Determination of Heavy Metals in Mentha longifolia of

District Zhob, Balochistan

By
Muhammad Hashim
Program of Study: M.Phil

Supervisor: Dr. Samiullah

Department of Chemistry
University of Balochistan
Session: 2018-2019

i
SYNOPSIS

Determination of Heavy Metals in Mentha longifolia of District Zhob,

Balochistan

Name of candidate: Muhammad Hashim

Father’s Name: Janan

Class: M.Phil

Title/Topic: Determination of Heavy Metals in Mentha longifolia of

District Zhob, Balochistan

Name of institute: Department of Chemistry

University of Balochistan, Quetta.
Supervisor: Dr. Samiullah
Department of Chemistry, University of Balochistan,
Quetta.

ii
 Synopsis
Determination of Heavy Metals in Mentha longifolia of District
Zhob, Balochistan

Submitted for

M.Phil

In
DEPARTMENT OF CHEMISTRY
UNIVERSITY OF BALOCHISTAN, QUETTA

Name of Candidate: Muhammad Hashim

Name of Research Supervisor: Dr. Samiullah, Associate Professor

iii
 Table of Contents

1 Introduction.............................................................................................................1

1.1 Background......................................................................................................2

1.2 Significance.....................................................................................................4

1.3 Objectives........................................................................................................4

2 Literature Review...................................................................................................5

3 Research Methodology...........................................................................................6

5 References...............................................................................................................8

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1 Introduction
The trivial name of Mentha longifolia is wild mint, frequently used as folk
remedy. It is used as superficial medicine in Iran and other countries. Mentha
longifolia acquire various pharmaceutical applications such as; antimicrobial,
gastrointestinal, and nervous system effects. And it also has adverse effects on human
health.
The availability of this plant is an awfully adaptable lower-plants enduring
plant kingdom with peppermint–scented scent. Mentha longifolia have rhizome, with
straight stem 42-120 cm long. And the upper part of plant has 6-12 cm height and 1-3
cm width with greyish-green color. They are found in colonial form having length of
5-5 cm, tall branches and low spikes. The mint plant is found in runners form and
speedy cultivators but show phototropism in great extent.
The chemical compounds found in M. longifolia are mostly oxygenated such
as monoterpenes. Pharmacologically active compound in M. longifolia is Pulegone
followed by other compounds such as; menthone, menthol, isomenthone, 1,8-
cineolepiperitenone and borneol. The oils obtained from M. longifolia have great
pharmaceutical importance and acquire insecticidal, antibacterial, antimicrobial and
antifungal properties. Due to this reason such oils are used in clinical and medical
micro-biology, food preservation, pharmacology and phytopathology (Mikaili, et al.,
2013; Daferera, et al., 2000).
One of the important tasks for nutritionist, ecologist and scientists is the
determination of heavy metals in plants of different area. Contamination of plants by
heavy metals cannot be neglected as plants are important constituents of our diet.
Such food stuffs are rich sources of minerals, fibers and vitamins. Fruits and
vegetables are important sources of antioxidants which are essential for body growth
as well as maintenance of health. Sometimes food materials like fruits are
contaminated by heavy metals due to usage of preservatives, chemical sprays,
industrial waste, fertilizers, mining activities and using wastewater. Food quality is
affected by heavy metals which adversely affect human health (Abbas, et al., 2010;
Akinyele, et al., 2015; Chiroma, et al., 2014; Egharevba, et al., 2017; Mikaili, et al.,
2013; Daferera, et al., 2000; Elbagermi, et al., 2012).

