Formulation and Evaluation of Citronella Oil (Cymbopogon nardus (L.) Rendle) Cream for Acne Treatment

Abstract

Acne is a prevalent skin disorder that affects 80–85% of teenagers globally. Citronella oil is one of the natural compounds that has been known to potentially treat acne, but its application is limited due to its greasiness and uncomfortable sensation on skin. This study aimed to optimize the concentration of cetyl alcohol as a viscosity enhancer that can produce a physically stable cream preparation and to evaluate the antibacterial activity of the optimized formula against Propionibacterium acnes. Following the emulsification process, the physical characteristics of the five cream formulas, including organoleptic, homogeneity, emulsion type, pH, adhesion, specific gravity, viscosity, and stability were evaluated. Formula 3 (F3), cream formulation with 6% cetyl alcohol having physical characteristics of a white homogeneous cream with a pronounced lemongrass scent, emulsion type o/w, pH of 6.30 ± 0.02, adhesion of 16.85 ± 0.58 s, a specific gravity of 1.031 ± 0.009 g/mL, and viscosity of 4418 ± 182 mPas was chosen as the optimized formula. F3 was then subjected to antibacterial testing against P. acne and showed a 9.35 mm inhibition zone. Cream-based citronella oil, therefore, becomes a promising preparation for acne treatment.

Full text

INTRODUCTION
Acne vulgaris, or acne, is a highly prevalent skin condition
affecting 80 –85% of adolescents worldwide.1 Acne is a
disease that aects which can be caused by excess sebum, the
Propionibacterium acnes infection, keratinocyte shedding,
hormones, genes (heredity), the environment, and medication
or cosmetic use.2,3 Acne typically develops on a place where
it has a high density of sebaceous glands such as face, chest,
upper back, and upper arms.4 The sebum can be caught in
the keratin and hardened. As a result of the trapped sebum,
P. acnes, a member of the skin microora, can grow and
eventually produce comedones, inammatory acne, papules,
and even scar tissue.3
Acne therapy is necessary since it can have negative
social eects, including depression, anxiety, and low self-
esteem.5 Topical acne medications such retinoids, benzoyl
peroxide, azelaic acid, keratolytic, and alpha-hydroxy acids
are commonly caused skin irritation or stinging, while the use
of topical antibiotics in recent years has become increasingly
ineffective due to the emergence of antibiotic-resistant
strains.6,7 Other alternatives that can provide relatively lower
side eects are needed to treat acne. Natural ingredients are
used more frequently as natural antibacterials since they have
fewer side eects than conventional dr ugs.8
Lemongrass (Cymbopogon nardus (L.) Rendle), one of the
many herbal plants from Indonesia, has been studied for its
potential to combat acne-causing bacteria, including P. acn es
and Staphylococcus epidermidis in-vitro.9-10 Lemongrass
contains citronella oil with citronellal, geraniol, and citronellol
as active substances.11 These compounds remove the intra-
cellular hydrogen ions, causing immobility and eventually cell
death in bacteria.12 However, citronella oil cannot be applied
directly to the skin since it may lead to irritation.13 Top i c a l
dosage forms are favored for treating acne due to their ease
of use.14 Creams are semi-solid topical formulations in the
form of a water-in-oil (w/w) or oil-in-water (o/w) emulsion.15
Cream formulations have advantages such as the convenience
of application, good spreadability, and release of active
ingredients into the skin.16
Therefore, this research was aimed to optimize cetyl
alcohol concentration as a viscosity enhancer in anti-acne
cream preparations of citronella oil. Cetyl alcohol was selected
as a thickener because it has emollient properties, an occlusive
ba r r ie r on th e skin, an d hel ps to keep the skin hydrat e d.17-18 The
preparations were evaluated for their physical stability and the
ABSTRACT
Acne is a prevalent skin disorder that aects 80–85% of teenagers globally. Citronella oil is one of the natural compounds that
has been known to potentially treat acne, but its application is limited due to its greasiness and uncomfortable sensation on skin.
This study aimed to optimize the concentration of cetyl alcohol as a viscosity enhancer that can produce a physically stable cream
preparation and to evaluate the antibacterial activity of the optimized formula against Propionibacterium acnes. Following the
emulsication process, the physical characteristics of the ve cream formulas, including organoleptic, homogeneity, emulsion
type, pH, adhesion, specic gravity, viscosity, and stability were evaluated. Formula 3 (F3), cream formulation with 6% cetyl
alcohol having physical characteristics of a white homogeneous cream with a pronounced lemongrass scent, emulsion type
o/w, pH of 6.30 ± 0.02, adhesion of 16.85 ± 0.58 s, a specic gravity of 1.031 ± 0.009 g/mL, and viscosity of 4418 ± 182 m
Pas was chosen as the optimized formula. F3 was then subjected to antibacterial testing against P. acne and showed a 9.35
mm inhibition zone. Cream-based citronella oil, therefore, becomes a promising preparation for acne treatment.
Keywords: Cream, Emulsion, Citronella oil, Propionibacterium acnes, Viscosity enhancer.
International Journal of Drug Delivery Technology (2023); DOI: 10.25258/ijddt.13.1.67
How to cite this article: Nareswari TL, Vrince FO, Syatri E. Formulation and Evaluation of Citronella Oil (Cymbopogon
nardus (L.) Rendle) Cream for Acne Treatment. International Journal of Drug Delivery Technology. 2023;13(1):418-422.
Source of support: Nil.
Conict of interest: None
Formulation and Evaluation of Citronella Oil (Cymbopogon nardus (L.)
Rendle) Cream for Acne Treatment
Tantri L. Nareswari*, Fidela O. Vrince, Erga Syatri
Department of Pharmacy, Faculty of Science, Institut Teknologi Sumatera, Lampung, Indonesia
Received: 09th January, 2023; Revised: 16th January, 2023; Accepted: 08th February, 2023; Available Online: 25th March, 2023
RESEARCH ARTICLE
*Author for Correspondence: tantri.nareswari@fa.itera.ac.id

