Alexandria Engineering Journal (2023) 70, 569–583

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Alexandria University

Alexandria Engineering Journal

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ORIGINAL ARTICLE

Extraction of citronella oil from lemongrass

(Cymbopogon winterianus) by sequential ultrasonic
and microwave-assisted hydro-distillation
Maya Sarah a,b,*, Dwiky Ardiansyah a,b, Erni Misran a,b, Isti Madinah a,b

a
Chemical Engineering Department, Universitas Sumatera Utara, Indonesia
b
Jl. Almamater Kampus USU, Padang Bulan, Medan 20155, Indonesia

Received 28 January 2022; revised 2 July 2022; accepted 5 March 2023

KEYWORDS Abstract The Ultrasonic and Microwave-Assisted Hydro-Distillation (US-MAHD) method is a

Extraction; process combination of ultrasonic-assisted extraction (UAE) and microwave-assisted hydro-
Essential oil; distillation (MAHD) carried out sequentially. This method aims to improve the UAE and MAHD
Lemongrass; methods in extracting citronella oil. This study evaluates the performance of US-MAHD in the
Microwave; extraction of citronella oil from the citronella plant. Extraction is conducted for 90 min at various
Ultrasonic solvent-plant ratios (v/m) (10:1, 12:1, 14:1), ultrasonic bath temperature (30 °C, 40 °C, 50 °C), and
power of microwave oven (150 W, 300 W, 450 W). US-MAHD yields a maximum yield of citronella
oil of 1.82 mg/g when extracting lemongrass plant with a combination of ultrasonic bath tempera-
ture of 30 °C, a solvent-plant ratio of 10:1, microwave power of 300 W, and time of 90 min. All
quality parameters of the oil produced have met ISO 3848:1976 standard. Extraction of citronella
oil using the US-MAHD method produces higher yields than the UAE and MAHD methods under
the same operating conditions. The yields of citronella oil from the US-MAHD, UAE, and MAHD
methods are 1.82, 0.92, and 1.48 mg/g, respectively. SEM analysis of residual lemongrass plant
shows more cell wall damage which indicates more oil release from the plant matrix.
Ó 2023 THE AUTHORS. Published by Elsevier BV on behalf of Faculty of Engineering, Alexandria
University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/
licenses/by-nc-nd/4.0/).

