International Journal of Pharmaceutical Applications ISSN 0976-2639. Vol 2, Issue 1, 2011, pp 105-114 http://www.bipublication.com

EVALUATION OF ANTIMICROBIAL ACTIVITY OF GLUTINOSA

Poornima .V. Hosamath, Department of Biotechnology, Basaveshwar Science College, Bagalkot. Karnataka.

Abstract

Litsea glutinosa is widely available throughout . Litsea glutinosa is called as Indian Laurel in English and as Medhasaka in Sanskrit belonging to the family . The powdered material of Litsea glutinosa bark was extracted separately by continuous hot extraction process using soxhlet apparatus with different solvents in increasing order of polarity from Petroleum ether, Ethanol, to finally water. After extraction the extracts were subjected to Lyophilization to get dry extract & preserved in aseptic condition. The dried extracts were subjected to various phytochemical analysis to detect the presence of different phytoconstituents like alkaloids, glycosides, flavonoids, saponins, tannins, phenolic compounds. The petroleum ether extract, ethanolic extract and aqueous extracts of the Litsea glutinosa bark have the antibacterial & antifungal activity against Gram positive Staphylococcus aureus bacteria using reference standard like Procain pencillin,Gram negative like Pseudomonas aeruginosa, Salmonella typhi, E-coli bacteria using reference standard like Streptomycine sulphate. And fungal species like Aspergillus fumigatus and Candida albicans using reference standard like Griseofulvin. The bark of Litsea glutinosa, “ is one of the most popular of native drugs”, is considered to be capable of relieving pain, arousing sexual power & good for stomach are considered to be mildly astringent, include treatment of diarrohea & dysentery. Litsea glutinosa is widely used as a demulcent & as an emollient. Petroleum ether extract & Ethanolic extracts showed good activity against Pseudomonas aeruginosa. Aqueous extract & Petroleum ether extracts showed less activity as that of Ethanolic extract against Salmonella typhi. Then Ethanolic extract showed more effective activity against E-coli. Ethanolic extract had very high activity against Staphylococcus aureus. Likewise Petroleum ether extract of Litsea glutinosa effective in inhibiting the growth of Aspergillus fumigatus & Candida albicans. The phytochemical constituents of bark of Litsea glutinosa showed the effective antibacterial & antifungal activity.

Key words : Litsea glutinosa, Bark extracts, Antibacterial activity, Antifungal activity, Phytochemicals, Pathogens.

Introduction

Traditional knowledge regarding of cost of the drug, to overcome this problem medicinal and their use by indigenous plants growing around us are utilized without cultures are not only useful for conservation scientific validation. of cultural traditions and biodiversity but also for community healthcare and drug The use of higher plants and their extracts to development in the present and future. Due to treat infections is an age-old practice. the poor sanitary conditions, Sunburns Traditional medicinal practice has been Psoriasis and to the climatic conditions the known for centuries in many parts of the infections of the skin are common in the rural world. Ayurveda, the science of life, areas. Therapy with synthetic tropical prevention and longevity is the oldest and applications have most side effects and most holistic medical system available on the cannot be afford by the people due to import EVALUATION OF ANTIMICROBIAL ACTIVITY OF LITSEA GLUTINOSA

