Research Article ISSN: 0974-6943
Gulzar A. et al. / Journal of Pharmacy Research 2010, 3(11),2583-2584
Available online through www.jpronline.info Preliminary phytochemical and antimicrobial screening of leaves extract of Acacia catechu Willd Gulzar A. 1 *., Manjul P. S. 1 , Anita S. 1 , Upendra K. 1 , Yatendra K. 2 1
Kailash Institute of Pharmacy & Management, Gorakhpur, Utter Pradesh,,(India)
2
ITS Paramedical (Pharmacy) College, Muradnagar, Ghaziabad, Utter Pradesh,,(India)
Received on: 15-06-2010; Revised on: 18-08-2010; Accepted on:13-09-2010 ABSTRACT Acacia catechu Willd. (AC) is a potent medicinal plant in the traditional Indian medicinal systems. Preliminary phytochemical and antimicrobial investigations were carried out of the crude extract obtained from the leaves of AC. The presence of carbohydrates, steroids, alkaloids, glycosides, tannins, saponins, flavones, and phenolic compounds was indicated by the tests conducted. Antimicrobial activity of petroleum ether, ethanolic and ethanol:water (1:1) extracts of leaves of AC was evaluated by disc diffusion method against some pathogenic fungi and gram positive and negative bacteria. The Inhibition Zone Diameters of extracts were compared with standard Ketoconazole (10µg/disc) and Amikacin (30µg/disc) for antibacterial and antifungal activity respectively. Ethanolic extract was found to possess the broadest and potent antimicrobial activity and petroleum ether extract was found to be least active, while ethanol:water(1:1) extract playing an intermediate role.
Key words: : Acacia catechu Willd., Preliminary phytochemical & antimicrobial screening. INTRODUCTION Acacia catechu Willd. (AC) (Family: Fabaceae and subfamily: Mimosoideae) known as Black Khair. AC is commercially used to obtain Katha (a concentrated filtered extract) in North India. It is found widely distributed in Jammu, Punjab, Himachal Pradesh, Uttar Pradesh, Madhya Pradesh, Bihar, Andra Pradesh and Orrissa. AC grows slowly and matures to a height of about 12-15 meters. The species gets full foliage by June-July when it paints the environment and landscape so very beautifully. The leaves of AC are compound. The rachis branching from the mid-rib has 4 to 5 round prickles. The rachis is nearly 10 to 20 cm long and bears 20 to 60 pinnae each about 3 to 4 cm long. The tree flowers during June to October. Its inflorescence is pale yellow to cream colored. The fruit of is pod shaped. It is 5 to 7 cm long and 1 to 1.5 cm wide and shining brown in color. The sap wood of is large and yellowish white and heart wood is small and red in color. AC is useful in stomach problems like diarrhea, dysentery, colitis and gastric cancer. AC is used as mouthwash for mouth, gum and throat diseases like gingivitis, stomatitis. Cutch and katha is cooling, digestive and a very valuable astringent, especially in chronic diarrhea and dysentery, bleeding piles, uterine haemorrhages, leucorrhoea, gleet, atonic dyspepsia, chronic bronchitis, etc. The decoction of bark mixed with milk is taken to cure cold and cough [1-5]. The extracts of AC have been reported to have various pharmacological effects like antipyretic, anti-inflammatory, antidiarrhoeal, hypoglycaemic, hepatoprotective, antioxidant and antimicrobial activities [1, 3-7]. Main chemical constituents of Acacia catechu Willd are catechin, (-) epicatechin, epigallocatechin, epicatechin gallate, epigallocatechin gallete rocatechin, phloroglucin, protocatechuic acid, quercetin, poriferasterol glucosides, poriferasterol acyglucosides, lupenone, lupeol, procyanidin AC, kaempferol, dihydrokaemferol, taxifolin, (+)-afzelchin gum and mineral [3,8-14].
