352
6.959
7.127
6.893
8.510
6.881
6.878
6.925
ACS (nm)
42.890
49.562
54.298
49.180
40.340
53.810
49.332
34.911
The lattice constant of HA prepared by the hydrothermal method depend
on the reaction temperature and pressure.1 According to the data that we got while
synthesizing BCP higher temperature hydrothermal, higher pressure. The lattice
parameters of the synthesized BCP samples were obtained from the diffraction
pattern by fitting the peaks of identified reflections. Generally crystallite size of
all samples were similar except HA2 because optimum phase of hydroxyapatite
would be appeared in high temperature because CaO that we used in this synthesis
was wastes material.
Phase Groups of BCP
Previous report clearly demonstrated that infrared spectroscopy is an
indispensable tool in order to learn some special features of the preparation of
calcium hydroxyapatite ceramics.10 Therefore, Fourier Transform Infra red
characterization was carried out for BCP samples to study the spectral
characteristics which indicated the chemical bonding. The result of FTIR
characterization for all sample are shown by the Figure 7 and 8.
13
Figure 7 FTIR spectra of A1 – A3 samples with variation of treatment
Figure 8 FTIR spectra of A4 – A6 samples with variation of treatment
14
The spectra indicates the formation of a typical HA structure containing
PO43- band and OH- band, TCP structure containing PO43- band, and OCP
structure containing HPO42- band. Phosphate characteristic absorption bands
appear at wavenumber 650 - 550 cm-1 is called v4, 1200 – 900 cm-1 is called v3,
430 – 460 cm-1 is called v2 and around 960 cm-1 is called v1. Carbonate absorption
bands between 1600 – 1400 cm-1 is called v1 and around 870 cm-1 is called v2.
Absorption bands around 632 cm-1, 3573 cm-1, and 3649 cm-1 show the OH- group.
Absorption band around 651 cm-1 and 3445 cm-1 show H2O that absorbed by
sample.1,15,16
Phase of HA with and without Sintering Process
In this research two different samples were addded to synthesize as shown
in Table 1 which are different in treatment of variation. For HA1 was
hydroxyapatite with temperature of hydrothermal is 150 ºC, hydrothermal holding
time is 3 hours, and sintered in 800 ºC for 5 hours. Another one was
hydroxyapatite with temperature of hydrothermal 200 ºC, hydrothermal holding
time 24 hours without sintering process (HA2). XRD patterns were compared
with the data JCPDS. The result of these synthesises were different each other
specially for peak strong. HA1 had a stronger peak of all peak in this sample at
31.7° with intensity at 385 counts/s. HA2 had stronger peak at 31.7º with intensity
at 267 counts/s. The result of compared for two samples hydroxyapatite are
shown by the Figure 9 and 10.
Figure 9 XRD patterns of HA1
15
Figure 10 XRD patterns of HA2
Figure 10 shows the sample which totally different from those sample
which sinterred before. The main reason is because HA phase by using wastes
material such as eggshells, seashells and others wastes material will be created if
we sintered in high temperature. When hydrothermal process was run sample
synthesized in 250 ºC for 24 hours and analyzed result by using database JCPDS
we got OCP, HA, and -TCP and majority phase is HA. But if we compared result
analys of HA2 and sample with sintering process, sample with sintering better
than HA2 because the crysstalline structure of this sample was amorphous as we
see in Table 6. Otherwise if we use calcium nitrate tetrahydrate (Ca(NO3)2.4H2O)
or other synthetic materials the synthesized did not need sintering process.10
16
CONCLUSION AND SUGGESTIONS
Conclusion
This research was to synthesize BCP made of eggshells through a
hydrothermal method. Biphasic calcium phosphate was mixed of HA and -TCP.
Calcium source was from waste materials it was hen’s eggshells. The chemical
form of eggshells is CaCO3. Calcination was the process to get CaO by freeing
CO2 from CaCO3 using furnace at 1000 °C with heating rate 5 °C/minute for 5
hours. Each sample was mixed of calcium suspension and phosphate solution.
Ratio molar of Ca and P was 0.5 M : 0.3 M to synthesize BCP. Hydrothermal was
the method to synthesize calcium monoxide be a biomaterial high quality, high
crystalinity and high reactivity. Hydrothermal temperature and sintering
temperature were variated. For sintering process higher temperature, tricalcium
phosphate appears and for hydrothermal process higher treatment of temperature,
higher pressure in hydrothermal tube. Three main variables in hydrothermal
process were temperature, pressure, and chemist potential.
Product of sintering process was characterized by using XRD and FTIR.
XRD patterns were compared with the data JCPDS to know what the phase of
product. The result of comparing process was majority phases were HA and TCP. The lattice parameter and average cryztallite size were calculated by
comparing XRD pattern with database JCPDS. Higher treatment sintering
temperature had effect on atomic crystal size of BCP. To support the calculated
result of characterized by using XRD we got functional groups of HA and TCPas PO43-groups, CO32- groups and OH- groups of FTIR characterized.
Suggestions
1. As mentioned before with increasing temperature, the apatite structure
changes to a new space group and TCP apprears that’s why we suggest touse
sintering temperature higher than 1000 ºC to get more various of ratio of HA
and -TCP.
2. In this research all samples were characterized by using XRD and FTIR, the
author recomend to use SEM to know about the morphologh structure of BCP.
3. To get sample without impurities the residue of filtration must be washed by
using water destilled.
REFERENCES
1.
2.
