The comparative study of the fruit and leaf extract of Ficuslyrata Warb on antibacterial activities

Abstract

The extraction process of anti-microbial compounds from F.lyrataWarb fruit and leaf had been identified. The study employed a maceration extraction method used water as solvent agents, to extract seconder metabolite, example phenolic, flavonoid, tannin as an anti-microbial agent. The compound which is known to have an inhibition effect to decline the growth of the microorganism of food quality decreasing triggers. Its effects were subjected to 3 tested bacterias, i.e.; Pseudomonasaeruginosa, Escherichia coli, and Bacillus subtilis. This study purpose to determine and compare the content of phytochemicals and antimicrobial activity in leaf and fruit extracts. Experimental laboratorium was conducted and the data analyzed by descriptive analysis in this research. The statistical test used is two sample tests, which aims to know the difference in mean value between leaf extractwith fruit, which is not mutual related. Results indicated the formation of the inhibited zone in the tested media which proved by the Kirby-Bauer testing method are different between fruit and leaf extract.

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The comparative study of the fruit and leaf extract of
Ficuslyrata
Warb on
antibacterial activities
To cite this article: Dwi Wahyudha Wira et al 2018 IOP Conf. Ser.: Mater. Sci. Eng. 420 012077
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2nd Nommensen International Conference on Technology and Engineering IOP Publishing
IOP Conf. Series: Materials Science and Engineering 420 (2018) 012077 doi:10.1088/1757-899X/420/1/012077
The comparative study of the fruit and leaf extract of FicuslyrataWarb on
antibacterial activities
Dwi Wahyudha Wira1, Efri Mardawati2, Endang Yuni Setyowati1, Anisa
Dahlan3, Roostita L, Balia1
1Study Program of Veterinary Medicine, Faculty of Medicine, Universitas
Padjadjaran, Jatinangor Km21, 40600
2Dept. of Agroindustrial Technology, Universitas Padjadjaran, Jatinangor Km21
3Study Program of Medicine, Faculty of Medicine, Universitas Padjadjaran,
Jatinangor Km21
Email: dwi.wahyudha@unpad.ac.id
Abstract. The extraction process of anti-microbial compounds from F.lyrataWarb
fruit and leaf had been identified. The study employed a maceration extraction
method used water as solvent agents, to extract seconder metabolite, example
phenolic, flavonoid, tannin as an anti-microbial agent. The compound which is
known to have an inhibition effect to decline the growth of the microorganism of
food quality decreasing triggers. Its effects were subjected to 3 tested bacterias,
i.e.; Pseudomonasaeruginosa, Escherichia coli, and Bacillus subtilis. This study
purpose to determine and compare the content of phytochemicals and
antimicrobial activity in leaf and fruit extracts. Experimental laboratorium was
conducted and the data analyzed by descriptive analysis in this research. The
statistical test used is two sample tests, which aims to know the difference in
mean value between leaf extractwith fruit, which is not mutual related. Results
indicated the formation of the inhibited zone in the tested media which proved by
the Kirby-Bauer testing method are different between fruit and leaf extract.
1. Introduction
One of the natural medicinal plants have been used for antimicrobial resources is a Ficus
plant. In Indonesia, the most important species of Ficus are F. lyrataWarb, F. elastica, and
Ficuscarica. Various parts of the plant like bark, leaves, stem, fruits, seeds, and fruit are
medicinally important [1]. Ficus usually grows in subtropical and tropical climate areas. Fruits
and leaf of ficus trees are widely used in the fresh or dried form. They are an excellent source
of minerals, vitamins, amino acids, crude fibers as well as phenolic compounds [1]. This
natural material extracted on the skin of the roots and stems to produce antimicrobial [2].
