SUMMER TRAINING REPORT

ON

Methyl BlueTremovalTbyTwasteTeggshellTpowder
usingTAdsorptionTprocess
Report submitted in fulfilment for the requirement of the degree
of
B. Tech in Chemical Engineering

BY
Dipesh Joon
Registration No. 02516101419

UNDER THE SUPERVISION

Prof . Deepak Garg

UNIVERSITY SCHOOL OF CHEMICAL TECHNOLOGY GURU

GOBIND SINGH INDRAPRASTHA UNIVERSITY SECTOR 16 C,
DWARKA, NEW DELHI – 110078
2022

i
 CERTIFICATE

This is to certify that the project report entitled “Methyl Blue removal by waste
egg shell powder using adsorption process” which is submitted to the University
School of Chemical Technology, Guru Gobind Singh Indraprastha University,
Dwarka Sector 16 , New Delhi in fulfilment of the requirement for the award of
degree of Bachelor of Engineering in Chemical Engineering embodies the
original research work carried out by Dipesh Joon, Enrollment No:-
02516101419, under the supervision of Dr Deepak Garg. This work has not
been submitted in part or full for any degree or diploma to this or any other
University.

Dr Deepak Garg
(Supervisor)
University School of Chemical Technology
Guru Gobind Singh Indraprastha University
New Delhi-110078

ii
 DECLARATION

I hereby declare that the project report entitled “Methyl Blue removal by waste
egg shell powder using adsorption process” is an authentic record of the
research work and free from any falsification and plagiarism carried out by me
under the supervision of Dr Deepak Garg, University School of Chemical
Technology, GGS Indraprastha University. I shall be solely responsible for any
such dispute arising out of my research work.
Enrollment No. - 02516101419
Date: 29-09-2022

iii
 ACKNOWLEDGEMENT

I consider it my privilege to express gratitude and respect to all those who

guided and inspired me in the completion of my B.Tech project. The
undertaking of this project inculcated a strong sense of research inside me and I
also came to know about so many new things.

First of all, I would like to acknowledge and extend my heartfelt gratitude to Dr

Deepak Garg, Professor at the Department of Chemical Engineering, Guru
Gobind Singh Indraprastha University for his exemplary guidance and
constructive criticism, during the undertaking of this project entitled,

I am especially thankful to all the faculties and supporting staff of the

University School of chemical Technology for their constant help and for
extending the departmental facilities for my project. I would like to extend my
sincere thanks to all my friends for their unconditional assistance and
encouragement. I would also like to keep in record the moral and emotional
support provided by my parents and family throughout the period.

iv
 ABSTRACT

Dyes are a sort of pollutant that can also be seen with the naked eye. However,
dye disposal in valuable water sources ought to be prevented, and diverse
remedy technologies are getting used to perform this. Adsorption is one of the
maximum not unusual strategies for dye elimination. Due to the growing
demand for green and coffee-cost remedy techniques, as well as the importance
of adsorption, low-fee opportunity adsorbents have emerged.

Untreated eggshell powder was used as an adsorbent in this study to remove a

cationic dye, Methyl Blue from an aqueous solution in batch experiments. MB
removal is favored at pH>6 and untreated eggshell powder shows fast
adsorption process and the time required to achieve equilibrium was 40 min.

Inside the batch machine, the effect of various parameters like touch time, pH,
temperature and initial attention were tested on the dye removal from a liquid
medium.

ACRONYMS AND SYMBOLS

v
AR Analytical reagent
MMABL Magnetically Modified Aloe
USEPA United States Environmental Protection
Agency
Temp Temperature
Time Contact time
Min Minute
M Molecular weight
MB Methyl Blue
C0 Initial metal concentration
Ce Final metal concentration
qe Equilibrium uptake of metal
qm Maximum metal uptake or maximum
adsorption capacity
V Volume
RL Separation factor
qt Amounts of metal adsorbed on bio
adsorbent at time

