Journal of Saudi Chemical Society (2010) 14, 97100

King Saud University

Journal of Saudi Chemical Society

www.ksu.edu.sa
www.sciencedirect.com

ORIGINAL ARTICLE

Determination of major and trace elements in the

indigenous medicinal plant Withania somnifera and
their possible correlation with therapeutic activity
Khaula Shirin *, Saima Imad, Sheraz Shaq, Kaneez Fatima
Pakistan Council of Scientic and Industrial Research, Laboratory Complex, Karachi, Pakistan
Received 21 March 2009; accepted 3 October 2009
Available online 16 December 2009

KEYWORDS
Elements;
Withania somnifera;
Atomic absorption
spectrophotometer;
Therapeutic activity

Abstract Withania somnifera (family Solanaceae) has been studied to determine major and trace
elements and their possible correlation with therapeutic value of the plant. Nine trace elements were
determined in Withania somnifera. This important medicinal plant was found to be rich in Fe, Cu,
Ni, Mn, and Zn. These trace elements are well known for curing diseases. The plant contains nutrient elements, which are best sources for fodder.

1. Introduction
There are many elements present in the living organisms and
scores of metabolic reactions are attributed to them (Suzanne,
2007). Nature has provided man herbal medicines under different conditions and climate. Perhaps all basic principles pertaining to human therapy have been derived from medicinal
plants and herbs (Lai, 2004). It is well established that plants
pick up elements from the soil, e.g., banana takes up potassium
and turmeric accumulates large quantity of lead (Weerasinghe
* Corresponding author.
E-mail address: khaula_ark@yahoo.com (K. Shirin).
1319-6103 2009 King Saud University. All rights reserved. Peerreview under responsibility of King Saud University.
doi:10.1016/j.jscs.2009.12.015

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2009 King Saud University. All rights reserved.

and Premalal, 2002). The elemental composition of many

plants is known. It will be very interesting to determine their
trace elements status. It is surprising to note that many curative effects of medicinal plants used in the traditional system of
medicines are due to the presence of very minute quantities of
trace elements. Important constituents of the body such as enzymes are intimately associated with the chemical elements.
Elements, particularly essential trace elements play both curative and preventive roles in ghting diseases such as Fe in anemia and iodine in goiter. At present about 14 such elements are
considered to inuence the state of health and diseases of animals, plants and human beings. These elements are Fe, Cu, Co,
Ni, Zn, Mg, Mn, Mo, Cr, V, Li, Se, F and I (Seiler et al., 1994).
The deciency of trace elements in human subjects can occur
under most practical dietary conditions. Many diseases which
have been considered incurable may now possibly be treated
by balancing the disequilibrium of these elements in the human
body. Liver diseases such as viral and alcoholic hepatitis do
not easily respond to treatment with drugs of modern allopathic system of medicines (Zhang, 1996). Traditional herbal
system has some thing to offer in this eld. Withania somnifera