Fajriah HA.Hydrothermal Synthesis of Biphasic Calcium Phosphate.[Thesis].
Bogor. Faculty of Mathematics and Natural Sciences, Bogor agricultural
University.2008;1-15.
Utami SD.Pembuatan Komposit Kalsium Fosfat-Kitosan dengan Metode
Sonikasi.[Thesis].Bogor. Fakultas Matematika dan Ilmu Pengetahuan
Alam,Institut Pertanian Bogor.2009;1,4-8.
17
3.
4.
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8.
9.
10.
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14.
15.
16.
Shi D.Biomaterials and Tissue Engineering.New York:Springer.2003;4-15
Sudip M, Biswanath M, Apurba D, Sudit SM.Studies on Processing and
Characterization of Hydroxyapatite Biomaterialsfrom Different Bio Wastes.
Journal
of
Minerals
&
Materials
Characterization
&
Engineering.2012;11(1):55-67.
Mohd AAMN, Maryam MR, Zainal AA.Synthesis and Characterization of Tricalcium Phosphate Ceramics Via Sol-Gel Method.Journal of Nuclear And
Related Technologies.2009;6(1):199-205.
Asep SFA, Iis S.Low temperature hydrothermal synthesis of calcium
phophate ceramics: Effect of excess Ca precursor on phase behaviour.Indian
Journal of Chemistry.2009;48A:1492-1500.
Victoria EC, Gnanam FD.Synthesis and Characterisation of Biphasic Calcium
Phosphate.Trends Biomater.Artif.Organs.2002;16(1):12-14.
Nezahat K, Cuneyt TA.Synthesis of Calcium Hydroxyapatite-Tricalcium
Phophate (HA-TCP) Composite Bioceramic Powders and Their Sintering
Behaviour.J.Am.Cetam.Soc.1998;81(9):2245-2252.
Zyman ZZ, Tkachenko MV, Polevodin DV.Preparation and Characterization
of Bihasic Calcium Phosphate Ceramics of Desired Composition.J Mater Sci:
Mater Med.2008;19:2819-2825.
Nuzulia NA.Study of Biphasic Calcium Phosphate Ceramics and HAChitosan Composite Implanted Into Sheep’s bone.[Thesis].Bogor.Faculty of
Mathematics and Natural Sciences, Bogor agricultural University;2009.
Earl JS, Wood DJ, Milne SJ.Hydothermal Synthesis of Hydroxyapatite.
Journal of Physics: Confrence series.2006;26:268-271.
El BH, Ginebra MP, Vert M, Boudeville P.Wet or Dry Mechanochemical
Syhthesis of Calcium Phosphates? Influence of The Water content on DCPDCaO Reaction Kinetics.Acta Biomaterialia.2008;4:378-386.
Sasikumar S, Vijayaraghavan R.Low Temperature Synthesis of
Nanocrystaline Hydoxyapatite from Egg Shells by Combustion
Method.Trends Bioamater. Artif. Organs.2006;19(2):70-73.
Eric MR, Miguel A, Witold B, Victor MC, Diaz EJR, Hernandez R, Rogelio
RJ.Synthesis of Hydroxyapatite fro Eggshell.Material Letters.1999;41.128134.
Liga BC, Natalija B.Research of Calcium Phosphates Using Fourier
Transform Infra Red Spectroscopy.[www.intechnopen]. Riga Technical
University.
Rozia AR, Rohana A, Mohammad AB, Sam’an MM.Synthesis and
Characterisation of pure Nanoporous Hydroxyapatite.Journal of Physical
Science.2011;22(1):25-37.
18
APPENDICES
19
Appendix 1 Flowchart
Materials and equipment preparation
Ready
No
Sample preparation
Hydrothermal synthesis
CaO, (NH4)2HPO4, H2O
Sintering
Biphasic calcium phospate
FTIR and XRD characterization
Data analysis
Report arrangement
Figure A7 Steps which author did as basic principle to research
20
Appendix 2 Data JCPDS of (a) -TCP, (b) HA, (c) OCP and (d) CAA
(a)
(b)
21
(c)
(d)
22
Appendix 3 XRD pattern of sample A1
XRD pattern of sample A2
23
XRD pattern of sample A3
XRD pattern of sample A4
24
XRD pattern of sample A5
XRD pattern of sample A6
25
XRD pattern of sample HA1
XRD pattern of sample HA2
26
FTIR spectra of sample
FTIR spectra of sample A2
27
FTIR spectra of sample A3
FTIR spectra of sample A4
FTIR spectra of sample A4
28
FTIR spectra of sample A5
FTIR spectra of sample A6
29
CURICULUM VITAE
The author was born in Sipoholon on December 23rd 1991 as fifth child of
eight from merried of Jodi Hutagalung and Marlince Sinaga. The writer was
educated at Elementary school SDN 0178492 in 1997-2003, Junior High school at
SMPN 1 Sipoholon in 2003-2006, Senior High School at SMAN 1 Sipoholon in
2006-2009. The author was accepted at Bogor Agricultural University in 2009 by
means of USMI (Ujian Seleksi Masuk IPB) in departmentof Physics . First year
the writer passed Tingkat Persiapan Bersama (TPB) then second year the writer
studied as student in department of Physics. The writer was one of laboratory
assistant of 1st Physics experiment.