TABLE OF CONTENTS
Contents Page No.
Title Page ……………………………………………………….i
Certificate ……………………………………………………...ii

vi
Declaration ……………………………………………………...iii
Acknowledgement ……………………………………………….iv
Abstract ……………………………………………………....v
Acronyms and symbols ………………………………………...vi
Table of Contents ……………………………………………….vii
List of Figures ………………………………………………..x
List of Tables ………………………………………………..xi

CHAPTER 1
Introduction 1-5

CHAPTER 2
Review of Literature 6-10

CHAPTER 3
MATERIALS AND METHODS 11-19

CHAPTER 4
RESULTS AND DISCUSSION 20-28

CHAPTER 5
Conclusion 29

vii
 CHAPTER 1

Introduction
Methylthioninium chloride, commonly called methylene blue, is a salt used as a
dye and as a medication. Methylene blue is a thiazine dye. As a medication, it is
mainly used to treat methemoglobinemia by converting the ferric iron in
hemoglobin to ferrous iron. Specifically, it is used to treat methemoglobin
levels that are greater than 30% or in which there are symptoms despite oxygen
therapy. It has previously been used for cyanide poisoning and urinary tract
infections, but this use is no longer recommended.

Methylene blue is typically given by injection into a vein. Common side effects
include headache, vomiting, confusion, shortness of breath, and high blood
pressure. Other side effects include serotonin syndrome, red blood cell
breakdown, and allergic reactions. Use often turns the urine, sweat, and stool
blue to green in color. While use during pregnancy may harm the baby, not
using it in methemoglovbinemia is likely more dangerous.

Methylene blue was first prepared in 1876, by Heinrich Caro. It is on the World
Health Organization's List of Essential Medicines.

viii
Methyl Blue (MB) with molecular formula C 16 H18ClN3S

Chemical C16H18ClN3S
formula
Molecular 319.9 g/mol
weight
Λmax 670nm

This compound is prepared by oxidation of dimethyl-4-phenylenediamine in the

presence of sodium thiosulfate.

Methylene blue has been described as "the first fully synthetic drug used in
medicine." Methylene blue was first prepared in 1876 by German chemist
Heinrich Caro.

Its use in the treatment of malaria was pioneered by Paul Guttmann and Paul
Ehrlich in 1891. During this period before the first World War, researchers like
Ehrlich believed that drugs and dyes worked in the same way, by preferentially
staining pathogens and possibly harming them. Changing the cell membrane of
pathogens is in fact how various drugs work, so the theory was partially correct
although far from complete. Methylene blue continued to be used in the second
World War, where it was not well liked by soldiers, who observed, "Even at the

ix
loo, we see, we pee, navy blue." Antimalarial use of the drug has recently been
revived. It was discovered to be an antidote to carbon monoxide poisoning and
cyanide poisoning in 1933 by Matilda Brooks.

The blue urine was used to monitor psychiatric patients' compliance with
medication regimes. This led to interest - from the 1890s to the present day - in
the drug's antidepressant and other psychotropic effects. It became the lead
compound in research leading to the discovery of chlorpromazine.

Methylene blue is used in endoscopic polypectomy as an adjunct to saline or

epinephrine, and is used for injection into the sub mucosa around the polyp to
be removed. This allows the sub mucosal tissue plane to be identified after the
polyp is removed, which is useful in determining if more tissue needs to be
removed, or if there has been a high risk for perforation. Methylene blue is also
used as a dye in chromoendoscopy, and is sprayed onto the mucosa of the
gastrointestinal tract in order to identify dysplasia, or pre-cancerous lesions.
Intravenously injected methylene blue is readily released into the urine and thus
can be used to test the urinary tract for leaks or fistulas.

In surgeries such as sentinel lymph node dissections, methylene blue can be

used to visually trace the lymphatic drainage of tested tissues. Similarly,
methylene blue is added to bone cement in orthopedic operations to provide
easy discrimination between native bone and cement. Additionally, methylene
blue accelerates the hardening of bone cement, increasing the speed at which
bone cement can be effectively applied. Methylene blue is used as an aid to
visualisation/orientation in a number of medical devices, including a Surgical
sealant film, TissuePatch. In fistulas and pilonidal sinuses it is used to identify

x
the tract for complete excision. It can also be used during gastrointestinal
surgeries (such as bowel resection or gastric bypass) to test for leaks.

