Antibacterial Properties of Ethanolic and Aqueous Extract of Banana Peels

Abstract

This work was done for the study of the antibacterial activity of the peel extract of Musa acuminata using ethanol, methanol, and an aqueous solvent of extraction. We have used the agar well diffusion method for this study and we know the antibacterial activity of the banana peel extract. Four concentration are used for this study ,these are 12.5 mg/ml ,25mg/ml,50mg/ml and 100mg/ml.

Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Klebsiella pneumoniae , Pseudomonas aeruginosa , and Salmonella enteritidis of this test microorganisms are used in this study to know the antibacterial activity of the peel extract of Musa acuminata. To study the phytochemical analysis of ethanol, methanol and aqueous extracts of the peel extract of Musa acuminata, flavonoids, terpenoids, quinines, and alkaloids are present in both ethanol, methanol, and aqueous extracts. So, phytochemical screening is revealed. From the result, we know that banana peels show antibacterial activity against these test microorganisms. Banana peels extract has great importance in public health.

The yellow banana peel has good antibacterial activity against Gram(+) and Gram(-) bacteria to replace the synthetic medicine in these diseases caused by these bacteria. From the result, we can know that Pseudomonas aeruginosa shows the maximum zone of inhibition (31.7+-2.0) in 100 mg/ml concentration. Staphylococcus aureus shows the minimum zone of inhibition (25.0+-2.0) in 100 mg/ml concentration of ethanol extract. In the lowest concentration (12.5 mg /ml), Bacillus subtilis shows the maximum zone of inhibition (19.0+-2.0) and Staphylococcus aureus shows the minimum zone of inhibition (15.5+-2.0) in ethanol extract.

Keywords: phytochemicals , antibacterial ,Musa acuminata ,banana peel.

Introduction

Musa acuminata (banana) is the most important food crop. It is developed in various countries. Banana is grown in 122 countries. And it is available in our country. Banana peel extract contains in many types of vitamins -vitamin B6, vitamin C, vitamin E, and malic acid. Mainly peel is the waste part of the various fruit. But the banana peel has antibacterial activity. In the case of commercial applications, peels might be due to their unknown benefit.

The potential application of banana peel depends on its chemical composition. Fatty acids are present in the banana peel and it responsible for the antibacterial activity of the peel extract. For this study, Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterititis of this bacterial culture are used to know the antibacterial activity. Methanol, ethanol and aqueous solvents are used.100 mg/ml,50mg/ml,25mg/ml and 12.5mg/ml concentration are done in aqueous, ethanol and methanol extract.

Material and method

Collection of plant material:

The fruit musa acuminata was selected for this study to know the antibacterial activity and phytochemical analysis. The fruit Musa acuminata was collected from our garden beside our home at kotalpara village. This specimen was identified in our microbiology department, Champadanga, Hooghly. After identifying this fruit, peels are collected from the banana fruit.

Preparation of the peel extract:

At first, the collecting peels are washed in well with the hot water and cold water. Then it is dried in air. The dried peels are grinded in the blender machine. The mixture is collected in a closed jar for long storage.

Preparation of aqueous extract:

5g of peel powder is added in the 25 ml of distilled water and made an aqueous solution. This solution is made in a conical flask. The conical flask is bounded with tissue paper and a rubber band. Then, some pore are done in the surface of the tissue paper which that air passes. The solution is kept in room temperature at 37℃ for 24 hours.

Preparation of ethanol extract:

5g of peels powder is added in 25 ml of ethanol and made into ethanol extract. This extract is made in a conical flask. The conical flask is bound with tissue paper and a rubber band. Some pore is done for air passes. Then it is kept in 37℃ room temperature for 24 hours.

Collection of test microorganisms :

The test microorganisms are Bacillus subtitles(MTCC-441), Staphylococcus aureus (MTCC-737), Klebsiella pneumonia (MTCC-432), Pseudomonas aeruginosa (MTCC-424), Escherichia coli(MTCC-443), Salmonella enterititis (MTCC - ) are collected. Those bacteria are grows on Moller Hinton agar and nutrients aged.

