Natural Product Research

Formerly Natural Product Letters

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Sesquiterpenes rich essential oil from Garcinia

celebica L. and its cytotoxic and antimicrobial
activities

Wen-Nee Tan, Zi-Hui Tan, Nurul Izzati Zulkifli, Nik Nur Syazni Nik Mohamed
Kamal, Nur Amiera Syuhada Rozman, Woei-Yenn Tong, Chean-Ring Leong &
Jun-Wei Lim

To cite this article: Wen-Nee Tan, Zi-Hui Tan, Nurul Izzati Zulkifli, Nik Nur Syazni Nik Mohamed
Kamal, Nur Amiera Syuhada Rozman, Woei-Yenn Tong, Chean-Ring Leong & Jun-Wei Lim (2019):
Sesquiterpenes rich essential oil from Garcinia�celebica L. and its cytotoxic and antimicrobial
activities, Natural Product Research, DOI: 10.1080/14786419.2019.1569012

To link to this article: https://doi.org/10.1080/14786419.2019.1569012

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NATURAL PRODUCT RESEARCH
https://doi.org/10.1080/14786419.2019.1569012

SHORT COMMUNICATION

Sesquiterpenes rich essential oil from Garcinia celebica

L. and its cytotoxic and antimicrobial activities
Wen-Nee Tana , Zi-Hui Tanb, Nurul Izzati Zulkiflic,
Nik Nur Syazni Nik Mohamed Kamalc, Nur Amiera Syuhada Rozmand,
Woei-Yenn Tongd, Chean-Ring Leongd and Jun-Wei Lime
a
School of Distance Education, Universiti Sains Malaysia, Penang, Malaysia; bSchool of Chemical
Sciences, Universiti Sains Malaysia, Penang, Malaysia; cCluster of Integrative Medicine, Advanced
Medical and Dental Institute, Universiti Sains Malaysia, Penang, Malaysia; dUniversiti Kuala Lumpur,
Malaysian Institute of Chemical and Bioengineering Technology, Melaka, Malaysia; eDepartment of
Fundamental and Applied Sciences, Universiti Teknologi PETRONAS, Perak, Malaysia

ABSTRACT ARTICLE HISTORY

Garcinia celebica L., locally known as “manggis hutan” in Received 30 November 2018
Malaysia is widely used in folkloric medicine to treat various Accepted 3 January 2019
diseases. The present study was aimed to examine the
chemical composition of the essential oil from the leaves of G. KEYWORDS
celebica L. (EO-GC) and its cytotoxic and antimicrobial potential. Garcinia celebica L.;
essential oil; sesquiterpenes;
EO-GC obtained by hydrodistillation was analysed using capil- MCF-7; antimicrobial
lary GC and GC-MS. Twenty-two compounds were identified,
dominated by a-copaene (61.25%), germacrene D (6.72%) and
b-caryophyllene (5.85%). In the in vitro MTT assay, EO-GC exhib-
ited significant anti-proliferative effects towards MCF-7 human
breast cancer cells with IC50 value of 45.2 lg/mL. Regarding the
antimicrobial activity, it showed better inhibitory effects on
Gram-positive bacteria than Gram-negative bacteria and none
on the fungi and yeasts tested.

CONTACT Wen-Nee Tan tanwn@usm.my

Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2019.1569012
ß 2019 Informa UK Limited, trading as Taylor & Francis Group
2 W.-N. TAN ET AL.

