Synthesis of Hydroxyapatite from egg shell and preparation ... · Ahmed M. Saeed [6] have worked on synthesis of calcium hydroxyapatite powder from hen’s eggshell and orthophosphoric

International Journal of Advanced Information Science and Technology (IJAIST) ISSN: 2319:2682 Vol.5, No.8, August 2016 DOI:10.15693/ijaist/2016.v5i8.36-47

36

Synthesis of Hydroxyapatite from egg shell and preparation of bone like Bio-composites

using it Prof. S.L.Pandharipande Associate Professor, Department ofChemical

Engineering, L.I.T, RTMNU, Nagpur, India

Miss. Smita S. Sondawale M.tech Chemical Engineering student,

L.I.T, RTMNU, Nagpur, India

Abstract – Hydroxyapatite (HAp) is an inorganic

constituent of human bone. Human bone contains

67% mineral matter, 22% collagen, 9% water and

remaining proteins, carbohydrates, lipoids, etc. Bio-

composites that would resemble natural bone

containing HAp as inorganic matter blended with

organic biomaterials are being researched. The

present work is aimed at synthesis of HAp from egg

shell, silica from rice husk and bio-composites C1 to

C6 using varying compositions of HAp, silica,

chitosan, carboxymethyl cellulose (CMC) & gelatin.

Based on the Fourier Transform Infrared

Spectroscopy (FTIR) analysis it can be said that HAp

and silica are successfully synthesized. The bio-

composite of HAp with CMC & HAp with CMC,

chitosan, gelatin & silica are tested for their

mechanical strength such as hardness & compressive

strength. The composite of HAp with CMC &

chitosan are tested for their antimicrobial property.

The test results show that the composites have

comparable mechanical properties to that of

cancellous bone of human and also good

antimicrobial property.

Index terms – Hydroxyapatite, Bone like bio-composites, Egg shell, Compressive strength, Hardness, antimicrobial property.

I. INTRODUCTION

Hydroxyapatite (HAp), Ca10(PO4)6(OH)2 is

an important inorganic biomaterial consisting of

calcium and phosphorous which are important

minerals found in human bone. Human bone

constitutes 67% mineral matter comprising HAp,

22 % organic matter which is mainly collagen, 9%

water and remaining is proteins, carbohydrates,

polysaccharides etc. Thus, natural bone is a

composite of inorganic and organic matrix. It

consists of calcium and phosphorous in the ratio

1.67 which is same as the stoichiometric ratio

present in the molecular formula of HAp [1],[2].

HAp provides sufficient toughness & hardness

whereas the organic matrix provides the

mechanical strength such as compressive & tensile

strength to bone. Hence, composites of HAp with

certain organic polymers that resembles collagen

chemically can be used for bone grafting. The

organic part may be substituted by polymers such

as Chitosan, Gelatin, Carboxymethyl Cellulose

(CMC), etc.

HAp can be synthesized effectively from egg

shell. The idea is to minimize the egg waste

accumulation caused by the day to day increase in

consumption of eggs, due to the large demand of

the poultry products such as cakes, fast food, etc.

The disposal of egg shell waste in land may cause

land degradation that can be minimized by

effective utilization of egg shell in HAp synthesis.

Also silica can be synthesized from rice husk. Rice

husk is a waste product of rice mill and can lead to

waste accumulation if not disposed of properly. It

contains 98% silica. Thus can be utilized in the

synthesis of silica.

The present work aims to synthesize

biomaterials; HAp using egg shell, Silica using rice

husk and bio-composites of HAp using different

organic biomaterials mentioned.

