CHAPTER ONE

1.0 Introduction

1.1 Background of the study

In recent years, significant attention has been paid to the problem of environmental

contamination by a wide variety of chemical pollutants including heavy metals (Isola et al.,

2015). These chemicals arise from increasing levels of anthropogenic activities such as

industrialization and urbanization, coal and metals ore mining, chemical manufacturing,

petroleum mining and refining, electric power generation, melting and metal refining, metal

plating and to some extent domestic sewage. Little attention has been paid to the tree as one of

the major sources of heavy metals pollution in the environment (Adewole & Uchegbu 2010;

Abidemi 2011; Okoro et al. 2013). Odiwe et al. (2014) has also worked on heavy metals

pollution.

Over the years, man has relied so much on medicinal plants for health and food needs

(Nwachukwu et al, 2010). The traditional uses of medicinal plants for curing and preventing

illnesses, including the promotion of both physical and spiritual well-being of humans are well

documented (Idu & Onyibe, 2007). The use of medicinal herbs to relieve and treat many human

diseases is increasing around the world due to their mild features and low side effects (Yap et al,

2010).

1
World Health Organization (WHO, 2007) survey indicated that about 70–80% of the world

population relies on non-conventional medicine, mainly of herbal sources, in their primary health

care. This report revealed that medicinal plants and their trace elements play an important role in

the treatment of diseases (Chan, 2003). Heavy metals are known to bio accumulate and thus

disrupt functions of vital organs and glands in the human body, affecting brain, kidney and liver

(Suranjana & Manas, 2009). Medicinal plants can be contaminated by these heavy metals via

root uptake or by direct deposition of contaminants from the atmosphere onto plant surfaces

(Dzomba et al., 2012; Olowoyo et al, 2011). It is recommended that herbal drugs should not be

used without qualitative and quantitative analysis of their heavy metal contents. In an attempt to

control pollution of medical plants by heavy metal World Health Organisation (WHO, 2007),

regulated maximum permissible limits of heavy metals like arsenic, cadmium and lead to 0.1,

0.3, 5.0 mg/kg, respectively (WHO, 1998; 2007). Ingestion of high concentration of heavy

metals such as chromium, cadmium and lead can affect growth rate, cause hepatic and renal

impairment (Blowes, 2002).

Moringa oleifera (drumstick tree), popularly known in many countries as a “miracle plant”

because of its nutritional and medical properties has a great bio-energy potential and is an

exceptionally nutritious vegetable tree with many uses (Ozumba 2008; Onyekwelu &

Olabiwonnu, 2010). Moringa is rich in many vitamins, including vitamin A, several forms of

vitamins B, C, D and E. In fact, it has more of these vitamins than many varieties of foods (such

2
as carrots, oranges and milk) and is known to be excellent source of these vitamins (Zarkada et

al., 1997). Moringa leaves are edible and they form part of traditional diets in many countries

(Odee, 1998). The leaves are rich in protein, vitamins and minerals (Fuglie, 2006). Despite the

aforementioned benefit of Moringa oleifera, little is known about its bioaccumulation potentials

in the face of ever increasing environmental contamination.

1.2 Aims and Objectives of the study

1.2.1 Aim

The aim of the study is to determine the bioaccumulation of heavy metal in Moringa oleifera

trees.

1.2.2 Objectives

Objectives of the study includes:

i. Collection of Moringa oleifera leaves, root and stem from different locations.

ii. Collection of the soil where he tree is located.

iii. Evaluate the heavy metals present in the soil, he leaves, the root and stem

1.3 Justification

Moringa oleifera has been used for centuries due to its medicinal properties and health benefits.

However, the tree is found in various places including dumpsite because it is domesticated.

Therefore, evaluating bioaccumulation of heavy metals is important to assess the safety of use of

the different part of the tree Moringa oleifera

3
 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Composition of Moringa oleifera

The composition of Moringa oleifera varies depending on climatic variations, crop management,

whether it is cultivated or wild, the state of maturity of the plant at the time of harvesting, the

type of post-harvest processing and depends on the growing area, i.e., the land where it is grown

2.1.1 Primary Metabolites in Moringa oleifera

Moringa oleifera leaf is a rich source of minerals, such as calcium, potassium, zinc, magnesium,

iron, phosphorus and copper. One of the characteristics of the leaf is its high protein content, due

to the essential amino acids, which constitute about 30% of its weight, being comparable to milk

powder which contains 35%, and is available all year round, as the protein and essential amino

acid content is present in the leaves, unlike other plants which contain them in the seeds,

reporting a protein content of 29.4 g protein/100 g dry weight in the leaves. Thus, Moringa

oleifera can be considered a new source of protein to be included in food, like chia seed, with a

protein content of 24 g protein/100 g dry weight (Amaglo and Bennett, 2011). Regarding

carbohydrates, its level is lower (8.1%) compared to the other parts of the plant. In addition,

fibres were also found, with a value ranging between 18.1 and 21.1 g/100 g dry weight of the

leaves (Amaglo and Bennett, 2011).

