Identification of Potential Antioxidants from Leaves
of Eucalyptus grandis PT Toba Pulp Lestari, Tbk.
Rizky Hidayati
1
, Muhammad Taufik
2*
, Zul Alfian
2
, Sovia Lenny
2
, Chintya Cahaya
1
,
Simon Sidabuke
3
and E. Manullang
3
1
Postgraduate Chemistry Study Program, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara,
Jl. Bioteknologi No. 1 Kampus USU, Medan, Indonesia
2
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Jl. Bioteknologi
No. 1, Medan 20155, Indonesia
3
PT Toba Pulp Lestari, Tbk., Indonesia
Keywords: Eucalyptus, Antioxidants, DPPH Method, GCMS, grandis.
Abstract: Eucalyptus plants are one type of essential oil that is fast growing and is also known as a plant that can
survive in the dry season and has a deep root system. This essential oil is widely used in various fields, such
as for pharmaceuticals, cosmetics and the food industry both as antioxidants and antibacterial. This study
aims to identification of potential antioxidants and the main active compounds from leaves of Eucalyptus
grandis PT. Toba Pulp Lestari, Tbk. Extraction was carried out using solvents, methanol, ethanol and
dichloromethane. The antioxidant potential test was determined by the DPPH method (1,1 diphenyl
picrylhydrazyl). The variation of sample volume 20; 30; 40; 50; 60 μL. Antioxidant activity with IC50
Eucalyptus grandis leaf extract was 5,349 μg / mL. Eucalyptus leaf extract is categorized as providing weak
antioxidant activity. The main active compound found is Sineol. The peak area was observed at 52% at
retention time 13.605. The other compounds obtained in this leaf were α pinene, β pinene, 1,3,7-Octatriene,
3-Ethylpentane, Paracymene, and terpinene.
1 INTRODUCTION
Eucalyptus leaves can produce essential oils or better
known in trade as eucalyptus oil. Essential oils or
essential oils are produced by aromatic plants
originating from shoots, flowers, leaves, stems, seeds,
fruits, roots, wood, and bark (Teixeiraet al., 2012).
Essential oils are natural compounds that are volatile
and very complex with strong odors. Essential oils
have bactericidal, fungicidal and insecticidal
properties (Filiptsovaet al., 2017). There are more
than 60 compounds contained in essential oils
(Bakkali et al., 2008). Eucalyptus plants (Myrtaceae)
have various species, such as E. camadulensis, E.
grandis, E. pellita, E. tereticornis, and E. Torreliana.
Eucalyptus plants are fast growing plants, have
many benefits, both in terms of stem, branches, and
leaves. PT Toba Pulp Lestari Tbk has developed
eucalyptus plants to be used as paper raw materials.
However, what is still used from Eucalyptus plants
is still in the wood and branches, while the leaves
have not been processed further.
The eucalyptus species is an aromatic plant that
has antimicrobial and antioxidant properties from
the essential oils produced. The essential oils
produced are used in pharmaceutical and cosmetic
products (Ait-Ouazzou et al., 2011; Santos et al.,
2011). Eucalyptus leaves can be processed with
several extraction techniques, such as hydro-
distillation and steam extraction (Zhaoet al., 2014;
Singh et al., 2016). Analysis of the composition of
essential oils can be analyzed by the gas
chromatography method (Burt, 2004).
Previous researcher (Abdul-Majeed, 2013)
University of Baghdad carried out distillation using
the Stahl Tool, proving that eucalyptus oil contained
large levels of cineol which reached 72.71%. By
conducting research on the same species, (Cheng,
2008) distilled the Stahl Tool on Eucalytus
europhylla plants obtained 58.34% cineol levels
which was far more than steam distillation.
Hidayati, R., Taufik, M., Alfian, Z., Lenny, S., Cahaya, C., Sidabuke, S. and Manullang, E.
Identification of Potential Antioxidants from Leaves of Eucalyptus grandis PT Toba Pulp Lestari, Tbk..
DOI: 10.5220/0008920202530256
In Proceedings of the 1st International Conference on Chemical Science and Technology Innovation (ICOCSTI 2019), pages 253-256
ISBN: 978-989-758-415-2
Copyright
c
 2020 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved
253

Lack of utilization of eucalyptus leaves in the
area of PT. Toba Pulp Lestari which has the
potential to be used as raw material for making
essential oils or better known as tradable eucalyptus
oil, it is necessary to do research on the composition,
antioxidant potential, and cytotoxic activity on
Eucalyptus grandis leaves. The main oil
composition that determines the quality of quality is
based on the levels of cineol contained in it. (Copper
et al., 1991). The greater the content of cineol in oil,
the better the quality of eucalyptus oil produced.
2 MATERIALS AND METHODS
2.1 Materials
The main material for this research is Eucalyptus
grandis. The solvent used in the distillation process
is aquadest. The reagents used in the DPPH test are
2,2-diphenyl-1-pikrilhydrazil (DPPH) and methanol.
The instrumentation used for sampling samples
included the 1800 Shimadzu UV-Vis spectrometer.