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1.1 Background
Nowadays industrialization and agricultural activities are the main causes of
presence of heavy metals in environment which results in environmental pollution and
also effects human health. Therefore, several studies have been carried out in order to
determine the presence of heavy metals in food and their biological effects on human
health (Olowoyo, et al., 2012).
Elements can be categorized into three groups i.e. major and essential
chemical constituents, micro-elements or water pollutants named as heavy elements
and the extreme case of trace elements. Ca 2+, K+, Na+, Mg+, Cl–, S2– and P3+ are
categorized as major elements and are acquired 30 g/day. On the other hand, the
elements such as Ni, Co, Fe, Zn and I 2 are termed as trace elements and are required
in quantity of <5 g/day. Elements which are required in a minute level, which are also
known as ultra-trace elements, in which Pb, Al, Hg, Ba, Br and Sr and some others
revealed in the literature (Subramanian, et al., 2012).
The important sources of major elements are processed foods, fresh fruits,
herbs and vegetables. These elements are essential for proper growth of our body and
health. However, food products can be contaminated by toxic heavy metals (HMs)
from the process of food industrialization and other harmful environmental activities
(Daferera, et al., 2000).
Metals are mostly non-biodegradable contaminations with long biological
half-lives with a great possibility to be gathered in the different body organs which
results in severe human health effects (Egharevba, et al., 2017).
The mints like plants are also known as therapeutic plants and considered as
contaminated texture for many accepted descriptions and supplements. Due to home
ground medications, it has been considered as high cost of mint. In local area mint is
used as herbal medicine and considered as having no side effects on human health.
Geoclimatic conditions are responsible for the metal’s contents in plants and those
plants having vegetative part acquire high concentration of heavy metals (Abbas, et
al., 2010; Akinyele, et al., 2015; Chiroma, et al., 2014; Egharevba, etal., 2017;
Mikaili, et al., 2013). The closeness of overpowering metals past the sensible limits
can cause metabolic unsettling impacts. For an instance, the lack of nutrients and
abundance of heavy metals like Copper, Iron and Zinc, in basic diet is harmful for
human life. The adverse effect of metals (Cd, Cr, Pb, etc) on human body may be
variable showing different results (Daferera, et al., 2000).
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 For medicinal plants WHO and EPA recommend that heavy metals, Fungal
and Bacterial contamination and pesticide should be checked before using as
pharmaceutical drug. M. longifolia (family: Lamiaceae, Class: Magnoliopsida) is
herbal medicine and used to cure a various disease therefore it is essential to
determine the concentration of heavy metals.
The renal failure, intestinal cancer, bone cancer, hypertension, reproductive
problems may be caused by high concentration of Co, Cu, Cd and Pb in our food such
as vegetables and fruits. The harmful effects on living organisms may be due to
minute exposure to these heavy metals. The abnormal change metabolic activities in
the body and disorder in neurological functions of human brain is also due to these
metals (Farooq, et al., 2008).
The historical background reveals that the most human health problems are
due to such metals. The human health is affected by these metals due to oxidation of
coal, industrialization and global waste of polluted products and methane gas
(Bettinelli, et al., 1996).
WHO and FAO suggested the concentration of heavy metals in fruits and
vegetables like in mint; 0.34, 0.42, and 0.22 mg/kg. Many countries in the world have
developed a regulatory frameworks and guidelines for heavy metals in the food
materials and environment such as Australia and New Zealand (Ismail, et al., 2011).
The modern world is facing a great problem of industrial waste and sewage
water. The agricultural lands which is used for harvesting various edible vegetables,
fruits and crops is highly affected by such wastes. These drainage wastes are favored
due to their rich source of organic matter and nutrients. On the other hand, they also
bear the adverse effects due to presence of metals such as; chromium, manganese,
cobalt, copper, zinc, cadmium and lead as well. The heavy metals absorbed by
vegetables from the polluted air, water and soil lead to enhance contaminations level
in food stuff (Elbagermi, et al., 2012).
The heavy metal compounds or contaminants and nutrients are absorbed and
transferred to different tissues of plants become the part of food chain (Bettinelli, et
al., 1996). The different parts of plants absorb the heavy metals from airborne
deposits, polluted soil and environments contaminated with heavy metals. The sewage
and industrial wastewater are used for irrigation of fruit trees lead to the elevation of
contamination level (Mikaili, et al., 2013).

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1.2 Significance
Plants in the form of Fruits and vegetables are the major part of our diet, are
the feed stock of earth crust and biologicals products. These products are active
ingredients for normal diet of human being. However, the photosynthetic products
also contain the essential and non-essential metals. Our environment contains dust
particles and soil contents which also become part of plant body (Igwegbe, et al.,
2013).
Vegetative part of plants are the major source of fibers, minerals and amine
containing organic compounds which can act as antioxidants for plants secondary
growth. Development of the body and prevent the human beings from various
diseases by maintaining overall health. The fruits quality is affected by the presence
of heavy metals and also causes change in odour and taste as well (Ibraheen, et al.,
2017).
The toxic heavy is documented in this selected manuscript. The metals
concentration is compared with accepted values. The metals which are in range of
comparison are K, Na, Mn, Fe, Co, Cd, Pb and Cu. Safety standards suggested by the
WHO/FAO for copper, cadmium and lead are 40, 0.2, and 0.3 mg respectively per kg
in plants. Sodium, calcium, potassium, manganese, chlorine, phosphorus and sulfur
are needed 50.5g/day. Trace elements zinc selenium, iodine, manganese, chromium,
copper, iron molybdenum, cobalt and nickel are necessary <50.5 g/day (Subramanian,
et al., 2012).
Moreover, this study will determine the effects of heavy metals on human
health. However, water containing more pollutants affects the heavy metals
concentration of agricultural plane area and interferences are found in it may enhance
the concentration of heavy metals (HMs) in plants and their contents which makes the
adverse effect on human health instead of becoming sustainable for health to work
properly.

