Bahir Dar University

College of Science− Postgraduate Program

Department f Chemistry

MScThesis Proposal on
Simultaneous Electrochemical Determination of
Cadmium, Lead and Mercury in Bahir Dar
Textile Factory Effluent Using Silver
Nanoparticles Modified Glassy Carbon Electrode

By: Awoke Worku

Advisor: Meareg Amare ( PHD)
November 2018

1
 Bahir Dar,Ethiopia

2
 Approval Sheet

I, the under signed MSc student declare that this thesis proposal which is conducted in

partial fulfillment of the degree of master of Chemistry (Analytical Chemistry) at Bahir Dar
University is my respective work.

Submitted by:

Name of Student: Signature: Date:

1. Awoke Worku ____________ ______________________

Approved by:
2. Name of Advisor: Signature: Date:

__Meareg Amare (PHD) _______________ _____________________

Name of Evaluators: Signature: Date:

1 __________________________ ____________ _______________

2 __________________________ ____________ ________________

3 ___________________________ ______________ ____________________

Name of Department Head: Signature: Date:

________________________ ____________ ________________

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 Approval Sheet.....................................................................................................................................ii
ABSTRACT............................................................................................................................................iv
1 INTRODUCTION...........................................................................................................................1
1.1 Background of the study........................................................................................................1
1.2 Statement of the Problem......................................................................................................4
1.2.1 Health and Environmental Impact of Cadmium 4
1.2.2 Health and Environmental Impact of Lead 5
1.2.3 Health and Environmental Impact of Mercury 5
1.3 Objective of the Study............................................................................................................6
1.3.1 General Objective 6
1.3.2 Specific Objective 6
1.4 Significance of the Study........................................................................................................7
2 LITERATURE REVIEW...................................................................................................................7
3 MATERIALS AND METHODS........................................................................................................8
3.1 Method..................................................................................................................................8
3.1.1 Chemicals and apparatuses required 8
3.1.2 Sampling 8
3.1.3 Sample Collection and Preparation 8
3.2 Data Collection.......................................................................................................................9
4 TIME FRAME...............................................................................................................................9
4.1 Activities and Frame of the Research Project.........................................................................9
4.2 Budget breakdown of the study...........................................................................................10
4.2.1 Stationery Expenditure 10
4.2.2 Cost of Reagents 10
4.2.3 Sample Analysis 11
4.3 Budget Summary..................................................................................................................11

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ABSTRACT

The principal objective of this research is synthesizing silver nanoparticles for the purpose
of augmenting the sensitivity and selectivity of glassy carbon electrode for determining and
detecting analytes at extremely low level. Heavy toxic metal cations namely: cadmium(II)
ion, lead(II) ion and mercury(II) ion that are being discharged from industries to the
nearby receiving water bodies which trigger severe health and environmental turmoil will
be determined sing AgNPs modified glassy carbon electrode. Electrochemical method in
general and cyclic voltammetric method in particular in which electrochemical cell
consisting of three electrode system will be applied to determine the toxic heavy metal
cations.Friendliness with the environment, swift response to the concentration of
analyte,specificity to a particular oxidation state, portability of the instruments, low
detection limit and low cost vis-à-vis other methods are the fundamental factors that made
electrochemical methods preferable.
Simultaneous Electrochemical Determination of Cadmium,Lead and Mercury in Bahir
Dar Textile Factory Effluent Using Silver Nanoparticles Modified Glassy Carbon
Electrode

1 INTRODUCTION

1.1 Background of the study

Environmental Pollution is unwanted change in physical, chemical and biological

characteristics of air, water and soil which is harmful for both animals and plants.
Pollution can take the form of chemical substances or energy, such as noise, heat or light.
Pollutants, the elements of pollution, can be either foreign substances/energies or naturally
occurring contaminants. Heavy metals are usually present in trace amount naturally in
water but many of them are toxic even at very low concentration. Metals such as arsenic,
lead, cadmium, nickel, mercury, chromium, cobalt, zinc and selenium are highly
toxic even in minor quantity. Increasing quantity of heavy metals in our resources is
currently an area of greater concern especially since a large number of industries are
discharging their metal containing effluents in to fresh water without any adequate
treatment. Heavy metals become toxic when they are not metabolized by the body and
accumulate in the soft tissues [1]. Toxic heavy metals identified in the effluent of textile
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factory include As, Cr, Cu, Hg, and Pb that are poisonous for human and aquatic life [2].
In spite of the availability of substitutes for lead and cadmium compounds in paints,
manufacturers continue to produce paints with high levels of these metals. As the
population continues to grow and there is a continued shift from oil-based to water-
based paints, the sales and use of these paints will increase the exposure of humans and
the environment to these metals [3].As per the standard that has been set by EPA in
2003,the maximum permissible limit,MPL of Co,Cr,Pb,Cd,Zn,Fe,As and Hg in textile
effluent before it is discharged to the environment is
1000,1000,500,1000,50001000,250,and 1.0 ppb respectively [20].Electrochemical
methods have been chosen to analyse the toxic heavy metal cations.Despite the
difference in instrumentation, all electrochemical techniques share several common
features [4].Electrochemical processes are commonly used for analytical

