Mouthwash from Banana Peel of Kepok Kuning Waste (Musa acuminate balbisiana

colla) as a Prevention of Streptococcus mutans Bacteria Growth

Causes of Dental Carries

Ai Rian Julyanti, Nenden Setyaningrum, Rosi Eka Ferliati,

Okta Nurandi Pratama, Erza Genatrika

Faculty of Pharmacy, University of Muhammadiyah Purwokerto,

Jl. Raya Dukuhwaluh, PO. BOX 202 Purwokerto 53182, Indonesia
(E-mail: airianjulyanti07@gmail.com)

ABSTRACT

Banana peel of kapok kuning (Musa acuminate balbisiana Colla) contains alkaloids,
flavonoids and tannins. These compounds are known to have antibacterial activity. This
study aims to determine the antibacterial activity of mouthwash from banana peel of
kapok kuning waste in inhibiting the growth of Streptococcus mutans. The process of
extraction of banana peel of kapok kuning using maceration method with 96% ethanol
solvent, then made formulation of mouthwash preparation with addition of extract
concentration 2, 4, and 8%. The antibacterial activity of the preparations was then
tested against Streptococcus mutans using agar diffusion method. The results showed
that the addition of banana peel of kapok kuning extract of 2, 4, and 8% in the mouth
rinse formulations can affect the inhibitory zone diameter of 9.52, 10.67, and 12.36 mm,
respectively. Mouthwash with the addition of 8% extract concentration has the greatest
antibacterial activity with a inhibition zone diameter of 12.36 mm. The extract inhibition
zone results and the formula yields p<0.05 i.e each test group has a difference
significant.

Keywords: Mouthwash, banana peel of kapok kuning, maceration, antimicrobial,

Streptococcus mutans, agar diffusion.

INTRODUCTION
Teeth are a valuable asset for humans which is a mirror of human health itself. Many
diseases that will arise starting from the lack of maintaining healthy teeth. Putri et al.
(2010) says that dental plaque is one of the problems in oral and dental health which is a
soft deposit attached tightly to the teeth, consisting of microorganisms that breed if
someone neglects tooth hygiene and mouth. In dental plaque there are various bacteria
and metabolism results. As an example of carbohydrate metabolism results by
asidogenic bacteria will result in the formation and accumulation of acid which resulted
in the decalcification and destruction of the tooth surface resulting in caries. Colonies of

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bacteria found in early plaque formation are Streptococcus mutans bacteria that many
experts believe to be the main cause of caries in teeth (Pratiwi, 2008).
Basic Health Research (Riskesdas) in 2013 states that the prevalence of carries through
the examination of Decayed Missing Filled Teeth (DMF-T) for the average shows 25.9%
of Indonesian people have dental and oral health problems. Prevention can be done by
inhibiting the growth of bacteria that cause infections using a variety of ways, one of
which is to rinse using mouthwash. Mouthwash is used because of its ability to
effectively reach a place that is difficult to clean with a toothbrush and can prevent the
formation of plaque (Lulun, 2012).

Based on data of banana production (Musa paradisiacal) from the Directorate General
of Horticulture Ministry of Agriculture can be estimated that the amount of banana peel
waste (Musa paradisiaca L.) in 2014 amounted to 2,745,023.2 tons. Currently the
researchers do a lot of research on medicinal plants as an alternative to chemicals that
already exist. One of the herbs that can be used as a medicine and has antibacterial
activity is banana peel of kapok kuning from the tribe of Musaceae. Previous studies
have shown that in banana peels there are flavonoids, alkaloids, saponins, tannins
according to Ogbonna et al. (2016). In alkaloid compounds, flavonoids, saponins, and
tannins are able to inhibit bacterial growth.

Based on these reasons, this research was conducted to make a mouthwase formulation
which is efficacious as antibacterial which is seen by comparing the obstacle zone
between ethanol extract and formulation of mouthwash preparation with the addition
of banana peel of kapok kuning extract in inhibiting the growth of Streptococcus mutans
bacteria. This research is expected to get the products of natural mouthwashes that are
effective, safe, affordable and can be produced on an industrial scale.