OF EGGSHELLS THROUGH A HYDROTHERMAL METHOD
NOLDY HUTAGALUNG
DEPARTMENT OF PHYSICS
FACULTY OF MATHEMATICS AND NATURAL SCIENCES
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
STATEMENT ON THE THESIS AND SOURCE OF
INFORMATION
I hereby declare that the thesis of “Synthesis of Biphasic Calcium
Phosphate Made of Eggshells Through A Hydrothermal Method” is my work with
the direction of supervising comitee and has not been submitted in any form to
any college. Source of information derived or quoted from the work or not
published by other authors mentioned in the text and listed in the refference at the
end of this thesis.
Bogor, September 2013
Noldy Hutagalung
Student ID G74090023
ABSTRACT
NOLDY HUTAGALUNG.Synthesis of Biphasic Calcium Phosphate Made of
Eggshells Through A Hydrothermal Method. Supervised by KIAGUS DAHLAN
and IRMANSYAH.
Biphasic Calcium Phosphate mixed of hydroxyapatite and tricalcium
phosphate. A hydrothermal method of synthesizing by mixing CaO as calcium
source and (NH4)2HPO4 as phosphate source with distilled water. CaO got from
calcination process made of eggshells. All samples were synthesized using
hydrothermal reactor then followed by sintering process. The molar ratio of Ca
and P for all samples was 0.5 M : 0.3 M. Hydrothermal temperature and sintering
temperature were variated they are 250 ºC, 300 ºC for hydrothermal and 800 ºC,
900 ºC, 1000 ºC for sintering, higher treatment temperature, tricalcium phosphate
appears. Final results of synthesis were characterized by using X-ray Diffraction
and Fourier Transform Infrared. X-ray Diffraction patterns was ecompared with
database Join Committee on Powder Diffraction Standards and found tricalcium
phosphate and hydoxyapatite.
Keywords : biphasic calcium phosphate, hydrothermal, hydroxyapatite, tricalcium
phosphate
NOLDY HUTAGALUNG.Sintesis Biphasic Calcium Phosphate Berbahan
Cangkang Telur dengan Metode Hidrotermal.Dibimbing oleh KIAGUS
DAHLAN dan IRMANSYAH.
Biphasic Calcium Phosphate terdiri dari hidroksiapatit dan trikalsium
posfat. Metode hidrotermal merupakan metode untuk mensintesis CaO sebagai
sumber kalsium dan (NH4)2HPO4 sebagai sumber posfat dengan aquades sebagai
pelarut. Semua sampel disintesis menggunakan reaktor hidrotermal kemudian
dilakukan proses sintering. Perbandingan molar antara Ca dan P untuk semua
sampel adalah 0.5 M : 0.3 M. Temperatur hidrotermal dan temperatur sintering
dilakukan variasi yakni 250 ºC, 300 ºC untuk temperatur hidrotermal dan 800 ºC,
900 ºC, 1000 ºC untuk proses sintering, semakin tinggi suhu maka fase trikalsium
posfat
semakin banyak jumlahnya. Hasil akhir dari sintesis kemudian
dikarakterisasi menggunakan X-ray Diffraction dan Fourier Transform Infrared.
Hasil karakterisasi menggunakan X-ray Diffraction dibandingkan dengan database
Join Committee on Powder Diffraction Standards kemudian diperoleh fase
trikalsium posfat dan hidroksiapatit.
Kata kunci : biphasic calcium phosphate, hidroksiapatit, hidrotermal, trikalcium
posfat
SYNTHESIS OF BIPHASIC CALCIUM PHOSPHATE MADE
OF EGGSHELLS THROUGH A HYDROTHERMAL METHOD
NOLDY HUTAGALUNG
Thesis
Submitted in Partial Fulfillment of the Requirements
for the Degree of Sarjana Sains
Department of Physics
DEPARTMENT OF PHYSICS
FACULTY OF MATHEMATICS AND NATURAL SCIENCES
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
Research Tittle : Synthesis of Biphasic Calcium Phosphate Made of Eggshells
Through A Hydrothermal Method
Name
: Noldy Hutagalung
Student ID
: G74090023
Approved by
Dr Kiagus Dahlan
Supervisor
Dr Irmansyah, MSi
Co Supervisor
Endorsed by
Dr Akhiruddin Maddu, MSi
Head of Physics Department
Date of graduate :
PREFACE
Praise to my Almighty God for His blessing I could finish this research
succesfully. Love and hug to my lovely family for support and pray me all the
time specially to my big mommy and daddy, to my sisters Helga, Maria, Yana,
and Eve, to my brohers Rommy, David, Elnoa for everything that you gave to
make me laugh. My respect and honour to my supervisors : Dr. Kiagus Dahlan,
Dr. Irmansyah, M.Si, and Setia Utami, M.Si for all moment to teach me how to do
this research well. I thank God to have friends like Jise Hari, Margaretha Septiana,
Ratna Sari, Saima Mega, Nasib Pols and PARTARU members for support me all
the time. My big thank to 46 Physics student and biomaterialler : Upri, Indri,
Budi, Kak Aisyah, Mita for helping me while doing this research, then to Pena
stationary : Dede, Vina, Rady and Kania, my dorm mates : Ade, Muga, and Desy.
The author hopes much suggestions to make this research will be better and author
do hope this research will be usefull to everyone.
