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PHYTOCHEMICAL ANALYSIS OF CLITORIA TERNATEA LINN., A VALUABLE

MEDICINAL PLANT

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J. Indian bot. Soc. Vol. 92 (3&4) 2013 : 173-178

PHYTOCHEMICAL ANALYSIS OF CLITORIA

TERNATEA LINN., A VALUABLE MEDICINAL PLANT
P. MANJULA, CH. MOHAN, D. SREEKANTH, B. KEERTHI AND
B. PRATHIBHA DEVI
Biotechnology and Molecular Genetics Laboratory,
Department of Botany, Osmania University,
Hyderabad-500007, India.
The present paper deals with phytochemical studies in Clitoria ternatea Linn.. It is commonly known as 'butterfly pea'
and “shankhapushpi”. It is a traditional Ayurvedic medicinal plant belonging to the family Fabaceae. The plant extracts
were subjected to phytochemical analysis for screening of medicinal constituents. Valuable data has been collected
pertaining to the presence of various phytochemicals like Alkaloids, Tannins, Glycosides, Resins, Steroids, Saponins,
Flavonoids and Phenols. Further, quantitative estimation of total Flavonoids, Saponins and Phenols was also carried out
which has provided information regarding the medicinal potential of the plant.

Key words: Clitoria ternatea, phytochemical analysis, qualitative analysis, quantitative analysis.

Clitoria ternatea Linn. is an attractive perennial as possessing anxiolytic, antidepressant,

climber with conspicuous blue or white anticonvulsant, antistress (Jain et al. 2003),
flowers. It belongs to the family Fabaceae and sedative (Kulkarni et al. 1988), antipyretic, anti-
commonly known as “butterfly pea” and inflammatory, analgesic (Devi et al. 2003,
“shankhapuspi”. It is traditionally used to treat Gomez and Kalamani 2003), Anthelmintic
various ailments (Sivarajan and Balachandran (Salhan et al. 2011) and anti-microbial activities
1994, Kokate 1999). The plant is native to (Kamilla et al. 2009). The extract of C. ternatea
south-east Asia and distributed in tropical Asia has been shown to improve learning ability,
including India, the Philippines and enhance memory, increase apical and basal
Madagascar (Anonymous 1998). Roots, seeds dendritic branches, and increase acetylcholine
and leaves of C. ternatea are commonly used in content and acetyl cholinesterase activity in rats
the Ayurvedic system of medicine. Extracts of (Rai et al. 2001). The plant contains several
this plant have been used as an ingredient in the secondary metabolites such as kaempferol and
Ay u r v e d i c ' M e d h y a R a s a y a n a ' a s a its glucoside–clitorin, taraxerol and a lactone
rejuvenating recipe used for treatment of aparajitin (Barik et al. 2007). Seeds contain -
neurological disorders and are considered to Sistosterol, hexacosanal, and anthoxanthin
enhance the intellect (Sharma and Dash 1988). (Yoganarasimhan 2000).
The whole plant and seed extracts are used for Phytochemical screening of medicinal plants is
stomatitis, piles, sterility in females, very important in identifying new sources of
hematemesis, insomnia, epilepsy, psychosis, therapeutical and industrial importance (Salhan
leucorrhea and polyurea (Yoganarasimhan et al. 2011). Phytochemical analysis of methanol
2000). The roots are bitter, refrigerant, laxative, extract of Clitoria ternatea roots confirmed the
intellect-promoting, diuretic, anthelmintic, presence of tannins and resins and certain other
tonic and are useful in dementia, hemicrania, constituents (Terahara et al. 1996, Uma 2009,
burning sensations, leprosy, inflammation, Manalisha and Chandra 2011). The present study
leucoderma, bronchitis, asthma, pulmonary deals with the phytochemical analysis of
tuberculosis, ascites, fever, otalgia, different plant parts of Clitoria ternatea for the
hepatopathy and as a cathartic (Nadkarni 1976). presence of Alkaloids, Tannins, Glycosides,
The root, stem and flower are also used for the Resins, Steroids, Saponins, Flavonoids and
treatment of snake bite and scorpion sting Phenols. Quantitative analysis of root extract for
(Morris 1999). C. ternatea has been shown to total Flavonoids, Saponins and Phenols and
have number of pharmacological activities such shoot, flower and seed extract for total
flavonoids was also carried out.
Received on May 08, 2013 Accepted on May 21, 2013
P. MANJULA, CH. MOHAN, D. SREEKANTH, B. KEERTHI AND B. PRATHIBHA DEVI 174

Table 1. Qualitative analysis of the plant extracts of Clitoria ternatea to screen

for the presence of phytochemicals.

+ Presence of the compound.

- Absence of the compound.
Table 2. Quantitative analysis of the aqueous extracts of Clitoria ternatea for
estimation of phytochemicals

* The value is the average of studies conducted in triplicate.

PHYTOCHEMICAL ANALYSIS OF CLITORIA TERNATEA LINN. 175

Figure: 1. (a – b): a. Clitoria ternatea with blue flower b. Clitoria ternatea with white flower.