II. LITERATURE REVIEW

i) Synthesis of Hydroxyapatite

Eric M. Rivera et al [3] have worked on

synthesis of hydroxyapatite from eggshell. In this

paper HAp was synthesized from egg shell by

thermally treating the egg shells The Calcium

Oxide thus obtained was transformed into HAp in a

phosphate solution. The Ca/P ratio of 1.67 was

used. The final product is characterized by X-ray

diffraction and scanning electron microscopy SEM. Gre´ta Gergely [4] have worked on preparation

and characterization of hydroxyapatite from

eggshell and phosphoric acid as raw materials. The

present work aimed to synthesize HAp via

mechano chemical activation using ball milling and

attrition milling and the phase stability and powder

characteristics were studied. The structures of the

HAp were characterized by X-ray diffraction, scanning electron microscopy and infrared

spectroscopy. P. Hui [5] have worked on synthesis of

hydroxyapatite bio-ceramic powder by

hydrothermal method. Egg shells were treated

thermally for and the calcium oxide obtained was

treated with Tricalcium Phosphate solution. The

final product was characterized by X-ray


37

diffraction, Scanning electron microscopy (SEM),

Fourier Transform Infrared Spectroscopy (FT-IR), thermal analysis (TG–DTA). Ahmed M. Saeed [6] have worked on

synthesis of calcium hydroxyapatite powder from

hen’s eggshell and orthophosphoric acid. X-Ray

diffraction (XRD) and Fourier Transform Infrared

(FT-IR) technique was used to investigate the

formation of HAP powder.

ii) Synthesis of Sillica

K.V. Selvakumar [8] have worked on

extraction of Silica from burnt paddy husk.

Demineralization of rice husk was done using

sulphuric acid or nitric acid or hydrochloric acid.

Then the residue was treated with NaOH and the

filtrate so obtained was titrated with acid to obtain

the precipitate of silica. The effect of different

concentration of ascid and base on the yield of

silica yield of Silica was examined. The product

was analyzed using SEM.

Majid Monshizadeh [9] carried out the same

procedure as above and used magnetic stirrer for

continuous stirring of the solution. Silica obtained

was characterized using FTIR, XRD and SEM

analysis.

iii) Synthesis of Bio-composites

Subhadra Garai [10] worked on the

synthesis of bio-composite of HAp with CMC.

The process consisted of mixing of polymeric

solution of CMC with the calcium nitrate tetra

hydrate solution and then the solution mixture was

treated with di-ammonium hydrogen phosphate

solution. Varying wt% ratio of CMC and HAp

were used. The slurry was then dried in Teflon

molds at 500C for 72hrs. The bio-composite was

tested by using FTIR, SEM, XRD and

compressive testing.

Edgar B. Montufar [11] synthesized HAp-

Gelatin composite scaffolds by mixing α –

tricalcium phosphate (α- TCP) with HAp and

the Liquid phase of gelatin was prepared adding

Na2HPO4 as an accelerant. Both the liquid and

the powder phase were mixed and were molded or

placed into the syringe at room temperature. The

characterization were carried out before and after

immersion of the composites in Ringer’s Solution

i.e; 0.9 wt% of NaCl solution and 100 %

humidity, using FTIR, SEM, XRD and

compressive testing.

Mehdi Kazemzadeh Narbat [12]

synthesized HAp – Gelatin (GEL) composite with

30, 40 & 50 wt % HAp composition. The

composites were prepared by directly mixing HAp

powder and the gelatin solution. The composites

were then molded into Teflon molds and

Characterization was done using FTIR, SEM and

mechanical testing.

III. PRESENT WORK

A. Methodology

The schematic of the process followed in the

present work [7] is given in figure 1.

B. Materials

Glassware, egg shell, Diammonium Hydrogen

Phosphate (DAP), Chitosan, Gelatin,

Carboxymethyl Cellulose CMC, rice husk.

C. Method

The present work is divided 3 parts:

Synthesis of Hydroxyapatite from Egg

shell

Synthesis of Silica from Rice husk

Synthesis of bio-composites

i) Synthesis of HAp using Egg shell

The outline of the procedure [3-7] is as given

below. The pictorial representation of the process is

given in fig 2.

Cleaned and boiled egg shells were treated

thermally at 700-8000C for 4 hrs and

crushed to obtain CaO powder.