1. Introduction roots, leaves, bark, stems, fruit, seeds, or flowers of aromatic

plants results in an essential oil [1]. The essential oil is sec-
Indonesia has many exotic aromatic plants containing complex ondary metabolites product of plants that has specific fragrant
volatile compounds such as lemongrass, lime orange, cinna- [2]. Essential oil, therefore, becomes a key ingredient to pro-
mon, Javanese frankincense, sandalwood, etc. Extraction of duce perfume, soap, cosmetics, etc. Lemongrass plant with
an essential oil content of 1–2% (dry basis) [3] is available from
* Corresponding author. abundant sources in Sumatera Utara Province, Indonesia, but
E-mail address: maya3@usu.ac.id (M. Sarah).
their market and utilization are low [4]. The essential oil of the
Peer review under responsibility of Faculty of Engineering, Alexandria
lemongrass plant known as citronella oil is characterized by a
University. strong lemon fragrance due to the citral compound in the oil
https://doi.org/10.1016/j.aej.2023.03.019
1110-0168 Ó 2023 THE AUTHORS. Published by Elsevier BV on behalf of Faculty of Engineering, Alexandria University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
570
[5]. Commercial citronella oil comprises as Ceylon citronella
and Java type citronella. The Ceylon citronella oil is extracted
from Cymbopogon nardus while the Java type is from Cymbo-
pogon winterianus [6].
In industry, citronella oil is one of the ingredients used to
produce ionone, vitamin A, and carotene [3]. The aroma of
the oil is widely used in soaps, detergents, and perfumes. In
addition, citronella oil contains several bioactive compounds
that are medicinal [5], antioxidants, anti-microbial, and anti-
fungal [3]. Citronella oil also functions as an insect repellent
because of its high efficacy and low toxicity, this use has been
registered with the US EPA (Environmental Protection
Agency) [7]. Although citronella oil is widely used in the phar-
maceutical, cosmetic, and food industries, its selling price is
still low, so it is needed to increase its economic value by iso-
lating its active components such as citronellal, citronellol, and
geraniol [8].
Extraction of essential oils from exotic aromatic plants can
be carried out by several methods such as conventional steam
distillation and hydro-distillation solvent extraction [9], inno-
vative methods such as sonoprocess-assisted solvent extraction
[1011], ultrasonic-assisted extraction (UAE) and microwave-
assisted extraction (MAE) [1112]. Comparisons between tradi-
tional methods and innovative methods in the essential oil
extraction process have been reported by several authors else-
where. Hydro-distillation as a traditional method is a common
method for extracting essential oils, resulting in low extraction
yields with long processing times [13]. The hydro-distillation
method has been improved by using a microwave to aid extrac-
tion. Ranitha et al. [3] reported that at the same time the
extraction process, microwave-assisted hydro-distillation
(MAHD) resulted in a higher yield of citronella oil compared
to hydro-distillation and the content of citronella oil obtained
from both methods was almost the same. Hamdi et al. [14]
reported that the extraction process of Eucalyptus salubris
essential oil with MAE resulted in a higher yield of UAE
and hydro-distillation, while the UAE produced more
monoterpene hydrocarbon compounds.
Hamzah et al. [15] extracted citronella using three methods,
namely ohmic-heated hydro-distillation, hydro-distillation,
and steam distillation. Ohmic-heated hydro-distillation was
considered feasible as an alternative method of extracting
citronella oil because the SEM test of citronella grass showed
that ohmic heating caused the formation of transient pores in
the cell membrane, although the cell wall was still rigid. Ghaz-
anfari et al. [16] reported that essential oil from coriander seeds
extracted with MAHD showed better antimicrobial activity,
higher phenols yield, and antimicrobial activity than the
hydro-distillation method. Nurul et al. [17] extracted emprit
ginger oil using four methods, namely microwave ultrasonic
steam diffusion (MUSDf), steam diffusion (SDf), microwave
extraction (ME), and microwave steam diffusion (MSDf),
the best results were obtained using the MUSDf method with
a yield of 0.952%.
Dranca et al. [18] compared conventional extraction and
microwave-assisted extraction to obtain pectin from Malus
Domestica ’Falticeni’ pomace, both methods obtained the
same pectin yield but microwave extraction required a shorter
time, galacturonic acid content and the esterification rate of
pectin obtained was similar in both methods. Microwave-
extracted pectin is possible for the manufacture of films having
low oxygen permeability. Alara, et al. [19] reported the extrac-
M. Sarah et al.
tion of oil from Carica papaya leaves, the results of this study
showed that irradiation time, microwave power, and solvent
volume significantly changed the oil yield of C. papaya. The
best yield obtained was 3.25 ± 0.25% v/w at an irradiation
time of 120 min, microwave power of 500 W, a solvent volume
of 200 mL, and a temperature of 100 °C.
Silva et al. [20] conducted a study on the extraction of pas-
sion fruit seeds (Passiflora edulis Sims) to obtain residual seed
cake from oil extraction containing piceatannol, the study was
carried out using microwave-assisted extraction method and
conventional Soxhlet extraction method. Microwave extrac-
tion yielded a fine cocoa powder with 27.17 ± 0.9 g piceatan-
nol per mg extract at 87 °C, with 70% ethanol, for 30 min each
cycle, whereas Soxhlet extraction yielded a dark extract con-
taining 13.03 ± 0.4 g/mg for 120 min. This proved that micro-
wave extraction is a promising alternative for extracting
passion fruit seed and adding to the residual value of passion
fruit by providing a faster extraction, more color friendly,
and yielding higher piceatannol compared to a conventional
method. Hu et al. [21] found that the essential oil by micro-
wave extraction had better quality than Soxhlet extraction,
especially with low acid and peroxide values.
Yingngam et al. [22] reported for the first time the extrac-
tion and identification of volatile compounds in essential oils
from Shorea roxburghii flowers. Essential oil isolation using
solvent-free microwave extraction method, the extracted essen-
tial oil showed better overall parameters than traditional
hydro-distillation methods, including shorter isolation time,
higher odor quality, and environmental compatibility. Leema
et al. [23] conducted a study on microwave-assisted extraction
to extract lutein from Chlorella sorokiniana (NIOT-2). X-ray
diffraction (XRD) analysis of the microwave-treated biomass
(83.85%) showed a much higher crystallinity index than the
untreated sample (17.28%). Microwave pretreatment is con-
sidered suitable for the extraction of lutein from marine
microalgae in consideration of the speed of coalescence, homo-
geneous heating, less energy intensity, and high extraction
yield.
Both UAE and MAE have been shown to improve over-
all extraction performance in terms of yield, oil quality, and
extraction time [24]. In the extraction process using UAE,
ultrasonic waves will produce cavitation bubbles which
can cause cell wall damage so that compounds in cells
can be extracted [25]. While MAE uses microwave energy
which causes polar molecules in the plant to reorient fol-
lowing the direction of the magnetic field movement contin-