4
The leaves are noted for high levels of β-carotene and provide more vitamin A than carrots and

pumpkin, however it is not clear whether this vitamin is retained even after drying and grinding

the plant. Even so, studies have shown that their consumption is sufficient to counteract the

effects of this vitamin deficiency. They are also a good source of B vitamins (Leone and Spada,

2015), among which thiamine, riboflavin and niacin have been found, with a concentration

between 0.06 and 0.6 mg/100 g, 0.05 and 0.17 mg/100 g and 0.8 and 0.82 mg/100 g for thiamine,

riboflavin and niacin, respectively. In the dried leaf, their concentrations were 2.85, 22.16 and

8.86 mg/100 g DW, respectively (Leone and Spada, 2015).

On the other hand, supplementation with 100 mg/dL of Moringa oleifera leaf per day has a

similar effect to treatment with vitamin E at 50 mg/dL per day(Leone and Spada, 2015).

Contains more vitamin C than an orange and more calcium than dairy products, however a

significant part of this calcium is present in the form of calcium oxalate crystals, which cannot be

used by the body and is eliminated directly without being absorbed. In addition, Moringa

oleifera is high in potassium and iron; even more than bananas and spinach respectively.

Moringa oleifera seed has a high proportion of monounsaturated/saturated fatty acids

(MUFA/SFA), sterols and tocopherols, as well as proteins rich in sulphur amino acids (Leone

and Spada, 2015).

5
2.1.2 Secondary Metabolites of Moringa oleifera

The different parts of Moringa oleifera are good sources of glucosinolates, flavonoids and

phenolic acids, carotenoids, tocopherols. Alkaloids, saponins, tannins, steroids, phenolic acids,

alkaloids, carotenoids, polyphenols, isothiocyanates, phytates, glucosinolates, flavonoids and

terpenes can be found in the Moringa oleifera leaf. Among the glucosinolates, benzyl 4-O-(α-L-

rhamnopyranosyloxy)-glucosinolate is the most predominant (glucomoringin). Its leaves include

11 phenolic acids (gallic acid, caffeic acid, chlorogenic acid, ocoumaric acid, p-coumaric acid,

ellagic acid, gentisic acid, sinapic acid, syringic acid) and their derivatives (coumaroylquinic

acids and their isomers, feruloylquinic and caffeoylquinic), 26 flavonoids (present mainly as

flavonol and glycoside: quercetin, rhamnetin, campferol, apigenin and myricetin. Flavonoids

include flavonol glycosides (glycosides, rutinosides and malonylglycosides) of quercetin

“kaempferol” 0.05–0.67%) isorhamnetin and lignans (isolariciresinol, medioresinol,

epipinoresinol glycosides and secoisolariciresinol) (Kumar and Singh, 2012).

The flavonoid composition is higher in the leaves than in the seeds, ranging from 2000 to 12,200

mg per dl of Moringa oleifera leaf. Moringa oleifera seed contains phytosterols, the most

abundant of which are β-sitosterol, stigmasterol and campesterol. Alkaloids, saponins, phytates,

tannin and phenolic compounds (quercetin and p-hydroxybenzoic acid) can also be found. The

seed is oleaginous and has aleurone sources with a lectin fraction, is an oil that must be refined to

be consumed and contains a similar composition to oleic acid, however, the composition may

6
vary according to the geographical location of the plant. Seeds are a good source of

glucosinalates (8–10%), glycosylate isothiocyanate, 4-(α-Lrhamnosyloxy) benzyl ITC and 4-O-

(α-L-rhamnopyranosyloxy)-benzylglucosinolate (glucomoringin). In other edible parts of

Moringa oleifera, more than 102 bioactive compounds have been identified in the root, while 74

essential oils have been identified in the flowers. In addition, both the peel and the dried pod of

Moringa oleifera fruit contain high levels of polysaccharides and glucans; 28% in the peel and

32% in the pod (Amaglo and Bennett 2014). Glucosinolates have also been found, of which 4-O-

(α-L-rhamnopyranosyloxy)-benzylglucosinolate (glucomoringin) is the most predominant in the

stem, flowers and pods of Moringa Oleifera (Amaglo and Bennett 2014).

Although in the roots, benzyl glucosinolate (glucotropaeolin) is the most prominent, flavonoids,

notably flavonol glycosides (glycosides, rutinosides, and malonylglycosides) of quercetin

[kaempferol isorhamnetin] are also present in various parts of the plant, except in the roots and

seeds, since this plant organ is the most studied and most consumed, which concentrates most of

the nutrients (Ma et al, 2011).

