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Packialkshmi N et al., J. Sci. Res. Phar. 2014, 3(2), 72-75
Journal of Scientific Research in Pharmacy 2014, 3(2) 72-75
Journal of Scientific Research in Pharmacy Research Article Available online through ISSN: 2277-9469
www.jsrponline.com
Preliminary Phytochemical studies on Leaves and Stem of Cocculus Hirsutus, Linn.
Packialakshmi N and Fazila Beevi H PG and Research Department of Microbiology, Jamal Mohamed College (autonomous), Tiruchirappalli – 620 020, Tamil Nadu, India.
Received on: 26-06-2014; Revised and Accepted on: 02-07-2014
ABSTRACT
The leaves of Cocculus hirsutus (Linn) was analyzed for preliminary phytochemical studies with extract of water. The plant is well
reputed in traditional system of medicine ,present studies will help in further validation and standardization of the plant. Phytochemical studies
indicated that the leaf contain a broad spectrum of secondary metabolites,carbohydrates,alkaloids,phenols tannins, fixed oil & fats, gums &
mucilages,flavonoids ,sterols, protein & amino acids.
Key words: Cocculus hirsutus, Leaves, Stem, Minispermaceae, Phytochemicals, Aqueous extract, IR Spectrum.
INTRODUCTION
Herbal drugs are prescribed widely because of their
effectiveness, less side effects and relatively low cost [1]. Therefore
investigation on some active principles from traditional medicinal
plants has become more important. Cocculus hirsutus, known well as
Broom creeper, is found in moderately cool and hot regions of India
particularly Tamil nadu, Bihar and Punjab. Cocculus hirsutus Linn
(Minispermeaceae) is commonly known as Jal-Jammi [2]. In Tamil
this plant is known as Kattukkodi.Indian tribes use various plant
parts of this plant for a wide range of ailments including
constipation, kidney problems [3]. In Tamil nadu kaani tribes of
karaiyar using this plant for the treatment of skin disease, sexual
debility, wound healing, and for the treatment of rheumatism,
tuberculosis, leprosy, dyspepsia, pruritis, flatulence, laxative,
antipyretic and leaves are used in biliousness, eczema, gonorrhea,
opthalmia, and neuralgia. The juice of leaves coagulates in water and
forms mucilage, which is used externally as a cooling and soothing
agent in eczema and impetigo [4]. A number of phytoconstituents
including alkaloids, flavonoids, carbohydrate, oil & fats , phenolic
compounds, protein & amino acids and gums & mucilage from the
plant [5-7].
MATERIALS AND METHOD
Collection of Plant Materials:
The plant Cocculus hirsutus was collected from region of
Tiruchirappalli district and identified by Rapinat Herbarium,
St.Josephs College, Tiruchirappalli. The leaves were separated from
the collected plant and dried under shade. After drying, it was
powdered and used for our studies.
Preliminary Phytochemical Screening:
Phytochemical screening was carried out to assess the
qualitative chemical composition of crude extracts using commonly
employed precipitation and coloration to identify the major natural
chemical groups such as alkaloids, flavonoids, tannins,
saponins,sterols, glycocides, carbohydrate, phenolic compounds, oil
& fats, protein & amino acids, gums & mucilage. General reactions in
this analysis revealed the presence or absence of these compounds
in the crude extracts tested.
*Corresponding author: Dr. Packialakshmi N PG and Research Department of Microbiology,
Jamal Mohamed College (autonomous), Tiruchirappalli–620 020,
Tamil Nadu, India.
*E-Mail: [email protected]
Test for Alkaloids:
Mayer’s Test:
1 ml of plant extract was mixed with small amount of
dilute hydrochloric acid and 1 ml of Mayer’s reagents. Formation of
precipitate indicates the presence of alkaloid.
Test for Flavonoids:
Colour with ferric chloride:
1 ml of extract was taken and a few drops of very dilute
solution of ferric chloride were added. The colour changed to pale
green or red brown colour which indicates the presence of
flavonoids.
Test for Saponins:
Foam Test:
1 ml of extract was diluted separately with distilled water
to 20 ml and shaken with graduated cylinder for 15 minutes.
Formation of lather indicates presence of saponins.
Test for Carbohydrate:
Molisch’s Test:
2 ml of plant extract was dissolved separately in 4 ml of
distilled water and filtered. The filtrate was subjected to Molisch’s
test. Formation of reddish brown ring indicates the presence of
carbohydrate.
Test for Tannins:
Lead acetate Test:
5 ml of extract was added with 1 ml of lead acetate
solution. Flocculent brown precipitate indicates the presence of
tannins.