uously, causing heat from friction between molecules and
heat coming out of the plant sample accompanied by evap-
oration of essential oils from plant cells. This process
increases the effectiveness of the extraction process includ-
ing a shorter extraction time [26]. In addition to a shorter
time, the microwave method offers an important advantage
over traditional methods, namely producing a more valu-
able essential oil with a high amount of oxygenated com-
pounds [27].
Rodsamran and Sothornvit [28] reported extraction of phe-
nolic compounds from lime peel waste using ultrasonic-
assisted and microwave-assisted extraction showed that the
UAE method was more effective for extracting total phenolics
(54.4 mg GAE/g) with high antioxidant activity and 33% time
saving compared to MAE. One of the other advantages of
applying ultrasound in the extraction of essential oils from
Extraction of citronella oil from lemongrass (Cymbopogon winterianus) by sequential ultrasonic
medicinal plants is the use of less solvent compared to conven-
tional methods such as mechanical and Soxhlet extraction [29].
Sharifzadeh et al. [30] studied the essential oil of Lavandula
coronopifolia Poir using a sequential ultrasonic-microwave
technique. The highest yield was obtained at 1.15%, the value
was 82% higher than the yield of the basic method. This
method produced a much higher number of active ingredient
compounds than the microwave method, and more antioxi-
dant properties. The active ingredients obtained from this
extraction method had the highest antimicrobial properties.
In addition, it is more environmentally friendly than the
microwave-assisted extraction method.
Ultrasonic also shows tremendous potential in synthesizing
nanomaterials, particularly nano biomaterials for biomedical
applications. The ultrasonic synthesis produces nano biomate-
rials with better properties and performance than conventional
synthesis methods and eliminates the need for harsh and
expensive chemicals [31]. Ngamkhae et al. [32] extracted the
Kleeb Bua Daeng formula with ultrasonic and microwave
extraction methods separately. Extracts from the microwave
method contained higher phenols, flavonoids, carotenoids,
and anthocyanins than extracts from the ultrasonic method.
The microwave method is considered more efficient in extract-
ing the formula. A comparative study of ultrasonic and micro-
wave extraction was also reported by Sharma and Dash [33] to
extract phytocompounds from black Jamun pulp. This method
will make it possible to extract polyphenols and anthocyanins
from black Jamun pulp for nutraceuticals, pharmaceuticals,
and functional food-based industries.
Yang et al. [34] have successfully developed microwave-
induced hydro-distillation and simultaneous extraction in a
rotary state to produce essential oil, rosmarinic acid, and
polysaccharides from Perilla frutescens. This technique can
be an alternative to improve the extraction of essential oils,
polysaccharides, and other target analytes from P. frutescens
or other plant materials. Costa et al. [35] have developed a
greener microwave-assisted extraction method by modifying
it using a natural deep eutectic solvent that is applied for the
preparation of medicinal plant samples. The optimization of
the combined microwave method and the use of environmen-
tally friendly methods resulted in excellent analytical
parameters.
Wan et al. [36] simultaneously extracted essential oil and
flavonoids from Baeckea frutescens by enzyme pre-treatment
method combined with ultrasonic microwave-assisted surfac-
tant. The concentration of flavonoid extract obtained was
9.7 times higher than the original extract. The essential oil
obtained was confirmed to have potential bioactivity. These
results indicated this method can be used as a promising,
and environmentally friendly alternative to extracting the
active constituents.
However, in the extraction process, UAE and MAE meth-
ods have disadvantages [3738]. UAE has the problem of atten-
uation effect [39] while in the extraction process using MAE,
inhomogeneous heating can occur [40]. Because of these short-
comings, Yu et al. [37] evaluated the extraction process of total
flavonoids using sequencing of UAE and MAE (US-MAE)
from Osmanthus fragrans Lour. flowers. Yu et al. [37] reported
that US-MAE resulted in a higher yield of total flavonoids
than UAE and MAE.
To date, no study reported extraction of essential oil using a
sequential combination of UAE and MAHD (US-MAHD),
571
especially for the extraction of citronella oil. Therefore, this
study aims to extract the essential oil of lemongrass leaves
(Cymbopogon winterianus) using US-MAHD. The effects of
solvent to plant ratio, ultrasonic temperature, and microwave
power on yield are evaluated. To compare the effectiveness of
the US-MAHD method, extraction of lemongrass leaves with
UAE and MAHD is carried out using the temperature, fre-
quency, and power of US-MAHD which results in the highest
yield. The performance of US-MAHD will be compared with
UAE and MAHD based on the yield and the constituents of
citronella oil. To maximize the yield of citronella oil from
the lemongrass plant, US-MAHD extraction of lemongrass
stems is also investigated. Overall, this study is expected to
increase the quality and quantity of citronella oil production
in Indonesia, especially in Sumatera Utara Province which is
still done traditionally using the steam distillation process that
is considered ineffective and very expensive due to high energy
requirements.
2. Materials and methods
2.1. Materials
Materials in this study comprise lemongrass (Cymbopogon win-
terianus) taken from Deli Serdang, Sumatera Utara Province,
Indonesia, distilled water as solvent from the local chemical
supplier (PT. Rudang, Medan), and ethanol 80% for alcohol
solubility analysis supplied by PT. Merck Tbk. The extraction
process was carried out in an ultrasonic bath (Elmasonic S 300
(H)) for the UAE method and a microwave oven (Microwave
Sharp R-21D0(S)IN) for the MAHD method. The ultrasonic
bath is equipped with tank internal dimensions of 505 x300x
200 mm that operated at an ultrasonic frequency of 37 kHz.
The microwave oven was modified with the hydro-distillation
apparatus. The dimensions of the microwave oven cavity were
485
400
292 mm with 800 W of power consumption and a
229–240 Volt power source. This experiment was carried out at
the Laboratory of Organic Chemistry, Chemical Engineering
Department, Universitas Sumatera Utara, Medan, Indonesia.
2.2. Methods
The scheme of citronella oil extraction from lemongrass plants
by sequential US-MAHD method in this study is illustrated in
Fig. 1. Before US-MAHD, pre-treatment is conducted to pre-
pare the material to obtain the best result. The sequential US-
MAHD begins with the extraction by ultrasonic wave followed
by microwave irradiation assisted by hydro-distillation. The
extracted oil was separated from the solvent and dried to
remove all the water before a further investigation of oil qual-
ity (color, constituents, density, refractive index, and solubility
in ethanol).
2.2.1. Pretreatment of lemongrass
Fresh lemongrass plants collected from the plantation were cut
for the leaves and the stem. The leaves were dried at room tem-
perature without direct exposure to sunlight for a week. The
drying aims to reduce water content in the plant to increase
extraction yield [3]. The dried plants were cut with a dimension
of 1x1 cm and then ground using a grinder (Miyako
572 M. Sarah et al.