Samples of 0.6M DAP solution and 0.3M

of CaO solution were prepared and mixed

in varying proportions thoroughly for 1 hr

under continuous boiling.

The precipitate of HAp obtained was

filtered using whatmann filter paper and

dried in oven at 800C for 4-5 hrs.

The final product was sintered in furnace

at 700-8000C for 1 hr.

Three samples of HAp; P1, P2 &P3 are

synthesized having same as

stoichiometric, half and quarter quantity of

DAP respectively.

ii) Synthesis of silica using rice husk

The outline of the procedure [8],[9] followed is as

given below and the pictorial representation of the

process is given in fig 2.

Rice husk was demineralized by boiling in

1N HCl solution followed by heating for

2hrs .

Demineralized rice husk was washed till

neutral pH of washed water obtained.

Washed rice husk was then treated with

2.5 N NaOH for 2hrs at 80-1000C.

The residue obtained was discarded & the

filtrate was titrated with 1N HCl solution

The above solution was left to age for

24hrs at ambient conditions.


38

The precipitate of Silica obtained was

filtered, oven dried and sintered in

furnace.

Two runs were carried out to prepare 2

samples of silica.

iii) Preparation of Bio-composites

The outline of the procedure followed is as given

below:

The preparation of [10-12] bio-composites

consists of mixing of organic binders;

Chitosan, Gelatin, Carboxymethyl

Cellulose and inorganic components;

Hydroxyapatite and Silica .

The Organic binders were prepared in gel

form by adding sufficient amount of

water.

Hydroxyapatite was added to the gel of

organic binders and stirred till

homogeneous mixture was formed.

The mixture was poured into cubical

molds and kept in deep freezer followed

by air drying for at least 3 days.

The further details of the composite in

terms of its composition is given in table

4.

The following 6 types of composites are

prepared:

Composite 1: C1: CMC + P1



Composite 4: C4: CMC + HAp + Chitosan

+ Gelatin + Silica

Composite 5: C5: CMC + HAp

Composite 6: C6: CMC + HAp + Chitosan

Organic

binders

Gelatin Chitosan CMC

Egg

Shell

DAp

HA p

(P1)

Stoichiometric DAP (1)

Half stoichiometric (2)

Quarter Stoichiometric (3)

Hydrothermal

method

HAp

(P2)

HAp

(P3)

(1) (2) (3)

Analysis

FTIR

SEMC1

C2

C3

C5

C4

C4 Silica

Analysis

Rice Husk

Chemical

treatment

FTIR

SEM

Mechanical testing

Compressive test

Hardness test

Figure 1: Block diagram of the process followed

Egg Shell Boiling Drying in oven Dried egg shell Sintering


39

0.6M DAP solution 0.3 M CaO solution CaO powder

0.6M DAP Heating in oven Precipitate

+ 0.3 M CaO of HAp

Product HAp Sintering Air dried precipitate Filtration

Figure 2: Pictorial representation of HAp synthesis

Rice Husk HCl treatment Filtration NaOH treatment Filtration

Silica gel Filtration Acid treatment

Figure 3 : Pictorial representation of Silica Synthesis

(a) (b) (c) (d) (e) (f)

Figure 4 : Actual photographs of bio-composites synthesized a) C1 b) C2 c) C3 d) C4 e) C5 f) C6

IV. OBSERVATIONS

The details of the process parameters & the yield

obtained in the synthesis of HAp, silica and bio-

composites are given in tables 1, 2, 3 & 4

respectively

S

r

n

o

Samp-

le

name

Wt of

egg

shell

(gm)

Tempera-

ture of

calcinatio-

n

(0C)

Time of

calcinati-

on

(hrs)

CaO

formed

(gm)

Water

added

to CaO

(ml)

DAP

take-

n

(gm)

Wat-

er

adde-

d to

DAP

(ml)

Ca/

P

rati-

o

Wt of

Pro-

duct

(gm)