Preparation of different concentrations

12.5 mg/ml ,25 mg/ml ,50mg/ml and 100 mg/ml concentration are used for this study .1g of peel mixture is measured and it added in 10 ml of DMSO and made 100 mg/ml concentration. This is called a stock solution. For 50 mg /ml concentration,1 ml stock solution and 1 ml of DMSO are taken in a appended tube .0.50 ml stock solution is added in 1.50 ml DMSO and prepared 25 mg/ml concentration. For 12.5 mg/ml concentration ,0.25 ml stock is added with 1.75 ml DMSO .

Each concentration is kept in a 2ml volume of appends tube.6 nutrients agar plate are prepared and then it is inoculated with bacterial culture.3 plates are taken for 1 bacterial culture and another 3 plates are taken for another bacterial culture. Well, are done in each agar plate with the borar.100 mg/ml and 12.5 mg/ml concentrations are marked in two separate inoculated plates and given the concentration in the well from the particular tube. Another two inoculated plates are taken and then it is marked with 50 mg/ml and 25 mg/ml concentration and given concentration in the well from the particular tube. So, four plates are inoculated and given concentration. DMSO is given in another two plate.

Results obtained from this research work are summarized in the tables below.

Table -1 Antibacterial activity of the Musa acuminata in different bacteria for aqueous extract in different concentration

Concentration(mg/ml)100 mg/ml50 mg/ml25 mg/ml12.5 mg/mlEscherichia Coli27.25±2.022.25±2.018.25±2.017.25±2.0Bacillus Subtilis28.5±3.027.25±2.021.25±2.018.5±3.0Staphylococcus aureusL36.1±3.027.4±3.023.7±2.020.0±2.0Klebsiella pneumoniae32.3±2.028.2±3.023.0±2.019.5±2.0Pseudomonas aeruginosa34.5±2.031.7±2.026.0±2.021.2±3.0Salmonella enterititis29.0±2.026.2±3.023.1±2.019.4±3.0

The antibacterial activity of Musa acuminata shows the highest zone of inhibition against Pseudomonas aeruginous and the lowest zone of inhibition against Staphylococcus aureus.

Antibacterial activity of Musa acuminata shows the highest zone of inhibition against Pseudomonas aeruginosa and the lowest zone of inhibition against Staphylococcus aureus. Bacillus subtilis shows the maximum zone of inhibition (33.5±2.0) and Pseudomonusaeruginosa shows the minimum zone of inhibition (17.5±2.0).

Table-2 The antibacterial activity (inhibition zones in cm) of the peel extract of musa acuminate against different bactria for ethanolic extract in different concentration

Concentration(mg/ml)100 mg/ml50 mg/ml25 mg/ml12.5 mg/mlEscherichia Coli31.6±3.027.5±2.023.0±2.018.2±2.0Bacillus Subtilis33.5±2.030.1±2.026.4±2.021.7±2.0Staphylococcus aureus29.4±2.025.7±2.021.3±2.018.0±2.0Klebsiella pneumoniae32.9±2.030.7±3.026.1±2.021.6±2.0Pseudomonas aeruginosa27.6±2.022.4±2.020.4±2.017.5±2.0Salmonella enterititis30.0±2.027.4±3.024.5±2.018.7±3.0

Table-3 The antibacterial activity (inhibition zones in cm) of the peel extract of musa acuminate against different bactria for methanolic extract in different concentration

Concentration(mg/ml)100 mg/ml50 mg/ml25 mg/ml12.5 mg/mlEscherichia Coli26.1±2.022.3±2.020.4±2.016.6±2.0Bacillus Subtilis29.3±2.026.7±2.023.4±3.019.0±2.0Staphylococcus aureus25.0±2.021.2±2.018.7±2.015.5±2.0Klebsiella pneumoniae29.4±2.025.0±2.021.5±2.016.2±2.0Pseudomonas aeruginosa31.7±2.024.5±2.020.3±3.017.0±2.0Salmonella enterititis28.5±2.025.9±3.021.4±2.018.7±3.0

Pseudomonus aeruginosa shows the maximum zone of inhibition (31.7 ±2.0) and Staphylococcus aureus shows the minimum zone of inhibition (15.5±2.0).

Discussion

Banana is cheap fruit and readily available that is consumed by different people around the world.Because it has high nutritional properties. In recent times, it had been reported that these peels are not altogether useless as many of the bioactive plant components reside in them. Therefore, the project work was done to ascertain the antimicrobial efficacy of banana peels against clinical isolates.

