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The Antimicrobial Activity and Brine Shrimp Lethality Bioassay of Leaf Extracts of Stephania Japonica (akanadi)

Rahman M Minhajur
Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali-3802, Bangladesh,

M Masud Alam
Department of Microbiology, Noakhali Science and Technology University, Sonapur, Noakhali-3802, Bangladesh,

A F M Shahid-Ud-Daula
Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali-3802, Bangladesh,

Mohammad Shahriar
Department of Microbiology, University of Asia Pacific,Dhanmondi-6A, Dhaka, Bangladesh

Md Mizanur Rahman Moghal
Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali-3802, Bangladesh,

Rokeya Siddiqui
Department of Microbiology, Noakhali Science and Technology University, Sonapur, Noakhali-3802, Bangladesh,

Abstract/ Executive Summary:

The purpose of the present study is to examine the antibacterial and cytotoxic properties of methanol extract of leaves of Stephania japonica. The crude methanolic extract of S. japonica, n-hexane, chloroform and ethyl acetate soluble fractions of methanolic extract were screened for their antimicrobial activity against a wide range of both Gram-positive and Gram-negative bacteria by disc diffusion method. The crude extract showed moderate and n-Hexane, chloroform soluble fraction of crude extract showed mild antibacterial activity against both Gram-positive and Gram-negative bacteria. Ethyl acetate soluble fraction of the extract showed significant antimicrobial activity against Salmonella typhi, Escherichia coli and Bacillus cereus. The zones of inhibition produced by the crude methanolic extract, n-hexane, chloroform and ethyl acetate soluble fractions were found to be 12.80-16.55 mm, 12.60 mm, 5-14.30 mm and 10-20.25 mm, respectively, at a concentration of 30 g/disc. Chloroform, n-hexane and ethyl acetate soluble fractions of methanolic extract of S. japonica were screened for antitumor properties using brine shrimp lethality bioassay. A reputed cytotoxic agent vincristine sulphate was used as a positive control. From the results of the brine shrimp lethality bioassay, it can be well predicted that chloroform and ethyl acetate soluble fractions of methanolic extract of S. japonica possess cytotoxic principles (with LC50 66.488 mg/ml and LC50 45.662 mg/ml, respectively) comparison with positive control vincristine sulphate (with LC50 0.839 mg/ml). But n-hexane soluble fractions of methanolic extract of S. japonica exhibited no lethality effect on shrimp nauplii

Keywords: Stephania japonica, Antimicrobial activity, Brine shrimp lethality bioassay

Location:

Start Date:

End Date:

Program Area: Pest Management

Commodity: Shrimp

Objectives:

To examine the antibacterial and cytotoxic properties of methanol extract of leaves of Stephania japonica.

Methodology:

Bacterial Strains: Six Gram positive and Gram negative bacterial strains were obtained from the stock culture of the Department of Pharmacy, University of Asia Pacific, Bangladesh . These strains were grown in nutrient broth (Difco Laboratories, USA) at 37°C and maintained on nutrient agar slants at 4°C

Plant materials: The plant S. japonica was collected from the Noakhali district, Bangladesh. The plant was identified by the Bangladesh National Herbarium, Dhaka, and a voucher specimen (Accession No: 35981) was deposited at the Department of Pharmacy, Noakhali Science and Technology University. The leaves were collected from the fresh tree. And then dried in an oven tray for 2 days at 40°C to 45°C. After drying, the plant part was ground into fine powder, which was almost red in color

Preparation of extracts: The plant extract was made by cold extraction method. Four hundred gram of powder was soaked in 1300 ml of 90% methanol. The container with its contents was sealed and kept for a period of 7 days with occasional shaking and stirring. The whole mixture was then filtered a coarse piece of clean, white cotton fabric. Then it was filtered through Whatman filter paper No-3. and the filtrate thus obtained was concentrated by evaporation and dried to a solid in an oven (Memmert: DIN12880-KI). Finally, reddish colour primary methanolic extract was found. Next, 5 g of primary methanolic extract was dissolved in 100 ml of 90% methanol. This is the mother solution, which was partitioned off successively by solvent-solvent extraction methods with three solvents of different polarity such as n-hexane, ethyl acetate, and chloroform .

Antibacterial Screening: The antimicrobial activity of the methanolic, n-hexane, ethyl acetate and chloroform extracts of S. japonica leaves sample were evaluated using agar disc-diffusion according to the Kirby Bauer method against the six Gram positive and Gram negative test organisms. The final concentrations of test samples were 3 μg/μl by dissolving the test samples with specific volume of solvents .Sterile Whatman filter paper discs (5 mm in diameter) were taken in a petridish. One drop of sample solution of the 3 μg/μl concentration was applied on the discs with the help of a micropipette in an aseptic condition. These discs were left for few minutes in aseptic condition for complete removal of solvent. Kanamycin (30 μg/disc) standard discs were used as a positive control. Blank discs and one drop of disc containing each solvent were used as a negative control.

Source of Information: Bangladesh J Microbiol, Volume 28, Number 2, December 2011, pp 52-56

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

From the results of the brine shrimp lethality bioassay, it can be well predicted that chloroform and ethyl acetate soluble fractions of methanolic extract of S. japonica possess cytotoxic principles (with LC50 66.488 mg/ml and LC50 45.662 mg/ml, respectively) comparison with positive control vincristine sulphate (with LC50 0.839 mg/ml). But n-hexane soluble fractions of methanolic extract of S. japonica exhibited no lethality effect on shrimp nauplii.

Knowledge Management: Journal