planet today. Herbal medicines are gaining So we have used the Petroleum ether, interest because of their cost effective and Ethanolic, Aqueous extracts of the Litsea eco-friendly attributes[3]. glutinosa bark, these all extracts have antibacterial and antifungal activity against For a long period of time, plants have been a gram-positive S.aureus bacteria using valuable source of natural products for reference standard like Procaine penicillin, maintaining human health, especially in the gram-negative like P.aeruginosa, Salmonella last decade, with more intensive studies for thphi, E-coli bacteria using reference natural therapies. According to World Health standard like Streptomycine sulphate and [4] Organization , medicinal plants would be fungal species Aspergillus fumigatus. Cndida the best source to obtain a variety of drugs. albicans using reference standard like About 80% of individuals from developed Griseofulvin. countries use traditional medicine, which has compounds derived from medicinal plants. MATERIALS AND METHODS : Therefore, such plants should be investigated I. Preliminary phytochemical screening: to better understand their properties, safety Phytochemistry is the chemistry dealing with and efficiency[1]. plants or products or natural products. The natural products comprise different The use of plant extracts and phytochemicals, therapeutically active (alkaloids, glycosides, both with known antimicrobial properties, flavonoids etc.) or inactive (starch, cellulose can be of great significance in therapeutic etc.) chemical constituents. The extract was treatments. In the last few years, a number of subjected to preliminary qualitative chemical studies have been conducted in different tests[7][ 9]. countries to prove such [6][8][12)[2][14][11][10] efficiency . Many plants have Carbohydrates: been used because of their antimicrobial Carbohydrates are the most abundant class of properties, which are due to compounds organic compounds found in living synthesized in the secondary metabolism of organisms. Chemically, carbohydrates are plant. These products are known by their simple organic compounds that are aldehydes active substances, for example, the phenolic or ketones with many hydroxyl groups added compounds which are part of the essential and composed of carbon, hydrogen and [5] [13] oils , as well as in tannin . oxygen. The basic carbohydrate units are called monosaccharides and general Litsea glutinosa is widely available stoichiometric formula of an unmodified throughout India. The fresh dried stem barks monosaccharide is (CH2O)n i.e. hydrates of were collected from the forests of carbon. Carbohydrates are mainly grouped in Ananthanahalli near Harapanahalli, to two major classes as simple sugars Karnataka. The bark was subjected to coarse (saccharides) and polysaccharides[7][ 9]. powdered (#:44) to obtain uniform texture. Chemical tests: The sieved powder was stored in air tight & 1. Molisch’s Test (General test): The test is high density polyethylene containers before positive for soluble as well as insoluble extraction. The powdered material of Litsea carbohydrates. To 2-3 ml of aqueous glutinosa bark was extracted separately by extract add few drops of Molisch’s continuous hot extraction process using reagent (alpha-naphthol in alcohol) and soxhlet apparatus with different solvents in concentrated sulphuric acid along sides increasing order of polarity from petroleum of test tube – Voilet colour ring at the ether, ethanol & aqueous extracts were junction of two liquids. subjected to lyophilization to get dry extract 2. Test for reducing sugars & preserved in aseptic conditions. The dried i. Fehling’s Test: Mix 1ml of T.S.+ 1ml of extracts reconstituted in DMSo. Fehling’s solution A + 1ml of Fehling’s solution B and boil on water bath for 5-

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10 min – First a yellow, then brick red Glycosides: ppt is observed. Glycosides are the organic compounds from ii. Benedict’s Test: Mix equal volume of plants or animal sources which on enzymatic Benedict’s reagent and test solution in or acid hydrolysis gives one or more sugar test tubes and heat on boiling water bath moieties (glycone) along with non- sugar for 5 min. Solution appears green, yellow moieties (aglycone or genin). Chemically, or red depending on amount of reducing they are the acetals or sugar ethers formed by sugar present in test solution. interaction of hydroxyl group each of non- 3. Test for monosaccharides sugar and sugar moieties, with a loss of water i. Barfoed’s Test: Mix equal volume of molecule. The linkage between glycone and Barfoed’s reagent and test solution. Heat aglycone is called glycosidic linkage and on on boiling water bath for 1-2 min and the basis of this linkage, alpha and beta cool. Red colour ppt is observed. stereoisomers are assigned[7][ 9]. 4. Test for non- reducing sugars Chemical tests: i. Test solution does not give response to 1. Keller Killiani Test (for deoxysugars): To Fehling’s and Benedict’s tests. 2 ml of extract add few drops of glacial Alkaloids : acetic acid, 1 drop of 5% FeCl3 solution Alkaloids are heterogeneous group of natural and conc. H2SO4. Reddish brown color substances which are basic in nature and appears at the junction of two liquid contain one or more nitrogen atoms and layers and upper layer appears bluish possess specific physiological actions when green. used in small quantities. The qualitative 2. Legal’s Test (test for cardenoloids): To chemical tests used for detection of alkaloids aqueous or alcoholic extract, add 1 ml of are depends on characteristics of alkaloids to pyridine and 1ml of sodium nitroprusside give precipitates as salts of organic acids or solution. Pink to red color appears. with compounds of heavy metals like gold, 3. Modified Borntragger’s Test (for C- platinum, mercury etc. the different reagents glycosides): To 5 ml of extract, add 5 ml used are Mayer’s reagent (potassium- of 5% FeCl3 and 5 ml dil. HCl. Heat for 5 mercuric-iodide solution), Dragendorff’s min on boiling water bath. Cool and add reagent (potassium-bismuth-iodide solution), benzene or any organic solvent, shake Hager’s reagent (picric acid) and Wagner’s well. Separate organic slayer, to which reagent (iodine-potassium-iodide solution). add equal volume of dilute ammonia. Special tests are required for highly water Ammonical layer shows pinkish red soluble alkaloids like caffeine[7][ 9]. color.

