R.R.sheeja and Beena Lawrence

Int.J.Curr.Microbiol.App.
Sci (2015) 4(3): 344-347
ISSN: 2319-7706 Volume 4 Number 3 (2015) pp. 344-347

http://www.ijcmas.com
Original Research Article

Phytochemical Screening of the Leaves of Stevia rebaudiana, Bertoni
R.R.Sheeja1* and Beena Lawrence2

1
Bethlahem College of Education, Karungal, Kanyakumari District, Tamil Nadu, India
2
Department of Botany, Women s Christian College, Nagercoil, Kanyakumari District,
Tamil Nadu, India
*Corresponding author
ABSTRACT
Keywords Nature has store house of remedies to cure all ailments of mankind. From the
ancient ages the plants have been used for medicinal uses and other useful proposes
Bioactive to humans. The present investigation was carried out to estimate the phytochemical
compounds, constituents present in Stevia rebaudiana Bertoni. The results showed the presence
Stevia of bioactive constituents of alkaloids, tannins, flavonoids, glycosides, saponins,
rebaudiana, quinone and triterpenes. Catechins, coumarins and xanthoproteins were not
alkaloids, detected. More research work is recommended on the plant leaves for isolation and
tannins. characterization of bioactive compounds.
Introduction
Materials and Methods
Stevia rebaudiana Bertoni is a versatile herb
with incredible sweetness that is gaining Collection of plant materials:
very high popularity amongst all type of
sweetener users as most ideal substitute for Stevia rebaudiana plant were obtained from
sugar. It produces sweet steviol glycosides. Thiruvananthapuram, Kerala and was grown
It is a high demanding antidiabetic under protective conditions. The flora of
medicinal plant belonging to Asteraceae presidency of Madras (Gamble,1935) and
family. the Flora of Tamil Nadu Carnatic
(Matthew,1983) were used for identification
It is perennial and endemic, medicinal herb and authentication of the plants. The leaves
(Sivaram and Mugundan, 2003). It is also are collected and washed thoroughly in
called as honey plant due to its sweetness. running tap water and rinsed in distilled
Thus the present study is a preliminary water and dried under shade. After drying,
attempt to identify some of the the leaves were powdered using mixer
phytochemicals of the selected plant. grinder and then kept in well closed
Hopefully this will lead to new information container.
on this plant application and new
perspective on the potential use of Stevia.
344
Int.J.Curr.Microbiol.App.Sci (2015) 4(3): 344-347
Preparation of phytochemical extracts Test for triterpenes: Ten milliliter aqueous

extract was placed in a small beaker and
This powder was extracted in the soxhlet evaporated to dryness. The residue was
using ethanol and ethyl acetate and dissolved in 0.5 ml each of acetic anhydrine
subjected to qualitative phytochemical and chloroform.
screening for the identification of various
chemical constituents using the method The solution was transferred into a dry test
described by Trease and Evans (1987) and tube and concentrated sulphuric acid was
by Harbone (1973). The plant extract were added. Brownish red or violet rings at the
screened for the presence of secondary zone of the contact with the supernatant and
metabolites such as alkaloids, flavonoids, green or violet coloration denoted the
tannins, saponins, triterpenes, glycosides, presence of sterols and triterpenes
catechin, coumarin, quinone and
xanthoprotein. Test for Flavonoid (Shindo s test): To the
test solution, a few magnesium turnings and
Test for tannins: About 0.5g of the dried a few drops of concentrated hydrochloric
power was boiled in 20 ml of water in a test acid were added and boiled for five minutes.
tube and then filtered. A few drops of 0.1% Appearance of red or orange red colour
ferric chloride was added and was observed indicates the presence of flavonoids.
for brownish green or a blue black
coloration. Test for Catechin
Test for alkaloids: One milliliter of aqueous To the test solution, a few drops of Echrlich
extract was stirred and placed in 1% reagent and concentrated hydrochloric acid
aqueous hydrochloric acid on a stream bath. were added. Appearance of pink colour
Then, 1 ml of the filtrate was treated with indicates the presence of catechin.
Dragendorff s and Mayer s reagent.
Turbidity or precipitation with this reagent Test for Coumarin
was considered as evidence for the presence
of alkaloids. To 2 ml of the test solution, a few drops of
alcoholic sodium hydroxide were added.
Test for glycosides: 0.5g extract of sample Appearance of yellow colour indicates the
was dissolved in 1 ml water and then presence of coumarin.
aqueous sodium hydroxide was added.
Formation of yellow color indicated the Test for Quinone
presence of glycosides.
The test solution was treated with a few
Test for saponins: 0.5 g extract were drops of concentrated sulphuric acid or
dissolved in 10ml of distilled water in a test aqueous sodium hydroxide solution. Colour
tube was stopperred with a cork and shaken formation indicates the presence of quinone
vigorously for about 30 seconds. The test compound.
tube was allowed to stand in a vertical
position and observed over a 30 minutes Test for Xanthoprotein
period of time. If a honey comb froth above
the surface of liquid persists after 30 To the test solution, a few drops of
minutes the sample is suspected to contain concentrated nitric acid and few ml of
saponin. ammonia were added. Appearance of a red
345
precipitate indicates the presence of coumarins, saponins and xanthoproteins

