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Phytochemical and Biological Activities of Ethanolic Extract of C. Hirsuta Leaves

M. A. Rahman
Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong-4331, Bangladesh

M. A. Rahman
Bangladesh Council of Scientific and Industrial Research, Rajshahi

N. U. Ahmed
Bangladesh Council of Scientific and Industrial Research, Rajshahi

Abstract/ Executive Summary:

This study investigated the phytochemical and biological activities like antioxidant, antibacterial and cytotoxitc effects of C. hirsuta (Synonym- Senna hirsuta) leaf extracted with exhanol. Qualitative phytochemical analyses were accomplished by established methods. Antioxidant, antibacterial and cytotoxic effects were, respectively, measured by DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging method, disc diffusion method and brine shrimp lethality bioassay. The presence of alkaloids, glycosides, anthraquinone glycosides and flavonoids were detected in the leaf extract. The extract showed significant (p < 0.01) radical scavenging effect (IC50, 200.96 ± 0.85 μg/ml) in comparison to ascorbic acid (IC50, 1.24 ± 0.08 μg/ml). The extract also showed significant (p < 0.05) zone of inhibitions against Gram-positive Bacillus cereus (13 ± 1.05 mm) and Bacillus megaterium (9 ± 0.85 mm), and Gram-negative Vibrio cholera (20 ± 0.68 mm), Escherichia coli (8 ± 0.92 mm), Pseudomonas aeroginosa (15 ± 1.03 mm), Salmonella paratyphi (9 ± 0.79 mm) and Shigella dysenteriae (8 ± 0.88 mm). In cytotoxic assay, the extract showed the LC50 value 315.5 μg/ml which was statistically significant (p < 0.01) compared to reference cytotoxic agent vincristine sulfate (LC50, 38.99 μg/ml).

Keywords: Cassia hirsuta; DPPH radical scavenging; Antibacterial; Cytotoxic; LC50 Value; IC50Value.

Location: Chittagong University Campus

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

This study explored the free radical scavenging effect, antibacterial and cytotoxic effect of C. hirsuta leaf ethanolic extract.

Methodology:

Plant material: Plant materials were collected from Chittagong University Campus. The plant was taxonomically identified. A voucher specimen is preserved in Bangladesh National Herbarium with the accession No. BCUA- 03/12. Chemicals and reagents: Absolute alcohol and 1,1-diphenyl-2-picrylhydrazyl (DPPH) was purchased from Sigma-Aldrich, Munich, Germany. Tetracycline disc (30 μg/disc, Oxoid, England) and ascorbic acid (BDH, England) were used as reference standards for antibacterial screening and free radical scavenging assay, respectively. Vincristine sulfate (Merck, Germany) was used as reference cytotoxic agent in brine shrimp lethality test. Preparation of leaf extract: The fresh leaves of C. hirsuta were washed with deionized water immediately after collection. Air dried leaves were ground into powder (600 g) to be macerated in absolute ethanol for 7 days at room temperature (23 ± 5) °C. Extract was filtered through Whatman No. 1 filter paper (32cm) and concentrated under reduced pressure at the temperature below 50°C through rotatory vacuum evaporator (RE200, Sterling, UK). The crude extract (72 g blackish-green colored, 12% w/w yield) was stored at 4°C for further use. Phytochemical screening: The plant extract was phytochemically screened for the qualitative detection of alkaloids, glycosides, cardiac glycosides, anthraquinone glycosides, terpenoids and flavonoids using standard techniques as described below: Tests for alkaloids: The alkaloids test has been done according to the method described by El-olemy et. al. (1994). To conduct this test, a 0.5 g of extract was stirred with 5 ml 1% HCl on a steam bath and then the solution was cooled and filtered. 1ml of the filtrate was treated separately with drops of Mayer's, Hager's and Wagner's reagents and formation of dirty/dark brown, yellow-brown or reddish brown precipitate, respectively, indicated the presence of alkaloids. Test for Glycoside: The presence of glycosides in the plant extract was confirmed by general tests as described by Sofowora (1984). General test: A 500 mg of an ethanolic extract was dissolved in one ml of water. A few drops of aqueous NaOH solution were added to the solution. Development of yellow color indicated the presence of glycoside. Test for cardiac glycosides: The presence of cardiac glycosides and O-glycosides in the plant extract was confirmed by Legal's test and Baljet testsas described by Sofowora (1984). I Legal's test: A 100 mg of an ethanolic extract of the plant material was dissolved in two ml of pyridine and two ml of sodium nitroprusside solution (0.5%). The mixture was alkalined with NaOH (0.2 N) solution. Development of pink to red color development indicated the presence of cardiac glycoside due to lactone ring. II Baljet test: A drop of Baljet's reagent (95 ml 1% picric acid + 5 ml 10% NaOH) was added to a two ml of an ethanolic extract of plant material. Development of yellow orange color development indicates the presence of five membered lactone rings at C-17 of the aglycone in cardiac glycoside.

Source of Information: Bangladesh J. Sci. Ind. Res. 48(1), 43-50, 2013

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

This study demonstrates the potentials of the C. hirsuta extract in free-radical scavenging, bacterial inhibition and cytotoxicity. Antibacterial and cytotoxic effect of the extract lead to the possession of different active phytochemicals in this extract and their individual or synergistic function could also be delineated from this study. Future study is required to isolate and characterize the individual bioactive compounds as well as to elucidate the possible mechanisms lying with these effects.

Knowledge Management: Journal