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Antibacterial and Phytochemical Evaluation of Pergularia Daemia from Nagapattinam Region

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Antibacterial and Phytochemical Evaluation of Pergularia Daemia from Nagapattinam Region International Journal of Scientific and Research Publications, Volume 4, Issue 11, November 2014 1 ISSN 2250-3153 Antibacterial And Phytochemical Evaluation of Pergularia daemia from Nagapattinam Region. Vijaya Packirisamy * Vijayalakshmi Krishna Moorthy ** *Department of Biochemistry, Bharathiar University,Coimbatore, Tamilnadu, India. ** Department of Biochemistry,Bharathi women’s College,University of Madras, Tamilnadu, India. Abstract- The present study deals with the phytochemical to treat infantile diarrhoea and malarial intermittent examination and therapeutic importance of Pergularia daemia fevers(Kirtikar. K.R, Basu.B.D,1999; Nadkarani. A.K,1976 ) .The plant Pergularia daemia has been traditionally used as Anonymous,1995). Latex of this plant is used for toothache ( laxative ,antipyretic ,expectorant and also used to treat infantile Hebbar et al 2004). Stem barks are used for cold&fever (Dokosi diarrhea and malarial fever.This study involves the preliminary .O.B, 1998; Bruce .T.B.F, 1998&2000). Aerial parts of this plant phytochemical screening followed by antibactertial activity. The have been reported for the various pharmacological activities phytochemicals such as Tannins ,Saponins ,Flavonoids like hepatoprotectivity (Suresh kumar et al.,2006), ,Quinones, Phenols and Alkaloids,Steroids,Glycosides were antifertility(Golam Sadik et al.,2001), antidiabetic (Wahi et al present in the hydroalcohol of Pergularia daemia leaf extracts 2002) analgesic, antipyretic and anti inflammatory . The was higher amount than the other solvent extracts.In the phytochemical investigation shows the presences of Antimicrobial activity both gram cardenolides, alkaloids and saponins(Sathish et al.,1998). The plant was found to contain various triterpenes and steroidal Index Terms- Phytochemical, Antipyretic, Antibactertial compounds(Anjaneyulu et al.,1998). activity, Pergularia daemia ,MIC. Many antibacterial agents are available in the Market, which are developed by the scientists. But still the microbes are challenging the scientist by developing the resistance to the I. INTRODUCTION presently available drugs. Plants are known to produce a variety he medicinal plants are useful for healing as well as for of compounds to protect themselves against a Variety of Tcuring human diseases because of the presence of the pathogens and therefore Considered as potential source for phytochemical constitutents(Nostro et al.,2000).They are different classes of antimicrobial Substances(Micheal.J, Pelezec, grouped as alkaloids, glycosides ,steroids, coumarins, flavonoids Chan. E.C.S, 1998). So, the present investigation has been and essential oils.Over 50% of all modern clinical drugs are of carried out to shed light on the preliminary phytochemical natural origin(Cordell. G, 1995). Research in medicinal plants Screening and to study the antibacterial activity of Pergularia has gained a renewed focus recently. The prime reason is that daemia leaf extracts. other system of medicine although effective and it has a number of side effects that often lead to serious complications. Plant based system of medicine being natural does not pose serious II. MATERIALS AND METHODS problems. Pergularia daemia also called as Pergularia extensa A. Collection of Plant Sample: The fully mature or Daemia extensa, belongs to milky weed family Pergularia daemia leaves were collected in August – September Asclepiadaceae. Generally, the family Asclepiadaceae includes 2012 from south poigainallur village in Nagapattinam district of more than 2000 species classified under 280 genera are Tamilnadu, India and the Plant was taxonomically identified by distributed world wide in the tropical and subtropical Dr.P.Jayaraman, Plant anatomy research centre, Chennai, regions(Pankaj et al.,2003). Tamilnadu, India. The Voucher Specimen of Pergularia daemia ASCLEPIADACEAE is a large genus of shrubs, and was (PARC/2013/2118). Pergularia daemia (forsk) chiov belongs to this family. B. Processing of plant sample:The leaves of Pergularia Asclepiadaceae is foetid smelling Laticiferous twiner found in daemia are properly washed in tap water and then rinsed in the plains though out the hot pasts of India, ascending to an distilled water. The rinsed leaves are shade dried and powdered altitude of 1000 meters in Himalayas (Anonymous,1997). using electrical blender. The powder was stored in air tight glass Pergularia daemia asclepiadacae (Khare. C.P, 2007) known as containers, protected from Sunlight until required for analysis . Pergularia in English, Veliparuthi in Tamil, Uttaravaruni in Solvent extraction: Sanskrit and Utranajutuka in Hindi belongs to the Pergularia About 