Available via license: CC BY-NC 4.0
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J. Tek. Kim. Ling. 2023, 7 (2), 114-122
p-ISSN : 2579-8537, e-ISSN : 2579-9746
http://jurnal.polinema.ac.id/index.php/jtkl
DOI: http://dx.doi.org/10.33795/jtkl.v7i2.3737
*Corresponding author: Wa Ode Cakra Nirwana
Received : August 6, 2023
Department of Chemical Engineering, University of Brawijaya
Accepted : October 19, 2023
MT. Haryono No. 167, Malang, 65145, Indonesia
E-mail: cnirwana@ub.ac.id
Potential of Citronella Oil and Gum Rosin as
Antimicrobial Agents in Floor Cleaner Liquid Against
Salmonella thypi
Wa Ode Cakra Nirwana1,2,*, Chandrawati Cahyani1,2, Vivi Nurhadianty1,2
1Department of Chemical Engineering, Faculty of Engineering, University of Brawijaya, MT. Haryono No. 167,
Malang, 65145, Indonesia
2Institute of Essential Oil, University of Brawijaya, Gedung Senat 1st floor, Veteran, Malang 65145, Indonesia
ABSTRACT
Some floor cleaner products on the market contain antimicrobial agents that are harmful to humans and the
environment, such as benzalkonium chloride. Therefore, searching for a new antimicrobial agent that is
relatively safe is a significant challenge. In this study, we investigated the best formula for making floor cleaner
using citronella oil and gum rosin as antimicrobial agents. Its antimicrobial activity against Salmonella thypi
was investigated and compared with that of a commercial floor cleaner containing benzalkonium chloride. Five
concentrations and ratios of citronella oil and gum rosin were investigated. The quality of floor cleaner in terms
of pH range and the stability of the emulsion in hard water was assessed according to SNI 06-1842-1995. The
results showed that all formulas met the pH range required by SNI, except for formulas containing citronella oil
alone. The emulsion in hard water was stable for all formulas. The antimicrobial activity increased with
increasing concentrations of antimicrobial agents. Citronella oil exhibited higher antimicrobial activity than gum
rosin. Benzalkonium chloride showed higher microbial activity than gum rosin but lower than citronella oil. The
combination of citronella oil and gum rosin showed higher microbial activity by 1.1–2.4 times than formulas
containing benzalkonium chloride. The optimum floor cleaner formula (a concentration of citronella oil and
gum rosin of 2% with a ratio of 1:1) has an inhibition zone diameter of 22.2 mm.
Keywords: antimicrobial, citronella oil, floor cleaner, gum rosin.
1. INTRODUCTION
Currently, many floor cleaning products
offer various advantages such as being able
to clean floors, make floors shinier, smell
fresh and last longer, and as antimicrobial.
Of the many advantages offered, the ability
to act as an antimicrobial is the main
attraction for consumers. However, some of
the antimicrobial agents used are hazardous
to humans and the environment.
In Indonesia, floor cleaning products
generally use benzalkonium chloride,
cresylic acid, ethoxylated alcohol, and pine
oil as antimicrobial agents (personal
observation). Benzalkonium chloride is
dangerous because it is corrosive, causes
severe skin burns and eye damage, may
cause respiratory irritation, and is very toxic
to aquatic life with long-lasting effects [1].
Cresylic acid is toxic, corrosive, cause
severe burns if in contact with skin, and can
affect the central nervous system and other
organs (lungs, liver, kidneys, and eyes) [2].
Ethoxylated alcohol is corrosive, harmful if
swallowed and in contact with skin and
eyes, and very toxic to aquatic life [3].
Meanwhile, pine oil is relatively safer and
does not have the potential to cause chronic
health problems, but it can cause irritation if
in contact with the eyes and sensitive skin
[4].
