Phytochemical Analysis and Antimicrobial Activity of Ethonolic Leaf Extract of Ficus racemosa Linn

 

T. Thilagavathi*,  G. Kathiravan

Department of Biotechnology, Vels University Pallavaram, Chennai, India

*Corresponding Author E-mail: g_kathir72@gmail.com

 

ABSTRACT:

The present study was conducted to analyze the phytochemicals like tannins, Saponins, Flavonoids, terpenoids, steroids, glycosides, reducing sugar, resins and anthocyanins in the ethanolic extract of the leaf of Ficus racemosa by standard procedure. The antimicrobial activity of the plant extract was evaluated against the three bacteria and three fungi at different concentrations by using disc diffusion and agar well diffusion methods. The plant sample was found to be bacteriostatic and fungistatic in action, thus can be used as a source of antibiotic agents for drug development that can be used in the control of these bacterial and fungal species.

 

KEYWORDS: Tannin, Flavonoids, Antibacterial, Antifungal, Ethanolic extract, Ficus racemosa.

 

 


INTRODUCTION:

Medicinal plants contain some active compounds which produce definite physiological action on the human body.1Many of the natural compound in plants of medicinal value offer us a new source of drug which have been utilized effectively in traditional medicine. Phytochemicals are the natural bioactive compounds of plant origin. They are found in all parts the plant body and the plant parts may differ from one part to another.2 The medicinal plant of phytochemicals with known antimicrobial properties can be of great significance in therapeutic treatments.3 In developing countries, it is surveyed that about 80% of the population rely on traditional medicine for their primary health care.4Since the time immemorial, different parts of medicinal plant have been used to cure specific ailments in India. Natural antimicrobials can be derived from many natural sources of plant; animal tissue and microorganism.5A large number of medicinal plants are used in different formulations for the treatment of various diseases caused by microbes. Ficusracemosa. Linn (Family: Moracea) is used in traditional medicine for the treatment of several disorders.

 

It is one of the herbs used in all ancient scripture of Ayurvedha, Siddha, Unani, and Homeopathy.6 Apart from the usage in traditional medicine, scientific studies mention F. racemosa to posses various biological effects such as heaptoprotective, chemopreventive, 7antidiabetic, anti-inflammatory8 antipyretic, antitussive, and antidiruetic. Different parts of F. racemosa were evaluating for blood sugar lowering activity in sterptozotocin induced diabetic rats.9The important phytochemicals are tannins, saponins, flavonoied, terpenoied, steroid, glycosides, anthocyonins, and resins, etc. These compounds are synthesized by primary or rather secondary metabolism of living organism serving as potential sources of new compounds of therapeutic and lead compounds in the drug development. The pharmacological studies mention that phytochemicals can prevent colorectal cancer and other cancers.10In the present work, qualitative phytochemical analysiswas carried out onthe F. racemosa plant extract and evaluate the antimicrobial activity in three bacteria and three fungi at different concentration by using disc diffusion methods.

 

MATERIALS AND METHODS:

Collections of plant materials:

Fresh plantparts of F.racemosa were obtained from Nagapattinam district Mayiladuthurai. The leaves were washed with sterile water and shade dried until the entire water molecule evaporated and plants become well dried for grinding.  After drying the plant samples were ground well and made into fine powder stored in airtight containers.

 

Preparation of plant extract:

10 gm of dried finely powdered plant material was taken in a beaker and 50 ml of ethanol was added. The mixture was keptin normal temperature at an orbital shaker for 24 hrs. Then the ethonolic extract was filtered through filter paper and the filtrate was used for the phytochemical analysis.

 

Phytochemical analysis:

The ethonolic leaf extract was carried out to analysis the phytochemical screening by using the following standard methods.11

 

Test for tannins (Ferric chloride test):

Three drops of 10% FeCl3 solution were mixed with 2ml of F. racemosa plant extract and Silver nanoparticles. The appearance of blackish green coloration indicates the presence of tannins.

 

Test for Saponin:

About 2g of leaf powdered and powder of silver nanoparticle is boiled with 20 ml of distilled water in a water bath and filtered. 10 ml of the filtered sample is mixed with 5 ml of distilled water in a test tube and shaken vigorously to obtain a stable persistent forth. The frothing is then mixed with 3 drops of olive oil and observed in the formation of emulsion which indicates the presence of Saponin.