2.2 Biological Effects of Moringa oleifera

The bioactive compounds (Figure 1) present in Moringa oleifera confer properties associated

with disease prevention and treatment, such as antimicrobial, anti-inflammatory, anticancer,

antidiabetic, antioxidant, hepatoprotective and cardioprotective. Primary and secondary

metabolites may also be involved in these applications. Primary metabolites are proteins,

7
polysaccharides and lipids involved in physiological functions. Among them, polysaccharides

and fibres are the main compounds showing positive effects on chronic diseases such as cancer,

cardiovascular diseases, diabetes and obesity. On the other hand, secondary metabolites are

minor molecules, such as phenolic compounds, halogenated compounds, sterols, terpenes and

small peptides. Most of the phytochemicals reported in Moringa oleifera offer potential in the

prevention and treatment of diseases. The anti-inflammatory effect is due to the content of

flavonoids, alkaloids, tannins and glycosides, among which quercetin appears to inhibit NF-KB

activation, producing an antiinflammatory effect (Das and Sikder, 2011).

Other compounds with an anti-inflammatory effect include kaempferol derivatives, flavonol

glycosides, aurantiamide acetate, 1,3-dibenzylurea, diterpenes, α- and β-amyrin, benzaldehyde 4-

0-β-glucoside, β-sitosterol, rutin, and glucosinolate, mainly attributed to the glycosylate

isothiocyanate, 4-(α-L-rhamnosyloxy) benzyl ITC, resulting from myrosinase. Moringa oleifera

reduces inflammation by suppressing inflammatory enzymes and proteins in the body, and leaf

concentrate can significantly reduce inflammation in cells (Chaudhary, 2013).

The antimicrobial effect provided essential oils from the leaves and alcoholic extracts of the

seeds. In fact, Chuang et al, (2012) demonstrated this activity of the leaf and leaves against

dermatophytes such as Trichophyton rubrum and Trichophyton mentagrophytes. In addition to

these compounds, other compounds have been found that also produce this effect, 4(βL-

rhamnosyloxy) benzyl isocyanate or pterigospermine,4- (β-D-glucopyranosyl-1→4-β-l-

8
ramnopyranosyloxy),benzyl thiocarboxamide,(-)-Catechin, phenylmethanamine, 4β-D-

glucopyranosyl-1–>4 β-L-rhamnopyranosyloxy)-benzyl iso- Plants cyanate, niazirine and

glucosinolate mainly attributed to the glycosylate isothiocyanate, 4-(α-L-rhamnosyloxy) benzyl

ITC, resulting from myrosinase. Phenolic compounds have been associated with the

antimicrobial and antifungal activities of Moringa oleifera extracts, the leaves being the organs

with the highest amount of these compounds. Regarding the antimicrobial effect of Moringa

oleifera plants when included in food, Moringa Oleifera contributes to control the growth of

undesirable microorganisms, due to low pH values and the presence of pterigospermin. The roots

of Moringa oleifera have antibacterial properties and are described to be rich in antimicrobial

agents. The bark extract has been found to have antifungal activities, while the juice of the bark

and stem show an antibacterial effect against Staphylococcus aureus (Khor and Lim, 2018).

Studies have shown the anticarcinogenic effect of several compounds, namely glycosylated

isothiocyanate, benzyl carbamate niazimycin and β-sitosterol, which have anti tumour properties

against lung, breast, skin, oesophageal and pancreatic cancer. These compounds are found in

high concentrations in the leaves and seeds of the plant. Moringa oleifera is rich in ascorbic acid,

which provides an anti-diabetic effect by aiding insulin secretion, and another compound found

in Moringa Oleifera that produces this effect is myricetin. Antioxidants are popular because they

scavenge free radicals that cause oxidative stress, cell damage and inflammation. Moringa

9
oleifera contains antioxidants called flavonoids, polyphenols and ascorbic acid in the leaves,

flowers and seeds (Farooq, 2011).

Furthermore Studies have shown that the plant is rich in polyphenols, which gives it a high

antioxidant capacity. The compounds in Moringa oleifera that provide this activity are feluric,

gallic and ellagic acids, β-sitosterol, myricetin, niazimycin, niacimicin A and B, tocopherols: α-

tocopherol, δtocopherol, γ-tocopherol, vanillin, kaempferol, quercetin, β-carotene (-)-catechin,

astragalin and isoquercetin. Moringa oleifera plays an important role in protecting the liver from

damage, oxidation and toxicity due to the high concentrations of polyphenols in its leaves and

flowers. Moringa oleifera oil can also restore liver enzymes to normal levels, reducing oxidative

stress and increasing protein content in the liver.