2.2 Preparation of Sample
The fresh leaves obtained are immediately separated
from the stem. Then the leaves are chopped using a
cutter and scissors to produce chopped ingredients
with a length of ± 0.5-1.0 cm.
Plant identification has been carried out in the
HERBARIUM MEDANENSE (MEDA) Laboratory
in the Department of Biology, University of North
Sumatra. Samples in the form of: Fresh leaves in a
single branch between 10-15 cm from the top.
2.3 Sample Extraction
Eucalyptus leaves dried for 24 hours at room
temperature. Then the sample is weighed as much as
150 grams and inserted into a 1000 mL size flask.
Adding aquabides to taste is then connected to a
Stahl distiller, and boiled for ± 5-6 hours at ± 100 °
C to produce oil and distillation ends when the
distillate is clear. The essential oil obtained is
accommodated in the Erlenmeyer glass. The
distillate obtained is a mixture of oil and water. Then
the oil layer was added to CaCl
2
anhydrous to bind
water which might still be mixed with essential oils,
the oil layer was decanted and put into vial bottles,
stored in a coolant in a bottle and tightly closed.
Then extracted samples are stored in glass bottles for
further analysis.
2.4 Characterization
2.4.1 Analysis of GC-MS
The Specifications Instrument GC-MS QP 2010S
Shimadzu, using Column 5MS with type of ion
source Electron Impact, Injector Temperature:
300
0
C, Carrier Temperature: 50
0
C, Carrier: Helium,
Gas flow rate of carrier: 1.0 mL / min, Temperature
oven: 50
0
C for 5 minutes then 240
0
C for 7 minutes.,
Ionization electron: 70 ev.
The solution of each 1 μL standard sineol series
was inserted into the syringer to be injected into the
GCMS. Only the conditions adjusted to the
conditions of each piece of equipment and then
observed Mass Chromatogram data generated
interpreted data. Obtained data then in Perform
calculations to get the calibration curve and do the
determination of levels through the equation.
2.4.2 Analysis of Antioxidant Potential
Analysis of Antioxidant Potential in Essential Oils
with Ultraviolet-Visible Spectrophotometry Method
(UV-Vis).
3 RESULTS AND DISCUSSION
Antioxidant test of eucalyptus oil used DPPH
method. The results can be seen in Table 1. The
antioxidant activity test showed that eucalyptus oil
produced in this study showed IC50 value = 5.349.
This results shows that the eucalyptus oil produced
has a weak antioxidant value.
Table 1: Antioxidant test of Eucalyptus grandis oil.
Volume
sample
(µL)
Repetition
A
DPPH
A
sample
I (%)
20
1
0.643
0.303
52.87
2
0.303
52.87
3
0.303
52.87
30
1
0.216
66.40
2
0.216
66.40
3
0.216
66.40
40
1
0.232
63.91
2
0.232
63.91
3
0.232
63.91
50
1
0.163
74.65
2
0.163
74.65
3
0.163
74.65
60
1
0.159
75.27
2
0.159
75.27
3
0.158
75.42
ICOCSTI 2019 - International Conference on Chemical Science and Technology Innovation
254

Table 1 shows that volume of sample was used 20
until 60 μl. The repeat the experiment carried out 3
times. The wavelength of DPPH was 0,643. The
absorbant value of sample decreases with increasing
sample volume (0.303 – 0.158) This result is in
accordance with the results obtained by molyneux
(2004) . This causes that % inhibition to increase. By
using the IC50 calculation formula, an IC50 value of
5,349 was obtained. Based on the IC50 values
obtained indicate that the eucalyptus oil produced
has a weak antioxidant value (Ait-Ouazzou, 2011) .
DPPH is a purple organic nitrogen radical that is
purple. The presence of a radical-reducing
compound will reduce DPPH radicals by donating
hydrogen atoms to form diphenyl picrilhidrazine
(non-radical) compounds which can be characterized
by changes from DPPH radical purple to yellow
(picril group) (Molyneux, 2004). This process can be
seen in Figure 1.
Figure 1: DPPH reaction.
The analysis of GCMS (Figure 2) shown of the
sineol compound content in the peak area of 52%
and a retention time of 13,605.
Figure 2: The Chromatogram of Eucalyptus oil.
Figure 2 shown that the main active compound
found is Sineol. The peak area was observed at 52%
at retention time 13.605. The other compounds
obtained in this leaf were α pinene, β pinene, 1,3,7-
Octatriene, 3-Ethylpentane, Paracymene, and
terpinene.
4 CONCLUSIONS
Eucalyptus oil as a result of E. grandis leaves was
carried out by antioxidant testing using the DPPH
method (1.1 diphenyl picrylhydrazyl). Antioxidant
test results showed that E. grandis oil had weak
antioxidant activity. Sineol is the main active
compound in Eucalyptus oil (the peak area 52% and
RT 13.605). The other compounds obtained in this
leaf were α pinene, β pinene, 1,3,7-Octatriene, 3-
Ethylpentane, Paracymene, and terpinene.
ACKNOWLEDGEMENTS
This research was funded by the DRPM Republic of
Indonesia Ministry of Research and Technology
Republic of Indonesia Fiscal Year 2019.
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