The bark of Bauhinia racemosa are reported to have great medicinal value. Phytochemical screening of the plant bark reveals the presence of carbohydrates, alkaloids, steroids and tannins. the methanol, ethanol, aqueous, acetone and petroleum ether extracts of bark of B. racemosa Lamk. prepared and antimicrobial activity were studied by agar well diffusion method against enteric bacterial pathogens such as E. coli, S. aureus, B. subtilis, P. aeruginosa and fungi A. niger and C.albicans. The methanol extracts had wide range of antimicrobial activity against enteric microbes than ethanol extracts, where as ethanol extract were slightly higher antibacterial activity than aqueous extract. Antimicrobial activity of various extracts of bark of B. racemosa was carried in attempt to develop a new pharmaceutical drug from natural origin for prevention of enteric infection

Chavan, R T

Kadam, A S

Current Botany

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Current Botany 2012, 3(4): 31-33 ISSN: 2220-4822 Available Online: http://currentbotany.org/    Preliminary phytochemistry and antimicrobial activity of bark of Bauhinia racemosa Lamk.  R. T. Chavan and A. S. Kadam   Department of Botany, DSM’S ACS College Jintur Dist. Parbhani -431509 (M.S.), India  Abstract  The bark of Bauhinia racemosa are reported to have great medicinal value. Phytochemical screening of the plant bark reveals the presence of carbohydrates, alkaloids, steroids and tannins. the methanol, ethanol, aqueous, acetone and petroleum ether extracts of bark of B. racemosa Lamk. prepared and antimicrobial activity were studied by agar well diffusion method against enteric bacterial pathogens such as E. coli, S. aureus, B. subtilis, P. aeruginosa and fungi A. niger and C.albicans. The methanol extracts had wide range of antimicrobial activity against enteric microbes than ethanol extracts, where as ethanol extract were slightly higher antibacterial activity than aqueous extract. Antimicrobial activity of various extracts of bark of B. racemosa was carried in attempt to develop a new pharmaceutical drug from natural origin for prevention of enteric infection.  Keywords: Bauhinia racemosa, antimicrobial activity, bark. INTRODUCTION       The use of plant compounds for pharmaceutical purposes has gradually increased in India 80% of individuals from developed countries use traditional medicine which involves compounds derived from medicinal plants [1] . The use of plant extracts and phytochemical both with known antimicrobial properties can be of great significance in therapeutic treatments [2]. Hence studies involving the use plants as therapeutic agents should be emphasized, especially those related to the control of antibiotic resistant microbes. The plant B.racemosa Lamk. belonging to the Caesalpiniaceae family. It occurs frequently in India, Ceylon, China and Timor. The stem bark of the plant is an astringent and is used in the treatment of headache, fever, skin diseases, tumors, blood diseases, dysentery and diarrhea. The fresh flower buds of the plant showed antiulcer activity [3 and 4].      Therefore the review revealed that the bark of B. racemosa Lamk. were used in various metabolic disorders but for their antimicrobial properties were not demonstrated. Hence attempt was made to find out the antibacterial properties of bark of B.racemosa Lamk. against enteric bacterial pathogens.   MATERIALS AND METHODS  Plant material        Fresh bark of B.racemosa Lamk. were collected from the natural population growing in different areas of Parbhani district of Maharashtra. Plant was identified and authenticated from the herbarium of Deptt. of Botany Dr. B.A.M. University, Aurangabad, flora of Marathawada [5] and encyclopida of Indian medicinal plants. Preparation of extracts       The fresh bark was dried under shed at room temperature and then powdered with a grinder and stored in air tight container. A soxhlet apparatus was used for extraction of antimicrobial active compounds from the powder. The dried powder of the bark of the plant (20 gm) was successively extracted using methanol, ethanol, aqueous, acetone and petroleum ether solvents respectively. The collected extracts were concentrated by evaporation under room temperature and used for the study.  