Bogor, September 2013
Noldy Hutagalung
CONTENTS
LIST OF TABLES
vi
LIST OF FIGURES
vi
LIST OF APPENDICES
vi
INTRODUCTION
1
Background
3
Objectives of Research
3
Purposes of Research
3
Usefulness of Research
3
Scope of Research
3
Hypothesis
3
EXPRIMENTAL METHOD
3
Material and Equipment
3
Preparation of Eggshell
4
Preparation of Sample
4
Hydrothermal Method
5
Sintering Process
6
Sample Characterization by x-ray diffraction (XRD)
7
Sample Characterization by fourier transform infrared (FTIR)
7
RESULT AND DISCUSSION
8
Calcination Process and synthesis of BCP
8
Hydrothermal and Sintering Process
8
Phase, average crystallite size (ACS), and lattice parameter of BCP 9
Phase Groups of BCP
12
Phase of HA with and without Sintering Process
13
CONCLUSION AND SUGGESTIONS
15
Conclution
15
Suggestions
15
REFERENCES
16
APPENDICES
18
CURICULUM VITAE
29
LIST OF TABELS
1 Variation of hydrothermal temperature, sintering temperature and
sintering time
2 Mass of CaO after sintering process
3 Mass of all samples after sintering
4 Ratio of HA and -TCP
5 Parameter lattice of HA
6 Atomic crystall size of all sample
6
8
9
11
12
12
LIST OF FIGURES
1 Preparation then followed by calcination process
2 Solution and suspension preparation followed by stirring and dropping
process
3 Hydrothermal process followed by aging
4 Filtration followed by sintering process
5 XRD patterns of A1-A3 samples with variations of treatment
6 XRD patterns of A4-A6 samples with variations of treatment
7 FTIR spectra of A1-A3 samples with variations of treatment
8 FTIR spectra of A4-A6 samples with variations of treatment
9 XRD patterns of HA1
10 XRD patterns of HA2
4
5
6
7
10
11
13
13
14
15
LIST OF APPENDICES
11 Flowchart
12 Data JCPDS (Join Committee on Powder Diffraction Standards)
13 XRD patterns and FTIR spectra
19
20
22
INTRODUCION
Background
The bone damage is a serious health problem because bone is one of the
most important organs of human. However, bone damage is not only caused by
work accidents but diseases can cause bone damage. Some deseases caused bone
damage are osteoporoses, bone tumors, certain cancer or a brittle bone desease
called osteogenesis.1,2 The bone implantation is the way to treat bone damage and
while doing this we need appropriate materials. The composition of material that
we will use has to similar with bone composition.
In this era, the development of biomaterial not only talked about synthesis
but and characterization but new challanges also. Researchers not only interest
focuses primarily on synthesis and characterization, biological behavior of
material and porouess are important to know. In order to develop material must
have support knowledge of different disciplines and made a colaboration.3 In the
following some interdisiplinaries research that support are biochemistry,
biomedical engineering and medical sciences.
An ideal biomaterials that used for implantation should be has biological
behaviors such as bioactive, biodegradable, non-toxic and biocompatible with
human body.1,2
A biomaterial is a synthetic material used to replace part of living system or
to function in intimate contact with living tissue.3 Biomaterial has been
acknowledged as a bone graft material in a range of medical and dental
applications due to their similar chemical composition with natural bones.
Generally, bone substitution materials such as autograft, allograft and xenograft
are used to solve problems related to bone trauma and fractures. Material which
obtained from the human involved called autograft, from the other human called
allograft and from animal called xenograft.2 But, none of these materials provide a
perfect replacement of the bone due to mechanical and biological instability and
incompatibility. Recently, calcium phosphate bioceramics such as calcium
phosphate, tri-calcium phosphate and hydroxyapatite are identified as most
suitable bone substitution materials to serve the demand. Unlike other calcium
phosphates, HA does not break under physiological conditions. In fact, it is
thermodynamically stable at physiological pH and actively takes part in bone
bonding.4
A biomaterial is an inert material which implanted into living systems to
change living tissue or organ function. The purposes of biomaterials are used to
implant in living sytems’s bone are repair, replace and restore organ or tissue.
Biomaterials that developed widely are hydroxyapatite, -TCP and material
mixtures of HA and -TCP is called biphasic calcium phosphate.
2
phosphate, hydroxyapatite and biphasic calcium phosphate has difference
biological behavior each other.3
Hydroxyapatite is the most commonly used calcium phosphate in the
medical fields or dentistry, as it posseses excellent biocompatibility and
osteoconductive. It has chemical formula Ca10(PO4)6(OH)2and a Ca/P ratio of 1.67.
The density of this material in theoritical is 3.156 g/cm3. The crystalinity of this
material has hexagonal rhombic prisms with the space group being P63/m. Its
units cell contains a complete representation of the apatite crystal, consisting of
Ca2+, PO43-, and OH- groups.2,3 The biological behaviors of this material are bioinert and resorbable. Synthesis of hydroxyapatite can be prepared by using a
variety of method and precursor. The variation of method are wet method and dry
method. Wet method by precipitation by involving the neutral reaction of acid and
alkaline solutions or calcium salts and phosphate salts, dry method by heating,
and hydrothermal method.1
Tricalcium phosphate was used as material to synthesize BCP with HA.
The chemical formula of tricalcium phosphate is Ca3(PO4)2. TCP has four
polymorphs are α, , , and super-α. –TCP is stable up to 1125 °C but above
11β5 °C up to 14γ0 °C α becomes the stable phase. Super-α foms between
14γ0°C and the melting point at 1756 °C. –TCP has the rhombohedral space
group R3c with unit cell a=1.0439 nm, c=3.7375 nm (hexagonal setting) with 21
formula units per hexagonal unit cell. The dissolution rate of –TCP was three
times higher than that of HA. The dissolution rate increased shown like this : HA
< –TCP < α-TCP < TTCP.1,3,5 The biological behaviors of this material are
bioactive, bioresorbable, and biodegradable.