Test for Sterols:
Liebermann Burchard reaction:
2 ml of plant extract and a few crystal of sodium nitrate
were taken in a dry test tube and heated gently for a minute. It was
cooled and 0.5 ml of concentrated H2SO4 was added. Formation of
orange or pale orange indicates presence of sterols.
Test for Glycosides:
2 ml of plant extract was hydrolyzed with hydrochloric
acid for few hours on a water bath and the hydrolysate was
subjected to legal’s test to detect the presence of different
glycosides.
Legal’s Test:
To the hydrolysate 1 ml of sodium nitroprusside solution
was added and then it was made alkaline with sodium hydroxide
solution. If the extract produced pink to red colour , it indicates the
presence of glycosides.
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Packialkshmi N et al., J. Sci. Res. Phar. 2014, 3(2), 72-75
Journal of Scientific Research in Pharmacy 2014, 3(2) 72-75
Test for Fixed Oils & Fats:
Spot Test:
Few drops of 0.5 N alcoholic potassium hydroxide were
added to small quantity of various extract along with a drop of
phenolphthalein. The mixture was heated on a water bath for 1-2
hrs.Formation of soaps or particle neutralization of alkali indicates
the presence of fixed oil & fats.
Test for Phenolic Compounds:
Ferric chloride Test:
2 ml of plant extracts were taken for the presence of
phenolic compounds with dilute ferric chloride solution (5%) which
gives violet colour.
Test for Gums & Mucilage:
Alcoholic precipitation:
10 ml of extract was added to 25 ml of absolute alcohol
with stirring and filtered. The precipitate was dried in air and
examined for its swelling properties and for the presence of gums &
mucilage.
Test for Protein & Aminoacids:
Biuret Test:
2 ml of extract was dissolved in few ml of water and
treated with ninhydrin reagent gives purple colour.
IR Spectrum Analysis:
FTIR relies on the fact that the most molecules absorb
light in the infra-red region of the electromagnetic spectrum. This
absorption corresponds specifically to the bonds present in the
molecule.The frequency ranges are measured as wave numbers
typically over the range 4000-600 cm-1.
RESULTS
Phytochemical Evalution:
Phytochemical analysis of the aqueous extract indicated
that the leaf and stem contain a broad spectrum of secondary
metabolites, carbohydrates, alkaloids, flavonoids, tannins, sterols,
glycosides, fixed oil & fats, phenolic compounds, gums & mucilage,
protein and amino acid were predominantly found in aqueous
extract of leaf and stem.Saponin was absent in leaves of Cocculus
hirsutus. Saponin and carbohydrates are absent in the stem of
Cocculus hirsutus. (Table. 1).
Infrared Spectrum Analysis:
In present study Table-2, Fig 1 & Table-3, Fig 2 showed
that analysis of Infrared spectrum by Cocculus hirsutus leaves and
stem powder (crude).
Table No. 1: Results of Preliminary Phytochemical screening of aqueous leaf and stem extracts of Cocculus hirsutus
S. No. Name of the Compounds Name of the Test Status of Substance (Aqueous Extract)
Leaves Stem
1 Alkaloids Mayer’s Test + +
2 Flavonoids Ferric chloride Test + +
3 Saponin Foam Test - -
4 Carbohydrate Molisch’s Test + -
5 Tannins Lead acetate Test + +
6 Sterols Libermann Burchard reaction + +
7 Glycosides Legal’s Test + +
8 Fixed oil & Fats Spot Test + +
9 Phenolic compounds Ferric chloride Test + +
10 Gums & mucilage Alcoholic Precipitation + +
11 Protein & Amino acid Biuret Test + +
(+) indicates Presence , (-) indicates Absence
Table No. 2: Infrared spectrum analysis by Cocculus hirsutus leaf powder (crude)
S. No Peak Value Stretching Interpretation
1 470.63 - Benzene
2 534.28 C-Br Stretching Bromine
3 671.23 C-Br Stretching Bromine
4 779.24 C=C Stretching Hydro carbon
5 1022.27 C-O Stretching Ethers
6 1047.35 C-O Stretching Ethers
7 1105.21 C-F Stretching Halogen
8 1149.57 C-O Stretching Ethers
9 1244.09 C-O Stretching Ethers
10 1323.17 N=O Stretching Nitrogroup
11 1381.03 C-O Stretching Phenols
12 1404.18 C-O Stretching Phenols
13 1440.83 C-C Stretching Aromatics
14 1525.69 N=O Stretching Nitrogroup