As previously explained, with the negative
effects of antimicrobial agents used in
commercial floor cleaning products on
human health and the environment, using
natural ingredients that are relatively safer in
Nirwana, et al./ Jurnal Teknik Kimia dan Lingkungan, Vol. 7, No. 2, October 2023
115
household products, particularly floor
cleaners, is highly important. Natural active
ingredients that have the potential to be used
as antimicrobials in floor cleaners liquid are
citronella oil and gum rosin.
The main components of citronella oil are
citronellal, geraniol and citronellol [5].
These components have antibacterial
properties that can inhibit the growth of
bacteria that cause food spoilage and
diseases such as Brochothrix thermosphacta,
Escherichia coli, Listeria innocua, Listeria
monocytogenes, Pseudomonas putida,
Salmonella typhimurium, and Shewanella
putrefaciens [6–9].
Gum rosin is obtained from the distillation
residue of the sap of pine trees [10]. Its
constituent components vary, strongly
influenced by the species of pine,
environment, plant age, geographic origin,
resin-tapping techniques, etc. [11–13]. The
main components of gum rosin are diphenic
acid, mainly abietic acid, isopimaric acid,
laevoabietat acid, and pimaric acid [10].
Several studies have shown that abietic acid
has a function as an antibacterial that causes
disease [14,15].
To date, research regarding the production
of floor cleaner liquid using natural
ingredients as antimicrobial agents has been
widely explored. The antimicrobial agents
used include lerak fruit [16], mangosteen
peel [17], kaffir lime leaf oil, lemongrass oil,
and its hydrosol [18]. To the best of the
authors’ knowledge, study of the use of
citronella oil and gum rosin as antimicrobial
agents in a floor cleaner product has not
been investigated. In this work, we
investigated the optimum formula of
citronella oil and gum rosin as antimicrobial
agents in floor cleaner liquid against
Salmonella thypi. Its antimicrobial activity
was then compared with that of a
commercial floor cleaner that uses
benzalkonium chloride as the antimicrobial
agent.
2. MATERIALS AND METHODS
2.1. MATERIALS
Citronella oil was obtained from Essential
Oil Institute, University of Brawijaya,
Malang, Indonesia. Sodium hydroxide, gum
rosin, (MES), nutrient agar, and nutrient
broth were purchased from local suppliers.
2.2. PRODUCTION OF FLOOR
CLEANER LIQUID USING
CITRONELLA OIL AND GUM
ROSIN AS ANTIMICROBIAL
AGENTS
Five concentrations of antimicrobial agent
i.e. 1–5% w/v (mass of citronella oil and
gum rosin per volume of aquadest) were
evaluated. While the ratio of citronella oil to
gum rosin was 1 : 0; 0 : 1; 1 : 1; 2 : 1; 1 : 2
(w/w). The experiments were carried out
using a completely randomized design
method; hence, the number of formulas was
25.
The flowchart for making floor cleaner
liquid is shown in Figure 1. NaOH (0.2 mass
of gum rosin) was dissolved in aquadest.
Subsequently, gum rosin and the remaining
aquadest were added, stirred, and heated at
75 oC for 15 min. After cooling down to
room temperature, MES (0.3 mass of gum
rosin) was added. After it dissolves
completely, citronella oil was added and
stirred until homogenous.
2.3. MICROORGANISM AND
INOCULUM PREPARATION
Salmonella thypi used for antimicrobial
activity testing was purchased from the
Faculty of Medicine, University of
Brawijaya. The strain was grown on nutrient
agar and incubated at 37 oC overnight. The
inoculum preparation procedure refers to the
standard procedure by the Clinical and
Laboratory Standards Institute (CLSI) [19]
with a slight modification. Four colonies
were transferred to 10 mL of nutrient broth
and then incubated at 37 oC for 12 h. The
turbidity of the cell suspension was adjusted
with sterile nutrient broth to obtain turbidity
comparable to the 0.5 McFarland standard
Nirwana, et al./ Jurnal Teknik Kimia dan Lingkungan, Vol. 7, No. 2, October 2023
116
(containing approximately 1.5 x 108
CFU/mL).
