DETERMINATION OF HEAVY METALS IN (LETTUCE)

BY

MUHAMMAD BUHARI SANI

2019/ND/SLT/238

SUBMITTED TO DEPARTMENT OF SCIENCE LABORATORY AND

TECHNOLOGY THE FEDERAL POLYTECHNIC DAMATURU, YOBE
STATE

MARCH, 2022.

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 APPROVAL PAGE
This Project has been read and approved by undersign person on behalf of the
Department Of Science Laboratory Technology, The Federal Polytechnic,
Damaturu.

MUHAMMAD HUSAIN

SUPERVISOR

DATE-----------------------

SIGN-----------------------

EXTERNAL EXAMINER

DATE……………………..

SIGN……………………..

HOD: DR. IBRAHIM BABALE GASHUA

DATE-----------------------------

SIGN-----------------------------

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 DEDICATION
This project is dedicated to almighty Allah who gave me the opportunity to write
this project and with gratitude to my lovely parent.

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 ACKNOWLEDGMENT
All praise be to almighty God who sustained our live to witness this project end.

Furthermore, praise and salutations be to his Messenger (PBUH).

I will like to first give regards to my parents for their utmost support and
encouragement throughout my life. The effort of my supervisor Mal. Muhammad
Hussaini and Head of department Dr. Ibrahim Babale Gashua will never be in vain,
I will always carry it along in my hearts.

More regards are also given from me to the entire staffs of Science and Laboratory
Technology Department for their selfless and hard work towards our success in our
programme duration.

I also recognized the caring, love and support to my family and friends, I give
regards to you and I said a big thank you. May the almighty support you too.

TABLE OF CONTENTS

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 APPROVAL PAGE......................................................................................................................................ii
DEDICATION............................................................................................................................................iii
ACKNOWLEDGMENT...............................................................................................................................iv
TABLE OF CONTENTS...............................................................................................................................v
ABSTRACT..............................................................................................................................................vii
CHAPTFR ONE..........................................................................................................................................1
1.0 INTRODUCTION...........................................................................................................................1
1.1 HISTORY OF HEAVY METALS........................................................................................................2
1.2 HEAVY METAL RELATIONSHIP TO LIVING ORGANISM.................................................................2
1.3 ` AIM OF THE STUDY....................................................................................................................3
1.4 OBJECTIVES OF THE STUDY..........................................................................................................3
1.5 SIGNIFICANCE OF THE PROJECT WORK........................................................................................3
1.6 STATEMENT OF PROBLEM...........................................................................................................3
1.7 SCOPE AND LIMITATION.............................................................................................................4
CHAPTER TWO.........................................................................................................................................5
2.0 LITERATURE REVIEW....................................................................................................................5
2.1 HAZARDOUS EFFECT OF HEAVY METALS ON HUMAN HEALTH....................................................6
2.2 THE RISK OF CONTAMINANTS ACCUMULATING IN SOIL ENVIRONMENT AND CROPS................8
2.3 CHILDHOOD EXPOSUER TO HEAVY METALS CAN RESULT IN LEARNING DIFFICULTIES, MEMORY
IMPAIRMENT AND MORE........................................................................................................................8
2.5 ACUTE TOXICITY OF COPPER........................................................................................................9
2.6 SYMTOMS OF EXPOSURE TO TOXIC METALS...............................................................................9
CHAPTER THREE.....................................................................................................................................11
3.0 METHODOLOGY.........................................................................................................................11
3.1 MATERIAL..................................................................................................................................11
3.2 REAGENTS..................................................................................................................................11
3.3 SAMPLE COLLECTION.................................................................................................................12
3.4 GLASS WILL BE SAMPLE AND PRE - TREATMENT.............................................................................12
3.5 NITRIC- PERCHLORIC ACID DIGESTION.......................................................................................12
CHAPTER FOUR..........................................................................................................................................13
4.0 HEAVY METAL ANALYSIS............................................................................................................13
CHAPTER FIVE........................................................................................................................................14
REFERANCES..........................................................................................................................................15

