International Conference on Mathematics, Science, and Education 2016 (ICMSE 2016)

Hand Sanitizer with Pineapple Peel Extract as Antibacterial against Staphylococcus aureus and
Escherichia coli
1,a) 1 1
Nanik Wijayati , Anggy Rinela Sulistya Rini , and Supartono ,
1
Department of Chemistry, Faculty of Mathematics and Science, State Semarang University
a)
Corresponding author: nanikanang@gmail.com

ABSTRACT

Pineapple peel usually just thrown away as waste. According to some research in pineapple peel contains many active
subtances were effectively kills bacteria. Phytochemical test shows a positive result that pineapple peel extract contains
flavonoids, tannins and saponins. Analysis using UV-Vis spectrophotometer and FTIR showed that the extract
contains derivative of dihidroflavonon. In this research, pineapple peel extract is applied as hand sanitizer, then
tested the antibacterial activity against Escherichia coli and Staphylococcus aureus. Test results showed that hand
sanitizer at contrencation of 0,5%, 1% and 1,5% pineapple peel extract can kills bacteria. Optimum activity against
bacteria reached at 1,5% formulation, that is 15 mm for Escherichia coli and 15,5 mm for Staphylococcus aureus.
Moreover, all formulation meet the quality standards and pass the organoleptic test, pH, homogeneity and dispersive
power.

Keyword: pineapple peel, hand sanitizer, antibacterial

INTRODUCTION
Pineapple peel mostly just thrown away as waste,
but according to several studies pineapple peel
containing vitamin C, carotenoids and flavonoids.
Several studies have been conducted on the antibacterial
activity of the extracts of pineapple, among others Suerni
et al. (2013) examined the antibacterial activity of
extracts of pineapple fruit, salak and kweni mango
against Staphylococcus aureus is showed that pineapple
fruit extract can inhibit the growth of Staphylococcus
aureus at concentrations of 50% and 100%. Chanda et al.
(2010) have also studied the antibacterial activity of
extracts of pineapple peel with chloroform, acetone
and methanol, the results show chloroform extract of
pineapple peel have activity against Staphylococcus
aureus, Corynebacteriumubrum, Klebsiella pneumoniae
and S. typhimurium. Pineapple peel acetone extract
showed activity against Staphylococcus aureus, S.
subflava, Enterobacter aerogenes, Klebsiella
pneumoniae, Proteus mirabilis and S. typhimurium. The
methanol extracts showed activity against
Staphylococcus aureus and Klebsiella pneumonia.
From the studies that has been mentioned above,
prompting the authors to researching antibacterial
activity of pineapple peel and apply for hand sanitizer
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that aims to allow communities to better utilize
something that previously only be a waste.
METHODS
Tools that used for phytochemical test and
sample extraction is a knife, a watch glass, spatula,
analytical balance, test tubes, pipette, light spirits,
erlenmeyer, a set of maceration, rotary evaporator,
measuring cups, filter paper, and a stirrer. Tools that used
for the preparation of hand sanitizer is a mortar and
stamfer, stirrer, analytical balance, spatula, watch glass
and a pipette. The tools that used to test the inhibition of
bacteria is a petri dish, discs, ose, light
spirits/bunsen, erlenmeyer, autoclaves, incubators, flask,
graduated cylinder, calipers, stirrer, stirrer, label paper,
plastic wrap, tissue. While chemical instruments are used
to characterize the compounds in this study is the FTIR
and UV-Vis Spectrophotometer.
The materials used are pineapple peel as samples,
culture media Nutrient Agar (NA), Sodium
physiological, methylated spirits, H2SO4, FeCl3,
aquadest, n-hexane and ethanol concentrated. Test
bacteria used are Staphylococcus aureus and Escherichia
coli. While the materials needed to make gel hand
 International Conference on Mathematics, Science, and Education 2016 (ICMSE 2016)

Table 1. Hand sanitizer gel formulation of pineapple peel Table 2. Phytochemical screening results
extract