The flowers and roots of the Moringa oleifera plant contain a compound called quercetin, which

is known to protect the liver. Other compounds contained in the plant with this activity are β-

sitosterol, quercetin and some of its glycosides, rutin and flavonoids, which also prevent lipid

oxidation. Moringa oleifera leaves and seeds have been found to help lower blood pressure; this

is due to compounds called glycosides, and in the leaves it is also due to N-α-

Lrhamnophyranosyl vincosamide. Moringa oleifera leaf extract has also been found to

significantly reduce cholesterol levels due to the action of β-sitosterol (Farooq, 2011).

10
2. 3 Nutrient contents of Moringa oleifera

The most amazing fact about Moringais that it is a storehouse of nutrients and medicinal

chemicals.As it was reported by different scholars (Jongrungruangchok, 2010), Moringa tree is

rich in nutrients such as minerals, fiber and proteins that can play essential role in human

nutritional consumption. Numerous of the research reports have shown that Moringa

oleifera leaves has high protein compared to with other leaves eaten as food.

A recent research on dietary iron supplements and Moringa oleifera leaves influence the liver

revealed that iron from Moringa oleifera can overcome iron deficiency (Saini, et al, 2014) and

Similarly, research done on the relative bioavailability of folate from the traditional food

plant Moringa oleiferaindicate that the relative bioavailability of folate from Moringa

oleifera leaves using rat model was very high and thereforeMoringa oleifera leaves can be a

potential source of dietary folate (Saini, et al, 2014).

Numerous research reports on the medicinal and nutritional of Moringa oleifera now are existent

in scientific journals and the widespread literature. Moringa use as medicinal and nutritional

purposes was started since centuries. (Mahmood et al, 2010). Moringa oleifera contains all the

essential medicinal and nutritional properties and anextremely valuable food source that are vital

for human and livestock consumptions (Mahmood et al, 2010). Study on the potential uses

of Moringa oleifera by (Rockwood et al, 2013)

11
2.4 Benefits/uses of Moringa

There are many uses of Moringa tree and these will all be: medicines, Human food, Water

purification, Animal fodder, Alley cropping, Fertilizer, Living fence, Living fence, Domestic

cleaning agent, Fuel wood and other uses. Moringa increased physical energy - Tune the body up

with naturally occurring nutrients to make the energy last longer. Numerous research reports

reveal that, parts of Moringa plant can be used in different techniques. Moringa oleifera seed and

leaves is advantageous source of nutrients, medicines, clean dirty water and it can be used for

alley cropping; because, it has lots of leafy material. The uses Moringa oleifera are well

documented by Fahey, as nutritional, industrial, medicinal, and agricultural advantage

Moringa oleifera has great potential for prevention of different diseases like nutrient deficiency,

cancer, anemia swell as for dirty water purification. Moringa powder contains sufficient amount

of vitamins, nutrients and chemicals in it. This makes the tree a medicine for many different

diseases. Moringa oleifera has also promoted by World Health Organization (WHO) as an

alternative to imported food source to treat malnutrition (Utalangka et al, 2013).

2.4.1 Medicinal benefits/uses of Moringa

Moringa has long been used in herbal medicine by Africans and Indians. Moringa oleifera is

often referred as a panacea and can be used tocuremorethan300diseases. In this section,

medicinal uses of Moringa leaves are reviewed. Different studies shown that, all parts

12
of Moringa oleifera (leaves, fruits, immature pods, and flowers) are incorporated into the

traditional food of humans in many tropical and subtropical countries.

2.4.2 Medicinal uses of all parts of Moringa

Various parts of Moringa tree act as cardiac and possess antitumor, circulatory stimulants,

antiepileptic, antipyretic, antiulcer, anti-inflammatory, antispasmodic, anti-hypertensive,

diuretic, antioxidant, anti-diabetic, cholesterol lowering, antibacterial, hepatoprotective and

antifungal activities.

Leaves: Moringa leaves are the most commonly used part of the plant. Moringa leaves treathy

perglycemia, asthma, flu, heart burn, Dyslipidemia, malaria, syphilis, diarrhea, pneumonia,

scurvy, headaches, bronchitis, skin diseases, eye and ear infections. Also reduces, blood

pressure and cholesterol and acts as an anticancer, Antioxidant, anti microbial, anti-

atherosclerotic and ant diabetic agents, neuro protectant (Utalangka et al, 2013).

Seeds: Numerous researches reported that, Moringa seed powder can be used as for cleaning

dirty water in a simple and quick method. Seeds of Moringa help in treating Chrohn’s disease,

hyperthyroidism, rheumatism, antiherpes-simplex virus arthritis, cramp, epilepsy gout, and

sexually transmitted diseases, can act asantimicrobial and anti-inflammatory agents (Utalangka

et al, 2013).

