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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.



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


7 1105.21 C-F Stretching Halogen



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



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



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



8 1325.10 C-H Stretching Alkenes

9 1381.03 C-OStretching Primary alchol





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



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



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.

REFERENCES:

1. Venkatesh S, Reddy GO, Reddy BM, Ramesh M, and Appa Rao

AVN. Antihyperglycemic activity of Caralluma attenuates.

Fitoterapia., 2003; 74: 274-279.

2. Chopra RN, Chopra IC. Indigenous drug of India. VN. Dhar

and sons pvt. 501.

3. Caius JE. The medicinal and poisonous plants of India.

Jodhpur, Scientific publishers. 1986; 166-171.

4. Nadkarni AC. Indian materia medica. Edn popular

Prakashan, 1982; 1(3): 362.

5. Abiramasundari P, Priya V, Jeyanthi GP, and Gayathridevi S.

Evaluation of antibacterial activity of Cocculus hirsutus.

Hygeia. J. D. Med., 2011; 3(2): 26-31.

6. Bhavna H, Mariya S, Bothara B. Ethnopharmacological

properties of Cocculus hirsutus Diels- A Review.

International Journal of Pharmaceutical Sciences Review and

Research, 1986; 7(1): 108-112.

7. Praveen K, Dixit, Suchita Mittal. On over view of herbal

antimicrobial agents: Boon for treatment of microbial

infections. IJPRD, 2013; 5(6): 38-45.

8. Sourabh Jain, Manish Lavhale, and Nayak S. Preliminary

phytochemical studies on the roots of Cocculus hirsutus,

LINN. Ancient Science of Life, 2004; 23(3): 42-45.

9. Rakkimuthu R, KavithaKrishna R, Suganyadevi P, Aravinthan

KM. Quantitative phytochemical analysis and their

antioxidant activity of Cocculus hirsutus (1)

Diels.International Journal of Phytomedicine, 2012; 4: 447-

455.

10. Kalirajan A, Savarimuthu Michael J, Ranjit Singh AJA, and

Padmalatha C.Antimicrobial and wound healing studies on

the extracts of the medicinal plant Cocculus hirsutus (LINN).

International Journal of Applied Biology and Pharmaceutical

Technology, 2012; 3(2): 63-67.

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

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