Phytochemical screening       Different extracts were screened for the presence of alkaloids, glycosides, flavonoids, saponins, tannins, sterioids, resins, phytobatanine, oil and fats by using standard protocol [6 and 7].   Test microorganisms       Bacterial and fungal strain used for testing included authentic pure cultures of human pathogenic bacteria like. Staphylococcus aereus (SRTCC1073), Bacillus subtilis (SRTCC1091) and two are gram negative viz. Pseudomonas aeruginosa (SRTCC708), Escherichia coli (SRTCC3260). Two species of fungi viz. Aspergilus niger (SRTCC1073), Candida albicans (SRTCC3971).These were obtained from the school of life sciences, S.R.T.M. university, Nanded (M.S.).  Preparation of test organisms suspension        The test organisms were maintained on slants of medium containing nutrient agar (2.5 gm/ 10ml)  and sub cultured once a week. The slants incubated at 370C for 24 hrs and stored under refrigeration. The inoculums was 1x 108 cells/ml.  Antimicrobial activity       The in vitro antimicrobial activity of different bark extracts of B. racemosa Lamk. was determined by agar well diffusion method [8] .   Received: July 12, 2012; Revised: Aug 18, 2012; Accepted: Oct 10, 2012.  *Corresponding Author R.T.Chavan  Department of Botany, DSM’S ACS College Jintur Dist. Parbhani -431509  (M.S.), India  Email: rtcbotany@gmail.com   Chavan and Kadam   32The plant extracts were dissolved in distilled water at concentrated 50 µg/ ml streptomycin was used as reference antibiotic. Each plate was inoculated with 20µl microbial suspension having concentration 1x 108 cells/ml.  The 0.1 ml extract was added to each well. The plates containing bacteria were incubated at 370C for 24 hrs and those containing fungi were incubated at 250C for 7 days. Positive antimicrobial activity was based on growth inhibition zone and compared with standard drug [9]. The dimeter of zone of inhibition surroundings each of the well was recorded. All the experiments were performed in triplicate.  Statistical analysis       Results were expressed as mean ± S.D. statistical significance was determined using analysis of students t- test.  RESULTS AND DISCUSSION       Indian systems of medicine such as ayurveda and siddha uses majority of the crude drugs that are of plant origin. It is necessary that standards have to be laid down to control and check the identity of the plant an ascertain its quality before use. A detailed pharmacognostic evaluation there fore is highly essential perquisite [10].       The phytochemical investigation of the various solvent extract of bark of B. racemosa Lamk. Presented in table 1. the results revealed that the bark of B. racemosa Lamk. Show the presence of alkaloids,glycosides, carbohydrates, saponins, flavonoids, triterpenoids, anthroqunonine, phytobatalanine are present in methanol, ethanol and aqueous extracts. The result of the antimicrobial activity of the different extracts of bark of B. racemosa Lamk. are presented in table 2. In the present investigation methanol and aqueous extracts of bark showed higher antibacterial activity against the test organisms which was greater than the standard reference antibiotic streptomycin. The methanolic extract of bark also found more inhibitory activity on gram negative and gram positive bacteria E. coli, P.aeruginosa, S. aureus and B.subtilis. The various worker have already shown that gram positive bacteria are more susceptible towards plants extracts as compared to gram negative [11]. The difference in the activity may be due to the different secondary metabolites present in extracts. The gram negative activity bacteria P. aeruginosa shows the higher activity in both ethanol and water extracts the antimicrobial activity of the ethanol (23mm) was higher than that of water extract (19mm). Kumar (1999) [12] reported that methanol extract of B. racemosa Lamk. root bark possess significant inhibition effect against a broad range of microbes in dysenteriae, vibrio cholera, S. aureus, S.pnemonia, M. luteus, C. albicans and A. niger. Table 1.Phytochemical analysis of bark of Bauhinia racemosa Lamk.  