International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

___________________________________________________________________________Research Paper

Phytochemical Anlalysis of the Indian Medicinal Plant Argyreia involucrata

Gutal Valyfathulla Shaik*
* Department of Biotechnology, Acharya nagarjuna university, Guntur, AP, India. ABSTRACT Argyreia involucrata plant was studied for the presence of phytoconstituents. Petroleum ether, acetone, methanol, ethanol and water extracts of leaf and stem materials shows the presence of different phytoconstituents, carbohydrates, cardiac glycosides, phenols, flavonoids, phytosterols, saponins and fixed oils. However the leaf and stem solvent extracts do not shows the presence of alkaloids. Keywords: Argyreia involucrata, soxhlet apparatus, phytoconstituents INTRODUCTION Medicinal plants are considerably useful and economically essential. They contain the active constituents that are used in the treatment of many human diseases1. The demand for more and more drugs from plants is continuously increasing. It is therefore essential to evaluate plants of medicinal value systematically for various ailments that are used in traditional medicine. Plants used in traditional medicine contain a vast array of substances that can be used to treat chronic and infectious diseases2. Secondary metabolites serve as plant defense mechanism against predation by microorganisms, insects and herbivores. Some, such as terpenoids, give plants their odors; others (quinines and tannins) are responsible for plant flavor and some of the herbs and spices used by humans to season food yield useful medicinal compounds3. In recent years, secondary plant metabolites (photochemical), previously with unknown activities, have been extensively investigated as a source of medicinal agents4. A wide range of medicinal plant parts is used to extract as raw drugs and they possess varied medicinal properties5. The Convolvulaceae family commonly known as morning glory family consists of 85 genera and 2,800 species. They are mostly twining herbs, shrubs, sometimes with milky sap. They are categorized by the solitary (or) terminal (or) axillary flowers. Some species of this family consists of ergotine alkaloids and cyanogenic glycosides commonly used as ingredients in psychoactive drugs. Genus Argyreia consists of approximately 100 species of rank growing woody climbers of tropical Asia to Australia. They consist of silvery leaves _______________________________________ *Address for correspondence: E-mail: [email protected] and showy purple flowers. It belongs to the tribe Argyreiae. They are closely allied to the morning glory genus Ipomoea. Argyreia nervosa commonly called as Babywood rose is popular for its decorative dried fruit, traded as wood rose. The present plant Argyreia involucrata was commonly used in folk medicine. The purpose of the present study was to know the different secondary metabolites (phytoconstituents) present. MATERIALS AND METHODS Plant material The plant Argyreia involucrata was collected from the Madhukarai hills, Coimbatore District, Tamil Nadu. The plant was taxonomically identified by Dr. S. Karuppaswamy, Department of Botany, Botanical Research Centre, Madura College, Madurai 11, Tamil Nadu, South India. The fresh plant parts were collected. The leaves and stems were detached and washed with clean water. Plant materials were air dried on a clean sheet for one week at room temperature. Preparation of plant extract The dried leaves and stems of Argyreia involucrata was powdered and passed through sieve #10. The fine powder was stored in airtight containers and stored. Extraction was carried out using a Soxhlet apparatus at room temperature under normal conditions. 10 gms of the sieved powder was weighed accurately and subject to extraction at room temperature using petroleum ether, acetone, methanol, ethanol and water successively. Before extraction with the next solvent the powder was air dried to remove the adhering solvent. The extract obtained was filtered, concentrated and stored in air tight containers for further studies.