 

Test for Flavonied:

The two samples were used for the following test: three ml of each sample was mixed with 4ml of 1% aluminium chloride in methanol in a test tube and the color was observed. Formation of yellow color indicated the presence of flavonols, flavones and chalcones.

 

Three ml of each sample was mixed with 4ml of 1% potassium hydroxide in a test tube and the color was observed a dark yellow color indicate the presence of flavonied. 

 

Test for terpenoids: (Salkowski test):

Around five ml of each sample is mixed with 2 ml of chloroform in a test tube and 3ml of concentrated sulfuric acid is carefully added. An interface with a reddish brown coloration is formed if terpenoied is present.

 

Test for Steroids: (Salkowski test):

0.5games of the each sample were mixed with 2ml of acetic anhydride followed by 2ml of sulfuric acid. The color changed from violet to blue or green in some sample indicates the presence of steroid

 

Test for glycosides: (Keller killiani test):

To 1 ml of each sample a few drops of glacial acetic acid and ferric chloride and 3-4 drops of concentrated sulfuric acid were added. The appearance of blue-green color indicates the presence of glycosides.

 

Test for reducing sugar: (Fehling’s test):

With 0.5 ml of each sample was treated with one ml of water and 5-8 drops of Fehling’s solution were added to the test tube hot and observed for brick red precipitate.

 

Test for Resins:

One ml of each samplewas treated with a few drops of acetic anhydride solution followed by one ml of Conc. H2So4.  Resins give coloration ranging from orange to yellow.

 

Anthocyanins:

2 ml of each sample, two ml of 2NHCl and ammonium were added and appearance of pink coloration, it formed red colors and turns the blue violet it shows the positive result.

 

Assay for antimicrobial activity:

Test microorganism

Escharicha coli, Psuedomonas auerogenus, and staphylococcus aureus, were used to determine the antibacterial activity while Aspergillus flavus, Rhizopus, and Rhizoctonia solani species were used to assess the antifungal activity.

 

Antimicrobial Activity using the disc diffusion method:

The Antibacterial activity of the F. racemosa plant extract was performed by disc diffusion method.12 About 20 ml of sterile molten Muller Hinton agar was poured into the sterile petri plate, after solidification 100 µl overnight cultures of human pathogen were swabbed on the respective plates. The disc was kept over the agar plate using sterile forceps at various concentrations (50, 100  and 150µl). The plates were incubated for 37° C for 24 hrs. After incubation the diameter of inhibitory zones formed around each disc was measured (mm) recorded. The antibacterial assay of plant samples against all microorganisms tested was performed in triplicates.

 

Antifungal activity using disc diffusion method:

Antifungal activity of F. racemosa plant extract was determined by disc diffusion method on potato Dextrose agar (PDA) medium.13 the potato dextrose Agar medium was weighted as 3.9 gms and dissolved in 100 ml of distilled water and add 1gm of agar. Then the medium is kept for sterilization. After sterilization the medium was poured in a sterile petriplate, after solidification the Triplicates plates were aseptically spread on fungal suspension culture on the solid plates. The discs were kept over the agar plate using sterile forceps at various concentrations (50, 100 and 150 µl). These plates were incubated for 72 hrs at 37°C. After the incubation the plates were observed for formation of clear incubation zone around the disc indicated the presence of antifungal activity. The zone of inhibition was calculated.

 

RESULT:

In the present investigation, ethonolic leaf extracts of F. racemosa were subjected to preliminary phytochemical analysis. Preliminary phytochemical screening of the plant samples revealed the presence of tannins, Saponin, flavonied, terpenoied, steroids, glycosides, reducing sugar, resins, etc and the data is tabulated in (Table: 1). The extracts of plant sample revealed the presence of different phytochemical based on their polarity, extracting those plant metabolites of hydrophilic and hydrophobic nature and that the absence of anthocyonins in F. racemosawas significant.

 

Table:1 Phytochemical analysis of ethonolic extract of F.racemosa

S.

No

Phytochemical Constituents

F.racemosa plant extract

1.

Tannins

+

2.

Saponin

+

3.

Flavonoids

+

4.

Steroids

+

5.

Glycosides

+

6.

Terpenoids

+

7.

Resins

+

8.