1. Introduction
Essential oils (EOs) are volatile substances of plants, comprised of a complex mixture
of terpenes and their oxygenated derivatives, alcohols, ethers, aldehydes, ketones and
phenols (Sharifi-Rad et al. 2017). Studies have shown that EOs and their individual con-
stituents were found to exhibit various biological activities (Sharifi-Rad et al. 2017).
Garcinia celebica L., locally known as “manggis hutan” in Malaysia, is a small to
medium-sized fruit tree native to Southeast Asia. The tree can grow up to 6 m, but
some can reach 30 m tall. It has a straight trunk and is densely branched, bearing
edible fruits which resemble small mangosteen (Lim 2012). In folkloric medicine, the
decoction of the root is given to a woman after childbirth against infection. It was
recorded that the leaves have been used by the Malays in the treatment of itches in
Peninsular Malaysia (Burkill 1966; Lim 2012).
Previous studies have shown that EOs from the genus of Garcinia exhibited various
biological activities. Okhale et al. (2016) reported that the leaf oil of G. kola dominated
by citronellic acid (48.3%) showed antimicrobial activity against S. aureus, E. coli, K.
pneumonia, C. albicans, P. aeruginosa and M. bovis BCG. In a study conducted by Tan
et al. (2018), the leaf oil of G. atroviridis which consisted mainly b-farnesene, was found
to show potent anti-proliferative effect on MCF-7 cancer cells while stimulating the
growth of BEAS-2B normal cells. Considering the traditional uses of G. celebica L. as
medicinal agents, the present study examined the chemical composition of EO-GC and
its cytotoxic and antimicrobial activities to evaluate its possible use as an alternative
agent in pharmaceuticals. To the best of our knowledge, this is the first report on the
chemical composition and biological activities of EO-GC from Malaysia.

2. Results and discussion

2.1. Compositions of essential oil
EO-GC possessed an earthy herbal aroma complimented by a tinge of sweetness.
Table S1 lists the chemical constituents identified, experimental and literature reten-
tion indices, together with their GC-FID peak area % on HP-5 column. Twenty-two
compounds, comprising 99.75% of the total oil by peak area % were identified.
Sesquiterpenes were among the main constituents detected. Among them, a-copaene
was the most dominant constituent, accounting for 61.25% of the sample, followed by
germacrene D (6.72%) and b-caryophyllene (5.85%). a-Copaene, a major constituent
found in the EO-GC, was reported to possessed a woody, spicy and honey odour,
which probably contributes to part of the earthy herbal scent of EO-GC (Cozzolino
et al. 2016). Several studies had revealed that a-copaene containing oils exhibited anti-
genotoxic, antioxidant, anticarcinogenic, hepatoprotective, anti-inflammatory and anti-
microbial properties (Turkez et al. 2014; Martins et al. 2015). b-Caryophyllene, an
ubiquitous sesquiterpene was found at a lower amount in the EO-GC (5.85%) com-
pared to other Garcinia species, such as in G. imberti (38.1%), G. rubro-echinata (37.
9%), G. talbotii (30.4) and G. mangostana (21.1%). It is primarily known for woody and
spicy odour and has been generally used as a fragrance additive in spice blends,
soaps, detergents, cosmetics, food products and beverages (Hartsel et al. 2016).
 NATURAL PRODUCT RESEARCH 3

Previous findings have proven that this sesquiterpene possessed antimicrobial, anti-
inflammatory, anticarcinogenic, antibiotic, antioxidant and local anaesthetic effects
(Legault and Pichette 2007; Huang et al. 2012).