98
Dunal (Encyclopedia of Medicinal Plants, 1996) (family Solanaceae) locally known as Ashwagandha is widely distributed
in Pakistan especially in Karachi (Ciddi, 2006). The plant was
selected for elemental analysis mainly due to its availability
and use against a variety of ailments including tumor. The
tuberous roots are astringent, bitter, acrid, somniferous, thermogenic, stimulant, aphrodisiac, diuretic and tonic. They are
useful in vitiated condition of vata, leucoderma, constipation,
insomnia, tissue building and nervous breakdown (Mathur
et al., 2006). The leaves are bitter and are recommended in fever, painful swelling and ophthalmitis (Sudhir et al., 1986).
Several bioactive compounds have been isolated from this
plant like steroidal lactone called Withanolides possessing
antibiotics and antitumor and anxiolytic antidepressant activities (Elsakk et al., 1990; Bhathacharya et al., 2000). Withanolides D occurs in leaves possessing signicant antitumor
activity in vivo against sarcoma 180 and Ehrlich ascites carcinoma and in vitro against cells derived from human epidermoid carcinoma of the nasopharynx (KB) (Jayaparkasam
et al., 2003). This potential plant is a promising drug source
for cancer chemotherapy and radio sensitization (Devi, 1996;
Padmavathi et al., 2006). Withaferin A, an active component
of this medicinal plant has the strong radio sensitization effect
of Ehrlich ascites carcinoma grown in Swiss mice, which offer
great promises in cancer therapy (Uma and Akagi, 1996). The
aqueous extract of the plant exhibits its anti-inammatory
activity against inammatory mediators, i.e., histamine, 5HT and prostaglandins (Aldor et al., 1982). In this way it reduces edema and hypersensitivity of the reactions caused by
certain drugs. Immuno suppressive drugs such as cyclophosphamide, azathioprine, and prednisolone are used to treat
many diseases. But in certain diseases such as ulcerative colitis
(i.e., inammation of colon) these drugs are used but they
cause bone marrow suppression as side effect which is not required. Ashwagandha prevents myelosuppression due to all
three immunosuppressive drugs (Puri, 2002). This miraculous
plant is used in Ayurvedic treatment to treat Parkinsonian patients. It has maculating effects on Cu2+, Zn2+ and Fe3+ levels in serum suggesting that the benecial effect of this
medicinal plant to cure arthritis is by its effect on trace elements metabolism (Kulkarni, 1991).
2. Experimental
Plants were collected from the eld of PCSIR Lab, complex in
the month of June and identied at Botany Department, University of Karachi. The plants were air dried, then dried in
oven at 80 C and powdered. The ash was made in furnace
at 550 C (Martin et al., 1996).
2.1. Digestion of the samples
The ash (1.0 g) was digested with 10.0 ml concentrated hydrochloric acid in acid-washed Pyrex tube till the solution was
cleared and the volume was reduced to about 1.0 ml. The solution was ltered and made up to 10.0 ml with distilled water
(Shirin et al., 2008). Blank was also prepared without adding
the sample. Standard addition technique was used for the estimation of elements. Stock standards of M/s E. Merck were
used for making series of standard addition solutions after
appropriate dilution. The nal acid concentration in all solu-

K. Shirin et al.
tions was 1%. The dilutions of sample solutions were made
such as to keep the concentration of different elements within
the linear range of absorbance.
2.2. Instrumentation
Hitachi Z-5000 Atomic Absorption Spectrophotometer
equipped with Zeeman background corrector and data processor was used for elemental analysis of the samples. All parameters were set and followed strictly according to the
manufacturers instructions using ame atomization technique
(Table 3).
3. Results and discussion
The plant samples were dried at 80 C in order to remove the
combined water which may result in poor homogenization and
incorrect analytical results. The ash was made between 500 and
550 C to prevent the loss of analyte (Shirin et al., 2008) (Table 1). Micro nutrients and toxic elements such as Zn, Cu, Fe,
Cr, Co, Mn, Ni, Pb, and Cd were studied in various parts of
Withania somnifera. These elements were found in signicant
amounts (Table 2). Many therapeutic activities of this plant
may be attributed and correlated to the presence of these potential trace elements. Deciencies of these elements may cause
different diseases.
Zn has been reported to have benecial effects on the atherosclerotic patients as compared to normal (controls). Subnormal plasma Zn level has been reported in patients with
atherosclerosis (Ripa and Ripa, 1994). Several steroidal lactones have been isolated from Withania somnifera and some
of them are reputed for their therapeutic values like Withaferin
A, which possesses both antitumor and antibiotic activities. It
exhibited signicant antitumor activity when tested in vitro
against human carcinoma of nasopharynx [KB] (Bhathacharya
et al., 2000). The presence of Zn and Cu in the plant may be
correlated with its anticancer property, as both elements are required in growth and proliferation of normal cells. Zn concentration decreases in cancer patients whereas Cu concentration
increases (Frans Kok et al., 1988). It is believed that low concentration of plasma Zn in cancer patients is due to the increased requirement of Zn by cancer tissues (Rev. Neurosci.,
1998). This seems to be reasonable because of the fact that tumor cells have high rate of DNA synthesis and most of the enzymes involved in the nucleic acid synthesis are Zn dependent
(Caballero-George et al., 2001). Chronic deciency has been
described in association with dwarsm and hypogonadism in
the Middle East (Food and Nutritional Bulletin, 2004; Vallee
and Falchuk, 1993). Copper deciency in human may result
in hypochromic microcytic anemia, neutropenia and bone
changes (Linder and Hazeg-Azam, 1996). Iron is a structural
component of hemoglobin, myoglobin and other heme pro-

Table 1

Ash of different parts of the plant.