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 APPENDIX..............................................................................................................................................17

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 ABSTRACT
Vegetables constitute an important parts of the human diet since they contain
carbohydrate, proteins, vitamins, minerals and fibres required for human health
(yusuf, 2003). Living organism require varying community of heavy metals, iron
(Fe), cobelt (Co), copper (Cu) etc will be required by humans. Excessive levels con
cause damage to the organisms. The aim of this project is to determine the
presence of heavy metals In selected vegetables, the level of heavy metals
contamination and to provide the way out to minimized the risk of those heavy
metals contamination and it is also significant to monitor the heavy metals
contamination in our environment in order to regulate the bioaccumulation
properties which could result in health effect. Two leafy vegetables specie s
(Brassica oleracea linn. Var. capitata). Commonly found in Damaturu farm lands,
were analyzed for copper (Cu) and cadmium (Cd) content by Atomic absorption
spectrophotometer. However the results showed that (he level of (cu) range from
0.05ug/g to 0.07ug/g and 0.025wg/g to 0.05ug/g for cadmium in a„ the vegetables
sample including the soil.

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 CHAPTFR ONE

1.0 INTRODUCTION
Vegetables constitute an important part of the human diet since they contain
carbohydrates, proteins, vitamins, minerals and fibres required for human health
[Yusuf, 2003], They also acts as neutralizing agents for acidic substances formed
during digestion [Thompson and Kelly, 2013].

As human activities increase, especially with the application of modern

technologies, pollution and contamination of the human food chain has become
inevitable. Heavy metals uptake by plants grown in polluted soils has been studied
to considerable extend (Wong, 2014).

Heavy metal exposure is not an entirely modern phenomenon; Historians have

cited the contamination of wine and grape drinks by lead-lined jugs and cooking
pots as a contributing factor in the decline and fall of the Roman Empire and the
mad hatter character in Alice in wonderland will be likely modeled after nineteenth
century hat markers who used frequently become psychotic from mercury toxicity
(Radman and Saiama, 2017). Human exposure to heavy metals has risen
dramatically in the last, 50 years as a result of a exponential increase on the use of
heavy metals in industrial process and products (Wong, 2006).

Heavy metal contamination in vegetables cannot be under estimated as these food

stuffs will be important components of human diet. Heavy metals contamination of
the food items is one of the most important aspects of food quality assurance
(Mashall 2004).

International and national regulations of food quality have lowered the maximum
permissible levels of toxic metals of food items due to an increased awwill beness

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of the risk, these metals pose to food chain contamination (Radwan and Salama,
2006).

1.1 HISTORY OF HEAVY METALS

The toxicity of these metals has also been documented throughout history, Greak
and Roman physician’s diagnosed symptoms of acute lead poisoning long before
toxicology become a science. Today, much more is known about the health effects
of heavy metals (Jwill bemo, 2009).

Exposure to heavy metals has been linked with developmental retardation, various
cancers’ kidney damage, and even death in some instances of exposure to high
level of mercury, gold and lead has also been associated with development of
autoimmunity, in which the immune systems start to attack its own cells, mistaking
them for foreign invaders (Adeniyi, 2000).

Since the industrial revolution, the production of heavy metals such as lead, copper
and zinc has increased exponentially between 1850 and 1990, production of the
three metals increased nearly 10-fold, with emission rising in tandem [Adeniyi,
2006].

1.2 HEAVY METAL RELATIONSHIP TO LIVING ORGANISM

Living organism require varying community of heavy metals, iron, cobalt, copper,
manganese, and zinc will be required by humans. Excessive levels can be
damaging to the organism. Other heavy metals such as mercury, plutonium and
toxic metals that have no known vital or beneficial effect on organisms and their
accumulation over time in the bodies of animals can cause serious illness. Certain
elements that will be normally toxic, for a certain organisms or under certain
conditions, beneficial. Examples include vanadium tungsten, and even cadmium
[Khan, 2008].