(U.P.), was authenticated by the plant taxonomist Dr. A.K. Sharma, Dept. of Botany, M.M.P.G. College, Modinagar (U.P.). A voucher specimen was deposited at the Dept. of Botany, M.M.P.G. College, Modinagar (U.P.). After authentication, fresh plant material (Leaves) was collected in bulk, washed under running tap water to remove adhering material, dried under shade and pulverized in a mechanical grinder. The coarse powder was pass through sieve no. 40.
An extensive search of the literature reveals no reports on the antimicrobial activity of AC leaves extract of the plant. Therefore an attempt was made to study the various phytochemical groups and antimicrobial potential of AC leaves extract against common human pathogenic organisms.
Antimicrobial screening The antimicrobial activities of plant extract i.e. petroleum ether extract, ethanolic extract and ethanol:water(1:1) extract were determined at different concentration (25µg/ml, 50µg/ml and 100µg/ml in dimethyl sulfoxide) against microbes by agar diffusion technique using a paper disc. Microbial cultures were firmly sweeped over the agar (nutrient agar medium) plate using sterile cotton swab to make uniform culture lawns. The saturated disc of different concentration of test compounds was placed on agar plates. Allowed the plates to stand at room temperature for 30 minute (pre-incubation). Inoculated plates were incubated at 37°C for 24 hr and 48hr for antibacterial and antifungal activities respectively. The inhibition zones were recorded with the help scale the result is summarized in Table 1. Amikacin (30µg/disc) and Ketoconazole (10µg/disc) was used as standard for antibacterial and antifungal activity respectively. Sterile dimethyl sulfoxide without plant extracts served as negat ive control [17, 18].
MATERIALS AND METHODS Plant material The leaves of AC was collected from Village Bageha, District Chitrakoot
*Corresponding author. Gulzar Alam Kailash Institute of Pharmacy & Management, Gorakhpur, Utter Pradesh,(India) Tel.: + 91-9696571822 E-mail:
[email protected]
Preparation of plant Extracts The extraction of 1 kg dried coarse powder of the AC leaves was carried out by cold maceration process using with petroleum ether (60-80oC), ethanol and ethanol: water (1:1) respectively for 7 days each. The petroleum ether (6080 oC), ethanol and ethanol: water (1:1) extracts were then filtered through muslin cloth and whatmann filter paper. The filtrates were concentrated under reduced pressure to obtain residues. Phytochemical Analysis Preliminary phytochemical screening of the leaves extracts of AC was performed as per standard procedure [15, 16]. which revealed that the presence of carbohydrates, steroids, alkaloids, glycosides, tannins, saponins, flavones, and phenolic compounds. Test microorganisms Pathogenic bacteria strains like Staphylococcus aureus (Gram positive), Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Salmonella typhi (Gram negative) and fungi like Candida albicans, Aspergillus niger were used for the screening of antimicrobial activity.
Journal of Pharmacy Research Vol.3.Issue 11.November 2010
2583-2584
Gulzar A. et al. / Journal of Pharmacy Research 2010, 3(11),2583-2584 Table 1 : Antimicrobial Activity of the Leaves Extracts of Acacia catechu Willd. Extract
Conc .
Petroleum ether extract Ethanolic extract Ethanol:water (1:1) extract Amikacin Ketoconazole Blank (DMSO)
25µg/ml 50µg/ml 100µg/ml 25µg/ml 50µg/ml 100µg/ml 25µg/ml 50µg/ml 100µg/ml 30µg/disc 10µg/disc ——-
Zone of inhibition (in mm) B1 B2 B3 B4
B5
F1
F2
09 15 16 10 13 15 07 12 15 22 ——-
09 12 13 10 13 17 08 12 14 22 ——-
09 11 12 10 13 16 09 11 12 —24 —-
10 12 11 11 14 15 08 10 13 —24 —-
12 14 15 11 12 18 08 11 14 22 ——-
08 09 10 09 10 11 08 10 11 22 ——-
08 10 11 10 11 15 09 11 16 22 ——-
20
Ext 1 (25µg/ml) Ext 1 (50µg/ml) Ext 1 (100µg/ml) Ketoconazole 10µg/disc Ext 2 (25µg/ml) Ext 2 (50µg/ml) Ext 2 (100µg/ml) Ketoconazole 10µg/disc
15
10
5
Ext 3 (25µg/ml) Ext 3 (50µg/ml) Ext 3 (100µg/ml) Ketoconazole 10µg/disc
0 er nig A.
ns ica alb C.