Chemical tests: Flavonoids: Evaporate the alcoholic extract and to the The flavonoids are polyphenolic compounds residue add dilute HCl. Shake well and filter, possessing 15 carbon atoms; two benzene perform the following tests with filtrate. rings joined by a liner three carbon chain and 1. Mayer’s Test: 2-3 ml of filtrate + few are categorized, according to chemical drops of Mayer’s reagent – Creamy white structure, in to flavonols, flavones, ppt. flavanones, isoflavones, catechins, 2. Wagner’s Test: 2-3 ml of filtrate + few anthocyanidins and chalcones. The term drops of Wagner’s reagent – Reddish flavonoid (or bioflavonoid) refers to a class brown colored ppt. of plant secondary metabolites and are most 3. Dragendorffs’s Test: To 2-3 ml of filtrate commonly known for their antioxidant add few drops of Dragendorffs’s reagent- activity. Flavonoids are mostly yellow Orange red colored ppt. were obtained. colored substances and chemically they are 4. Hager’s Test: 2-3 ml of filtrate + few derivatives of phenyl benzo gama-pyrone drops of Hager’s reagent – Yellow ring[7][ 9]. colored ppt.

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Chemical tests: Phenolic compounds: 1. Shinoda Test (Mg/HCl): 2-3 ml of test Phenolic compounds are plant-based solution + pinch of Mg powder + 2-3 materials, phytochemicals which are drops of conc. HCl – Deep red or synthesized primarily from products of the magneta color. shikimic acid pathway, have several 2. Pew’s Test (Zn/HCl): Pinch of Zn important roles in plants. Tannins, lignans, powder + few ml of conc. HCl + 1-2 ml flavonoids, and some simple phenolic of test solution, which gives deep purple compounds serve as defenses against red color. herbivores and pathogens[7][ 9]. 3. Few ml of alcoholic extract + 1-2 drops 1. To 2 ml of test solution add 0.5 ml FeCl3 of conc. H2SO4 + magnesium turnings- solution – Formation of intense color. Deep cherry red color. 4. To 1 ml of aq NaOH add 1 ml of test II. Antibacterial activity: solution – Yellow color, which The compounds were tested in-vitro for their decolorizes after addition of acid. antibacterial activity against four 5. Addition of lead acetate solution to small microorganisms viz. Escherichia coli, quantities of residue gives yellow colored Pseudomonas aeruginosa, Staphylococcus ppt. aureus, Salmonella typhi which are pathogenic in human beings. Saponins: Specifically, they are amphipathic glycosides Method: grouped phenomenologically by the soap-like Cup-plate agar diffusion method using foaming they produced when shaken in Nutrient agar. aqueous solutions, and structurally by their being composed of one or more hydrophilic Materials used : glycoside moieties combined with a i. Nutrient broth (Himedia) lipophilic triterpene derivative[7][ 9]. ii. Nutrient agar (Himedia) 1. Foam Test: Extract is shaken vigorously iii. 18-24 hrs growth culture in nutrient with distilled water in test tubes agar Honey comb like foam is produced. iv. Sterile petridishes Tannins: v. Sterile micropipettes Tannins are astringent, bitter plant vi. Sterile cotton swabs polyphenols that either bind precipitate or vii. Sterile cork borer shrink proteins. They form colloidal solutions viii. Sterile test tubes with water and are non-crystalline Preparation of Nutrient broth: substances. i. Nutrient broth - 3.8 gm Tannins exhibits some chemical reactions, ii. Distilled water - 100 ml 1. Gelatin Test: Extract is dissolved in 2% Above components were dissolved in 100 ml gelatin + 10% NaCl – White precipitate. distilled water and pH was adjusted to 7.2. 2. Extract + alcoholic vanillin solution + This solution was sterilized by autoclaving at dilute HCl – Pink color. 15 psi. for 20 min. 3. Extract + Lime water – Brown color. 4. Tannins are precipitated by salts of Preparation of Inoculums: copper, tin and lead. One day prior to these testing, inoculations of 5. Tannins show color reactions with iron the above bacterial cultures were made in the salts. Ferric chloride gives bluish black or nutrient broth and incubated at 370C for 18 – bluish green color and potassium 24 hrs. ferricyanide with ammonia gives deep Preparation of medium (Nutrient agar): red color. i. Nutrient agar - 2.8 gm ii. Distilled water - 100 ml

