UNIVERSITY OF YANGON

DEPARTMENT OF CHEMISTRY

Preparation and Characterization of Calcium Carbonate as

Bio-sorbent from Waste Eggshells for Removal of Heavy
Metals

by
Chit Su San Mg
2 MSc- BC-19

Date of Seminar : 30.5.2020

 UNIVERSITY OF YANGON
DEPARTMENT OF CHEMISTRY

Date : 30.5.2020

Title : Preparation and characterization of Calcium Carbonate as

Bio-sorbent from Waste Eggshells for Removal of Heavy Metals

Candidate : Ma Chit Su San Maung (2MSc-BC-19)

Supervisor : Dr Yi Yi Myint ,Assistant Lecturer

Department of Chemistry, University of Yangon
 Abstract

 PCC was prepared from chicken eggshell powder (ESP) by dissolving 100g ESP in 1M
hydrochloric acid at room temperature .

 Under vigorous stirring, PCC was precipitated from the ESP –dissolving solution
using 1M sodium carbonate solution .

 The dried PESP (precipitated eggshell powder) was then characterized using
scanning electron microscopy (SEM),X-ray diffraction (XRD),UV-Visible and FT IR
Spectroscopy, and Thermo gravimetric and Differential Thermal Analyzer (TG -DTA ).
The adsorption of Cu(II)nitrate onto eggshell was investigated .

 The effects of contact time , initial pH of solution ,adsorbent dosage.

 RESEARCH OUTLINE

 Aim and Objectives

 Introduction

 Materials and Methods

 Results and Discussion

 Conclusion

 References
 AIM

To study the heavy metals (Cu2+, Ni2+ and Cr3+ ions) removal efficiency from aqueous
medium by using calcium carbonate as bio-sorbent from waste eggshells
 OBJECTIVE

To collect the bio waste egg shell sample

To prepare the dry powder egg shell

To determine the physicochemical properties (ash, pH, moisture and bulk

density)of egg shell powder

To prepare calcium carbonate as bio sorbent from waste egg shells by precipitation

method

To carry out calcination on precipitated calcium carbonate

To characterize the uncalcined and calcined sample by using modern techniques such

as XRD, FT IR, SEM and TG DTA

 To study the adsorption activities on uncalcined and calcined CaCO 3 such

as effects of initial pH, agitation speed, contact time, adsorbent dosage and

operating temperature for removal of heavy metal

 To analyze the concentration of heavy metal in the aqueous solution by

using UV visible spectrophotometer

 To study the adsorption isotherm

 INTRODUCTION
Chicken Eggshells
 Eggshells are waste product. Hens' eggs are used in enormous numbers by food
manufacturers and restaurants and the shells are discarded as waste.

 Many investigations have been conducted to explore useful applications for

eggshells.

 Such research has shown that eggshells may be used as a fertilizer and a feed
additive for livestock and it appears to be able to effectively adsorb certain heavy
metals and organic compounds.

 The porous nature of eggshell makes it an attractive material to employ as an

adsorbent.

 Each eggshell has been estimated to contain7000-17000 pores.

 The principle component of the shell is calcium carbonate which comprise more than
90% of the material.

 The eggshell membrane, which is located between the egg white and the shell is
composed of a network of fibrous proteins that has a large surface area.

 Eggshell membrane has found applications as an adsorbent and as a supporting

medium for immobilization of enzymes (Tsai et al., 2006).
1.2.1 Scientific classification of chicken, Gallus domesticus
Kimgdom : Animalia
Phylum : Chordata
Class : Aves
Order : Galliformes
Family : Phasianidae
Geneus : Gallus
Species : G.domesticus
Scientific Name : Gallus domesticus
English Name : Chicken
Colour : Brown
Max size : 30cm-50cm (12in-20in)
Environment : River, lakes and woodland wetlands
Origin : Italy, India, China, Myanmar and Nepal
Used part :
Classification of chicken eggshells
 The avian eggshell is a porous bio-ceramic that is formed at body temperature in a
cell-free environment.

 Its formation is one of the fastest calcifying processes known in biology.

 The completed chicken eggshell contains about6 g of mineral which is deposited

during its daily production cycle.

 The mineralized shell is about 96% calcium carbonate.

 The remaining components include the organic matrix (2%) as well as magnesium,
phosphorus and a variety of trace elements (Nys et al., 1999).
 The eggshell exhibits extensive intermingling of both its organic and inorganic phases (Chien
et al., 2009).

