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Phytochemical Screening, Antimicrobial and Anticancerous Activities of Two Different Plant Extracts

Humaira Gulzar
A) Department of Pharmacy, East West University, Dhaka-1212, Bangladesh. B) Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh.

Md Irfan Amin Chowdury*
Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh

Mohammad Nazmul Alam
Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh.

SM Arif Bin Alam
Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh.

Mohammad Abu Sayem Shakil
Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh.

Md Mahidur Rahman Khan
Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh

Md Mizanur Rahman
Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh.

Muhammad Moin Uddin Mazumdar
Department of Pharmaceutical Sciences, North South University, Dhaka-1229, Bangladesh.

Abstract/ Executive Summary:

The phytochemical screening of ethanolic leaf extract of L. glutinosa and C. quadrangularis confirmed the presence of alkaloids, steroids, flavonoids, terpenoids, tannins, anthraquinones and alkaloids, flavonoids, terpenoids respectively. Disk diffusion method was performed to evaluate the antimicrobial activity of these plant extracts. L. glutinosa showed its highest activity against a gram negative strain S. paratyphi which exhibited a ring of 12 mm in diameter at 400 μg/disc. In the case of C. quadrangularis, highest activity was found against S. paratyphi which showed a ring of 12.33 mm in diameter at 400 μg/disc. On the contrary, standard Kanamycin showed its highest zone of inhibition against E. coli which is 28.66 mm in diameter at 30 μg/disc. In the case of anticancerous activity, standard vincristine sulphate showed 12.59 µg/ml activity whereas ethanolic leaf extract of L. glutinosa & C. quadrangularis demonstrated minimal activity which is 530.05 µg/ml and 639.68 µg/ml respectively.

Keywords: Litsea glutinosa, Cissus quadrangularis, Phytochemical screening, Disk diffusion method, Kanamycin, Vincristine sulphate

Location: Bangladesh

Start Date:

End Date:

Program Area: Resource Development and Management

Commodity: Extract (plant, seed)

Objectives:

Stem juice is used in otorrhoea, epistaxis, scurvy and irregular menstruation. Paste of stem is given in asthma; stem boiling in limewater is a useful stomachic. Stems, roots and leaves are used as a plaster over broken limbs. Burnt to ashes of the young shoots administered in dyspepsia and other bowel complaints. Leaves and young shoots are considered as powerful alteratives. EtOH (50%) extract of aerial parts is hypotensive and diuretic.

Methodology:

Plant material Litsea glutinosa & Cissus quadrangularis were collected from local area of Chittagong district (Sitakunda), Bangladesh and authenticated by the Botanist Dr. Shaikh Bokhtear Uddin, Associate Professor, Department of Botany, University of Chittagong, Bangladesh. Preparation of extraction The leaves were indirectly sun dried by shade and ground. The ground (500g) were soaked in sufficient amount of ethanol (1:4) for one week at room temperature with occasional shaking and stirring then filtered through a cotton plug followed by Whitman filter paper No. 1. The solvent were evaporated under vacuum at room temperature to yield semisolid. Ethanolic leaf extract was then preserved in a refrigerator at 4 ºC till further use. Chemicals and reagents Methanol and Kanamycin (30 μg/disc) were purchased from Merck (Germany) and Oxoid (England) respectively. Two more chemicals were purchased from Sigma Aldrich (Munich, Germany), which were vincristine sulfate and 99.5% absolute ethanol. All other reagents were of analytical grade. Phytochemical screening Chemical tests were carried out on the ethanolic extracts of Litsea glutinosa and Cissus quadrangularis using standard procedures to identify the constituents as described by Sofowora (1993), Trease and Evans (1989) and Harborne (1973). Preparation of Test Sample Solutions 20mg of each plant extract was added to 10ml Methanol. Sonicator was used to dissolve the extracts. The solution was then filtered. The method was described by Savithramma, Rao, & Surhulatha, 2011. Test for Alkaloids Preparation of Dragendroff’s Reagent Step-01 Solution A - 0.17gm Bismuth nitrate was added in 2ml Acetic acid and 8ml Distilled water. Step-02 Solution B - 4gm Potassium iodide was added in 10ml Acetic acid and 8ml Distilled water. Step-03 Dragendroff’s Reagent - Solution A and B were added and mixed together. The mixture was then diluted with distilled water up to 100ml. Procedure 2ml of filtrate in another was taken in a test tube and 1% Hydrochloric acid was added to it and mixed properly. 1ml from the mixture was taken and 6 drops of Dragendroff’s reagent was added to it. This method is identified and examined by Harborne, 1998. Test for Flavonoids 5 ml of dilute ammonia solution were added to 4ml of filtrate of each plant extract followed by addition of 1ml concentrated H2S04. Test for Steroids 1ml of chloroform and 1 ml of concentrated H2S04 was added to 4ml filtrate of each plant extract. Test for Terpenoids 0.5ml Acetic anhydride and 0.5ml chloroform was added to 4mg of each plant extract [8]. Test for Reducing-sugars 1ml of distilled water was added to 0.5ml of filtrate of each plant extract. Then 5-8 drops of Fehling’s solution A & B were added in equal amounts. The mixture was then heated. Test for Tannins About 0.5 g of the dried extract of Litsea glutinosa was boiled in 20 ml of distilled water and then filtered. A few drops of 0.1% ferric chloride were added. Test for Anthraquinone 0.5gm of dried extract of Litsea glutinosa was boiled with 10ml concentrated H2S04. The solution was then filtered while hot. The chloroform layer was pipette into another test tube and 1ml ammonia solution was added to the chloroform layer. Test for Cardiac glycosides (Keller-Killani test) Five ml of each extracts were treated with 2 ml of glacial acetic acid containing one drop of ferric chloride solution. 1 ml of concentrated sulphuric acid was then added to it. Antimicrobial activity Test Materials In this present study, the antibacterial activity of ethanolic extracts of Litsea Glutinosa and Cissus quadrangularis was investigated in comparison with standard kanamycin (30 μg/disc) antibiotic agent against a number of pathogenic Gram-positive and Gram-negative bacteria and fungus. Test Organisms Both Gram-positive and Gram-negative strains of bacteria and fungus were used as the test organism to observe the antibacterial activity of the isolated compounds. These organisms were collected from the Microbiology research laboratory, Department of Pharmacy, East West University. The pure of which was previously collected from the Department of Microbiology, University of Dhaka. Preparation of the media The instant nutrient broth was prepared from purified powdered agar, which were weighed and then reconstituted with distilled water in a conical flask according to specification. It was then heated in a water bath to dissolve the agar until a transparent solution was obtained. In this way nutrient broth was prepared.

Source of Information: Journal of Medicinal Plants Studies 2015; 3(6): 76-81

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Achievement/Findings:

The result of phytochemical screening revealed that, ethanolic leaf extract of L. glutinosa posses alkaloids, steroids, flavonoids, terpenoids, tannins, anthraquinones & C. quadrangularis contain alkaloids, flavonoids and terpenoids. Ethanolic leaf extracts of L. glutinosa and C. quadrangularis possess moderate antimicrobial activity. Those plants are also exhibited minimal anticancerous activity.

Knowledge Management: Journal