The Executive Editor,
Pakistan Journal of Scientific and Industrial Research
Shahrah-e-Dr. Salimuzzaman Siddiqui,
Karachi-75280, Pakistan.
Subject:
Submission of Manuscript.
Dear Sir,
With due respect, it is stated that I submitting a paper in your journal under following
particulars,
Title: Estimation of heavy metals in medicinal plants as a source of herbal medicine used in
cardiovascular diseases.
*
Naeem Abbas, Farah Deeba and Tahira Shafique.
*
Center for environmental Protection Studies,
PCSIR Laboratories Complex Ferozepur Road, Lahore-54600, Pakistan.
Author for correspondence: naeemchemist@gmail.com
I shall be very much obligate, if you would give early response for publishing this paper.
Thanking you,
Yours sincerely,
(Naeem Abbas)
Scientific Officer
CEPS- PCSIR Labs Complex
Ferozpur Road, Lahore –54600
Email: naeemchemist@gmail.com
Cell no:0334-4034282
Estimation of heavy metals in medicinal plants as a source of herbal medicine used in
cardiovascular diseases.
*
Naeem Abbas, Farah Deeba and Tahira Shafique.
*
Center for environmental Protection Studies,
PCSIR Laboratories Complex Ferozepur Road, Lahore-54600, Pakistan.
Abstract
Heavy metal distributions in medicinal plants are gaining importance as an alternative medicine. These
medicinal plants have been used in traditional system for centuries. Tradition systems of medicine
have provided lead over many of the important drug used in modern medicine. The main purpose of
this study is to evaluate the heavy metal concentrations in medicinal plants, which are extensively used
in preparation of herbal medicine for heart disease and tonic for general human health. High Cr
contents were observed in Terminalia Arjuna bark 0.3480 mg/100 gm. The high Mn concentration was
found in Eucalyptus leaf 1.4654 mg/100 gm and Cassia Fistula twig contains high Mg concentration
0.5441 mg/100 gm.
Keywords: Medicinal plant, Terminalia Arjuna, Cassia Fistula, Eucalyptus
Introduction:
Herbal medicine have been using for treatment of different diseases since the origin of human being.
The contribution of medicinal plants in the traditional system for curing diseases has been
documented. Now a day’s development of herbal products as dietary supplements in view of renewed
interest. Oriental herbal medicines have a prominent role to play in the pharmaceutical and health
markets of the 21st century (Kleinschmidt and Johnson, 1977).
The uses of traditional medicinal plants for primary health care have steadily increased worldwide in
recent years. Scientists are in search of new phytochemicals that could be developed as useful antimicrobial for treatment of infectious diseases. Currently, out of 80% of pharmaceuticals derived from
plants, very few are now being used as anti-microbial. Plants are rich in a wide variety of secondary
metabolites that have found anti-microbial properties. (Ramar and Ponnampalam, 2010)
A number of herbal drugs have been mentioned to be affected in various cardiac disorders by ancient
physicians and traditional healers as modern to the investigation and other analytical instruments were
not known in ancient time. Most of the observations about therapeutic efficacy of the drugs were
mainly based on subjective and empirical assessments. Historical data shows that discovery of several
important modern drugs of herbal origin owe to the medical knowledge and clinical expertise of
physicians. (Mohsin Raza, 2006)
Heavy metal analysis of Pb, Cd, Hg, Cu, Ni, Fe, Mn, Cr and As in medicinal plants is getting lot of
scientific interest as these plants are used as an alternate form of medicine in different parts of the
world.( Samy and Gopalakrishnakone, 2008: Kala, 2005: Katewa et al., 2004: Wong et al., 1993:
Abdul-Wahab et al., 2008). In food heavy metals are present in very minute quantities. It has been
established that whatever is taken as food might cause metabolic disturbance if it does not contain the
permissible upper and lower limits of heavy metals. Thus, both deficiency and excess of essential
micro nutrients (eg, Zinc, Cd, Ni, Cr etc) may produce undesirable effects (Konofal et al., 2004:
Kocak et al., 2005).
