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In vitro Antimicrobial Activity of the Compound Isolated from Chloroform Extract of Moringa oleifera Lam.



Farjana Nikkon, Zahangir Alam Saud , M. Habibur Rahman and Md. Ekramul Haque
 
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ABSTRACT

The research work was conducted to investigate the in vitro antimicrobial activity of the compound Aglycon of Deoxy-Niazimicine (N-benzyl,S-ethyl thioformate) isolated from Moringa oleifera Lam. in comparison with crude chloroform extract. Antibacterial and antifungal activities of the compound-1 and crude extract were observed against fourteen pathogenic bacteria and six pathogenic fungi. Compound-1 isolated from the chloroform (CHCl3) fraction of ethanol extract showed comparatively more in vitro antibacterial activity against Shigella boydii, Shigella dysenteriae and Staphylococcus aureus than the crude chloroform (CHCl3) extract and produced zone of inhibition in between 09 and 13 mm. The MIC values of the compound-1 and crude extract were also determined against B. megaterium, St. aureus, Sh. dysenteriae and Sh. boydii. The values were found to be between 32 to 128 μg ml-1. The compound-1 also showed moderate antifungal activity against Candida albicans and Aspergillus flavus than the crude chloroform extract although the crude extract showed moderate activity against Trichoderma species. Thus compound-1 has antibacterial and antifungal activity and suggested its therapeutic use as an antimicrobial agent.

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  How to cite this article:

Farjana Nikkon, Zahangir Alam Saud , M. Habibur Rahman and Md. Ekramul Haque , 2003. In vitro Antimicrobial Activity of the Compound Isolated from Chloroform Extract of Moringa oleifera Lam.. Pakistan Journal of Biological Sciences, 6: 1888-1890.

DOI: 10.3923/pjbs.2003.1888.1890

URL: https://scialert.net/abstract/?doi=pjbs.2003.1888.1890

INTRODUCTION

The plant Moringa oleifera Lam. locally known as shajna (English-drumstick) belongs to the family Moringaceae and is widely distributed in the Indo-Bangla subcontinent and cultivated through out the tropical belt (Sastri, 1962). Different parts of this plant are used in the indigenous systems of medicine for the treatment of a variety of human ailments and are also eaten as vegetable (Sastri, 1962; Chopra et al., 1956; Nadkarni and Nadkharni, 1976).

The targeted plant M. oleifera Lam. has many uses in traditional medicines. For example, root-barks are used as astringent, anthelmintic, analgesic, useful in heart complains, eye diseases, dyspepsia, enlargement of the spleen, tuberculous glands in the neck, tumors, earache, stuttering and in ulcers. The plant is also considered to be useful by tribals (Santals) in burns, sores, epilepsy, scabies, cholera, dysentery and retention of urine (Kirtikar and Basu, 1984).

The pharmacological activities of M. oleifera have extensively been studied, particularly the hypotensive action of the ethanolic extract of leaves (Dashputra et al., 1977), the antimicrobial activity of the seeds (Eilert et al., 1981; Caceres et al., 1991), the antifertility activity of roots (Prakash,1988; Shukla et al., 1988) and the spasmolytic, anti-inflammatory and diuretic properties of the leaves, fruits, barks and roots (Caceres et al., 1992). The present investigation reports on the isolation of Aglycone of Deoxy-Niazimicine (N-benzyl, S-ethyl thiocarbamate) from the chloroform fraction of ethanol extract of the root barks of M. oleifera Lam. and discuss about the antibacterial and antifungal activity of the compounds.

MATERIALS AND METHODS

The plant material: Fresh root barks of Moringa oleifera Lam were collected from the adjoining areas of Rajshahi University campus, Bangladesh in March, 2001 and identified by Professor A.T.M. Nadiruzzaman, Department of Botany, University of Rajshahi and a vaucher specimen (no. A Islam 3, collection date 02.05.1982) is deposited in the Department of Botany, Rajshahi University.

Extraction and isolation of compounds: The root barks were cut into small pieces, dried in the sun for 7 days and finally in an oven below 60EC. The dried plant material (1.5 kg) was ground into fine powder and exhaustively extracted with ethanol. The extract was concentrated to a dark brownish syrupy residue (50g) and then successively extracted with pet-ether (40-60°C) and chloroform. The organic layers were washed with water, dried over Na2SO4 and concentrated in vaccuo to afford a pet-ether soluble fraction (2g) and chloroform soluble fraction (6.9g).

