Content uploaded by Aparna Karthikeyan
Author content
All content in this area was uploaded by Aparna Karthikeyan on Mar 12, 2018
Content may be subject to copyright.
Int. J. Pharm. Sci. Rev. Res., 25(1), Mar – Apr 2014; Article No. 52, Pages: 307-309 ISSN 0976 – 044X
International Journal of Pharmaceutical Sciences Review and Research
Available online at
www.globalresearchonline.net
307
Joyce Priyakumari C*, Aparna K, Sagaya Jansi R, Gladies Kezia
Bioinformatics Centre of BTISnet, Madras Christian College, Chennai, Tamil Nadu, 600059, India.
*Corresponding author’s E-mail: joycebenezer@gmail.com
Accepted on: 05-02-2014; Finalized on: 28-02-2014.
ABSTRACT
Phytochemical analysis of extract of Didymocarpus pedicellatus was conducted to detect the presence of various secondary
metabolites. These plants are widely used in South Indian Cuisine. It is commonly known as Black stone flower. Didymocarpus
pedicellatus has a strong ethanopharamaceutical background. D. pedicellatus revealed the presence of Alkaloids, flavonoids, tannis,
phenols etc.
Keywords: Didymocarpus pedicellatus, Phytochemical extraction, Secondary metabolites.
INTRODUCTION
ndian spices have many medicinal properties.
Didymocarpus pedicellatus commonly called as Kalpasi
in Tamil is widely used in Indian Cuisine. Didymocarpus
pedicellatus (Kalpasi) is a valuable although a lesser
known medicinal plant. It is popularly known as black
stone flower and is a rare flower in the family
Gesneriaceae. The plant is native to Tropical Asia.1In
Ayurveda it is known as shilapushpa, shantapushpi and
sometimes pasanbheda.2 It is often dried and used as an
Indian spice. Black stone flowers have many medicinal
properties. They are a good pain reliever and also
promote early healing of wounds. It helps in digestion and
suppresses the respiratory disorders. In this study we are
trying to find the phytochemical properties from the dried
flowers of Didymocarpus pedicellatus.
Traditionally D. pedicellatusis used in the treatment of
renal diseases particularly kidney stones.3 It also regulates
calcium absorption in the body. The plant is known for its
diuretic effect and in maintaining healthy urinary tract.4
Phytochemical studies have attracted the attention of
plant scientists due to the development of new
sophisticated techniques. These techniques played a
significant role in giving the solution to systematic
problems on the one hand and in the search for
additional resources of raw materials for pharmaceutical
industry on the other hand.5
Medicinal plants contain some organic compounds which
provide definite physiological action on the human body
and these bioactive substances include tannins, alkaloids,
carbohydrates, Terpenoids, steroids and flavonoids.6,7
D. pedicellatus extract was found to possess a high
content of total polyphenolics, exhibit potent reducing
power and significantly scavenge free radicals including
several reactive oxygen species (ROS) and reactive
nitrogen species (RNS). The extract also significantly and
dose-dependently protected against Fe-NTA plus H2O2-
mediated damage to lipids and DNA. 8
MATERIALS AND METHODS
Preparation of plant extract
Dried flowers of Didymocarpus pedicellatus were
collected and homogenized into a fine powder using
mechanical grinder.
Preparation of Extracts
Powdered plant material was subjected to successive
solvent extract using solvents like Ethanol, Methanol,
Chloroform, and Acetone. 20g of powdered flower
sample was added to 200 ml of suitable solvents and the
crude was extracted after 48 hrs. The extracts were
filtered and evaporated. Phytochemical analyses were
carried out from the crude.
Phytochemical analysis
Phytochemical analysis were done by certain reagents to
test the alkaloid, carbohydrates, reducing sugar,
flavonoids, glycosides, phenolic compounds, saponins,
steroids, amino acids and tannins.
Test for Alkaloids
Extracts were mixed with dilute hydrochloric acid and
following tests were done.
a. Mayer’s Test
Extracts were treated with Mayer’s Reagent (Mercuric
chloride & Potassium iodide). Formation of a cream
indicates the presence of alkaloids.
b. Wagner’s Test
Extracts were treated with Wagner’s Reagent (Iodine &
Potassium iodide). Formation of a reddish brown
precipitate indicates the presence of alkaloids.
Test for Flavonoids
Aqueous extracts was treated with dilute ammonia and
conc. Sulphuric acid was added. Yellow colouration
indicates the presence of flavonoids.
Phytochemical Analysis of Didymocarpus pedicellatus
I
Research
Article
Int. J. Pharm. Sci. Rev. Res., 25(1), Mar – Apr 2014; Article No. 52, Pages: 307-309 ISSN 0976 – 044X
International Journal of Pharmaceutical Sciences Review and Research
Available online at
www.globalresearchonline.net
308
Test for Terpenoids
To the extracts 2ml of chloroform was added, conc.
Sulphuric acid was carefully added to form a layer.
Reddish brown colouration at the interface indicates the
presence of terpenoids.
Test for Steroids
a. Extracts were mixed with 2ml of chloroform and
conc.sulphuric acid was added sideways. A red colour
producing at the lower chloroform layer indicate the
presence of steroids.
b. Extracts were mixed with 2ml of chloroform, 2 ml of
conc.sulphuric acid and 2ml of acetic acid were poured
into the mixture. Development of greenish colouration
indicates the presence of steroid.
Test for Phenols
Ferric chloride Test
Extracts were treated with 3 to 4 drops of ferric chloride
solution. Formation of bluish black colour indicates the
presence of phenol.
