EVALUATION OF ANTIMICROBIAL POTENTIAL OF BAUHINIA BLAKEANA FLOWERSHTML Full Text
EVALUATION OF ANTIMICROBIAL POTENTIAL OF BAUHINIA BLAKEANA FLOWERS
D. Viji Saral Elezabeth and P. Ramachandran *
PG & Research Department of Chemistry, Nehru Memorial College, Puthanampatti, Tiruchirappalli - 621007, Tamil Nadu, India.
ABSTRACT: In this study, the phytochemicals and their antimicrobial potential of flowers of Bauhinia blakeana of Fabaceae Family were evaluated. The selected microbes for the antimicrobial study were E. coli, Enterobacter sp., Proteus sp., and Klebsiella sp. Ethanolic extract was prepared by continuous hot percolation method using Soxhlet extractor, and Antimicrobial activity was tested by Agar well diffusion method using Streptomycin as reference. The phytochemicals present in Bauhinia blakeana flowers were screened by standard method. The phytochemical evaluation revealed the presence of Alkaloids, Flavonoids, Glycosides, Terpenoids, Anthocyanins, Phytosterols, Tannins, Carbohydrates, Saponins, and Phenols. Among the phytochemicals, the Alkaloids and Flavonoids were showed strong results in the phytochemical screening. The Antimicrobial investigation revealed the extract of the flower showing significant Antimicrobial potential against the selected microbes. It showed maximum mean zone of inhibition of 12mm against Klebsiella sp. and a minimum of 7mm against Enterobacter sp. Further research on this plant flowers is to be emphasized.
Bauhinia blakeana, Phytochemicals, Agar well diffusion method, Antimicrobial, Soxhlet extraction
INTRODUCTION: Plant-derived drugs are playing a vital role in the modern medicinal world. The importance of the plants in the medicinal world is increased day-by-day significantly. The cause for their medicinal value is the presence of secondary metabolites such as alkaloids, terpenoids, tannins, flavonoid, and phenolic compounds. Bauhinia blakaena is an evergreen tree commonly present in India shown in Fig. 1. Phytochemical investigations showed that Acetone, Petroleum Ether and Methanolic extracts of Bauhinia blakaena’s flowers have antimicrobial activity against the Gram-positive and Gram-negative microbes.
FIG. 1: TREE AND FLOWERS OF BAUHINIA BLAKAENA
The present study was aimed to evaluate the phytochemicals and antimicrobial potential of Ethanolic extract of flowers of Bauhinia blakaena against the selected microbes.
MATERIAL AND METHODS:
Collection and Identification of Plant Material: The fresh part of the plant (flowers) was collected from Cholan Nagar, Tiruchirappalli District, Tamil Nadu State, India. The collection was under the supervision of a Horticulturist. This plant Bauhinia blakaena was authenticated by State Horticulture Farm, Mudhalaipatti (Village), Trichy Karur Road, Karur (District), Tamil Nadu.
The flower of Bauhinia blakaena was thoroughly washed and dried under shade for 4 weeks, segregated, pulverized by a mechanical mixer grinder to the fine powder before analysis. Fig. 2 was showing the pulverized plant material of Bauhinia blakaena flowers.
FIG. 2: PULVERISED PLANT MATERIAL OF BAUHINIA BLAKAENA
Preparation of Extracts: This sample (1:10 w/v) was successively extracted with Ethanol by hot continuous percolation method in Soxhlet apparatus shown in Fig. 3.
The extracts were concentrated by using a rotary vacuum evaporator and subjected to dryness to yield a crude residue. These residues were used for preliminary phytochemical screening of secondary metabolites and antimicrobial evaluation.
FIG. 3: SOXHLET EXTRACTION OF PLANT MATERIAL
Phytochemical Screening: The flower extracts of Bauhinia blakaena were analyzed for the presence of phytochemicals according to standard methods 1. The methods of phytochemical analysis are given in Table 1.
TABLE 1: PHYTOCHEMICAL SCREENING METHODS
|Phytochemicals||Name of the test||Procedure||Result|
|Alkaloids||Dragondroff’s test||Extract + Conc. H2SO4+reagent||Red – Orange precipitate|
|Mayer’s test||Extract + Conc. H2SO4+reagent||Yellow precipitate|
|Flavonoids||Lead acetate test||Extract + reagent||Yellow precipitate|
|Alkaline reagent test||Extract + NaOH + dil. HCl||Yellow to colorless|
|Carbohydrates||Molisch’s test||Extract + reagent||Violet ring formation|
|Fehling’s test||Extract + dil. HCl + NaOH + Heating with (A+B) reagent||Red precipitate|
|Saponins||Froth test||Extract + distilled H2O + shaking||Foam (1cm)|
|Foam test||Foam (10minutes)|
|Phytosterols||Libermann Burchard’s test||Extract + CHCl3 + (CH3CO)2O + Conc. H2SO4||Brown ring formation|
|Salkowski’s test||Extract + CHCl3 + Conc. H2SO4 + Shaking||Golden yellow colour formation|
|Legal’s test||Extract + Sodium Nitroprusside in pyridine + NaOH||Pink to blood red color|
|Terpenoids||Chloroform test||Extract + CHCl3 + Conc. H2SO4||Reddish brown ring formation|
|Steroids||Acetic anhydride test||Extract + (CH3CO)2O + Conc. H2SO4||Violet to bluish green|
|Proteins||Xanthoproteic test||Extract + Conc. HNO3||Yellow colour|
|Phenols||Ferric chloride test||Extract + FeCl3||Bluish black colour|
|Tannins||Ferric chloride test||Extract + distilled H2O + Heating + FeCl3||Dark green colour|
|Amino acids||Ninhydrin test||Extract + reagent||Purple, blue color|
|Anthocyanin||Sodium hydroxide test||Extract + NaOH||Bluish green color|
Microbial Strain: The microbial strains were collected from the Biotechnology Laboratory of Bishop Heber College, Tiruchirappalli (Ref. No.: BHC-BT-CTS04/2014/NMC) as pure cultures and used for the evaluation. The gram-positive and gram-negative bacteria’s namely E. coli, Proteus sp., Enterobacter sp. and Klebsiella sp. were taken for the test and they were cultured on Nutrient Agar (Hi-Media) Slants at 4 ºC.