HA and TCP have been used, not only for bone implant substitute materials,
but have also been identified for applicability as drugs delivery, antiseptics, food
supplements, and hypothermia treatment agents. The potential of the material as
anticancer agents has been investigated and it has been found that nanoscale HA is
effective in inhibiting cancer cells growth.6 HA and TCP although they have
similar chemical composition but they different in their biological resorbing
capacity. The mixture material of HA and –TCP is called bihasic calcium
phosphate (BCP). Development of BCP especially with HA and TCP has drawn
considerable attention.7-10 In medical application they have found as bone graft
substitute. The resorbability of BCP ceramics depends on ratio of HA and –TCP,
hight ratio will produce high resorbability.
Some kinds of phosphate have been used to react with CaO of some sources
such as eggshell, fishbone, seashell, etc. Kinds of phospate which generally used
are H3PO4, (NH4)2HPO4, and Na2HPO4.1-12 The eggshell represents the 11% of
the total weight of the egg and is composed by calcium carbonate (94%), calcium
phosphate (1%), orghanic matter (4%) and magnesium carbonate(1%).12,13
3
Objectives of Research
1
2
What the effects of variation of hydrothermal-temperature, hydrothermalholding time and sintering-temperature to produce biphasic calcium
phosphate with ratio of hyrdoxyapatite and -tricalcium phosphate ?
Hows the final result of characterization by using X-ray diffraction (XRD)
and fourier transform infrared (FTIR) ?
Purposes of Research
1
2
3
To synthesize biphasic calcium phosphate using eggshells
To characterize biphasic calcium phosphate by using XRD and FTIR
To get BCP with ratio of HA and -TCP are 60 : 40 and 70 : 30
Usefulness of Research
The useful of this research was to developed biomaterial as bone graft and
mixed two different phase which is called biphasic and obtained new calcium
source from eggshells.
Scope of Research
The aim of this research was to synthesize biphasic calcium phosphate by
hydrothermal method. Calcium source got from eggshells by calcination process
by freeing CO2 and (NH4)2HPO4 as phosphate source. Sample was characterized
by X-ray Diffraction and Fourier Transform Infrared.
Hypothesis
Variation of hydrothermal-temperature, hydrothermal holding time and
sintering temperature can produce BCP with ratio of HA and -TCP are 60 : 40
and 70 : 30. The ratio as mentioned above can produce an ideal material to
implant to human or other living system.
EXPERIMENTAL METHOD
Material and Equipment
The materials which used in this research made of eggshells in chemical
form CaCO3 was calcinated by freeing CO2 with final result of calcination was
CaO as calcium source, (NH4)2HPO4 as phosphate source, aquades (H2O). These
are the equipment : beaker glass, stirrer equipment, furnace, precipitation.
4
equipment, and hydrothermal reactor. Samples was characterized by using X-ray
Difraction and Fourier Transform Infrared.
Preparation of Eggshell
Washing eggshell to eliminate inner membrane of eggshells by using
aquades, then use furnace to calcinate it in 1000°C at heating rate 5°C/minute for
5 hours. At this process eggshel transform from calcium carbonate into calcium
monoxide by freeing carbon dioxide (CO2).14 The material which produce of
calcination process is CaO (calcium monoxide). Sample preparation and
calcination process are shown by the Figure 1 and the chemical reaction of this
process is shown below.
CaCO3 → CaO + CO2
Eggshells putted
into crucible
Calcination T=1000 °C
for 5 hours
CaO after
calcination process
CaO after grinding
Figure 1 Preparation then followed by calcination process
Preparation of Sample
This reasearch starting from suspens of calcium preparation from 0.5 M
CaO was mixed with 100 ml water destilled. Then phosphate solution preparation
from 0.3 M (NH4)2HPO4 was mixed with 100 ml water destilled. Suspens of
5
calcium was stirred by using stirrer equipment at 300 rpm for 20 minutes. At 5
minutes left after stirrer was ran then mix calcium solution and phosphate solution
by using dropping equipment until stirrer stop. Calcium solution, phosphate
suspensions followed by stirring and dropping process are shown by the Figure 2
and the chemical reaction of mixing process is shown bellow :
10CaO + 14H2O + 6(NH4)2HPO4 → Ca10(PO4)6(OH)2 + 18H2O
0.3 M (NH4)2HPO4
+ H2O
0.5 M CaO + H2O
Stirring and
dropping procces
Mixed of calcium suspension
and phosphate solution
Figure 2 Solution and suspension preparation followed by stirring and dropping
process
Hydrothermal Method
The mixed of solution and suspension moved in hydrothermal reactor.
Hydrothermal method is one of methods which considered by researchers to
synthesize calcium monoxide be a biomaterial which has high quality, high
crystalinity and high reactivity. Hydrothermal temperature and sintering
temperature was variated while synthesizing sample. The variation of treatment is
shown in Table 1.
6
Tabel 1 Variation of hydrothermal temperature, sintering temperature, and
sintering time
Sample
code
A1
A2
A3
A4
A5
A6
HA1
HA2
Hydrothermal
temperature
(°C)
250
250
250
300
300
300
150
200
Hydrothermal
rpm (rpm)
300
300
300
300
300
300
300
300
Sintering
temperature
(°C)
800
900
1000
800
900
1000
800
-
Sintering
time
(hours)
5
5
5
5
5
5
5
-
Hydrothermal
pressure (psi)
245
245
245
510
510
510
242
246
The holding time for all samples were 10 hours but special for HA2 was not
sintered and the holding time was 24 hours followed by drying process in 50 ºC
for 8 hours. After all process done then solutions was kept in hydrothermal tube
for overnight or generally this process is called “aging”. Hydrothermal process
and aging are shown by the Figure 3.