measurements. There are a variety of electrochemical methods with different

degrees of utility for quantitative and qualitative analysis. The methods rely on one
of the two different electrochemical phenomena. The first is that many chemical
species have the ability to transfer electrons through oxidation-reduction process.
With appropriate design of an electrochemical system, this transfer of electrons can
be measured as a current. Since we also know that different species have different
oxidation or reduction abilities, electrochemical measurements relying on electron
transfer can often be used for the purpose of species identification. The second
method of using electrochemical processes for measurement purposes relies on the
measurement of a potential. In particular, methods that depend on junction potential
will be given special emphasis. Acquaintance with electrochemical cells is
coercionary to deal with junction potential. Such a device consists of electrodes and
the design of electrodes creates interfaces or junctions (e.g., a metal electrode in
contact with a solution represents a junction). Any junction in an electrochemical
system will have a potential associated with it and in certain cases, the magnitude of
this junction potential can be related to the concentration of a species in solution. For
example, a pH electrode is the best known example of the use of a junction potential

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 for determining the concentration of a species. The key feature of a pH electrode is a
thin glass membrane. When placed into an aqueous solution, a junction potential
occurs at the glass solution interface and the magnitude of this potential is
determined by the concentration of H+ in solution [5].Voltammetric techniques such
as cyclic voltammetric techniques have been reported as flawless techniques in
analyzing trace metals in real samples [6].

Nanoparticles

The term “nano” is derived from the Greek word “nanos” for “dwarf”. This
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etymology, and its placement on the metric scale (1nm = 10 m), shows that tiny
dimensions not visible to naked eye, beyond the normal limits of our observation. The
diameter of hair (100 µm) to the buck-minster fullerene (1 nm) shows the variation of
diameter of a material. Nanoscale materials are defined as materials having at least one
dimension in the 10–100 nm range, in which nanoparticles are the major investigated
subjects. The term “nano-particle” (earlier termed small particles first appeared in the
1980s. The nanoscience deals with the materials having particle size in nanometer
regime and their properties. The interdisciplinary nature with contributions from
chemistry, physics, biology, materials sciences, all the way to engineering and medicine is
one of the most striking features of the nanosciences when compared to many classical
disciplines, and is important for the development of nanotechnological products[7]. The
field of nanoscience is one of the most rapidly expanding areas of research across all
disciplines. This is due largely to a wide range of applications for nanoparticles, such as
electronic devices, sensors and biomedical devices

Classification of Nanoparticles

There are various bases of classifying nanoparticles.Nanoparticles can be classified based on

the following parameters.

 Based on origin
 Based on size
 Based on chemical composition
Based on origin, nanoparticles are classified in to natural and anthropogenic.
Based on size, nanoparticles are classified in to from 1−10 nm,from 10−100 nm and
above 100 nm
Based on composition, nanoparticles are classified in to organic substances,
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 inorganic substances and nanoparticles of living kingdom.

PREPARATION OF NANOPARTICLES

There are two ways by which nanoparticles can be synthesized.

 Bottom up Approach
 Top down Approach
a) Bottom up Approach

In bottom-up approach or ‘self-assembly’ (assembling particles synthesized in

solution) is a process universal in nature, atoms and molecules are assembled from
molecular levels so as to have nanomaterials of desired size and shape by controlled
reaction parameters. Wet chemical synthesis is the most energy efficient ‘bottom up’
technique for the synthesis [7].

b) Top Down Approach

In top down approach, the reverse is the case. The conventional approach involves
chemical or physical attrition from bulk into objects of desired sizes and shapes (e.g.
lithography, mechanical milling, ion implantation, etc.), and is known as the ‘top-down’
approach [7]