 This type of organization is also observed in other calcified matrices (bone, cartilage, tooth
enamel, dentin and cementum), where occluded collagenous and noncollagenous elements
are present in intimate contact with mineral.

 The precursors of the eggshell matrix are present in the acellular uterine fluid, from which
they become incorporated into the calcifying shell.

 Eggshell deposition occurs in three stages.

 The entire process lasts almost 17 hours in chicken breeds that are highly selected as layers,
and is the lengthiest phase of egg formation
 The first stage is about 5 hours in duration and corresponds to the initiation of
mineralization.

 The first crystals of calcite are nucleated at the sites of the organic aggregates
present on the surface of outer shell membranes.

 The distribution of these nucleation sites is under genetic control and varies
among species.

 The size of the mammillary cones, the cylindrical diameter of the palisades in the
compact layer of the shell and, ultimately, the strength of the shell is determined
by the spacing of these sites.

 During the first stage of mineralization, crystal growth is radial calcite.

 The nucleation sites become the origins of the mammillary cones.

 As they grow outwards, they gradually come together to form the bases of
the palisade layer, at which point radial crystal growth is inhibited by mutual
exclusion (Nys et al., 1999).

 The second stage is the rapid growth of polycrystalline calcite to form the
palisade layer.

 Formation of the palisades occurs into the available free space, producing
crystals growing perpendicular to the surface of the forming shell, and there is
a linear deposition of about 0.33 g / h of calcium carbonate for about 10
hours.

 The ultrastructure and crystallography of the compact mineral layer can be

partially explained by a single model of competition for crystal growth: growth
of crystals from the nucleation site occurs initially in all directions but, because
of competition for space between adjacent sites of growth , only growing
perpendicular to the egg surface have space to grow.
 This model explains the appearance of preferred crystal orientation in the outer part of
the eggshell, but is based on the hypothesis that the crystal growth is anisotropic.

 Anisotropy results from inhibition of crystal growth on the faces parallel to the c-axis and
results in an elongation of the calcite crystal.

 This inhibition is thought to result from some organic components that are present in the
uterine fluid, and then integrated into the outer eggshell (Hernandez- Hernandez et al.,
2008).

 Another example of this phenomenon, which controls mineral texture, is the preferential
localization of osteopontin on specific calcite crystal faces in the palisades (Chien et al.,
2009).

 The last stage corresponds to termination of calcification and lasts about 1.5 hours (Nys et
al., 1999).
 The arrest of mineralization occurs in a uterine fluid that remains supersaturated in
calcium and bicarbonate ions.

 Proteins are probably implicated since CaCO3 precipitation is inhibited in vitro by high-
molecular weight components of the terminal phase uterine fluid (Gautron et al., 2001).

 Phosphorus is detected in the superficial layers of the chicken eggshell (Cusack et al.,
2003), and spherical aggregates of fine needlelike hydroxyapatite crystals are found in
the outermost calcified layer of the chicken eggshell (Dennis et al., 1996).

 Since phosphate anions can inhibit calcium carbonate precipitation (Lin et al., 2005), the
relative contribution of inorganic phosphate and phosphor proteins to termination of
calcification remains to be determined.

 Ovocalyxin-32, a major phosphor protein of the eggshell matrix, is concentrated in the

outer eggshell and cuticle and therefore is a potential candidate as a proteinaceous
crystal growth inhibitor (Gautron et al., 2001).
 PRECIPITATED CALCIUM CARBONATE

 PCC stands for Precipitated Calcium Carbonate –also known as purified , refined or synthetic
calcium carbonate.

 It has the same chemical formula as other types of calcium carbonate ,such as limestome,
marble and chalk ;CaCO3.

 The calcium, carbon and oxygen atoms can arrange themselves in three different ways,to
form three different calcium carbonate material.

 The most common arrangement for both precipitated and ground calcium carbonates is the
hexagonal form known as calcite.

 A number of different calcite crystal forms are possible:scalenohedral,rhombohedtal and

prismatic.

 Less common is aragonite,which has a discrete or clusture.

 Rare and generally unstable is the vaterite calcium carbonate mineral.

 Precipitated calcium carbonate is the most complex product that we produce: special
controls and a mastery of the parameters involved in the precipitation reaction and later
treatments are required.

 In this way we select with great precision the product features of our Precipitated
Calcium Carbonate(PCC) Calprec desired for every application.