Fig. 1 Scheme of citronella oil extraction from lemongrass plant by sequential US-MAHD method in this study.

BL-101PL) with a power of 300 W for one minute. Samples
that have been reduced in size will facilitate the oil extraction
process with US-MAHD to obtain a high yield [41].
2.2.2. The sequential US-MAHD
In this study, the main ingredient was citronella leaves because
the fragrance was more dominant, while the stem was used as a
comparison. Experimental devices to perform sequential US-
MAHD are shown in Fig. 2. UAE and MAHD used to com-
pare US-MAHD performance are shown in Fig. 2(a) and 2(b)
respectively. About 100 g of the treated lemongrass leaves were
placed into the flask. The distilled water was added at various
ratios of solvent to plant material (v/m) (10:1; 12:1; 14:1
respectively). The flask was transferred into the ultrasonic bath
to extract the sample using the UAE method. The process was
carried out for 25 min at various ultrasonic bath temperatures

Fig. 2 Experimental apparatus for US-MAHD in this study (a) UAE and (b) MAHD: (1) transducer, (2) water, (3) ultrasonic bath, (4)
flask, (5) condenser, (6) microwave, (7) flask, (8) timer and power regulator, (9) condenser, (10) Liebig condenser, (11) separating funnel,
and (12) distillate bottle.
Extraction of citronella oil from lemongrass (Cymbopogon winterianus) by sequential ultrasonic
(30 °C; 40 °C; 50 °C respectively). After that, the flask was
placed into a microwave oven that have been connected to
hydro-distillation apparatus for further extraction process
using the MAHD method for 65 min. The microwave power
was varied at 150 W, 300 W, and 450 W. The procedure was
repeated for various combinations of solvent to plant material
ratio, the temperature of the ultrasonic bath, and microwave
power, each experimental run was conducted in triplicate.
Oil was separated from the distillate by using a separatory fun-
nel, followed by drying the oil over anhydrous sodium sulfate
to ensure there is no moisture content within the citronella oil,
and the yield obtained was calculated using Eq (1). As a com-
parison, extraction of citronella oil from the lemongrass stems
was then carried out using the best combination of tempera-
ture, frequency, and power of US-MAHD yielded the highest
oil from the lemongrass leaves.
ammount of essential oil obtained ðmgÞ
Yield ðmg=gÞ ¼
ammount of raw material used ðgÞ
ð1Þ
2.2.3. The UAE method
Distilled water as solvent was added into a flask containing
100 g of lemongrass leaves concerning solvent to plant ratio
obtained from the highest yield in the extraction of similar
material by US-MAHD. The sample was then extracted using
an ultrasonic bath with a frequency of 37 kHz for 90 min at the
ultrasonic temperature of US-MAHD resulting in the highest
yield. The experimental run was conducted in triplicate. The
separation of distillate was then carried out using a separatory
funnel and oil was dried further over anhydrous sodium sul-
fate. The experimental apparatus for the UAE method in this
study is shown in Fig. 2 (a).
2.2.4. The MAHD method
The microwave oven modified with hydro-distillation appara-
tus was used to extract 100 g of lemongrass leaves. The extrac-
tion process was carried out for 90 min using the ratio of
solvent to plant material resulting in the highest yield in the
extraction of citronella oil from lemongrass leaves by US-
MAHD. The experimental run was conducted in triplicate.
Microwave power was regulated in accordance power of US-
MAHD that yielded the highest yield. The distillate was col-
lected, and oil was separated using a separatory funnel and
dried further over anhydrous sodium sulfate to remove all
the solvent. The experimental apparatus for the MAHD
method in this study is shown in Fig. 2(b).
2.2.5. Determination of citronella oil properties
The Standard of density, refractive index, and oil solubility in
ethanol in this study is referred to as ISO 3848:1976. Determi-
nation of citronella oil’s density (20/20 °C), refractive index
(20 °C), and solubility in 80% (v/v) ethanol (20 °C) were car-
ried out following procedure of ISO/R 279, ISO/R 280, and
ISO/R 875, respectively [42].
2.2.6. Determination of citronella oil constituents
The citronella oil constituents mainly consist of geraniol,
citronellal, citronellol, citral, etc. Citronella oil constituents
in this study were identified by gas chromatography/mass spec-
573
trometric (GCMS) (Shimadzu-QP2010 Plus). GC was
equipped with a capillary column (Cross bond Carbowax,
polyethylene glycol, 30 m long, 0.25 mm ID, 0.25 mm film
thickness) that was used to identify the composition of
extracted citronella oil. The oven temperature was pro-
grammed at 60 °C for 2 min which rise to 160 °C at the rate
of 4 °C/min and holding time of 5 min. A solvent, n-hexane
(1/10 v/v) was used to dilute the samples [3] and about 8 lml
diluted samples were injected into the GC by split mode with
a ratio of 100/1. The injection temperature was set at
200 °C. The rate of helium as a gas carrier was adjusted lin-
early by 46.3 cm/sec. A calibration curve was developed using
pure citronella oil and a database from the Wiley library was
also used to compare the mass spectral fragmentation patterns
in identifying the constituents of citronella oil. The quantifying
of components was carried out concerning their compound’s
retention time. The percentage of each component was
obtained as a ratio of the area under the peak and the total
area of the chromatogram [3]. Citronella oil from the extrac-
tion of lemongrass leaves by the US-MAHD method would
be compared with citronella oil resulting from each UAE
and MAHD method.
2.2.7. Scanning Electron Microscopy analysis of lemongrass
The effect of extraction methods (UAE, MAHD, and
US-MAHD) on the structure of the lemongrass plant was
investigated by Scanning Electron Microscopy (SEM). SEM
analysis aims to investigate the ability of ultrasonic and micro-
wave energy to destruct the cell wall that facilitates the release
of oil to come out from the plant cell. The residues of lemon-
grass after the extraction process in every method were col-
lected and dried at room temperature for further SEM
analysis [3840]. The microstructures of plants including shape
and surface characters were observed and recorded by SEM
(JSM 6510 LA, JOEL Ltd, Japan). This analysis was carried
out at Laboratorium Terpadu, Universitas Diponegoro,
Semarang.
3. Results and discussion
3.1. The extraction process of citronella oil from lemongrass
leaves
The extraction of citronella oil from lemongrass leaves using
US-MAHD was carried out at various bath temperatures of
30 °C, 40 °C, and 50 °C (UAE), and microwave powers
(150 W, 300 W, and 450 W). Fig. 3 shows the yield obtained
from extraction by US-MAHD in this study. Based on obser-
vations, the overall increase in ultrasonic bath temperature
from 30 °C to 50 °C and the ratio of solvent to citronella sam-
ples from 10:1 to 14:1 decreased the yield of citronella oil. For
example, increasing the ultrasonic bath temperature at a
solvent-plant ratio of 10:1 results in a decrease in yield in the
range of 3.0 to 4.5 mg/g. As with temperature, increasing the
ratio of solvent to material also decreases yield. Increasing
the solvent-ingredient ratio from 10:1 to 14:1 decreased the
yield in the range of 3.6 to 7.3 mg/g. Meanwhile, increasing
the microwave power from 150 to 300 W increased the oil yield
up to 35.8 mg/g. However, the use of microwave power of
more than 300 W reduces the yield of citronella oil.
574 M. Sarah et al.

Fig. 3 Effect of solvent-plant ratio (v/m) on the yield of citronella oil using the US-MAHD method at various ultrasonic temperatures
and microwave power: (a) solvent-plant ratio 10:1, (b) 12:1, and (c) 14:1.