Standard Antibiotic: Streptomycin (100µg/mL) was used as a reference standard against the pathogens.
Antibacterial Assay: The antibacterial activity assay of flower extracts was performed by agar well diffusion method. 20mL of sterile Muller Hinton agar (Hi-Media) was poured in sterile Petri dishes. The plates were allowed to solidify and used. 10mL of sterilized Muller Hinton agar medium (Seed Agar) was seeded with organisms (about 0.2mL according to 0.5 McFarland’s standard), in semi-hot conditions and was poured uniformly on the base agar. 8mm bores were made each equal distance from one another on the medium using sterile borer, and 100µL of different urine preparation were added to respective bore. The plates were incubated at 37 °C for 24 h, and the zone of inhibition was measured. For each test, three replicates were performed. Here an attempt was made to compare the antibacterial efficiency of flower extracts along with the activity of a standard antibiotic.
RESULT AND DISCUSSION: Fig. 4 was showing the result of the phytochemical evaluation of the plant extract.
FIG. 4: PHYTOCHEMICAL SCREENING OF PLANT MATERIAL
The results of preliminary phytochemical screening of acetone, petroleum ether, and methanol extracts of the flowers of Bauhinia blakeana are presented in Table 2.
TABLE 2: RESULT OF PHYTOCHEMICAL SCREENING OF ETHANOLIC EXTRACT OF BAUHINIA BLAKEANA FLOWERS
|Phytochemicals||Name of the test||Result|
|Flavonoids||Lead acetate test||+|
|Alkaline reagent test||+|
|Phytosterols||Libermann Burchard’s test||+|
|Cardiac Glycosides||Legal’s test||+|
|Steroids||Acetic anhydride test||-|
|Phenols||Ferric chloride test||+|
|Tannins||Ferric chloride test||+|
|Amino acids||Ninhydrin test||-|
|Anthocyanin||Sodium hydroxide test||+|
+: Indicates the presence of phytoconstituents
-: Indicates the absence of phytoconstituents
The phytochemical evaluation of ethanolic extracts of the flowers of Bauhinia blakeana revealed the presence of Alkaloids, Flavanoids, Terpenoids, Carbohydrates, Saponins, Phytosterols, Cardiac Glycosides, Phenols, Tannins, and Anthocyanins. Fig. 5 was showing a photograph of the mean zone of inhibition of Bauhinia blakeana against the selected microbes.
FIG. 5: ANTIMICROBIAL POTENTIAL OF ETHANOLIC OF PLANT MATERIAL AGAINST THE SELECTED MICROBES
The results of Antimicrobial activity of flower extracts of Bauhinia blakeana are furnished in Table 3.
TABLE 3: RESULT OF ZONE OF INHIBITION OF ANTIBACTERIAL ACTIVITY OF ETHANOLIC EXTRACTS OF BAUHINIA BLAKEANA FLOWER
|Name of the bacteria||E. coli||Enterobacter sp.||Proteus sp.||Klebsiella sp.|
|Mean Zone of Inhibition in mm||9||7||11||12|
The ethanolic extracts of Bauhinia blakeana flowers were exhibited different degrees of antimicrobial potential against the selected microbes. The antimicrobial potential against Klebsiella sp. (12mm) was more effective when compared with other microbes such as E.coli, Enterobacter sp., and Proteus sp.
CONCLUSION: It has been concluded that the flower extracts of the Bauhinia blakeana showed the presence of secondary metabolites such as Alkaloids, Flavanoids, Terpenoids, Carbohydrates, Saponins, Phytosterols, Cardiac Glycosides, Phenols, Tannins, and Anthocyanins. This investigation also concluded that the ethanolic extracts of the plant showed significant antimicrobial potential against the gram-positive and gram-negative bacteria’s by Agar well diffusion method. These studies are encouraged to isolate the active constituents of the plant.
ACKNOWLEDGEMENT: The Authors would wish to acknowledge the Management and Principal of Nehru Memorial College for providing research facilities and encouragement.
CONFLICT OF INTEREST: Nil
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How to cite this article:
Elezabeth DVS and Ramachandran P: Evaluation of antimicrobial potential of Bauhinia blakeana flowers. Int J Pharm Sci & Res 2014; 5(9): 3932-35. doi: 10.13040/IJPSR.0975-8232.5(9).3932-35.
All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
D. V. S. Elezabeth and P. Ramachandran *
PG & Research Department of Chemistry, Nehru Memorial College, Puthanampatti , Tiruchirappalli, Tamil Nadu, India.
14 March 2014
13 June 2014
01 July 2014
01 September 2014