Information:
+ = contains the compound
• = not containing the compound

(Sari and Isadiartuti, 2006, with modifications)

sanitizer is a sample extract of pineapple peel, CMC,

TEA, glycerin, methyl paraben and distilled water.
Extraction is used maceration method with n-
hexane and continued with ethanol and macerated for 6
days. (Solikhah, 2014, with modifications). Pineapple
peel extract is used for phytochemical screening,
analyzed using UV-Vis and IR and active agent in hand
sanitizer.
Hand sanitizer gel formula draws on research by
Isadiartuti and Sari (2006) with modifications, where
betel leaf extract is an active ingredient in the formula of
hand sanitizer replaced with pineapple peel extract and
sodium metabisulfite is replaced with methyl paraben.
Preparation of a gel hand sanitizer using the method of
Shu (2013) with modifications. The hand sanitizer gel
formulations with the active ingredient pineapple peel
extract can be seen in Table 1.
Antibacterial activity test was conducted using
disc method and what is tested is extracts of pineapple
peel, hand sanitizer X as a positive control, F0 as
negative control and hand sanitizer formula F1 (0,5%
extract), F2 (1% extract) and F3 (1,5% extract). This
entire prepare process done in a sterile inside leminer
airflow to prevent contamination of the media. Analysis
of the quality of hand sanitizer includes organoleptic, the
measurement of pH, homogeneity test and dispersive
power test.
RESULT AND DISCUSSION
The extraction of pineapple peel obtained using
maceration for 6 days with ethanol is about 1.2 L. After
concentrated become as much as 16.7302 g, or 5.5767%.
These concentrated extract then used to phytochemical
analysis, preparation of hand sanitizer and antibacterial
activity test. Results of phytochemical screening
pineapple peel samples are presented in Table 2.
This is according to research from Yeragamreddy
et al., (2013) that pineapple peel contains tannins,
saponins and flavonoids. This compounds (flavonoids,
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Figure 1. UV-Vis spectra of pineapple peel extract by
adding a sliding reagent a (metanol), b
(NaOH), c (AlCl3), d. (AlCl3 + HCl)
Table 3. Wavelength shift of UV-Vis analysis
tannins and saponins) has aromatic hydroxyl groups
which act as antibacterial. Analysis using UV-Vis
measured in the λ 230-530 nm. UV-Vis spectrum
extract of pineapple peel by adding a sliding reagent can
be seen in Figure 1.
Based on the results of UV-Vis ethanol extract of
the pineapple peel, obtained two absorption peaks
indicating flavonoid compounds that is peak II at 291 nm
and peak I at 361 nm. According to Markham
(1988), the maximum absorption is typically of
flavonoid compounds groups of flavanones and
dihidroflavonolwhich has an absorption maximum
between 275-295 nm on band II and 300-330 nm on
band I. Analysis is continued by adding a sliding reagent
that will clarify the structure of flavonoids.
The addition of NaOH 2M sliding reagent
causes a bathochromic shift towards higher
wavelengths 32 nm in the band II indicate the possibility
of ortho dihidroksil group on the ring A. Whereas a
wavelength shift of 50 nm on the band I showed hydroxy
 International Conference on Mathematics, Science, and Education 2016 (ICMSE 2016)
Figure 2. IR spectrum analysis results
Gambar 3. Clear zone of Escherichia coli
Figure 4. Clear zone of Staphylococcus aureus
groups in ring B in position 3, 4'-OH. The addition of
NaOH 2M sliding reagent is read again after 5 minutes
and occur as much as 4 nm bathochromic shift in the
band II underscoring their ortho dihidroksil group located
on the ring A.
Sliding reagent AlCl3 that was added has been
shifted the maximum absorbance towards the higher
wavelength to the 26 nm in the band II that showed ortho
dihidroksil on ring A (6.7 or 7.8) (Markham, 1988).
Whereas bathochromic shift in the band I by 62 nm
enables the ortho hydroxyl group at C-4 'and C-5'.
The addition of a sliding reagent AlCl3-HCl
showed a shift in the band I at 49 nm, there might be a
hydroxyl group at C-3 which can form acid resistant
complexes with ketone group (Markham, 1988).
Existence of the hydroxyl group at C-3 indicates that
flavonoids in the extract is not a group of flavonon but a