2.4. ASSESSMENT OF THE QUALITY
OF FLOOR CLEANER
The quality of the floor cleaner in terms of
pH and stability of emulsion in hard water
was assessed according to SNI 06-1842-
1995 [20]. The pH was measured
periodically for 11 days using a pH meter
(OHAUS, Starter 300, USA). The
assessment of the stability of the emulsion in
hard water was carried out by dissolving the
floor cleaner in hard water with ratios of
5:100 and 1:100 (v/v). Subsequently, the
stability of emulsion was observed visually
after 6 hours. Stability refers to the absence
of precipitation or separate layers. For floor
cleaners containing phenol and its
derivatives, the emulsion in hard water must
be stable while for other compounds, it does
not form an emulsion (SNI 06-1842-1995).
*The composition of gum rosin and citronella oil was
adjusted according to the variables investigated in
this work.
Figure 1. Procedures for making floor
cleaner liquid with antimicrobial compounds
of citronella oil and gum rosin.
2.5. ANTIMICROBIAL ACTIVITY
TESTING
The protocol of antimicrobial activity testing
using the disk diffusion method refers to
CLSI standards with slight modification
[19]. A 0.1 mL of bacterial suspension was
spread on the surface of a nutrient agar using
a bacterial cell spreader. The disks were
immersed in the floor cleaner liquid then
placed on the inoculated agar surface and
incubated at 37 °C. The antibacterial activity
was evaluated by measuring the diameter of
inhibition zone of the tested bacteria using a
ruler as depicted in Figure 2, then recorded
as an average, which was expressed in
millimeters. As a comparison, the
antimicrobial activity of a commercial floor
cleaner liquid containing benzalkonium
chloride was also conducted to compare
their effectiveness against the strain. The
concentration of benzalkonium chloride in
the floor cleaner liquid was adjusted to the
concentrations of the antimicrobial agent
specified in this work. The experiments
were conducted in triplicate.
Figure 2. Method of inhibition zone
measurement.
A=disc paper; B= inhibition zone; C=petri
dish
3. RESULTS AND DISCUSSION
3.1. pH TESTING
According to the Indonesian standard for
floor cleaner products (SNI-06-1842-1995),
the pH must be in the range of 6–11. As
depicted in Figure 3, the pH of floor
cleaners varied. The pH was less than 6
when the floor cleaner contained citronella
oil alone as the antimicrobial agent. In
contrast, using gum rosin resulted in a pH of
around 8–10. A combination of citronella oil
and gum rosin with various concentrations
and ratios produced floor cleaners with a
pH of 7–10. Observation of pH changes for
11 days showed that the pH of floor cleaners
was relatively stable. As a comparison, the
pH of the commercial floor cleaner
A
B
C
Nirwana, et al./ Jurnal Teknik Kimia dan Lingkungan, Vol. 7, No. 2, October 2023
117
containing benzalkonium chloride was 10
(data not shown). These results show that
the floor cleaners produced in this work met
the pH standards required by SNI, except for
formulas that used citronella oil alone. This
is because the pH of citronella oil used in
this study was 5.
3.2. TESTING OF EMULSION
STABILITY IN HARD WATER
The purpose of this test is to observe the
stability of floor cleaner emulsion in hard
water. Two ratios of floor cleaner to hard
water were observed, namely 1:100 and
5:100. Table 1 shows the visualization of
floor cleaners in hard water after 6 h with a
total concentration of citronella oil and gum
rosin of 5% (other concentrations are not
shown for simplification). The results show
that samples not containing gum rosin did
not form emulsions. As previously
described, according to SNI 06-1842-1995,
for floor cleaners containing phenol and its
derivatives, the emulsion in hard water must
be stable, while for other compounds, it does
not form an emulsion. The main components
of gum rosins are abietic acid, isopimaric
acid, laevoabietat acid, and pimaric acid
which are categorized as phenol derivative
compounds, thereby forming an emulsion in
hard water. In contrast, the main
components of citronella oil are citronellal,
geraniol, and citronellol which are
categorized as non-phenol compounds.