13
2.5 Other Sources of heavy metals

In Nigeria, numbers of Moringa oleifera have increased mainly due to large inflow of used

Moringa oleifera into the country. These Moringa oleifera are usually not registered but assist

low income earners patronizing and as such contributed remarkably to the problem of soil

contamination in most cities in the country (EEA, 2007). Most of these dispose their waste oil

and fossil fuel among others into their nearby soil, stream or river thereby polluting the

environment. The indiscriminate disposal of these waste engine oil into gutters, water drains,

open vacant plots, and farm land is a common practice in Nigeria, and this has led to a significant

environmental pollution (Akinpelumi & Olatunji, 2015).

These wastes engine oil contain heavy metals such as Cd, Cr, Pb, and Ni, Zn, Tl, Cd, Cu, (Elena

et al., 2004, Ilemobayo & Kolade 2008). When the environment is contaminated with these

heavy metals, organisms tend to absorb these toxic substances (heavy metals or chemicals) at a

rate faster than that at which the substance is lost. (Bryan & Waldichuk, 2012). Plants

accumulate minerals essential for their growth from the environment and may accumulate some

metals which have no known direct benefit to the plants (Ademoroti, 1986). However, the rate at

which various species of wild plants growing in the same habitats accumulate heavy metals may

vary considerably (Byliñska, 1995). Detailed ecological studies enabled the distinction of species

with particular accumulative capacities with respect to one or several heavy metals, thereby

serving as bioindicators of contaminated areas (Holoubek et al, 2000). Research findings by

(Divrikli et al. 2006) have shown that the concentration of essential elements in plants is

conditional; as it is affected by the characteristics of the soil and the ability of plants to

selectively accumulate some of these metals.

14
2.6 Heavy metals of Moringa oleifera

Heavy metals refer to as any metallic chemical element that has a relatively high density and is

toxic or poisoinous at low concentration example of heavy metals include Aluminum (Al),

Arsenic (As), Thallium (Tl) and Lead (Pb).

2.6.1 Aluminium

Aluminium (aluminum in American and Canadian English) is a chemical element with the
symbol Al and atomic number 13. Aluminium has a density lower than those of other common
metals, at approximately one third that of steel. It has a great affinity towards oxygen, and forms
a protective layer of oxide on the surface when exposed to air. Aluminium visually resembles
silver, both in its color and in its great ability to reflect light. It is soft, non-magnetic and ductile.
It has one stable isotope, Al; this isotope is very common, making aluminium the twelfth most
common element in the Universe. The radioactivity of Al is used in radiodating (Divrikli et al.
2006).

2.6.2 Titanium

Titanium is a chemical element with the symbol Ti and atomic number 22. Its atomic weight is
47.867 measured in daltons. It is a lustrous transition metal with a silver color, low density, and
high strength. Titanium is resistant to corrosion in sea water, aqua regia, and chlorine. Titanium
was discovered in Cornwall, Great Britain, by William Gregor in 1791 and was named by Martin
Heinrich Klaproth after the Titans of Greek mythology. The element occurs within a number of
mineral deposits, principally rutile and ilmenite, which are widely distributed in the Earth's crust
and lithosphere; it is found in almost all living things, as well as bodies of water, rocks, and soils.
The metal is extracted from its principal mineral ores by the Kroll and Hunter processes. The
most common compound, titanium dioxide, is a popular photocatalyst and is used in the
manufacture of white pigments.Other compounds include titanium tetrachloride (TiCl4), a
component of smoke screens and catalysts; and titanium trichloride (TiCl3), which is used as a
catalyst in the production of polypropylene (Divrikli et al. 2006).

15
2.6.3 Zinc

Zinc is a chemical element with the symbol Zn and atomic number 30. Zinc is a slightly brittle
metal at room temperature and has a silvery-greyish appearance when oxidation is removed. It is
the first element in group 12 (IIB) of the periodic table. In some respects, zinc is chemically
similar to magnesium: both elements exhibit only one normal oxidation state (+2), and the
Zn2+ and Mg2+ ions are of similar size Zinc is the 24th most abundant element in Earth's crust and
has five stable isotopes. The most common zinc ore is sphalerite (zinc blende), a zinc
sulfide mineral. The largest workable lodes are in Australia, Asia, and the United States. Zinc is
refined by froth flotation of the ore, roasting, and
final extraction using electricity (electrowinning). Brass, an alloy of copper and zinc in various
proportions, was used as early as the third millennium BC in the Aegean area and the region
which currently includes Iraq, the United Arab Emirates, Kalmykia, Turkmenistan and Georgia.
In the second millennium BC it was used in the regions currently including West
India, Uzbekistan, Iran, Syria, Iraq, and Israel. Zinc metal was not produced on a large scale until
the 12th century in India, though it was known to the ancient Romans and Greeks. The mines
of Rajasthan have given definite evidence of zinc production going back to the 6th century
BC. To date, the oldest evidence of pure zinc comes from Zawar, in Rajasthan, as early as the 9th
century AD when a distillation process was employed to make pure zinc. Alchemists burned zinc
in air to form what they called "philosopher's wool" or "white snow" (Prasad et al, 2014).