Anthocyonins

-

 

The antibacterial activities of the ethonolic leaf extract of F.racemosa showed significant variations as shown in (Table: 2). the plant extract showed antibacterial activity against the pathogenic bacteria of Escherichia coli, Pseudomonas and staphylococcus.  The ethonolic leaf extract of F. racemosa was moderate activity against the (Escherichia coli, and staphylococcus at all a concentration and found non active against Pseudomonas at all concentration.  The plant extract presented moderate antibacterial activity and showed the very low antibacterial activity was recorded in (Table: 2).

 

Table:2  Antibacterial activity of the ethanol extract of F. racemosa

Test organisms

Zone of inhibition (mm) for different concentration

50µl

100µl

Control

Escherichia coli.

6

10

12

Pseudomonas

0

0

0

Staphylococcus

10

14

8

 

 

                      E.coli                                  Pseudomonas

 

Staphylococcus

Fig: 1 Antibacterial activity of the F.racemosa plant extract

 

Evaluation of the test plant extract for antifungal activity (Table:3) indicated that the ethonolic leaf extract were moderately active against Rhizopus at all concentration and found non active against A. Flavors and Trichodermaboth at 100µl concentration. The plant extracts presented moderate antifungal activity and showed the very low antifungal activity was recorded same table. Antimicrobial activity demonstrated that the activity of extract against bacteria, and fungi was mentioned of the board spectrum of activity. It can be used to source of the antibiotic substance for drug development and also used in the control of these bacteria and fungus infection.14

 

                      A.flavus                                  Rhizopus

 

Trichoderma

Fig:2 Antifungal activity of Ficus racemosa plant extract

 

Table:3 Antifungal activity of the ethanol extract of F. racemosa

Fungi name

Zone of inhibition (mm) for different concentration

50µl

100µl

Control

Aspergillus. flavus

3

0

0

Rhizopus

6

4

4

Trichoderma

4

0

0

 

DISCUSSION:

Medicinal plants are the most easily available productive sources of new compounds and drugs of natural origin. Phytochemicals are naturally occurring: biologically active and non- nutritive chemical compounds present in plants. Different type of phytochemicals have been known to possess medicinal properties and used in Indian system of traditional medicine.15 The phytochemical analysis result indicated the presence of flavonoied, glycoside, steroids and tannins as the main constituents that might be responsible for antimicrobial activity of the plant extract. The results indicate that the F.racemosa plant hold promises as sources of pharmaceuticals important phytochemicals. They are also involved in protective function in animal and have been usedas medicine.16

 

Generally an antibacterial and antifungal activity indicated that the inhibitory effect of the extracts depended on the type of plant species used, method of extraction and time of application of the extract.17 Evaluate the antimicrobial performance of an ethonolic leaf extract of F. racemosa against three different pathogenic bacteria and three different fungi isolates. All concentration showed to inhibit oral plant extract with a zone of inhibition about 6-14mm in antibacterial activity. By increasing the dose of the plant extract minimum inhibitory zone was found.There are several reports in the literature indicating the antibacterial and antifungal activity of the medicinal plants. Many studies reported the incapability of herbal antimicrobial agents to inhibit growth of gram –negative bacteria. Due to the presence of complex cell wall structure which decrease the penetration of bacterial cells by herbal extract.18but in the present study of F. racemosa leaf extract moderately inhibited the growth of bacteria and fungi proving penetrating ability to extract into microbial cells.

 

CONCLUSION:

The present study concludes that F. racemosa is an important medicinal plant with variable pharmacological spectrum. The phytochemical analysis revealed biologically active phytochemicals were present in the ethonolic leaf extract of F.racemosa. The result showed that the ethanol extract of F. racemosa was able to inhibit all of the bacteria and fungi used in this study with different degree of inhibition. This medicinal plant extract of F. racemosa is more effective against the tested bacteria than the fungi.

ACKNOWLEDGEMENT:

The authors are thankful for, the Management head, Department of Biotechnology, VELS University Pallavaram, Chennai, for giving the facility of doing the research work.

 

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Received on 05.12.2016             Modified on 06.01.2017

Accepted on 18.01.2017           © RJPT All right reserved

Research J. Pharm. and Tech. 2017; 10(2): 537-540.

DOI: 10.5958/0974-360X.2017.00107.X