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The agar was dissolved in to distilled water Above components were dissolved in and pH was adjusted to 7.4+0.2. It was distilled water (1000 ml) and pH was sterilized by autoclaving at 15psi for 20 min. adjusted to 5.5 – 6.0. This solution was sterilized by autoclaving at 1210C for 10 min. Preparation of test solutions: Each test compound (5 mg) was dissolved in Preparation of Inoculums: dimethyl sulfoxide (5 ml) to give stock One day prior to these testing, inoculations of solution of concentration 1000 mcg/ml. Then the above fungal cultures were made in the 0.1 ml of this solution was used for testing. Sabouraud-Dextrose medium and then Preparation of standard solution: incubated at 370C for 18 – 24 hrs. Standard drug Procain pencillin and Preparation of Sabouraud-Dextrose agar: Streptomycin sulphate were used. The i. Dextrose - 40 gm concentration was 100mcg/ml. ii. Neopeptone - 10 gm Method of testing: iii. Agar - 15 gm Nutrient agar plates were prepared by All these components were dissolved in pouring 15 – 20 ml of the medium in to each distilled water (1000 ml) and pH was sterilized petridish and were allowed to set at adjusted to 5.5 - 6.0. It was sterilized by room temperature. The cell suspension was autoclaving at 1210C for 10 min. standardized to the density of 530 nm using Preparation of test solutions: spectrophotometer and was inoculated over Each test compound (5 mg) was dissolved in the surface of agar medium using sterile dimethyl sulfoxide (5 ml) to give stock cotton swab. The four cups were scooped in solution of concentration 1000 mcg/ml. Then each plate using a sterile cork borer of 8 mm 0.1 ml of this solution was used for testing. diameter. Preparation of standard solution: Then the solutions of test compounds (0.1) Standard drug Griseofulvin was used. The were added in cups by using micropipettes concentration was 100mcg/ml. and these plates were incubated at 370C for Method of testing: 48 hrs. The zone of inhibition was measured Sabouraud-Dextrose agar plates were in mm for each organism. prepared by pouring 15 – 20 ml of the medium in to each sterilized petridish and III. Antifungal activity : were allowed to set at room temperature. The The compounds were tested in-vitro for their cell suspension was standardized to the antifungal activity against Candida albicans density of 530 nm using spectrophotometer and Aspergillus fumigatus. and was inoculated over the surface of agar Method: medium using sterile cotton swab. The four Cup-plate agar diffusion method using cups were scooped in each plate using a Sabouraud-Dextrose agar. sterile cork borer of 8 mm diameter, corresponding to control, standard and test Materials used : solution. i. Sabouraud-Dextrose agar The solutions of each test compounds (0.1) ii. 18-24 hrs growth culture on were added in cups by using micropipettes Sabouraud-Dextrose agar and these plates were incubated at 370C for iii. Sterile petridishes 48 hrs. The zone of inhibition was measured iv. Sterile micropipettes in mm for each organism. v. Sterile cotton swabs RESULTS: vi. Sterile cork borer (8 mm) Phytochemical screening of Litsea vii. Sterile test tubes glutinosa bark extract: Preparation of Sabouraud-Dextrose medium: The Litsea glutinosa bark extract was i. Dextrose - 40 gm subjected to preliminary qualitative chemical ii. Neopeptone - 10 gm tests. These tests were conducted to know the constituents, present in the plant extract. The