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1.3 ` AIM OF THE STUDY
The aim of this study is to determine the presence of heavy metals in some selected
vegetables.

1.4 OBJECTIVES OF THE STUDY

I. To determine the presence of heavy metals in

To determine the levels of heavy metal contaminations in some selected

vegetables.

II. To provide the way out in minimizing the risks of heavy metals
contamination.

1.5 SIGNIFICANCE OF THE PROJECT WORK

The significance of this study work is to assess level of different heavy metals in
selected vegetable samples in order to monitor the heavy metals contamination in
our environment and also to regulate the bioaccumulation properties which could
result in health effects.

1.6 STATEMENT OF PROBLEM

Exposure to potentially toxic metals from dust inhalation or soil ingestion is
usually modeled simply as the concentration of a contaminant measured in the soil
multiplied by the quality of dust inhaled or soil ingested. This is a conservative
approach to estimate dose because the bio-accessibility of heavy metals absorbed
on ingested soil is not 100%.

However, predicting exposure to potentially toxic metal from consumption of food

crops is more complicated because uptake of metal by plants depends on soil
properties and plant physiological factors. This lead to much larger uncertainties
associated with estimating potential doses through food chains compwill bed to the

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uncertainties associated with other exposure pathways such as soil ingestion and
dust inhalation.

1.7 SCOPE AND LIMITATION

There will be several heavy metals that could be found in vegetables. This research
limited only to consider cadmium and copper. This research limited also considers
vegetables found locally in irrigated farms within Damaturu metropolitan. This
research work is however, limited to the apparatus, equipment, reagents and
instruments available in the department of science laboratory technology (SLT).
Federal Polytechnic Damaturu, as well as time of the program.

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 CHAPTER TWO

2.0 LITERATURE REVIEW

Rapid and unorganized industrialization and urbanization have contributed to the
elevated levels of heavy metals in the urban environment of the developing
countries such as china and India (Wang and Killwill be, 2003).

Heavy metals will be non-biodegradable and persistent environmental

contaminants which may be deposited on the surfaces and then adsorbed into the
tissue of the vegetables. Plants take up heavy metal by absorbing them from
deposits on the parts of the plants exposed to the air from polluted contaminated
soils (Khairich, 2004).

Water contamination by heavy metals in some will beas is practically inevitable

due to natural process [weathering of rocks] and anthropogenic activities.
[Industrial], [Agriculture and domestic effluents]. Will bete water from the
industries of mining, electroplating, paint or chemical laboratories often contain
high concentrations of heavy metals including cadm.um (cd),copper (cu) and lead
(Pb). These elements, at concentration exceeding the physiological demand of the
Plants, not only could enter food chain, get biomagnified and pose a potential
threat to human health (Sugiyama, 2005).

Heavy metals contamination of agricultural soil from will bete water irrigation is
of serious concern since it has implications on human health. A study carried by
Mensa (2008) in Ghana using water to which cadmium and lead had been added to
irrigate cabbage, carrots, and lettuce revealed that cadmium and lead
concentrations increased with irrigation water concentrations significantly with p-
value of cadmium <0.0011 and lead (pb) <0.05.

It has been established that heavy metals in soils will be associated with

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various chemical forms that relate to their solubility which directly bear on
their mobility and biological availability (Xian, 2009).

Heavy metals in soluble form have high relation to their uptake by plants
(Miller and Me free).