B1= Escherichia coli, B2=Staphylococcus aureus, B3=Pseudomonas aeruginosa, B4=Klebsiella pneumoniae, B5=Salmonella typhi, F1=Candida albicans, F2=Aspergillus niger.
25
Graph 2: Antifungal Activity of the Leaves Extracts of Acacia catechu Willd.
25 Ext 1 (25µg/ml)
20
Ext 1 (50µg/ml) Ext 1 (100µg/ml) 15
gation. Further work will emphasize the isolation and characterization of active principles responsible for antimicrobial activity of leaves extracts of Acacia catechu Willd.
Amikacin 30µg/disc Ext 2 (25µg/ml)
10
Ext 2 (50µg/ml) Ext 2 (100µg/ml) Amikacin 30µg/disc
5
Ext 3 (25µg/ml) Ext 3 (50µg/ml)
0 i ph S.ty
iae on em pn K.
osa gin eru P.a
s eu ur S.a
li co E.
Ext 3 (100µg/ml) Amikacin 30µg/disc
Graph 1: Antibacterial Activity of the Leaves Extracts of Acacia catechu Willd.
ACKNOWLEDGEMENTS The authors are thankful to I.T.S. Paramedical College, Muradnagar, Ghaziabad (U.P.), for providing bacterial strains like Staphylococcus aureus (Gram positive), Escherichia coli, Pseudomonas aeruginosa (Gram negative). I would like to place special thanks to I.T.S. Dental College, Muradnagar, Ghaziabad (U.P.), for providing bacterial strains like Klebsiella pneumoniae, and Salmonella typhi (Gram negative) and fungi like Candida albicans and Aspergillus niger as a gift sample for my research work. I would like to express my deepest gratitude to Dr. Y. Kumar, Director, I.T.S. Paramedical College, Muradnagar, Ghaziabad (U.P.), for providing me necessary facilities for carrying out research work. REFERENCES 1. 2.
RESULTS AND DISCUSSION The antimicrobial activity of the extracts at different concentration was determined. All the three extracts at different concentration exhibited antibacterial activity against all bacterial strains tested and antifungal activity against all fungal strain tested. Ethanolic extract exhibited comparably a high degree of activity than the other extracts (petroleum ether extract, ethanol:water(1:1) extract).In the present antimicrobial screening, ethanolic extract was found to possess the broadest and potent antimicrobial activity and petroleum ether extract was found to be least active, while ethanol:water(1:1) extract playing an intermediate role. The ethanolic extract was most effective against Staphylococcus aureus (Gram positive bacteria) with diameter of zone of inhibition 18.0mm (conc. 100µg/ ml) and was least effective against Pseudomonas aeruginosa (Gram negative bacteria) with diameter of zone of inhibition 11.0mm (conc. 100µg/ml). Amongst the Gram negative bacteria, the ethanolic extract showed high est activity against Salmonella typhi with diameter of zone of inhibition 17.0 mm. The ethanolic extract showed significant antifungal activity against Candida albicans with diameter of zone of inhibition 16.0mm (conc. 100µg/ml) and Aspergillus niger with diameter of zone of inhibition 15.0mm (conc. 100µg/ml).Phytochemical studies on AC leaves revealed the presence of carbohydrates, steroids, alkaloids, glycosides, tannins, saponins, flavones, and phenolic compounds. The antimicrobial activity of AC may be due to one/more group of above phytoconstituent (s).
3. 4. 5. 6.
7. 8.
9.
10. 11. 12. 13. 14.
15. 16.
CONCLUSION From the results, it can be concluded that the Acacia catechu Willd leaves extracts find use as broad-spectrum antimicrobial agent after extensive investi-
17. 18.
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Source of support: Nil, Conflict of interest: None Declared
Journal of Pharmacy Research Vol.3.Issue 11.November 2010
2583-2584
