An In vitro Study on Bioactivity of Ficus racemosa

 

Antony V Samrot*, Raji P, Durga Sruthi P, Karishma S, Jenifer Selvarani  A, Nishanthini P

Department of Biotechnology, Sathyabama University, Jeppiar Nagar, Chennai, Tamil Nadu – 600 119, India

 

ABSTRACT:

In vitro evaluation of bioactive compounds is the simplest and effective method for evaluating the antimicrobial and antioxidant activities of compound. In this study, the leaves of Ficus racemosa were extracted with chloroform. The qualitative phytochemical screening was done for the extracted sample. The extract was checked for its antioxidant and antibacterial activity using the TLC bioautography assay. The well diffusion method was carried out to study about the antibacterial activity of the plant sample against three different bacterial cultures. The chloroform extract of Ficus racemosa was showing least antioxidant and antibacterial activity.

 

 

 

INTRODUCTION:

A large number of deadly diseases have emerged over the last few years and most sources to treat them are derived from plants¹. Plant derived materials are with minimal toxicity compared to chemically synthesized antibiotics. These phytoconstituents perform various biologically important temperaments like Antibacterial, Antioxidant, Antifungal properties etc^2–3. The traditional uses of Ficus racemosa include medicines to treat disorders like boils, diabetes, diarrhea, leprosy and measles etc^5,6. In this study, leaves of Ficus racemosa were collected from local area and chloroform extract was taken. The collected extract was subjected for in vitro evaluation for antibacterial and antioxidant activity.

 

MATERIAL AND METHODS:

Collection of plant and preparation of extract

The leaves of Ficus racemosa were collected from Sathyabama University campus, Jeppiaar Nagar, Chennai, Tamil Nadu (Latitude: 12°52' 13.79" N and Longitude: 80°13' 11.40" E). The collected leaves were washed several times under tap water. The leaves were then shade dried for 2 weeks and later ground into fine powder. The powdered leaves were used for further extraction. To prepare the extract, 10g of the powdered plant sample was measured and added to 100ml of chloroform (1:10 ratio) and the mixture was kept in a shaker incubator overnight. The solution was then filtered using Whatmann filter paper No.1. The filtrate obtained was concentrated by keeping it in a rotary evaporator^{4, 7}.

 

PHYTOCHEMICAL ANALYSIS:

To find out the various metabolites that might be present in the plant extract, various chemical tests were performed ^{8, 9}.

 

SEPARATION OF BIOACTIVE COMPOUNDS BY THIN LAYER CHROMATOGRAPHY:

The chloroform extract of Ficus racemosa was subjected to Thin Layer Chromatography by using TLC silica plate (Merck, F245). Five different mobile phases namely Chloroform, Methanol, Dichloromethane, Ethanol and a combination of Chloroform and Methanol were used. The Retention factor (Rf) was noted from the bands formed in the TLC plate after exposure to iodine.

 

TLC BIO-AUTOGRAPHY FOR ANTIOXIDANT ACTIVITY:

The extracts were chromatographed using TLC silica plate, plates were dried, sprayed with DPPH (0.004% w/v in 95% methanol)^4,10,11. The Rf value was determined.

 

TLC BIO-AUTOGRAPHY FOR ANTIBACTERIAL ACTIVITY:

TLC plates ran with the extracts were air dried, sprayed with 24h culture of Pseudomonas aeruginosa and incubated at 37̊ C for 24h. After incubation it was sprayed with MTT reagent (0.001%w/v in Distilled water) and was observed for antibacterial activity^{4, 12}. The Rf values were recorded.

 

ANTIBACTERIAL ACTIVITY BY AGAR WELL DIFFUSION METHOD:

The antibacterial activity of the plant extract was tested against 3 organisms namely Pseudomonas aeruginosa, Bacillus subtilis, Brevibacillus brevis by performing the agar well diffusion assay.

 

RESULTS AND DISCUSSIONS:

The results for the phytochemical screening with the chloroform extract of Ficus racemosa is represented in the (Table 1). It was identified with maximum presence of metabolites like carbohydrates, glycosides and diterpenes. An adequate quantity of saponins, tannins and traces of amino acids were also spotted. The solvent extract lacked the presence of other metabolites like alkaloids, phytosterols, phenols and flavonoids. The leaves of this plant are reported to have flavonoids, triterpenoids, alkaloids, and tannins¹³. Tannins and cardiac glycosides were also reported in ethanol and methanol extracts of F.racemosa¹⁴.

 

Most components have been separated when chloroform was used as mobile phase (Fig.1). The Rf value of the components are listed in (Table 2). Most phytochemicals were found to be separated in ethyl acetate and acetone extracts and the Rf value were also elucidated¹⁴. Antioxidant active components were found to be separated when methanol and ethanol were used as mobile phase (Fig.2, Table 3). The mobile phase might have separated the components because of their polarity. Ficus racemosa has been reported to have antioxidant and radioprotective activity¹⁵. When the extract was subjected for TLC bioautography for antibacterial activity against Pseudomonas aeruginosa, there was no activity (Fig.3). It was confirmed with antibacterial activity using agar well diffusion method (Fig.4), but it was showing antibacterial activity against Brevibacillus brevis (Table 4). Mandal et al (2000) found the leaves extract of F. racemosa to have antibacterial activity against Gram positive and Gram negative bacteria.

