Evaluation Of In-vitro Antibacterial And Anthelmintic .

International Journal of Pharmacognosy and Chinese MedicineISSN: 2576-4772Evaluation of In-vitro Antibacterial and Anthelmintic Activitiesof Sauropus androgynus (Phyllanthaceae) Plant ExtractsPalaksha MN1*, Aishwarya KS2, Arpitha SM2, Pavan Kumar2,Raghu kumar1Department2IVHM2 andKoushikM2Research ArticleVolume 3 Issue 2of pharmacology, Bharathi College of pharmacy, IndiaB. Pharm students, Bharathi College of pharmacy, IndiaReceived Date: May 02, 2019Published Date: May 17, 2019DOI: 10.23880/ipcm-16000160*Corresponding author: Dr. Palaksha MN, Professor& Head, Department of pharmacology, Bharathi College ofpharmacy, Bharathinagara, Mandya dist. Karnataka 571422, India, Tel: 9293785255; Email: palaksha.mn@gmail.comAbstractThe present study was carried out to evaluate the qualitative analysis of various phytochemicals and in-vitro evaluationof antibacterial and anthelmintic activities of Ethanolic leaf extracts of Sauropus androgynus (Phyllanthaceae). Theextracts were subjected to preliminary phytochemical screening for the identification of different phytoconstituents, theleaves indicated the presence of proteins, resins, steroids, tannins, glycosides, reducing sugar, carbohydrates, saponins,sterols, terpenoids, acidic compounds, cardiac glycosides, catechol, phenols, alkaloids, flavanoids. The differentconcentrations of Ethanolic extracts were subjected to antibacterial screening against various Gram positive and Gramnegative bacteria and the results were compared with that of standard drug Gentamycin. Different concentrations of leafextracts were tested to evaluate Anthelmintic activity using Sauropus androgynus (Indian earthworm). The extracts wasfound to possess vermifuge and vermicidal activity and the results were compared with standard drug Albendazole.Keywords: Phytochemical Screening; Sauropus Androgynus; Antibacterial Activity; Sauropus Androgynus; AnthelminticActivity; Gentamycin; AlbendazoleIntroductionSince the ancient time naturally occurring plants haveplayed an important role in the discovery of newtherapeutic agents. Almost all antibiotics are subjected tothe problem of bacterial resistance. Therefore, newerherbal antibacterial compounds from plants to overcomethe resistance are under investigation [1]. A wide range ofmedicinal plant parts is used for extract as raw drugs andthey possess varied medicinal properties [2] Sauropusandrogynus L. Merr., also known as katuk, star gooseberry,or sweet leaf, is a shrub grown in some tropical regions asa leaf vegetable. It is most popular in South Asia andSoutheast Asia. In India it also known as MultivitaminPlant as it contains an excellent source of vitamins A,B, Ccarotenoid and also it has high nutritive value and containsphytochemicals which can act as antioxidant [3]. The leafyvegetable Sauropus androgynus is commonlyused as anEvaluation of In-vitro Antibacterial and Anthelmintic Activities of SauropusAndrogynus (Phyllanthaceae) Plant ExtractsInt J Pharmacogn Chinese Med