Suspension
Hydrothermal
process + aging
Figure 3 Hydrothermal process followed by aging
Sintering Process
Sintering process starting from dropping process. Aging process produced
final solution then solution was filtrated. The residue of solution then moved into
crucible then put in to furnace at 800°C, 900°C, 1000°C at heating rate
5 °C/minute for 5 hours but sample must be dried first at 110 °C with heating rate
5 °C/minute for 5 hours. Filtration and sintering process are shown by the Figure
4.
7
Filtration
Sintering process
Sample after
sintering process
Sample after grinding
process
Figure 4 Filtration followed by sintering process
Sample Characterization by X-ray Diffraction
All samples characterized by using X-ray diffraction GBC EMMA. 2 gr of
each sample putted on holder then setted sample’s name, initial angel 10º, final
angle 80º, CuKα = 1.54056 and analysis speed of computer. The purpose of this
process was to identify crystal structure and lattice parameter of each sample.
Sample Characterization by Fourier Transfrorm Infrared
All samples charcterized by using Fourier Transform Infrared ABB MB
3000. The FTIR spectra used KBr pellet technique. 200 mg KBr mixed with 2 mg
of each sample. The fourier transform infrared spectroscopy employed to
characterize the different functional groups of hydroxyapatite and tricalcium
phosphate. The FTIR used vibration energy of functional groups of HA and TCP
such as PO43-groups, CO32- groups and OH- groups.
8
RESULTS AND DISCUSSION
Calcination Process and synthesis of BCP
The variety of method to synthesize powder are wet and dry method. Wet
method by precipitation by involving the neutral reaction of acid and alkaline
solutions or calcium salts and phosphate salts, dry method by heating, and
hydrothermal method.1 Two kinds of raw material which commonly used to
synthesize BCP are synthetic material and natural material. In this research as
calcium source was waste material it was eggshells.
Calcination was the process to get CaO from CaCO3 by using furnace in
1000 °C at heating rate 5°C/minute for 5 hours called optimum condition of
caltination. At this process eggshells transform from calcium carbonate into
calcium monoxide by freeing carbon dioxide (CO2). Chemical reaction of
calcination process is shown below :
CaCO3 → CaO + CO2
CO2 can be eliminated at higher temperature than 750 ºC, Magnesium (Mg)
can be eliminated in 650 ºC, and organic composition can be eliminated in 80 ºC –
100 ºC.1
The synthesis of BCP produced calcium phosphate ceramics which are
have an important position among other biomaterials, because they are considered
to be almost fully biocompatible with living bodies when replacing the hard bone
tissues, they are hydroxyapatite (Ca10(PO4)6(OH)2) and tricalcium phosphate
(Ca3(PO4)2 ) are currently recognized as ceramic materials that significantly
simulate the mineralogical structure of bone. In this research the molarity ratio of
Ca and P was 0.5 M : 0.3 M because concentration ratio of calcium and phosphate
of HA was 1.67. The reaction of synthesis in hydrothermal reactor as shown
below:
10CaO + 14H2O + 6(NH4)2HPO4 → Ca10(PO4)6(OH)2 + 18H2O
Product of calcination process was CaO and the mass of CaO is shown in Table 2.
Table 2 Mass of CaO after calcination process
Mass of Ca(CO)3 (gr)
21.896
22.722
25.172
22.963
21.666
21.395
20.266
24.716
27.059
27.185
18.954
19.963
Mass of CaO (gr)
11.524
12.083
13.592
12.453
11.453
11.693
10.893
13.294
14.546
14.500
9.914
10.651
9
Hydrothermal and Sintering Process
As mentioned before two kinds of powder synthesis are wet method by
precipitation by involving the neutral reaction of acid and alkaline solutions or
calcium salts and phosphate salts, dry method by heating, and hydrothermal
method. HA prepared by the wet method in atmosphere consist of very small
crystal with lattice deffects. The way to obtain large, perfect, single crystal of HA,
it is preferable to use the hydrothermal technique.3
Hydrothermal synthesis is characterized by the reaction of aqueous
solution in closed recipients under controlled temperature and/or pressure. The
temperature can be elevated above the boiling point of water, reaching the
pressure of vapor saturation. Hydrothermal method is the best method to get
biomaterial high quality, homogenous, high purity, high crystalinity, and high
reactivity. There are three main variables in hydrothermal process they are
temperature, pressure, and chemist potential. The lattice parameter of HA
prepared by the hydrothermal method depend on the reaction temperature and
pressure.1
Sintering was the process in high temperature to appear phase of material
but not all powder synthesises has to passed this process. The use of natural
material such as seashells, eggshells and coral for synthesizing are commonly has
to passed sintering process. Although sample had passed hydrothermal process
with variety of temperature but it had no effect to appeared phase of material
because hydrothermal process just for creating ion groups of HA and -TCP.
Sintering process starting from dropping process. Aging process produced
final solution then solution was filtrated. The residue of solution then moved into
crucible then put in to furnace at 800 °C, 900 °C, 1000 °C at heating rate
5 °C/minute for 5 hours but sample must be dried first to freeing H2O of
suspensions at 110 °C with heating rate 5 °C/minute for 5 hours.
