Evaluating Cassia tora Linn. Seeds for their Antimicrobial Potential with Special Reference to Skin Infections

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About Author: Saurabh Jain.*1, M.D.Kharya and Jain Aakansha2
1 Department of Pharmaceutical Sciences
2 Department of Microbiology
Dr. H. S. Gour Central University Sagar (M.P.), India.470003

Abstract
Due to increasing resistance to existing synthetic antimicrobial agents, herbal drugs are being looked as importance source for treating various ailments related to skin infections and also for the discovery of new antimicrobial molecules for their synthesis. Cassia tora is a well known plant possessing wide range of pharmacological activities. It has been used in India as folk remedy in the form of decoctions and infusions to treat bacterial and fungal infections and also claimed to be effective against variety of skin conditions like psoriasis, acne, wounds etc. Although this plant, particularly its seeds have been reported to have antimicrobial activities, but no targeted and specific study has been reported. The present investigation was carried out to study this unexplored area of this drug. As the seeds of C. tora possess good antimicrobial activity, it was thought worked to scientifically explore its potential on microorganisms effecting human beings especially on skin, so that safe and effective herbal treatment can be developed for the same.

Reference ID: PHARMATUTOR-ART-1108

Introduction
Traditional medicines hold a great promise as source of easily available effective therapy for skin diseases. Cassia tora Linn.(Family: Leguminosae), is a common Indian herb having various medicinal properties for the treatment of many disease. According to Ayurveda the leaves and seeds of C.tora are acrid, laxative, antiperiodic, antihelmintic, ophthalmic, liver and cardiotonic and expectorant and useful in leprosy, ringworm, flatulence, colic, dyspepsia, constipation, cough, bronchitis and cardiac disorders. It has been reported for their usefulness in traditional system of medicines for treating skin diseases like psoriasis, leprosy etc (Zahra et al, 2000; Harrison and Dorothy, 2003). C.tora is used as natural pesticides. Antimicrobial activity of chrysophanic acid-9-anthrone from C. tora has been reported. (Kim et al, 2004). Humans are natural hosts for many bacterial species that colonize the skin as normal flora. Staphylococcus aureus and Streptococcus pyogenes are infrequent resident flora, accounting for a wide variety of bacterial pyrodermas, conditions like impetigo, ecthyma, erysipelas, cellulitis, psoriasis, atopic dermatitis, Erythroderma and other skin infections (Tomi et al, 2005). As C.tora has been reported to possess antimicrobial activity causing skin infections, the present investigation was carried out to evaluate the antimicrobial activity of Cassia tora, against Staphylococcus aureus, Escherichia coli, Aspergilus niger and Microsporum gypseum.

Materials and Methods:
Seeds of C. tora collected from local market of Sagar, were dried and authenticated in Department of Botany, Dr. H.S.Gour Central University, Sagar (M.P.), India.

Chemicals and Reagents: All the solvents used were of AR grade from Rankem.

Apparatus: Kitchen blender, Soxhlet apparatus, sieve size #10.

Extraction of Cassia tora: Seeds were washed in tap water; shades dried, powdered in a kitchen blender and were stored in airtight plastic bags. Then powdered seeds (250g.) passed through sieve # 10 was defatted by the Whatman filter paper and introduced in to the Soxhlet apparatus using petroleum ether (60-80) as a solvent. After complete defatting drug was air dried for removing trace of petroleum ether. The defatted drug was then packed in Whatman filter paper and introduced in to the Soxhlet apparatus and extracted with benzene as a solvent for complete extraction. The extract was filtered, concentrated and dried in a water bath. Dried extract was transferred to air tight bottles and the percentage yields was calculated and stored at cool place. (Table-1).

Isolation of Chrysophanic acid: Isolation of chrysophanic acid was done by column chromatography of benzene extract. The column was washed three times with acetone and packed with activated silica gel (100-120 mesh). The elution was started with the chloroform solvent system. Since most of the components were yellowish- brown coloured, it was easy to govern the elution pattern. Each fraction collected separately and concentrated by evaporation. There were around twelve fractions and each fraction was compared with its adjacent fractions by TLC.  Those fractions given similar pattern combined together and each fraction qualitatively analyzed for the particular class of compound. Then after, obtained isolated compound was identified by various methods as apperance, melting point, solibilty profile, Modified Borntrager’s test and UV-visible spectra range etc.

Phytochemical evaluation: Phytochemical examinations were carried out for the extract as per the standard methods. (Trease and Evans, 1987). (Table-2)

S.No.

Phytochemicals

Test Procedure

1.

Alkaloids

Extract was dissolved in dilute Hydrochloric acid and filtered. The filtrate was used to test for the presence of alkaloids.

Mayer’s Test

Filtrate was treated with Mayer’s reagent (Potassium Mercuric iodide). Formation of a yellow cream precipitate indicates the presence of Alkaloids.

Dragendroff’s Test

Filtrate was treated with Dragendroff’s reagent (solution of potassium bismuth iodide). Formation of red precipitate indicates the presence of alkaloids.

2.

Carbohydrates

Extract was dissolved in 5 ml distilled water and filtered. The filtrate was used to test for the presence of carbohydrates.

Molisch’s Test

Filtrate was treated with 2 drops of alcoholic α-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.

3.

Glycosides

Extract was hydrolysed with dil. HCl, and then subjected to test for glycosides.

Modified Borntrager’s Test

Extract was treated with 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 ammoniacal layer indicates the presence of anthraquinone glycosides.

Legal’s Test

Extract was treated with sodium nitroprusside in pyridine and methanolic alkali. Formation of pink to blood red colour indicates the presence of cardiac glycosides.

4.

Saponins

Froth Test

Extract was diluted with distilled water to 20ml and this was shaken in a graduated cylinder for 15 minutes. Formation of 1 cm layer of foam indicates the presence of saponins.

5.

Steroids

Libermann Burchard’s Test

Extract was treated with chloroform and filtered. The filtrate was treated with few drops of acetic anhydride, boiled and cooled. Conc. Sulphuric acid was added carefully along the sides of the test tube. Formation of brown ring at the junction indicates the presence of steroids

6.

 

 

Fixed oils & fats

Stain Test

Small quantity of extract was pressed between two filter papers. An oily stain on filter paper indicates the presence of fixed oil.

 

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