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The 6th International Conference on Agriculture, Environment and Food Security IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1241 (2023) 012070 doi:10.1088/1755-1315/1241/1/012070

Volatile compounds of grapefruit (Citrus Grandis (L.) Osbeck)

peel essential oil by cold pressing and hydrodistillation
methods

T H Tran1,2,*, T P Dao1,2, X T Le3,4, B L Huynh5 and L T N Minh6

1
Institute of Applied Technology and Sustainable Development, Nguyen Tat Thanh
University, Ho Chi Minh City, Vietnam.
2
Faculty of Food and Environmental Engineering, Nguyen Tat Thanh University, Ho
Chi Minh City, Vietnam.
3
Faculty of Chemical Engineering, Ho Chi Minh City University of Technology
(HCMUT), 268 Ly Thuong Kiet, District 10, Ho Chi Minh City, Vietnam.
4
Vietnam National University Ho Chi Minh City, Linh Trung Ward, Thu Duc District,
Ho Chi Minh City, Vietnam.
5
Ho Chi Minh City University of Food Industry, Ho Chi Minh City, Vietnam
6
Branch of Green Power Co., LTD, Ben Tre Province, Vietnam.

Email: *hientt@ntt.edu.vn

Abstract. The study of the essential oils (EOs) extraction with high medicinal properties is
necessary to contribute to the use of natural compounds. By using cold pressing and
hydrodistillation methods, the content of grapefruit peel (Citrus grandis (L.) Osbeck) EOs was
extracted. The chemical components of EOs was determined by GC-MS method. Results have
shown that 10-12 compounds were detected in the grapefruit peel EOs, which are mainly
monoterpenes such as D-Limonene (71.21%-74.83% and γ-Terpinene (15.70%-17.68%). The
findings from the present study demonstrated a new research direction on chemical composition
of the grapefruit EO, thereby contributing to applications ofthe natural compounds with high
medicinal properties from grapefruit.

1. Introduction
Essential oils (EOs) are aromatic molecules that are synthesized through regulatory functions in the
types of stems it creates, are a mixture of soluble organic substances, have a characteristic odor. Due to
the favorable use of natural-based products, EOs are widely applied in various fields of industries and
daily life [1-2]. One of the fruits that are highly familiar with many generations of Vietnamese people
is Citrus family. Beside its role as a seasonal fruit-bearing tree, they are also used in pharmaceuticals,
beverage, food industries and fruit consumption due to high content of organic compounds such as
essential oils, antioxidants and vitamins. Furthermore, several studies have shown that Citrus fruits are
widely applicable for the treatments of various diseases [1-3].
Grapefruit (Citrus grandis L. OsBeck) is a fruit tree which belongs to the Citrus family. Most
varieties of grapefruit contain EOs in the peel, leaves, and flowers, with the highest content (1.5-2.5%)
is present in the peel. The phytochemical constitutents in the grapefruit peel EO mainly contain D-
Limonene, α-pinene, β-mycrene, α-terpinolene, and β-pinene [4-7]. In addition, the grapefruit peel also

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The 6th International Conference on Agriculture, Environment and Food Security IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1241 (2023) 012070 doi:10.1088/1755-1315/1241/1/012070

contains many flavonoids such as naringoside, hesperidin, and diosmin which exhibit protective effects
on the capillary walls, thereby preventing complications due to broken capillaries, and indirectly
lowering blood pressure. In addition, grapefruit EO is now highly popular in pharmaceuticals and
cosmetics industries and stimulating hair growth, especially compounds with antioxidant and
antibacterial properties [8-10]. The extraction of grapefruit EO has been carried out by various methods
such as steam distillation, hydrodistillation, solvent distillation, pressing method, supercritical CO2
extraction method, and microwave-assisted distillation [2, 11-13]. Citrus EO is mainly used in
manufacture and application industries. However, the citral EO prepared by steam distillation did not
meet this requirement. Therefore, the cold pressing is often used to extract the EO. By applying
mechanical force, the aromatic oil sacs located in the outer shell of the citrus peel can be released. .
In attempts to utilize available raw materials to meet increasing consumers’ demand, it is necessary
to focus on exploiting EOs extraction method and improve oil quality. Therefore, the objective of this
study is to employ the cold pressing method to extract the EO from grapefruit peel and evaluate the
chemical composition of the EO by using chromatography coupled with mass spectrometry (GC-MS).
Therefore, the efficiency and quality of volatile components in grapefruit EO can be compared, thereby
contributing to the diversification of aromatic compounds obtained from plants.

