India correlates Daruharidra to Berberis aristata DC of Key words: Antimicrobial activity, Daruharidra, eye infection, methanolic stem extract. The highest antimicrobial activity of B. aristata root extract was found against S. aureus ( mm) in acetonic extract with an MIC of mg mL-1 and that of leaf . The antimicrobial activity of hydroalcoholic extracts of four Berberis species viz. Berberis aristata, Berberis asiatica, Berberis chitria and Berberis lycium were.
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Berberis aristata Berberidaceae is an important medicinal plant and found in the different region of the world. It has significant medicinal value in the traditional Indian and Chinese system of medicine. The aim of the present investigation was undertaken to find aristat the phytochemical presence and Antimicrobial activity of aqueous and alcholic extract of Berberis aristata. Present study includes determination of phytochemical analysis, antimicrobial study and estimation of total flavonoid content.
Preliminary phytochemical analysis showed the presence of carbohydrate, glycoside, alkaloid and flavonoid. Total flavonoid content was found to be 0.
Antimicrobial activity shows good antimicrobiwl result with gram positive bacteria. Berberis aristata belongs to the family Berberidaceae, is an important medicinal plant. The plant is an important medicine for the treatment of oxidative stress, remittent fevers, and is used as a cooling laxative to children and as a tonic medicine for liver and heart. The plant contains berberine, oxycanthine, epiberberine, palmatine, dehydrocaroline, jatrorhizine and columbamine, karachine, Four alkaloids, pakistanine, 1-Omethylpakistanine, pseudopalmatine chloride and pseudoberberine chloride[4,6].
It has hypotensive, immuno-stimulating, antiinflammatory, antimicrobial, antiprotozoal, activities. It has also antifungal, tuberculostatic antihelminthic, properties.
The Bacteria related, parasitic intestinal infections diarrhea, and ocular infections are the most well-known clinical uses of berberine.
It has been reported that berberine exhibits local anesthetic, enzyme inhibitory and antipyretic activities [7 — 10]. Berberis extracts and decoctions have showed significant antimicrobial activity against a variety of organisms including bacteria, viruses, fungi, protozoans, helminths, and Chlamydia.
Currently, the predominant clinical uses of berberine include bacterial diarrhea, intestinal parasite infections, and ocular trachoma infections. The most active ingredient of the plant is berberine, a quaternary isoquinoline alkaloid and the content of berberine is used for monitoring the quality of the plant. It is typically found in the roots, rhizomes and stem bark. Collection of Plant material and identification: Berberis aristata plant material root was collected from the panchmari at Madhya Pradesh India.
Identification of plant material was done in the Scan Research Laboratory Indripuri Bhopal and placed in herbarium and was well documented. Both aqueous and alcoholic extracts were prepared as described by  with slight modifications as adopted in previous studies . Fresh roots of B. The sterilized materials were grounded with a sterile pestle and mortar in sterile distilled water ml.
The homogenized tissue was centrifuged at 7, rpm for 15 min, supernatant was filter-sterilized and used as the aqueous extract. The supernatant was put in a hot water bath at 60 0C to evaporate the organic solution. The extracts were filtersterilized before use. Khandelwal, ,Kokate, and Tiwari et al.
The extracts obtained by solvent extraction were subjected to various qualitative tests detect the presence of plant constituents Carbohydrates, Alkaloids, Saponins, Tannins and Triterpenoids etc. The filtrate was shaken with 5ml of chloroform. The chloroform layer was separated in a porcelein dish the solvent is removed by gentle evaporation. This solution was carefully transferred to the surface of 2ml of concentrated sulphuric acid. A reddish brown layer is formed at the junction of two liquids and the upper layer slowly becomes bluish green, darkening with standing.
Extracts were treated with few drops of sodium hydroxide solution. Formation of intense yellow colour, which becomes colourless on addition of dilute ariztata, indicates the presence of flavonoid. Small quantity of alcoholic and aqueous extract was taken separately and 20ml of distilled water was added and shaken in graduated cylinder for 15 minutes length wise.
A 1cm layer of foam indicates the presence of saponins. No saponins were indicated in extract. The mixture was then heated on a water bath for hours. The formation of soap or partial neutralization of alkali indicates the presence of fixed oils and fats. But there was no such formation of soap which indicates the absence of fats.
Two or three granules of tin metal were dissolved in 2 ml of thionyl chloride solution. Then 1ml of the extract was poured into test tube and warmed, pink color was produced which indicates the presence of triterpenoids.
No such colour is produced which shows the absence of triterpenoids. Purple color is formed at the junction of the two liquids which reveals the presence of carbohydrates. If Pinkish color is formed shows presence of proteins but no such colour is formed indicating absence of proteins. Determination of total flavonoids content was based on aluminium chloride method. Disk diffusion method was used axtivity the determination of antimicrobial activity.
Clinical isolates of Gram-positive and Gram-negative bacteria were collected from the Scan Research laboratory Indripuri Bhopal for anti microbial activity assay. The clinical bacterial species used were Staphylococcus aureus, Anttimicrobial subtilis, Escherichia coli, Salmonella typhimurium. Discs containing aqueous beeberis alcoholic extracts at concentrations of 50, 25 and Ciprofloxacin was used as standard antibiotics.
Preliminary phytochemical analysis of Berberis aristata showed the presence of carbohydrate, glycoside, alkaloid, flavonoid while protein, saponin, triterpenoids and fats and fixed oils were absent.