Biological function of trace elements:
Chromium:
The beneficial effects of chromium are cardiovascular diseases were first hypothesize by Schroeder
and coworkers after their observation. There is evidence that chromium has a role in the pathogenesis
of cardiovascular disease in experimental animals. Chromium deficiency leads to elevation of serum
cholesterol. Regression of cholesterol induced aortic plaques during chromium feedings to rabbits has
been described. It has been suggested that chromium influences the number are affinity of insulin
receptors in peripheral tissues in agreement this hypothesis chromium supplementation has in some
studies lowered the response to plasma insulin to glucose loads.(Uusitupa et al., 1983) These finds are
important as hyperinsulinemia is possibly an important risk factor for coronary disease in man.
Manganese:
Magnesium acts as cofactor of many enzymes including mitochondrial superoxide dismutase and
pyruvate carboxylase. (Hurley and Keen, 1987.) Manganese is necessary for thiamin (Vit.B1) and
Vit.E metabolism, therefore a deficiency in manganese can compromise nutritional status with respect
to vitamins.
Magnesium:
It has been reported that magnesium deficiency may enhance aging via protein oxidation. ( Gunter and
Hollriegl, 1994) Magnesium deficiency induces cardiac arrhythmias both in experimental animals and
humans. A transient hypomagnesaemia is a common finding in patients with acute myocardial
infarction.
There
is
evidence
that
administration
of
intravenous
magnesium
prevents
hypomanganesemia, diminishes size of myocardial infarction and results in lower mortality rates in
patients with acute myocardial infarction. (Roden, 1989)According to US National Academy of
Science (1977), there have been more than 50 studies in nine countries that have indicated an inverse
relationship between water hardness and mortality from cardiovascular disease. That is why people
who drink water that is deficient in magnesium and calcium generally appear more susceptible to this
disease.
Copper
Copper has been shown to be important component of many protein and enzymes. Sever copper
deficiency has been associated with cardiac abnormalities in several animal species. Aortic rupture in
avian species, bone abnormalities and anemia in some species are reported as clinical features of
copper deficiency. (Das, 1990)
Zinc
Zinc has been shown to be necessary for the function of many enzymes. Low zinc status in guinea pigs
has been found to impair calcium uptake by brain synaptosomy.(Browning, 1994) Zinc, copper,
magnesium and potassium have been shown as depressor of hypertension.(Arora et al., 1986)
Symptoms of mild deficiency are diverse as the mineral plays so many different functions.
Material and Methods:
Collection and Post Harvest Treatment of Plant Material
Experiment was carried out at PCSIR laboratories Lahore. Plants were collected from natural habitat
of PCSIR vicinity. Each type of medicinal plant species was collected from 10 locations from each
sampling site. Result reported was arranged of all 10 samples within an area of approximately one
square kilometer to get better sampling representation. Samples were washed in fresh running water to
eliminate dust, dirt and possible parasite, and then treated with deionized water.
Quality Control
Each demoisturized plants sample was crushed in an agate mortar as fine as possible. Each sample was
analyzed three times.
Acid Digestion of Plant Samples
Weighed quantities of crushed and powdered portion from each part of plant; leaf, bark, fruit and twig
in a crucible were heated in an oven at 110oC to remove moisture. The dried sample were charred and
then heated in a furnace for 4h at 550oC. The contents of crucible were cooled in desiccators and 5 mL
concentrated HNO3 was added into the dish to dissolve its contents. The solution was filtered and
transferred to a 100mL flask and diluted up-to the mark (Radojevic, 1999). Estimation of heavy metals
was carried out on atomic absorption spectrometer, analyst 800 (Perkin Elmer).
Reagents
All reagents were of analytical reagent grade and high purity distilled water was used for making the
solutions.
Result and discussion
As mention earlier that Cr, Zn, Cu, Mn and Mg are some element that involved in antioxidant defense
mechanism. The functions of heart, both in pathological and physiological state are affected by these
elements in verity of ways. Though exact mechanism is not clear, it has been reported that at least 31
elements can be linked to cardio vascular disorder. (Vohrora, 1983).
