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The 5th International Conference on Pharmaceutical Nanotechnology / Nanomedicine 2021 | 84
FORMULATION AND EVALUATION OF CITRONELLA OIL IN ROLL-ON APPLICATION SYSTEM
Original Article
SAFIRA NAFISA1, SHIRLY KUMALA1, YATI SUMIYATI1*, WIWI WINARTI1, DIAH KARTIKA PRATAMI1, DESI
NADYA AULENA1, NOVI YANTIH1, AMELIA NURSAFITRI1
1Faculty of Pharmacy, Pancasila University, South Jakarta, DKI Jakarta, 12640, Indonesia
*Email: yati.sumiyati@univpancasila.ac.id
Received: 11 Dec 2021, Revised and Accepted: 21 Mar 2022
ABSTRACT
Objective: The purpose of this study to obtain the best formulation of roll-on Citronella oil that can be used for aromatherapy.
Methods: The citronella plant (Cymbopogon winterianus Jowitt) was processed with Steam-water distillation produced essential oil. Citronella oil
was tested for quality and formulated. The best formulation result is then tested for product quality.
Results: The result of yield of citronella oil was 0.6%. The result of citronella oil quality parameters is obtained in accordance with the literature.
The result of the hedonic test, formula 2 was chosen to be the best formula. The result of an average transferred volume was 10.0 ml. The result of
the total plate number is<`10 cfu/ml. The result of the identification of Staphylococcus aureus and Pseudomonas aeruginosa was negative. The result
of the stability test was stable during storage.
Conclusion: The results showed that formula 2 was the best and has the potential to be circulated in large quantities
Keywords: Citronella oil, Roll-on, Aromatherapy, Cymbopogon winterianus Jowitt
© 2022 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/)
DOI: https://dx.doi.org/10.22159/ijap.2022.v14s3.17 Journal homepage: https://innovareacademics.in/journals/index.php/ijap
INTRODUCTION
Citronella oil is an essential oil obtained from distillation of the
leaves of Lemongrass Cymbopogon winterianus and Cymbopogon
nardus. Citronella oil contains the main component in the form of
citronellal and geraniol, which can give a distinctive odor such as the
aroma of grass that will be liked by the public. Cymbopogon
winterianus produced higher levels of citronellal and geraniol than
Cymbopogon nardus. Citronella oil is one of the essential oil
commodities with high demand for export by Indonesia and it still
needs to be developed to increase the selling value [1]. One of the
efforts to add selling value of citronella oil in Indonesia is can be
made into aromatherapy products because in the 21st century,
aromatherapy has received a lot of attention [2, 3].
Aromatherapy comes from the word “aroma” which means the smell
of fragrance and “therapy,” which means treatment with the aim of
helping provide a feeling of freshness and calm [4, 5]. An
aromatherapy roll-on made from Citronella oil mixed with base oil
The right one will provide a mild warm sensation and the aroma of
Citronella oil can help relieve headaches and relieve stress because it
has a soft aroma that can reduce mental tension [6]. Based on
research, the result of the distillation of citronella oil can function as a
sedative in male mice [7]. Research on the formulation and stability of
roll-on aromatherapy based on Citronella oil is still limited; therefore,
this research will conduct formulation and test the stability of roll-on
aromatherapy preparations containing Citronella oil.
MATERIALS AND METHODS
Plant material
The main ingredients used in this study were Lemongrass
(citronella) leaves (Cymbopogon winterianus Jowitt Ex Bor) obtained
from Bukit Waruwangi, Serang, Banten, Indonesia.
Chemical ingredients
Menthol, Camphora, BHT (butylated hydroxyanisole),
Propyleneglycol, Lemon oil and Virgin coconut oil were purchased
from Faculty of Pharmacy, Pancasila University.
Plant determination
Plant determination was carried out at the Herbarium Depokensis
(DEB), Departement of Biology, Faculty of Science and Mathematics,
Universitas Indonesia, Depok, West Java (No.701/UN2.