Total flavonoid content was determined and found to be 0. The different three concentrations Different methods were followed to find out qualitatively the presence of phytochemical constituents in the extract. The extract of the plant possesed alkaloids, glycosides,flavonoids and carbohydrates while sasponins, fats and fixed oils, triterpenoids and proteins were absent.
The flavonoids in the presence of aluminum chloride have an deep yellow fluorescence which observed under UV spectrophotometer at nm. Estimation Total flavonoids content TFC. Alcoholic and aqueous extract showed antimicrobial activity against four tested bacteria.
Positive controls formed considerably sized inhibition zones against tested bacteria. Barberis aristata showed highest zone of inhibition for Bacillus subtilis followed by Staphylococcus aureus, Escherichia coli and Salmonella typhimurium. The antibacterial activity of the extract against clinical isolates was comparable to that of standard strains Table 3.
It is interesting to note down that the Gram-negative Bacteria reported here as prone to the extracts of B. Inhibition zones mm of aqueous and alcoholic extract of Berberis aristata against Gram-positive and Gramnegative bacteria species.
Physicochemical standards are usually used for determining the character, purity and strength of the drug source. These characters are also used to confirm the actual nature of the crude drug. Hence it plays an important role in preventing the possible steps of adulteration. The photochemical analysis conducted on Berberis aristata extracts revealed the presence of alkaloid, carbohydrate, glycoside, flavonoid and other phytoconstituents.
The phytochemical analysis revealed the presence of different phytoconstituents including triterpenoidal, phenolic and flavonoidal compounds. The medicinal properties of plants are due to the presence of different classes of secondary metabolites such as alkaloids, flavonoids, glycosides, phenols, saponins, sterols, etc .
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In the existing literature, berberine has been reported to be produced by various Berberis Spp. The most active constituent of Berberis aristata is berberine, a quaternary isoquinoine alkaloid and the content of berberine is used as biomaker of the plant. Flavonoids have a membrane permeability effect and are considered as potential antioxidants and have caring action against allergies, inflammation, platelet aggregation, microbes, ulcers and hepatoxins, [16 — 19].
Phenols and phenolic compounds are greatly used in skin infections, wound healing, inflammation; antioxidant, immune enhancers, anti clotting and hormone modulators .
Ethno botanically, Berberis aristata has been used against a broad range of ailments . Uneven activities of different plant parts root, stem of this plant have been reported against different bacterial and fungal human pathogens in different solvents [21 — 22].
A majority of the described antimicrobial effects of Berberis aristata have been accredited to their secondary metabolites, particularly alkaloid compounds . In this study, the extract of Berberis aristata are found to be most active in inhibiting the growth of pathogens viz. The root extract of Berberis aristata were found to be mainly active in inhibiting the growth of gram positive bacteria and gram negative bacteria.
The antimicrobial activity of Berberis aristata extract against tested bacterias may be due to the presence of secondary metabolites mainly berberine, an isoquinoine alkaloid with a bright yellow colour.
Published reviews have described a majority of antimicrobial effects of the extracts of Berberis species have been accredited to their secondary metabolites, especially alkaloids.
The present study clearly reveals the antimicrobial nature of Berberis aristata and recommended that this plant could be exploited in the management of diseases caused by microbes in plants and animals.
Keywords Antimicrobial, Berberis aristata, aqueous and alcoholic, phytochemical Introduction Berberis aristata belongs to the family Berberidaceae, is an important medicinal plant.
It is typically found in the roots, rhizomes and stem bark Materials and Methods Collection of Plant material and identification: Preparation of the Plant extract Both aqueous and alcoholic extracts were prepared as described by  with slight modifications as adopted in previous studies .
Aqueous extracts Fresh roots of B. Test for Flavonoids Extracts were treated with few drops of sodium hydroxide solution.
Formation of intense yellow colour, which becomes colourless on addition of dilute acid, indicates the presence of flavonoid Test of Saponins Small quantity of alcoholic and aqueous extract was taken separately and 20ml of distilled water was added and shaken in graduated cylinder for 15 minutes length wise.
Test of Fats and Fixed oils 1ml of the extract few drops of 0. Test for Triterpenoids Two or three granules of tin metal were dissolved in 2 ml of thionyl chloride solution.
Phytochemistry and Pharmacology of Berberis Species
Total flavonoids berheris estimation Determination of total flavonoids content was based on aluminium chloride method. Screening for antimicrobial activity Disk diffusion method was used for the determination of antimicrobial activity.
Statistical Analysis Data were statistically analyzed by using either one way or two way analysis of variance ANOVA Results Preliminary phytochemical analysis of Berberis aristata showed the presence of carbohydrate, glycoside, alkaloid, flavonoid while protein, saponin, triterpenoids and fats and fixed oils were absent.
Presence of Phytochemicals Different methods were followed to find out qualitatively the presence of phytochemical constituents in the extract. Absorbance 0 0 0 1 25 0. Blasko, karachine, an usual protoberberine alkaloid,Journal of American chemical societyChemical constituents of Berberis aristata, Indian Journal of Chemistry6: Singh J, Kakkar P, Antihyperglycemic and antioxidant effect of Berberis aristata root extract and its role in regulating carbohydrate metabolism in diabetic rats,J Ethnopharmacol ,