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Research Article
 

In vitro Antibacterial Activity of the Extracts and a Glycoside from Sida rhombifolia Linn



Md. Ekramul Islam, Naznin Ara Khatune and Md. Ekramul Haque
 
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ABSTRACT

The research work was conducted to investigate the in vitro anti-bacterial activity of a glycoside, phenyl ethyl β-D-glucopyranoside, isolated from the stem of the plant Sida rhombifolia. The petroleum ether (C2H5-O-C2H5), chloroform (CHCl3) and ethylacetate (CH3-CO-O-C2H5) extracts of the plant were screened against eleven pathogenic bacteria for their antimicrobial activities. The test materials were found to be significant in vitro antibacterial activities, against most of the test bacteria. The zones of inhibition produced by the test materials were found to be between 8 and 24 mm. The minimum inhibitory concentration (MIC) values of the isolated compound was also determined against Bacillus subtilis, Sarcinia lutea, Escherichia coli and Shigella shiga which were 128, 64, 64 and 128 ยตg ml-1, respectively.

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  How to cite this article:

Md. Ekramul Islam, Naznin Ara Khatune and Md. Ekramul Haque , 2002. In vitro Antibacterial Activity of the Extracts and a Glycoside from Sida rhombifolia Linn. Journal of Medical Sciences, 2: 134-136.

DOI: 10.3923/jms.2002.134.136

URL: https://scialert.net/abstract/?doi=jms.2002.134.136

Introduction

Malvaceae is a cosmopolitan family of herbs, shrub and trees (Heywood, 1978). Modern research carried out on the malvaceous plants revealed that most of the plants belonging to this family are medicinally important as they contain biologically active compounds. Sida rhombifolia locally known as Berela in Bangladesh belongs to the family Malvaceae. It has considerable reputation for its medicinal value in traditional medicine. The roots and stems are useful in fever, heart disease, piles and all kinds of inflammation. An infusion of the root is given in dysentery. Stem is also employed as demulcent and emollient (Kirtikar and Basu, 1993). The plant is also useful in tuberculosis and used in skin diseases and as a diuretic (Colonel, 1986). From the literature survey, it is revealed that the preliminary screening of methanolic extract (CH3OH) of the leaves of the plant Sida rhombifolia, showed anti-tumour and anti-HIV activity (Muanza et al., 1995). The ethanolic (C2H5OH) extracts of this plant showed inhibitory activity on NF-kB cell line as an anti–inflammatory model (Bork et al., 1996).

Although Sida rhombifolia is locally used for the above conditions, no antibacterial study of this plant has previously been reported. As a part of continuing search for novel antibacterial principles from the medicinal plants of Bangladesh, studied, Sida rhombifolia and herein report the results of in vitro antibacterial investigation.

Materials and Methods

Plant materials: Matured stems of the plant were collected from Rajshahi University Campus, Rajshahi, Bangladesh, during the month of November-December 1999. The plant was taxonomically identified by the Department of Botany, Rajshahi University as well as Bangladesh National Herbarium, where a voucher specimen is kept.

Extraction, isolation and characterization: The sun dried stems were put in an oven at 45 0C, crushed into power with a crushing machine and extracted in soxhlet apparatus with methanol (CH3OH) for 72 hours at it’s boiling point (64.5 0C ) (Morrison and Boyd, 1996). The concentrated methanolic extract was diluted with distilled water and solvent-solvent partitioning was successfully carried out by Kupchan method (Grode et al., 1983) using petroleum ether (C2H5-O-C2H5), chloroform (CHCl3) and ethylacetate (CH3-CO-O-C2H5) (Islam, 2000). Each of the extract was concentrated at reduced pressure using rotary evaporator and thus ready for antibacterial screening.

The compound, phenyl ethyl β-D-glucopyranoside was isolated from the chloroform fraction by column chromatography (Beckett and Stenlake, 1986) followed by preparative thin layer chromatography (PTLC) (Egon and Stahl, 1969).

Image for - In vitro Antibacterial Activity of the Extracts and a Glycoside from Sida rhombifolia Linn

This compound was characterized on the basis of its 1 H-NMR, 3C-NMR, 1H-1H COSY 900 (Islam, 2001). Then this compound was subjected to antibacterial screening.