Compounds contained in the root bark and stems are flavonoids, alkaloids, tannins, phenolic
compounds and terpenoids compounds that can inhibit the growth of pathogenic bacteria. The
fruit extract of Ficussycomorous L, Ficusbenjamina, Ficusbengalensis L andFicusreligiosa
showed antibacterial and antitumor activity [3]. Due toextract of F. lyrataWarb fruits and leaf
could show antimicrobial.
The increased of usage the chemical preservative is dangerous to human health.
Literature reports and ethnobotanical records suggest that plants are potential of the
pharmaceutical industry. They may provide a natural source of antimicrobial agents that
willor provide novel or lead compounds that may be employed in controlling some infections

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2nd Nommensen International Conference on Technology and Engineering IOP Publishing
IOP Conf. Series: Materials Science and Engineering 420 (2018) 012077 doi:10.1088/1757-899X/420/1/012077
globally[1]. The fruit and leaf extract of F. lyrataWarb contain the bioactive compounds,
especially phenolic compounds, flavonoids, triterpenoids, and tannins [2]. The application of
natural preservatives in the food industry, especially in the preservation of meat can improve
food safety. The aim of this study is to compare the potential utilization of F. lyrata fruit and
leaf extract as antimicrobial agents. The predicted result of this research is a difference the
concentration of compound bioactive and antimicrobial activity between fruit and leaf
extract.
2. Material and Methods
2.1. Raw materials
Fresh fruits and leaves of F. lyrataWarb or plant parts were collected randomly from
Universitas Padjadjaran, Indonesia. Fresh fruits leaf materials were washed under running tap
water, air dried and then ground to fine powder and stored in airtight bottles.
2.2. Crude extraction
The Dried Powder of fruit and leaf of 200 g of was extracted in 1600 mL of distilled water
for 48 h at room temperature[1]. Extraction was conducted in sealed flasks, and then it was
filtered through vacuum filtered. The supernatant was collected. It was then rotavaped to
result in the concentration of extract up to 50 % at 40 ºC. All experiments were performed in
duplicate to check the reproducibility.
2.3 Phytochemical screening
The extracts were screened for phytochemicals like alkaloids, flavonoids, triterpenoids,
phenolic compounds,and tannins, following the procedure of Harborne[4].
Test for phenols (Ferric chloride test) [4], 2 mL of sample was treated with aqueous ferric
chloride 5% and observe for the formation of deep blue or black color. Test for tannins
(Ferric chloride test) [5], 2 mL of sample was treated with aqueous ferric chloride 1% and
observe for the formation of deep blue or black color. Test for flavonoids (NaOH 10% test)
[5], 1 mL of sample was treated with NaOH 10% and observed for formation of brownish
orange. Test for triterpenoids (Lieberman-Burchard test) [6], 1 mL ofthe sample was
treated with 2 mLof acetic anhydride and 2 mL of H2SO4. The formation of dark red indicates
the presence of triterpenoids. Test for alkaloids (Mayer’s reagent) [5], 2 mL ofthe sample
was treated with 2 mL of Mayer’s reagent. The formation of dark red indicates the presence of
triterpenoids. Test for saponins (Foam’s test) [4], 5 mL of sample in the test tube was heated
for 1 minute in 55-60oC. The test tube was shaken vertically for around 5 minutes. The foam
that formed indicated the presence of saponins.
Total phenols (Folin-Ciocalteau method) [7], about 3 g sample was weighted and put
inside 50 mL volumetric flask. 0.5 ml sample was put inside a 25 mL volumetric flask then
was added 0.5 ml FolinCiocalteau and 10 mL of Na2CO3 7.5%. The absorbance was
measured in 750 nm with gallic acid as standard. Total Tannins (Folin-Denis method) [8],
About 3 g sample was weighted and put inside 50 mL volumetric flask. 0.5 ml sample was
put inside a 25 mL volumetric flask then was added 1.25 mL ofFolin Denis and 2.5 ml
Na2CO3. The absorbance was measured in 760 nm with tannic acid (0, 2,0 ; 4,0 ; 6,0 ; 8,0 ;
10,0 ppm) as standard. Total Flavonoids[9], abouts 3 g sample was weighted and put inside
50 mL of volumetric flask. 0.5 ml sample was put inside 25 ml volumetric flask then was
added 0.30 mL of NaNO2 5%, 0.30 mL of AlCl3 10%, and 2 mL of NaOH 1M. The

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2nd Nommensen International Conference on Technology and Engineering IOP Publishing
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absorbance was measured in 510 nm with quercetin (0, 2,0 ; 4,0 ; 6,0 ; 8,0 ; 10,0 ppm) as
standard.