Test sample

ASH (%)

W.S.
W.S.
W.S.
W.S.

85.80
93.73
90.57
94.57

leaves
shoots
seeds
roots

Determination of major and trace elements in the indigenous medicinal plant Withania somnifera
Table 2

The concentration of elements found in Withania somnifera (W.S.).

Test sample

W.S.
W.S.
W.S.
W.S.

99

leaves
shoots
seeds
roots

Trace elements
Zn (ppm)

Cu (ppm)

Fe (ppm)

Cr (ppm)

Co (ppm)

Mn (ppm)

Ni (ppm)

Pb (ppm)

Cd (ppm)

422.2
375
878.8
928.1

245.7
135.8
261.8
306.3

9417.7
3750.2
2380.1
15777.3

11.9
9.0
5.5
70.4

3.2
2.6
N.D.*
16.3

833.5
452.6
421.1
649.6

16.2
10.3
7.3
38.9

60.6
23.3
N.D.
17.6

N.D.
N.D.
N.D.
N.D.

N.D.* > 0.006 ppm.

N.D. > 0.005 ppm.

Table 3

Instrumental conditions for ame atomization.

Condition

Zn

Cu

Fe

Cr

Co

Mn

Ni

Pb

Cd

Lamp current (mA)

Wave length (nm)
Slit width (nm)
Oxidant
Oxidant pressure (kPa)
Fuel
Fuel pressure (L/min)
Burner height (mm)
Calculation mode
Calculation time (s)

6.5
213.9
1.3
Air
160
Air C2H2
2.0
7.5
Integral
5.0

9.0
324.8
1.3
Air
160
Air C2H2
2.2
7.5
Integral
5.0

15.0
248.3
0.2
Air
160
Air C2H2
2.0
7.5
Integral
5.0

9.0
359.3
1.3
Air
160
Air C2H2
2.8
7.5
Integral
5.0

15.0
240.7
0.2
Air
160
Air C2H2
2.2
7.5
Integral
5.0

9.0
279.6
0.4
Air
160
Air C2H2
2.2
7.5
Integral
5.0

12.0
232.0
0.2
Air
160
Air C2H2
2.2
7.5
Integral
5.0

9.0
283.3
1.3
Air
160
Air C2H2
2.2
7.5
Integral
5.0

9.0
228.8
1.3
Air
160
Air C2H2
2.0
5.0
Integral
5.0

teins, many of which are enzymes of the Krebs cycle. Iron deciency also causes anemia (John Beard, 2001). Chromium has
signicant effect in controlling sugar level in the blood of human beings. The Africans and the Orientals have very low
rates of cardiovascular diseases because of the high levels of
chromium in African peoples as much as four times higher
than the North Americans. The high chromium contents in
the tissues of Africans may be due to the use of raw sugar,
which contains an appreciable amount of chromium (Vincent,
2004). Cobalt is a part of vitamin B-12. Its deciency in human
beings seriously affects some biological processes (Sullivan,
2002). Manganese also has antioxidant property and plays a
role in the prevention of toxic oxygen forms. It may play a part
in the degenerative process called aging. It is a catalyst in the
synthesis of fatty acids, cholesterol and mucopolysaccharides
(Dismukes and Van Willigen, 2006). Signicant amounts of
nickel are found in DNA and RNA. Nickel may act as a stabilizer of these nucleic acids (Phipps et al., 2002). Lead and
cadmium have no roles to play in metabolism. Although lead
was found in slight excess cadmium was not detected up to
parts per million level. Excess of lead may be due to absorption
of lead from the environment (Seregin and Ivanov, 2001).
4. Conclusion
It is concluded that studies regarding metals in herbal preparations are useful for mankind. Our study of elements with respect to indigenous medicinal plant reveals that major and
trace elements have signicant roles in combating a variety
of human ailments and diseases. Further research on these elements is yet to be carried out, whose functions are still not
known but are present in humans and other living organisms.

