Chandrakant D. Shendkar et al.

/ Journal of Pharmacy Research 2012,5(1),102-103

Research Article ISSN: 0974-6943

Available online through www.jpronline.info

Comparative Evaluation of Achyranthes Aspera Linn. Parts by Antibacterial Activity

Chandrakant D. Shendkar 1 *, Pranav S. Chandrachood 2 , Sangita M. Lavate 1 , Bipinraj N. Kunchiraman3 , Nirmala R. Deshpande 1 . 1 Department of Chemistry, Yashwantrao Mohite College, Erandwane, Khothrud, Pune 411038, India. 2 Department of Chemistry, Dr. T. R. Ingle Research Laboratory, S. P. College, Tilak Road, Pune- 411030, India. 3 Department of Cell Biology, Rajiv Gandhi Institute of Biotechnology, Bharati Vidyapeeth University, Pune 411046, India.

Received on:20-09-2011; Revised on: 15-10-2011; Accepted on:10-12-2011 ABSTRACT

Achyranthes Aspera Linn.belongs to the family amaranthaceae. In Ayurvedic system of medicine various parts of this plant are employed to treat various type of infectious diseases. In this study, acetone extracts of leaves, stem and roots, prepared at room temp, were employed. The extracts were studied against Gram-positive and Gram-negative bacterial strains such as bacillus subtilis, staphylococcus aureus, salmonella typhimurium, escherichia coli, pseudomonas aeruginosa, salmonella abony. Among these extracts, leaves extract exhibit significant antibacterial activity against all tested bacterial strains while stem demonstrated feeble activity. All extracts displayed activity against almost all tested bacterial strains. Thus, as reported in Ayurveda, such extracts can be an alternative potent therapy for infectious diseases caused by these tested pathogenic bacterial strains. KEYWORDS: Antibacterial activity, Achyranthes Aspera Linn., Acetone Extracts, Disc diffusion method. INTRODUCTION: Plants are the richest resource of drugs of traditional systems of medicine, modern medicines, folk medicines, pharmaceutical intermediates and chemical entities for synthetic drugs [1] A wide range of medicinal plant parts extract are used as raw drugs and they possess varied medicinal properties. Microorganisms have developed resistance to many antibiotics and this has created vast clinical inconvenience in the treatment of infectious diseases [2].The increase in microorganisms resistance to antibiotics, the use of antimicrobial drugs forced scientists to search for new antimicrobial substances from various sources including medicinal plants [3]. The trend of using natural products has increased and the active plant extracts are frequently used for new drug discoveries and for the presence of antimicrobial substances [4]. Achyranthes aspera Linn. belongs to the family amaranthaceae is one of the important medicinal plant. It is used as traditional healers in the treatment of fever, especially malaria fever, dysentery, asthma, hypertension and diabetes [5,6] . The chloroform and ethanol root extracts of the A. aspera are reported to have anti-implantation and abortifacient activity [7,8] .The ethanol extract of the root posses spermicidal activity[9]. The literature survey indicate that aqueous and methanolic extracts of the whole plant have hypoglycaemic effect [10]. Roots are used as astringents to wounds, in abdominal tumor and stomach pain [11] . The stem bark shows abortifacient activity in the rat [12] . Leaf extracts were reported to posses thyroidstimulating and antiperoxidative properties [13].The present work is carried out to evaluate the efficacy of leaves, stem and roots extracts of this plant . MATERIALS AND METHODS: The plant material was collected from the Purander district of Pune,

Maharashtra, India. It was authenticated from Botanical survey of India, Pune, Maharashtra, India. Air shade dried and powdered material of leaves, stem and roots ( 10gm ) of each was extracted by using acetone (70 ml) separately by soaking it for 24 hours at room temperature. The solvent was collected under reduced pressure to obtain crude extracts of leaves, stem and roots. Antibacterial studies were carried out against six bacterial strains ( salmonella typhimurium, escherichia coli, psedomonas aeruginosa, salmonella abony, bacillus subtilis, staphylococcus arueus). The paper disc diffusion method was employed. Samples of each extracts (40 mg) were dissolved in respective solvent ( 1 ml ). Sterile Whatman filter paper discs (5 mm diameter) were impregnated with 10 L of these solvent extracts (400g /disc). The nutrient agar medium was prepared and transferred to the sterile petridishes in such a way to keep a uniform depth of approximately 4mm in sterile area. The 100l of test organism cultured in nutrient broth media was spread with a sterile spreader on the surface of solid nutrient agar media. The sterile discs impregnated with different extracts were placed on agar plates. The bacterial plates were incubated at 37 0.1 C for 24 hours. After incubation, all the plates were observed for zones of inhibition and the diameters of these zones were measured in millimeters. All tests were performed under sterile conditions. Ampicillin ( 1mg/disc ) was employed as positive control. RESULTS AND DISCUSSION: The acetone extracts of leaves, stem and roots of this plant showed varying degree of antibacterial activities against the tested bacterial strains . Among these, acetone leaves extract is found to be the most effective against salmonella typhimurium, escherichia coli, staphylococcus arueus, bacillus subtilis, psedomonas aeruginosa, salmonella abony. Root extracts showed activity against salmonella typhimurium, escherichia coli, bacillus subtilis, staphylococcus arueus, psedomonas aeruginosa. Stem extracts showed activity against salmonella typhimurium , escherichia coli, bacillus subtilis, staphylococcus arueus, psedomonas aeruginosa.

*Corresponding author.
Chandrakant D. Shendkar Department of Chemistry, Yashwantrao Mohite College, Erandwane,Khothrud, Pune 411038, India.