1.2 Statement of the Problem

1.2.1 Health and Environmental Impact of Cadmium

Acute and chronic exposure for cadmium gives rise to health complications for both human
beings and animals. As per EPA (1996) cancer guideline, cadmium has been categorized as
potential carcinogen. Cadmium can enter our body by smoking, inhaling air, through food or
drink. About 200,000 sorts of chemicals are being produced every year across the globe and
the toxicity of the majority of these chemicals is not known. The interaction of chemicals
and human beings takes place through air, food and drink. Despite the discharge of large
amount of cadmium to the environment every year, about 25,000 ton a year. About half of
this cadmium is discharged in to rivers through weathering of rocks and the remaining
viii
cadmium is emitted in to air through forest fire and volcano. Industrial sources such as
mining, ,refining of ores and the plating process have great contribution for the emission of
cadmium in to the environment. Cadmium is toxic to the aquatic environment and non
essential metal. [1]Cadmium has various uses such as for the production of cadmium
batteries, anti corrosive coating of metals, pigments and stabilizers for plastics. The
maximum permissible limit of cadmium in drinking water is 5 ppb. Renal damage,
osteoporosis and possibly renal cancer are health complications arising from oral exposure
to cadmium. Chronic exposure even to very low amount of cadmium results in adverse renal
and negative bone effects. Itai-itai disease was the combined effect of renal and bone
effects. In Japanese ‘itai’ means ‘ouch’or painful in English. The agony results from
uncommon changes in bone,i.e.osteomalacia with osteoporosis. In May 1968,Ministry of
Health of Japan described that the cause of itai itai disease was osteomalacia with
simultaneous renal dysfunction from chronic cadmium poisoning and it was under the
influence of the following factors: pregnancy, lactation, hormonal disorders, ageing, calcium
deficiency and others[9]. As per US EPA the maximum permissible level of cadmium in
textile effluent is 1000 ppb[13].

1.2.2 Health and Environmental Impact of Lead

Lead is the most abundant of the heavy metals in the Earth’s crust. It has been u sed
s ince prehistoric t imes, and h as become w idely distributed a nd mobilized in the
environment. At h igh levels of human exposure there is damage to almost all
organs and organ systems, most importantly the central nervous system, kidneys
and blood, culminating in death at excessive levels. Lead poisoning was common
in Roman times because of the use of lead in water pipes a nd earthenware
containers, and in wine storage. Case-control studies o n mental retardation and
h yperactivity in relation to environmental lead exposure showed that children w ho
survived a cute lead intoxication w ere often left with severe deficits in
neurobehavioural function. It was subsequently recognized that long- term sequelae
were not limited to people affected by excessive exposure but also occurred in
children who experienced r elatively low-level exposure. Lead levels in h uman
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 skeletal remains indicate that t he body lead burden of today’s populations is 500–
1000 times g reater t han that of their pre-industrial counterpart. Debate
continues over the nature, magnitude and p ersistence of t he a dverse effects
on h uman health of low-level exposure t o environmental lead. However, the
accumulated epidemiological evidence indicates t hat such exposure i n early
childhood causes a discernible deficit in cognitive development during the
immediately ensuing c hildhood y ears. Neuropsychological manifestations will
be disappeared or d eclined if the ingestion of lead was stopped o r reduced.
R ecent data, however, show that s uch e ffects are largely irreversible [10].

1.2.3 Health and Environmental Impact of Mercury

Sources of exposure are wide and include mercury vapors in ambient air, ingestion via
drinking water, fish, vaccines, occupational exposures, home exposures including fluorescent
light bulbs, thermostats, batteries, red tattoo dye, skin lightening creams, and over the counter
products such as contact lens fluid and neosynephrine, dental amalgams, and more. Amalgam
exposure is estimated to be from 3 to 17 micrograms per day from slow corrosion, chewing,
brushing and grinding. The clinical manifestations are varied and mimic many other
conditions. Central Nervous System (CNS) toxicity includes erethism with symptoms of
shyness, emotional lability, nervousness, insomnia, memory impairment, and inability to
concentrate.

Other CNS symptoms may include encephalopathy, peripheral neuropathy,

Parkinsonian symptoms, tremor, ataxia, impaired hearing, tunnel vision, dysarthria,
headache, fatigue, impaired sexual function, and depression. Renal toxicity includes
proteinuria, renal syn- drome, and acute renal failure. Gastrointestinal symptoms include
nausea, vomiting, diarrhea, and colitis. Dermal toxicity includes allergic dermatitis,
chelitis, gingivitis, stomatitis, and excessive salivation [11].