15 1631.78 C=C Stretching Alkenes
16 1635.00 C=O Stretching Alkenes
17 1730.15 C=O Stretching Esters
18 1807.30 C=O Stretching Anhydrides
19 1853.59 C=O Stretching Anhydrides
20 1876.74 C=O Stretching Esters
21 1901.81 C=O Stretching Carboxylic acid
22 1930.74 C=O Stretching Carboxylic acid
23 2370.51 C=C Stretching Alkenes
24 2852.72 O-H Stretching Carboxylic acid
25 2922.16 O-H Stretching Carboxylic acid
26 3408.22 N-H Stretching Amides
27 3697.54 O-H Stretching Free OH group
28 3782.41 N-H Stretching Amines
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Packialkshmi N et al., J. Sci. Res. Phar. 2014, 3(2), 72-75
Journal of Scientific Research in Pharmacy 2014, 3(2) 72-75
Table No. 3: Infrared spectrum analysis by Cocculus hirsutus stems powder (crude)
S. No Peak Value Stretching Interpretation
1 493.78 - Benzene
2 536.21 C-Br Stretching Halogen
3 673.16 C-Cl Stretching Chlorine
4 1033.85 C-F Stretching Fluorine
5 1107.14 C-F Stretching Fluorine
6 1157.29 C-O Stretching Ethers
7 1246.02 C-O Stretching Ethers
8 1325.10 C-H Stretching Alkenes
9 1381.03 C-OStretching Primary alchol
10 1442.75 C-C Stretching Aromatics
11 1463.97 C-C Stretching Aromatics
12 1527.62 N-H Stretching Amides
13 1546.91 N-H Stretching Amides
14 1600.92 C-C Stretching Alkenes
15 1658.78 N=O Stretching Nitrocompounds
16 1724.36 C=O Stretching Aldehyde
17 1757.15 C=O Stretching Esters&Lactones
18 1811.16 C=O Stretching Anhydrides
19 1876.74 C=O Stretching Esters
20 1930.74 C-O Stretching Carboxylic acid
21 2268.29 C=C Stretching Alkynes
22 2374.37 C=C Stretching Alkynes
23 2854.65 C-H Stretching Alkanes
24 2924.09 C-H Stretching Alkanes
25 3429.43 N-H Stretching Chelatecompounds
26 3697.54 O-H Stretching Free O-H group
27 3784.34 N-H Stretching Amines
Fig. 1: IR Spectrum Analysis of Leaves of Cocculus Hirsutus
Fig. 2: IR Spectrum Analysis of Stem of Cocculus Hirsutus
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Packialkshmi N et al., J. Sci. Res. Phar. 2014, 3(2), 72-75
Journal of Scientific Research in Pharmacy 2014, 3(2) 72-75
DISCUSSION
In the earlier study [8] reported that the roots of Cocculus
hirsutus, LINN, and have been used as an effective drug for analgesic,
anti-inflammatory, anti microbial, hypoglycemic and cardio tonic
with the presence of ash, moisture content, resins, tannins, alkaloid,
lipids, and absence of glycoside, volatile oil, gums and mucilage in
various solvent extracts .The earlier study[9] studied that the fruits of
Cocculus hirsutus have been shown antioxidant activity and the
major source of flavonoids, anthocyanin, phenolic compounds in the
methonolic extract of Cocculus hirsutus. In previous study [10]
reported that the leaves of Cocculus hirsutus shown the presence of
alkaloids, steroids, saponins and tannins.
Knowledge of the phytochemical constituents of plant is
desirable, not only for the discovery of therapeutic agents, but also
because such information may be of value in disclosing new sources
of such economic materials as tannins, oils & fats, gums, flavonoids,
carbohydrate, glycosides precursors for the synthesis of complex
chemical substances. In our study the phytochemical screening of
aqueous extracts of leaf and stem indicate the presence of alkaloids,
flavonoids, carbohydrate, tannins, sterols, glycosides, oil&fats,
phenolic compounds, protein&aminoacid and gums&mucilage.
CONCLUSION
The present study of the plant Cocculus hirsutus showed major
source of phytoconstituents, Hence leaf and stem extract of Cocculus
hirsutus is highly recommended for herbal preparations to the
traditional medicinal practioners and for the pharmaceutical
industries for the mass scale extraction of therapeutic agents.
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How to cite this article:
Packialakshmi N and Fazila Beevi H: Preliminary Phytochemical studies on Leaves and Stem of Cocculus Hirsutus, Linn. J. Sci.
Res. Phar, 2014; 3(2): 72-75.
Conflict of interest: The authors have declared that no conflict of interest exists.
Source of support: Nil