International Journal of Pharmacognosy and Chinese Medicine2effective medicinal herb in the treatment of diabetics,cancer, inflammation, microbial infection, cholesterol andallergy due to its antioxidant effect [4].Materials and MethodsPlant Material Collection and Preparation ofExtractThe leaves of Sauropus androgynus were collected nearBharathinagar, Mandya district, Karnataka, India. Theleaves were shade dried, powdered and sieved through40mesh. The powdered leaves were successively extractedby maceration process using 90% ethanol as solvent. Theextracts obtained were evaporated under drying toremove the solvent completely a paste form of drug isobtained these were taken for further studies.Phytochemical ScreeningPhytochemical analysis was performed using standardprocedures: The freshly prepared extracts of Sauropusandrogynus was qualitatively tested for the presence ofchemical constituents. Phytochemical screening of theextracts was performed using standard procedures [5,6].Test for Carbohydrates by Molisch’s TestTo the test solution add few drops of alcoholic anaphthol, then add few drops of concentrated sulphuricacid through sides of test tube, purple to violet colour ringsat the junction of two layersTest for Reducing Sugar by Fehling’s Test: About 0.5geach portion was dissolved in distilled water and filtered.The filtrate was heated with 5mlof equal volumes ofFehling’s solution A and B. formation of a red precipitate ofcuprous oxide was an indication of the presence ofreducing sugars.Test for Protein by Xanthoprotein Test: To the 5ml oftest solution, add 1ml of concentrated nitric acid and boil,yellow precipitate is formed. After cooling it, add 40%Sodium hydroxide solution orange color is formed.Test for Terpenoids by Salkowski Test: Treat the extractwith few drops of concentrated sulphuric acid, red color atlower layer indicates presence of steroids and formation ofyellow colored lower layer indicates presence oftriterpenoids.Test for Alkaloids by Mayer’s Reagent: Alkaloids givecream color precipitate with Mayer’s reagent (Potassiummercuric iodide solution)Palaksha MN, et al. Evaluation of In-vitro Antibacterial andAnthelmintic Activities of Sauropus androgynus (Phyllanthaceae) PlantExtracts. Int J Pharmacogn Chinese Med 2019, 3(2): 000160.Test for Glycosides by Borntrager’s Test: Boil the testmaterial with 1ml sulphuric acid in a test tube for fiveminutes. Filter while hot. Cool the filtrate and shake withequal volume of dichlomethane or chloroform. Separatethe lower layer of dichlomethane or chloroform and shakeit with half of its volume of dilute ammonia. A rose pink tored color is produced in the ammonical layer.Test for Cardiac Glycosides by Keller-Killian Test:Extract the drug with chloroform and evaporate it todryness. Add 0.4ml of glacial acetic acid containing traceamount of ferric chloride transfer to a small test tube, addcarefully 0.5ml of concentrated sulphuric acid by the sideof the test tube. Acetic acid layer shows blue color.Antibacterial ActivityTest organism usedThe various organisms like Staphylococcus aureus,Pseudomonas aeruginosa, Bacillus subtilis, Escherichia coliare procured from Department of Microbiology BharathiCollege of pharmacy Bharathinagara, Mandya district,Karnataka, IndiaAntibacterial AssayLeaf extracts of Sauropus androgynus was evaluated forantibacterial activity against several gram positive andgram negative organisms. The antibacterial activity of Leafextracts of Sauropus androgynus was performed usingAgar cup-plate method 20ml of sterile nutrient agarmedium was poured into sterile Petri-dishes and allowedto solidify [7]. The Petri dishes were incubated at 370C for24 hours to check for sterility. The medium was mixedwith the organisms by pour plate method using sterile agarbroth (4ml) contained 1ml culture. Bores were made onthe medium using sterile borer. Leaf extracts of Sauropusandrogynus was dissolved in water to obtained differentconcentration (100, 200 mg/ml) and sterilized by filtrationthrough a whattman filter paper no. 1, and 0.5 ml of thedifferent concentration of extraction were added to therespective bores. 0.5 ml of Gentamycin at a concentrationof (40 µg/ml) was taken as standard reference. All theplates were kept in a incubator at 37 0C for 24 hours. Thepresence of definite zone of inhibition of any size aroundthe cup indicated antibacterial activity. The diameter of thezone of inhibition was measured and recorded.Anthelmintic minthic assay was performed on healthy adultIndian earth-worms Pheritima posthuma due to itsanatomical and physiological resemblance with theCopyright Palaksha MN, et al.

International Journal of Pharmacognosy and Chinese Medicine3intestinal roundworm parasites of human beings wereused in the present study. Because of easy availability.Earth worms have been used widely for the evaluation ofAnthelmintic compounds. All the earth worms were ofapproximately equal size (6cm). They were collected fromlocal moist place of Bharathi College of pharmacy situatedin Bharathinagara, Mandya district.Anthelmintic AssayAnthelmintic activity was evaluated by exposing theadult Pheritima posthuma to different concentration Leafextracts of Sauropus androgynus. The anthelmintic activitywas performed according to the method of Ghosh et al. [8]with slight modifications. The ethanolic extracts ofSauropus androgynus were dissolved in minimum amountof water and then volume was adjusted. 10 ml offormulation containing two different concentrations ofeach of the extract of extracts were prepared and 6 worms(same type) were placed in the petri dishes. The standardSl.no1234567drug and extract solutions were prepared freshly beforestarting the experiment. Time for paralysis was notedwhen no movement could be observed except when theworms were shaken vigorously. Time for death of wormswere recorded after sometime that worms neither movedwhen shaken vigorously nor when dipped in hot water(50 C) followed by fading away of their body colours.Albendazole (20mg/ml) was used as reference standard.ResultsPreliminary Phytochemical Screening of Sauropusandrogynus leaves indicated the presence of proteins,resins, steroids, tannins, glycosides, reducing sugar,carbohydrates, saponins, sterols, terpenoids, acidiccompounds, cardiac glycosides, catechol, phenols, alkaloids,flavonoids. The results of the phytochemical screening aregiven in the following (Table 1).Phytochemical analysis of SauropusandrogynusName of the testTest for carbohydratesMolisch testTest for reducing sugarFehling’s testTest for ProteinsXanthoprotein testTest for alkaloidsMayer’s testTest for Cardiac glycosidesKeller killian’s testTest for glycosidesBorntrager’s testTest for terpenoidsSalkowski test Table 1: Phytochemical constituents analysis of Sauropus androgynus. Prescence, - AbsenceAnti-Bacterial ActivityThe leaf extracts of Sauropus androgynus was studiedfor antibacterial activity employing standard cylindermethod. Microbes used were Bacillus a,Escherichia coli. Both gram-positive and gram-negativePalaksha MN, et al. Evaluation of In-vitro Antibacterial andAnthelmintic Activities of Sauropus androgynus (Phyllanthaceae) PlantExtracts. Int J Pharmacogn Chinese Med 2019, 3(2): 000160.bacteria were sensitive to the extract (Table 2, Figure 1).The antibacterial activity of the leaf extracts of Sauropusandrogynus was related to their chemical composition. Thediameters of the inhibition zones were measured inmillimeter [9].Copyright Palaksha MN, et al.