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Molecular & Clinical Pharmacology 2012, 2(1) 34-43 Antibacterial and antidiarrhoeal activity of Alangium salviifolium Wang flowers Ronok Zahan1, M. Ashik Mosaddik1, Ranjan Kumar Barman 2, Mir Imam Ibne Wahed2, M. Ekramul Haque*,2. 1 Department of Pharmacy, BRAC University, Dhaka, Bangladesh 2 Department of Pharmacy, Rajshahi University, Bangladesh *Corresponding Author: [email protected] Abstract The present study was designed to investigate antibacterial and antidiarrhoeal pote- ntial of the extract/fraction of Alangium salviifolium flowers. The methanol extract (MEAS) along with some organic soluble fractions of the flower of Alangium salviifolium Wang were tested against six Gram-positive and six Gram-negative bacteria. Diethyl ether fraction (DEAS) was active against all tested microbial spe- cies and the highest activity was shown against Proteus sp with a zone of inhibition 12 ± 0.12 mm. The chloroform fraction (CAS) showed significant activity for all bacterial strains except Bacillus megatherium and Pseudomonas aeruginosa. The highest zone of inhibition was found against Escherichia coli (zone of inhibition 12±0.22 mm) for CAS. Among the all extract/fractions only pet ether fractions (PEAS) had no activity for all the test bacteria. CAS showed the maximum relative percentage inhibition against E. coli (34.21%) whereas; lowest relative percentage inhibition was found against Shigella flexneri (10.92%) for MEAS extract. The MEAS and CAS studied for antidiarrheal properties using castor oil and magnesi- um sulphate induced diarrheal model in mice. At the doses of 50 and 100 mg/kg body weight (MEAS) and 100 mg/kg b. wt. (CAS), the extract/fraction reduced the frequency and severity of diarrhea in test animals throughout the study period. Alt- ogether, these results suggest that the MEAS and CAS could be used as a potential antibacterial agent along with its antidiarrhoeal potentiality. Keywords: Alangium salvifolium; antibacterial Introduction The frequency of life-threatening infections caused by pathogenic microorganisms has increased worldwide and become a prime factor of morbidity and mortality in immune compromised patients in developing countries and many infectious microorganisms are resis- tant to synthetic drugs and hence an alternative therapy is very much needed (Okeke et al., 34 © 2012 Inforesights Publishing UK Zahan et al. 2005). Medicinal plants represent a rich source of antimicrobial agents. Plants are used medi- cinally in different countries and are a source of many potent and powerful drugs (Srivastava et al., 1996). Plants are rich in a wide variety of secondary metabolites such as tannins terpe- noids, alkaloids, flabonoids, etc, which showed wide range of in vitro antibacterial and antif- ungal activities (Dahanukar et al., 2000; Cowan., 1999) Traditional healers claim that their medicine is cheaper, more effective and impart least side effects as compaired to synthetic medicines. In developing countries, low income people such as farmers, people of small isol- ate villages and native communities use folk medicine for the treatment of common infecti- ons (Balandrin et al., 2006). The incidence of food-borne illnesses is still a major problem, even in developed cou- ntries. It has been estimated that 6–81 million cases of illnesses and up to 9000 deaths annua- lly were attributed to food-borne pathogens in the USA alone (Mead et al., 1999). Five patho- gens account for over 90% of the estimated food-related deaths: Salmonella (31%), Listeria (28%), Toxoplasma (21%), Norwalk-like viruses (7%), Campylobacter (5%) and Escherichia coli O157:H7 (3%). Oral rehydration therapy (ORT) remains the major treatment for diarr- hoea, although it does not reduce the volume or duration of diarrhea (Subbotina et al., 2003). Other treatment options include antibiotics and gut motility suppressing agents, which aim to reverse dehydration, shorten the length of illness and reduce the period of time an individual is infectious (Allen et al., 2003). Treatment with pharmacological agents that are pathogen- specific or that suppress severe symptoms would be of benefit to patients suffering from prolonged diarrhoea (Takahashi et al., 2001; Oi et al., 2002). Alangium salviifolium wang is a deciduous, rambling shrub or a tree