2. Materials and methods

2.1. Grapefruits materials

Mature and healthy grapefruits (C. grandis (L.) Osbeck) with smooth skin. Approximately 20 kg of
grapefruits were selected and purchased from a local farm in Mekong Delta, Vietnam. After transport
to laboratory, the collected grapefruits were washed thoroughly with tap water to discard dirt and
impurities. The fruit peels were removed to obtain a total of 4 kg and subjected to subsequent EO
extraction.

2.2. Preparation of Grapefruit essentials oil sample

For cold-pressing method, the grapefruit peel was placed in the cold pressing device and recovered EO
by two-time pressing, as described in previous study [14]. A total of 1.4 L of liquid mixture containing
EO were collected and allowed to settle for a period of 12 hour at <10°C. Then, the mixture was
centrifuged to recover EO at 6000 rpm for 5 minutes, followed by dehydration using anhydrous Na2SO4
to obtain pure grapefruit EO.
For hydrodistillation method, a total of 150 ± 0.02 g of grapefruit grounds were transferred to each
flask after pressing and subjected to hydrodistillation method for 2 hour. Then, EO was collected and
dehydrated with anhydrous Na2SO4.

2.3. GC-MS Analysis

Before determining volatile components, EO sample (25 µL) was pre-treated by dilution in n-hexane
and dehydrated with Na2SO4 salt. The EO samples were collected and added to the analyzer, consisting
of GC Agilent 6890 N (Agilent Technologies, Santa Clara, CA, USA), column head pressure 9.3 psi,
HP5-MS column, MS 5973 under the following conditions: Hellium as carrier gas; split line 1:100;
injection volume 1.0 µL; injection temperature 250 °C; and flow rate 1.0 mL/min. The chemical
compositions of EO were compared to NIST11 library [15].

3. Results and discussion

3.1. Volatile compounds in grapefruit essential oil by cold pressed

Grapefruit EO is a volatile, dark green-colored liquid with a characteristic aroma of natural grapefruit
peel with an efficiency of 0.018%. The density of the EO is 0.86 (g/mL), which also varies according to
the chemical composition. The EO that is composed mainly of terpenic hydrocarbons has low density;

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The 6th International Conference on Agriculture, Environment and Food Security IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1241 (2023) 012070 doi:10.1088/1755-1315/1241/1/012070

whereas EO containing oxygen-containing compounds or aromatic kernals would have higher density
[16].
The chemical composition of the cold-pressed EO was determined by the GC-MS method. Table 1
and Figure 1 shows that the obtained essential oil consists of ten main components, with the predominant
components being D-Limonene (74.839%) and γ-Terpinene (15.703%) and other minor components
such as α-Phellandrene (2.393%), α-Pinene (2.351%), β-Myrcene (1.641%), β-Pinene (1.006%), α-
Bergamotene (0.547%), Sabinene (0.544) %), Terpinolene (0.540%), and α-Thujene (0.436%). D-
limonene is a clear colorless mobile liquid with a pleasant lemon-like odor that is present as a major
component of the citrus peel oil, with potential chemo-preventive and anticancer activities. According
to Njoroge et al., the volatile components of the cold pressed essential oil of grapefruit peel (C. grandis
Osbeck) from the same variety in Kenya were identified by GC and GC-MS. Results of Simon et al .
research have identified a total of 52 compounds, accounting for up to 98.8% of the EOs, with D-
limonene (94.8%), terpinene (1.8%), and pinene (0.5%) as the main compounds [17].
The main chemical constituent of the EO from C. grandis Osbeck bark was reported as D-limonene,
which accounted for 89.96±1.64%, followed by β-myrcene (4.49±0.38%), α-pinene (0.63±0.05%), 3-
carene (0.48±0.04%), and caryophyllene (0.47 ±0.04%) [18]. Deng et al. (2020) evaluated the volatile
compounds in cold-pressed grapefruit essential oil. Results have shown that the major constituent was
also D-limonene (93.33%) as the only sesquiterpene oxide, while the minor components were
numerousmonoterpenes, including sabinene (0.60), α-pinene (0.76%), β-myrcene (2.16%), and
caryophyllene oxide (0.04%) [19]. In addition, Lázaro et al. (2021) extracted grapefruit peel essential
oil by pilot-scale twin-screw method, with a maximum yield of 2.21 g (kg dry peel/min). The main
compounds of grapefruit essential oil were identified with D-limonene, accounting for 94.62% of the
total EO composition, as well as other components such as α-pinene, sabinene, myrcene, and β-
caryophyllene [20].