Vegetables can absorb metals from soil as well as from deposits on the parts of
vegetables exposed to the air from polluted environments (Haiyan and
Stuanes, 2003). Emission of heavy metals from the industries and vehicles
may be deposited on the vegetable surfaced during their production, transport
and marketing, elevated level of heavy metals in vegetables sold in the market
of Riyadh city in Saudi Arabia due to atmospheric deposition (Al-jassir, 2005).
The extent of absorption of the elements by the plant depends on among other
things, the nature of plant, chemical constituents of the pollutant, concentration
of the element in the soil PH and the interaction with other metals (Zurera 2006).

The uptake and bioaccumulation of heavy metals in vegetables is influenced by

many factors such as climate, atmospheric depositions of heavy metals in soils,
the nature of the soil and the degree of maturity of the plants at the time of the
harvest (Scott and Voutsa, 2017). Air pollution may pose a threat to post .harvest
vegetables during transportation and marketing, causing elevated levels of heavy
metals in the vegetables (Sharma, 2008). Elevated levels of heavy metals in
vegetables will be reported such as long term uses of treated and untreated will
bete water (Sharma, 2006).

2.1 HAZARDOUS EFFECT OF HEAVY METALS ON HUMAN

HEALTH
Heavy metals will be very harmful because of their non-biodegradable nature,
long biological half lives and their potential to accumulate in different body parts

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(Bahamuka and Mubofu, 2007). Most of the heavy metals will be extremely
toxic because of their solubility in water. Nowadays heavy metals will be
ubiqudous because of their excessive use in industries (Jing, 2008).
Factors associated with the possible health effects of exposure to cadmium, lead
and mercury have been investigated over many years in occupational settings,
using experimental animals and human's exposed to environmental pollution.
The type of adverse health effect will be known to a great extent but, because of
the very strong influence of confounding factors, it is very difficult or almost
impossible to find there holds for some out comes such as impairment of
cognitive functions in children exposed to lead or mercury. Chronic low level
intake of heavy metals have adverse effects to human beings and other animals
due to the facts that there is no effective mechanism for their elimination from
the body (Bahamuka and Mubofu, 2018).
Metals such as lead, mercury, cadmium and copper will be cumulative poisons.
These metals cause environmental hazards and will be reported to be
exceptionally toxic (Ellen, 2007). Vegetables take up metals by absorbing them
from contaminated soils, as well as from deposits on parts of the vegetables
exposed to the air from polluted environments (Zurera, 2003).
Prolonged consumption of unsafe concentrations of heavy metals through food
stuffs may lead to the chronic accumulations of heavy metals in the kidney and
liver of humans causing disruption of numerous biochemical processes, leading
to cardiovascular, nervous, kidney and bone diseases (WHO, 2017). Some heavy
metals such as Cu, Zn, Mn, Co and Mo act as micronutrients for the growth of
animals and human beings when present in trace quantities, whereas others such
as Cd, As and Cr acts as carcinogens (Freig, 2014).

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2.2 THE RISK OF CONTAMINANTS ACCUMULATING IN SOIL
ENVIRONMENT AND CROPS
The risk of contaminants accumulating in soil environment and crops due to
sewage water, fertilizer and pollutants is of serious concern. Heavy metals have
been reported to produce mutagenic, teratogenic, neurotoxic and carcinogenic
effect even at very low concentrations (Waalkes, 2009).
Human beings have also been reported to develop several disease like
cardiovascular, tubular dysfunction in kidneys and nervous disorders due to
metal toxicity (WHO, 2017).

2.3 CHILDHOOD EXPOSUER TO HEAVY METALS CAN RESULT IN

LEARNING DIFFICULTIES, MEMORY IMPAIRMENT AND MORE
Memory impairment, and more the young will be more prone to the toxic effects
of heavy metals as the rapidly developing body systems in the fetus, infants and
young children will be far more sensitive, Childhood exposure to some petals
can result in learning difficulties, memory impairment, damage to the nervous
system, and behavioral problems such as aggressiveness and hyperactivity., At
higher doses, heavy metals can cause irreversible brain damage children may
receive higher doses of metals from food than adults, since they consume more
food for their body weight than adults (Jaruf, 2003).
2.4 CURRENT TECHNOLOGIES USED TO DECREASE DIATARY
TOXICITY OF HEAVY METALS
Some micro-organism based remediation techniques, such as bioremediation,
show potential for their ability to degrade and detoxify certain contaminants.
Although these biological systems will be less amenable to environmental
extremes then other traditional methods, they have perceived advantage of being
more cost-effective (Abdullah and Uzairu, 2009, Adam Saleh, 2015).