2.4. Analysis antibacterial activity
Antibacterial activity for the fruit and leaf extracts was evaluated by disc diffusion method
Kirby Bauer[10]. The bacteria used in the study include Pseudomonasaeruginosa, Escherichia
coli, and Bacillus subtiliswere obtained from the Pharmacy Faculty, Universitas Padjadjaran,
Indonesia. The bacterial isolates were first sub cultured in a nutrient broth and incubated at
37°C for 36 h. Amoxycillin (250 mg) was used as a standard antibiotic for comparison of the
results. Muller-Hinton agar medium was loaded into Petri dish, stored until freezing and then
inoculated tested bacterial to medium. The diameter of the zone of inhibition was measured
in millimeter (mm). The experiment was repeated in triplicates and the average values were
calculated. Phytochemical content data and antimicrobial activity were analyzed using two-
sample t-test to determine whether there was a difference between leaf and fruit extract[11].
3. Results and Discussion
3.1. Phytochemical Screening
Phytochemical compound screening of F. lyrata Warb fruit and leaf extracts was
conducted using a qualitative method including flavonoids, phenolic compounds,
triterpenoids and tannins from the dried fruit, but the triterpenoid tobe absent in leaf extract.
Phytochemicals such as alkaloids, triterpenoids, and saponin were found to be absent in both
extracts. The observed results the value of phytochemical screening were presented in table 1.
Table 1. Phytochemical screening the various extracts of FicuslyrataWarb
Phytochemical
compound
Result
Fruits
Leaf
Phenolic
+
+
Flavonoid
+
+
Alkaloid
-
-
Triterpenoid
+
-
Saponin
-
-
Tanin
+
+
Note : (+) : Present
(-) : Absent
Many of active compound like phenolic compounds in plants comprise a comparatively
large class of secondary metabolites with varies potential bioactivities, such as antioxidant,
antibacterial, and anti-inflammatory[12,13]. The amounts of total bioactive compounds in the
water extract was huge. It is generally known that water extract a better influence on the
extractability of phenolic compounds, moreover, water is more suitable for the food industry
than the other organic solvent. The comparative phytochemical compound of extracts of
FicuslyrateWarb fruit and leaf as shown in table 2.

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2nd Nommensen International Conference on Technology and Engineering IOP Publishing
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Table 2. Various Phytochemical compound of extracts of fruit and leaf Ficuslyrata Warb
Phytochemical
compound
Value (mg/g)
Value (mg/g)
Fruit
Leaf
Phenolic
0.38a±0.04
1.67b±0.21
Flavonoid
0.13a±0.01
1.22b±0.25
Tanin
0.39a±0.17
0.95b±0.13
Table 2 show that the bioactive compounds found in both types of the extract are phenol,
flavonoid,and tannin. The content of phenol, flavonoid and tannin compounds on leaves is
significantly different from the compounds contained in the fruit. Where the bioactive
component is higher. This suggests that the leaf extract is more potentially used as an
antimicrobial agent.This is similar with the research reported Rajiv[12], Ficusreligiosa fruit
extract gave positive results on phytochemical qualitative tests of phenolic compounds,
flavonoids, terpenoids,andglycosides. In addition, Adebayo[14], reported the results of
qualitative tests performed on ficus exasperate leaves gave positive results on tannin,
flavonoid, and saponin components.