1.3 Objectives
The significant objectives of this manuscript are listed below:

 Qualitative and quantitative analysis of heavy metals in M. longifolia.

 To compare the heavy metals concentration of the sample with accepted
values revealed in the literature by WHO/FAO.

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  To improve the knowledge about chemical/elemental composition of M.
longifolia.

2 Literature Review
Cultivation may be a ponder having amphoteric character i.e. positive and
negative impacts on our troposphere contents. The foremost source of NEP (National
Economy in Pakistan) is agribusiness. Second priority is given to the selected sector
after national defense. Gross domestic product of Pakistan enhanced by 24.32%. The
complete exports contribute 35 percent to the winning from this division and utilizes
51 percent of include up to labor drive and play outstandingly basic portion in lifting
the life of 70% of rural population. Vegetables are created in tremendous sums other
than the major crops of Pakistan. Vegetables like tomatoes chili, etc. are exchanged to
other countries. In Pakistan economy share of vegetable in send out is around 0.22
percent (Abbas, et al., 2010).
The relative densities of heavy metals are >4.5 g/m3. These metals can be
categorized into 3 categories the first one necessary for biochemical life & 2 nd one has
unknown functional role and the 3rd one is “neutral” for all type of living organisms
including human beings. Accumulation of heavy metals by plants like herbs and
shrubs depends upon the moisture and plant species, pH, temperature, availability of
nutrients, organic compounds and the concentration of heavy metals. The heavy
metals concentration in the soil of agricultural land and water varies from area to area.
The uptake of iron, cadmium and chromium is lower in winters than summer due to
low rate of transpiration although the intake of Cu, Pb, and Ni trapped in winter. The
toxicity of soil is increased due to heavy metals. Those metals with long biological
half-lives are naturally toxic and have great chance of accumulation in different
organs of the body causing undesirable effects (Ibraheen, et al., 2017).
In M. longifolia, different types of toxic heavy metals were investigated. The
sample analyzed by Indians scholars which is taken in the form of selected plant
roots, leaved and stems. The quality of medicinal plants was checked by
quantification of this selected plants (Daferera, et al., 2000).
The atomic absorption spectrometer was used to determine the amount of Pb,
Cu, Ni and Cd in various aliquots of vegetative part of plants in the selected cities of
Libya (Mikaili, et al., 2013).

5
 The secondary waste sources of water are climate and tropical water. The
urban population interest may produce more vegetable and the selected plant growth
(Chiroma, et al., 2014).
The research on heavy metals was quantified in many plants obtained from
various areas. Samples were digested by dry ashing method. AAS were used to
quantify and analyze (Igwegbe, et al., 2013).
Varieties of metals were checked in herbal samples and its constituents by dry
ashing and wet ashing methods (Jena, et al., 2007).

3. Research Methodology
The following steps will be employed for analysis of heavy metal in the
samples of selected plant in the area of Zhob district of Balochistan province.

3.1 Sample Collection

Fresh samples of selected plants will be collected from the local area of
district Zhob of Balochistan province.

i. Washing and Drying

Distilled water will be used to rinse the samples of M. longifolia for the
removal of impurities. The sample collected will be chopped properly to make
uniform pieces which will help to dry the pieces of sample at the equal rate. Then acid
washed clean and labeled porcelain crucibles will be used to place the sample and will
be dried in oven at 150°C for 24 h so that they become stiff and crispy.

ii. Grinding/particle size reduction

The grinder will be used to crush and make fine powder of dried samples. The
grinding process provides smaller particles. Polyethylene bags will be used to store
the dried sample.

iii. Digestion
The method of digestion will be used for the preparation of samples for heavy
metals analysis. For the digestion process mixture of three acids (nitric acid,
perchloric acid and sulfuric acid) will be used and mixture will be heated until the

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 complete digestion of organic matter takes place, leaving behind the mineral oxides in
solution. The time and temperature depend on the type of oxidizing agents of acids
are used (Akinyele, et al., 2015).

iv. Heavy metals analysis

The extracts will be used for determination of Heavy metals by using two
different techniques i.e. Atomic Absorption Spectroscopy and Flame Photometry
(Karpiuk, et al., 2016).

v. Data Collection
Data will be collected and analyzed statistically.

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