belonging to the family Alangiaceae (Jubie et al., 2008). The different parts of this plant are used for a wide range of diseases. Root is used in diarrhoea, paralysis, piles and vomiting (Pandey et al., 2005). Root is also useful for external application in acute case of rheumatism, leprosy and inflammation (Anjaria et al., 2002). Antibacterial compound was isolated from the flower of Alangium salvifolium (Anjum et al., 2002). The plant has been reported for its anti-tubercula- r, anti-spasmodic and anti-cholinesterase activity (Warrier et al., 2005). Anti-Fertility activity of the stem Bark of Alangium salvifolium (Linn.F) Wang in Wistar female rats has also been reported (Murugan et al., 2000). Previous phytochemical investigation revealed that it is a rich source of alkaloids including ipecac alkaloid and benzopyridoquinolizidine alkaloids. It is also known to produce alangiside, a tetrahydroisoquinoline monoterpene glucoside (Itoh et al., 1992). Recent phytochemical studies of this plant resulted in the isolation of several flav- anoid, phenolic compound, irridoid glycosides and oxyoglucoside of some alcohol (Ramni et al., 2003). New alkaloid, ankorine was isolated from leaves (Jain et al 2002). Plant is also rich in tetrahydroisoquinoline monoterpene glycoside. e.g. alangiside-1 or ipecoside-2 whose structures are closely related to the ipecac alkaloid (Itoh et al 1994). In addition, considering the wide folk medicinal application of this plant, this work was set out in order to investigate the antimicrobial and antidiarrhoeal activity of extract and fractions of the flower of Alangiu- m salviifolium wang. Materials and methods Plant material © 2012 Inforesights Publishing UK 35 Molecular & Clinical Pharmacology 2012, 2(1) 34-43 The flowers of the Alangium salvifolium were collected from the adjoining area of Rajshahi University Campus, Bangladesh during February 2007 and were identified by Taxo- nomist, Department of Botany and University of Rajshahi, Bangladesh where a voucher spe- cimen number (Voucher No # 105) has been deposited. Preparation of extracts The flower material was shade-dried with occasional shifting and then powdered with a mechanical grinder, passing through sieve #40, and stored in a tight container. The powde- red flower (850gm) was taken in large glass bottle and extracted with methanol for 7 days. The procedure was repeated twice using same solvent system for next 3 days. The extract was filtered through filter paper. The filtrate obtained by repeated maceration was evaporated under reduced pressure at 40°c using Rotary evaporator. The net weight of dry extract was 5.5 gm. The dry plant extract (5.5gm) was suspended in water and fractionated in a conical flask using diethyl ether, Petroleum ether and chloroform solvent system. Each fraction furth- er evaporates using Rotary evaporator and then air dried to solid mass 250mg, 230 and 300mg, respectively. Preliminary Phytochemical Investigation The extract/fraction was subjected to qualitative chemical investigation for the identi- fication of different phytoconstituents like sterols, glycosides, saponins, carbohydrates, alkal- oids, flavonoids, tannins, proteins and tri-terpenoids (Yarnalkar., 1991). Test microorganisms Strains bacteria (Gram positive and Gram negative) were obtained from International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B). Bacillus anthracis ATCC 14321, Bacillus cereus ATCC 14579, Bacillus megaterium ATCC 13578, Bacillus subtilis A- TCC 6059, Streptococcus agalactiae ATCC 6123, Staphylococcus aureus ATCC 6538, Esc- herichia coli ATCC 25922, Pseudomonus aeruginosa ATCC 27853, Shigella flexneri ATCC 9221, shigella boydii ATCC 9234, Shigella dysenteriae ATCC 9361, Proteus sp. ATCC 93- 41, were used as test microorganism. All these bacterial species are recommended by ATCC for their susceptibility assay. The strains are maintained and tested on Nutrient Agar media (NA) for bacteria. Screening of antibacterial activity The methanol extract (MEAS) and soluble fraction of chlororform (CAS), diethyle- ther (DEAS) and petroleum ether (PEAS) of the flower of Alangium salvifolium were tested for antibacterial activity by disc diffusion method (Olurinola., 1996). For MEAS, CAS, DE- AS and PEAS, each (20mg) was dissolved in respective solvent (1 ml) to get a concentration of 200g/disc.