There is rather strange that a thousand years ago when there was no developed technology. The
physicians identified a large no of herbs which later on proved to medically useful in cardiac ailments.
We have selected three plants Eucalyptus, Cassia Fistula and Terminila Arjuna for studying because of
their medicinal values. The concentrations of Zn, Cu, Mg, Mn and Cr in selected medicinal plants are
appended (Table 1).
Botanical Name: Terminalia Arjuna
Part used: Fruit, leaf, bark, twig
Botanical description
Locally known as kumbuck belong to the family of combretaceae (Sarveswaran et al., 2006). Arjuna is
a large deciduous tree with a height of about 60 to 80 feet and trunk 10 to 12 feet circumference. It has
a leave 3 to 8 inches broadly elliptic, cuneate at base and clustered at the ends of branchlets. ( Kapoer,
2005)
Therapeutic Uses
Arjuna is popularly used as cardiac tone. It is very useful in diarrhea, dysentery and spure. Its fruit is
useful in cough, hoarseness and eye diseases. Arjuna bark provides positive support to the
cardiovascular system. It
help with the prevention of atherosclerosis, lowering blood pressure,
reducing angina attacks, increasing capillary integrity, reducing edema, preventing destructive enzyme
activity, and lowering LDL cholesterol and triglyceride levels. (Johanne et al., 1999)
Elemental composition
Elemental composition of Terminalia Arjuna shows that its fruit is very rich in Cu (18.248 mg/100
gm), Mg (0.417 mg/100 gm) shown in Table 1.Leterature shows that its bark is recommended in heart
diseases, it contain maximum Cr as compared to other selected medicinal plant shown in (Fig 1). The
Mg concentration in its bark is (0.9602 mg/ 100 gm). The zinc concentration in fruit of Terminalia
Arjuna is very high as compared to eucalyptus and cassia fistula was shown in (Fig 4). The dietary
limit of Zn is 100 mg/L. (Jones, 1987).
Botanical Name: Cassia fistula
Part used: Fruit, leaf, bark, twig
Botanical description
Cassia is a small to medium sized tree with compound leaves and large shining, dark green leaflets. It
has hanging branches and black or shining dark brown, 50 to 60 cm long cylindrical fruit. Its flowers
are bright yellow in colour, drooping racemes, 30-60 cm long; shortly clawed petals, to 3.5 cm across;
stamens 10, upper three with erect filaments to 0.7 cm long. (Orwa et al. 2009)
Therapeutic
Uses
The leaves of the cassia tree are helpful in relieving irritation of the skin and in alleviating swellings
and pains. The cassia pulp is a popular laxative and is used in the treatment of constipation. Its bark
extracts possess significant anti-inflammatory and anti oxidant properties. (Raju et al., 2005).The root
of the cassia tree is a tonic and useful in reducing fever.
Elemental composition
Elemental analysis of Cassia Fistula shows that Zn and Cu are not detectable in all part of plant.
However, Mg concentration was remarkable shown in (Fig 2). Its twig is very rich in Cr (0.1248
mg/100 gm) and very low amount of Mn (0.0035 mg/100 gm). Its fruit contain maximum
concentration of Mn shown in (Fig 3).
Botanical Name: Eucalyptus
Part used: Fruit, leaf, bark, twig
Botanical description
Eucalyptus is a diverse genus of flowering trees (and a few shrubs) in the myrtle family and having
subfamily Myrtaceae. Nearly all Eucalyptus are evergreen but some tropical species lose their leaves
at the end of the dry season. Eucalyptus leaves are covered with oil glands. The copious oils produced
are an important feature of the genus. Flowers have numerous fluffy stamens which may be white,
cream, yellow, pink or red; in bud, the stamens are enclosed in a cap known as an operculum which is
composed of the fused sepals or petals or both. Thus flowers have no petals, but instead decorate
themselves with the many showy stamens.