The highest yield of extraction obtained was 1.82 mg/g

Table 1 Physical properties of citronella oil.
while the lowest yield was 1.17 mg/g. The highest yield was
obtained from a combination of ratio solvent to plant of Parameters Result of this study ISO 3848:1976
10:1, the ultrasonic temperature of 30 °C, and microwave Colour Yellow-brownish Pale yellow-brownish
power of 300 W with an extraction time of 25 min in an ultra- yellow yellow
sonic bath and 65 min in a microwave oven. The lowest yield Density 0.880–0.917 0.880–0.922
of citronella oil was obtained by using a solvent to plant ratio Refractive Index 1.466–1.472 1.466–1.475
of 14:1, an ultrasonic temperature of 50 °C, and microwave Solubility in Ethanol 1:2 clear 1:2 clear
power of 150 W at a similar period. 80%
In this study, the effect of some parameters to yield citro-
nella oil such as solvent-plant ratio, the temperature of the
ultrasonic bath, and the power of the microwave oven were
tions of solvent to plant ratio, the temperature of the ultra-
investigated in the extraction process from lemongrass leaves.
sonic bath, and the power of the microwave oven. Overall
The physical properties of citronella oil extracted from lemon-
citronella oil resulting from the US-MAHD in Table 1 meets
grass leaves by using the US-MAHD method are shown in
the standard specification of ISO 3848:1976. Oil relative den-
Table 1. The physical properties indicated characteristics of
sity, refractive index, and solubility in 80% (v/v) ethanol were
citronella oil obtained from experiments at various combina-
Extraction of citronella oil from lemongrass (Cymbopogon winterianus) by sequential ultrasonic
determined following ISO/R 279, ISO/R 280, and ISO/R 875
respectively.
The colors of citronella oil produced in this study were
yellow-brownish yellow, this yellow color was from the yellow
substance contained in the essential oil called carotene [43].
The refractive index is influenced by the length of the carbon
chain and the number of double bonds. The higher the index
value, the higher the carbon chain length and the number of
double bonds. Essential oil with a large refractive index value
has better quality than oil with a small refractive index [44].
Solubility in ethanol produced a clear mixture, which meant
the oil was completely soluble in ethanol. This was following
the theory because the essential oil must be 100% soluble in
ethanol [43].
3.1.1. The effect of solvent-plant ratio of US-MAHD to yield of
the citronella oil
To determine the effect of the plant-solvent ratio on the yield
of citronella oil, an evaluation was carried out at (1) constant
ultrasonic temperature (30 °C) and (2) constant microwave
power (300 W) which gave maximum results. The solvent-
plant ratio in the extraction can affect the level of interaction
between solids and solvents which will affect the extraction
results, the best results are achieved when the solution reaches
a saturation concentration [45]. The effect of the plant-solvent
ratio on the yield of citronella oil at a constant ultrasonic bath
temperature of 30 °C is shown in Fig. 4, while Fig. 5 shows the
same effect at a constant microwave power of 300 W. Figs. 4
and 5 show the correlation between the solvent-plant ratio
and the yield of citronella oil of extraction with US-MAHD
is inversely related. Fig. 4 shows the extraction yield decreased
by 7.46 mg/g as the ratio increased from 10:1 to 14:1 at 150 W
microwave power. Similar results were obtained at 300 W and
450 W microwave power. An increase in the plant-solvent ratio
from 10: 1 to 14:1 reduces the extraction yield by 7.69 mg/g
and 8.87 mg/g at 300 W and 450 W microwave power, respec-
tively. In principle, electromagnetic waves cause the reorienta-
tion of polar water molecules according to the direction of the
magnetic field, causing friction and heat. The friction between
the water molecules and the lemongrass raw material tears the
cell walls and releases lemongrass oil. However, a substantial
Fig. 4 Effect of solvent-plant ratio (v/m) to the yield of citronella oil using the US-MAHD method at a fixed ultrasonic temperature of
30 °C and various microwave powers of 150 W, 300 W, and 450 W.
575
increase in the amount of solvent and microwave power results
in excess heat. If this happens in a long period (90 min), then
there will be evaporation of the solvent and the resulting citro-
nella oil. This resulted in a reduced oil yield as shown in Fig. 4.
Similar results were also concluded from the extraction at a
fixed microwave power of 300 W as shown in Fig. 5. Extrac-
tion of citronella oil using a microwave power of 300 W and
a temperature of 30 °C reduced the yield by 3.3 mg/g and
4.5 mg/g as the plant-solvent ratio increased from 10:1 to
12:1 and 14:1, respectively. Increasing the temperature from
30 °C to 40 °C at a constant microwave power of 300 W
decreased the yield by 9.7 mg/g (ratio of 12:1) and 2.5 mg/g
(ratio of 14:1). Increasing the temperature from 30 °C to
50 °C increases the water vapor pressure which weakens the
cavitation bubbles thereby avoiding damage to the cell walls
of the citronella plant samples.
Similar conclusions were reported by Wong et al. [46],
Ranitha et al. [3], Ho et al. [47], and Hien Tran et al. [48]. Fun-
damentally, the increment of solvent to plant ratio from 10:1
to 12:1 and 14:1 elevated the driving force of mass transfer
due to the high gradient of concentration between the solvent
and the phytochemical component. Overall increase the yield
of extraction [46]. However, in the microwave-assisted extrac-
tion process, an increase in the amount of excess polar solvent
would cause excess thermal stress [3], so some of the extracted
components will be lost. In addition, an excess amount of
polar solvent could dissolve or emulsify oil compounds so that
the extracted essential oil yield would be reduced [4748]. The
amount of solvent used for the extraction of essential oils from
plant material must be in sufficient quantities. The greater
amount of solvent can limit the overheating of the plant mate-
rial and there is no burning of the plant material. In addition, a
higher amount of solvent can also cause more energy
absorption, so the oil yield decreases [49].
3.1.2. Effect of ultrasonic temperature of US-MAHD on yield of
the citronella oil
Fig. 6 shows the effect of ultrasonic temperature on the extrac-
tion yield at a fixed microwave power of 300 W. The yield of
citronella oil decreases as the ultrasonic temperature increases.
Extraction using the US-MAHD method using a solvent to
576
Fig. 5 Effect of solvent-plant ratio (v/m) to the yield of citronella oil using the US-MAHD method at a fixed microwave power of 300 W
and various ultrasonic bath temperatures of 30 °C, 40 °C, and 50 °C.
plant ratio of 10:1 will produce a yield of 1.82 mg/g at an ultra-
sonic temperature of 30 °C. The yield decreased to 1.67 mg/g
when the ultrasonic temperature was increased to 50 °C.
Extraction yield decreased when the solvent to plant ratio
was increased to 12:1 and 14:1. When the ultrasonic tempera-
ture was increased from 30 to 50 °C, the yield decreased by
13.69 mg/g and 13.63 mg/g for the plant-solvent ratio of 12:1