3.2. Antibacterial activity
The antimicrobial activity test aims to explore the antimicrobial potential resulting from an
ingredient by looking at the clear zone formed. In order to see the clear zone formed by the
diffusion method[15]. This is based on antimicrobial activity can be seen from the clear zone
formed around it. The clear zone shows the resistance of microbial growth in the solid layer in
the petri dish.
In the evaluation of antibacterial activity of the fruit and leaf extract, zone of inhibition was
observed in the extracts of water. Microbial growth was determined by measuring the
diameter of the zone of inhibition. For each bacterial strain, controls were maintained in
which pure solvents were used instead of the extract. The antibacterial activity of aqueous
extracts against of 3 bacteria is shown in figure 1.
Figure 1. Inhibition zone of extracts against of the different bacteria and different extract
Maximum antibacterial activity was shown by Bacillus subtilis from leaf extract while
minimum activity was shown by P. Aeroginosa from leaf extract. From the results of
phytochemical screening analysis and antibacterial activity, it is clear that the presence of
phytochemicals compound including flavonoids, phenolic and tannins are essential material
Pseuodomonas
aeruginosa Eschericia coli Bacillus subtilis
2.29a3.98a4.72a
1.11a
4.33a
6.66a
Fruit Extract (mm) Leaf Extract (mm)

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for the inhibition of bacterial growth. Inhibition zone of B.subtilis and E. coli in leaf extract
is higher than in fruit extract. On the other hand, in P. Aeroginosa bacteria, the inhibitory
zone of the fruit is higher than leaf extract. This result indicates that the antimicrobial activity
of leaf extract is more potentially use as biopreservative to against the B.subtilis and E. Coli.
This is because the content of bioactivities such as phenolic, flavonoid,and tannin is higher in
leaf extract than in fruit. When compared with similar research conducted on ficus plants
such as Ficusexasperata extracted using methanol solvent yielding a clear zone diameter on P.
aeruginosa bacteria is between 3-6 mm[14]. This value is close to the result of research where
the clear zone of P. aeruginosa bacteria on ethyl acetate extract is 2.97 mm. Meanwhile
reported that the inhibitory zone of Ficusreligiosa plant using aquadest as the solvent is 5-9
mm with control is tetracycline with 13 mm inhibitory zone value[13].
Flavonoids and tannins are included in the phenol compounds that have an important role
in the biochemical process. Such compounds may cause major disruption because of their
ability to form protein complexes through hydrogen bonds. When the plant cell content mixes
with the membrane becomes damaged during the isolation process, the phenol compound
rapidly reacts to form a complex with proteins. As a result, there is often an enzyme inhibition
of plant extracts [4]. The mechanism of inhibition of phenol compounds that inhibit protein
synthesis and damage the action of enzymes[16]. This causes cell metabolism disorders and
destroys cell membranes by dissolving fat in cell walls. Factors affecting antimicrobial
activity are extract type, amount, a microbial source, antimicrobial agent concentration,
temperature, contact time, and physical properties of substrate chemistry (pH, moisture
content, surface tension, type and amount of solute, colloids,and compounds other
compounds). This mechanism occurs so that the F. Lyratafruit and leaf extractshave
antimicrobial activity.
4. Conclusion
This research showed that extracts of F. lyrataWarbfruit and leaf could use as
antimicrobial resources. The phytochemical screening resulted in the water fruit leaf extract
presence of secondary metabolites like flavonoids, phenolic, and tannin. Antimicrobial
activity was observed by the disc diffusion method against bacterial pathogens including
Pseudomonasaeruginosa, Escherichia coli, dan Bacillus subtilis. The result showed that the
leaf extract has the higher of bioactive compounds than fruit extract,on the other side is
antimicrobial activityis not a significant difference.
5. Acknowledgment
This research was funded by Universitas Padjadjaran Internal Grant of Unpad under the
scheme of Fundamental grant research 2018.