xanthoprotein. showed negative result. The presence of
these secondary metabolites suggests that
Result and Discussion the plant might be of industrial and
medicinal importance. Several reports say
The preliminary phytochemical analysis of that the compounds possess remarkable
the leaf extract revealed the presence of antitumor, antidiabetic and antioxidant
alkaloids, tannins flavonoids, glycosides, activity (Gupta and Sharma, 2006; Kaur and
saponins, quinone and triterpenes (Table 1 Kaopoor, 2002 and Ray and Hussan, 2002).
and 2). The most abundant compounds in
the ethanol leaf extract were the glycosides. These results give a picture that the plant has
Alkaloids and tannins, were also seen in quite a number of chemical constituents,
higher amounts but lesser than glycosides. which may be responsible for the many
Flavonoids and glycosides were seen in pharmacological actions. They were known
moderate levels, and triterpenes and to show medicinal activity as well as
saponins were seen in least amounts. The exhibiting physiological activity (Sofowara,
test for catechins, coumarins, quinones and 1993). The presence of these
xanthoproteins showed negative result. phytochemicals in the investigated
medicinal plant would be responsible for the
In the ethyl acetate leaf extract the most antimicrobial activity of the plant too. This
abundant compounds were the glycosides finding supports the traditional knowledge
and tannins. Alkaloids and flavonoids, were in selecting the most active medicinal plants
also seen in higher amounts but lesser than to use in traditional medicine practices in the
glycosides. Flavonoids and triterpenoids future. Further work is needed to isolate
were seen in moderate levels, and quinine is active principle from the plant and to carry
seen in least amount. The test for catechins, out pharmaceutical studies.
Table.1 Phytochemical constituents of Stevia rebaudiana leaf with ethanol extract
Phytochemical Results*
constituents
Alkaloids +++
Flavonoids ++
Tannins +++
Glycosides +++
Catechin -
Coumarins -
Saponins +
Quinone -
Triterpenes ++
Xanthoproteins -
346
Table.2 Phytochemical constituents of Stevia rebaudiana leaf with ethyl acetate extract
Phytochemical Results*
constituents
Alkaloids ++
Flavonoids ++
Tannins +++
Glycosides +++
Catechin -
Coumarins -
Saponins -
Quinone +
Triterpenes ++
Xanthoproteins -
+++ Strongly present; ++ Present;
+ Weakly present; - Absent
References Spectrum Books Ltd., Ibadan,

Nigeria, 289 Pp.
Gamble, J.S. 1935. Flora of the presidency Trease, G.E., Evans, W.C. 1987.
of Madras. Adland and Son s Ltd, Pharmacognsy.13th edn. Brailliar
London, U.K. Tiridel Can. Macmillian Publishers.
Gupta, V.K., Sharma, S.K. 2006. Plants as
natural antioxidants. Nat. Prod. Rad,
2006: 326 34.
Harbone, J.B. 1973. Phytochemical
methods. London. Chapman and
Hall, Ltd, Pp. 49 188.
Kaur, C., Kapoor, H.C. 2002. Antioxidant
activity and total phenolic content of
some asian vegetables. Int. J. Food.
Sci. Tech., 37: 153 61.
Matthew, K.M. 1983. The flora of Tamil
Nadu Carnatic. In the Rapinat
Herbarium. St.Joseph s College,
Thichirappalli, India.
Ray, G., Hussan, S.A. 2002. Oxidant,
antioxidant and carcinogenesis.
Indian J.Exp. Biol., 40: 1213 32.
Sivaram, L., Mukundan, U. 2003. In vitro
culture studies on Stevia rebaudiana.
In vitro Cell Dev.Biol., 39(5): 520
523.
Sofowara, A. 1993. Medicinal plants and
traditional medicine in Africa.
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Int.J.Curr.Microbiol.App.

Sci (2015) 4(3): 344-347

ISSN: 2319-7706 Volume 4 Number 3 (2015) pp. 344-347

Original Research Article

R.R.Sheeja1* and Beena Lawrence2

Preparation of phytochemical extracts Test for triterpenes: Ten milliliter aqueous

precipitate indicates the presence of coumarins, saponins and xanthoproteins

Table.1 Phytochemical constituents of Stevia rebaudiana leaf with ethanol extract

References Spectrum Books Ltd., Ibadan,

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