group of dihidroflavanol (Markham, 1988). UV-Vis
analysis with the addition of a sliding reagents obtained
allegation that the compounds contained in extracts of
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pineapple peel is a compound derivative of
dihidroflavonol.
To reinforce the notion that the spectrum appears
in UV-Vis analysis is a flavonoids compound, then we
conducted the analysis using infrared spectroscopy.IR
spectrum analysis results can be seen in Figure 2, and the
IR spectrum interpretations data presented in Table 3.
Based on the data in Table 3 above is
estimated functional groups of the building
compounds of pineapple peel extract is as follows:
wide absorption with moderate intensity at wave
number 3365 cm-1 that is suspected absorption of O-H
group. Absorbtion of C-H aliphatic sharp and low
appears at wave number 2925 cm-1 and 2859 cm-1. It is
supported by the results of research by Akbar (2010) that
the absorption at wavenumber 2927 cm-1 shows the
absorption C-H in the group at C-H aliphatic. The
existence carbonyl (C=O) as a common trait of
flavonoid (Sukadana, 2010) is indicated by the
-
presence of absorption at the wave number 1725 cm
1
. Absorption of C=C aromatic group appeared on the
wave of 1642 cm-1. Absorption of C=C aromatic group
appeared on the wave of 1642 cm-1. C-H group
appeared at 1341 cm-1. Absorption C-O in phenol
compounds produce a strong band in the 1050- 1300
cm-1 (Skoog et al., 1998), and on the analysis of C-
O absorption appears at wave number 1232 cm-1 with
the absorption medium, and with a sharp absorption in
1065 cm-1. The existence of functional groups O-H, CH
aliphatic, aromatic C=C and C-O indicates a flavonoid
compound. This was confirmed by the results of research
conducted by Akbar (2010) on the results of
infra-red spectrum presence of functional groups O-
H, C=O, C-O, C=C aromaticand C-H aliphatic
support that extract positive a flavonoid compound.
The results of antibacterial activity of pineapple
peel extract showed good results on both bacteria that is
Escherichia coli and Staphylococcus aureus clear zone
formed can be seen in Figure 3 and Figure 4.
Test results of antibacterial activity against
Escherichia coli are presented in Table 4, and the test
result of antibacterial activity against Staphylococcus
aureus, are presented in Table 5.
Antibacterial activity resulted inhibition zone
for pineapple peel extract is 16.5 mm for
Escherichia coli and 17 mm for Staphylococcus
aureus, a positive control that hand sanitizer Cussons
CAREX® produce inhibition zone 9 mm for Escherichia
 International Conference on Mathematics, Science, and Education 2016 (ICMSE 2016)
Table 4. Antibacterial activity against Escherichia coli
Table 5. Antibacterial activity against Staphylococcus
aureus
coli and 10,5 mm for Staphylococcus aureus, hand
sanitizer formula (F1) with 0.5% extract of pineapple
peel produce inhibition zone of 9 mm for Escherichia
coli and 10 mm for Staphylococcus aureus, hand
sanitizer formula (F2) with 1% pineapple peel extract
produce inhibition zone of 13 mm for Escherichia coli
and 15 mm for Staphylococcus aureus, hand sanitizer
formula (F3) with 1.5% extract of pineapple peel
produce inhibition zone of 15 mm for Escherichia coli
and 15.5 mm for Staphylococcus aureus. While the
negative control that hand sanitizer formula without
extract of pineapple peel is not found any clear
zone in isolates of Escherichia coli and Staphylococcus
aureus, this means a basic ingredient in hand sanitizers
really do not have antibacterial activity, but the
pineapple peel extracts as active ingredients proven
effective as an antibacterial agent,is even more
effective than positive controls were tested. Getting
bigger concentration of extract added, inhibitory zone
also will increase. This proves the increased
effectiveness in inhibiting bacteria along with the
addition of the extract in the formulation. Pelczar and
Chan (1986) says that the higher concentration of an
antimicrobial agent, the cells of microorganisms will be
more rapid killed or stunted.
Preparations hand sanitizers that have been tested
for antibacterial activity is then analyzed through
organoleptic test, homogeneity, pH and dispersive power