EXPERIMENTAL

2.1 Materials
The tools used in this study are rotary evaporator (IKA®10 Bassic), analytical scale
(Shimadzu), a set of extraction tools (Pyrex), drying cabinet, micropipet (Eppendorf),
yellow and blue tip, waterbath (HWB-3F -27L), pH sticks (Merck), viscometer
(Brookfield), UV-visible spectrophotometer (Shimadzu), and various types of

Proceeding of The 2nd UMP-PIC & 8th ISCC

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 glassware, autoclave (All American), incubator (Memmert), slip term, rounded ose,
bunsen, oven (Memmert), hot plate (IKA®C-MAG HS 7), centrifugator (Boeco),
tweezers, refrigerator (Sanyo). Materials used in this study were culture of
Streptococcus mutans, banana peel of kapok kuning, Mg powder (Merck),
concentrated HCl (Merck), FeCl3 1%, HCl 2N (Merck), dragendrof reagents, mayer
and wagner reagents (Merck), dimethylsulfoksida (DMSO), ethanol 96%
(Bratachem), TSA medium (Tryptone Soya Agar), TSB (Trypticase Soy Broth), sorbitol
(Bratachem), menthol (Bratachem), Na. saccharin (Bratachem), glycerol
(Bratachem), aquadest (Bratachem), gargle preparations in the market, filter paper,
and aluminum foil.

2.2 Methods
This research is a pure experimental laboratory (true experimental) study, with a
post-test design of complete randomized design in vitro. There were 3 groups of
yellow banana peel concentrations, 2, 4, and 8%, and negative control group,
aquades, povidone iodine mouthwash as positive control. Antibacterial power
testing is done by Disc-Paper Diffusion Method.
2.3 Extraction of banana peel of kepok kuning waste
Extraction of banana peel of kapok kuning waste is done by using maceration
method. Samples (500 g) of powdered banana peel of kapok kuning waste with a
solid solvent ratio of 1:2 ethanol 96% as a solvent 2-3 days until all compounds are
perfectly attracted, with several stirring then filtered and filtered to produce banana
peel of kapok kuning extract. The liquid extract was concentrated using a rotary
evaporator and waterbath at 40 °C until a thickened extract (Annie, 2012) was
obtained.
2.4 Phytochemical screening
Phytochemical screening performed on thick extract in the form of banana peel of
kapok kuning waste obtained in accordance with the method described by Rijayanti
(2014).
2.5 Mouthwash formulation banana peel of kepok kuning waste
The composition of the amount of variation of mouthwash formulation can be
explained in Table 1.
2.6 Evaluation of mouthwash banana peel of kepok kuning waste

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 Formula mouthwash preparations are physically evaluated including organoleptics
(color, odor, taste, turbidity, sediment), viscosity, acidity (pH), antibacterial power.
Viscosity was measured using Brookfield (DV2T) with spindle no. 61 and played at a
speed of 30 rpm.
Table 1. Formulation of banana peel of kapok kuning waste
Material FI (2%) F II (4%) F III (8%) K (-)
Banana peel of kapok kuning extract 2 4 8 -
Glycerine 15 15 15 15
Propyleneglycol 10 10 10 10
Na sakarin 0,1 0,1 0,1 0,1
Menthol 0,25 0,25 0,25 0,25
Distilled water (ad) 100 ml 100 ml 100 ml 100 ml
Source: Anastasia et al, 2017

2.7 Test of antibacterial extract and formula of mouthwash banana peel of kapok kuning
waste
Banana Peel an antibacterial power test was performed by Disc-Paper Diffusion
Method (Jawetz et al., 2005). The results of the antibacterial assay power were
based on the measurement of Area Diameter Hampers bacterial growth formed
around the paper disc (Ningsih, 2013).

RESULTS AND DISCUSSION

The results of phytochemical screening of banana peel of kapok kuning waste be seen in
Table 2. Observations of physical properties are recorded in Table 3.

Table 2. Phytochemical results of banana peel of kepokkuning waste

No. Test Banana peel of kapok kuning
1. Alkaloids +
2. Flavonoids +
3. Tannins +
4. Saponins No test
Keterangan: (+) positif (-) negative

Table 3. Result of evaluation of physical properties

No. Parameter Extract 2% Extract 4% Extract 8% Negative
Control
F1 F2 F3 F4
1. pH 5 5 5 6
2. Color Pale yellow, Yellow Dark yellow Bening

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 3. Odor typical menthol typical menthol typical menthol typical menthol
and the banana and the smell of and the smell of
smell is weak banana was banana is strong
4. Tasste Sweet and warm Sweet and warm Sweet and warm Sweet and warm
5. Turbidity Turbid rather Turbid rather Turbid rather Turbid rather
6. Sediment No No No No
7. Viscosity test 2 cP 2.2 cP 2.4 cP 2.2 cP
8. Antibacterial test 9.52 mm 10.67 mm 12.36 mm 0 mm

Fig 1. The zone of inhibitor mouthwash of

banana peel of kapok kuning waste.