References
Aldor, Y., Walach, N., Modai, D., Horn, Y., 1982. J. Mol. Med. 60
(7), 375377.
Bhathacharya, S.K., Bhattacharya, A., Ghosal, S., 2000. Phytomedicine 7, 463469.
Caballero-George, C., Vanderheyden, P.M.L., Solis, P.N., Proters, L.,
Shahat, A.A., Gupta, M.P., Vanquelin, G., Vlietinck, A.J., 2001.
Phytomedicine 8 (1), 59.
Ciddi, V., 2006. Ind. J. Pharm. Sci. 68 (4), 490492.
Devi, Pu, 1996. India J. Exp. Biol. 34 (10), 927932.
Dismukes, Charles, Van Willigen, T. Rogies, 2006. Encyclopedia of
Inorganic Chemistry, 11151118.
Elsakk, M., Grigorescu, E., Stanescu, U., 1990. Rev. Med. Chir. Soc.
Med. Nat. Lasi. 94, 385387.
Frans Kok, J., Cornelia Vanduijn, M., Hofman, A., Gigsbert, B.,
Voet, V., Fred De Wolf, A., Kok, J., 1988. Epidermiology 128,
352359.
Jayaparkasam, B., Zhang, Y., Seeram, N.P., Nair, M.G., 2003. Life
Sci. 74 (1), 125132.
John Beard, L., 2001. J. Nutr. 131, 568S580S.
Kulkarni, R.R., 1991. J. Ethno-Pharmacol. 33 (12), 9195.
Lai, P.K., 2004. Curr. Med. Chem., 14511460.
Linder, M.C., Hazeg-Azam, M., 1996. Am. J. Clin. Nutr. 63 (5), 797S
811S.
Martin, H.W., Young, T.R., Kaplan, D.I., Simon, L., Adriano, D.C.,
1996. Plant Soil. 182, 199.
Mathur, R., Gupta, S.K., Singh, N., Mathur, S., Kochupillai, V.,
Velpandian, T., 2006. J. Ethnopharmacol. 105 (3), 336341.
Padmavathi, R., Banu, S.M., Sakthisekaran, D., 2006. Chem. Biol.
Interact. 159 (3), 180185.
Phipps, T., Tank, S.L., Wirtz, J., Brewer, L., Coyner, A., Ortego, L.S.,
Fairbrother, A., 2002. Environ. Rev. 10, 209261.
Puri, H.S., 2002. Rasayana. CRC Press, New York, pp. 4650.
Ripa, S., Ripa, R., 1994. Min. Med. 85 (2), 647654.

100
Seiler, H.G., Sigel, A., Sigel, H., 1994. In: Handbook on Metals in
Clinical and Analytical Chemistry, vols. 2021. CRC Press,
Switzerland.
Seregin, I.V., Ivanov, V.B., 2001. Russ. J. Plant Physiol. 48 (4), 523
544.
Shirin, K., Naseem, S., Bashir, E., Imad, S., Shaq, S., 2008. J. Chem.
Soc. Pak. 30 (1), 9096.
Sudhir, S., Budhiraja, R.D., Miqlani, G.P., Arora, B., Gupta, L.C.,
Graq, K.N., 1986. Planta Med. 1, 463469.
Sullivan, K., 2002. Vitamins and Minerals: A Practical Approach to a
Health Diet and Safe Supplementation. Harpers Collins.
Suzanne, S., 2007. In: Elements in Living Organisms, vols. 67. The
Rosen Publishing Group, New York.
Uma, D.P., Akagi, K., 1996. Int. J. Rad. Biol. 69 (2), 193197.
Vallee, B.L., Falchuk, F.H., 1993. Physiol. Rev. 118, 7379.

K. Shirin et al.
Vincent, J.B., 2004. Proc. Nutr. Soc. 63 (1), 4147.
Weerasinghe, P., Premalal, N.H.R., 2002. Ann. Srilanka 4, 109117.
Zhang, M., 1996. Gastroenterology 110 (4), 11501155.

Further reading
Andrew, C., 1996. The Encyclopedia of Medicinal Plants, 303148,
Dorling Kindersley, London.
Christine, H., Kenneth, H.B., 2004. Food and Nutritional Bulletin,
vol. 25, issue 1, The United Nations University, (Suppl. 2).
Dennis, W.C., Jae, Y.K., 1998. Annual Review of Neuro. Science 21
(3), 347375.