6. References
[1] Jagathambal, M, K.;Madhavan.; Parvathi.;Karunakaran,:Phytochemical Screening
Antimicrobial activity of Seven Solvent Extracts of Dried Fruit of Ficuscarica Against
Five Human Pathogenic Bacteria G.:Journal of Pharmacy Research, 4(12) (2011),
,4498-4500
[2] Bidarigh, S, P.; Khoshkholgh, A.; Massiha,;Issazadeh, K,: . Proceeding International
Conference on Biotechnology and Environment Management IPCBEE, vol 18 (2011)

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2nd Nommensen International Conference on Technology and Engineering IOP Publishing
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[3] Sirisha. N, M.; Sreenivasulu, K.; Sangeeta, C. M.; Chetty.: Antioxidant Properties of
Ficus Species-A Review, International Jurnal of pharmTech Research, 2(4), 2010.
[4] Harbone J B., “Phytochemical methods: A guide to modern techniques of plant
analysis”, 3rd Edn, Chapman and Hill, London, 1998
[5] Noor, H. Al Atraqchi., Wafaa. M. A., and Al Shaikh. H. 2014. Preliminary
Phyochemical Creening and In Vitro Evaluation of Antioxidant Activity of Iraqi
Species of SilybummarianumSeeds. International Research Jounral of Pharmacy, 2014 5
(5). ISSN 2230-8407
[6] Kokate, C.K., A.P. Purohit, dan S.B. Gokhale. 2008. Pharmacognosy. NiraliPrakashan,
Maharashtra
[7] Sagu, Sorel Tchewonpi, Sankha Karmakar, Emmanuel Jong Nso, and Sirshendu De.
2014. Primary Clarification of Banana Juice Extract by Centrifugation and
Microfiltration. Department of Chemical Engineering, Indian Institute of Technology,
Kharagpur, India
[8] AOAC. 2000. Official of Analysis. Association of Official Analytical Chemist. EUA
[9] Tambe, V. D., and R. S. Bhambar. 2014. Estimation of Total Phenol, Tannin, Alkaloid
and Flavonoids in Hibiscus tillaceusLinn. Woos Extracts. Journal of Pharmacognosy
and Phytochemistry Volume 2, Issue 4. ISSN : 2321-6182.
[10] Capuccino, J.G., N. Sherman., “Microbiology: A Laboratory Manual”, Sixth Edition,
Benjamin Cummings, San Fransisco, 2001
[11] Skaik, Y: The Bread and butter of Statistical Analysis “T-test: Uses and Misuses, Pak J
Med Sci, 2015 Nov-des; 31(6): 1558-1559
[12] Rajiv, P,; R. Sivaraj.: Screening For Phytochemicals And Antimicrobial Activity Of
Aqueous Extract Of Ficusregilosa Linn, Int. Jurnal of Pharmacology and
Pharmaceutical Sciences, 4 (5), (2012).
[13] Rizvi W., Rizvi M., Kumar R., Kumar R., Shukla I., Parveen M.: Antimicrobial
Activity of Ficuslyrata-An In Vitro Study, Int. J. Pharmacol, 8(2).7 (2010).
[14] Adebayo, E. A., O. R. Ishola, O. S. Taiwo, O. N. Majolagbe, B. T. Adekeye. 2009.
Evaluations of The Methanol Extract of Ficus exasperate Stem Bark, Leaf and Root fot
Phytochemical analysis and Antimicrobial Activities. African Journal of Plant Science
Vol. 3 (12).
[15] Jawetz, E., J. Melnick and E. Adelberg. 2004. Medical Microbiology 23rd ed. Mc-Graw
Hill Publ. Co. Ltd. New York.
[16] Davidson, P.M. (1993) Parabens and phenolic compounds. antimicrobials in Foods, 2nd
edn. ed. Davidson, P.M. and Branen,A.L. pp. 263–306. New York: Marcel Dekker Inc.