Some advanced analytical techniques such as AAS are not specific to a particular
oxidation state of the species rather they impart general information. They give total
concentration of the different oxidation states of that heavy metal. For instance,
hexavalent chromium and trivalent chromium cannot be analyzed using AAS separately
rather their total concentration will be determined.Moreover,they are costly. The cost of
analysis per unit sample is very high. On the contrary, electrochemical methods are

x
 specific to a particular oxidation state of a species, their cost of analysis per unit sample is
relatively cheap vis-à-vis spectroscopic technique

1.3 Objective of the Study

The purpose of this research is determination of toxic heavy metals in textile effluent by
electrochemical method simultaneously using modified electrode.

1.3.1 General Objective

To determine toxic heavy metal cations namely:cadmium (II) ion,,lead (II) ion and
mercury (II) ion in the effluent of Bahir Dar Textile Factory

1.3.2 Specific Objective

 To determine the toxicity level of heavy metal containing effluent of Bahir
Dar Textile Factory by comparing it with the standard
 To determine cadmium (II), lead (II) and mercury (II) ions in Bahir Dar
textile effluent at a very low level using silver nanoparticles modified carbon
paste electrode
 To render recommendation as to how these toxic heavy metal cations can be
dwindled and removed totally from textile effluent
 To analyze the efficiency of the synthesized Nanoparticles in boosting the
sensitivity of carbon paste electrode in detecting the heavy metal cations in
Bahir Dar textile factory at extremely low level by characterizing them with
FTIR,SEM,UV and XRD

1.4 Significance of the Study

The importance of this research is to make the society conscious about the
environmental and health problems that result from industrial effluents that are being
discharged from miscellaneous industries to the nearby receiving water bodies. Once
the society knows the health problems resulting from heavy metal cations, they will
safeguard themselves either from acute or chronic exposure to these toxic ions.
Farmers will refrain from harnessing rivers to which untreated or inadequately treated
effluents are being discharged for irrigation purpose.This project will render
recommendation as to how human health and environment can be safeguarded as well.
Once the society discerns the detriment of these industrial effluents, they will refrain

xi
 from using them unless they are treated adequately. This research work can also serve
as a reference material for prospective researchers to bridge the gap which is not
addressed in this research for various constraints. This research is used to determine
the toxicity level of heavy metals in the effluent. The observed and actual results will
be compared.

2 LITERATURE REVIEW

Several methods of determining trace heavy metal cations in industrial effluents have been
reported. One of the methods is electrochemical method which makes use of current,
potential (voltage) and charge to quantify the amount of analyte that is present in a
particular sample. Cyclic voltammetry one of the most convenient technique of
electrochemical methods is being applied extensively for the determination of heavy metal
cations such as lead mercury and chromium in industrial effluents owing to its large
potential range to measure current [12].It has been described in many publications that
heavy metal cations that are found in industrial effluents can give rise to serious health and
environmental problems. As per the research that has been conducted on Hawassa Lake
signifies aquatic life such as fish have been jeopardized due to the textile effluent that enters
the lake [13]. A number of international publications signify the toxicity of heavy metals
which are contained by industrial effluents as they are non degradable and they are not
metabolized by the body [1].

3 MATERIALS AND METHODS

3.1 Method

Green synthesis method will be used to synthesize silver nanoparticles. The extract from
Ocimum Sanctum (holy basil) leaf will be used for reducing silver cation.There are two
reasons to select Ocimum Sanctum leaf as reducing agent.The first reason is it is
eco−friendly.The second reason is it acts as stabilizing agent for the nanoparicle owing to
the presence of bio molecules inside it.UV,FTIR, SEM and XRD instruments will be used
to characterize the synthesized silver nanoparticles. Cyclic voltammetric technique will be
used to determine the toxic heavy metal cations namely: cadmium (II)ion, lead(II) ion and
mercury(II) ion. Three electrode system voltammetric measurements will be conducted.
Three electrodes namely: the counter electrode (often platinum wire), the reference
electrode (Ag/AgCl) electrode, and the working electrode (AgNPs modified glassy carbon

xii
electrode ) will be immersed in to electrochemical cell containing the toxic metal cations
under examination.

3.1.1 Chemicals and apparatuses required

Silver nitrate (AgNO3), ocimum sanctum leaf extract, distilled water,deionized

water,standardCd(NO3)2,standardPb(NO3)2standarHg(NO3)2,conc.HNO3(63%), 30%
H2O2,buffer solution of standard PH

Apparatuses Instruments

Pipettes, micropipettes, measuring cylinders, beakers, volumetric flasks, fume hood, hot
plate, kjeldahl setup, plastic bottles, thermometer, PH meter, conductivity meter, three
electrode system electrochemical cell connected to computer, UV spectrometer, FTIR,SEM

3.1.2 Sampling
Time weighted representative composite sample will be taken from the discharge point of
the textile factory by dividing the sample taking time in to equal intervals. The sampling
time is based on the discharging time of the effluent to the nearby water body. When the
amount of toxic metal cations in the effluent is assumed to be very high and when the
amount of toxic metal cation in in the effluent is assumed to be very low.

