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Antimicrobial and Cytotoxic Activities of Root Extracts of Piper Chaba

Tarannum Naz
Department of Pharmacy, University of Rajshahi, Rajshahi 6205, Bangladesh

Ashik Mosaddik
Department of Pharmacy, University of Rajshahi, Rajshahi 6205, Bangladesh

M. Ekramul Haque
Department of Pharmacy, University of Rajshahi, Rajshahi 6205, Bangladesh

Abstract/ Executive Summary:

Antibacterial and antifungal properties of petroleum ether, chloroform, ethyl acetate and methanol extracts of Piper chaba (Choi) roots were studied by disc diffusion method and these activities were compared with primary standard drugs Kanamycin and Nystatin, respectively. The extracts were found to exhibit promising antibacterial and antifungal properties against Gram-positive, Gram-negative bacteria and fungi. The extracts were also studied for their cytotoxic activities by brine shrimp lethality bioassay, where gallic acid was used as primary standard. It was observed that the petroleum ether extract was potent cytotoxic with the LC50 value of 0.95 μg/ml against Artemia salina (L). The essential oils of the petroleum ether extract of Piper chaba roots were analysed by GC/MS. It was observed that most of the compounds were sesquiterpenes, some were long chain fatty acids and some were monoterpenes and alkaloids. Seventeen compounds were identified from the GC/MS analysis.

Keywords: Piper chaba, Antibacterial activity, Antifungal activity, Cytotoxicity.

Location: Department of Pharmacy, University of Rajshahi, Rajshahi 6205, Bangladesh

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

In this paper it was reported antimicrobial and cytotoxic activities of the crude extracts of the roots of Piper chaba and the compositions of the essential oils of petroleum ether extract.

Methodology:

Plant collection and extraction: Roots of Piper chaba were collected from the Bagerhut District, Bangladesh. The plant was botanically identified by Prof. A. T. M. Naderuzzaman, Department of Botany, University of Rajshahi. A voucher specimen was deposited in Bangladesh National Herbarium (Voucher No. 30276). The extraction was carried out by soaking the dried, grounded roots (1 kg) of Piper chaba in 5 liter rectified spirit for two weeks at room temperature. Evaporation of the solvent under reduced pressure afforded a semi-solid mass (40 gm). The crude extract was successively partitioned with petroleum ether, chloroform, ethyl acetate and finally with methanol. Solvents of these extracts were concentrated separately by evaporation under reduced pressure below 50ºC, which yielded 10.0 g, 12.3 g, 1.2 g and 5.6 g of the dried extracts, respectively. Test organisms: Four Gram-positive bacteria (Bacillus subtilis, Bacillus megaterium, Staphylococcus aureus, Strep. β-haemolyticus) and six Gram-negative bacteria (Escherichia coli, Salmonella typhi, Shigella dysenteriae, Shigella shiga, Klebsiella spp., Shigella boydii) were used for antibacterial activity study, four fungi (Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, Candida albicans) were used for antifungal activity study and a simple zoological organism (brine shrimp nauplii) was used for cytotoxicity study. All organisms were collected from the stock culture of the Institute of Nutrition and Food Sciences, University of Dhaka, Bangladesh. Antibacterial, antifungal and cytotoxicity studies: Antibacterial and antifungal activity studies were carried out by disc diffusion method and cytotoxicity was studied by brine shrimp lethality bioassay. Standard antibiotic disc of Kanamycin (K-30 μg/disc) was used as the standard reference drug for antibacterial assay, Nystatin (50 μg/disc) was used for antifungal activity study and gallic acid was used as a standard reference for cytotoxicity study. Chemical analysis by GC/MS: The essential oils were examined by ISO-ext method in Agilent, 6890 series-GC system (made in USA). ISO-ext method is a long routine method to determine non-polar to polar compounds in a mixture. In this analysis, total running time was 73 min from 50°C (1 min hold) to maximum 300°C (10 min hold) and ramp time was 4 min. Petroleum ether extract (1 gm) of Piper chaba root was dissolved in small amount of acetone and shaken in UV-water bath. Then acetone soluble fraction was collected and dried off by Nitrogen gas to afford an oily mass for the GC/MS analysis. 5 mg of acetone soluble fraction was dissolved in 1 ml of acetone. For this analysis, 1 μl of samples was injected in split mode and selective mass detector with helium gas at 1.6 ml/min under 31.7 psi pressures and AV with 35 cm/sec were used.

Source of Information: J. Sci. Res. 1(1), 138-144 (2009)

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Funding Source:

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Achievement/Findings:

The ethyl acetate extract of Piper chaba root shows a significant antibacterial activity, followed by methanol extract against both Gram-positive and Gram-negative bacteria. This result suggests that the root extracts of Piper chaba can be used for the treatment of fever, bronchitis and tuberculosis. The petroleum ether and chloroform extracts show moderate antibacterial activity but they exhibit very potent cytotoxic property with LC50 values of 0.95 and 1.53 μg/ml, respectively, compared to standard gallic acid. This result supports the use of Piper chaba as anthelmintic in traditional medicine. All extracts show moderate antifungal activity. From GC/MS analysis, it is clear that the oily mass of petroleum extract of Piper chaba (root) possesses some sesquiterpenes plus some intermediate polar compounds which might be interesting for further isolation and investigation of biological activity.

Knowledge Management: Journal