It is sometimes used in cytopathology, in mixtures including Wright-Giemsa

and Diff-Quik. It confers a blue color to both nuclei and cytoplasm, and makes
the nuclei more visibleWhen methylene blue is "polychromed" (oxidized in
solution or "ripened" by fungal metabolism, as originally noted in the thesis of
Dr. D. L. Romanowsky in the 1890s), it gets serially demethylated and forms all
the tri-, di-, mono- and non-methyl intermediates, which are Azure B, Azure A,
Azure C, and thionine, respectively.This is the basis of the basophilic part of the
spectrum of Romanowski-Giemsa effect. If only synthetic Azure B and Eosin Y
is used, it may serve as a standardized Giemsa stain; but, without methylene
blue, the normal neutrophilic granules tend to overstain and look like toxic
granules. On the other hand, if methylene blue is used it might help to give the
normal look of neutrophil granules and may also enhance the staining of
nucleoli and polychromatophilic RBCs (reticulocytes).

A traditional application of methylene blue is the intravital or supravital staining

of nerve fibers, an effect first described by Paul Ehrlich in 1887. A dilute
solution of the dye is either injected into tissue or applied to small freshly
removed pieces. The selective blue coloration develops with exposure to air
(oxygen) and can be fixed by immersion of the stained specimen in an aqueous
solution of ammonium molybdate. Vital methylene blue was formerly much
used for examining the innervation of muscle, skin and internal organs.The
mechanism of selective dye uptake is incompletely understood; vital staining of
nerve fibers in skin is prevented by ouabain, a drug that inhibits the Na/K-
ATPase of cell membranes.

xi
 CHAPTER 2

Literature review

Adsorption principle
Adsorption is the buildup of a fuel or liquid solute at the floor of a solid or
liquid, ensuing in the formation of a molecular or atomic film. Adsorption is the
adhesion of atoms, ions, biomolecules, or molecules of gasoline, liquid, or
dissolved solids to a surface, and this manner consequences inside the formation
of a movie of the adsorbate (the molecules or atoms being collected) on the
floor of the adsorbent. It is a floor phenomenon due to surface power. Because
the atoms on the adsorbent's floor aren't completely surrounded with the aid of
different atoms, they could appeal to adsorbates. The exact nature of the
bonding depends on the species involved, however the adsorption technique
may be widely categorised as follows:

Adsorption wherein the adsorbate adheres to the floor via Van der partitions
(weak intermolecular) interactions is referred to as physisorption.

xii
Chemisorption is a sort of adsorption wherein a molecule adheres to a surface
by using forming a chemical bond. Adsorption takes place frequently at the
pores' walls or at specific sites within the particle. Because pores are typically
small, the internal floor region is bigger than the outside surface vicinity. Due to
variations in molecular weight, form, or polarity, some molecules are held
greater strongly at the floor than others. In many cases, the adsorbate is held
strongly enough inside the fluid to permit whole removal of that aspect.

The plausible mechanisms of MB adsorption had been additionally proposed.

Evaluations on a few current works on dies

(Deepak et al 2020)

A low-cost activated carbon (PnsAC) was prepared from agricultural waste,

peanut shells, using H₃PO₄ accompanied by using pyrolysis at 650 °C in a
nitrogen surroundings, and became used as a precursor to casting off Acid
Yellow 36 (AY-36) dye from wastewater. The results of pH (2-11), adsorbent
dose (2-6gL1), and initial AY-36 concentration (one hundred-250 mg L1) had
been studied in batch experiments. The isotherms of equilibrium adsorption,
kinetics, and thermodynamics had been studied. Langmuir, Freundlich, Redlich
- Peterson, Sip, and Toth isotherm models have been used to examine the
experimental data. The dye adsorptive elimination kinetics were investigated the
use of pseudo-first-order, pseudo-2d-order, and intra-particle diffusion models.
A have a look at of equilibrium revealed that the Freundlich isotherm version
first-rate defined the experimental statistics. The kinetics of dye adsorption had
been determined to be pseudo-2nd-order, with a correlation coefficient of 0.999.
The adsorption manner was endothermic and spontaneous, consistent with a
thermodynamic observe. Usual, the results confirmed that chemisorption in all
likelihood ruled AY-36 adsorption on PnsAC.