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Litsea glutinosa bark extract has showed the + Presence of constituents presence of alkaloids in Aqueous extract, - Absence of constituents Ethanolic exract, and Petroleum ether extract. Antibacterial activity: Sterols were absent in all the three kinds of Petroleum ether extract, Ethanolic extract, extracts. Glycosides & Carbohydrates were aqueous extract of Litsea glutinosa has present in all the extract of plant. Mucilage showed the antibacterial activity. was absent in Aqueous extract, Ethanolic Pseudomonas aeruginosa has showed more extract as well as Petroleum ether extract. activity in the Petroleum ether extract & The presence of Starch was identified by Ethanolic extract, least activity in aqueous iodine test-only the Aqueous extract has extract. Salmonella typhi has showed good showed presence of starch but not the activity against Ehanolic extract, moderate Ethanolic & Petroleum ether extract. Other activity in aqueous extract & poor activity test for the detection of starch was Tannic against Petroleum ether extract. In E-coli acid test, presence of tannic acid in all the significant activity was observed in Ehanolic extracts of plant has indicated the presence of extract, good activity against aqueous extract starch. Proteins, Cardiac glycosides, & least activity in Petroleum ether extract. Saponins, Tannins & Phenolics were present DMSo was referred as blank showed in all the three extracts of Litsea glutinosa, minimum activity & Strptomycin sulphate as but the flavonoids were absent in the extracts. standard has maximum activity. Finally the phytochemical screening of Litsea Staphylococcus aerus, the gram+ve bacteria glutinosa bark extract constituents were has showed maximum activity against Tannin, beta-sitosterols and Boldine, nor- Ethanolic extract, very good activity in boldine, laurotetanine, N- Aqueous extract & has least activity in methylactinodaphnine, quercetine, sebiferine, Petroleum ether extract. DMSo referred as litsiferine. Kaempferol-3-glucoside, blank has minimum activity & Procaine aminoacids, quercetine-3-rhamnoside, penicillin as standard has maximum activity. Kaempferol-7-aminoglucoside, So all the extracts of Litsea glutinosa has Pelargonidine-5-glucoside, naringenin-7- antibacterial activity of both gram+ve and monorhamnoside, mono and sesquterpines, gram-ve bacteria. beta-amirine acetate. Diameter of mean zone Sl. Aqueous Ethanolic Petroleum of inhibition(mm) after Constituents No extract extract ether extract 24 to 48 hours Drugs 1 Alkoloides + + + Pseudomonas 2 Sterols - - - aeruginosa 3 Glycosides + + + 50mg/ml 100mg/ml Blank 16 16 4 Carbohydrates + + + Petroleum ether 5 Mucilage - - - 17 19 extract 6 Starch Ethanolic extract 17 19 a. Iodine test + - - Aqueous extract 16 17 b.Tannic acid test + + + Streptomycin 29 29 7 Proteins + + + sulphate 8 Cariac glycosides + + + Table 2: Effect of petroleum ether, ethanolic and 9 Saponins + + + aqueous extracts of Litsea glutinosa bark on 10 Tannins and + + + Pseudomonas aeruginosa. phenolics 11 Flavonoides - - -

Table 1 : Phytochemical screening of Litsea glutinosa bark extracts

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Note : 1. Average of three plates have taken for Diameter of mean each extract zone of inhibition(mm) 2. Diameter of the cup = 8 mm Drugs after 24 to 48 hours 3. The values shown in parenthesis Salmonella typhi indicates activity index. 50mg/ml 100mg/ml 4. Activity index more than 0.5 is Blank 12 12 considered as significant activity.Hence Petroleum ether 11 12 Petroleum ether, Ethanolic extracts extract shows significant antimicrobial activity. Ethanolic extract 13 17