From the phytochemical analysis of ethanol and aqueous extracts of banana peel, we know that flavonoids, terpenoids, quinines, and alkaloids are present in both ethanol and aqueous solvents, and tannins, saponins are absent in both solvents. Flavonoids are known as effective antimicrobial substances against a wide array of microorganisms. It is synthesized by plants in response to microbial attacks. Their activity is probably due to their ability to react with extracellular and soluble proteins and to complex with bacterial cell walls leading to the death of the bacteria. Tannins have various physiological effects like anti-irritant, antimicrobial and antiparasitic effects. Tannins-containing plants are used for treat nonspecific diarrhoea, inflammation of the mouth and throat, and slightly injured skins.

From this research work, we are found that which bacteria have good antibacterial activity. We are followed that Staphylococcus aureus is a susceptible bacteria species and it's zones of inhibition is 36.1+-3 in 100 mg/ml for aqueous extract. So, Lt shows good antibacterial activity in the highest concentration for aqueous extract. And lowest concentration (12.5 mg/ml) the zone of inhibition is 20.0+-2. Pseudomonas aeruginosa and Klebsiella pneumoniae are very sensitive in aqueous extract.

In the case of ethanol extract, Bacillus subtilis show good antibacterial activity in the highest concentration. So,the range of zone inhibition is 33.5+-2.0 in 100 mg/ml concentration. Klebsiella pneoniae and Escherichia coli is very sensitive in ethanol extract and the range of zone of inhibition is 32.9+-2.0 and 31.6+-3.0 in 100 concentration.

In the methanolic extract, Pseudomonas aeruginosa the range of zone inhibition is 31.7+-2.0 in 100 mg/ml. So, it shows good antibacterial activity. Bacillus subtilis and Klebsiella pneumoniae are very sensitive in methanol extract.The zone of inhibition are 29.3+-2.0 and 29.4 +-2.0 in 100 mg/ml.

Phytochemicals analysis

1. The peel extract of Musa acuminata was analyzed for alkaloids, tannins, glycosides, steroids, flavonoids, saponins, volatile oil, and resins using standard procedures.
2. Test for Glycosides:  To 1ml of the extract was taken, then 2 ml of acetic acid was added and then cooled in an ice bath at 4℃ .1ml of sulfuric acid is added in dropwise to this mixture. The formation of an oil layer on top of the solution that is indicated the presence of glycosides.
3. Test for alkaloids: 1ml of 1% HCL was added in 3ml of the extract. Then, the mixture is treated with a few drops of Meyer's reagent. If creamy white precipitation appears so, we can understand the presence of alkaloids.
4. Test for saponins:  5 drops of olive oil were added in 2 ml of plant extract and this mixture was shaken vigorously. A stable emulsion is formed that indicates the presence of saponins.
5. Test for tannins:  2 drops of 5% ferric chloride were added to 1 ml of the plant extract. A dirty-green precipitate is appeared which indicated the presence of tannins.
6. Test for flavonoids:  To 1 ml of the extract was added 3 drops of ammonia solution followed by 0.5 ml of concentrated HCL. The appearance of pale brown color in the mixture indicated the presence of flavonoids.
7. Test for steroids:  1ml of concentrated tetraoxosulphate (vi) acid was added in 1ml of the plant extract. A red coloration confirmed the presence of steroids.
8. Test for Resins:  To 5ml of the extract was added in 5 ml of copper acetate solution. The mixture was shaken vigorously and allowed to separate. Reddish-brown precipitation appears and it indicates the presence of resins.

Conclusion

In the case of the ethanolic and aqueous extract of banana peels, the antibacterial properties have been found to be considerably high in this research work. The test organism was highly resistant to antibiotics and they were found to be susceptible in the banana peels extract. Yellow banana peels have good antibacterial activity against both gram (+) and gram (-) and it is also known as ‘good antibacterial agent’. We are concluded that banana peel has good anti-bacterial activity in public health

Musa acuminata the wild species of banana is a plant of the tropical and subtropical regions. All parts of the plant including fruit, peels, leaves, roots are used in the treatment of many diseases in traditional medicine. The pharma ecological activities of musa acuminata include antioxidant, antidiabetic, anticancer, and antimicrobial especially anti-HIV activity. From the phytochemicals study, we can know the traditional use of different parts of musa acuminata in various diseases.

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