1 H-1 10 800-850 4 5 530 13 163 1.18 5.97

HS-1 The product H-1 was sintered at 800-8500C for 1hr


40

2 H-2 15 800-850 4 8 476 15 189 1.65 4


3 H-3 15 800-850 4 8 476 15 189 1.65 4


Table 1: Details of parameters for Synthesis of Hydroxyapatite

Sr no Sample name CaO

(gm)

Water added to CaO

(ml)

DAP Water added to DAP

(ml)

Ca/P ratio

1 P1 4 238 7.5 94.70 1.65

2 P2 4 238 3.75 47.35 3.29

3 P3 4 238 1.875 23.68 6.59 Table 2: Details of parameters for synthesis of HAp by varying Ca/P ratio

Sr no Samp

-le

name

Wt of

RH

(gm)

Vol &

normality

of HCl

solution

(ml)

Hrs of

HCl

treat-

ment

Vol &

molarity

of NaOH

taken

Hrs of

NaOH

treatmen

-t

Vol of

filtrat

-e

(Na2Si

O3)

(ml)

Vol

of

HCl

solu

-tion

add-

ed

(ml)

Hrs

of

treat-

ment

Wt

of

Sili-

ca

(gm

)

1 S-1 10 150ml 1N 24hrs 160ml

2.5M

24 275ml 683 24 1.5

SS-1 The product S-1formed was sintered at 800-8500C for 1hr

2 S-2 10 450ml 1N Boile-

d for

2hrs

180ml

2.5N

2 ---- 130 24 0.7

SS-2 The product S-2 formed was sintered at 800-8500C for 1hr

Table 3: Details of parameters for Synthesis of Silica

Sr no Composite name HAp

(gm)

Chitosan

(gm)

CMC

(gm)

Gelatin

(gm)

Silica

(gm)

1 C1 2.37 - 0.26 - -

2 C2 2.37 - 0.26 - -

3 C3 2.37 - 0.26 -

4 C4 4 0.25 0.6 0.25 1

5 C5 4 - 1 - -

6 C6 4 0.3 0.7

Table 4: Details of the composition of bio-composites (weight basis)

V. CHARACTERIZATION

The characterization is divided into two parts:

Characterization of biomaterials

Characterization of bio-composites

A. Characterization of Biomaterials

The biomaterials synthesized i.e; HS-1, H-2,

HS-2, P-1, P-2, P-3, S-1 & SS-2 were characterized

using FTIR and SEM techniques.

i) FTIR analysis

The FTIR analysis of the samples was done for

determination of the various functional groups

present that include OH-, PO4

3- in case of HAp and

Silinol OH group in case of silica. The fig no 5a,

5b, 5c, 5d, 5e, 5f, 5g & 5h show the graphs of FTIR

analysis of HS-1, H-2, HS-2, P-1, P-2, P-3, S-1 &

SS-1 samples respectively.

.


41

(a) (b)

(c ) (d)

(e) (f)

(g) (h)

Figure 5: FTIR analysis of biomaterials synthesized a) HS-1 b) H-2 c) HS-2 d) P1 e) P2 f) P3 g) S-1 h) SS-1

ii) SEM analysis

The SEM analysis of HAp as well as silica

samples was carried out to determine the surface

morphology, approximate particle size and surface

texture. The fig nos 6a, 6b, 6c & 6d shows the

SEM micrographs of HS-1, H-2, HS-2 & SS-1

respectively

.


42

(a) (b)

( c) (d)

Figure 6: SEM micrograph of the biomaterials synthesized a) HS-1 b) H-2 c) HS-2 d) SS-1

B. Characterization of Bio-composites

The bio-composites were tested for

compressive strength, hardness and anti-microbial

properties.

i) Compressive strength testing

The specimen was compressed between two

square plates of 5cm x 5cm dimension with a

crosshead speed of 1mm/min. Fig 7a & 7b show

the load vs displacement characteristics of sample

C4 & C5 respectively. The details of dimensions

of the cubical slabs along with the details of

mechanical compressive strength is given in table

5.