Aqueous extract 11 14

Streptomycin 24 24 29 sulphate 30 29

50 mg/ml 25 Table 3: Effect of petroleum ether, ethanolic and 100 mg/ml aqueous extracts of Litsea glutinosa bark on Salmonella 19 typhi. 20 19 17 17 17 16 16 16 Diameter of mean zone 15 of inhibition(mm) after Drugs 24 to 48 hours 10 Escherichia coli zone inhibition of of Diameter 5 50mg/ml 100mg/ml

Blank 13 13 0 Blank Petroleum Ether Ethanolic Aqueous Streptomycin Petroleum ether Extract Extract Extract Sulphate 14 17 D rugs extract Ethanolic extract 20 24 Fig.1 : Effect of petroleum ether, ethanol, aqueous extracts of Litsea glutinosa bark on Pseudomonas Aqueous extract 14 18 aeruginosa Streptomycin 30 30 30 30 sulphate 30 Table 4: Effect of petroleum ether, ethanolic and 50 mg/ml 25 24 aqueous extracts of Litsea glutinosa bark on 100 mg/ml Escherichia coli. 20 20 18 Diameter of mean zone 17

of inhibition(mm) after 14 14 15 13 13 Drugs 24 to 48 hours Staphylococcus aureus 50mg/ml 100mg/ml 10 Blank 13 13 of zoneinhibition of Diameter 5 Petroleum ether 13 15 extract 0 Ethanolic extract 17 24 Blank Petroleum Ether Ethanolic Aqueous Streptomycin Extract Extract Extract Sulphate Aqueous extract 15 19 Drugs

Procaine penicillin 28 28 Fig.2 : Effect of petroleum ether, ethanol, aqueous extracts of Litsea glutinosa bark on Escherichia Table 5: Effect of petroleum ether, ethanolic and coli aqueous extracts of Litsea glutinosa bark on Staphylococcus aureus .

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with minimum activity & standard, 25 24 24 Griseofulvin has activity within range. All the bacterial species have more activity 50 mg/ml 20 100 mg/ml against Ethanolic extract as compared to 17 aqueous extract & Petroleum ether extract. Likewise all fungal species have showed 15 14 13 effective activity against Petroleum ether 12 12 12 11 11 extract, but not in the Ehanolic extract &

10 Aqueous extract.

Diameter of zone of inhibition zone of of Diameter Diameter of mean 5 zone of inhibition(mm) after

0 Drugs 24 to 48 hours Blank Petroleum Ether Ethanolic Aqueous Streptomycin Aspergillus fumigatus Extract Extract Extract Sulphate Drugs 50mg/ml 100mg/m Fig.3 : Effect of petroleum ether, l ethanol, aqueous extracts of Litsea glutinosa bark Blank 15 15 on Salmonella typhi Petroleum ether 17 18 extract 30 28 28 Ethanolic extract 13 14

50 mg/ml Aqueous extract 13 14 25 24 100 mg/ml Griseofulvin 17 17

19 20 Table 6: Effect of petroleum ether, ethanolic and 17 aqueous extracts of Litsea glutinosa bark on 15 15 Aspergillus fumigatus. 15 13 13 13

10 Diameter of mean zone of Diameter of zone of inhibition of zone of Diameter

5 inhibition(mm) after Drugs 24 to 48 hours

0 Candida albicans Blank Petroleum Ether Ethanolic Aqueous Streptomycin 50mg/ml 100mg/ml Extract Extract Extract Sulphate Drugs Blank 14 14 Petroleum ether 19 23 Fig.4 : Effect of petroleum ether, ethanol, aqueous extract extracts of Litsea glutinosa bark on Ethanolic extract 13 15 Staphylococcus aureus Antifungal activity: Aqueous extract 12 15 Aspergillus fumigatus has showed the more Griseofulvin 15 15 activity against Petroleum ether extract, less activity in Ethanolic extract and poor activity Table 7: Effect of petroleum ether, ethanolic and in Aqueous extract. DMSo referred as blank aqueous extracts of Litsea glutinosa bark on & it has showed considerable activity. Candida albicans. Griseofulvin as standard has showed average activity. Similarly Candida albicans also has Note : the maximum activity against Petroleum 1. Average of three plates have taken for ether extract and moderate activity in each extract Ethanolic extract, least activity in aqueous 2. Diameter of the cup = 8 mm extract. Here also DMSo referred as blank