MATERIALS AND METHODS Test for Tannins:

Clitoria ternatea Linn. plants were collected Five grams of the ground powder was extracted
from Botanical garden, Department of Botany, with 10 ml ammonical chloroform and 5 ml
Osmania University, Hyderabad. The plant chloroform. The mixture was filtered and the
parts namely leaves, roots, shoots, flowers and filtrate was shaken with 10 drops of 0.5 M
seeds were shade dried and powdered in a sulphuric acid. Creamish white precipitate was
mechanical grinder for preparation of extract. observed for the presence of tannins.

Preparation of plant extracts Test for Glycosides:

The powdered plant parts were Soxhlet- About 0.5 gm of methanol extract was taken in a
extracted with methanol. The extract, on test tube and 1 ml glacial acetic acid containing
removal of solvent in vacuum, gave a dark traces of ferric chloride was added to it. To this
greenish brown semisolid residue. The solution, 1 ml concentrated sulphuric acid was
powdered material or the extracts of the plant added and observed for the formation of reddish
parts mentioned above were used for the study. brown colour at the junction of the two layers
and the upper layer turned bluish green in the
Qualitative analysis presence of glycosides.
It comprised of tests for the presence of
Alkaloids, Tannins, Glycosides, Resins, Test for Resins:
Steroids, Saponins, Flavonoids and Phenols. For the tests concerning the presence of Resins,
0.5 gm of methanol extract was taken in a test
Test for Alkaloids tube and 5 ml of distilled water was added to it
About 0.5 gm of methanol extract was taken in and observed for turbidity which indicates the
a test tube and was diluted and homogenized presence of Resins.
with 10 ml distilled water, dissolved in 20 ml
dilute HCl solution and clarified by filtration. Test for Steroids:
The filtrate was tested with Drangendroff's and About 0.5 gm of methanol extract was taken in a
Mayer's reagent. The treated solution was test tube and 2 ml of acetic anhydride was added
observed for precipitation of white or creamy to it and 2 ml of sulphuric acid was added by the
colour. sides of the test tube and observed for the colour
change to violet or blue green.
P. MANJULA, CH. MOHAN, D. SREEKANTH, B. KEERTHI AND B. PRATHIBHA DEVI 176

Test for Saponins: extracted compound).

About 0.5 gm of methanol extract was taken in
a test tube and 5 ml distilled water was added to Determination of Saponins:
it. The solution was shaken vigorously and The method of Obadoni and Ochuko (2001)
observed for persistent froth. The frothing was was used for determination of Saponins. The
mixed with 3 drops of olive oil and shaken root extract (20 gm) was put into a conical flask
vigorously after which it was observed for the and 100 ml of 20 % aqueous ethanol was added.
formation of an emulsion. It was heated over a hot water bath for 4 h with
continuous stirring at about 55º C. The mixture
Test for Flavonoids: was filtered and the residue re-extracted with
About 0.5 gm of extract was introduced into 10 another 200 ml 20 % ethanol. The combined
ml of ethyl acetate in a test tube and heated in extracts were reduced to 40 ml over water bath
boiling water for 1 min. The mixture was then at about 90º C. The concentrate was transferred
filtered. About 4 ml of the filtrate was shaken into a 250 ml separator funnel and 20 ml of
with 1 ml 1% aluminium chloride solution and diethyl ether was added and shaken vigorously.
incubated for 10 min. Formation of yellow The aqueous layer was recovered while the
colour in the presence of 1 ml dilute ammonia ether layer was discarded. The purification
solution indicated the presence of flavonoids. process was repeated and 60 ml of n-butanol
was added. The n-butanol extract was washed
Test for Phenols: twice with 10 ml of 5 % aqueous sodium
About 0.5 gm of extract was taken in a test tube, chloride. The remaining solution was heated in
mixed with 100ml distilled water and heated a water bath. After evaporation, the samples
gently. To this, 2 ml of ferric chloride solution were dried in the oven to a constant weight. The
was added and observed for the formation of content of Saponins was estimated as mg/gm of
green or blue colour. extracted compound.

Quantitative analysis Determination of Phenols

Quantitative analysis of the root extract was The method Gupta et al. (2010) was followed
carried out for total Flavonoids, Saponins and presently. To 5 gm of the root extract in a 250 ml
Phenols and the shoot, flower and seed extract beaker, 200 ml of 10 % acetic acid in ethanol
for total flavonoids. The root extract was was added, covered and allowed to stand for 4
prepared as explained above. h. This was filtered and the extract was
concentrated on a water bath to one quarter of
Determination of total Flavonoids: the original volume. Concentrated ammonium
The Aluminium chloride colorimetric method hydroxide was added drop wise to the extract
(Chang et al. 2002) with some modifications until the precipitation was complete. The whole
was used to determine total Flavonoids content. solution was allowed to settle and the
The liquid extract was prepared (with mixing precipitate was collected and washed with
0.5 gm of root/shoot/flower/seed extract in 100 dilute ammonium hydroxide and then filtered.
ml of water) and 1.0 ml of this was mixed with The residue comprising of the phenols was
1.0 ml of methanol, 0.5 ml of aluminum dried, weighed and expressed as mg/gm of
chloride (1.2 %) and 0.5 ml of potassium extracted compound..
acetate (0.1176 %). The mixture was allowed to
stand for 30 min at room temperature. Later, the RESULTS AND DISCUSSION
absorbance was measured at 415 nm in a Phytochemical screening of medicinal plants is
spectrophotometer. Quercetin was used as very important in identifying new sources of
standard. Flavonoid content is expressed in therapeutical and industrial importance (Salhan
terms of quercetin equivalent (mg/g of et al. 2011). The present study contributes
PHYTOCHEMICAL ANALYSIS OF CLITORIA TERNATEA LINN. 177