Therefore, it did not form an emulsion in
hard water.
After 6 h of observation, all samples showed
there was no precipitation, floc formation, or
separated layer. It indicates that the
emulsion of floor cleaners containing
citronella oil and gum rosin is stable and
homogeneous in hard water.
4
6
8
10
12
4
6
8
10
12
0 2 4 6 8 10 12
0 2 4 6 8 10 12
4
6
8
10
12
1%; 0:1
1%; 1:0
1%; 1:1
1%; 1:2
1%; 2:1
pH
2%; 0:1
2%; 1:0
2%; 1:1
2%; 1:2
2%; 2:1
3%; 0:1
3%; 1:0
3%; 1:1
3%; 1:2
3%; 2:1
pH
4%; 0:1
4%; 1:0
4%; 1:1
4%; 1:2
4%; 2:1
Day
5%; 0:1
5%; 1:0
5%; 1:1
5%; 1:2
5%; 2:1
pH
Day
Figure 3. Profile of pH changes in floor cleaners containing citronella oil and gum rosin.
Legend represents the total concentration of citronella oil and gum rosin in floor cleaner; ratio
of citronella oil to gum rosin (w/w).
Nirwana, et al./ Jurnal Teknik Kimia dan Lingkungan, Vol. 7, No. 2, October 2023
118
Table 1. Visualization of homogeneity of floor cleaners in hard water after 6 h with a total
concentration of citronella oil and gum rosin of 5%.
Ratio of
floor
cleaner to
hard water
Ratio of citronella oil to gum rosin
0:1
1:0
1:1
1:2
2:1
1:100
5:100
3.3. ANTIMICROBIAL ACTIVITY
TESTING
Salmonella thypi was chosen as the tested
bacteria because this is one of the bacteria
that is commonly found on the floor. The
antimicrobial activity of citronella oil and
gum rosin was assessed according to the size
of the clear zone (inhibition zone) around
the disc paper. A larger zone of inhibition
around an antimicrobial-containing disk
indicates that the bacteria are more sensitive
to the antimicrobial in the disk and vice
versa (Figure 4).
Figure 5 shows the effect of concentrations
and ratios of citronella oil and gum rosin in
floor cleaner on the inhibition zone against
S. thypi. In general, the diameter of the
inhibition zone increased with increasing
concentrations of antimicrobial agents. Floor
cleaners containing citronella oil alone
showed a larger inhibition zone of around
2.5–3.1 times than floor cleaners containing
gum rosin alone under the range
concentrations observed in this work. It
indicates that S. thypi is more sensitive to
citronella oil than gum rosin. The
combination of citronella oil and gum rosin
as antimicrobial agents increased the
inhibition of S. thypi growth. A higher ratio
of citronella oil increased the inhibition
zone. The same phenomenon was also
observed for gum rosin. The highest
inhibition zone for each ratio of citronella
oil : gum rosin was 0:1 = 7.3 mm; 1:0 = 21.1
mm; 1:1 = 22.2 mm; 1:2 = 22.8 mm; and 2:1
= 19.3 mm, respectively, achieved at
concentrations of 3%, 2%, 2%, 5%, and 2%.
Citronella oil is well known to have good
antimicrobial efficacy since it contains
citronellal, geraniol, and citronellol which
have been described as antiseptic,
antibacterial, and antifungal.
Figure 4. Clear (inhibition) zone around the
disc paper on the antimicrobial activity
testing.
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119
1 2 3 4 5
0
5
10
15
20
25
30
35
40
Average diameter of the inhibition zone (mm)
Total concentration of active ingredients of
citronella oil and gum rosin (%)
CO:GR = 0:1
CO:GR = 1:0
CO:GR = 1:1
CO:GR = 1:2
CO:GR = 2:1
Figure 5. Effect of concentrations and ratios
of citronella oil and gum rosin in floor
cleaners on the inhibition zone against S.
Thypi.