Sr. no. Phyto Constituents Chemical tests B. racemosa bark extract P.E Met Eth Aq Ac 1 Alkaloids 1. Mayer’s test - - + - - 2. Dragendroff’s test - + - + - 3. Wagner’s test - - + - - 4. Hagers test - - - - - 2 Carbohydrates 1. Molisch’s test - + + + - 2. Benedicts test - - - - - 3. Fehling’s test - - - - - 3. Glycosides 1. Modified Borntragers - + + +  2. Legal test - + - +  4 Saponins 1. Foam test - + - + - 2. Froth test - - - - - 5 Triterpenes 1. Salkowski test - - + + - 2. Libermann Burchard - - - - + 3. Tschugajew test - - - - - 6.  Fats & Oil 1. Stain test - - - - - 7 Tannins 1. Alkaline Reagent - + - + - 8 Flavanoids 1. Gelatin test - + + + - 2. Lead acetate test - + + +  3. Shinoda test - + + + + 4. Zn-Hcl reduction  - + + +  9 Photobatalin  - + + - + 10 Anthraqunonines  - + + - +  Table 2. In vitro antimicrobial activity of bark of Bauhinia racemosa Lamk.  Microorganisms Diameter of Zone of inhibition in mm of different extracts of bark (2mg/ml) Mean± S.D. Standards reference antibiotic (Streptomycine) Bacteria Methanol Ethanol Aqueous Pet. ether Acetone  Escherichia coli 15±1.23 14±1.23 14.3±1.11 12±2.12 11±2.11 14±1.11 Staphylococcus aureus 14.8±1.11 14±2.12 15±1.11 11±1.12 12±1.21 14.3±1.11 Bacillius subtilis 15±1.15 13±1.21 15±1.43 13±1.23 13±1.32 13±1.12 P.aeruginosa 15±2.23 14±2.31 14±1.11 14±1.21 14±1.23 14±1.14 Fungi  Aspergillus niger 13±1.13 12±1.23 12±1.11 12±2.12 12±2.31 15±1.15 Candida albicans 12±0.12 12±3.33 13±1.24 11±1.11 12±1.11 14±1.13   Current Botany 2012, 3(4): 31-33  33 REFERENCES  [1] Chopra R. N., Nayar S. L., Chopra I. C. 1986. Glossary of Indian Medicinal Plants (including the supplement). Council of Scientific and Industrial Research, New Delhi.  [2] Kirtikar, K. R. and Basu, B. D. 1987. Indian Medicinal Plants. 2nd Edition. International Book Distribution Vol. III. Dehradun. Pp- 1664-1666. [3] El-Khatiba, A. S. and Khaleel, A. E. 1995. Evaluation of some Pharmocological Properties of Different Extracts of Bauhinia racemosa Leaf and Bassia muricata Whole Plant. Bulletin of the Faculty of Pharmacy, Cairo University. 33: 59-65. [4] El Hossary, G. A., Selim, M. A. Sayad, A. E. and Khaleel, A. E. 2000. Study of the Flavonoid Content of Bassia muricata and Bauhinia racemosa. Bulletin of the Faculty of Pharmacy, Cairo University. 38: 93-97. [5] Naik V.N. 1998. Flora of Marathwada, Amrut Prakashan, Aurangabad. [6] Trease, G.E. and Evans, W.C. 1989. Pharmcognsy 11th edn. Brailliar Tiridel Can. Macmillian publishers.  [7] Sofowara, A. 1982. Medicinal plants and traditional medicine in Africa Spectrum Books Ltd. Ibadan, Nigeria.Pp-289. [8] Perez, C. and Paul, M and Bezique, P. 1990. An antibiotic assay by the agar well diffusion method. Alta Biomed, Group Experiences.15:113. [9] Perez, G.R.M., Avila, J.G., Zavala, M.A., Perez, G.S. and Perez, G.S. 1996. IN Vitro antimicrobial activity of Loeselia Mexicana and Croton ehrenbergii. Phytomedicine. 3:186. [10] Ramana, P. 2007. Quality control and standardization of herbal drugs. A tool between the volume of antimicrobial consumption in human communities and the frequency of resistance. Proc Nati Acad Sci USA.96: 1152-6. [11] Parekh, J. and Chanda, S. 2006. Screening of some Indian medicinal plants for antibacterial activity. Indian Journal Pharm Sci.68: 835-838. [12] Kumar, K.A. 1999. Antioxidant and antimicrobial activities of Bauhinia racemosa L. stem bark. Brazilian Journal..3: 121-122.   

Preliminary phytochemistry and antimicrobial activity of bark of Bauhinia racemosa Lamk.

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Preliminary phytochemistry and antimicrobial activity of bark of Bauhinia racemosa Lamk.

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