F3.11/PDP.02.00/2021). The purpose of determination is to get to
the truth and clear identity of the plant researched and avoid error
in the main research material collection [8].
The distillation of citronella oil
Lemongrass leaves (Citronella) distillation was carried out using the
steam-water distillation method to obtain Citronella oil [9].
Measurement of citronella oil yield
The purpose of measurement of citronella yield was to get how
many grams of simplicia powder to produce 1 gram of extract [10].
The yield of the extract was calculated with the formula Eq. 1:
% Yield of extract = Extract weight obtained (g)
sampel weight (g)x100% …… (1)
Determination of citronella oil quality parameters
Determination of quality requirements for Citronella oil refers to SNI
06-3953-1995 [11]. The parameters of color testing was carried out
organoleptically. The density test was carried out using a
pycnometer. The refractive index test was carried out using a
refractometer. The total geraniol and citronellal test was carried out
using gas-liquid chromatography. The solubility in ethanol 80%
testing was carried out using the synthesis method and foreign
substances such as fatty oil.
Citronella oil roll on aromatherapy formula
The roll-on aromatherapy formula was shown in table 1.
Making aromatherapy roll-on citronella oil
Prepare the necessary tools and materials. Calibrated roll-on
container ad 10 ml. The menthol and camphor were weighed in
different glasses and mixed, then set aside to form M1 (Mass 1). BHT
was weighed and dissolved with some base oil, then set aside to
become M2 (Mass 2). Put M1 and M2 into the beaker glass.
Propylene glycol was measured put into a beaker glass and stirred
homogeneously. In Formula 3, lemon oil was measured and put into
a beaker glass and stirred homogeneously. Put the mixture into the
roll-on container. Added the remaining base oil ad calibration mark.
Shaken ad homogeneously and equipped with a roll-on and labeled
on the primary packaging. Put in secondary packaging.
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ISSN- 0975-7058 Vol 14, Special Issue 3, 2022
S. Nafisa et al.
Int J App Pharm, Vol 14, Special Issue 3, 2022, 84-87
The 5th International Conference on Pharmaceutical Nanotechnology / Nanomedicine 2021 | 85
Table 1: Formula design
No Ingredients Formula (%)
1 2 3
1.
Citronella oil
10
5
5
2.
Menthol
30
30
30
3.
Camphor
5
5
5
4.
BHT
0.2
0.2
0.2
5.
Propylene glycol
15
15
15
6.
Lemon oil
-
-
5
7.
Virgin coconut oil
39.8
44.8
39.8
Roll-on citronella oil quality evaluation
Product quality evaluation is carried out referring to the National
Agency of Drug and Food Control of Indonesia Reg. PERKABPOM No.
32 of 2019 includes organoleptic tests, transferred volumes, and
microbial contamination. Stability tests, preparations, primary
irritation tests, and hedonic tests [12].
RESULTS AND DISCUSSION
Plant determination
The results of plant determinations carried out at the Faculty of
Mathematics and Natural Sciences, University Indonesia, showed
that the plants used were true surrender plants (Cymbopogon
winterianus Jowitt Ex Bor) with the Poaceae family. Results the
determination is based on the Book Flora of Java Vol. III [13].
Measurement of citronella oil yield
The yield of the steam distillation of Cymbopogon winterianus was
shown in table 2.
Table 2: Result citronela oil yield
Sample weight (g)
Extract weight obtained (g)
Yield (%)
680.000
4.100
0.6
After the extraction process is complete, the result of measurement
Citronella is 0.6%. The yield of extraction in this study lower than overall
oil yield obtained by Singh of Citronella essential oil was 0.79% [14]. The
methods of distillation process influenced citronella oil yield. The
modern extractor machine showed best-distilled efficiency of oil content
than the traditional alcohol boiler that shown lowest on oil content [15].
Determination of Citronella oil quality parameters
The quality parameter of citronella oil in this study was shown in table 3.