Formulation of Citronella Oil Cream for Acne Treatment
IJDDT, Volume 13 Issue 1, January - March 2023 Page 419
optimum formula was then subjected to an in-vitro antibacterial
test against P. acnes. This study is expected to produce stable
and good-quality citronella oil cream compositions, eectively
inhibiting the acne-causing bacteria P. a cnes.
MATERIALS AND METHODS
Materials
The materials used in this study included citronella essential
oil (PT. Rumah Atsiri Indonesia) derived from citronella plants
(C. nardus (L.) Rendle) from Tawangmangu, Central Java.
Tween 60, Span 60, cetyl alcohol, stearyl alcohol, sodium
benzoate, glycerin, alpha-tocopherol, and aquadest are all
pharmaceutical grade. Blood base agar and 0.9% NaCl was
sterilized, and P. acnes ATCC 11827 was purchased from
University of Indonesia.
Citronella Oil Cream Formulation
Optimization was carried out on cetyl alcohol as a viscosity
enhancer with different concentrations (2, 4, 6, 8, 10%).
Cream formulation was prepared by emulsication method as
described by Lohani et al. with some modications.19 In a brie f,
the oil phase (1.6% span 60, cetyl alcohol, 2% stearyl alcohol,
and 0.05% alpha-tocopherol) was mixed in a mechanical
overhead stirrer at 100–200 rpm in the temperature of 60 C.
The aqueous phase consisted of 3.4% tween 60, 3% glycerin,
0.2% sodium benzoate and water. The water phase was added
to the oil phase at 100–200 rpm at 60˚C and mixed continuously
until semi-solid consistency was achieved. The temperature
is then lowered to <30  C. Citronella oil is gradually added
until uniform.
The cream formed was then evaluated for its physical
preparation, including organoleptic, homogeneity, emulsion
type, pH, adhesion, specic gravity, viscosity, and stability
test. The optimized formula was selected for an antibacterial
activity test.
Organoleptic Test
Organoleptic evaluation of citronella oil cream was conducted
by visual inspection of the formulation, including appearance,
consistency, odor and color.20
Homogeneity Test
Testing the homogeneity of citronella oil cream preparations
was carried out by taking 1-gm of cream at the top, middle and
bottom, then smearing it on an object glass. Cream preparations
can be said to be homogeneous if there are no visually visible
coarse grains.21
Emulsion Type Test
The type of emulsion is determined by adding water on a
certain amount of cream. The cream that can be diluted is the
O/W type, whereas the cream that cannot be diluted is the
W/O type.22
pH test
Sample was weighed as much as 0.5 gm, dissolved in 50 mL
of distilled water, and tested with a pH-meter.
Adhesion Test
The adhesion test was carried out by attaching sucient cream
to two glass objects, then pressing it with a load of 0.5 kg and
then adding a load weighing 20 gm for 5 minutes. Furthermore,
the time when the cream was released from the two glass
objects was recorded.23
Specic Gravity Test
The specic gravity test was carried out using a pycnometer.24 A
25 mL empty and clean pycnometer is weighed rst, then 25 mL
of cream is added and closed, then weighed. Do the same for
the water as a control. Determination of specic gravity with
the formula (1).
(1)
where ρ1 = specic gravity of water; ρ2 = specic gravity of
cream; W2 = pycnometer with cream; W1 = pycnometer with
water; W0 = empty pycnometer.
Viscosity Test
Viscosity testing was carried out using a viscometer germany
Rotavisc HiVi I (Ika) using spindle number 10 at 100 rpm.
Three replications of samples were measured.
Stability Study
The ve formulations were subjected to a room temperature
stability test for 28 days and three freeze-thaw cycles. This test
was carried out at room temperature 25˚C days 0, 1, 7, 14, 21, 28.
Freeze-thaw storage temperature was -18˚C for 24 hours
and 25˚C for 24 hours for 1 cycle.25 The physical properties
(organoleptic, homogeneity, adhesion, pH, and viscosity) were
evaluated at each time point.
Optimized Formula Determination
The optimized formula derived from the physical tests
(organoleptic, adhesion, homogeneity, pH, and viscosity) which
met the requirements at day 0. Subsequently, the optimized
formula was subjected to antibacterial activity against
P. acnes.
Antibacterial Activity Test
P. acnes were tested for antibacterial activity using agar
diusion method anaerobically as described by Jusuf et al.26
The materials and tools were sterilized before use. Blood base
agar was made by dissolving 10 grams of media in 250 mL of
distilled water. The media was then sterilized for 15 minutes
at 121°C and poured into a petri dish once the temperature
had dropped to 40– 45°C. P. acne suspension in 5 mL was
subsequently added into the petri dish and the turbidity was
assessed using 0.5 McFarland. Following the inoculation of
P. acnes bacteria into the media, four wells were prepared
in each petri dish using a sterile tip. The samples were
erythromycin, citronella-scented oil, sodium benzoate,
optimum cream formula, and distilled water. The plates were
then incubated at 37°C for 24 hours. Replication was carried
out three times. Observations were conducted by measuring
the inhibition zone which was formed.