valuable information of bioactive compounds were reported in root extract by Manalisha and
present in Clitoria ternatea. Qualitative Chandra (2011) and in shoot and flower extract
analysis of plant extract was carried out for by Uma (2009) and seed extract by Kamilla et
Alkaloids, Tannins, Glycosides, Resins, al. (2009). Flavonoids have been reported to
Steroids, Saponins, Flavonoids and Phenols. possess many useful properties, including anti-
F u r t h e r, s o m e p l a n t e x t r a c t s w e r e inflammatory, oestrogenic, enzyme inhibition,
quantitatively analyzed for Saponins, antimicrobial, anti-allergic, antioxidant,
Flavonoids and Phenols. vascular and cytotoxic anti-tumour activity
Alkaloids are produced by large variety of (Harbone and Williams 2000). Presentiy,
organisms including bacteria, fungi, plants and Phenols were reported in the root extracts of
animals and some alkaloids have a bitter taste Clitoria ternatea and agrees with Gupta et al.
while many are toxic to other organisms (Gupta (2010). Primarily, phenolic compounds are of
et al. 2010). In the present study, alkaloids are great importance as cellular support material
present in shoot, flower and seed extracts, because they form the integral part of cell wall
which agrees with Uma (2009) and Chauhan et structure by polymeric phenolics (Gupta et al
al. (2012) and absent in leaves and shoot 2010). Bioactive polyphenols have attracted
extracts (Rao and Savitramma 2011). Tannins special attention because they protect the
and Resins reported in the present study in human body from the oxidative stress which
shoots, leaves, flowers and seeds agrees with may cause many diseases, including cancer,
the findings of Rao and Savitramma (2011), cardiovascular problems and ageing (Robards
Uma (2009), Manalisha and Chandra (2011). It et al. 1999).
was attributed that tannins contributed the The root extracts were quantitatively analyzed
property of astringency leading to faster for secondary metabolites like Flavonoids,
healing of wounds and inflamed mucous Saponins, and Phenols. The shoot, flower and
membranes (Okwu and Josiah 2006). Whereas seed extracts were quantitatively analyzed for
Terahara et al. (1996), Uma (2009) and total Flavonoids (Table-2). A high percentage of
Manalisha, and Chandra (2011) reported the Flavonoids were observed in the aqueous
presence of Tannins and Resins in the roots, we extract of roots presently (110±0.13 mg/100
found that they are absent in roots. Glycoside gm), and a lower content in shoots, flowers and
compounds were present in C. ternatea leaf, seeds. Optimum levels of Phenols (45±0.13
shoot, flower and seed presently which agrees mg/100 gm) and Saponins (2.0±0.6 mg/100
with Salhan et al. (2011). They were however gm) are also reported presently in the roots and
absent in root extracts. these results are similar to those of Chauhan et
Presently, Steroids were present in leaf which al. (2012) and Kaisoon et al. (2011).
confirms the report of Kamilla et al. (2009) and
absent in shoot, flower and seed extracts which CONCLUSION
also confirms the report of Kamilla et al. It is concluded that Clitoria ternatea is a plant
(2009). Saponins were reported in the root of C. with a variety of ethnic medicinal uses. The
ternatea presently which agrees with qualitative analysis of Clitoria ternatea shows
Manalisha and Chandra (2011). Saponins were the presence of bioactive compounds such as
presently absent in shoot and seed extracts Alkaloids, Tannins, Glycosides, Resins,
which agrees with Kamilla et al (2009). Steroids, Saponins, Flavonoids and Phenols.
Traditionally Saponins are extensively used as The quantitative estimation of total Saponins,
detergents, pesticides and molluscicides, in Flavonoids and Phenols in roots and of
addition to their industrial applications as Flavonoids in shoots, flowers and seeds is also
foaming and surface active agents, they also reported which is very important for the
have beneficial health effects (Shi et al. 2004). pharmaceutical industry. This is valuable
Similar to the present findings, Flavonoids information for preparation of drugs in
 P. MANJULA, CH. MOHAN, D. SREEKANTH, B. KEERTHI AND B. PRATHIBHA DEVI 178

pharmaceutical industry and stress the need for 960.

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Morris J B 1999 Legume genetic resources with novel
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