2.2. Cytotoxic activity

Various studies have shown that EOs and their constituents could be active against vari-
ous cancer cells (Bayala et al. 2014). In this study, the in vitro cytotoxic activity of EO-GC
against MCF-7 human breast cancer cells was examined for the first time. The growth-
inhibitory effects of EO-GC were determined using an MTT assay after 24 to 72 h of
treatment. The results are illustrated in Figure S1. Based on the figure, the increased
concentration of EO-GC adversely decreased the proliferation of MCF-7 cells, with a
highly significant effect (p < 0.001). Moreover, EO-GC at concentration of 50, 70 and
100 mg/mL demonstrated a significant decrease in viability of MCF-7 cells to 33.1 ± 0.1%,
5.7 ± 0.1% and 2.8 ± 0.3%, respectively, after 24 h of treatment. Furthermore, the
obtained results also revealed that the proliferation of MCF-7 cells was consistently
inhibited by EO-GH for up to 72 h of incubation period. The highest applied concentra-
tion (100 mg/mL) showed the highest decrease in cell proliferation of MCF-7 cells, where
the viability recorded 2.8 ± 0.3%, 2.8 ± 0.9% and 3.0 ± 0.2% after 24, 48 and 72 h of treat-
ment, respectively. According to the study conducted by Wright et al. (2007), 100% of
MCF-7 cell kills were observed for individual single components, a-copaene and b-car-
yophyllene at 100 mg/mL. In the same study, it was reported that the 1:1 binary mixture
of these components exhibited a notable percentage cell kills (71%) against MCF-7 at
50 lg/mL. Based on our results, synergistic effects between the constituents in the EO-
GC may take place which enhanced the cytotoxic effects against MCF-7 cells. In agree-
ment with this was the observation of the cell viability in the EO-GC, which significantly
reduced to 3% after 72 h post treatment. Thus, it is vital to take into account of all the
other minor constituents present in the EOs as they might exhibit greater bioactivity
when they are combined together (Bhalla et al., 2013; Blowman et al. 2018).
In this study, the constant IC50 values of EO-GH was graphically established based
on the dose-response curves obtained from each incubation period. The IC50 value
obtained is shown in Figure S2. The curve was constantly decreased against the treat-
ment duration, which eventually reached a constant effect and produced a constant
IC50 value of 45.2 lg/mL. In comparison to the leaf oil of G. atroviridis, which showed
an IC50 value of 71 lg/mL, the present findings reveal the potential of this plant
towards breast cancer treatment with lower IC50 value (Tan et al. 2018).

2.3. Antimicrobial activity

The EO-GC exhibited antimicrobial activity on 2 Gram-positive bacteria (B. subtilis and
MRSA) and 1 Gram-negative bacteria (P. mirabilis) (Table S2). In general, EOs are more
susceptible to Gram-positive bacteria than Gram-negative bacteria (Trombetta et al.
2005). Gram-negative bacteria, which possess a peptidoglycan layer covered by an
outer membrane, render them resistant to certain natural extracts or compounds
(Nazzaro et al. 2013). The present results are in accordance to those of previous
4 W.-N. TAN ET AL.

studies on EOs from Garcinia, indicating that Gram-negative bacteria are more resist-
ant to EOs than Gram-positive bacteria (Tan et al. 2013). Based on the results, no
inhibition was shown on yeasts and fungi tested at 10 mg/mL. This could due to the
production or secretion of certain enzymes by the fungi or yeasts which detoxify the
EO constituents into inactive forms (Stupar et al. 2014).
The MIC of EO-GC was ranged from 1.25 to 2.5 mg/mL (Table S3). Based on the
findings, the MLC was significantly higher than MIC as a higher concentration of EO
was needed to kill the microorganisms. EOs representing a complex mixture of differ-
ent chemical constituents and can be divided into hydrocarbons and oxygenated com-
pounds. Previous studies had shown that terpenoids, classified under oxygenated
compounds, exhibited significant antimicrobial activity than the other constituents
(Zengin and Baysal 2014). In this study, EO-GC was found to constitute mainly non-
oxygenated sesquiterpenes (95.61%), thus, suggesting the reduction of antimicrobial
activity evaluated in the present study.

3. Conclusions
EOs are a rich source of biological and chemical diversity. The chemical identification
and biological evaluations of EO-GC fills the knowledge gap in the genus of Garcinia.
The EO-GC analysed by capillary GC and GC-MS was found to dominated by
a-copaene (61.25%), followed by germacrene D (6.72%) and b-caryophyllene (5.85%).
The cytotoxic assay showed that EO-GC exhibited significant anti-proliferative effect
towards MCF-7 human breast cancer cells. In the antimicrobial study, it showed anti-
bacterial effects against B. subtilis, MRSA and P. mirabilis, with MIC values ranged from
1.25 to 2.5 mg/mL. Further investigations are warranted to employ EO-GC as potential
chemotherapeutic agent in breast cancer treatment.

Disclosure statement
No potential conflict of interest was reported by the authors.

Funding
This work was supported by the USM Short Term Grant (304.PJJAUH.6313235) of Universiti Sains
Malaysia, Malaysia.

ORCID
Wen-Nee Tan http://orcid.org/0000-0002-7812-3946
Jun-Wei Lim http://orcid.org/0000-0003-0158-8822

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