Based on chemical reaction, the result of synthesis was HA but by
sintering with high temperature samples might contain TCP because with
increasing temperature, the apatite structure changes to a new space group and
TCP appears and if in sample contain two different phase it is called biphasic and
in this case two different phase were HA and -TCP which is called BCP. The
mass of all samples are shown in Table 3.
Table 3 Mass of all samples after sintering process
Sample Code
A1
A2
A3
A4
A5
A6
HA1
HA2
Mass (gr)
3.4005
3.3089
3.3330
3.9313
3.7102
3.7103
3.8345
3.3043
10
Phase, Lattice Parameter, and Average Crystallite Size of BCP
The result of XRD characterization for all samples were compared with
the data JCPD (Join Committee on Powder Diffraction Standards). The result of
compared for sample code A1- A6 are shown by the Figure 5 and 6.
Figure 5 XRD patterns of A1 – A3 samples with variations of treatment
Figure 6 XRD patterns of A4 – A6 samples with variations of treatment
11
All figures above show that the result of XRD were same generally. Figure
5 shows the result of sample A1, A2, A3, they were same in hydrothermal
temperature β50 ºC but they are different in sintering temperature. That’s why
their characteristic were different each other. According to thermal stability of
HA and –TCP, with increasing temperature, the apatite structure changes to a
new space group and TCP apprears.3 As mentioned before –TCP is stable up to
11β5°C which is means higher temperature, –TCP appears . Figure 6 shows the
result of sample A4, A5, A6, they were same in hydrothermal temperature at 300
ºC but they were different in sintering temperature.
Calcium phosphate bio ceramics such as calcium phosphate, tricalcium
phosphate and hydroxyapatite has been acknowledged as a bone graft material in
a range of medical and dental applications due to their similar chemical
composition with natural bones. Generally, bone substitution materials such as
autograft, allograft and xenograft are used to solve problems related to bone
trauma and fractures. They are identified as most suitable bone substitution
materials to serve the demand. According to result of evaluated based on data
JCPDS researcher got nothing sample which contained pure HA and –TCP ,
researcher got apatite calcium type A (CAA) and octacal cium phosphate (OCP)
but majority pahses in each sample were HA and –TCP. The existence of OCP
did not matter for ceramics used for implantation because synthetic OCP showed
its osteoconductive characteristics in the bone marrow space such as enhance bone
regeneration at the initial bone apposition stage and stimulate resorption of the
newly formed bone. In addition, biodegrade rate of synthetic OCP is faster than TCP ceramics and OCP enchance bone formation more than HA and -TCP
ceramics when implanted in living system.13
XRD patterns were used to find phase of material, parameter lattice and
atomic crystal size. XRD patterns were compared with data JCDPS and based on
result of XRD patterns above we knew that they are different each other specially
for ratio of HA and –TCP. The strongest peak of all samples is appears at 31.7º
and it is belong to HA. The result of calculated are shown in Table 4.
Table 4 Ratio of HA and –TCP
Sample Code
A1
A2
A3
A4
A5
A6
HA : –TCP
64: 36
66 : 34
62 : 38
74 : 26
63 : 37
63 : 37
The result of comparing XRD patterns and data JCPDS were used to
calculated parameter lattice. The crystallite size of all samples based on full width
half maximum (FWHM) of X-ray diffraction patterns were calculated by using
12
MDI Jade 6 program. The lattice parameter and average crystallite size are shown
in Table 5 and 6.
Table 5 Lattice parameter of HA
Sample code
A1
A2
A3
A4
A5
A6
HA1
HA2
a (Å)
10.056
9.519
9.741
9.427
11.738
9.407
9.414
9.468
Table 6 Average crystallite size of all samples
Sample code
A1
A2
A3
A4
A5
A6
HA1
HA2
c (Å)
7.352
6.959
7.127
6.893
8.510
6.881
6.878
6.925
ACS (nm)
42.890
49.562
54.298
49.180
40.340
53.810
49.332
34.911
The lattice constant of HA prepared by the hydrothermal method depend
on the reaction temperature and pressure.1 According to the data that we got while
synthesizing BCP higher temperature hydrothermal, higher pressure. The lattice
parameters of the synthesized BCP samples were obtained from the diffraction
pattern by fitting the peaks of identified reflections. Generally crystallite size of
all samples were similar except HA2 because optimum phase of hydroxyapatite
would be appeared in high temperature because CaO that we used in this synthesis
was wastes material.
Phase Groups of BCP
Previous report clearly demonstrated that infrared spectroscopy is an
indispensable tool in order to learn some special features of the preparation of
calcium hydroxyapatite ceramics.10 Therefore, Fourier Transform Infra red
characterization was carried out for BCP samples to study the spectral
characteristics which indicated the chemical bonding. The result of FTIR
characterization for all sample are shown by the Figure 7 and 8.
13
Figure 7 FTIR spectra of A1 – A3 samples with variation of treatment
Figure 8 FTIR spectra of A4 – A6 samples with variation of treatment
14
The spectra indicates the formation of a typical HA structure containing
PO43- band and OH- band, TCP structure containing PO43- band, and OCP
structure containing HPO42- band. Phosphate characteristic absorption bands
appear at wavenumber 650 - 550 cm-1 is called v4, 1200 – 900 cm-1 is called v3,
430 – 460 cm-1 is called v2 and around 960 cm-1 is called v1. Carbonate absorption
bands between 1600 – 1400 cm-1 is called v1 and around 870 cm-1 is called v2.