Based on the results obtained are following SNI 06-3953-1995, This
means that the resulting product can be used as raw material for
manufacturing roll on citronella oil as aromatherapy. Citronella oil
that meet the requirement can be used on the different product as
aromatherapy, insect repellent, medical, food product, and cosmetic
[15]. The best quality of citronella oil was had various benefits and
offered potential essential options which could be further applied to
many applications
Hedonic test
Test of interest in the final results of roll-on aromatherapy preparations
with an assessment using numerical parameters based on warmth and
aroma. The rating scale consists of 5, namely: very like, like, neutral,
don't like, and don't like it. The number of panellists consists of 30
people. The data obtained is processed using the software SPSS®25. The
results of the hedonic test are presented in table 4.
Table 3: Certificate of analysis sheet Citronella oil
Parameter Specification Results
Appearance
Clear, sometimes slightly opalescent, mobile liquid
According to the standard
Colour
Colourless to pale yellow
According to the standard
Fragrant
Fresh, citrusy, sweet, slightly spicy
According to the standard
Density (25 °C)
0.8750-0.8930
0.8828+0.02
Refractive index (20 °C)
1.460–1.474
1.472+0.12
Optical Rotation
-7.00 °–0.00 °
-0.35+0.01 °
Citronella content
Minimum 35%
32.22+1.23%
Geraniol Content
17-30%
29.25+1.15%
Solubility
Soluble in alcohol and oil. Not soluble in water
Conform to the standard
Fat oil
Negative
Negative
Data was given in mean+SD, n=3
Table 4: Result hedonic test
Warmth
Scent
Duncan
a,b
Duncan
a,b
Subset
Sample
N
1
2
Sample
N
1
2
Formulation 1
30
3.83
Formulation 3
30
3.77
Formulation 2
30
3.97
Formulation 2
30
3.77
Formulation 3
30
4.33
Formulation 1
30
4.23
Sig.
.375
1.000
Sig.
1.000
1.000
Based on the hedonic test, it was known that each roll-on formula,
namely, 10% citronella oil (Formula 1), 5% citronella oil (Formula 2),
and a mixture of 5% citronella oil and 5% lemon (Formula 3) can be
concluded that formula 2 is more dominant and much in demand by
many people in terms of warmth and aroma. Therefore, formula 2
was chosen to be the best formula.
Transferred volume test
Based on the results of the tests carried out, the average transferred
volume was 10.0 ml from 10 repetitions. The results obtained
requirements determined by the regulatory standards of the drug
and food regulatory agency number 32 of 2019. The substandard of
S. Nafisa et al.
Int J App Pharm, Vol 14, Special Issue 3, 2022, 84-87
The 5th International Conference on Pharmaceutical Nanotechnology / Nanomedicine 2021 | 86
volume is caused by several factors such as the method, labour, and
machine, while the most potential factor is the volume conformity to
reduce the number of defect products.
Microbial contamination testing
The result of the total plate count was shown in table 5.
Table 5: Result total plate count
Sample
Dilution
Result (cfu/ml)
10-1 10-2 10-3 10-4 10-5
Roll on-1
0
0
0
0
0
<10 cfu/ml
Roll on-2
0
0
0
0
0
In the total plate count test, 5 dilutions were carried out in duplicate
on the sample. Based on the results of testing the total plate number
in the roll-on Citronella oil aromatherapy preparation, there were no
growing colonies (<10 cfu/ml). The Total Plate count required for
external medicinal liquids according to the regulation of the drug
and food regulatory agency number 32 of 2019 was 107 cfu/ml. So, it
can be concluded that the tested Citronella oil roll on aromatherapy
preparations met the specified requirements and was
microbiologically safe for use by the public.
The identification of Staphylococcus aureus contamination was
shown in table 6.