Vol. 2(4) Oct - Dec 2011

www.ijrpbsonline.com

1778

International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

PHYTOCHEMICAL EVALUATION Phytochemical examinations has to be carried out for all the extracts as per the standard methods6 Detection of alkaloids: Extracts have to dissolve individually in dilute Hydrochloric acid and filtered. The filtrates have to be used to test for the presence of alkaloids. a) Mayers Test: Filtrates has to be treated with Mayers reagent (Potassium Mercuric iodide). Formation of a yellow cream precipitate indicates the presence of Alkaloids. b) Wagners Test: Filtrates has to be treated with Wagners reagent (Iodine in potassium iodide). The formation of a brown / reddish brown precipitate indicates the presence of Alkaloids. c) Dragendroffs test: Filtrates has to be treated with Dragendroffs reagent (solution of potassium bismuth iodide). Formation of a red precipitate indicates the presence of Alkaloids. d) Hagers test: Filtrates has to be treated with Hagers reagent (saturated picric acid solution). Formation of a yellow colored precipitate indicated the presence of Alkaloids. Detection of carbohydrates: Extracts has to be dissolved individually in 5 ml distilled water and filtered. The filtrates were used to test for the presence of carbohydrates. a) Molischs test: Filtrates has to be treated with 2 drops of alcoholic -naphthol solution in a test tube and 2 ml. of conc. Sulphuric acid was added carefully along the sides of the test tube. Formation of violet ring at the junction indicates the presence of carbohydrates. b) Benedicts test: Filtrates has to be treated with Benedicts reagent and heated in a water bath. Formation of an orange red precipitate indicates the presence of reducing sugars. c) Fehlings test: Filtrates has to be hydrolysed with dil. HCl, neutralized with alkali and heated with Fehlings A & B solutions. Formation of a red precipitate indicates the presence of reducing sugars. Detection of glycosides: Extracts have to be hydrolysed with dil. HCl, and then subjected to test for glycosides. a) Modified Borntragers Test: Extracts have to be treated with a Ferric Chloride solution and immersed in boiling water for about 5 minutes. The mixture was cooled and shaken with an equal volume of benzene. The benzene layer was separated and treated with ammonia solution. Formation of rose-pink

colour in the ammonical layer indicates the presence of anthranol glycosides. b) Legals Test: Extracts have to be treated with sodium nitroprusside in pyridine and methanolic alkali. Formation of pink to blood red colour indicates the presence of cardiac glycosides. c) Keller-Killiani test: Extracts have to be treated with 1 ml glacial acetic acid + FeCl3 + concentrated sulphuric acid. Formation of green blue colour indicates the presence of cardiac glycosides. Detection of saponins: a) Froth Test: Extracts have to be diluted with distilled water to 20 ml and this was shaken in a graduated cylinder for 15 minutes. Formation of 1 cm layer of foam indicates the presence of saponins. b) Foam Test: Small amount of extract has to be shaken with little quantity of water. If foam produced persists for ten minutes it indicates the presence of saponins. Detection of phytosterols: a) Salkowskis Test: Extracts have to be treated with chloroform and filtered. The filtrates were treated with a few drops of conc. Sulphuric acid, shaken and allowed to stand. The appearance of golden yellow color indicates the presence of triterpenes. b) Liebermann Burchards Test: Extracts have to be treated with chloroform and filtered. The filtrates were treated with a few drops of acetic anhydride, boiled and cooled. Concentrated sulphuric acid was added carefully along the sides of the test tube. The formation of brown ring at the junction Indicates the presence of phytosterols. c) Tshugajeu test: Extracts have to be treated with chloroform and filtered. Excess of acetyl chloride and a pinch of Zinc Chloride was added, kept aside for sometime till the reaction was complete and then warmed on water bath. Appearance of eosin red colour indicates the presence of triterpenes. Detection of fixed oils & fats: a) Stain Test: Small quantities of extracts have to be pressed between two filter papers. An oily stain on filter paper indicates the presence of fixed oil. Detection of resins: a) Acetone-water Test: Extracts has to be treated with acetone. Small amount of water was added and shaken. The appearance of turbidity indicates the presence of resins. Detection of phenols: a) Ferric Chloride Test: Extracts have to be treated with a few drops of ferric chloride

Vol. 2(4) Oct - Dec 2011

www.ijrpbsonline.com

1779

International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

solution. Formation of bluish black color indicates the presence of phenols. Detection of tannins: a) Gelatin Test: To the extract, 1% gelatin solution containing sodium chloride has to be added. Formation of white precipitate indicates the presence of tannins. Detection of flavonoids: a) Alkaline Reagent Test: Extracts have to be treated with a few drops of sodium hydroxide solution. Formation of intense yellow color, which becomes colorless on the addition of dilute acid, indicates the presence of falvonoids. b) Lead acetate Test: Extracts have to be treated with a few drops of lead acetate solution. Formation of a yellow color precipitate indicates the presence of flavonoids. c) Shinoda Test: To the alcoholic solution of extracts, a few fragments of magnesium ribbon and concentrated HCl has to be added. The appearance of magenta color after a few minutes indicates the presence of flavonoids. Detection of proteins and amino acids: a) Xanthoproteic Test: The extracts have to be treated with a few drops of concentrated Nitric acid solution. Formation of yellow color indicates the presence of proteins. b) Ninhydrin Test: To the extract, 0.25% ninhydrin reagent has to be added and boiled for a few minutes. Formation of blue color indicates the presence of amino acids. c) Biuret Test: The extracts have to be treated with 1 ml of 10% sodium hydroxide solution and heated. To this a drop of 0.7% copper sulphate solution was added. Formation of

purplish violet color indicates the presence of proteins.