Absorption bands around 632 cm-1, 3573 cm-1, and 3649 cm-1 show the OH- group.
Absorption band around 651 cm-1 and 3445 cm-1 show H2O that absorbed by
sample.1,15,16
Phase of HA with and without Sintering Process
In this research two different samples were addded to synthesize as shown
in Table 1 which are different in treatment of variation. For HA1 was
hydroxyapatite with temperature of hydrothermal is 150 ºC, hydrothermal holding
time is 3 hours, and sintered in 800 ºC for 5 hours. Another one was
hydroxyapatite with temperature of hydrothermal 200 ºC, hydrothermal holding
time 24 hours without sintering process (HA2). XRD patterns were compared
with the data JCPDS. The result of these synthesises were different each other
specially for peak strong. HA1 had a stronger peak of all peak in this sample at
31.7° with intensity at 385 counts/s. HA2 had stronger peak at 31.7º with intensity
at 267 counts/s. The result of compared for two samples hydroxyapatite are
shown by the Figure 9 and 10.
Figure 9 XRD patterns of HA1
15
Figure 10 XRD patterns of HA2
Figure 10 shows the sample which totally different from those sample
which sinterred before. The main reason is because HA phase by using wastes
material such as eggshells, seashells and others wastes material will be created if
we sintered in high temperature. When hydrothermal process was run sample
synthesized in 250 ºC for 24 hours and analyzed result by using database JCPDS
we got OCP, HA, and -TCP and majority phase is HA. But if we compared result
analys of HA2 and sample with sintering process, sample with sintering better
than HA2 because the crysstalline structure of this sample was amorphous as we
see in Table 6. Otherwise if we use calcium nitrate tetrahydrate (Ca(NO3)2.4H2O)
or other synthetic materials the synthesized did not need sintering process.10
16
CONCLUSION AND SUGGESTIONS
Conclusion
This research was to synthesize BCP made of eggshells through a
hydrothermal method. Biphasic calcium phosphate was mixed of HA and -TCP.
Calcium source was from waste materials it was hen’s eggshells. The chemical
form of eggshells is CaCO3. Calcination was the process to get CaO by freeing
CO2 from CaCO3 using furnace at 1000 °C with heating rate 5 °C/minute for 5
hours. Each sample was mixed of calcium suspension and phosphate solution.
Ratio molar of Ca and P was 0.5 M : 0.3 M to synthesize BCP. Hydrothermal was
the method to synthesize calcium monoxide be a biomaterial high quality, high
crystalinity and high reactivity. Hydrothermal temperature and sintering
temperature were variated. For sintering process higher temperature, tricalcium
phosphate appears and for hydrothermal process higher treatment of temperature,
higher pressure in hydrothermal tube. Three main variables in hydrothermal
process were temperature, pressure, and chemist potential.
Product of sintering process was characterized by using XRD and FTIR.
XRD patterns were compared with the data JCPDS to know what the phase of
product. The result of comparing process was majority phases were HA and TCP. The lattice parameter and average cryztallite size were calculated by
comparing XRD pattern with database JCPDS. Higher treatment sintering
temperature had effect on atomic crystal size of BCP. To support the calculated
result of characterized by using XRD we got functional groups of HA and TCPas PO43-groups, CO32- groups and OH- groups of FTIR characterized.
Suggestions
1. As mentioned before with increasing temperature, the apatite structure
changes to a new space group and TCP apprears that’s why we suggest touse
sintering temperature higher than 1000 ºC to get more various of ratio of HA
and -TCP.
2. In this research all samples were characterized by using XRD and FTIR, the
author recomend to use SEM to know about the morphologh structure of BCP.
3. To get sample without impurities the residue of filtration must be washed by
using water destilled.
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18
APPENDICES
19
Appendix 1 Flowchart
Materials and equipment preparation
Ready
No
Sample preparation
Hydrothermal synthesis
CaO, (NH4)2HPO4, H2O
Sintering
Biphasic calcium phospate
FTIR and XRD characterization
Data analysis
Report arrangement
Figure A7 Steps which author did as basic principle to research
20
Appendix 2 Data JCPDS of (a) -TCP, (b) HA, (c) OCP and (d) CAA
(a)
(b)
21
(c)
(d)
22
Appendix 3 XRD pattern of sample A1
XRD pattern of sample A2
23
XRD pattern of sample A3
XRD pattern of sample A4
24
XRD pattern of sample A5
XRD pattern of sample A6
25
XRD pattern of sample HA1
XRD pattern of sample HA2
26
FTIR spectra of sample
FTIR spectra of sample A2
27
FTIR spectra of sample A3
FTIR spectra of sample A4
FTIR spectra of sample A4
28
FTIR spectra of sample A5
FTIR spectra of sample A6
29
CURICULUM VITAE
The author was born in Sipoholon on December 23rd 1991 as fifth child of
eight from merried of Jodi Hutagalung and Marlince Sinaga. The writer was
educated at Elementary school SDN 0178492 in 1997-2003, Junior High school at
SMPN 1 Sipoholon in 2003-2006, Senior High School at SMAN 1 Sipoholon in
2006-2009. The author was accepted at Bogor Agricultural University in 2009 by
means of USMI (Ujian Seleksi Masuk IPB) in departmentof Physics . First year
the writer passed Tingkat Persiapan Bersama (TPB) then second year the writer
studied as student in department of Physics. The writer was one of laboratory
assistant of 1st Physics experiment.