 When comparing to standard Ground Calcium Carbonate (GCC),Precipitated Calcium

Carbonate (PCC) can be distinguished for:

• The possibility to achieve smaller and more uniform particle

sizes
• More stable and narrower granulometric distribution
• Lower abrasivity
• Higher specific surface
• Highest chemical purity.
 In addition the great advantage of Precipitated Calcium Carbonate (PCC) related to GCC
is the possibility of having different crystalline forms which yield to predetermined
physical properties.

 For example,the opacity and whiteness of Precipitated Calcium Carbonate (PCC) is

important in some applications,while in some other cases the rheological behavior is the
fundamental characteristic , consequently the different Precipitated Calcium Carbonate
(PCC) grades are very distinct and the peculliarities of their use in each sector must be
discussed in close collaboration with the customers.
 Benefits of calcium carbonate
 Calcium carbonate is a dietary supplement used when the amount of calcium taken in the
diet is not enough.

 Calcium is need by the body for healthy bones , muscles , nervous system ,and heart.

 Calcium carbonate also is used as an antacid to relieve hearburn,acid indigestion,and

upset stomach.
 Toxicity of calcium carbonate

 Calcium carbonate dust is a physical irritant to eyes , nose , mucous member and skin.

 Contact of calcium carbonate dust with eyes cause redness , pain and inflammation of
the eyelids while contact to skin causes local irritation .

 Exposure of large amount of dust causes coughing , sneezing and nasal irritation.

 Calcium carbonate is an ingredient in most antacids and taken as a dietary supplement .

 Overdose of calcium carbonate may lead to abdominal pain ,bone pain ,coma , confusion
constipation ,depression , diarrhea , headache , irregular heartbeat , muscle twitching ,
nausea and vomiting.
 Application of calcium carbonate

 Calcium carbonate is the common natural form of chalk ,limestone ,and marble ,produced
by the sedimentation of the shell of small fossilized snails ,shellfish and coral over million of
years.

 Precipitated calcium carbonate is used in paper industry.

 It enhances optical properties and print characteristics of paper products, improves paper
machine productivity,and can reduce papermaking costs through the replacement of more
expensive pulp fiber and optical brightening agents.

 Calcium carbonate is the most widely used mineral in the paper ,plastics ,paints and coatings
industries both as a filler and due to its special white colour –as a coating pigment.
 Heavy Metal

 The presence of inorganic pollutants such as heavy metals in aquatic system is of

considerable concern because of their non-biodegradability , mobility , toxicity.

 Heavy metals can accumulate in the human body over time,causing serious health effects.

 Accordingly,the removal of heavy metals from wastewater has received a great deal of
attention in recent years.

 Heavy metals pollution is an environmental problem of worldwide concern.

 The heavy metals are among the most common pollutants found in industrial effluents.
 Cupric Nitrate-Toxicity

 Cupric nitrate can affect you when inhaled.

 Contact can irritate and burn the skin and eyes.

 Inhaling Cupric nitrate can irritate the nose and throat.

 Inhaling cupric nitrate can cause a sore and/or a hole in the bone (septum)dividing the inner
nose.

 Cupric nitrate can cause headache , nausea , vomiting , dirrhea , and abdominal pain.

 Contact may cause a skin allergy.

 Exposure may affect the liver and kidneys.

 Cupric Nitrate is not combustible, but it is a STRONG OXIDIZER that enhances the combustion
of other substances.
 MATERIALS AND METHODS

Preparation of precipitated calcium carbonate

Dissolved in 1000ml of 1M HCL Completely dissolved and adjusted pH of

 100g ESP
solution and used 0.3M NaOH

Filtered using Buchner

filter
continously stirring slowly dropped 1M sodium carbonate
at a constant rate using Filtrate solution
a teflon –coated
magnetic

completed precipitation
and filtered
rinsed several times with D/W dried in an oven at 60.C for
Filtrate solution to remove residual chemical 24hrs and stored in vaccum PESP
desiccator at R/T
Calcination calcium carbonate
Kept for 2hrs calcination in a
 PESP muffle furnace at 650.C Nano sized CaCO3

stored in small air tight bottles

to avoid adsorption of any moisture

calcium carbonate particle

Copper(II)nitrate

dissolved in 250ml of diluted to 1L in Cu(NO3)2

 3.7980g of copper(II)nitrate
deionized water volumetric flask with solution
deionized water
Determination of pH