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Bioremediation can be defined as any process that uses micro-organism, fungi,
and green plants of their enzymes to return the natural environment altered by
contamination ants to its original condition. Bioremediation technologies can be
generally classified as insist or exsit (Abdullah and Uzairu, 2009).

2.5 ACUTE TOXICITY OF COPPER

A cute toxicity of ingested copper is characterized by abnormal pain, diarrhea,
vomiting, tachycardia and a metallic taste in the continued ingestion of copper
compound, can cause cirrhosis and other debilitating liver condition (Mueller,
2021).
Inhaled copper dust or fumes can produce eye and respiratory tract irritation,
headaches, vertigo-drowsiness, chitis, fever, aching muscles and discoloration of
the skin and hair in humans (U.S.A.E, 2008) Vinayard workers exposed to
copper fumes for long period of time developed pulmonary fibrosis and
granulomas of the lungs, liver impairment and liver disease (Cirrhosis, fibrosis
and various morphological changes).
Similar result were obtained in animals chronally exposed to copper containing
dust and fumes (Muellar, 2012). Further animals studies on copper toxicity have
shown varying degrees of liver and kidney damage (necrosis of the kidney;
sclerosis, necrosis and cirrhosis of the liver), decreased total weight, brain
weight and red blood cells count, increased platelet counts and the presence of
gastric ulcers (Rana and Kumar, 2018).

2.6 SYMTOMS OF EXPOSURE TO TOXIC METALS

Exposure toxic metal is associated with many chronic diseases. Recent research
has found that even low levels of lead, mercury, cadmium, aluminum and
arsenic can cause a wide variety of health problems.
I. Decreased intelligence in children

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 II. Nervous system disorders
III. Immune dysfunction
IV. Depression
V. Fatigue
VI. Muscle weakness and aches
VII. Anemia
VIII. Skin rashes
IX. High blood pressure
X. Memory loss
XI. Diarrhea
XII. Nausea
XIII. Metallic taste in mouth
XIV. Irritability
XV. Tremors
XVI. Cancers and
XVII. Hyperactivity

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 CHAPTER THREE

3.0 METHODOLOGY

3.1 MATERIAL
250ml digestion tube

Electronic weighing balance

Timer

What man No: 42 filter paper

25ml volumetric flasks

Test tubes

Spatula

Conical flasks

Pestle and mortar

Polyethylene bag

Oven

Knife/cutters

AAS (atomic absorption spectrophometer) and

Specimen bottles

3.2 REAGENTS
Nitric acid (HNO3)

Perchloric acid (HCIO4)

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Distilled water

3.3 SAMPLE COLLECTION

Samples of vegetables was randomly collected from the farm irritated with
untreated water in Zanna Zakariya housing estate Damaturu. Leafy vegetable was
prepare bed as they accumulate heavy metals in greater capacity than other
vegetables three.

3.4 GLASS WILL BE SAMPLE AND PRE - TREATMENT

All glass and container’s required experimentations were first wash with distilled
water followed by soaking in 10% nitric acid for few hours. Those it was ensured
that no contamination occurred in them. The collected vegetable samples were also
being use with distilled water to remove dust particles. Samples than were cut into
pieces of uniform size and dried. Air dried samples then place in dehydrator for 2-3
days and then oven dried at 1000C.