Therapeutic Uses
It is a fast-growing source of wood, its oil can be used for cleaning and functions as a
natural insecticide, and it is sometimes used to drain swamps and thereby reduce the risk of malaria.
Outside their natural ranges, eucalypts are both lauded for their beneficial economic impact on poor
populations and derided for being invasive water-suckers, leading to controversy over their total
impact.
Elemental composition
Elemental analysis of Eucalyptus shows that its fruit is very rich in Mg (1.4772 mg/100 gm), Zn
(0.1872 mg/100 gm) and Cr (0.0985 mg/100 gm). Their leaves contain maximum Mn as compared to
Terminalia Arjuna and Cassia Fistula shown in (Fig 3). Mg content in its leaf, bark and twig are
remarkably shown in (Fig 2).
Conclusion
These medicinal plants have been used since a thousand year ago and their metals content in addition
to their essential oils may contribute towards their effectiveness. The present study shows the presence
of essential metals in different parts of plant body. The finding may be utilized in synthesis of herbal
medicines for heart, liver diseases and tonic for general human health.
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Table 1: Heavy metal concentration (ppm) in plant materials on dry basis as mg/100 gm
Plant species
Terminalia
Arjuna
Eucalyptus
Cassia Fistula
Plant part
Zn
Cu
Mg
Mn
Cr
mg/100 gm
mg/100 gm
mg/100 gm
mg/100 gm
mg/100 gm
Fruit
2.2019
18.248
0.417
0.0746
0.0246
Leaf
N.D
N.D
0.6171
0.0091
0.1594
Bark
N.D
N.D
0.9602
0.0377
0.3480
twig
0.2543
0.1696
1.0164
0.3931
0.1304
Fruit
0.1872
N.D
1.4772
0.4169
0.0985
leaf
0.3993
0.1962
1.2852
1.4654
0.1212
Bark
.00617
0.0456
1.055
0.2216
0.0406
twig
0.0828
0.2123
0.7766
0.3241
0.0248
Fruit
N.D
N.D
0.3480
0.0061
0.0193
Leaf
N.D
N.D
0.2729
N.D
0.1075
Bark
N.D
N.D
0.1922
0.0102
0.0941
twig
N.D
N.D
0.5441
0.0035
0.1248
N.D= Non detectable
Cr conc.(mg/kg)
0.4
Total Cr
0.3
0.2
0.1
T.
F
ru
T. it
Le
a
T. f
Ba
r
T. k
Tw
E. ig
Fr
ui
t
E.
lea
E. f
Ba
r
E. k
Tw
C. ig
Fr
ui
C. t
lea
C. f
Ba
r
C. k
.T
wi
g
0
Plant part used
Fig.1: The amount of total Chromium in some selected medicinal plant.
T.
F
ru
T. it
Le
T. a f
Ba
r
T. k
Tw
E. ig
Fr
ui
t
E.
le
a
E. f
Ba
r
E. k
Tw
C ig
.F
ru
i
C t
.le
a
C f
.B
ar
C k
..T
w
ig
Mg. conc.(mg/kg)
Total Mg
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
Plant part use d
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
Total Mn
T.
F
ru
T. it
Le
T. a f
Ba
r
T. k
Tw
E. ig
Fr
ui
E. t
le
E. a f
Ba
r
E. k
Tw
C ig
.F
ru
C it
.le
C af
.B
a
C rk
..T
w
ig
Mn conc. (mg/kg)
Fig.2: The amount of total Magnesium in some selected medicinal plant.
Plant part use d
Fig.3: The amount of total Manganese in some selected medicinal plant.
ru
T. it
Le
T. a f
Ba
r
T. k
Tw
E. ig
Fr
ui
t
E.
le
a
E. f
Ba
r
E. k
Tw
C ig
.F
ru
i
C t
.le
a
C f
.B
ar
C k
..T
w
ig
T.
F
Zn conc.(mg/kg)
2.5
2
Total Zn
1.5
1
0.5
0
Plant part use d
Fig.4: The amount of total Zinc in some selected medicinal plant.