(a) (b)

Figure 7: Load vs Displacement characteristics of bio-composites a) C4 b) C5


43

Sample

no

Bio-composite

Name

Cross Sectional

area (mm2)

Maximum load

(KN)

Strain

(%)

Compressive

strengths (MPa)

Initial Final

1 C4 86.47 - 0.142 41.39 1.642

2 C5 85.42 57.86 2.435 38.55 28.503 Table 5 Compressive strength of composites

ii) Hardness or Fracturability test

The samples C1, C2, C3 are tested for hardness

or fracturability using cylindrical flat probe of 2

mm diameter. The penetration speed of the probe

during testing was 1mm/sec and was penetrated to

a distance of about 4 mm. Fig no 8a, 8b & 8c show

the force vs time characteristic of bio-composites

C1, C2 & C3 respectively. The table 3.6 shows the

hardness of the samples.

(a) (b)

Table 6: Hardness of bio-composites

(C)

Figure 8: Force vs time characteristic of bio-composite

a) C1 b) C2 c) C3

iii) Anti microbial property testing

HAp is an inorganic biomaterial having

strong resistance to microbial growth. The

incorporation of organic binders into the inorganic

HAp must not alter its antimicrobial properties.

The bio-composite should exhibit similar

antimicrobial property. Hence, bio-composite C6

having HAp + Chitosan + CMC, is tested in Blood

agar media for its anti microbial property over

48hrs.

VI. RESULTS & DISCUSSION

The following sections give the detailed

discussion on various observations related to

synthesis & analysis of bio-composite materials as

Force (g )

Time (sec)

1F1D

Force (g )

Time (sec)

1F1D

Sample

no

Bio-

composite

Name

Cross

sectional

area of

probe

(m2)

Maximum

load

(KN)

Hardness

(GPa)

1 C1 3.14 x

10-6

62.24 19.8

2 C2 3.14 x

10-6

55.53 17.7

3 C3 3.14 x

10-6

55.01 17.5

Force (g )

Time (sec)

1F1D


44

presented in the previous section. The discussion is

presented in 5 parts: Synthesis, FTIR, SEM,

compressive, hardness and antimicrobial property

test.

A. Synthesis results of HAp and Silica

The present work has successfully synthesized

HAp and Silica. The process parameters were

chosen at optimal condition based on literature

review. The yield of CaO from egg shell is around

50%. In Run-1 the Ca/P ratio was less as compared

to Run-2 and 3. In Run-2 and 3 the Ca/P ratio is

according to stoichiometry and any undissolved

Ca(OH)2 is discarded. The physical observations of

HAp synthesized is indicative of its formation

based on color, size and texture.

Similarly the yield of silica synthesized varies

from 7-15% and its formation is visually supported

by its white color, crystalline fine powder structure

and texture.

B. Interpretation of FTIR graphs

The interpretation of FTIR graphs is necessary

to confirm the presence of various functional

groups. This is usually done by mapping the graphs

of FTIR of samples with those of standard

functional group as reported in literature. The

detailed interpretation of FTIR graphs of 4 samples

of HS-1, H-2 & HS-2, and S-1 & SS-2 is given in

table 7 and 8 respectively.

As seen from the table 7, the FTIR analysis of

three HAp samples are confirmatory of presence of

all the functional groups. The peak for CO32-

is

seen additionally due to the substitution of either

OH- or PO4

3- by the CO3

2- group during the

atmospheric contact of the product. The FTIR

graphs of the samples P1, P2, P3 show the peaks

for all the functional groups. The difference in the

depth of the CO32-

peak. The depth is seen

increasing with increase in the Ca/P ratio i.e;

decrease in the phosphate content. The reason may

be due to increase in the possibility of substitution

of CO32-

group in place of OH- or PO4

3- [1].