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3. The values shown in parenthesis Mucilaginous bark is used in diarrhea & indicates activity index. dysentery treatment and also for sprains, 4. Activity index more than 0.5 is bruises & rheumatic gouty joints. considered as significant activity.Hence Petroleum ether, Ethanolic extracts Petroleum ether & Ethanolic extract showed shows significant antimicrobial activity. good activity as compare to the Aqueous extract against Pseudomonas aeruginosa. 18 18 50 mg/ml 17 17 17 Aqueous extract & Petroleum ether extracts 100 mg/ml showed less activity as that of Ethanolic 16 15 15 14 14 extract against Salmonella typhi. Then 14 13 13 Ethanolic extract showed more effective activity against E-coli but the Aqueous 12 extract & Petroleum ether extract showed 10 moderate activity against E-coli. Ethanolic extract had very high activity, aqueous 8 extract moderate & Petroleum ether extract 6 showed less activity against Staphylococcus aureus. Diameter of zone of of inhibition of zone Diameter 4 So all extracts of Litsea glutinosa inhibited 2 both gram-positive & gram-negative bacteria. 0 Likewise Petroleum ether extract of Litsea Blank Petroleum Ether Ethanolic Aqueous Streptomycin Extract Extract Extract Sulphate glutinosa showed more effective activity Drugs against Aspergillus fumigatus & Candida Fig.5 : Effect of petroleum ether, ethanol, aqueous albicans as that of Ethanolic & Aqueous extracts of Litsea glutinosa bark on Aspergillus extract of Litsea glutinosa. So the Petroleum fumigatus ether extract of Litsea glutinosa inhibited the growth of fungal species.

25 23 Tannin, beta-sitosterols and Boldine, nor- 50 mg/ml 100 mg/ml boldine, laurotetanine, N- 20 19 methylactinodaphnine, quercetine, sebiferine, 15 15 litsiferine. Kaempferol-3- 15 15 15 14 14 13 glucoside,aminoacids, quercetine-3- 12 rhamnoside, Kaempferol-7-aminoglucoside, 10 Pelargonidine-5-glucoside, naringenin-7- monorhamnoside, mono and sesquterpines, [44] Diameter of zone inhibition zone of of Diameter 5 beta-amirine acetate . These are the phytochemical constituents of bark of Litsea glutinosa, so it had showed the effective 0 Blank Petroleum Ether Ethanolic Aqueous Streptomycin results of antibacterial & antifungal activity. Extract Extract Extract Sulphate Drugs Fig.6 : Effect of petroleum ether, ethanol, aqueous Conclusion extracts of Litsea glutinosa bark on Candida In this study Litsea glutinosa bark extracts albicans showed the effective antimicrobial activity.After knowing the constituents of DISCUSSION: bark extracts, treated with bacterial & fungal Litsea glutinosa is one of the most popular of species. Pseudomonas aeruginosa has native drugs. The bark of Litsea glutinosa is showed more activity in Petroleum ether widely used as a demulcent. Ground & pasted extract & Ethanolic extract, least activity in material is used as an emollient. aqueous extract. Salmonella typhi has showed good activity against Ehanolic extract,