Journal of Pharmacy Research Vol.5 Issue 1.January 2012

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Chandrakant D. Shendkar et al. / Journal of Pharmacy Research 2012,5(1),102-103

The results of the antibacterial assay of acetone extracts of leaves, stem and roots of the Achyranthes aspera Linn. are presented in (Table 1). Table 1: Zone of inhibition of Acetone extracts of leaves, stem and roots
Test bacteria Zone of inhibition(mm)a Acetone Acetone Acetone leaves roots stem extract extract extract 400g /disc 13 10 9 9 7 6 11 8 7 8 8 10 7 8 6 7 -

CONCLUSION: The results obtained in the present study suggest that the acetone extracts of leaves, stem and roots of A. aspera reveals a significant scope to develop a novel broad spectrum of antibacterial drug formulation. This plant extracts would help for development of a new alternative medicine system. ACKNOWLEDGEMENTS: The authors are thankful to the Principal, Yashwantrao Mohite College and Rajiv Gandhi Institute of Biotechnology, Bharati Vidyapeeth University, Pune, Maharashtra, India for providing laboratory facilities and space for work. REFERENCES:
1. 2. 3. Hammer KA, Carson CF, Riley TV, Antimicrobial activity of essential oils and other plant extracts, J. Appl. Microbiol, 1999; 86(6) : 985. Davis J, Inactivation of antibiotics and the dissemination of resistance genes, Science, 1994; 264: 375- 382. Karaman I, Sahin P, Gulluce M, Oguten H, Songul M, Adiguzed A, Antimicrobial activity of aqueous and methanol extracts of Juniperus oxycedrus L, Journal of Ethnopharmacolology, 2003; 85(2-3) : 231235. Das S, Das S, Pal S, Mujib A, Dey S, Biotechnology of medicinal plants- Recent advances and potential, 1st Ed., Vol 2, UK 992 Publications, Hyderabad, 1999: 126139. Girach RD, Khan ASA, Ethananomedicinal uses of Achyranthes apera leaves in Orissa (India),Int J. Pharmacognosy,1992 ;30:113-115. Tang W, Eisenbrand G, Achyranthus bidentata Bl. in Chinese Drugs of Plant Origin, Springer-Verlag.,Berlin,1992:13-17. Bhom KH, Liersch R, Haensel R, Keller K, Rimpler H, Schneider (Eds.).G, Hagers Handbchder Pharmazeutischen Praxis Springer Verlag, Berlin, 1992; 5: 54-59. Vasudeva N, Sharma SK, Estrogenic and pregnancy interceptory effects of Achyranthes aspera Linn.Root, J.Ethanopharmacol, 2006; 107: 179. Paul D, Bera S, Jana D, Maiti R, Ghosh D, In Vitro determination of the contraceptive spermicidal activity of a composite extract of Achyranthes aspera and Stephania hernandifolia on human semen, Contraception, 2006; 73:284-288. Mohammad SA, Javed I,evaluation of hypoglycemic effect of Achyranthus aspera in normal and alloxan diabetic rabbit, Journal of Ethanopharmacology, 1991; 31(1):49-57. Ghani A, Medicinal Plant of Bangladesh with Chemical Constituents and Uses. 2nd ed. Asiatic Society of Bangladesh, Dhaka, 2003: 71-72. Bhattarai N, Folk herbal remedies for gynaecological complaints in Central Nepal, Int J Pharmacog, 1994;32(1):13-26. Tahiliani P, Kar A, Achyranthes aspera elevates thyroid hormone level and decrease hepatic lipid peroxidation in male rats, J Ethanopharmacol, 2000;71: 527-532.

Ampicillin 1mg/disc 16 12 11 14 10 8

Salmonella typhimurium Bacillus subtilis Escherichia coli Staphylococcus arueus Psedomonas aeruginosa Salmonella abony

a. Zone of inhibition including the diameter of Whatman filter paper disc ( 5 mm)

It must be noted that the largest zone of inhibition (13mm) is obtained with leaves extract against salmonella typhimurium. Therefore the MIC of this extract is determined by testing different concentrations of acetone leaves extract against salmonella typhimurium by disc diffusion method. The antibacterial assay of the leaves extract of A.aspera against salmonella typhimurium at different concentrations (300 g, 200 g, 100 g, 50 g, 25 g, 20 g/disc) are tested by disc diffusion method and are presented (Table 2). All zones of inhibition are represented including the diameter of disc. Table 2: Zones of inhibition of leaves extract against salmonella typhimurium at different concentrations :
Test bacteria 300 g/disc Salmonella typhimurium 12 Zone of inhibition(mm)a Acetone leaves extract 200 100 50 25 g/disc g/disc g/disc g/disc 10 9 9 7 Ampicillin 20 g/disc 1mg/disc

4.

5. 6. 7.

8.

16

9.

a. Zone of inhibition including the diameter of Whatman filter paper disc ( 5 mm)

The antibacterial activity of leaves extract against salmonella typhimurium showed significant reduction in bacterial growth in terms of zone of inhibition. The zone of inhibition decreases on decreasing the concentration of extract. This shows the concentration dependent activity. The MIC (Minimum Inhibitory concentration) is the lowest concentration of the compound at which the tested bacteria does not demonstrate visible growth at 20 g/disc. The result reveals that the MIC of leaves extract of A.aspera against salmonella typhimurium is 25 g/disc.

10.

11. 12. 13.

Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.5 Issue 1.January 2012

102-103