The FTIR analysis of silica samples i.e; S-1

and SS-1 show that the non sintered silica (S-1)

shows the peak for silinol OH present. This may be

due to the presence of absorbed H2O molecules

which disappears in the sintered silica sample (SS-

1). The peaks for the Si-O-Si bond and vibration of

the gel network are seen in both the samples which

constitutes the structural formula of silica [8], [9].

Sr

no

Sample

name

Vibrational frequencies (cm-1

)

PO43-

OH-

CO3

2-

Stretching Bending

Symmetric Asymmetric

1 HS-1 963.04 1019.57 675.73,

600.40

3398.41,

1632.44

2358.82,

1414.32

2 H-2 962.33,

450.97

1022.80 600.49,

559.62

3638.4,

1652.06

3242.02

1418.10,

873.52,

2881.81,

2161.61

3 HS-2 961.79,

469.19

1087.00,

1022.24

562.94,

599.21,

629.39

3640.54,

3571.55,

1652.90

1418.28

4

P-1 962.18,

462.25

1022.32 601.38,

559.90,

500.50

3214.96,

1641.54

2882.90,

2159.87,

1422.64

5 P-2 962.51,

454.56

1022.53 600.22,

559.84

3208.46 2883.44,

2517.08,

2159.91,

1413.32

6 P-3 962.51,

469.79

1025.75 600.79,

561.98

3696.32,

3638.08,

3225.52

2881.27,

2506.01,

2159.81,

1410.22 Table 7: FTIR interpretation of HAp samples


45

Sr

no

Sample

name

Vibrational frequencies (cm-1

)

Silinol OH group Siloxane bonds (Si-O-Si) Vibration mode of gel

network

1 S-1 3750.35, 3744.24,

3734.78, 3710.84,

3689.01, 3675.12,

3628.58, 3618.71,

3587.29, 3566.68

1052.24, 1497.05, 1489.30, 1473.07,

1450.92, 1436.32, 1418.72

795.51, 668.24

2 SS-1 NA 1063.52, 1448.40 787.19

Table 8: FTIR interpretation of Silica samples

C. SEM analysis

The interpretation based on SEM micrographs can

be summarized as follows:

Sample HS-1 show agglomerate formation

with pores not necessarily interconnected.

There is indication of inhomogeneity in

the surface morphology of the sample.

Non-sintered sample H-2 as well as

sintered sample HS-2 show a micro sized

grain agglomerates with interconnected

structures. The size may be ranging from

2-5 micro meter or even less than that.

HS-2 sample is observed to be more

uniform in size and structured particles are

formed. Whereas non sintered sample has

varied nature of size and shape of

particles.

The sintered silica sample i.e; SS-1 has

needle shaped crystalline particles having

size of 10 micro meter in length and upto

1 micrometer in diameter.

D. Compressive strength testing

The interpretation of compressive strength of

composites as mentioned in table 3 is given below:

The thickness reduction of various

samples that can be expressed as % strain

ranged between 38-40 % and. The %

strain is defined as (original thickness-

Final thickness)/Original thickness x 100.

The composite formed by addition of

silica to HAp, CMC, Chitosan and Gelatin

showed compressive strength of 1.642

MPa. This is least amongst the two and

may be due to non homogeneity of

composite due to the addition of silica.

The compressive strength of sample C5

comprising of HAp and CMC is 28.503

MPa.

The purpose of addition of CMC,

Chitosan and gelatin to HAp is to make it

more like a natural bone containing 67%

inorganic matter and remaining organic

matter. Also they act as binders. The

combination of these have given the best

composites in the form of C5.

E. Hardness and Fracturability test

The hardness of the samples C1, C2 & C3

composed of HAp and CMC in the ratio 90:10 are

in the range of 17-19 GPa The hardness is more

than that of the hardness of cancellous bone ; 0.345

GPa and that of cortical bone; 0.396 GPa [13].

F. Microbial testing

The application of composites synthesized in

present work are in biomedical field and

particularly in bone grafting. The anti microbial

properties of these bio-composites is thus

important. Based on the test report of sample C5

comprised of HAp, Chitosan & CMC, it can be said

that it posess anti microbial properties and no

growth of micro organisms was observed for 48 hrs

[14].