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moderate activity in aqueous extract & poor 3. Ellof, J.N. Which extractant should be used activity against Petroleum ether extract. In E- for the screening and isolation of coli significant activity was observed in antimicrobial components from plants? J. Ehanolic extract , good activity against Ethnopharmacol. 60, 1-6, 1998. Aqueous extract & least activity in Petroleum 4. Fabida Barbieri Holetz, Greisiele Lorena Pessini, Neviton Rogerio Sanches, Diogenes ether extract. DMSo was referred as blank Aparicio Garcia Cortez, Celso Vataru showed minimum activity & Strptomycin Nakamura, Bendito Prado Dias Filho. sulphate as standard has maximum activity. Screening of some plants used in the Staphylococcus aerus, the gram+ve bacteria Brazilian Falk medicine for the treatment of has showed maximum activity against infectious diseases. Vol. 97(7): 1027-1031, Ethanolic extract, very good activity in October 2002. Aqueous extract & has least activity in 5. Jhon J. Rojas, Veronica J. Ochoa, Saul A. Petroleum ether extract. DMSo referred as Ocampo and John F. Munoz. Screening for blank has minimum activity & Procaine antimicrobial activity of ten medicinal plants penicillin as standard has maximum activity. use in Colombian folkloric medicine: A possible alternative in the treatment of non- Aspergillus fumigatus and Candida albicans nosocomial infection. 17 Feb. 2006. both has showed the maximum activity 6. Jonathan E. Kelmanson, Anna K. Jager and against Petroleum ether extract & least Johannes Van Staden. Zulu medicinal plants activity in Ethanolic extract as well as with antibacterial activity. 14 August 1999. Aqueous extract. Here also DMSo referred as 7. Jonsen, A.M.; Cheffer, J.J.C.; Svendsen, A.B. blank showed considerable activity and Antimicrobial activity of essential oils: a Griseofulvin as standard has minimum 1976-1986 literature review. Aspects of test activity.These results justify that, all the methods. Planta Med. 40, 395-398, 1987. extracts of Litsea glutinosa has antibacterial 8. L.M. Bedoya, S. Sanchez Palomino, M.J. activity of both gram+ve and gram-ve Abad, P. Bermejo and J. Alcami. Anti-HIV activity of medicinal plant extracts. 12 May bacteria and also has effective antifungal 2001. activity. 9. Mehlika Benli, Umit Bingol, Fatmagul Geven, Kerim Guney and Mazife Yigit, Ankara, Kastamonu, Kirikkale Universities of Acknowledgement Turkey. An investigation on the antimicrobial The author is thankful to Pricipal & Sri activity of some endemic plant species from V.M.Chandrashekar Assistant Professor of Turkey. 6 Nov. 2007. H.S.K Pharmacy College, Bagalkot. Pricipal 10. Ki-Bong Oh, Ji Hye Lee, Kyung-Mi & Faculty members of Botany department of Yoon, Soon-Chun Chung, Heung Bae Jeon, Basaveshwar Science college, Bagalkot. Jongheon Shin and Hyi-Seung Lee. Synthesis and antimicrobial activity of halogenated bis Sri.V.G.Jadimath Lecturer Govt PUcollege (hydroxyphenyl) methanes. 24 Nov 2008. Bagalkot for providing research facilities, 11. Osman Sag Dic and Musa Ozcan. guidance & encouragement. Antibacterial activity of Turkish spice References hydrosols. 25 June 2002. 1. Amal G, Al-Bakri and Fatma U. Afifi. 12. Phillipp. J. Sci. C6: 321-1911 University of Jordan. Evaluation of 13. Seenivasan Prabuseenivasan, Manickkam antimicrobial activity of selected plant Jayakumar and Savarimuthu Ignacimuthu. In extracts by rapid XTT calorimetry and vitro antibacterial activity of some plant bacterial enumeration. 31 May 2006. essential oils. 30 Nov 2006. 2. Anmara Hassan, Salma Rahman, Farah 14. Semma J. Patel, Nithin Venugopalan, S. Deeba and Shahid Mahmud. Applied Pradeep, G.M. Institute of Technology, chemistry research center, PCSIR Labs, Davangere, India. Screening for antimicrobial Complex, Ferozpur Road, Lahore, Pakistan. activity of weeds. The Internet Journal of Antimicrobial activity of some plant extracts Microbiology. 1 Nov 2007. Vol 4. having hepatoprotective effects. 19 Dec. 2008.

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