xiii
(Ahmad Alhujaily1,2 et al 2020)

The removal of risky dyes has emerged as a major economic and fitness
problem in wastewater treatment. This observe discovered that efficient, low-
fee, and environmentally friendly spent mushroom waste (SMW) had a
excessive potential for removing anionic dyes including Direct purple 5B
(DR5B), Direct Black 22 (DB22), Direct Black seventy one (DB71), and
Reactive Black 5 (RB5) (RB5). Adsorption mechanisms may be controlled via
each physical and chemical adsorption. SEM analysis revealed that the surface
of SMW is porous and abnormal. The pseudo-2d-order kinetic version
described the kinetic examine properly. The Langmuir version, alternatively, is
the high-quality illustration of the equilibrium isotherm. SMW had most
adsorption capacities of 18 mg g1 for DR5B, 15.Forty six mg g1 for DB22,
20.19 mg g1 for DB71, and 14.Sixty two mg g1 for RB5. The thermodynamic
analysis of G°, H°, and S° validated that the adsorption of the tested dyes onto
SMW become endothermic and spontaneous. Primarily based on these findings,
SMW has a excessive ability as an adsorbent for the removal of dyes from
wastewater.

( Mayank Singh et al 2022)

In this look at, 4 biochars have been evaluated for removal of Remazol terrific
Blue R from the aqueous device: sugarcane bagasse (SBB), coconut shell
(CNB), paddy straw (PDB), and distilled waste of lemongrass (LGB). The
RBBR adsorption capacities of biochar were ninety seven-79% for SBB, 99.9-
ninety nine.47% for CNB, sixty six.1-48% for PDB, and seventy eight-sixty
eight% for LGB, with aromaticity and mineral content dominating. The
chemisorption of RBBR on biochar surfaces has been described the usage of

xiv
Langmuir and Freundlich isotherms, in addition to pseudo-second-order kinetic
fashions. The thermodynamic facts advised that adsorption become endothermic
and spontaneous. Those biochars had been extremely reusable (up to 4 cycles
with 50-sixty one% regeneration). There was no phytotoxicity inside the
purified water or biochar dye sludge. The outcomes of this have a look at may
offer help for the future use of biochars for anionic dye removal from water and
the utilisation of generated sludge for 0 waste-producing technology.

( Rahmiana Zein et al 2020)

In this look at, bovine serum albumin-changed rice husk silica become
organized and examined as an adsorbent for the removal of the metanil yellow
dye. Purification of silica from rice husk become used to attain the silica. To
gain silica, rice husk turned into heated at 700 °C for four hours, and the
ensuing rice husk ash changed into mixed with NaOH five% and acidified with
HNO3 10%. The amendment become completed by using reacting silica with
bovine serum albumin, yielding amine-functionalized rice husk silica.
Fouriertransform infrared, X-ray fluorescence, scanning electron microscopy,
and Brunauer-Emmett-Teller had been used to characterise rice husk silica and
amine-functionalized rice husk silica. The batch method became used for the
adsorption examine, which checked out pH, adsorbate concentration, contact
time, and temperature. Rice husk silica and amine-functionalized rice husk
silica had maximum adsorption capacities of 25.975 and 82.345 mg/g,
respectively. The Langmuir isotherm model was used to describe the adsorption
of metanil yellow dye onto rice husk silica. Furthermore, metanil yellow dye
adsorption onto amine-functionalized rice husk silica was fitted to both the
Langmuir and Freundlich isotherm fashions (R2 = zero.Ninety nine). The
pseudo-2nd-order (R2 = 0.Ninety nine) kinetic facts properly described the
chemisorption process. The thermodynamic equation found out that the

xv
adsorption of metanil yellow dye onto rice husk silica became endothermic,
while the adsorption onto amine-functionalized rice husk silica was
endothermic.