Formulation of Citronella Oil Cream for Acne Treatment
IJDDT, Volume 13 Issue 1, January - March 2023 Page 420
Data Analysis
Data on organoleptic test results, pH, cream type, emulsion
height, specific gravity and homogeneity were analyzed
descriptively. While the results of the adhesion and viscosity
test at day 0, and the antibacterial test were analyzed using
one-way anova method with a 95% condence level (SPSS
version 26).
RESULTS AND DISCUSSION
Citronella oil contains citronellal, citronellol, and geraniol
compounds that act as antibacterial agents.9 -11 Nevertheless,
citronella oil cannot be used directly since it might irritate.13
As a result, in this study citronella oil was formulated in cream
dosage form.
Cream is a suitable dose form for hydrophobic compounds
since it comprises water and oil phase.15 The requir e d oil HL B
must rst be optimized to identify the surfactant accounts for
the most stable citronella oil cream. In this study, we used
the combination of span 60 and tween 60 as emulsiers. The
creaming index determines the concentration of tween and
span is close to 1, where the change of emulsion height was
minimum.27 The surfactant concentrations for tween 60 was
3.4% and for span 60 was 1.6% since they had the highest
creaming index, 0.93 (data not shown). The ratio value was
selected since less creaming will form in the emulsion as the
ratio closes to 1.
Furthermore, optimization of cetyl alcohol as a viscosity
enhancer was performed at five different formulas with
cetyl alcohol concentrations ranging from 2 to 10%. The
ve formulas underwent emulsion type, pH, homogeneity,
adhesion, specic gravity, viscosity, and stability test.
Figure 1 displays the organoleptic eects of citronella oil
cream. The citronella oil cream formed in this study had a white
and semi-solid cream with a distinctive odor. No organoleptic
changes or separation were observed during the 28-day storage
at room temperature 25°C and freeze-thaw test in all of the
ve formulas. The ndings hence indicated that ve cream
formulas were stable throughout the storage.
Cream is homogeneous when no obvious coarse granules
are visible.21 The homogeneity test revealed that the ve
formulations were all homogeneous. Phase separation was not
observed during 28-day storage at room temperature (25°C)
and freeze-thaw at room temperature.
The five cream formulas can be diluted with water,
indicating that they are an o/w (oil in water) emulsion type.
Cream with O/W emulsion-type is favored since it is water-
rinsable and does not leave a greasy feeling.28
Figure 2 shows the result of the pH test. The physiological
pH value of the skin ranges from 4.5-6.5.29 The pH test
revealed that formulas 3, 4, and 5 fullled the skin pH, but
formulas 1 and 2 had higher pH values 6.6–6.8. The pH was
increased at lower cetyl alcohol concentration. This result
agrees with Handayani et al., which found that the pH value
will decrease along with the increase in cetyl alcohol content.30
The ve formu l as pr ove d stab le be c ause stora ge und e r amb ient
Figure 3: Viscosity results of citronella oil cream at room temperature
storage (A) and freeze-thaw (B) stability test. Citronella oil with cetyl
alcohol 2% (F1); 4% (F2); 6% (F3); 8% (F4); 10% (F5).
Figure 1: Organoleptic results of citronella oil cream. Citronella oil
with cetyl alcohol 2% (F1); 4% (F2); 6% (F3); 8% (F4); 10% (F5).
Figure 2: pH results of citronella oil cream at room temperature storage
(A) and freeze-thaw (B) stability test. Citronella oil with cetyl alcohol
2% (F1); 4% (F2); 6% (F3); 8% (F4); 10% (F5).