Table 4. The result of measurement of diameter of mouthwash barrier with the addition
of banana peel of kapok kuning extract (Musa acuminate balbisiana Colla) to
the growth of Steptococcus mutans
Replication
The concentration of The average diameter
No
the dosage mouthwash of the inhibition (mm)
I II III
1. Formulation I (2%) 11.67 9.17 7.72 9.52
2. Formulation II (4%) 10.17 10.67 11.17 10.67
3. Formulation III (8%) 12.67 14.24 10.17 12.36
4 Control (+) 21.9 21.4 24.9 22.73
5. Control (-) - - - -

Based on the average measurement of the resistance diameter around the disc paper on
the mouthwash preparation added banana peel of kapok kuning extract at 24 hours
incubation period i.e the formulation showing the largest diameter is the formulation of
III with the concentration of 8% with the average inhibitory zone diameter 12.36 mm,
then formulation II with a concentration of 4% with an average resistor zone diameter of
10.67 mm and formulation I with a 2% concentration with a mean inhibitory zone
diameter of 9.52 mm.

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In addition, also used a negative control of the preparations without the addition of
banana peel of kapok kuning extract that does not show any inhibitory power. This
proves that the additional ingredients used do not affect the growth of bacteria in
mouthwash preparations. The outstanding supply in the market as a positive control
produces an average resistance power diameter at a 24-hour incubation period of 22.73
mm.

The purpose of the use of a positive control to compare the diameter of inhibitory
power supply in the market with the resulting mouthwash. Based on research Jannata et
al. (2014), the measurement of antibiotic strength of bacteria based on the David-Stout
method, states when the clear zone diameter ≤ 5 mm indicates weak antibacterial
activity, 5-10 mm diameter indicates moderate antibacterial activity, 10-20 mm
diameter indicates strong antibacterial activity, and > 20 diameter mm showed very
strong antibacterial activity. Based on this standard, the activity of banana peel of kapok
kuning extracton Streptococcus mutans is included in medium category. The larger the
diameter of the clear zone, the greater the antibacterial power. These antibacterial
compounds will result in greater osmotic pressure inside the cell, causing lysis. The
extract inhibition zone results and the formula yields p < 0.05 ie each test group has a
significant difference.

Phytochemical screening was performed to confirm the presence of secondary

metabolites, the results can be seen in Table 2. Extracts of banana peel of kapok kuning
positive alkaloids, flavonoids, and tannins. The presence of alkaloids is characterized by
the presence of yellowish white deposits at the base of the test tube, flavonoids are
marked with orange red to purple, and their tannins show a dark blue color (Rijayanti,
2014). From previous research conducted by Ehiowemwenguan (2014) also mentions
that banana peel contains alkaloid compounds, flavonoids, tannins, and saponins.
Alkaloids, flavonoids, and tannins include compounds that have antibacterial activity.

Antibacterial activity of banana peel kapok kuning extract was determined by

antibacterial test using agar diffusion method. Can be seen on Table 4, the result of
power barrier measurement is in medium strength range, where the inhibitory diameter
is in the range of 5-10 mm diameter. Inhibitory zone diameter ≤ 5 mm showed weak

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antibacterial activity and 10-20 mm diameter showed strong antibacterial activity, and >
20 mm diameter showed very strong antibacterial activity (Jannata et al., 2014).

The evaluation of the preparation includes organoleptic observation (color, odor, taste,
turbidity, sediment), viscosity test, acidity test (pH), antibacterial power. Viscosity was
measured using Brookfield (DV2T) with spindle no. 61 and played at a speed of 30 rpm.
The test is performed for 21 days of storage. The results can be seen in Table 3 where
organoleptically for 21 days the storage dosage does not show any change in color,
odor, and taste, turbidity, and sediment. Based on the result of viscosity test according
to parameter of cP water value that is 1 cP. The higher the concentration of banana peel
skin extract, the higher the viscosity and the pH value of the mouthwash according to
the parameters of the topical dose preparation for mouthwashes mentioned by Sakinah
(2016) that is between 5-6. If the pH < of 5 dosage is too acid and will cause more
bacteria growth and if pH > of 6 then the dosage is too alkaline and will cause the
growth of fungus, thus causing sprue.

CONCLUSIONS
Based on the results obtained, it can be concluded that the banana peel of kepokkuning
extract (Musa acuminate balbisiana Colla) can inhibit the growth of Streptococcus
mutans bacteria and is bacteriostatic. The greater the addition of extract concentration
to the mouthwash formula, the greater the inhibition zone will be produced.

ACKNOWLEDGMENT

This acknowledgment is aimed to the Director General of DIKTI Republic of Indonesia

which has been given an opportunity and research grant at Student Creativity Week-
Eksakta Research (PKM-PE) 2017 and also appreciation to Faculty of Pharmacy
University of Muhammadiyah Purwokerto which has been given facilities.

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Ehiowemwenguan., G., Emoghene, A.O., Inetianbor, J.E. Antibacterial and phytochemical
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