and 14:1, respectively. In point 3.3.1 it has been explained that
the temperature increment of the ultrasonic bath reduces the
yield of citronella oil. An increase in temperature will increase
the water vapor pressure thereby lowering the surface tension
and reducing the effect of cavitation bubbles due to the move-
ment of water vapor into the bubbles which weakens the bub-
bles. As a result, the contact between the solvent and the plant
sample was not optimal and this minimized damage to the sur-
face of the citronella plant sample so that the oil released was
reduced.
Chemat et al. [50] reported similar results elsewhere. An
increase in ultrasonic temperature would elevate the vapor
pressure of the solution that decreasing viscosity and surface
tension. It will reduce the effect of cavitation in the extraction
process because the solvent vapor would enter the cavitation
Fig. 6 Effect of ultrasonic temperature on extraction yield at a microwave power of 300 W using the US-MAHD method.
M. Sarah et al.
bubbles and caused the bubbles to collapse weakly. It would
reduce the number of damaged cells due to the effect of cavi-
tation causing a reduction of the extracted compound. Con-
versely, selecting a low temperature such as 30 °C could
increase the yield due to the increment of cavitation bubbles
number and the solid-solvent contact area. Similar results were
also obtained on ultrasound-assisted extraction of essential oils
from Oliveria decumbens flowers, the yield of essential oils
increased significantly with increasing temperature from 25
to 40 °C. However, a further increase in ultrasonic temperature
from 40 to 55 °C caused a decrease in yield [51].
3.1.3. The effect of microwave power of US-MAHD on yield of
the citronella oil
The effect of microwave power on yield resulting from the
extraction of citronella oil using US-MAHD at a fixed ultra-
sonic temperature of 30 °C is shown in Fig. 7. Extraction using
various solvent-plant ratios increases the yield of citronella oil
as microwave power rises from 150 W to 300 W. The yield of
citronella oil increases by 35.8 mg/g, 37.5 mg/g, and 35.5 mg/g
at a solvent-plant ratio of 10:1, 12:1, and 14:1 respectively.
Increasing microwave power above 300 W decreases the yield
Extraction of citronella oil from lemongrass (Cymbopogon winterianus) by sequential ultrasonic
by 7.14 mg/g, 6.81 mg/g, and 8.33 mg/g at a solvent-plant ratio
of 10:1, 12:1, and 14:1 respectively.
Increasing the power of the microwave oven will increase
the penetration of electromagnetic waves into samples of citro-
nella plants and water solvents. This has implications for
increasing energy conversion from microwave energy into heat
energy. The mechanism of energy change from microwave
energy to heat energy has been described in point 3.3.1.
Increasing the power of the microwave oven will increase the
amount of heat energy so that excess heat occurs. Based on
observations, it is known that the microwave power of
300 W is the maximum power that can be tolerated by the
ingredients (water solvent and lemongrass plant samples) to
prevent excessive heating. Heating a microwave oven with a
power of less than 300 W produces a low yield of citronella
oil because the amount of power density produced is low,
and vice versa. The power density limits tolerated by the mate-
rial in this study were 227.72 W/kg, 230.77 W/kg, and 200 W/
kg for extraction with US-MAHD at plant-solvent ratios of
10:1, 12:1, and 14:1 respectively.
According to Hien Tran et al. [48], the use of high micro-
wave power will increase the yield of essential oils but only
to a certain power level. The increase in extraction yield
occurred because the use of high power would result in a
higher temperature rise due to the more random reorientation
of polar molecules [3]. This process could increase the solvent
diffusion so that more of the essential oil would be extracted.
However, the use of microwave power could cause some of the
extracted chemical components to be lost and decomposed
[48]. Nour et al. [52] reported that during short extraction
times, about 30 to 90 min, the yield increased with increasing
microwave power. However, when the extraction was carried
out long enough, about 120 to 150 min, the yields obtained
were similar under different microwave power. The initial
extraction rate increased with increasing microwave power.
This is due to the rapid heat generation in the extracted plant
material by absorption of microwave energy and the formation
of a higher pressure gradient in the plant material when
applied to higher microwave power.
The increase in yield at the beginning of the extraction then
decreased most likely due to the recruitment of balanced
microwave forces which disrupt the plant cell wall thereby
facilitating dissolution into the cell and mass transfer. On the
Fig. 7 Effect of microwave power on extraction yield at a fixed ultrasonic temperature of 30 °C using the US-MAHD method.
577
other hand, the target compound can be degraded by increas-
ing the microwave power, resulting in a decrease in the extrac-
tion yield. Therefore, suitable microwave power is very
important for the extraction process [36].
3.2. The comparison of UAE, MAHD, and US-MAHD methods
in the extraction process of citronella oil
A comparison of the extraction yields produced using the
UAE, MAHD, and US-MAHD methods is shown in Fig. 8.
The extraction process for each of the above methods was car-
ried out for 90 min using lemongrass leaves. Fig. 8 shows the
US-MAHD method obtained a higher yield while the lowest
yield was obtained by the UAE method. The yield of citronella
obtained by the US-MAHD method was 49.45% higher than
the UAE method, while compared to the MAHD method,
the yield obtained by the US-MAHD method increased by
18.68%.
Yu et al. [37] reported combining the UAE and MAE meth-
ods sequentially (US-MAE) results in a higher extraction yield
of flavonoids as compared to UAE or MAE methods. In the
process of extracting phytochemical compounds using the
US-MAE method, it was reported that more cell walls were
completely damaged than in the process with UAE and
MAE so that the yield of the extracted compounds could be
higher. The highest total flavonoid content was obtained in
the sequential combination of microwave-and ultrasound-
assisted extraction (SC-MUAE) method (7.85 mg/g) compared
to other methods, MAE (4,789 mg/g), UAE (6,229 mg/g), and
sequential combination of ultrasound- and microwave-assisted
extraction (SC-UMAE) (6,548 mg/g). It is related to the SEM
analysis in this study shown in Fig. 9 for extraction by UAE,
MAHD, and US-MAHD methods at 30 °C (UAE and US-
MAHD), 300 W (MAHD and US-MAHD), and 90 min.
Fig. 9(a) shows the SEM results of the sample before the
extraction process is carried out. The black color indicates that
there is no damage to the outer surface of the citronella plant
cell sample indicating that the cell wall is still intact. Fig. 9(b),
9(c), and 9(d) show the SEM of a sample after the extraction
process by UAE, MAHD, and US-MAHD respectively.
Fig. 9(b) shows the SEM of a sample with a damaged cell wall
after being treated with an ultrasonic wave to obtain citronella
578 M. Sarah et al.