 

Table 1: Phytochemical analysis with the Chloroform extract of Ficus racemosa

+++   indicates Immediate change

++     indicates change which occurred within 5 min

+       indicates change that occurs after 5 min

-        indicates no such change

 

Fig.1 TLC ran with different mobile phase and exposed to iodine a) chloroform b) dichloromethane c) ethanol d) methanol e) chloroform and methanol

Table 2: Rf value of TLC ran with different mobile phase and exposed to iodine

 

Fig.2 TLC Bioautography for antioxidant activity for TLC ran with different mobile phase a) chloroform b) dichloromethane c) ethanol d) methanol e) chloroform and methanol

 

Table 3: Rf value of TLC bioautography for antioxidant activity for TLC ran with different mobile phase 

 

Fig.3 TLC bioautography for antibacterial activity against Pseudomonas aeruginosa for TLC ran with different mobile phase  a) chloroform b) dichloromethane c) ethanol d) methanol e) chloroform and methanol

 

Fig.4 Antibacterial activity of Ficus racemosa a)Pseudomonas aeruginosa b) Brevibacillus brevis c) Bacillus subtilis

Table.4: Antibacterial activity of Ficus racemosa a)Pseudomonas aeruginosa b) Brevibacillus brevis c) Bacillus subtilis

 

REFERENCE:

1.     Rout SP, Choudary KA, Kar DM, et al. Plants in traditional medicinal system—future source of new drugs. International Journal of Pharmacy and Pharmaceutical Sciences. 2009; 1(1).

2.     Sashikala GD, Kottai AM, Satheesh DK, Rekha S, Indhumathy, et al. Studies on the antibacterial and antifungal activities of the ethanolic extracts of Luffacy lindrica (Linn) fruit. Int J Drug Dev. Res 1: 2009: 105-109.

3.     Orak HH, Demirci A, Gümü T. Antibacterial and antifungal activity of Pomegranate (Punica granatum L. CV.) Peel. EJEAFCh. 10: 2011: 1958-1969.

4.     Samrot AV, Bennet Rohan, Divya Kumar, Sahiti K, P. Raji, Sree Samanvitha K. Detection of antioxidant and antibacterial activity of Mangifera indica using TLC bio-autography. IJPSR. 7(11): 2016: 4467-4472.

5.     Siwakoti M, Siwakoti S. Ethnomedicinal uses of plants among the Satar tribe of Nepal. In: Maheswari, J.K. (Eds.), Ethnobotany and Medicinal Plants of Indian Subcontinent. Scientific publisher. 2000: 98-108

6.     Paarakh PM. Ficus racemosa Linn: an overview. Natural Product Radiance. 8 (1):2009:84–90.

7.     Suleiman MM, Mcgaw LJ, Naidoo V, Eloff JN: Evaluation of several tree species for activity against the animal fungal pathogen Aspergillus fumigatus South African Journal of Botany. 76: 2010: 64–71

8.     Jamal AK, Yaacob WA, Laily BD, A Chemical Study on Phyllanthus reticulatus. J Physical Sci. 19 (2): 2008: 45–50

9.     Haque MA, Hassan MM, Das A, Begum B, Ali MY and Morshed H, Phytochemical investigation of Vernoniacinerea (Family: Asteraceae), J. Appl. Pharma. Sci. 02(06): 2012: 79-83.

10.   Cimpoiu DCJ. Analysis of some natural antioxidants by thin-layer chromatography and high performance thin layer chromatography. J Liq Chromatogr R T. 7–8: 2006:1125– 1142.

11.   Takao T, Kitatani F, Watanabe N, Yagi A, Kanzo Sakata: A Simple Screening Method for Antioxidants and Isolation of Several Antioxidants Produced by Marine Bacteria from Fish and Shellfish. Biosci. Biotechnol. Biochem. 58: 1994: 1780–1783.

12.   Silva MTG, Simas SM, Batista TG, Cardarelli P, Tomassini TC: Studies on antimicrobial activity, in vitro, of Physalis angulata L. (Solanaceae) fraction and physalin B bringing out the importance of assay determination. Mem. Instit. Oswaldo Cruz. 100(7): 2006: 779–782.

13.   Shiksharthi AR, Mittal S. Ficus racemosa: Phytochemistry, traditional uses, and pharmacological properties: A review. Int J Adv Pharm Res. 4: 2011: 6-15.

14.   Ganatra SH, Durge SP, Patil SU. Preliminary Phytochemicals Investigation and TLC Analysis of Ficus racemosa Leaves. Journal of Chemical and Pharmaceutical Research. 4(5): 2012: 2380-2384.

15.   Veerapur VP, Prabhakar KR, et al. Ficus racemosa Stem Bark Extract: A Potent Antioxidant and a Probable Natural Radioprotector. NCBI. v. 6(3); 2009.

16.   Mandal SC, Saha BP, Pal M. Studies on antibacterial activity of Ficus racemosa Linn. leaf extract. Phytother Res. 4(4): 2000: 278-80.

 

 

 

 

Accepted on 11.09.2017        © RJPT All right reserved

Research J. Pharm. and Tech 2017; 10(12): 4219-4222.