(Megat Ahmad Kamal Megat Hanafiah et al 2018)

Under batch experiments, untreated Aloe vera rind (URAV) changed into used
as an adsorbent to cast off a cationic dye, methylene blue (MB), from aqueous
solutions. PH, URAV dosage, contact time, and MB concentration had been all
numerous to determine the efficacy of MB elimination. At pH > 3, MB was
much more likely to be removed, and URAV exhibited a rapid adsorption
procedure, with an equilibrium time of half-hour. At room temperature, the
maximum adsorption ability decided by way of the non-linear expression of the
Langmuir isotherm model become 356 mg/g (three hundred ok). The pseudo-
first-order and pseudo-second-order models have been each appropriate for
predicting the MB adsorption kinetics. Hydrogen bonding, n- and - interactions,
and electrostatic attraction had been many of the viable adsorption mechanisms
detected by way of the Fourier remodel infrared (FTIR) spectrometer and pH
studies. Due to MB's rapid elimination and excessive adsorption ability, URAV
may be used as an adsorbent for MB removal from wastewater.

xvi
 Chapter 3 Materials and Methods

Glassware and apparatus used

All glassware (conical flasks, measuring cylinders, beakers, pipettes and many
others.) were manufactured by means of
Borosil / Rankem. All of the gadgets and apparatus used within the experiment
are listed beneath:

INSTRUMENT MANUFACTURER/ FUNCTION

TYPE

Electronic weight Aczel To measure weight

balance
Oven Dewsil To dry the samples/ to
maintain the temperature

Shaker Dewsil To shake the samples in

dye solutions

xvii
Spectrophotometer (UV/ Jasco ( V 530) Absorbance
Vis)

pH meter Eutech Instruments Measuring the pH of the

solution

PreparationTofTadsorbent
Rawtchickentshellstaretcollectedtattfasttfoodtrestaurantstandtgrocerytstorestintn
ewtdelhi.tEggtshellstaretwashedtseveralttimestintcleantwatertandtfinallytdriedti
ntatvacuumtoventatt100 degrees Celsius
fortthreethours.tThetmembranetistseparatedtbythand-
driedteggtshells.tFlourtwastobtainedtbytgrindingtittintatflourtmillttotobtaintpow
deredteggtshells.T

PreparationTofTAdsorbateTsolution:T
ThetMBtwastpurchasedtfortsaletintDelhi,tIndia, Acidictacidtay-
36twithtthetchemicaltformula C16H18ClN3Standtatmoleculartweighttoft319.9
gmol-
1thavetbeentusedtwithouttpurificationttotpreparetatsynthetictadsorbatetsolutiont
withtvarioustconcentrationstrangingtfromt10tmgtl-1.tthetconcentrationtoft1tgtl-
1tsolutiontwastpreparedtusingtdoubletdistilledtwater.ttotpreventtphoto-
oxidation,tbrowntglasstbottlestcoveredtwithtaluminumtfoiltaretusedttotstoretthe
tstocktsolution.

xviii
Batch adsorption experiments:
MB was isolated, and a 100 ppm stock solution was made with distilled water.
MB working concentration solutions were prepared through appropriate
dilution. All adsorption experiments were carried out at 303 K room
temperature and 170 rpm stirring rate. At various initial pH values, the effect of
MB on solution pH was investigated (2-10). Each flask was filled with 100 mL
of (100 mg/L) AY solution and 1g of eggshell powder. By adding 0.10 M
H₂SO₄ or 0.10 M NaOH solution, the pH of the AY solution was adjusted.
Thetsolutiontandtphasetweretseparatedtcentrifugationtatt5000trpmtfort5tmin.T

Figure 1Centrifugation at 5000rpm

Thereafter,tthetresidualtdyetintthetsupernatanttwastmeasuredtwithtatuv-
spectrophotometer,twavelengthtofthightabsorptiontMB (λtmaxt=t625 nm).