Formulation of Citronella Oil Cream for Acne Treatment
IJDDT, Volume 13 Issue 1, January - March 2023 Page 421
conditions and freeze-thaw cycles revealed no signicant pH
changes.
Figure 3 displays the viscosity test results. A good cream
vis cosity was bet we e n 2,00 0 an d 50,000 mPa s.31 The viscosity
values of the ve formulas were in the ranges of 2798–6364 mPas
for formulas 2, 3, 4, and 5, whereas formula 1 did not meet good
cream requirements since the values were 1011–1287 mPas.
The viscosity of cream increases signicantly with the increase
of cetyl alcohol concentration (from F1 to F5). This result
agrees with Kang et al. and is consistent with the nature of
cetyl alcohol as a viscosity enhancer.32
The viscosity result is in agreement with the cream-specic
gravity result, where the increase of cetyl alcohol concentration
increased the specic gravity. Formula 1 has the lowest specic
gravity (0.973 + 0.009 g/mL), while formulas 2, 3, 4, and 5
increased accordingly (1.006 + 0.004, 1.031 + 0.009, 1.055 +
0.006, and 1.082 + 0.007 g/mL, respectively). Previous study
found that a good cream specic gravity ranging from 0.95 to
1.05 g/mL,31 hence the formulas 1, 2, 3, and 4 met the criteria.
Statistics analysis reveals that there are no notable specic
gravity changes upon room temperature storage.
The subsequent test is an adhesion test. The adhesive
result of ve formulations increases signicantly ( p < 0.05)
with the increase of cetyl alcohol, as shown in Figure 4. This
result agrees with Kulawik-Pióro et al. which mentioned
that cohesiveness and index of viscosity were related to the
cream adhesiveness.33 The adhesive requirement for a cream
preparation is not less than 4 seconds, hence five cream
formulas fullled the acceptance criteria of good cream.34
The optimum preparation will be chosen for antibacterial
testing by analyzing the physical parameters during room
temperature storage and freeze-thaw. Formula 3 was chosen
for the antibacterial test out of all the formulas tested because
it produced the best test results, where formula 3 fulls all
test requirements for acceptable physical properties of cream
preparations.
For the antibacterial test, the average result of the inhibition
zone against P. ac nes for cream formula 3 was 9.35 mm,
citronella oil was 29.22 mm, and erythromycin was 27.33 mm.
The aquadest, sodium benzoate, and cream base did not
produce any inhibition zone, indicating that the components
did not aect the optimum cream formula inhibition zone and
that the formula 3 inhibition zone are proven to come from
the active substance.
Compared to citronella oil cream in optimum formula
(formula 3), the results of erythromycin and citronella oil
inhibitory zones were higher. This result is in accordance
with El-Gied et al.35 Cunha et al. mentioned that antimicrobial
agents (such as thymol) bind to tween and reduce their
antibacterial activity.36 The active component of essential oils
usually dissolved in hydrophobic bases, thereby preventing
their release due to their strong anity for the lipophilic
components (semi-solid formulation coecient partitions).37-38
Based on the results of the investigation, formula 3 with
a concentration of 6% cetyl alcohol is the optimum cream
formulation and has been shown to have antibacterial action
against the P. ac nes.
CONCLUSION
The optimum concentration of cetyl alcohol that produces the
best cream preparation is 6%. Citronella oil cream preparations
possess potential antibacterial activity against the acne-causing
bacteria, P. acnes.
ACKNOWLEDGEMENT
The author would like to thank Institut Technology Sumatera
for providing the laboratory equipment and instrument.
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