Table 1. Main ingredients in grapefruit essential oil by cold

pressing method
No. R.T (min) Compounds (%)
1 7.818 α-Thujene 0.436
2 8.077 α-Pinene 2.351
3 9.929 Sabinene 0.544
4 10.032 β-Pinene 1.006
5 10.904 β-Myrcene 1.641
6 11.501 α-Phellandrene 2.393
7 12.953 D-Limonene 74.839
8 14.691 γ-Terpinene 15.703
9 16.473 Terpinolene 0.540
10 31.642 α-Bergamotene 0.547

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The 6th International Conference on Agriculture, Environment and Food Security IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1241 (2023) 012070 doi:10.1088/1755-1315/1241/1/012070

Figure 1. Chromatography of grapefruit peel essential oil obtained by

cold pressing method

Figure 2. Chemical structure of some volatile components in

grapefruit essential oil

3.2. Volatile compounds in grapefruit essential oil by hydrodistillation from grapefruit grounds
The yield of the EO obtained from the distillation of pressed grounds is 0.267 mL/g, with transparent
white color and a characteristic odor of grapefruit peel.
The chemical composition of the obtained EO was determined by the GC-MS method. The Table 2
shows that the obtained EO consists of D-Limonene (71.21%) and γ-Terpinene (17.682%) as major
componentns, and other minor components such as α-Phellandrene (2.418%), β-Cymene (1.951%), α-
Pinene (1.559%), β-Myrcene (1.425%), α-Terpineol (0.986%), Terpinolene (0.951) %), β-Pinene
(0.773%), α-Bergamotene (0.425%), Terpinen-4-ol (0.369%), and α-Thujene (0.250%). The content of
grapefruit EO extraction by the pressing method was much higher than the hydrodistillation method,
with 14.25% and 0.29%, respectively. Besides, D-limonene as the main component of grapefruit EO
obtained from cold pressing and hydrodistillation accounted for 32.63% and55.74%, respectively. The

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The 6th International Conference on Agriculture, Environment and Food Security IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1241 (2023) 012070 doi:10.1088/1755-1315/1241/1/012070

grapefruit EO in the study contained high levels of nitrosative and oxidizing compounds that can be
associated with enhanced sleep quality and reduced cardiovascular disease [3]. Differences in the
chemical composition and quality of EOs depend on many factors such as genetics, variety, soil,
weather, and extraction conditions or method of use. Typically, the D-limonene content of
hydrodistillation EO is lower than cold-pressing method, indicating high quality of the extracted
grapefruit EO.

Table 2. Main ingredients in grapefruit essential oil by hydrodistillation method

No. R.T (min) Compounds (%)
1 7.819 α-Thujene 0.250
2 8.078 α-Pinene 1.559
3 10.034 β-Pinene 0.773
4 10.906 β-Myrcene 1.425
5 11.504 α-Phellandrene 2.418
6 12.658 β-Cymene 1.951
7 12.992 D-Limonene 71.210
8 14.711 γ-Terpinene 17.682
9 16.474 Terpinolene 0.951
10 21.238 Terpinen-4-ol 0.369
11 21.854 α-Terpineol 0.986
12 31.643 α-Bergamotene 0.425

Figure 3. Chromatography of grapefruit peel essential oil obtained by

hydrodistillation method

4. Conclusions
Grapefruit EO was successfully by using cold pressing and hydrodistillation methods yields of 0.018%
and 0.267 mL/g, respectively. EO obtained from pressing method has dark green color with scent
characteristic of fresh grapefruit, while hydrodistillation method produces a transparent oil liquid, with
a light aroma, water insolubility and volatility. The produced EO were using GC-MS analysis method.

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The 6th International Conference on Agriculture, Environment and Food Security IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1241 (2023) 012070 doi:10.1088/1755-1315/1241/1/012070

The main components of EO included D-Limonene, γ-Terpinene, β-myrcene, α-phellandrene, and α-

pinene. Therefore, it is necessary to investigate the variations in the EO composition under different
environmental conditions and distillation methods, thereby expanding the research on applicability of
the EO components and developing suitable extraction methods to effectively recover the highly
applicable components

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The 6th International Conference on Agriculture, Environment and Food Security IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1241 (2023) 012070 doi:10.1088/1755-1315/1241/1/012070

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Acknowledgements
This research is funded by Nguyen Tat Thanh University, Ho Chi Minh City, Vietnam; Science and
Technology Department of Ben Tre Province, Vietnam; Ministry of Science and Technology of Vietnam
(Project No. 50/2020.NTMN.TW, 05/08/2020).