CO=citronella oil; GR=gum rosin
The mechanism of action of these
compounds occurs by increasing the
membrane fluidity and permeability of the
microorganisms, resulting in cellular
disruption or lysis [5]. Gum rosin is a
diterpenoid mainly composed of diphenic
acid mainly abietic acid, isopimaric acid,
laevoabietat acid, and pimaric acid [10].
Studies showed that diterpenoid compounds,
particularly abietic acid have antiviral,
antibiotic, and antifungal properties [21]. It
is also effective against bacteria, particularly
gram-positive bacteria [22]. This might
explain why the antimicrobial activity of
gum rosin against the strain was relatively
low in this study since S. thypi is a gram-
negative bacteria. The antimicrobial
mechanism of diterpenoids, particularly
abietic acid, might be attributed to the
carboxylic group, which interacts with the
lipid component of the bacterial cellular
membrane, disrupting the lipophilic cell
membrane. As a result, the cell membranes
undergo lysis [14,21].
To assess the effectiveness of floor cleaners
formulated in this study, the antimicrobial
activity of the commercial floor cleaner
containing benzalkonium chloride was then
investigated. As presented in Figure 6, for
all concentrations, the commercial floor
cleaner exhibited higher microbial activity
against S. thypi by 1.4–2.2 times compared
to floor cleaners containing gum rosin alone.
In contrast, its microbial activity was 27–56
% lower compared to floor cleaner
containing citronella oil alone. All formulas
using a combination of citronella oil and
gum rosin showed higher microbial activity
by 1.1–2.4 times than formulas using
benzalkonium chloride. The highest
microbial activity was achieved at the
formula of 5% of citronella oil and gum
rosin with a ratio of 1:2. Under these
conditions, the average diameter of the
inhibition zone was 22.8±1.1 mm, not
significantly different compared to the
formula with the concentration of citronella
oil and gum rosin of 2% with a ratio of 1:1.
According to the work of Söderberg et al.
(1990), they found that abietic acid could be
chemically converted in agar plates seeded
with Escherichia coli and Staphylococcus
aureus. Abietic acid was oxidized during the
experiment to the most stable abietic type of
acid, namely dehydroabietic acid, which had
a slight antibacterial effect against some
strains of Bacillus subtilis but no effect
towards S. aureus, gram-negative bacteria,
or fungi. This finding might explain why in
this study, the increasing gum rosin ratio did
not increase the inhibition zone significantly
against S. thypi [23].
The inhibition zone for a floor cleaner
containing 2% citronella oil and gum rosin
at a ratio of 1:1 was 22.2 ± 1.2 mm.
Meanwhile, when using 5% citronella oil
and gum rosin at a ratio of 1:2, the zone of
inhibition was 22.8 ± 1.1 mm. It shows that
increasing the concentration of antimicrobial
agents by 2.5 times results in an increase in
the inhibition zone of only 2%. Therefore, a
concentration of antimicrobial agents of 2%
with a ratio of 1:1 is considered the optimum
condition.
Nirwana, et al./ Jurnal Teknik Kimia dan Lingkungan, Vol. 7, No. 2, October 2023
120
Figure 6. Comparison of the inhibition zone diameter of the floor cleaners containing
citronella oil and gum rosin with the commercial floor cleaner containing benzalkonium
chloride.
4. CONCLUSION
In this work, the production of floor cleaner
liquid using citronella oil and gum rosin as
antimicrobial agents has been demonstrated.
Floor cleaners have a relatively stable pH
and emulsion in hard water. Floor cleaners
containing citronella oil and gum rosin
developed in this work (concentration 2%,
ratio 1:1) had antimicrobial activity against
Salmonella thypi 2.1 times higher than the
commercial floor cleaner containing
benzalkonium chloride (concentration 2%).
Citronella oil and gum rosin have great
potential as antimicrobial agents for floor
cleaning products.
ACKNOWLEDGMENT
Authors would like to thank the Faculty of
Engineering, University of Brawijaya for
financially supporting this research.
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