Table 6: Result identification of Staphylococcus aureus
Identification of microbial contamination
Media
TSB MSA
Roll on-1
Turbid
-
Roll on-2
Turbid
-
Positive control
Turbid
fluorescence green colony
Negative control
Clear
-
Description: TSB: Tryptic Soy Broth; MSA: Mannitol Salt Agar
In the Staphylococcus aureus identification test, Tryptic Soy Broth
(TSB) media was used as an enrichment medium and if the results
were positive, then it would be carried out on Mannitol Salt Agar
(MSA) media. Based on the results obtained on Tryptic Soy Broth
(TSB) media is positive, this is indicated by turbidity and deposits
at the bottom of the tube, so it is necessary to continue testing
using selective media, namely Mannitol Salt Agar (MSA) by
scratching. On Mannitol Salt Agar (MSA) media, negative results
were obtained because it did not show the formation of yellow or
white colonies surrounded by a white zone. Staphylococcus aureus
which is required for external drug fluids according to the
regulation of the drug and food regulatory agency number 32 of
2019 is negative/ml.
The identification of Pseudomonas aeruginosa contamination was
shown in table 7.
Table 6: Result identification of Pseudomonas aeruginosa identification of microbial contamination
Media
TSB
CETA
Roll on-1
Turbid
-
Roll on-2
Turbid
-
Positive control
Turbid
Yellow colony
Negative control
Clear
-
Description: TSB: Tryptic Soy Broth; CETA: Cetrimide Agar
In testing the identification of Pseudomonas aeruginosa, Tryptic Soy
Broth (TSB) media was used as an enrichment medium and if the results
were positive, then it would be carried out on Cetrimide agar media.
Based on the results obtained on Tryptic Soy Broth (TSB) media is
positive, this is indicated by turbidity and deposits at the bottom of the
tube, so it is necessary to continue testing using selective media, namely
Cetrimide by scratching. On Cetrimide agar media, negative results were
obtained because it did not show the formation of fluorescent green
colored colonies. Pseudomonas aeruginosa required for external
medicinal fluids according to the regulation of the drug and food
regulatory agency number 32 of 2019 was negative/ml.
Stability test
The stability test of citronella oil was shown in table 7 below.
Table 7: Stability test of citronella oil
Time
Description
pH
Room temperature (25 °C) Temperature 40 °C
Week 0
Colorless, clear liquid, characteristic aromatic odor
6.2
-
1st week
Colorless, clear liquid, characteristic aromatic odor
6.32
6.33
2nd week
Colorless, clear liquid, characteristic aromatic odor
6.75
6.75
3rd week
Colorless, clear liquid, characteristic aromatic odor
6.21
6.41
4th week
Colorless, clear liquid, characteristic aromatic odor
6.44
6.40
The pH of the preparation for topical preparations must have the
same pH range as the skin, namely 4.5-7.0. If the pH of the
preparation is too low or too acidic, it will irritate the skin so that a
burning sensation will appear, while if it is too alkaline, it will give a
dry and itchy feeling. Based on the results, it can be concluded that
the pH obtained is still within the desired pH range and does not
S. Nafisa et al.
Int J App Pharm, Vol 14, Special Issue 3, 2022, 84-87
The 5th International Conference on Pharmaceutical Nanotechnology / Nanomedicine 2021 | 87
have the potential to irritate. Besides that, there is no change in
shape and aroma during 1 mo of storage. This shows that VCO is a
good carrier oil capable of binding the mixed essential oils.
CONCLUSION
The results showed that formula 2 is the best and has the potential
to be circulated in large quantities.
ACKNOWLEDGMENT
This article is part of the study independent of MBKM Pancasila
University. We acknowledge the financial support from the Ministry
of Education and Culture and Higher Education, Republic of
Indonesia, through Matching Fund Grant 2021.
FUNDING
This work was supported by the Ministry of Education and Culture
Republic of Indonesia through Matching Fund Grant (MoU No:
4026/E3/PKS.09/KL/2021 and No: 5832/R. UP/PKS/IX/2021).
AUTHORS CONTRIBUTIONS
All the authors have contributed equally.
CONFLICT OF INTERESTS
The author declares no conflict of interest.
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