2.6.4 Copper

Copper is a chemical element with the symbol Cu (from Latin: cuprum) and atomic number 29.
It is a soft, malleable, and ductile metal with very high thermal and electrical conductivity. A
freshly exposed surface of pure copper has a pinkish-orange color. Copper is used as a conductor
of heat and electricity, as a building material, and as a constituent of various metal alloys, such
as sterling silver used in jewelry, cupronickel used to make marine hardware and coins,
and constantan used in strain gauges and thermocouples for temperature measurement.

Copper is one of the few metals that can occur in nature in a directly usable metallic form ( native
metals). This led to very early human use in several regions, from c. 8000 BC. Thousands of
years later, it was the first metal to be smelted from sulfide ores, c. 5000 BC; the first metal to be

16
cast into a shape in a mold, c. 4000 BC; and the first metal to be purposely alloyed with another
metal, tin, to create bronze, c. 3500 BC.

In the Roman era, copper was mined principally on Cyprus, the origin of the name of the metal,
from aes сyprium (metal of Cyprus), later corrupted to сuprum (Latin). Coper (Old English)
and copper were derived from this, the later spelling first used around 1530 (Holoubek et al,
2000).

17
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Materials

3.1.1 Sample Collection

The sample used for the analysis was collected from Ado-Ekiti and Ido-Ekiti. The coordinate of

the spot where the sample was taken was obtained having the coordinate of (7.620326 0 N,

5.250450 E) and (7.5077°N, 3.7194°E). The leaves of the tree was collected and the bark of the

stem was peeled out. To collect the root sample of the tree, the root was dug down for about 2ft

deep in the soil before cutting. The soil sample collection was from 5 different points around the

tree. The soil was dug and the soil deep down the hole was obtained and then mixed together

before the sample for analysis was taken. The sample used for the analysis was authenticated by

a botanist at the Ekiti State University Ado-Ekiti (EKSU) with the herbarium number UHAE

2022004.

3.1.2 Sample Pretreatment

After collection of each samples needed, they were dried in the oven. After drying of the sample,

it was then crushed and grinded into finer particles for easy digestion.

3.1.3 Chemicals and Equipment

All reagent used were of analytical grade. Other equipment is ICP-OES 720, fume cupboard, hot

plate/heater, weigh balance and computer.

3.2 Digestion Method

2.5 gram of the sample each was measured out using the weighing balance into a conical flask

and 10 ml of Aqua regia i.e. (3:1 Conc. HNO3:HCL) was added to the sample inside the conical

flask each, the mixture inside the conical flask was placed on the hot plate inside the fume

cupboard for the extraction of the gas coming out of the sample while boiling because it is a

18
poisonous gas that shouldn’t be inhaled. The mixture was digested for about 1 hour at 200 0C

using a heater, during heating of the sample, a clear fume and a clear solution was observed and

the sample was removed from the hot plate in the fume cupboard and was allowed to cool

naturally. After cooling process was completed, digestate was approximated 5ml for each sample

was then filtered into beaker using the whatman filter paper and the funnel and the filtrate was

diluted to 20ml using ultra-pure de-ionized water for ICP-OES analysis.

3.3 Analytical Calibration Procedure For ICP-OES

An appropriate concentration range of the working standards from the multi-element stock

standard was prepared through serial dilution method. A method for creating a new worksheet or

an opening template from the ICP-OES was developed by an expert software using an

appropriate conditions and parameters for fitting the calibration curve on the worksheet a set was

created. The element of interest alongside with their suitable wavelength for their measurement

was selected and the method to be used, input sample matrix, blank, standards, and sample codes

was created and the method created was then saved. The blank and the standards was inputted in

the appropriate rack position on the auto-sample with the blank read first, then the working

standards. The calibration coefficient was checked and the regression equation of R 2 ≥ 0.995 was

ensured to be followed. (If standard lower than this value, recalibrate the instrument). The

sample codes for the samples to be analyzed was entered on the Sequence page and the analysis

can began, after analyzing the end of the sample run, the blank and the rinse-solution was ran for

few minutes with the result obtained saved. After 5 minutes rinse of the instrument, it was then

powered off.