Placed in a beaker Added 200ml of Placed at R/T Shaken gently

 1g ESP distilled water and filtered

pH solution of Measured by cooled at R/T

fitrate
eggshell using pH meter
Determination of moisture content
Determination by Put in a crucible with a
 Moisture content 1g sample Cover and dried 30.C in
oven method
Oven for 30mins
Cooled in
desiccator

Obtained constant Repeated cooling and

weighing weighing crucible

represented

Moisture percent
Determination of bulk density

 A clean dry 10ml measuring Weighed and filled with Placed in a tapping box
cylinder Sample to 10ml mark
reweighed Tapped gently
Until there is no
More reduction
In volume

recorded

calculated
Bulk density Minium volume
 RESULT AND DISCUSSION

(i) (ii) (iii)

Figure (i) chicken eggshell

(ii) precipitated calcium carbonate
(iii) calcined calcium carbonate
Table-1 Physicochemical Properties of RCESP

Sample Moisture (%) Bulk Density (g cm-3) pH

ESP 0.38 1.48 8.07

ESP- eggshell powder

Table-2 Effect of Concentration of Copper II nitrate on the Removal of
Heavy Metals from Wastewater

Dose of PCC = 0.5g Agitation Speed = 200rpm, pH of wastewater = 5.6

Contact Time = 60min, Volume of Sample = 50ml

Sr.No Concentration of Initial Final Removal% Absorption Capacity

Copper II Nitrate Concentration Concentration
(ppm)
1. 100 3.068 0.484
2. 200 3.214 0.681
3. 300 3.311 0.817
4. 400 3.415 0.932
5. 500 3.800 1.048

Removal % = × 100
 Table-3 Effect of pH of Sample Wastewater on the Removal of Heavy
Metals from Wastewater

Concentration of Copper II Nitrate = 200ppm, Dose of PCC = 0.25g

Contact Time = 60min, Volume of Sample = 50ml

Sr.No pH of Sample Initial Final Removal% Absorption Capacity

(g) Concentration Concentration
1. 3 0.982 0.671
2. 4 1.557 0.742
3. 5 2.742 0.665
4. 6 3.612 0.613
5. 7 2.260 0.733

Removal % = × 100
 Table-4 Effect of pH of Sample Wastewater on the Removal of Heavy
Metals from Wastewater

Concentration of Copper II Nitrate = 200ppm, Dose of Calcine = 0.25g

Contact Time = 60min, Volume of Sample = 50ml

Sr.No pH of Sample Initial Final Removal% Absorption Capacity

(g) Concentration Concentration
1. 3 0.982 0.396
2. 4 1.557 0.543
3. 5 2.742 0.506
4. 6 3.612 0.393
5. 7 2.260 0.556

Removal % = × 100
Table-5 Effect of Dose of Raw ESP on the Removal of Heavy Metals from
Wastewater

Concentration of Copper II Nitrate = 200ppm, pH of wastewater = 6

Contact Time = 60min, Volume of Sample = 50ml, Initial Concentration = 3.612

Sr.No Dose (g) Final Removal% Absorption Capacity

Concentration
1. 0.2 0.561
2. 0.4 0.381
3. 0.6 0.385
4. 0.8 0.384
5. 1.0 0.394

Removal % = × 100
 Table-6 Effect of Dose of PCC on the Removal of Heavy Metals from
Wastewater

Concentration of Copper II Nitrate = 200ppm, pH of wastewater = 6

Contact Time = 60min, Volume of Sample = 50ml, Initial Concentration = 3.612

Sr.No Dose (g) Final Removal% Absorption Capacity

Concentration
1. 0.2 0.703
2. 0.4 0.576
3. 0.6 0.636
4. 0.8 0.789
5. 1.0 0.889

Removal % = × 100
Table-7 Effect of Dose of Calcine on the Removal of Heavy Metals from
Wastewater

Concentration of Copper II Nitrate = 200ppm, pH of wastewater = 6

Contact Time = 60min, Volume of Sample = 50ml , Initial Concentration = 3. 612

Sr.No Dose (g) Final Removal% Absorption Capacity

Concentration
1. 0.2 0.410
2. 0.4 0.359
3. 0.6 0.368
4. 0.8 0.371
5. 1.0 0.376

Removal % = × 100
 Table-8 Effect of Contact Time of Raw ESP on the Removal of Heavy
Metals from Wastewater