Test for Glycosides
a. Libermann Burchard’s Test
Extracts were treated with 2 ml of chloroform and 2 ml of
acetic acid. The mixture was cooled in ice and conc.
Sulphuric acid was added. Colour change from violet to
blue to green the indicates the presence of glycosides
b. Salkowski’s Test
Extracts were mixed with 2ml of chloroform. Then 2 ml of
conc. Sulphuric acid was added carefully and shaken
gently. A reddish brown colour indicates the presence of
steroidal ring.
c. Keller-kilani Test
Extracts were mixed with 2ml of glacial acetic acid
containing 1 to 2 drops of ferric chloride solution. The
mixture was then poured into another test tube
containing 2 ml of conc. Sulphuric acid. A brown ring at
the interface indicates the presence of cardiac glycosides.
Test for Protein
a. Ninhydrin
Extracts wercxdfdfe boiled with 2 ml of ninhydrin
solution. Appearance of violet colour indicates the
presence of proteins.
b. Xanthoproteic
Extracts were treated with few drops of conc. Nitric acid.
Formation of yellow colour indicates the presence of
proteins.
Teat for Carbohydrates
a. Benedict’s solution
Extracts were treated with 2 ml of Benedict’s reagent and
gently heated. Formation of orange red precipitate
indicates the presence of reducing sugar.9
b. Fehling’s Test
Equal volume of Fehling A and Fehling B reagents were
mixed together and 2 ml of it was added to the extract
and gently boiled. A brick red colour indicates the
presence of reducing sugar.
c. Iodine Test
Extracts were mixed with 2 ml of Iodine solution, a dark
blue or purple colour indicates the presence of
Carbohydrates.
Test for Tannins
Extracts were boiled with 10 ml of water in a test tubes. A
few drops of ferric chloride was added and observed for
blue black colouration.
Test for Saponins
Extracts were boiled with distilled water. The solution was
shaken vigorously for a stable, persistent, froth or foam.
Table 1: Phytochemical Screening of Didymocarpus
pedicellatus
Tests Ethanolic
Extract Acetone
Extract Methanol
Extract Chloroform
Extract
Alkaloids
Mayer’s test + + + +
Wagner’s Test + + + +
Glycosides
Liberman’s Test + + - -
Salkowski’s Test + + + +
Keller Kilani’s Test + + + +
Carbohydrates
Fehling’s Test + + + +
Benedict’s Test + + + +
Iodine Test + + + +
Proteins
Xanthoproteic
Test + + + +
Ninhydrin Test + + + +
Tannins + - + +
Flavonoids + + + +
Steroids + - + +
Terpenoids + + + +
Phenol + + + +
Saponin + + + +
Int. J. Pharm. Sci. Rev. Res., 25(1), Mar – Apr 2014; Article No. 52, Pages: 307-309 ISSN 0976 – 044X
International Journal of Pharmaceutical Sciences Review and Research
Available online at
www.globalresearchonline.net
309
RESULTS AND DISCUSSION
The phytochemical active compounds of Didymocarpus
pedicellatus were qualitatively analyzed for flowers and
the results are presented.
The phytochemical studies of four solvents shows the
presence of flavonoids, steroids, terpenoids, saponins,
tannins, proteins, phenols, alkaloids, glycosides and
carbohydrates in almost all the four extracts. Different
phytochemical have been found to possess a wide range
of activities, which may help in protection against chronic
diseases. D. pedicellatus contains Chalcones10,
Polyterpenes like didymocarpol, didymacarpenol.4 Since
D. pedicellatus contains flavonoids, it may be also valued
for anticancer activity.
Results obtained from phytochemical screening were
shown in Table.1. Thus the present investigation revealed
that D. pedicellatus appears to be rich in secondary
metabolites, the preliminary study shows the presence of
bioactive compounds. Further studies are in progress to
isolate the active compounds.
Acknowledgements: This work is supported by BTIS net
centre sponsored by Department of Biotechnology,
Ministry of Science & Technology, and Government of
India.
REFERENCES
1. McGuffin M, Herbs of commerce, Second Edition 2000.
2. Bahl CP and Seshadri TR, Pashanbhedi: drugs for urinary
calculus, Eds.1978., K.N. Udupa, 7 7–98.
3. Kapoor SL, Kapoor LD,On the botany and distribution of
`pashanbheda'. Sachitra Ayurved, 28(12), 1976, 769–791.
4. Singh AP, A Short review on Didymocarpus pedicellata:
The Lithontriptic Ethnomedicine, Ethnobotanical Leaflets,
11, 2007, 73–75.
5. Mojab F, Kamalinejad M, Naysanch Ghaderi, Hamid
Rozavahidipour, Phytochemical screening of some species
of Iranian plants. Iranian Journal of Pharmaceutical
Research, 2:2, 2003, 77–82.
6. EdogaHO, Okwu DE, Mbaebie BO, Phytochemical
constituents of some Nigerian medicinal plants, African
Journal of Biotechnology, 4:7, 2005, 685–688.
7. Mann J, Secondary Metabolism. Oxford University press,
London, 1978.
8. Kaur G, Lone IA, Athar M, Protective effect of
Didymocarpus pdeicellata on ferric nitrilotriacetate (Fe-
NTA) induced renal oxidative stress and hyperproliferative
response, Chemico Biological Interaction, 165:1, 2007,
33–44.
9. Benedict SR: A Reagent for the detection of reducing
sugars, The Journal of biological Chemistry, 5, 1909, 485–
487.
10. Rathore JS, Garg SK, Gupta SR, A chalcone and flavones
from Didymocarpus pedicellata. Phytochemistry, 20, 1981,
1755-1756.
Source of Support: Nil, Conflict of Interest: None.