3.1.3 Sample Collection and Preparation

Five effluent samples will be collected with polyethylene plastic bottle that is rinsed with
10% HNO3 from the discharge point of the factory at different time intervals at 12 O’clock
in the morning, at three O’clock, at six O’clock, at nine O’clock and at 12 O’clock in the
evening and then the five samples will be mingled together with 500 mL polyethylene
plastic bottle to make time weighted representative composite sample. The most important
water quality parameters namely: PH, temperature, dissolved oxygen (DO), electrical
conductivity (EC) and turbidity will be measured in SITU. The time weighted composite
sample will be digested with nitric and perchloric acid to eliminate interferences and to get
lipid solution and it will be kept in the refrigerator until analysis.

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 3.2 Data Collection

Data about the shape and size of the synthesized nanoparticles will be collected
from SEM. Data about strong absorption of nanoparticles will be collected
from UV spectrometer.Data regarding adsorption of functional groups on the
surface of nanoparticles will be obtained from FTIR spectroscopy.Data about
the concentration of trace heavy metal cations in the the effluent sample will
be collected from three electrode system measurement of electrochemical cell.

4 TIME FRAME

4.1 Activities and Frame of the Research Project

Table one: List of Activities and their Period of Accomplishment

NO Type of Activity Period of Accomplishment

Sep Oct Nov Dec Jan Feb Mar Apr May Jun
1 Selecting Title
2 Proposal Writing
3 Submission of Proposal
4 Defending Proposal
5 Collecting Sample
6 Purchase of Chemicals and
Apparatus
7 Preparing Sample
8 Conducting Experiment and
Analyzing Data
9 Writing Thesis
10 Defending Thesis

4.2 Budget breakdown of the study

4.2.1 Stationery Expenditure

Table Two: Budget allotted to Stationery

Ooo0 Items Description Unit Qty Unit Cost Total Cost Remark
oO (Birr)
(Birr)
Re-
1 Rewritable CD 700 MB Pcs 5 25.00 125.00
2 Photocopy Proposal, Progress Page 300 0.50 150.00

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 R report and thesis copies

Pen
3 pen Lexi Pcs 6 6.00 36.00

4
Printing Journals Page 150 1.00 150.00
5 printing papers Page3 300.00 1.00 300.00
Sub Total 761.00

4.2.2 Cost of Reagents

Table Three: Cost of Chemicals,Reagents and Transportation

NO Item Description Unit Qty Unit Total Cost Remark

Cost (Birr)
(Birr)
1 30% H2O2 Liter 1 620.00 620.00
2 63% HNO3 Liter 1 980.00 980.00
3 Silver Nitrate Crystal G 100 85.00 8500.00
4 Standard Cd(NO3)2 G 100 9.60 960.00
Crystal
5 Standard Pb(NO3)2 g 100 2.40 240.00
Crystal
6 Standard Hg(NO3)2 G 100 10.60 1060.00
Crystal
7 Transportation 1000.00
expenditure from
Bahir Dar to AA and
the other way round
Sub Total 13,360.00

4.2.3 Sample Analysis

Table Four: Cost of Sample Analysis

NO Description Unit Quantity Cost/Sample Total Cost Remark

(Birr) (Birr)
1 FTIR Analysis sample 5 100.00 500.00
2 SEM Analysis sample 5 2000.00 10,000.00
3 UV-Analysis sample 5 40.00 200.00
4 XRD Analysis sample 5 500.00 2500.00
Sub total 13,200.00

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4.3 Budget Summary

Table Five: Budget summary

NO Budget Title Total Cost Remark

(Birr)

1 Cost of Chemicals, Reagents and 13,360.00

Transportation

2 Cost of Sample Analysis 13,200.00

3 Cost of Stationery Materials 761.00

Grand Total 27,321.00

Approved by:

Name Signature: Date:

_Meareg Amare (PHD) _______________ ________________

Name of Internal Examiner: Signature: Date:

1 ____________________ ____________ ___________________

Name of External Examiner: Signature: Date:

2 ____________________________ ____________ ___________________

xvi
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