Concentration of Copper II Nitrate = 200ppm, pH of wastewater = 6,

Dosage of Raw ESP = 0.05, Volume of Sample = 50ml , Initial Concentration = 3. 612

Sr.No Contact Time(mins) Final Removal% Absorption Capacity

Concentration
1. 10 0.756
2. 30 0.739
3. 50 0.715
4. 70 0.656
5. 90 0.737

Removal % = × 100
Table-9 Effect of Contact Time of PCC on the Removal of Heavy Metals
from Wastewater

Concentration of Copper II Nitrate = 200ppm, pH of wastewater = 6,

Dosage of Raw ESP = 0.05, Volume of Sample = 50ml, Initial Concentration =3.612

Sr.No Contact Time(mins) Final Removal% Absorption Capacity

Concentration
1. 10
2. 30
3. 50
4. 70
5. 90

Removal % = × 100
 Table-10 Effect of Contact Time of Calcine on the Removal of Heavy
Metals from Wastewater

Concentration of Copper II Nitrate = 200ppm, pH of wastewater = 6,

Dosage of Raw ESP = 0.05, Volume of Sample = 50ml , Initial Concentration =3.612

Sr.No Contact Time(mins) Final Removal% Absorption Capacity

Concentration
1. 10 0.459
2. 30 0.424
3. 50 0.389
4. 70 0.438
5. 90 0.445

Removal % = × 100
 UNIVERSITY OF YANGON
DEPARTMENT OF CHEMISTRY
100

95

90

1796.5cm-1
85

1642.3cm-1

80
%T

712.19cm-1
75

70

65
873.03cm-1

60
1407.07cm-1

55
4000 3500 3000 2500 2000 1500 1000 500 400
cm-1
Name Description
Sample I Chit Su San Maung

(a) FT IR spectrum of Eggshell powder (ESP)

 100

95

90

85

80
3641.39cm-1
%T

75 712.83cm-1
873.48cm-1

1411.70cm-1
545.95cm-1
70

65

60

55
4000 3500 3000 2500 2000 1500 1000 500 400
cm-1
Name Cursor Description
Sample II_001 0 %T Chit Su San Maung

(b) FT IR spectrum of precipitated calcium carbonate

 100

95

90

85

80
3641.88cm-1
%T

75
1440.20cm-1
874.65cm-1 712.62cm-1

70

65

60

55
4000 3500 3000 2500 2000 1500 1000 500 400
cm-1
Name Description
Sample III Chit Su San Maung

(b) FT IR spectrum of calcined calcium carbonate

 100

95

90

85

80
%T

75

70

65 870.47cm-1 711.66cm-1
1389.11cm-1

60

55
4000 3500 3000 2500 2000 1500 1000 500 400
cm-1
Name Description
Sample IV_001 Chit Su San Maung

(c) FT IR spectrum of commercial calcium carbonate

Table . FT IR Band Assignments of Precipitated calcium carbonate ,Calcined calcium
carbonate and commercial calcium carbonate

Observed wave number(cm-1) *Literature

wavenumber(cm-1) Band assignment
A B C

3641.39 3641.88 - 3700-3200 Stretching mode of O-H

Band
1411.70 1440.20 1389.11 1395-1440 Bending mode of O-H band

873.48 874.65 870.47 900-650 CO32- ion

712.83 712.62 711.66 730-665 Bending mode of C=C band

545.95 - - 600-500 Stretching mode of C-I

band
 Fourier Transforms Infrared (FT IR) Analysis

 The FT IR spectra of precipitated calcium carbonate(PCC), calcined calcium carbonate and

commercial calcium carbonate are shown in Figure (a) ,(b) and (c) and band assignments are
presented in Table.
 Stretching mode of O-H bands can be found in both samples(a and b) at 3641.39 cm -1 and
3641.88 cm-1 .
 Bending mode of O-H can be observed in the samples(a ,b and c) at 1411.70cm -1,1440.20cm-
1
and 1389.11cm-1. CO32- ion asymmetric stretching can be observed in the samples(a ,b and
c) at 873.48cm-1, 874.65cm-1, 875.71 cm-1, 870.47 cm-1.
 Bending mode of C=C can be found in the samples(a , b and c) at 712.83cm -1 ,
712.62cm-1
and 870.47cm-1 .
 Stretching mode of C-I can be observed in the sample (a) at 545.95 cm -1 .
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