Fig. 1:Aglycon of Deoxy-Niazimicine (N-benzyl, S-ethyl thioformate)

The chloroform soluble fraction (6.9g) was subjected to a column of silica gel-60 and eluted with ethyl acetate and n-hexane with increasing portions of ethyl acetate and finally with ethyl acetate, yielding several fractions. The fractions obtained with n-hexane-ethyl acetate (50:50) was rechromatographed eluting with n-hexane-CHCl3 with increasing portions of chloroform and finally with methanol. Fractions obtained with n-hexane-CHCl3 75:25 showed similar TLC profiles and were pooled. This fraction showed positive test for secondary amine and carbonyl function. Crystallization from pet-ether (40-60°C) affords a new compound-1 (31.4 mg) as color less needles (m.p. 58-60°C). Structure of this compound-1 was confirmed by analysis of its IR, 1H and 13C-NMR spectral data. (Fig. 1). The isolated compound-1 and the crude chloroform extract were subjected to antibacterial and antifungal screening.

Antimicrobial screening: In vitro antimicrobial screening is generally performed by disc diffusion method (Vander and Vlietnck, 1999) for primary selection of the compounds as therapeutic agent. Disc diffusion method is highly effective for rapidly growing microorganisms and the activities of the test drugs are expressed by measuring the diameter of the zone of inhibition. The standard test microorganisms were available in the microbiological research laboratory of Biochemistry and Molecular Biology Department, University of Rajshahi, Bangladesh. The pure cultures of these were collected from the Microbiological Laboratory of the Institute of Nutrition and Food Science (INFS) and Department of Microbiology, University of Dhaka, Bangladesh. For antibacterial activity Fourteen pathogenic bacteria (Five Gram positive and Nine Gram negative) were selected. The compound-1 and crude extracts were dissolved separately in CH3OH to get a concentration of 30 μg disc–1 and 100Fg disc–1, respectively. The diameter of zone of inhibition produced by both crude extract and compound-1 were then compared by the standard antibiotic Kanamycin 30 μg disc–1.

Minimum inhibitory concentration (MIC): The MIC values of the compounds were determined against three Gram positive and three Gram negative bacteria. The test was carried out by a serial tube dilution technique (Reiner, 1982). Nutrient agar was used as bacteriological media.

Antifungal activity :The antifungal activities of the compound-1 and crude extract were determined at a concentration of 50 μg disc–1 and 100μg disc–1 against six pathogenic fungi. Nystatin (50 μg disc–1) was used as standard.

RESULTS AND DISCUSSION

Compound -1 isolated from the chloroform fraction of ethanol extract from the root barks of the plants Moringa oleifera Lam., as colorless needles, melting point 58-60°C from pet-ether.

The antibacterial activity of the isolated compound-1 and crude extract against all tested organisms is shown in Table 1. Compound-1 showed strong antibacterial activity against gram positive Staphylococcus aureus and gram negative Shigella dysenteriae, Shigella boydii,Salmonella typhi and Pseudomonas aeruginosa and produced zone of inhibition between 09 to 13 mm.

Table 1:
In vitro antibacterial activity of compound-1 and crude extract Zone of inhibition (Diameter in mm)

Table 2:
The MIC values of the compound-1 and crude extract against test organisms Minimum inhibitory concentration in Fg ml–1

Table 3:
In vitro antifungal activity of the compound-1, crude extract and standard Nystatin. Zone of inhibition (diameter in mm.)

While the crude extract showed comparatively lower activity against gram positive bacteria Staphylococcus aureus and gram negative bacteria Shigella dysenteriae, Shigella boydii and produced zone of inhibition between 09 to 11mm. The MIC values of the isolated compound-1 and crude extract were determined against B. megaterium, St. aureus, Sh. dysenteriae and Sh.boydii shown in Table 2. Compound-1 showed moderate activity against Shigella dysenteriae, Shigella boydii, Staphylococcus aureus and MIC values were 32 μg ml–1. On the other hand, crude extract showed little activity against Shigella dysenteriae and the MIC value was 64 μg ml–1.

The antifungal activity of the compound-1 and crude extract were tested against six pathogenic fungi and the obtained result are cited in Table 3. Compound-1 showed moderate activity against Candida albicans and Aspergillus flavus in comparison with the standard Nystatin. But the crude extract showed only little activity against Trichoderma species.

Rao and George (Rao and George, 1949) reported that the oil extracted from the root of Moringa oleifera showed pronounced antibacterial activity. Kurup and Rao (Kurup and Rao, 1952) also found the antibacterial activity of the alcoholic extract of the root of Moringa pterygosperma against Micrococcus pyogenes (aureus). Moreover, Sen Gupta et al. (1956) reported the root bark of the plant Moringa pterygosperma has highly bactericidal effect against Vibrio cholerae.

In conclusion, this investigation reports that compound-1 isolated from the chloroform extract possesses antibacterial and antifungal activity and could be used in medicine after toxicity test.

REFERENCES
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