Antibacterial Screening: Eleven pathogenic bacteria (four gram positive and seven gram negative) were selected for the test and collected from the Department of Microbiology, Dhaka University, Dhaka, Bangladesh. Nutrient agar was used as a bacteriological medium. The petroleum ether (C2H5-O-C2H5), chloroform (CHCl3) and ethylacetate (CH3-CO-O-C2H5) extracts were dissolved separately in sufficient amount of methanol to get a concentration of 500μg 10 μl-1. The compound, Phenyl ethyl β-D-glucopyranoside was also dissolved separately in methanol in the same way to get a concentration of 300μg 10μl-1. Then in vitro antibacterial activity of these samples were carried out by the standard disc diffusion method (Barry, 1980; Berghe and Vlietnck, 1991; Bauer et al., 1996; Rios et al., 1988) against selected test organisms. The diameter of zone of inhibition produced by the extracts and pure compound, Phenyl ethyl β-D-glucopyranoside was then compared with those produced by the standard antibiotic (Kanamycin 30 μg disc-1).

Minimum inhibitory concentration (MIC): The minimum inhibitory concentration (MIC) of the compound, Phenyl ethyl β-D-glucopyranoside was determined against two gram positive (Bacillus subtilis and Sarcina lutea) and two gram negative (Escherichia coli and Shigella shiga) bacteria. The tests were carried out by serial dilution technique (Reiner, 1982). Nutrient agar and nutrient broth were used as bacteriological media.

Results and Discussion

The antibacterial activities of the petroleum ether (C2H5-O-C2H5), chloroform (CHCl3) and ethylacetate (CH3-CO-O-C2H5) extracts and the pure compound, phenyl ethyl β-D-glucopyranoside, isolated from the chloroform extract of Sida rhombifolia stem against eleven pathogenic bacteria (Table 1). The zone of inhibition of petroleum ether, chloroform and ethylacetate extract were found to be 11-18, 11-22 and 12-24 mm at a concentration of 500 μg disc-1, respectively. The compound, Phenyl ethyl β-D-glucopyranoside produced zone of inhibition between 9 and 14 mm, at a concentration of 300 μg disc-1.

Table 1: Antibacterial activities of extracts, pure compound, Phenyl ethyl β-D-glucopyranoside and Kanamycin standard
Image for - In vitro Antibacterial Activity of the Extracts and a Glycoside from Sida rhombifolia Linn
Image for - In vitro Antibacterial Activity of the Extracts and a Glycoside from Sida rhombifolia Linn

The crude extracts and the glycosidal compound, phenyl ethyl β-D-glucopyranoside showed significant activity against the bacteria tested. Chloroform extract showed more activity than ethylacetate extract against most of the test bacteria and in particular against Bacillus subtilis and Staphylococcus aureus (Gram positive) and Shigella dysenteriae and Shigella sonnei. Ethylacetate extract showed highest activity against Sarcina lutea, Shigella shiga and Pseudomonas aeruginosa but petroleum ether extract has shown the lowest activity in most of the test bacteria.

Table 2: The MIC values of the isolated compound against test organisms
Image for - In vitro Antibacterial Activity of the Extracts and a Glycoside from Sida rhombifolia Linn

Minimum inhibitory concentration (MIC) values of the compound, phenyl ethyl β-D-glucopyranoside were 64 μg ml-1 against Sarcina lutea and E. coli and 128 μg ml-1 against Bacillus subtilis and Shigella shiga (Table 2).

From minimum inhibitory concentration (MIC) values, it was found that this glycosidal compound was more potent against Sarcina lutea and Escherichia coli possessing less minimum inhibitory concentration (MIC) values against Bacillus subtilis and Shigella shiga.

In conclusion, the study reports for the first time the antibacterial activity for the crude extracts and the isolated compound from the stem of Sida rhombifolia. However, further and specific studies are needed to better evaluate the potential effectiveness of other isolated compound from this plant.

Acknowledgments

The authors would like to thank the Department of Microbiology, University of Dhaka, Bangladesh and the Microbiological Laboratory of the Institute of Nutrition and Food Science (INFS) for supply of test organisms. We are also indebted to Professor A.T.M. Naderuzzaman, Department of Botany, Rajshahi University, Bangladesh and Bangladesh National Herbarium for the identification of the plant.

REFERENCES

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9:  Islam, M.E., 2001. Chemical and biological studies on Sida rhombifolia Linn. M.Sc. Thesis, Department of M.SC. Pharmacy, University of Rajshahi, Bangladesh.

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