xix
 calibration curve
0.8
0.7
f(x) = 0.143018181818182 x
0.6 R² = 0.996410221642411
0.5
Absorbance

0.4
0.3
0.2
0.1
0
0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5
Concentration

Effect of Contact Time

In each conical flask, 100 ml of the working solution was placed. All of the
flasks were placed in the shaker at 170 rpm and 30oC for a predetermined time

xx
period ranging from 1 to 60 minutes. The flasks were then removed from the
shaker, the solution was separated from the adsorbents by centrifuge at 5000
rpm for 5 minutes, and the absorbance of the solutions was measured. A graph
of % removal vs. contact time was created.
Effect of pH
From the stock solution, nine working solutions of 100mg/l concentration were
prepared. They were kept at various pH levels ranging from 2 to 8 by adding 0.1
M HCl or NaOH. Each one received 0.5 gram of the adsorbent and was shaken
for 30 minutes at room temperature and 170 rpm in a shaker. After 30 minutes,
the solutions were removed and centrifuged at 5000 rpm before being compared
in a UV spectrophotometer.

CHAPTER 4
Result and Discussion
Effect of pH on AY adsorption:
The pH of dye solution is considered an essential factor in any adsorption
process that affects the adsorption capacity and the adsorption mechanism
of the eggshell. With the objective to investigate the effect of the initial dye
pH in the adsorption of DB78 dye on the eggshell surface, the equilibrium
was studied at different pH values. The effect of pH in the adsorption of
DB78 dye was evaluated in the range of pH from 2 to 8. The pH was
adjusted by adding NaOH or HCl 0.1 M. The pH experiments were
conducted using 1 g eggshell, at a concentration of100ppm, 90 min of
contact time, constant stir (170 rpm), and room temperature (31 °C). The
result indicates that the adsorption capacity and removal efficiency of
eggshell depend on the pH.

xxi
 Effect of ph
88
86
84
82
80
% Removal

78
76
74
72
70
68
1 2 3 4 5 6 7 8 9
Ph

ph %removal absorbance concentration

2 74.82517 3.6 25.17483
4 83.74825 2.324 16.25175
6 85.3007 2.102 14.6993
8 86.92308 1.87 13.07692

Figure 2 Incubrator at 170rpm

xxii
Effect of time:
The rate at which pollutants are removed from aqueous solutions is another
crucial factor in the selection of suitable adsorbents for use in a wastewater
treatment facility. Depending on the concentration of MB, the time required
for AY to achieve adsorption equilibrium has been reported by many studies to
range from a few minutes to a few hours. The effects of different contact times
on the adsorption of MB dye by untreated eggshell powder at the time interval
of 1-60 minutes in 100ml of dye solution , and 100 mg/L of conc at pH 2 were
investigated ,the results are shown below.

Effect of Time
12
10
concemntration

8
6
4
2
0
0 10 20 30 40 50 60 70
time

time conc. absorbance

60 8.713287 1.246
45 8.937063 1.278
30 8.818182 1.261
25 8.783217 1.256
15 9 1.287
10 9.083916 1.299

5 10.28671 1.471
1 10.79021 1.543

xxiii
Figure 3 Different Samples of MB

xxiv
 Chapter 5 Conclusion and future scope

Conclusion
The egg shell membrane powder is an effective material that could be used as
low cost adsorbent to remove anion and cationic dyes (like methylene blue)
from water and wastewater at low concentration and short time. The dye
concentration, the pH of dye solution, the temperature and the particle size of
adsorbent were found as the more effective factors affecting the adsorption
process. There is a need for new researches to develop the combination between
egg shell membrane and other industrial dyes for more applications.
Future Scope
Thustittcantbetconcludedtthatteggshells,twhichtaretwastetmaterialstandtaretinta
abundancetintthetcountry,tcantbetusedtfortthetremovaltoftMBtdyetfromtwaste
water further ,there is scope of more study and research which can enhance the
percentage removal of dye increased by different treatment of adsorbent.t

xxv
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xxix