Fig. 8 Yield comparison of UAE, MAHD, and US-MAHD methods in this study at 30 °C (UAE/US-MAHD), 300 W (MAHD/US-
MAHD), and 90 min.

Fig. 9 SEM Analysis of lemongrass sample pre-extraction (a), post-UAE (b), post-MAHD (c), and post-US-MAHD (d) at 30 °C (UAE/
US-MAHD), 300 W (MAHD/US-MAHD), and 90 min.
Extraction of citronella oil from lemongrass (Cymbopogon winterianus) by sequential ultrasonic 579

oil, as indicated by a red circle. A similar illustration is shown
in Fig. 9(c) and 10(d) for extraction with MAHD and US-
MAHD respectively. SEM results from extraction using the
US-MAHD method (Fig. 9(d)) showed the highest level of cell
wall damage. This can be explained as follows. Extraction by
the US-MAHD method occurs because ultrasonic waves and
microwave energy are gradually applied to the sample.
In the UAE stage, ultrasonic waves propagate through the
solvent and cause the phenomenon of acoustic cavitation
which on a micro-scale can increase the temperature and pres-
sure, thereby increasing the pressure difference and the sheer
force of the solvent. The contact between the cavitated waves
with the sample surface will break some of the sample cell walls
and the citronella oil will come out. If further extraction is car-
ried out with microwaves, the remaining solvent will be reori-
ented following the movement of polar molecules so that there
is an increase in temperature and the process of cell wall dam-
age will increase. This has implications for obtaining high
yields. This confirms why extraction with US-MAHD obtains
the highest yield. In the case of extraction using the UAE and

Fig. 10 The chromatogram of citronella oil extracted from leaves using UAE (a), MAHD (b), and US-MAHD Method (c) at 30 °C
(UAE/US-MAHD), 300 W (MAHD/US-MAHD), and 90 min.
580 M. Sarah et al.

decrease in the intensity of ultrasonic waves along with the

Table 2 Citronella oil resulted from UAE, MAHD, and US-
increase in the wave propagation distance, thus allowing the
MAHD Method.
presence of undamaged cells resulting in phytochemical com-
Compounds Relative Content (%) pounds cannot be extracted. While in the MAHD method,
UAE MAHD US-MAHD inhomogeneous heating can occur due to uneven wave absor-
Limonen 1.65 2.05 3.10
bance in the sample which will reduce the extraction yield.
Pinene 1.22 2.82 2.74 GC-MS analysis was performed to compare the com-
Carene – 1.21 0.96 pounds of citronella oil obtained by the UAE, MAHD, and
Citronella 11.52 18.03 17.79 US-MAHD methods. Chromatogram of GC-MS analysis for
Linalool 3.58 2.51 1.83 citronella oil from leaves is shown in Fig. 10 and information
Isopregol 3.14 1.66 1.98 on the comparison of compounds produced by each of the
Isopulegol – 2.81 – UAE, MAHD, and US-MAHD methods are shown in Table 2.
Citronellol Acetate 2.57 1.85 1.11 The content of chemical compounds in citronella oil obtained
Z-Citral – – 1.07 from extraction by the UAE, MAHD, or US-MAHD method
E-Citral 1.93 1.77 2.23
varies. The MAHD method produced citronella oil with the
Caryophllene 1.42 – –
Geraniol Acetate 8.12 6.24 4.44
highest citronellal content (18.03%) while the highest citronel-
Citronellol 17.25 13.00 12.45 lol (17.25%) was produced by the UAE method. The
Isoeugenol – 3.65 – US-MAHD method produced the highest geraniol (46.69%),
Geraniol 42.60 38.78 46.69 limonene (3.10%), and citral (3.30%). The difference in the
Geranyl Butyrate 2.56 2.41 1.72 content of citronella oil content from the 3 methods as shown
Alpha-terpinolene – – 1.16 in Table 2 provides a distinct advantage because it can be used
Eugenol 1.46 1.20 0.73 for various purposes such as perfume, a fragrance for house-
hold appliances, anti-fungal, anti-bacterial and antioxidant.
Each process has its advantages concerning the content of
citronella oil. This provides an opportunity for product diver-
Table 3 The compounds of citronella oil resulted from stems sification toward the use of lemongrass. US-MAHD produced
using US-MAHD method. oil with the highest content of geraniol and citral at 46.69%
The compound of Citronella Oil Relative Content (%) and 3.30%, respectively. Geraniol and citral components are
raw materials for perfumes and household fragrances. Mean-
Limonene 1.05
while, UAE produces an oil with the highest citronellol content
Origanene 1.91
Citronella 17.19
(17.25%), while MAHD produces an oil with citronellal con-
Nerol Asetate 4.41 tent (18.03%). Citronellol is a compound that is widely used
Citronellol 15.28 as an antifungal and anti-microbial agent, while citronellal
Geraniol 53.88 functions as an antioxidant.
Geranyl Butyrate 1.68 According to Wany et al. [6], the most important chemical
Isopropyl Methyl Propylidene 2.07 compounds of citronella oil were citronellal, citral, citronellol,
Dimethyl Bicyclo Hexane 2.52 and geraniol. Citronellal was a monoterpenoid that had anti-
fungal activity, and antimicrobial activity, and is widely used
as an insect repellent. Other compounds of citronella oil, citral
is one of the raw materials for producing perfume and vitamin
MAHD methods only, the cell wall destruction process is not A while citronellol contains antioxidants [53]. Maz czka et al.
as optimal as US-MAHD. The UAE and MAHD methods
[54] reported geraniol is widely used for its application as a fra-
have the disadvantage of being carried out separately. In the
grance in cosmetic and household products.
UAE method, a wave attenuation effect can occur, namely a