to determine the physical quality of hand sanitizer and
selected formulation with the best quality. The results
of organoleptic test that includes observation of form,
color, and odor in the preparation of hand sanitizer are
presented in Table 6.
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Table 7. Results of the organoleptic test
Table 8. Results of pH measurement
Result of the organoleptic test on the fourth hand
sanitizer formulations have the same form that is viscous.
But the colors on each formulationis different.Hand
sanitizer without addition of pineapple peel extract (F0)
is colorless. Meanwhile the hand sanitizer formulation
with extracts of pineapple peel is yellow, and color
density increases with addition of extract. In observation
of the smell of formulation, F0, F1, F2 and F3 was
fragrance it comes from addition of perfume. Results of
pH measurement has been done every week in one
month period is served in Table 7.
pH measurement is done at weeks 1,2,3 and 4
show the different results in each formula. Formula
with 0% extract of pineapple peel (F0) showed neutral
pH at weeks 1st and 2nd, then stabilized at pH = 6 on
weeks 3rd and 4th. Formula hand sanitizer with 0.5%
pineapple peel extracts (F1) experienced an increase in
the pH of
6 to 7 and then fell back into a stable at pH = 6.
Formula hand sanitizer with 1% extract (F2) is stable at
pH = 6 for 3 weeks, but then decreased at week 4th at pH
= 5. Formula hand sanitizer with 1.5% extract showed a
pH value = 6 in the first week and then stable at pH = 5
in next week. pH test for 4 weeks showed a decrease pH
along with the addition of pineapple peel extract, it’s
because the extract of pineapple peel pH is acidic (pH =
4) so increased of extract, levels of pH decreases. But all
formulations with the addition of pineapple peel extract,
there is no lower than the pH of the skin range is 4,5-6,5
(SNI, 1992; Wilkinson, 1982: Shu, 2013). That is, the
preparation has a good pH value and still safely applied
to the skin.
Results of homogeneity test of hand sanitizer is
not found a clot or coarse grain at all four
formulations. This means that all formulas have been
homogeneous or perfectly mixed so the quality of hand
sanitizer in this case can be said is good because it has
met the quality requirements which the preparations
 International Conference on Mathematics, Science, and Education 2016 (ICMSE 2016)
must show a homogeneous composition and no visible
coarse grains (Ditjen POM, 1979). While the results of
dispersive power test of hand sanitizer with pineapple
peel extract also meets the standards dispersive power gel
that is more than 4 cm (Garg et al., 2002)
The results of hand sanitizer quality test includes
organoleptic tests, the measurement of pH, homogeneity
test and dispersive power test, hand sanitizer with
extracts of pineapple peel and hand sanitizer without
the extract of pineapple peel
shows a good results and meet the quality
requirements of hand sanitizer. However, the best among
fourth formulas are hand sanitizer F3 with 1.5% extract
of pineapple peel because it has the greatest inhibition
against Staphylococcus aureus and Escherichia coli than
other formulas of hand sanitizer. As well as testing the
quality of hand sanitizer, F3 still qualify a good quality.
CONCLUSION
Ethanol extract of pineapple peel with maceration
method proved able to inhibit the growth of bacteria
Escherichia coli and Staphylococcus aureus very well,
including those were applied as a hand sanitizer. Hand
sanitizer formulation with pineapple peel extract at
concentrations 0.5%, 1% and 1.5% extract produces
inhibition zone against Escherichia coli that is 9 mm, 13
mm and 15 mm, while against Staphylococcus aureus
produces inhibition zone of 10 mm, 15 mm and 15.5
mm. Getting bigger concentration of extract added,
inhibitory zone also will increase. Quality hand sanitizer
by adding pineapple peel extract have passed
organoleptic test, pH, homogeneity and dispersive power.
The best hand sanitizer formulas are hand sanitizer F3
because it has the greatest inhibition than other formulas
and the quality test of hand sanitizer F3 still meets the
standard of hand sanitizer.
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