Detection of diterpenes: a) Copper acetate Test: Extracts have to be dissolved in water and treated with a few drops of copper acetate solution. Formation emerald green color indicates the presence of diterpenes. b) RESULTS AND DISCUSSION Preliminary phytochemical analysis of the leaf and stem extracts indicates the presence of carbohydrates, cardiac glycosides, phytosterols, phenols, falvonoids, diterpenes and saponins (Table 1,2). The type of phytoconstituents of the leaf and stem solvent and water extracts were not similar indicating the variation. CONCLUSION The leaf and stem organic solvent and water extracts showed the presence of phytochemical constituents. Further studies have to be carried out to reveal the activities of these phytoconstituents present in this plant material ( Table 1,2). ACKNOWLEDGEMENT I am thankful to all my Gutal family members for their support of the present work. I am thankful to Dr. J.V.Prabhakara Rao, vice-chancellor, Naik, Registrar of Rayalaseema University, Kurnool for encouraging and providing the lab facilities in Department of Biotechnology.

Vol. 2(4) Oct - Dec 2011

www.ijrpbsonline.com

1780

International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

Table 1. Preliminary phytochemical analysis of leaf material of the plant Argyreia involucrata
Phytoconstituent Test Mayers Test Wagners Test Dragendroffs test Hagers test Molischs test Benedicts test Fehlings test Modified Borntragers Test Legals Test Keller-Killiani test Froth Test Foam Test Salkowskis Test Liebermann Burchards Test Tshugajeu test Stain Test Acetone-water Test Ferric Chloride Test Gelatin Test Alkaline Reagent Test FLAVONOIDS Lead acetate Test Shinoda Test Xanthoproteic Test Ninhydrin Test Biuret Test Copper acetate Test Ether extract + + + + + Acetonic extract + + + + + Methanolic extract + + + + + + Ethanolic extract + + + + + + + + Aqueous extract + + + + + + +

ALKALOIDS

CARBOHYDRATES

GLYCOSIDES

SAPONINS

PHYTOSTEROLS FATS & OILS RESINS PHENOLS TANNINS

PROTEINS & AMINOACIDS DITERPENES

+ = present - = absent

Vol. 2(4) Oct - Dec 2011

www.ijrpbsonline.com

1781

International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

Table 2. Preliminary phytochemical analysis of stem material of the plant Argyreia involucrata
Phytoconstituent Test Mayers Test Wagners Test Dragendroffs test Hagers test Molischs test Benedicts test Fehlings test Modified Borntragers Test Legals Test Keller-Killiani test Froth Test Foam Test Salkowskis Test Liebermann Burchards Test Tshugajeu test Stain Test Acetone-water Test Ferric Chloride Test Gelatin Test Alkaline Reagent Test FLAVONOIDS Lead acetate Test Shinoda Test Xanthoproteic Test PROTEINS & AMINOACIDS DITERPENES Ninhydrin Test Biuret Test Copper acetate Test + + + + + + + + + + + + + + Ether extract + + + + Acetonic extract + + + + Methanolic extract + + + + + + + + Ethanolic extract + Aqueous extract + + + + +

ALKALOIDS

CARBOHYDRATES

GLYCOSIDES

SAPONINS

PHYTOSTEROLS FATS & OILS RESINS PHENOLS TANNINS

+ = present - = absent REFERENCES 1. Stary, F., Hans, S. (1998). The national guides to medical herbs and plants. Tiger Books. Int. Plc. UK. Sumathi, P., Parvathi, A. (2010). Antimicrobial activity of traditional medicinal plants. J. medicinal plants research, Vol 4(4): 316 321 . Cowan, M.M. (1999). Plant products as Antimicrobial agents. Clinical Microbiology Reviews : 564 582. Krishnaraju, A.V., Rao, T.V.N. and Sundararaju D et al., (2005). Assessment of bioactivity of Indian medicinal plants using Brine shrimp (Artemia salina) lethality assay. Int. J. Appl. Sci. Eng 2 : 125 134. Uniyal, S.K., Singh, K.N., Jamwal. P. and Lal, B. (2006). Traditional use of medicinal plants among the tribal communities of Unhota Bhangal, Western Himalayan. J. Ethnobiol. Ethnomed., 2 : 1 14. Brain, K.R., Turner, T.D. (1975). The practical evaluation of phytopharmaceuticals. 2nd ed. Bristol : Wright sciencetechnica : 81 82.

5.

2.

3.

6.

4.

Vol. 2(4) Oct - Dec 2011

www.ijrpbsonline.com

1782