Now ground into fine powder using pestle and mortar and store in polyethylene
bags until for acid digestion

3.5 NITRIC- PERCHLORIC ACID DIGESTION

Nitric-perchloric acid digestion were performed, by following the procedure
recommended by the A.O.A.C (2019). One gram of sample, was place in a 250ml
digestion tube and 10ml of concentrated HNO3 was added. The mixture were
boiled gently for 30-45 min to oxidize all easy oxidizable matter. After cooling,
5ml of 70% HCIO4 was added and the mixture was boiled gently until dense white
fumes appear was bed. After cooling 20ml of distilled water would be added and
the mixture were boiled further to release any fumes. The solution was cooled,
further filtered through what man NO. 42 filter paper and transferred quantitatively
to 25ml volumetric flask by adding water.

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 CHAPTER FOUR

4.0 HEAVY METAL ANALYSIS

The concentrations of Cd and Cu, in the final solutions was determined by an
atomic absorption spectrophotometer (AAS).

S/ PARAMETER
METHOD #1 #2 #3 MEAN SD
N (Mg/Kg)
1 CADMIUM AAS 0.279 0.28 0.281 0.28 0.001
2 COPPER ASS 0.144 0.144 0.145 0.144 0

4.1 DISCUSSION

The result showed that the levels of copper (cu) ranged from 0.05 to 0.07Ng/g and
0.025 to 0.05 for cadmium (cd) in all the vegetables samples including the soil, the
levels of cadmium and copper varied between the vegetables varieties and from
side to side. The variation in the concentration of the soil, air and irrigation water
of their production sites and also the absorption of heavy metals from areal
deposition during transportation and marketing. There was a significant different
(p<0.05) in the levels of cadmium between the vegetable varieties. The levels of
both cadmium and copper in the vegetables are found to be below the maximum
permissible levels recommended by FAO (WHO) for the two metals in the
vegetables.

Vegetable can absorb metals from soil as well as from deposited on the part of the
vegetables exposed in to the air from polluted environment (Haiyand stuance,
2003) the Zanna Zakariya farm soil in the table (4.0) has the highest concentration
of copper (cu).

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 CHAPTER FIVE
5.1 CONCLUSION

From this study, it may be conclude that monitoring and assessment of heavy
metals concentrations in different vegetables from the production as well as market
sites required more extensive studies for assessing the risk of health hazards to the
human beings. The present study further suggested that to reduce the consumption
as washing can removed a significant amount of areal contamination from the
vegetable surface.

5.3 RECOMMENDATION

I. To reduce the health risks vegetable should be washed properly before

consumption as a washing can removed a significant amount of aerial
contamination from the vegetables surface.
II. Appropriate percussions should be taked at the time of planting.
Transportation and marketing.
III. Since the intake of heavy metals can lead to altering of human and animals
healthiness state. It is therefore suggested that regular monitoring of heavy
metals in the plant tissue is essential in other to prevent excessive build-up
of these metals in the human foods chain.
IV. Munch study is needed in this respect such as metal update studies at
cellular afflux of organelles membranes.

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DETRA and environment agency (2002) assessment of risk to human health from
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Y.Z and Zhu Y.G (2008) health isk of heavy metal in contaminat soil and food
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Khang,S., Cao, Q.,Zheng, Y.M., Hueng,Y.Z. and Zhu,Y.G.(2008) Health

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Marshal, (2004) Enhance Food chain intergrity: Quatity Assurance Machanism for
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Zhuang, P. Yang, Q.W.H.B Wang and Shu, W.S. (2007) Phyto Extraction of
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Freig, D.I. Reid, T.M. and Loeb, L.A.(2000) Reactive Oxygen Species in
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 APPENDIX
Conversion of A.A.S result to Ng/g
Dilution Factor=volume of soluent used / number of gram of sample
Zanna Zakariya soil analysis for copper (cu) Dilution Factor x Absorbance
25x0.003=0.075 ug/g
Cadmium (cd)
25x0.01=0.025 ug/g.

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