19
 CHAPTER FOUR

4.0 RESULT AND DISCUSSIONS

4.1 RESULT

Table 4.1: Mineral concentrations of the different parts of Moringa oleifera samples

Elements Samples

Soil Root Stem Leaves

A B A B A B A B

Al 0.1031±0.10 0.3416±0.02 1.3844±0.12 1.3931±0.18 1.3618±0.15 2.6558±0.34 2.6558±0.34 2.0037±0.55

As 0.0448±0.01 0.1987±0.02 0.0650±0.00 0.0944±0.02 0.1789±0.01 0.1000±0.02 0.1152±0.03 0.1725±0.01

Cd -0.0543±0.00 -0.2209±0.00 -0.2260±0.00 -0.2227±0.00 -0.2112±0.00 -0.2209±0.00 -0.2250±0.00 -0.2242±0.00

Cr 0.2834±0.00 1.0629±0.00 -0.2344±0.00 -0.2014±0.00 -0.1705±0.00 -0.2112±0.00 -0.1945±0.00 -0.2046±0.00

Cu 0.1708±0.00 0.7399±0.00 -0.2225±0.00 -0.1849±0.00 -01829±0.00 -0.2063±0.00 0.1692±0.00 0.1778±0.00

Ni 0.0921±0.00 0.3023±0.01 -0.1060±0.00 -0.1050±0.00 -0.0638±0.00 -0.0978±0.01 -0.1132±0.01 -0.1069±0.00

Pb 0.0291±0.01 0.8636±0.01 -0.1136±0.00 -0.0747±0.01 -0.0255±0.01 -0.0711±0.00 -0.0465±0.02 -0.0974±0.01

Se 0.0121±0.03 0.1957±0.01 -0.1222±0.00 -0.3000±0.00 -0.0952±0.03 -0.0357±0.02 -0.0±0.02 -0.0085±0.02

Tl 0.0460±0.01 0.2306±0.01 0.1912±0.01 0.0476±0.04 0.1008±0.04 0.2131±0.01 0.2104±0.02 0.1988±0.02

Zn 0.1935±0.00 0.6943±0.00 0.1048±0.00 0.1224±0.00 0.0144±0.01 0.0524±0.00 0.1792±0.00 0.1448±0.00

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Table 4.2: Bioaccumulation factors of the different minerals

Element/Label Bioaccumlation factor for sample A Bioaccumulation factor for Sample B

Al 13.2085 7.7746

As 3.9933 0.5033

Cd - -

Cr - -

Cu - -

Ni - -

Pb - -

Se 7.8867 2.3959

Ti - -

Zn - 0.7034

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Concentration of heavy metal in Moinga oleifera samples are shown on Table 4.1. The result
showed that Pb was not present in any of the sample from both locations, while Se was present in
all the locations from both locations. The root sample from both location did not contain Cd, Cr,
Cu, Ni, Pb and Tl while sample A had Al and sample B had Zn. The stem samples from both
locations did not contain Cd, Cr, Cu, Ni, Pb and Tl while sample A and sample B contain Al, and
sample B contain Zn. The soil sample from both locations contain Al, Cd, Cr and Zn while
sample B had Cr and Cu the leaf sample from both locations did not contain Cd, Cr, Cu, Ni and
Co, while sample from both location had Zn and sample A had Al. Comparing the
Bioaccumulation factors (Table 4.2) the result showed significant factors for As (0.5033) and Zn
(0.7034) from sample B. Others calculated were not significant while many others could not be
calculated because they were either absent in one or both of soil and stem samples.
4.2 DISCUSSIONS

The concentration of heavy metal in Moringa oleifera plant, leaves and soil sample are shown in
Table 4.1 and they include Al, As, Cd, Cr, Cu, Ni, Pb, Se, Tl, Zn. These result revealed that the
Moringa leaves collected from the study area contain appreciable amount of heavy metals.
Heavy metal in different part of the Moringa plant exhibited variation.

Aluminium content in the stem sample (Table 4.1) is generally high and in the range of 2.6558 +
0.434 and 1.3618 + 0.15 (Aluminium sample A and sample B respectively). These were lower
than aluminium reported by (Alfzal et al, 2011). The concentration of other heavy metals are in
moderate range as compared with the limit set by World health organization (WHO, 2007).

The result of the Bioaccumulation factor of the Moringa plant and soil samples is shown in Table
4.2 shown significant bioaccumulation factors lower than 1 signifies that such metal is
bioaccumulated in the plant (Alfzal et al, 2011). These means that As and Zn are bioaccumulated
in the tree from where sample B was collected

Zinc an essential component of various enzymes that are responsible for driving many metabolic
reactions in all crops. Zinc is least toxic and an essential element in human diet as it is required
to maintain the function of the immune system, Zn deficiency in the diet may be highly
detrimental to human health than too much zinc in the diet (ATSDR, 2007).