Fig. 11 The chromatogram of citronella oil extracted from stems using the US-MAHD Method at 30 °C, 300 W, and 90 min.
Extraction of citronella oil from lemongrass (Cymbopogon winterianus) by sequential ultrasonic
3.3. The extraction process of citronella oil from lemongrass
stems
Extraction of lemongrass stems was also conducted to com-
pare the yield and oil constituents. The process of extracting
citronella oil from lemongrass stems was carried out using
the US-MAHD method under the same conditions of leaves
extraction resulting in the highest yield. The yield resulting
from leaves and stems under this condition was 1.82 mg/g
and 0.78 mg/g respectively. The extraction yield obtained by
the stem is lower than the yield from leaves. The same result
was also reported by Iqbal et al. [55] that the essential oil
extracted from the stem of Murraya koenighi was lower as
compared to the leaves. The compounds of citronella oil from
the stem were identified using GC-MS. The result can be seen
in Table 3 and its chromatogram of GC-MS analysis is shown
in Fig. 11.
Tables 2 and 3 show the different compositions of citronella
oil extracted from the leaves (Table 2) and the stems of the
lemongrass plant (Table 3) using US-MAHD at 30 °C,
300 W, 90 min, and the ratio of the solvent to plant 1:10. It
was known that geraniol was the highest compound of citro-
nella oil (53.88%) extracted while the compound of citral
was not found. Besides that, the relative content of citronella
and citronellol extracted from stems respectively was 17.19
% and 15.28 %. The geraniol content of citronella oil
extracted from the stem (53.88%) was more than that
extracted from the leaves (46.69%). So that the stem is very
potential to be used as raw material for making perfumes.
However, the absence of the citral component and the low
lemon content (3.10%) from the citronella stems requires mix-
ing the oil from the citronella stems with the oil from the citro-
nella leaves to produce perfume and household fragrances.
Based on essential oil yield, Barbouchi et al. [56] reported that
the yield of essential oil isolated from the leaves of three Ruta
species was higher (1.86 ± 0.01%) than the yield of essential
oil isolated from the stems (0.16 ± 0.00%) (Barbouchi,
et al., 2021). Although the yield of lemongrass leaves was
2.34 times higher than the stems, lemongrass stems contain
higher citronella oil content than citronellol and geraniol.
4. Conclusions
The sequential US-MAHD method in this study was carried
out for 90 min at a combination of solvent-plant ratio of
10:1 (v/m), an ultrasonic temperature of 30 °C, and a micro-
wave power of 300 W obtained the highest yield of 1.82 mg/
g. The extracted oil has met the standard specified by ISO
3848:1976 with a characteristic odor of strong lemon scent,
color pale yellow, a relative density of 0.917 mg/l, a refractive
index of 1.472, and solubility in 80% (v/v) of ethanol was 1:2
(v/v). The oil constituents mainly consist of citronella
(17.79%), geraniol (46.69%), citronellol (12.45%), limonene
(3.10%), and citral (3.30%). Based on this study, it is con-
cluded that extraction of citronella oil from lemongrass by
US-MAHD has the following advantages: (1) Ultrasonic
extraction acts as a preliminary extraction where ultrasonic
waves damage cell walls so that some of the oil in the plant
matrix can be released. The further extraction process through
microwave heating expands the damage to the plant cell walls
due to the re-orientation of polar molecules in the citronella
581
plant which increases the temperature (volumetric heating)
and pushes more citronella oil out of the citronella plant
cell-matrix, resulting in high yield of citronella oil. (2) It does
not require too much solvent (water), so it does not require
a large extraction vessel if it is developed on a commercial
scale. (3) This extraction can be conducted with fairly low
power (300 W) so that the electrical power requirements are
not high. However, this extraction process also has several
challenges in its implementation on a commercial scale, such
as how to design inexpensive ultrasonic and microwave irradi-
ation systems. Therefore, it is necessary to develop further
research for this extraction using ultrasonic and microwave
hybrid extractor.
Declaration of Competing Interest
The authors declare that they have no known competing
financial interests or personal relationships that could have
appeared to influence the work reported in this paper.
Acknowledgments
The authors gratefully thank Universitas Sumatera Utara for
supporting this research under TALENTA Scheme for the Fis-
cal Year of 2020.
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