10 gm of powdered plant material was soaked Species is a perennial twining herb and is widely distributed in separately in 100ml of methanol, ethanol, hydro alcohol (1:1) , the tropical and subtropical regions of Asia and South Africa ethylacetate, chloroform for 3-4 days at room temperature in dark and have multiple applications in different folk medicines. condition. The extracts were filtered by using what man filter Traditionally the plant Pergularia daemia is used as paper. The filtrate was concentrated to dryness under reduced antihelmintic, Laxative , antipyretic , expectorant and also used pressure at 40 0 C using a rotary evaporator and stored at 4o C for www.ijsrp.org International Journal of Scientific and Research Publications, Volume 4, Issue 11, November 2014 2 ISSN 2250-3153 further use. Each extracts was resuspended in the respective brown ring at the interface indicated the deoxy sugar solvent and used for the qualitative and quantitative analysis of characteristics of cardenolides. A Violet ring may appear below phytochemicals. For the antimicrobial testing the extracts were the ring while in the acetic acid layer, a green ring may be reconstituted to a specific concentration in dimethyl sulphoxide formed (Sofawora. A,1993; SomoLenski. S.J,1974; Harbone. (DMSO). J.B,1973). Aqueous extraction: Glycosides: Glycosides are compounds which upon 10 gm of dried powdered plant leaves was soaked in 100ml hydrolysis give rise to one or more sugars(glycones) and a of distilled water for 3-4 days at room temperature in dark compound which is not a sugar (aglycone or genine).To 1 ml of condition. Then the water extract was filtered through filtered the extract in glacial acetic acid, few drops of ferric chloride and through filter paper and the filtrate was used for the concentrated sulphuric acid was added and observed for a phytochemical analysis. reddish brown colour at the junction of 2 layers and the bluish C. Phytochemical Analysis: green colour in the upper layer) (Siddiqui, A.A et al.1997). The tests were done to find the presence of the active Phenols & Tannins :Crude extract was mixed with 2ml of chemical Constituents such as alkaloids, glycosides, terpenoids, 2% Fecl3 solution. A blue green or black colour indicated the steroids, flavonoids, tannins by the following Procedures. presence of phenols & tannins(26 Sofawora, 1993; Trease. Alkaloids: Alkaloids are basic nitrogenous compounds with G.E,1989; Harborne. J.B,1974). definite physiological and pharmacological activity. Alkaloid Quinones :Dilute sodium hydroxide was added to the 1ml solution produce white yellowish precipitate when a few drops of of crude extract and the blue green or red coloration indicates Mayer’s reagents are added (Siddiqui et al.,1997). Most alkaloids the presence of quinones.( Sofawora, 1993; A, Harbone. are precipitated from neutral or slightly acidic solution by J.B,1973; Trease G.E,1989). Mayer’s reagent(Evans, W.C, Trease,2002). The alcoholic Betacyanin :About 2ml of extract was added with 1ml of extract was evaporated to dryness and the residue was heated on 2N NaOH and heated for 5min. The Formation of bluish green a boiling water bath with 2% hydrochloric acid. After cooling, colour indicates the presence of antho cyanin and Formation of the mixture was filtered and treated with a few drops of mayer’s yellow colour indicates the presence of beta cyanins(Yadav et reagents. The sample were then observed for the presence of al.2011). turbidity or yellow precipitation. D. Bacterial Cultures: Steroids: 1ml of the plant extract was dissolved in 10 ml Gram Positive bacterial cultures like Staphylococcus of chloroform and equal volume of concentrated sulphuric acid aureus, Bacillussubtilis, Staphylococuspyogenes and gram was added by sides of the test tube. The upper layer turns red and negative Organisms like Klebsiella pneumoniae, Aeromonas sulphuric acid layer showed yellow with green fluorescence. This hydrophila, Escherichia coli, Pseudomonas aeruginosa, Vibrio indicated the presence of steroids( Gibbs ,1974). Harveyi are used as test organisms. All the test strains were Terpenoids: To 2ml of the plant extract was added to 2ml maintained on nutrient agar and were sub cultured once in every of acetic anhydride and concentrated H2SO4 . The formation of two weeks. blue green ring indicate the presence of terpenoids(Ayoola et al., Antibacterial assay: 2008). Antibacterial activity of the plant extract was determined Tannins: 2ml of extract was added to few drops of 1% lead using a Kirby- Bauer disk diffusion method( Bauer et al.,1966). acetate , and the yellowish precipitate indicated the presence of Briefly, 100 µl of the test bacteria were grown in 10ml of fresh tannins(Treare, G.E, and Evans W.C ,1985). media until they reached a count of approximately 108 cells of Saponins: 5ml of Extract was mixed with 20ml of distilled bacteria
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