VII. CONCLUSION

The present work addresses to synthesis of

bio-composites of HAp, CMC, gelatin, silica &

chitosan. The main aim of the present work is to

synthesize bio-composites that resemble natural

bone. Natural bone constitutes inorganic to organic

matter in the ratio of 70 : 30. In the present work

HAp is used as an inorganic biomaterial source

whereas CMC, gelatin & chitosan are tried as

organic biomaterial source. The methodology of

the present work includes synthesis of HAp and

silica and their bio-composites using mentioned

biomaterials.

Based on the FTIR analysis of HAp and silica

samples, it can be said that these biomaterials are

successfully been synthesized. Three sample of

bio-composites C1, C2 & C3 having 90:10

composition (wt%) of HAp and CMC are

synthesized. Similarly C5 is synthesized using

same biomaterials with 80:20 composition (wt%).

C6 contains HAp, CMC & chitosan .having


46

inorganic to organic ratio of 80:20. C4 contains 5

biomaterials; HAp, silica, chitosan, CMC &

gelatin.

The composites C1, C2 & C3 are tested for

hardness test and its value is in the range of 19-17

GPa, greater than the hardness of cancellous and

cortical bone of human. The sample C4 & C5 are

tested for compressive strength. The compressive

strength of sample C4 is 1.642 MPa and that of C5

is 28.503 MPa. The strength of C4 is less as

compared to cancellous bone which is in the range

of 2-20MPa. The reason may be due to insufficient

amount of organic matter to bind the inorganic

components i.e; HAp & silica. The strength of

composite C5 is greater than that of cancellous

bone. Composite C6 is tested for its antimicrobial

property and it was found that there was no

microbial growth within 48 hrs.

Thus, it can be concluded that the present work

has successfully synthesized bone like bio-

composites using HAp, CMC, gelatin, silica &

chitosan which has comparable mechanical as well

as antimicrobial properties to that of human bone.

VIII. ACKNOWLEDGEMENT

The authors are thankful to Director L.I.T,

RTMNU, Nagpur, for the facilities and

encouragement provided.

The authors are thankful to H.O.D, Food

technology department, L.I.T, RTMNU,

Nagpur, for hardness test.

The authors are thankful to H.O.D,

Physics department, University campus,

Nagpur, for SEM analysis

The authors are thankful to H.O.D,

Chemical and Metallurgical department,

V.N.I.T, Nagpur, for FTIR analysis and

compressive test.

The authors are thankful to Parate

Pathology laboratory for Antimicrobial

test.

The authors are thankful to India mart for

providing DAP samples.

IX. REFERENCES

1. Shekhar L. Pandharipande, Smita S.

Sondawale, Review on the characterization

methods of Hydroxyapatite and its Bio-

composites, International Journal of Science,

Engineering and Technology Research

(IJSETR) Volume 5, Issue 7 (2016).

2. Anton Ficai, Ecaterina Andronescu, Georgeta

Voicu and Denisa Ficai (2011). Advances in

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47

Author’s Profile

Shekhar Pandharipande is an Associate Professor in Chemical Engineering Department of Laxminarayan Institute of

Technology, Rashtrasant Tukadoji Maharaj University, Nagpur. He did his masters in 1985 & joined LIT as a Lecturer. He has coauthored three books titled ‘Process Calculations’, ‘Pririnciples of Distillation’ & ‘Artificial Neural Network’. He has two copyrights ‘elite-ANN’ & ‘elite-GA’ to his

credit as coworker and has more than 60 papers published in journals of repute.

Smita Sondawale is a M.tech (Chemical

Engineering) student from Laxminarayan Institute

of Technology, Nagpur.

Documents

Synthesis of Hydroxyapatite from egg shell and preparation ... · Ahmed M. Saeed [6] have worked on synthesis of calcium hydroxyapatite powder from hen’s eggshell and orthophosphoric