22
Arsenic mineral has the most abundant translocation factor in the soil and has one of the most
available bioaccumulation values in the sample plant. Research have shown that much level of
arsenic in the soil will absorb to particulate matter and sediment and generally induces he
production of reactive oxygen species that can lead to the production of antioxidant metabolites
and numerous enzymes involved in antioxidant defense (Cao et al, 2014)

All concentration of heavy metal in Moringa plant sample were within. The recommended daily
allowance (RDA) limits (ATSDR, 2007). RDA, are average daily dietary intake level is expected
to be sufficient to meet the nutrient requirement for all heavy individual. The result, therefore,
would suggest that consumption of Moringa plant provide users when taken as it can also be use
as herb supplement base on the knowledge acquired through research work.

23
 CHAPTER FIVE

5.0 CONCLUSION AND RECONMENDATION

5.1 Conclusion
Moringa oleifera sample B contain more quantity of the heavy metals analyzed than sample A.

Also, tree from where sample B is collected bioaccumulates Zn and As.

5.2 Recommendation
Moringa oleifera plant has been known to have many nutritional and medicinal values when

taken. It is also being use as an herb supplement. Therefore consumption of Moringa oleifera

collected from the tree in a dump site is discouraged.

24
 References

Abidemi O. O. (2011). Levels of Pb, Fe, Cd and Co in Soils of Automobile Workshop in Osun

State, Nigeria. Journal of Applied Sciences and Environmental Management 15(2): 279–

282.

Akinpelumi B. E. and Olatunji O. A. (2015). Effects of sawdust soil amendment on the soil,

growth and yield of Solanum esculentum Linn. in waste engine oil-polluted soil.

Sciences in Cold and Arid Regions 7(2): 128–136.

Alfzal. and Abegunde K. D. (2011). Heavy metals contamination of soil and groundwater at

automobile mechanic villages in Ibadan, Nigeria. International Journal of the Physical

Sciences 6(5): 1045–1058.

Amaglo, H. M., Benneth. and Quarmby C. (2011). Chemical analysis o ecological materials.

ATSDR (2007) All concentration of heavy metals in Moringa plant sample were within.

Blowes D. (2002). Tracking hexavalent chromium in groundwater. Science 295: 2024–2025.

Cao (2014) Production of reactive oxygen species that can lead to the production of antioxidant

metabolites.

Chan (2003) Heavy metals are known to bio accumulate and thus disrupt functions of vital

organs and glands in the human body.

Chuang (2012) The antimicrobial effect provided essential oils from the leaves and alcoholic

extracts of the seeds.

25
Divirikli (2006) The radioactivity of aluninium is used in radiating.

Dzomba (2012), Olowoyo (2011) Herbal drugs should not be usede without qualitative and

quantitative analysis of their heavy metals.

Farooq (2011). Properties of Soils and plants uptake within the vicinity of selected Automobile

workshops in Ile-Ife Southwestern, Nigeria. Ethiopian Journal of Environmental Studies

and Management 3(3) 31– 35.

Fuglie (2006) the leaves are rich in protein, vitamins and minerals

Isola (2015) environmental contamination by a wide variety of chemical pollutants including

heavy metals.

Kurma & Singh (2012) flavonol glycosides (glycosides, rutinosides and malon yllcosides), of

quercetin epipinoresinol glycosides and secoisolariesinol.

Ma (2015) flavonol glycosides (glycosides, rutinosides and malon yllcosides), of quercetin

epipinoresinol glycosides and secoisolariesinol.since this plan oan is the most studied

and most consumed.

Nwachukwu (2010) The tradition uses medicinal plant for curing and preventing illness

Olabiwonnu (2010). Environmental Chemistry and Toxi Ecology, Folulex Press Ltd. Ibadan.

Onyinbe (2007) the use of medicinal herbs to relieve and treat many human diseases is

increasing around the world.

26
Prasad (2014) Alchemiss uned zinc in air o om what they called philosophe’s wool or white

snow.

Rockwood (2013). Yield components of six cultivars of cowpea (Vignaunguiculata (L.) Walp

grown on soil contaminated with spent engine oil.

Spada. (2015). The Role of Vascular Plants Growing on Polymetallic Mineralization Areas in

Immigration of Heavy Metals. mo Oxford: Blackwell Scientific

Surandana (2009) Medicinal plant can be contaminated by this heavy metals via root uptake or

by direct deposition.

Utalangke (2013) Reduces blood pressure and cholesterol and act as an anticancer, antioxidant,

antimicrobial, anti-atherosclerotic.

World Health Organization (WHO, 2007) surveys indicated that about 70-80% of the world

population relies on non-conventional medicine, mainly of herbal sources in their

primary health care.

Zarkada (1997) Moringa leaves are edible and they form parts of tradition diets in many

countries

27